ES2362214T3 - INDAZOLS REPLACED WITH AMIDA AS POLY INHIBITORS (ADP-RIBOSA) POLYMERASE (PARP). - Google Patents

Abstract

The present invention relates to compounds of formula (I): and pharmaceutically acceptable salts, stereoisomers or tautomers thereof which are inhibitors of poly(ADP-ribose)polymerase (PARP) and thus useful for the treatment of cancer, inflammatory discuses, reperfusion injuries, ischemic conditions, stroke, renal failure, cardiovascular diseases, vascular diseases other than cardiovascular diseases, diabetes, neurodegenerative diseases, retroviral infection, retinal damage or skin senescence and UV-induced skin damage, and as chemo and/or radiosensitizers for cancer treatment.

Description

The present invention relates to amide substituted indazoles that are inhibitors of the enzyme poly (ADPribose) polymerase (PARP), formerly known as poly (ADP-ribose) synthase and poly (ADP-ribosyl) transferase. The compounds of the present invention are useful as monotherapies in tumors with specific defects in DNA repair pathways and as enhancers of certain DNA damage agents such as antineoplastic agents and radiotherapy. In addition, the compounds of the present invention are useful for reducing cellular necrosis (in stroke and myocardial infarction), negatively regulating inflammation and tissue lesions, treating retroviral infections and protecting against the toxicity of chemotherapy.

Poly (ADP-ribose) polymerase (PARP) constitutes a super family of eighteen proteins that contain PARP catalytic domains (Bioessays (2004) 26: 1148). These proteins include PARP-1, PARP-2, PARP-3, tankirase-1, tankirase-2, vaultPARP and TiPARP. PARP-1, the founding member, consists of three main domains: an amino (N) -terminal (DBD) DNA binding domain that contains two zinc fingers, the self-modifying domain and a carboxy (C) -terminal catalytic domain .

PARPs are nuclear and cytoplasmic enzymes that cleave NAD + to nicotinamide and ADP-ribose to form branched and long ADP-ribosome polymers in target proteins, including topoisomerases, histones and PARP itself (Biochem. Biophys. Res. Commun. (1998) 245: 1-10).

Poly (ADP-ribosil) ation has been implicated in several biological processes, including DNA repair, gene transcription, cell cycle progression, cell death, chromatin functions and genomic stability.

It has been shown that the catalytic activity of PARP-1 and PARP-2 is rapidly stimulated by DNA chain breaks (see Pharmacological Research (2005) 52: 25-33). In response to DNA damage, PARP-1 binds to single and double DNA notches. Under normal physiological conditions there is minimal PARP activity, however, after DNA damage an immediate activity of PARP activity occurs up to 500 times. Both PARP-1 and PARP-2 detect disruptions of DNA strands acting as notch sensors, which provide rapid signals to stop transcription and recruit the enzymes required for DNA repair at the site of damage. Since radiation therapy and many chemotherapy approaches to cancer therapy work by inducing DNA damage, PARP inhibitors are useful as chemo and radiosensitizers for cancer treatment. It has been indicated that PARP inhibitors are effective in radiosensitization of hypoxic tumor cells (US 5,032,617, US 5,215,738 and US 5,041,653).

Most of the biological effects of PARP are related to this poly (ADP-ribosil) ation process that influences the properties and function of the target proteins; with PAR oligomers which, when cleaved from poly (ADP-ribosyl) proteins, confer different cellular effects; the physical association of PARP with nuclear proteins to form functional complexes and the reduction of the cellular level of its NAD + substrate (Nature Review (2005) 4: 421-440).

