ES2359395T3 - AMIDAS OF ANTRANILIC ACID AND ITS PHARMACEUTICAL USE. - Google Patents

Abstract

An anthranilic acid amide of formula I, ** Formula ** wherein Ar is represented by the N-oxide of subformula Ib, and R1 represents trifluoromethyl, and R2 represents bromine, propyl, propenyl or propynyl, or a tautomer thereof , or a salt of said anthranilic acid amide or its tautomer.

Description

The invention relates to new anthranilic acid amide derivatives, processes for the preparation of these, the application of these in a process for the treatment of the human or animal organism, the use thereof alone or in combination with one or the other compound. pharmaceutically active - for the treatment especially of a neoplastic disease, such as a tumor disease, of retinopathy and age-related macular degeneration; and the use of said compound - alone or in combination with one or another pharmaceutically active compound - for the manufacture of a pharmaceutical preparation (medicament) for the treatment of a neoplastic disease, retinopathy or age-related macular degeneration.

Certain diseases are known to be associated with deregulated angiogenesis, for example diseases caused by ocular neovascularization, such as retinopathies (including diabetic retinopathy), age-related macular degeneration, psoriasis, hemangioblastoma, hemangioma, arteriosclerosis, inflammatory diseases, such as rheumatoid or rheumatic inflammatory diseases, especially arthritis, such as rheumatoid arthritis, or other chronic inflammatory disorders, such as chronic asthma, arterial or post-transplant atherosclerosis, endometriosis, and especially neoplastic diseases, for example the so-called solid tumors and liquid tumors ( such as leukemia).

At the center of the network of growth regulation and differentiation of the vascular system and its components during embryonic development, normal growth and in a large number of diseases and pathological abnormalities, is the angiogenic factor known as "Vascular Endothelial Growth Factor" (VGEF), a dimeric disulfide-linked 46-KDa glycoprotein, along with its cellular receptors (see Breier, G., et al., Trends in Cell Biology 6, 454-6 [1996]).

VEGF receptors are receptor tyrosine kinases of the transmembrane type. Several types of the VEGF receptor are known, for example VEGFR-1, VEGFR-2, and VEGFR-3.

A large number of human tumors, especially gliomas and carcinomas, express high levels of VEGS and its receptors. This has led to the hypothesis that VEGF released by tumor cells could stimulate the growth of blood stacks and proliferation of the tumor endothelium in a paracrine manner and, therefore, through improved blood delivery, accelerate the tumor growth Direct evidence of the role of VEGF as a tumor angiogenesis factor in vivo has been obtained from studies in which VEGF activity was inhibited by antibodies.

Angiogenesis is considered as an absolute prerequisite for those tumors that grow beyond a maximum diameter of approximately 1-2 mm; Up to this limit, oxygen and nutrients can be supplied to tumor cells by diffusion.

The three main mechanisms play an important part in the activity of angiogenesis inhibitors against tumors: 1) inhibition of the growth of vessels, especially capillaries, in vascular resting tumors, with the result that there is no net tumor growth due to balance achieved between apoptosis and proliferation; 2) prevention of the migration of tumor cells due to the absence of blood flow to and from tumors; and 3) inhibition of endothelial cell proliferation, thus avoiding the stimulating effect of paracrine growth on the surrounding tissue exerted by endothelial cells, which normally line the vessels.

In WO00 / 27820, compounds belonging to the anthranilic acid amide class are described, compounds that are reported to inhibit the tyrosine kinase activity of the VEGF receptor, tumor growth and VEGF-dependent cell proliferation.

Surprisingly, it has now been found that the anthranilic acid amide derivatives according to the invention, described below, have advantageous pharmacological properties and inhibit, for example, the activity of the tyrosine kinase of the VEGF receptor, of tumor growth and of VEGF-dependent cell proliferation.

The anthranilic acid amide derivatives according to the invention are suitable, for example, to be used in the treatment of diseases, especially for the diseases in the treatment and also for the prevention of which, an inhibition of angiogenesis and VEGF receptor tyrosine kinase shows beneficial effects.

The invention relates to amides of the anthranilic acid of formula I,

image 1

where Ar is represented by the N-oxide of sub-formula Ib,

image 1

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R1 represents trifuoromethyl, and

R2 represents bromine, propyl, propenyl, or propynyl, or

a tautomer of these,

10 or a salt of said anthranilic acid amide, or its tautomer.

The general terms used above and hereafter preferably have the following meanings within the context of this disclosure, unless otherwise indicated:

The "lower" prefix indicates a radical having up to and including a maximum of 7, especially up to and including a maximum of 4 carbon atoms, the radicals in question being both linear and branched with one or

15 various ramifications,

Where the plural form is used for compounds, salts, and the like, it is also understood as a single compound, salt, or the like.

Any of the asymmetric carbon atoms (for example in compounds of formula I, wherein R 9 is a lower alkyl), may be present in the configuration (R) -, (S) - or (R, S) -, preferably in the configuration (R)

20 or (S) -. The compounds may also be present as mixtures of isomers or as pure isomers, preferably as pure enantiomer diastereomers.

The invention also relates to possible tautomers of the compounds according to the invention.

In the preferred embodiment, the alkyl has up to a maximum of 12 carbon atoms and is especially a lower alkyl.

The lower alkyl is preferably an alkyl from and including 1, to and including 7, preferably from and including 1 to and including 4, and is linear or branched; preferably, the lower alkyl is butyl, such as n-butyl, sec-butyl, isobutyl, terbutyl, propyl, such as n-propyl or isopropyl, ethyl or preferably methyl.

The term "perfluoro lower alkyl", as used herein means a lower alkyl radical wherein all hydrogen atoms are replaced by fluorine atoms.

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Halogen is especially fluorine, chlorine, bromine, or iodine, especially fluorine, chlorine, or bromine.

Such salts are formed, for example, as acid addition salts, preferably with organic or inorganic acids, from the compounds according to the invention with a basic nitrogen atom, especially pharmaceutically acceptable salts. Appropriate inorganic acids are, for example, halogenated acids, such as hydrochloric acid, sulfuric acid, or phosphoric acid. Suitable organic acids are, for example, carboxylic, phosphonic, sulfonic or sulfamic acids, for example acetic acid, propionic acid, octanoic acid, decanoic acid, dodecanoic acid, glycolic acid, lactic acid, fumaric acid, succinic acid, adipic acid, pimelic acid, subic acid, azelaic acid, malic acid, tartaric acid, citric acid, amino acids, such as glutamic acid or aspartic acid, maleic acid, hydroximaleic acid, methylmaleic acid, cyclohexanecarboxylic acid, adamantanecarboxylic acid, benzoic acid, salicylic acid, acid 4-aminosalicylic, phthalic acid, phenylacetic acid, mandelic acid, cinnamic acid, methane-or ethanesulfonic acid, 2-hydroxyethanesulfonic acid, ethane-1,2-disulfonic acid, benzenesulfonic acid, 2-naphthalenesulfonic acid, 1,5naphthalenedisulfonic acid, acid 2-, 3-or 4-methylbenzenesulfonic acid, methylsulfuric acid, ethyl acid sulfuric acid, dodecyl sulfuric acid, N-cyclohexylsulfamic acid, N-methyl-, N-ethyl- or N-propyl-sulfamic acid, or other organic protonic acids, such as ascorbic acid.

For the purposes of isolation or purification, it is also possible to use pharmaceutically unacceptable salts, for example picrates or perchlorates. For therapeutic use, only pharmaceutically acceptable salts or free compounds are employed (where appropriate in the form of pharmaceutical preparations), and these are, therefore, preferred.

In view of the close relationship between the novel compounds in the free form and those in the form of their salts, including those salts that can be used as intermediates, for example in the purification or identification of the novel compounds, any reference to the compounds Free above and from now on it should be understood that it also refers to the corresponding salts, as appropriate and timely.

The compounds according to the invention have valuable pharmacological properties, as described and hereafter.

The efficacy of the compounds of the invention as inhibitors of the tyrosine kinase activity of the VEGF receptor can be demonstrated as follows:

VEGF receptor tyrosine kinase activity test. The test is carried out using Flt-1 tyrosine kinase of the VEGF receptor. The detailed procedure is as follows: 30 µl of kinase solution (10 ng of the Flt-1 kinase domain, Shibuya et al., Oncogenes 5, 519-24 [1990]) in Tris • 20 mM HCl pH 7, 5, 3 mM manganese dichloride (MnCl2), 3 mM magnesium chloride (MgCl2), 10 µM sodium vanadate, 0.25 mg / ml polyethylene glycol (PEG) 20000, 1mM dithiothreitol and 3 µg / µl poly (Glu, Tyr ) 4: 1 (Sigma, Buchs, Switzerland), 8 µM [33P] ATP (0.2 µCi), 1% dimethyl sulfoxide, and 0 to 100 µM of the compound being tested, incubated together for 10 minutes at room temperature. The reaction is then terminated by the addition of 10 µl of ethylenediaminetetraacetate (EDTA) 0.25 M pH 7. Using a multichannel dispenser (LAB SYSTEMS, USA), a 20 µl aliquot is applied to a PVDF membrane (= polyvinyl difluoride) Immobilon P (Millipore, USA), through a Millipore microtiter manifold filter and connected in vacuo. After complete removal of the liquid, the membrane is washed 4 times successively in a bath containing 0.5% phosphoric acid (H3PO4) and once with ethanol, incubated for 10 minutes each time while stirring, then mounted on a Hewlett Packard's Manifold TopCount and radioactivity was measured after the addition of 10 µl of Microacint® (liquid β-scintillation counter). The IC50 values were determined by a linear regression analysis of the percentages for the inhibition of each compound in three concentrations (as a rule 0.01, 0.1, and 1 µmol). The IC50 values that can be found with the compounds according to the invention are in the range of 0.001 to 1 µM, preferably in the range of 0.001 to

0.1 µM.

The antitumor efficacy of the compounds of the invention can be demonstrated in vivo as follows:

In vivo activity in the immunologically deficient mouse xenotransplant model: immunologically deficient female mice BALB / c (8-12 weeks of age), Novartis Animal Farm, Sisseln, Switzerland) were kept under sterile conditions with water and food at will. Tumors are induced either by subcutaneous injection of tumor cells in mice (eg, Du 145 prostate carcinoma cell line (ATCC No. HTB 81; see Cancer Research 37, 4049-58 (1978)) or by implantation of tumor fragments (approximately 25 mg) subcutaneously on the left flank of mice using a 13 gauge trocar syringe under Foreno® anesthesia (Abbott, Switzerland) Treatment with the test compound begins as soon as the tumor has reached an average volume of 100 mm3. Tumor growth was measured two to three times a week and 24 hours after the last treatment by determining the length of two perpendicular axes. Tumor volumes were calculated according to published methods (see Evans et al., Brit. J. Cancer 45, 466-8 [1982]) Antitumor efficacy is determined as the average increase in tumor volume of treated animals divided by the average increase in tumor volume d e untreated animals (controls) and, after multiplication by 100, is expressed as% T / C. The tumor regression (given in%) is presented as the volume

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Smaller medium of the tumor in relation to the average volume of the tumor at the start of treatment. The test compound is administered daily by probe.

As an alternative other cell lines can also be used in the same way, for example:

-the MCF-7 breast adenocarcinoma cell line (ATCC No. HTB 22; see also J. Natl. Cancer Inst. (Bethesda) 51, 1409-16 [1973]);

-the MDA-MB 468 breast adenocarcinoma cell line (ATCC No. HTB 132; see also In Vitro 14, 911-15 [1978]);

-the MDA-MB 231 breast adenocarcinoma cell line (ATCC No. HTB 26; see also J. Natl. Cancer Inst. (Bethesda) 53, 661-74 [1974]);

- the Colo 205 colon carcinoma cell line (ATCC No. CCL 222; see also Cancer Res. 38, 1345-55 [1978]);

-the HCT 116 colon carcinoma cell line (ATCC No. CCL 247; see also Cancer Res. 41, 1751-6 [1981]);

-the DU145 DU 145 prostate carcinoma cell line (ATCC No. HTB 81; see also Cancer Res. 37, 4049-58 [1978]); Y

-the PC-3 PC-3 prostate carcinoma cell line (ATCC No. CRL 1435; see also Cancer Res. 40, 524-34 [1980]).