In addition to being involved in DNA repair, PARP can also act as a mediator of cell death. Its excessive activation in pathological conditions such as ischemia and reperfusion injury can result in substantial depletion of intercellular NAD +, which can lead to the alteration of several NAD + dependent metabolic pathways and results in cell death (see Pharmacological Research (2005) 52: 44-59). As a result of PARP activation, NAD + levels decrease significantly. Extensive activation of PARP leads to severe depletion of NAD + in cells suffering from massive DNA damage. The short half-life of poly (ADP-ribose) results in a rapid renewal rate, since once poly (ADP-ribose) has formed, it is rapidly degraded by the active poly (ADP-ribose) glycohydrolase (PARG) constitutively. PARP and PARG form a cycle that converts a large amount of NAD + to ADP-ribose, causing a reduction of NAD + and ATP to less than 20% of the normal level. Such a scenario is especially harmful during ischemia when oxygen deprivation has already drastically impaired cellular energy production. It is assumed that subsequent free radical production during reperfusion is a leading cause of tissue damage. Part of the reduction in ATP, which is typical in many organs during ischemia and reperfusion, could be linked to NAD + depletion due to poly (ADP-ribose) renewal. Therefore, PARP inhibition is expected to preserve the level of cellular energy thereby enhancing the survival of ischemic tissues after injury. Compounds that are PARP inhibitors are therefore useful for treating conditions that result from PARP-mediated cell death, including neurological conditions such as stroke, trauma and Parkinson's disease.

PARP inhibitors have been shown to be useful for the specific death of tumors deficient in BRCA1 and BRCA-2 (Nature (2005) 434: 913-916 and 917-921; and Cancer Biology & Therapy (2005) 4: 934-936 ).

PARP inhibitors have been shown to enhance the efficacy of antineoplastic drugs (Pharmacological Research (2005) 52: 25-33), including platinum compounds such as cisplatin and carboplatin (Cancer Chemother Pharmacol (1993) 33: 157-162 and Mol Cancer Ther (2003) 2: 371-382). PARP inhibitors have been shown to increase the anti-tumor activity of topoisomerase I inhibitors such as Irinotecan and Topotecan (Mol Cancer Ther (2003) 2: 371-382; and Clin Cancer Res (2000) 6: 2860-2867) and this is has been demonstrated in in vivo models (J Natl Cancer Inst (2004) 96: 56-67).

PARP inhibitors have been shown to restore susceptibility to temozolomide (TMZ) antiproliferative and cytotoxic effects (see Curr Med Chem (2002) 9: 1285-1301 and Med Chem Rev Online (2004) 1: 144-150). This has been demonstrated in several in vitro models (Br J Cancer (1995) 72: 849-856; Br J Cancer (1996) 74: 1030-1036; Mol Pharmacol (1997) 52: 249-258; Leukemia (1999) 13 : 901-909; Glia (2002) 40: 44-54; and Clin Cancer Res (2000) 6: 2860-2867 and (2004) 10: 881-889) and in vivo models (Blood (2002) 99: 2241- 2244; Clin Cancer Res (2003) 9: 53705379 and J Natl Cancer Inst (2004) 96: 56-67). PARP inhibitors have also been shown to prevent the onset of necrosis induced by selective N3-adenine methylating agents such as MeOSO2 (CH2) -lexitropsin (Me-Lex) (Pharmacological Research (2005) 52: 25-33).

PARP inhibitors have shown that they act as radiation sensitizers. It has been indicated that PARP inhibitors are effective in the radiosensitization of (hypoxic) tumor cells and effective in preventing the recovery of potentially lethal DNA damage tumor cells (Br. J. Cancer (1984) 49 (Suppl. VI ): 34-42; and Int. J. Radiat. Bioi. (1999) 75: 91-100) and sub-lethal (Clin. Oncol. (2004) 16 (1): 29-39) after radiation therapy , presumably because of its ability to prevent the reintegration of DNA chain breakage and affecting several DNA damage signaling pathways.

Also, PARP inhibitors have been shown to be useful for treating acute and chronic myocardial diseases (see Pharmacological Research (2005) 52: 34-43). For example, it has been shown that simple injections of PARP inhibitors have reduced the size of the infarction caused by ischemia and reperfusion of cardiac or skeletal muscle in rabbits. In these studies, an injection of 3-amino-benzamide (10 mg / kg), one minute before occlusion or one minute before reperfusion, caused similar reductions in heart attack size (32-42%) while that 1,5-dihydroxisoquinoline (1 mg / kg), another PARP inhibitor, reduced the size of the infarction to a comparable degree (38-48%). These results make it reasonable to assume that PARP inhibitors could recover a previously ischemic heart or reperfusion injury from skeletal muscle tissue (PNAS (1997) 94: 679-683). Similar findings have been reported in pigs (Eur. J. Pharmacol. (1998) 359: 143-150 and Ann. Thorac. Surg. (2002) 73: 575-581) and in dogs (Shock. (2004) 21: 426 -32).