The inhibition of autophosphorylation of the KDR receptor induced by VEGF can be confirmed with another in vitro experiment in cells: transfected CHO cells, which permanently express the human VEGF receptor (KDR), are seeded in the complete culture medium (with 10 % fetal bovine serum = FCS) in 6-well cell culture plates and incubate at 37 ° C under 5% CO 2, until they show approximately 80% confluence. The compounds that are tested are then diluted in culture medium (without FCS, with 0.1% bovine serum albumin) and added to the cells. (Controls comprise medium without test compounds). After two hours of incubation at 37 ° C, recombinant VEGF is added; the final VEGF concentration is 20 ng / ml). After another five minutes of incubation at 37 ° C, the cells were washed twice with frozen PBS (phosphate buffer) and immediately lysed in 100 µl of lysis regulator solution per well. The lysates were then centrifuged to remove the cell nucleus, and protein concentrations of the supernatants were determined using a commercial protein assay (BIORAD). The lysates can then be used immediately or, if necessary, stored at -20 ° C.

A double phase ELISA is performed to measure phosphorylation of the KDR receptor: a monoclonal antibody for KDR (e.g. Mab 1495.12.14; prepared by H. Towbin) is immobilized on black ELISA plates (OptiPlate ™ HTRF-96 Packard) The plates are then washed and the remaining free protein binding sites are saturated with 1% BSA in PBS. Cell lysates (20 µg of protein per well) are then incubated in these plates overnight at 4 ° C together with an antiphosphotyrosine antibody coupled with alkaline phosphatase (PY20: AP from Transduction Laboratories). The plates were washed again and the binding of the antiphosphotyrosine antibody to the captured phosphorylated receptor, then demonstrated using a luminescent AP substrate (CDP-Star, easy to use, with Emerald II; TROPIX). The luminescence was measured in a Top Count Packard Microplate Scintillation Counter (Top Count). The difference between the positive control signal (stimulated with VEGF) and that of the negative control (not stimulated with VEGF) corresponds to phosphorylation of the KDR receptor induced by VEGF (= 100%). The activity of the tested substances is calculated as% inhibition of KDR receptor phosphorylation induced by VEGF, where the concentration of the substance that induces half of the maximum inhibition is defined as ED50 (effective dose for 50% of inhibition).

A compound according to the invention also inhibits to different degrees other tyrosine kinases involved in the transduction of signals that are measured by trophic factors, for example kinases from the Src family, especially c-Src, Lck, and Fyn kinase; also kinases of the EGF family, for example, c-erbB2 kinase (HER-2), c-erbB3 kinase, c-erbB4 kinase; growth factor kinase receptor such as insulin (IGF-1 kinase), especially members of the PDGF tyrosine kinase receptor family, such as PDGF kinase receptor, GSF-1 kinase receptor, Kit kinase receptor and receptor VEGF kinase; and also serine / threonine kinases, all of which play a role in the regulation of growth and transformation in mammalian cells, including human cells.

Based on these studies, a compound according to the invention shows therapeutic efficacy especially against protein kinase dependent disorders, especially proliferative diseases.

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The usefulness of a compound according to the invention in the treatment of arthritis as an example of an inflammatory rheumatoid or rheumatic disease can be demonstrated as follows:

The well-known rat adjuvant arthritis model (Pearson, Proc. Soc. Exp. Biol. 91, 95-101 (1956)) is used to test the anti-arthritic activity of the compounds according to the invention, or the salts of these. Adjuvant arthritis can be treated using two different dosing schedules: either (i) start time of adjuvant immunization (prophylactic dosing); or from day 15 when the arthritic response is already established (therapeutic dosage). Preferably a therapeutic dosing schedule is used. By comparison, a cyclooxygenase-2 inhibitor, such as 5-promo-2- (4-fluorophenyl) -3- [4- (methylsulfonyl) phenyl] thiophene or diclofenac, is administered in a separate group.

In detail, male Wistar rats (5 animals per group, weighing approximately 200 g, provided by Iffa Credo, France) are injected i.d. (intradermally) at the base of the tail with 0.1 ml of mineral oil containing

0.6 mg of heat-inactivated Mycobacterium tuberculosis, lyophilized. Rats were treated with the test compound (3.10 or 30 mg / kg p.o. once per day), or vehicle (water) from day 15 to day 22 (therapeutic dosing schedule). At the end of the experiment, the inflammation of the tarsal joints was measured by means of a myco-calliper. The percentage of paw inflammation inhibition is calculated based on the arthritic animals treated with vehicle (0% inhibition) and normal animals treated with vehicle (100% inhibition).

Based on these studies, a compound according to the invention is surprisingly suitable for the treatment of inflammatory diseases (especially rheumatic or rheumatoid).

On the basis of their effectiveness as inhibitors of VEGF receptor tyrosine kinase activity, the compounds according to the invention firstly inhibit the growth of blood vessels and therefore, for example, are effective against a number of diseases associated with deregulated angiogenesis, especially diseases caused by ocular neovascularization, especially retinopathies, such as diabetic retinopathy or age-related macular degeneration, psoriasis, hemangioblastoma, such as hemangioma, mesangial cell proliferative disorders, such as acute renal diseases or chronic, for example diabetic nephropathy, malignant nephrosclerosis, thrombotic microangiopathy syndromes or transplant rejection, or especially inflammatory renal disease, such as glomerulonephritis, especially mesangioproliferative glomerulonephritis, hemolytic uremic syndrome, diabetic nephropathy, nephrosclerosis hypertensive, atheroma, arterial restenosis, autoimmune diseases, acute inflammation, fibrotic disorders (for example liver cirrhosis), diabetes, endometriosis, chronic asthma, arterial or post-transplant atherosclerosis, neurodegenerative disorders and especially neoplastic diseases such as leukemia, especially acute lymphoblastic leukemia, acute myeloid leukemia and chronic myeloid leukemia, and other "liquid tumors", especially those that express c-kit, KDR or fit-1, and solid tumors, especially breast cancer, colon cancer, lung cancer (especially lung cancer small cell), prostate cancer or Kaposi's sarcoma. A compound according to the invention inhibits the growth of tumors and is especially suitable for preventing metastatic spread of tumors and the growth of micrometastases.