PARP inhibitors have been shown to be useful for treating certain vascular diseases, septic shock, ischemic injury and neurotoxicity (Biochim. Biophys. Acta (1989) 1014: 1-7; J. Clin. Invest. (1997) 100: 723735) . DNA damage by oxygen radicals that leads to DNA chain breaks, which are subsequently recognized by PARP, is a major contributing factor for such diseases as shown by PARP inhibitor studies (J. Neurosci. Res. (1994 ) 39: 38-46 and PNAS (1996) 93: 4688-4692). It has also been shown that PARP plays a role in the pathogenesis of hemorrhagic shock (PNAS (2000) 97: 1020310208).

PARP inhibitors have been shown to be useful for the treatment of inflammatory diseases (see Pharmacological Research (2005) 52: 72-82 and 83-92).

It has also been shown that effective retroviral infection of mammalian cells is blocked by inhibition of PARP activity. It has been shown that such inhibition of recombinant retroviral vector infections occurs in several different cell types (J. Virology, (1996) 70 (6): 3992-4000). In this way PARP inhibitors have been developed for use in anti-viral therapies and in cancer treatment (WO 91/18591).

In vitro and in vivo experiments have shown that PARP inhibitors can be used for the treatment or prevention of autoimmune diseases such as Type I diabetes and diabetic complications (Pharmacological Research (2005) 52: 60-71).

It has been speculated that PARP inhibition delays the appearance of aging characteristics of human fibroblasts (Biochem. Biophys. Res. Comm. (1994) 201 (2): 665-672 and Pharmacological Research (2005) 52: 93-99) . This may be related to the role PARP plays in controlling the function of telomeres (Nature Gen., (1999) 23 (1): 76-80).

The vast majority of PARP inhibitors to date interact with the nicotinamide binding domain of the enzyme and behave as competitive inhibitors with respect to NAD + (Expert Opin. Ther. Patents (2004) 14: 1531-1551). Structural analogs of nicotinamide, such as benzamide and derivatives were among the first compounds investigated as PARP inhibitors. However, these molecules have a weak inhibitory activity and have other effects unrelated to PARP inhibition. Therefore, there is a need to provide potent inhibitors of the PARP enzyme.

Structurally related PARP inhibitors have been previously described. WO 1999/59973 discloses amide substituted benzene rings fused with 5-membered heteroaromatic rings; WO2001 / 85687 discloses indoles substituted with amide; WO 1997/04771, WO 2000/26192, WO 2000/32579, WO 2000/64878, WO 2000/68206, WO 2001/21615, WO 2002/068407, WO 2003/106430 and WO 2004/096793 disclose substituted benzoimidazoles with amide; WO 2000/29384 discloses benzoimidazoles and indoles substituted by amide; and EP 0879820 discloses benzoxazoles substituted with amide.

It has now surprisingly been discovered that the amide substituted indazoles of the present invention show particularly high levels of inhibition of the activity of poly (ADP-ribose) polymerase (PARP). Therefore the compounds of the present invention are particularly useful as inhibitors of PARP-1 and / or PARP-2. They also show particularly good levels of cellular activity, demonstrating good antiproliferative effects on cell lines deficient in BRCA1 and BRCA2.

The present invention provides compounds of formula I:

image 1

10 in which: R1 is hydrogen or fluorine; and R2 is hydrogen or fluorine;

or pharmaceutically acceptable salts, stereoisomers or tautomers thereof. In one embodiment R1 is hydrogen.

In another embodiment R1 is fluorine. In one embodiment R2 is hydrogen. In another embodiment R2 is fluorine. In one embodiment R1 is hydrogen and R2 is hydrogen or fluorine. In another embodiment R1 is fluorine and R2 is hydrogen or fluorine.

In another embodiment R1 is hydrogen and R2 is hydrogen. In another embodiment R1 is hydrogen and R2 is fluorine. In another embodiment R1 is fluorine and R2 is fluorine. In another embodiment R1 is hydrogen or fluorine and R2 is hydrogen. In another embodiment R1 is hydrogen or fluorine and R2 is fluorine.

The present invention also provides compounds of formula II:

image 1

wherein R1 and R2 are as defined above; 5

10

fifteen

twenty

25

30

or pharmaceutically acceptable salts, stereoisomers or tautomers thereof. The present invention also provides a compound of formula III:

image 1

wherein R1 and R2 are as defined above;

or pharmaceutically acceptable salts or tautomers thereof. The present invention also provides compounds of formula IV:

image 1

wherein R1 and R2 are as defined above;

or pharmaceutically acceptable salts or tautomers thereof.

Preferred identities with reference to formulas II, III and IV are as previously defined for formula I by changing what needs to be changed.

The present invention includes within its scope solvates of the compounds of formula I and salts thereof, for example, hydrates.

The compounds of the present invention may have asymmetric centers, chiral axes and chiral planes (as described in: EL Eliel and SH Wilen, Stereochemistry of carbon Compounds, John Wiley & Sons, New York, 1994, pages 1119-1190), and they appear as racemates, racemic mixtures and as individual diastereomers, with all possible isomers and mixtures thereof, including optical isomers, including all such stereoisomers in the present invention. In addition, the compounds disclosed herein may exist as tautomers and it is intended that both tautomeric forms be encompassed by the scope of the invention, even if only a tautomeric structure is represented.

The compounds may exist in different isomeric forms, all of which are encompassed by the present invention.

The compounds may exist in several different polymorphic forms.

As used herein, C1-6 alkyl represents a branched, single chain and cyclic saturated aliphatic hydrocarbon group containing 1, 2, 3, 4, 5 or 6 carbon atoms. For example, "C1-6alkyl" specifically includes methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, i-butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl and so on. Preferred alkyl groups are methyl and ethyl.

Particular compounds within the scope of the present invention are:

3- {4- [7- (Aminocarbonyl) -2H-indazol-2-yl] phenyl} piperidinium chloride;

2- {4 - [(3R) -Piperidin-3-yl] phenyl} -2H-indazol-7-carboxamide;

2- {4 - [(3S) -Piperidin-3-yl] phenyl} -2H-indazol-7-carboxamide;

3- {4- [7- (Aminocarbonyl) -5-fluoro-2H-indazol-2-yl] phenyl} piperidinium trifluoroacetate; 5

5-Fluoro-2- (3-fluoro-4-piperidin-3-ylphenyl) -2H-indazol-7-carboxamide trifluoroacetate; 3- {4- [7- (Aminocarbonyl) -2H-indazol-2-yl] phenyl} piperidinium trifluoroacetate; 5-Fluoro-2- (4-piperidin-3-ylphenyl) -2H-indazol-7-carboxamide; (3S) -3- {4- [7- (Aminocarbonyl) -2H-indazol-2-yl] phenyl} piperidinium chloride;

5 (3R) -3- {4- [7- (Aminocarbonyl) -2H-indazol-2-yl] phenyl} piperidinium chloride; (R) -5-Fluoro-2- (4-piperidin-3-ylphenyl) -2H-indazol-7-carboxamide; (S) -5-Fluoro-2- (4-piperidin-3-ylphenyl) -2H-indazol-7-carboxamide; (R) -5-Fluoro-2- {3-fluoro-4-piperidin-3-ylphenyl} -2H-indazol-7-carboxamide; (S) -5-Fluoro-2- {3-fluoro-4-piperidin-3-ylphenyl} -2H-indazol-7-carboxamide;

10 and pharmaceutically acceptable salts, bases or tautomers thereof. Stereoisomers thereof of these compounds are also provided. A particular compound of the present invention is: 3- {4- [7- (Aminocarbonyl) -2H-indazol-2-yl] phenyl} piperidinium chloride;

or a pharmaceutically acceptable free base or tautomer thereof. Stereoisomers 15 thereof of this compound are also provided.