A compound according to the invention can be administered alone or in combination with one or more therapeutic agents, the possible combination therapy that takes the form of fixed combinations or the administration of a compound of the invention and one or more therapeutic agents that are staggered or provided independently of each other, or the combined administration of fixed combinations and one or more therapeutic agents. A compound according to the invention may additionally or in addition be administered especially for tumor therapy in combination with chemotherapy, radiotherapy, immunotherapy, surgical intervention, or a combination thereof. Long-term therapy is equally possible as is adjuvant therapy in the context of other treatment strategies, as described above. Other possible treatments are therapy to maintain the patient's condition after tumor regression, or even chemopreventive therapy, for example in patients at risk.

The therapeutic agents for the possible combination, are especially one or more antiproliferative, cytostatic or cytotoxic compounds, for example a chemotherapeutic agent or several agents selected from the group that includes, but is not limited to, a polyamine biosynthesis inhibitor, a a protein kinase inhibitor, especially a serine protein / threonine kinase, such as protein kinase C, or a protein tyrosine kinase, such as the EGF tyrosine kinase receptor, for example PKI166, the VEGF tyrosine kinase receptor, for example PTK787, or the PDGF tyrosine kinase receptor, for example STI571, a cytokine, a negative growth regulator, such as TGF-β or IFN-β, an aromatase inhibitor, for example letrozole or anastrozole, an interaction inhibitor of an SH2 domain with a phosphorylated protein, antiestrogens, topoisomerase I inhibitors, such as irinotecan, topoisomerase II inhibitors, agen active agents on microtubules, for example paclitaxel, discodermolide or an epothilone, alkylating agents, antineoplastic antimetabolites, such as gemcitabine or capecitabine, platinum compounds, such as carboplatin or cisplatin, anti-angiogenic compounds, gonadorelin agonists, anti-androgens, bisphosphons , for example AREDIA® or ZOMETA®, and trastuzumab. The structure of active agents, identified by code numbers, genetic or trademarks can be taken from the current edition of the standard compendium "The Merck Index" or from the

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databases, for example international patents (for example IMS World Publications). The corresponding content of these is incorporated in this document by reference.

With the compounds according to the invention mentioned hereinafter, the definitions of the substituents from the general definitions mentioned above, can be reasonably used, for example, to replace more general definitions with more specific definitions or especially with the definitions characterized for being preferred.

On the other hand, the invention relates to the use of a compound according to the invention wherein the radicals and symbols have the meanings as defined above, or a pharmaceutically acceptable salt thereof for the preparation of a pharmaceutical product for treatment. of retinopathy or age-related macular degeneration.

On the other hand, the invention describes a method for the treatment of a neoplastic disease that responds to an inhibition of the tyrosine kinase activity of the VEGF receptor, which comprises the administration of a compound according to the invention or a pharmaceutically acceptable salt. of these, where the radicals and symbols have the meanings, as defined above, in an amount effective against said disease, to a warm-blooded animal that needs such treatment.

On the other hand, the invention describes a method for the treatment of age-related retinopathy or macular degeneration, comprising the administration of a compound according to the invention or a pharmaceutically acceptable salt thereof, wherein the radicals and symbols they have the meanings, as defined above, in an effective amount against said diseases, to a warm-blooded animal that needs such treatment.

The invention relates to a compound of formula I, wherein

Ar is represented by the N-oxide sub-formula Ib,

R1 represents trifluoromethyl,

R2 represents bromine, propyl, propenyl or propynyl.

The invention also relates to the following compounds: N- (4-Chloro-3-trifluoromethyl-phenyl) -2 - [(1-oxy-pyridin-4ylmethyl) -amino] -benzamide, and N- (4-Fluoro- 3-Trifluoromethyl-phenyl) -2 - [(1-oxy-pyridin-4-ylmethyl) -amino] -benzamide, and the tautomers thereof, or a salt of said compound or its tautomer.

A compound of the invention can be prepared by the processes that, although not yet applied for the new compounds of the present invention, are known per se, especially a process characterized in that for the synthesis of a compound of the formula I wherein R2 represents bromine and the remaining symbols R1 and Ar are as defined above for a compound of formula I, a compound of formula II

image 1

wherein R1 and R2 are as defined above, it is reacted with a carbonyl compound of the formula III

image 1

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wherein Ar is as defined above for a compound of formula I, in the presence of a reducing agent,

wherein the initial compounds of formula II and III may also be present with the functional groups in their protected form, if necessary, and / or in the form of salts, provided that a group forming a salt is present and the salt reaction is possible;

wherein any of the protecting groups in a protected derivative of a compound of the formula I is removed;

and, the compound obtained is converted into a compound of formula I, and if desired a free compound of formula I is converted into a salt, or a salt obtained from a compound of formula I, it is converted into the free compound or other salt , and / or a mixture of isomeric compounds of formula I, is separated into the individual isomer.

Alternatively, a compound of formula II can be reacted with a compound of formula III, in the presence of an acid, for example camphor-10-sulfonic acid, in an appropriate solvent similar to toluene or benzene at reflux temperature between approximately 15 minutes and 6 hours providing a bicyclic compound of formula IV,

image 1

wherein R2 represents a hydrogen or halogen and the remaining symbols R1 and Ar are as defined above for a compound of the formula I, bicyclic compound of formula IV, which can also be reacted in an appropriate solvent with triethylsilane and trifluoroacetic acid at a temperature between 60 ° C and 90 ° C for a duration between 4 and 12 hours providing a compound of formula I, wherein R2 represents a hydrogen or halogen and the remaining symbols R1 and Ar are as defined above for a compound of the formula I.

Detailed description of reductive alkylation:

In the more detailed description of the process below, R1, R2 and Ar are as defined for the compounds of formula I, unless otherwise indicated.

The carbonyl compound of formula III may also be present in the form of a reactive derivative; however, free aldehyde or ketone is preferred.

The reactive derivatives of the compounds of formula III are, for example, the corresponding bisulfite adducts or especially semi-acetals, acetals, semicetals or ketals of compounds of formula III with alcohols, for example lower alkanols; or thioacetals or thiocetals of compounds of formula III with mercaptans, for example lower alkane sulfides.

Reductive alkylation is preferably carried out with hydrogenation in the presence of a catalyst, especially a noble metal catalyst, such as platinum or especially palladium, which preferably binds to a carrier material, such as carbon, or a heavy metal catalyst. , such as Raney nickel, at normal pressure

or at pressures between 0.1 to 10 MegaPascals (MPa), or with reduction by means of complex hydrides, such as borohydrides, especially alkali metal cyanoborohydrides, for example sodium cyanoborohydride, in the presence of an appropriate acid, preferably relatively weak acids, such as lower alkanocarboxylic acids, especially acetic acid, or a sulfonic acid, such as p-toluenesulfonic acid; in usual solvents, for example alcohols, such as methanol or ethanol, or ethers, for example cyclic ethers, such as tetrahydrofuran, in the presence or absence of water.

Protective groups

If one or more functional groups, for example carboxy, hydroxy, amino, or mercapto, are or should be protected in a compound of formulas II or III, because they do not take part in the reaction, these are such groups as used

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usually in the synthesis of peptide compounds, and also of cephalosporins and penicillins, also as derivatives of nucleic acid and sugars.

The protecting groups may already be present in the precursors and should protect the functional groups as a function, against unwanted side reactions, such as acylations, etherifications, esterifications, oxidations, sololysis, and similar reactions. It is a characteristic of the protective groups that present themselves easily, i.e. without unwanted side reactions, to eliminate, usually by sololysis, reduction, photolysis or also by enzymatic activity, for example under conditions analogous to physiological conditions, and which are not present in the final products. The specialist knows, or can easily establish, the protective groups that are appropriate with the reactions mentioned above and hereafter.

The protection of such functional groups by said protective groups, the protective groups themselves, and their elimination reactions are described for example in standard reference works, such as JFW McOmie, "Protective Groups in Organic Chemistry", Plenum Press, London and New York 1973, in TW Greene, "Protective Groups in Organic Synthesis", Wiley, New York 1981, in "The Peptides"; Volume 3 (editors: E. Gross and J. Meienhofer), Academic Press, London and New York 1981, in "Methoden der organischen Chemie" (Methods of organic chemistry), Houben Weyl, 4th edition, Volume 15 / l, -Georg Thieme Verlag, Stuttgart 1974, in H.-D. Jakubke and H. Jescheit, "Aminosäuren, Peptide, Proteine" (Amino acids, peptides, proteins), Verlag Chemie, Weinheim, Deerfield Beach, and Basel 1982, and in Jochen Lehmann, "Chemie der Kohlenhydrate; Monosaccharide und Derivate" (Chemistry of carbohydrates: monosaccharides and derivatives), Georg Thieme Verlag, Stuttgart 1974.

Additional stages of the process

The salts of a compound of formula I, with a group forming a salt can be prepared in a manner known per se. The acid addition salts of the compounds of formula I, therefore, can be obtained by treatment with an acid or with an appropriate anion exchange reagent. A salt with two acid molecules (for example a dihalogenide of a compound of formula I), can also be converted into a salt with an acid molecule per compound (for example a monohalogenide); this can be done by heating to a fusion, or for example by heating as a solid under high vacuum at elevated temperature, for example from 130 to 170 ° C, an acid molecule that is expelled by the molecule of a compound of formula I.

The salts can generally be converted to free compounds, for example by treatment with appropriate basic agents, for example with alkali metal carbonates, alkali metal hydrogen carbonates, or alkali metal hydroxides, usually potassium carbonate or sodium hydroxide .

An anthranilic acid amide of formula I, wherein R2 represents a halogen, preferably bromine, can also be reacted according to the following process.

An anthranilic acid amide of formula I, wherein R2 represents a halogen is dissolved in a suitable aromatic solvent such as benzene, toluene or xylene and reacted with a stannan of formula VII,

image 1

wherein R3 is a methyl, in the presence of an appropriate catalyst, preferably tetrakis (triphenylphosphine) palladium (0), at a temperature between 90 ° C and 150 ° C, preferably under an argon atmosphere, between 12 and 36 hours in an appropriate aromatic solvent such as benzene, toluene or xylene.

The obtained of the formula I, wherein R2 represents the alkynyl radical -C≡C-R3, can be transformed into a corresponding alkenyl or alkyl radical by the reactions known in the art.

For example, a compound of formula I, wherein R2 represents the alkynyl radical -C≡C-R3 can be hydrogenated in methanol at atmospheric pressure on Raney nickel at a temperature between 15 ° C and 30 ° C, to provide a compound of formula I, wherein R2 represents a propenyl. Said resulting compound of formula I, wherein R2 represents a propenyl, can also be hydrogenated in methanol at atmospheric pressure over 5% platinum in coal at a temperature between 15 ° C and 30 ° C, to provide a compound of formula I , where R2 represents a propyl.

Abbreviations:

DMF dimethylformamide

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EtOAc ethyl acetate

MS Mass spectrum

RT room temperature

TLC thin layer chromatogram

The following Examples serve to illustrate the invention without limiting the invention in its scope. Temperatures were measured in degrees Celsius (° C). Unless stated otherwise, the reactions are performed at room temperature.