A particular compound of the present invention is: 2- {4 - [(3R) -Piperidin-3-yl] phenyl} -2H-indazol-7-carboxamide;

or a pharmaceutically acceptable salt, base or tautomer thereof. Stereoisomers thereof of this compound are also provided.

A particular compound of the present invention is: 2- {4 - [(3S) -Piperidin-3-yl] phenyl} -2H-indazol-7-carboxamide;

or a pharmaceutically acceptable salt, base or tautomer thereof. Stereoisomers thereof of this compound are also provided.

A particular compound of the present invention is: 3- {4- [7- (Aminocarbonyl) -5-fluoro-2H-indazol-2-yl] phenyl} piperidinium trifluoroacetate;

or a pharmaceutically acceptable free base or tautomer thereof. Stereoisomers thereof of this compound are also provided. A particular compound of the present invention is: 5-Fluoro-2- (3-fluoro-4-piperidin-3-ylphenyl) -2H-indazol-7-carboxamide trifluoroacetate; 30 or a pharmaceutically acceptable free base or tautomer thereof. Stereoisomers thereof of this compound are also provided.

A particular compound of the present invention is: (3S) -3- {4- [7- (aminocarbonyl) -2H-indazol-2-yl] phenyl} piperidinium 4-methylbenzenesulfonate;

or a pharmaceutically acceptable free base or tautomer thereof. Stereoisomers thereof of this compound are also provided.

The present free base of compounds of Formula I is included in the present invention, as well as pharmaceutically acceptable salts and stereoisomers thereof. The compounds of the present invention may protonate into the N atom or atoms of an amine and / or N-containing heterocyclic moiety to form a salt. The term "free base" refers to the amine compounds in non-salt form. The pharmaceutically acceptable salts covered

40 not only include the exemplified salts for the specific compounds described herein, but also all the pharmaceutically acceptable salts typical of the free form of compounds of Formula I. The free form of the specific saline compounds described can be isolated using techniques known in The matter. For example, the free form can be regenerated by treating the salt with a suitable dilute aqueous base solution such as dilute aqueous NaOH, potassium carbonate, ammonia and sodium bicarbonate. The free forms may differ from their respective salt forms somewhat in certain physical properties, such as solubility in polar solvents, but the acidic and basic salts are otherwise pharmaceutically equivalent to their respective free forms for the purposes of the invention.

Pharmaceutically acceptable salts of the present compounds can be synthesized from the compounds of the present invention that contain a basic or acidic moiety by conventional chemical procedures. Generally, the salts of the basic compounds are prepared by ion exchange chromatography or by reacting the free base with stoichiometric amounts or with an excess of the organic or inorganic acid that forms the desired salt in a suitable solvent or various combinations of solvents. Similarly, salts of acidic compounds are formed by reactions with the appropriate organic or inorganic base.

Thus, pharmaceutically acceptable salts of the compounds of the present invention include the conventional non-toxic salts of the compounds of the present invention as formed by reaction of a present basic compound with an inorganic, organic or polymeric acid. For example, conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, iodhydric, sulfuric, sulfurous, sulfamic, phosphoric, phosphorous, nitric and the like, as well as salts prepared from organic acids such as acetic, propionic. , succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroximaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanyl, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethanesulfonic, ethanesulfonic, ethanesulfonic, ethanesulfonic , palmitic, gluconic, ascorbic, phenylacetic, aspartic, cinnamic, pyruvic, ethanesulfonic, disulfonic ethane, valeric, trifluoroacetic and the like. Examples of suitable polymer salts include those derived from polymeric acids such as tannic acid, carboxymethyl cellulose. Preferably, a pharmaceutically acceptable salt of the present invention contains 1 equivalent of a compound of formula (I) and 1, 2 or 3 equivalents of an organic or inorganic acid. In one embodiment a pharmaceutically acceptable salt of the present invention contains 2 equivalents of a compound of formula (I) and 1 equivalent of an organic or inorganic acid. More particularly, the pharmaceutically acceptable salts of the present invention are trifluoroacetate, chloride or tosylate salts. More particularly, the pharmaceutically acceptable salts of the present invention are the trifluoroacetate or chloride salts. In one embodiment the salt is trifluoroacetate. In another embodiment the salt is chloride. In another embodiment the salt is tosylate.