EXAMPLES and Reference Examples

Reference Example 1: 2 - [[6-Methoxy-3-pyridinyl] methyl] amino-N- [4-bromo-3- (trifluoromethyl) phenyl] benzamide (not claimed)

Sodium cyanoborohydride (8.80 g of 95%, 133 mmol) is added in portions for 30 minutes to a stirring mixture of acetic acid (3.8 mL), 6-methoxy-3-pyridinecarboxaldehyde (Fluka, Buchs, Switzerland; 7.80 g , 57 mmol) and 2-amino-N- (4-bromo-3-trifluoromethylphenyl) -benzamide (step 1.2; 13.65 g, 38 mmol) in methanol (380 mL) at 25 ° C. The mixture is stirred for 16 hours. The solvent is evaporated under reduced pressure to provide a residue that is treated with a saturated aqueous solution of sodium bicarbonate (500 mL) and extracted with dichloromethane (3 x 150 mL). The combined extracts are dried (Na2SO4), filtered and the solvent is evaporated under reduced pressure to yield the crude product that is purified by column chromatography on silica gel, eluent 5% EtOAc in dichloromethane and recrystallized from diethyl ether - hexane, to provide the title compound as a beige crystalline solid, mp 101-103 ° 0.

Stage 1.1: 2-Nitro-N- (4-bromo-3-trifluoromethylphenyl) benzamide

A solution of 3-amino-6-bromobenzobifluoride (Fluka, Buchs, Switzerland; 24.0 g, 100 mmol) in EtOAc (240 mL) is added to an aqueous stirred solution of sodium hydroxide (110 mL of 1 M) at room temperature . These stirred solutions are then treated dropwise for 30 minutes with a solution of 2-nitrobenzoyl chloride (Fluka, Buchs, Switzerland; 14.5 mL, 110 mmol) in ETOAc (150 mL). Then, the resulting mixture is stirred for 30 min at room temperature. The mixture is extracted with EtOAc (3 x 100 mL) and the combined extracts are washed sequentially with hydrochloric acid (2 x 100 mL of 2M), water (2 x 100 mL), saturated aqueous sodium bicarbonate solution (2 x 100 mL) and saturated aqueous sodium chloride (1 x 100 mL), dried (MgSO4), filtered and the solvent evaporated under reduced pressure, to produce the crude product, which is purified by recrystallization from EtOAc-hexane , to provide the title compound as a beige crystalline solid, mp 157 158 ° C.

Stage 1 .2: 2-Amino-N- (4-bromo-3-trifluoromethylphenyl) benzamide

A solution of 2-nitro-N- (4-bromo-3-trifluoromethylphenyl) benzamide (intermediate 1a; 32 g, 82 mmol) in methanol (1000 mL) is hydrogenated at atmospheric pressure on Raney nickel (6 g) at 21 ° C. The calculated amount of hydrogen is absorbed after 7 hours. The mixture is filtered and the solvent is evaporated under reduced pressure, to produce the crude product which is purified by recrystallization from diethyl ether-hexane, to provide the title compound as a colorless crystalline solid, m.p. 142-144 ° C.

Reference Example 2: 2- [4-Pyridinylmethyl] amino-N- [4-bromo-3- (trifluoromethyl) phenyl] benzamide (not claimed)

The title compound is prepared, by a method analogous to that described in Example 1, using the intermediate of step 1.2 and 4-pyridinecarboxaldehyde; m.p. 123-124 ° C.

Reference Example 3: 2- [4 Pyridinylmethyl] amino-N- (4-bromophenyl) benzamide (not claimed)

The title compound is prepared, by a method analogous to that described in Example 1, using the intermediate of step 3.2 and 4-pyridinecarboxaldehyde; m.p. 136-137 ° C.

Stage 3.1: 2-Nitro-N- (4-bromophenyl) benzamide

The title compound is prepared analogously to step 1.1, using 4-bromoaniline (Fluka, Buchs, Switzerland); m.p. 202-205 ° C.

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Stage 3.2: 2-Amino-N- (4-bromophenyl) benzamide

The title compound is prepared analogously to step 1.2, using 2-nitro-N- (4-bromophenyl) benzamide (step 3.1); m.p. 139-144 ° C.

Reference Example 4: 2 - [[3 - [(methylamino) carbonyl] -phenyl] methyl] amino-N- [3- (trifluoromethyl) phenyl] -benzamide (not claimed)

The title compound is prepared, by a method analogous to that described in Example 1, using the intermediate of step 4.2 and 3-formyl-N-methyl-benzamide (prepared according to the method described in WO 98/14449) ; m.p. 166-167 ° 0.

Stage 4.1: 2-Nitro-N- [3- (trifluoromethyl) phenyl] benzamide

The title compound is prepared analogously to step 1.1, using 3- (trifluoromethyl) -benzenamine (Aldrich, Buchs, Switzerland); m.p. 134-135 ° C.

Step 4.2: 2-Amino-N - [(3-trifluoromethyl) phenyl) benzamide

The title compound is prepared analogously to step 1.2, using 2-nitro-N - [(3-trifluoromethyl) phenyl) benzamide (step 2.1); m.p. 132-133 ° C.

Reference Example 5: 2 - [[6-Methoxy-3-pyridinyl] methyl] amino-N- [4- (1-propynyl) -3- (trifluoromethyl) phenyl] benzamide (not claimed)

A stirred solution of 2 - [[6-methoxy-3-pyridinyl] methyl] amino-N- [4-bromo-3- (trifluoromethyl) -phenyl] benzamide (Example 1;

3.98 g, 8.3 mmol) in dry toluene (200 mL) is purged with argon for 20 minutes at 25 ° C. Then, tributyl-1-propylstannan (4.1 g of 80%, 9.96 mmol) and tetrakis (triphenylphosphine) palladium (0) (260 mg) are added and the resulting mixture is heated at 100 ° C for 17 hours under an atmosphere Argon The mixture is then cooled, treated with an aqueous solution of sodium hydroxide (85 mL of 0.1 M) and purged with air for 2 hours. The resulting mixture is extracted with EtOAc (3 x 100 mL). The organic phase is washed sequentially with water (2 x 40 mL) and saturated aqueous sodium chloride (1 x 40 mL), dried (Na2SO4), filtered and the solvent evaporated under reduced pressure, to produce the crude product, which is purified by column chromatography on silica gel, eluent 33% EtOAc in hexane and recrystallized from diethyl ether-hexane, to provide the title compound as a pale yellow crystalline solid; m.p. 123-124 ° C.

Reference Example 6: 2- [4-Pyridinylmethyl] amino-N- [4- (1-propyl) -3- (trifluoromethyl) phenyl] -benzamide (not claimed)

The title compound is prepared, by a method analogous to that described in Example 5, using 2- [4-pyridinylmethyl] amino-N- [4-bromo-3- (trifluoromethyl) phenyl] benzamide (Example 2); m, p. 165-166 ° C.

Reference Example 7: 2 [4-Pyridinylmethyl] amino-N- [4- (1-propynyl) phenyl] benzamide (not claimed)

The title compound is prepared, by a method analogous to that described in Example 5, using 2- [4-pyridinylmethyl] amino-N- [4-bromophenyl] benzamide (Example 3); m.p. 147-155 ° C.

Reference Example 8: 2- [4-Pyridinylmethyl] amino-N- [4 - [(Z) -1-propenyl] -3- (trifluoromethyl) -phenyl] benzamide hydrochloride salt (not claimed)

A solution of 2- [4-pyridinylmethyl] amino-N- [4- (1-propynyl) -3- (trifluoromethyl) phenyl] benzamide (Example 6; 0.13 g, 0.32 mmol) in methanol (6 mL) is hydrogenated at atmospheric pressure on Raney nickel (50 mg) at 22 ° C. Hydrogen absorption is achieved after 7 hours. The mixture is then filtered and the solvent is evaporated under reduced pressure, to produce the crude product which is purified by column chromatography on silica gel, eluent 50% EtOAc in dichloromethane to provide the product as an oil. The oil is dissolved in ethanol, acidified with a solution of hydrogen chloride in EtOAC (2M) and diluted with diethyl ether. The resulting precipitate is completely filtered, washed with diethyl ether, dried and purified by recrystallization from diethyl ether-ethanol, to provide the title compound as a beige solid.

Reference Example 9: 2- [4-Pyridinylmethyl] amino-N- [4- (1-propyl) -3- (trifluoromethyl) phenyl] benzamide (not claimed)

A solution of 2- [4-pyridinylmethyl] amino-N- [4 - [(Z) -1-propenyl] -3- (trifluoromethyl) phenyl] benzamide (Example 8; 0.80 g,

1.75 mmol) in methanol (25 mL), hydrogenated at atmospheric pressure over 5% platinum in carbon (0.2 g) at 22 ° C. The calculated amount of hydrogen is absorbed after 12 hours. The mixture is then filtered and the solvent is

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evaporate under reduced pressure, to produce the crude product, which is purified by column chromatography on silica gel, eluent 20% dichloromethane in EtOAc and recrystallized from diethyl ether-hexane, to provide the title compound as a solid. colorless lens; m.p. 134-135 ° C.

Example 10: N- (4-Chloro-3-trifluoromethyl-phenyl) -2 - [(1-oxy-pyridin-4-ylmethyl) -amino] -benzamide

Under an atmosphere of N2, 0.50 g (1.2 mMol) of rac. 3- (4-Chloro-3-trifluoromethyl-phenyl) -2- (1-oxy-pyridin-4-yl) -2,3-dihydro1H-quinazolin-4-one is suspended in 8 ml of 1,2-dichloroethane frozen. Next, 0.47 ml (3.0 mMol) of triethylsilane are added, after 5 min for 0.56 ml (7.2 mMol) of trifluoroacetic acid. The resulting solution is stirred for 7 h at 80 ° C, cooled to RT and diluted with 100 ml of EtOAc. The solution is then washed twice with saturated NaHCO3 solution and brine. The aqueous layers are extracted twice with EtOAc, the organic phases are dried (Na2SO4) and partially concentrated in vacuo until the product crystallizes. Filtration and washing with EtOAc gives the title compound; m.p. 213-214 ° C. The additional product can be obtained from the filtration by column chromatography (SiO2; EtOAc / MeOH 8: 2).

Stage 10.1: rac. 3- (4-Chloro-3-trifluoromethyl-phenyl) -2- (1-oxy-pyridin-4-yl) -2,3-dihydro-1H-quinazolin-4-one

A suspension of 10.04 g (31.9 mMol) of 2-amino-N- [4-chloro-3-trifluoromethyl) phenyl] -benzamide (see preparation WO 00/27820; intermediate 2f) in 80 ml of toluene is prepared in a container with a water separation equipment. 3.93 g (31.9 mMol) of 1-oxy-pyridine-4-carbaldehyde and 23 mg of camphor-10-sulfonic acid are added. The mixture is heated to 120 ° C, for 1 h. After cooling to RT, the reaction mixture is filtered and the residue obtained is washed with toluene and diethyl ether, yielding the title compound; m.p. 261-262 ° C.

Example 11: N- (4-Fluoro-3-trifluoromethyl-phenyl) -2 - [(1-oxy-pyridin-4-ylmethyl) -amino] -benzamide

Under an atmosphere of N2, 1.50 g (3.7 mMol) of rac. 3- (4-Fluoro-3-trifluoromethyl-phenyl) -2- (1-oxy-pyridin-4-yl) -2,3-dihydro1H-quinazolin-4-one, are suspended in 25 ml of frozen dichloromethane. Then 0.83 ml (5.2 mMol) of triethylsilane are added, followed by 1.79 ml (23 mMol) of trifluoroacetic acid. The resulting solution is stirred for 72 h at RT, then another 0.42 ml of triethylsilane are added. After a total of 138 h at RT, the solution is diluted with 300 ml of EtOAc and 300 ml of saturated NaHCO3 solution. The aqueous layer is completely separated and extracted twice with EtOAc. The organic phases were washed with saturated NaHCO3 solution and brine, dried (Na2SO4) and concentrated. The residue is dissolved in methanol, SiO2 is added and the mixture is concentrated. The resulting powder is placed on a chromatographic column (SiO2). Elution with 3: 1 acetone / EtOH gives the title compound; m.p. 182-184 ° C.

Stage 11.1: rac. 3- (4-Fluoro-3-trifluoromethyl-phenyl) -2- (1-oxy-pyridin-4-yl) -2,3-dihydro-1 H-quinazolin-4-one

The condensation of 2.42 g (8.1 mmol) of 2-amino-N- [4-fluoro-3-trifluoromethyl) phenyl] -benzamide (see preparation in WO 00/27820; intermediate 2h) and 980 mg (7.96 mmol) of 1 -oxy-pyridine-4-carbaldehyde in 20 ml of toluene and 5 mg of camphor-10-sulfonic acid analogously to Step 10.1, provides the title compound; m.p. 257-258 ° C.

Example 12: Softgels

5000 soft gelatin capsules, each comprising as an active ingredient 0.05 g of one of the compounds of formula I, mentioned in the preceding Examples, are prepared as follows:

Composition

Active ingredient 250 g

Lauroglycol 2 liters

Preparation process: The powdered active ingredient is suspended in Lauroglykol® (propylene glycol laurate, Gattefossé S.A., Saint Priest, France) and ground in a wet spray to produce a particle size of approximately 1 to 3 µm. 0.419 g portions of the mixture are then introduced into soft gelatin capsules using a capsule filling machine.

REFERENCES CITED IN THE DESCRIPTION

This list of references cited by the applicant is only for the convenience of the reader. It is not part of the European patent document. Even though great care has been taken in collecting references, errors or omissions cannot be excluded and the EPO disclaims all responsibility in this regard.

10 Patent documents cited in the description

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WO 0027820 A [0008] [0080] [0082]

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WO 9814449 A [0071] Non-patent literature cited in the description

• Breier, G. et al. Trends in Cell Biology, 1996, vol. 6, 454-6 [0003] 15 • Shibuya et al. Oncogens, 1990, vol. 5, 519-24 [0026]

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Cancer Research, 1978, vol. 37, 4049-58 [0028]

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Evans et al. Brit. J. Cancer, 1982, vol. 45, 466-8 [0028]

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Bethesda J. Natl. Cancer Inst., 1973, vol. 51, 1409-16 [0029]

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In Vitro, 1978, vol. 14, 911-15 [0029] 20 • J. Natl. Cancer Inst., 1974, vol. 53, 661-74 [0029]

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Cancer Res., 1978, vol. 38, 1345-55 [0029]

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Cancer Res., 1981, vol. 41, 1751-6 [0029]

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Cancer Res., 1978, vol. 37, 4049-58 [0029]

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Cancer Res., 1980, vol. 40, 524-34 [0029] 25 • Pearson. Proc. Soc. Exp. Biol., 1956, vol. 91, 95-101 [0035]

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J. F. W. McOmie. Protective Groups in Organic Chemistry. Plenum Press, 1973 [0055]

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T. W. Greene. Protective Groups in Organic Synthesis. Wiley, 1981 [0055]

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The Peptides Academic Press, 1981, vol. 3 [0055]

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Houben Weyl Methoden der Organischen Chemie. Georg Thieme Verlag, 1974, vol. 15 [0055] 30 • H.-D. Jakubke; H. Jescheit. Aminosäuren, Peptide, Proteine. Verlag Chemie, Weinheim, 1982 [0055]

• Jochen Lehmann. Chemie der Kohlenhydrate; Monosaccharide und Derivate. Georg Thieme Verlag, 1974 [0055]

Claims (5)

1. An anthranilic acid amide of formula I, image 1 where Ar is represented by the N-oxide of sub-formula Ib, image 1 10 Y R1 represents trifluoromethyl, and R2 represents bromine, propyl, propenyl or propynyl,

or a tautomer thereof, or a salt of said anthranilic acid amide or its tautomer.

2. An anthranilic acid amide selected from N- (4-Chloro-3-trifluoromethyl-phenyl) -2 - [(1-oxy-pyridin-4-ylmethyl) -amino] -benzamide, and N- (4-Fluoro-3-trifluoromethyl-phenyl) -2 - [(1-oxy-pyridin-4-ylmethyl) -amino] -benzamide, or a salt of said anthranilic acid amide, or its tautomer.

An anthranilic acid, according to any one of claims 1 to 2, or a tautomer thereof, or a pharmaceutically acceptable salt of said compound, for use in a method for the treatment of the human or animal organism.

Four.

Use of an anthranilic acid amide according to any one of claims 1 to 2, or a tautomer thereof, or a pharmaceutically acceptable salt of said compound, for the preparation of a pharmaceutical product for the treatment of a neoplastic disease.

5.

Use of an anthranilic acid amide according to any one of claims 1 to 2, or a tautomer of

These, or a pharmaceutically acceptable salt of said compound, for the preparation of a pharmaceutical product for the treatment of age-related retinopathy or macular degeneration. 6. A pharmaceutical preparation comprising an anthranilic acid amide according to any one of claims 1 to 2, or a tautomer thereof, or a pharmaceutically acceptable salt of said compound, or a hydrate or solvate thereof, and at least one pharmaceutically acceptable carrier. 10 7. A process for the preparation of an anthranilic acid amide of formula I image 1 wherein R2, R1 and Ar are as defined in claim 1, wherein a compound of formula II image 1 Wherein R1 and R2 are as defined above for formula I, it is reacted with a carbonyl compound of formula III image 1 wherein Ar is represented by sub-formula Ib, as defined in claim 1, in the presence of a reducing agent, 20 wherein, the initial compounds of formula II and III may also be present with the functional groups in their protected form, if necessary, and / or in the form of salts, provided that a salt forming group is present and the salt reaction is possible; wherein any of the protecting groups in a protected derivative of a compound of the formula I is removed; and, the compound obtained is converted into a compound of formula I, and, if so desired, a free compound of formula I, is converted into a salt, or a salt obtained from a compound of formula I, is converted to the compound free or other salt, and / or a mixture of isomeric compounds of formula I, is separated into the individual isomers.