The term toluenesulfonic acid can be used interchangeably with 4-methylbenzene sulfonic acid and toluene sulfonates can also be referred to as tosylate salts.

When the compound of the present invention is acidic, "suitable pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic bases that include inorganic bases and organic bases. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, manganous, potassium, sodium, zinc, and the like. Particularly preferred are ammonium, calcium, magnesium, potassium and sodium salts. Salts derived from organic non-toxic pharmaceutically acceptable bases include salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as arginine, lysine, betaine, caffeine, choline, N, N'-dibenzylethylenediamine, ethylamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, diethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, pyropylamine, pipeline, pyrimine, pyrilamine, lysinepropylamine , polyamine, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, dicyclohexylamine, butylamine, benzylamine, phenylbenzylamine, tromethamine and the like resins.

The preparation of the pharmaceutically acceptable salts described above and other typical pharmaceutically acceptable salts are described more specifically in Berg et al (1977) J. Pharm. Sci., ’Pharmaceutical Salts’, 66: 1-19.

It will also be noted that the compounds of the present invention are potentially internal salts of zwitterions, since under physiological conditions a deprotonated acid moiety in the compound, such as a carboxyl group, can be anionic and this electronic charge could then be internally balanced against the cationic charge of a protonated or alkylated basic moiety, such as a quaternary nitrogen atom.

The compounds of the invention can be used in a method of treating the human or animal body by therapy.

The invention provides compounds for use in the treatment or prevention of conditions that can be alleviated by the inhibition of poly (ADP-ribose) polymerase (PARP) (see, for example, Nature Review Drug Discovery (2005) 4: 421-440) .

Therefore, the present invention provides a compound of formula I for use in the manufacture of a medicament for the treatment or prevention of conditions that can be alleviated by the inhibition of poly (ADPribose) polymerase (PARP).

The PARP inhibitors of the present invention are useful for the treatment of the diseases specified in WO 2005/082368.

The compounds of the invention are useful for the treatment of inflammatory diseases, including conditions resulting from organ transplant rejection, such as; chronic inflammatory diseases of the joints, including arthritis, rheumatoid arthritis, osteoarthritis and bone diseases associated with increased bone resorption; inflammatory bowel diseases such as ileitis, ulcerative colitis, Barrett's syndrome and Crohn's disease; inflammatory lung diseases such as asthma, adult respiratory distress syndrome and chronic obstructive airways disease; inflammatory diseases of the eye including corneal dystrophy, trachoma, onchocerciasis, uveitis, sympathetic ophthalmia and endophthalmitis; chronic inflammatory gum diseases, including gingivitis and periodontitis; tuberculosis; leprosy; inflammatory diseases of the kidney including uremic complications, glomerulonephritis and nephrosis; inflammatory skin diseases including sclerodermatitis, psoriasis and eczema; inflammatory diseases of the central nervous system, including chronic demyelinating diseases of the nervous system, multiple sclerosis, AIDS-related neurodegeneration and Alzheimer's disease, infectious meningitis, encephalomyelitis, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis and viral or autoimmune encephalitis; diabetic complications, including, but not limited to, immune complex vasculitis, systemic lupus erythematosus (SLE); inflammatory diseases of the heart such as cardiomyopathy, ischemic heart disease, hypercholesterolemia and atherosclerosis; as well as various other diseases that may have significant inflammatory components, including preeclampsia, chronic liver failure, spinal cord and brain trauma, and multiple organ dysfunction syndrome (MODS) (multiple organ failure (MOF)). The inflammatory disease can also be a systemic inflammation of the body, exemplified by gram positive or gram negative shock, hemorrhagic or anaphylactic shock or shock induced by cancer chemotherapy in response to pro-inflammatory cytokines, for example, shock associated with pro-inflammatory cytokines . Such a shock can be induced, for example, by a chemotherapeutic agent that is administered as a cancer treatment.

Therefore, the present invention provides a compound of formula I for use in the manufacture of a medicament for treating or preventing inflammatory diseases.

The compounds of the present invention may also be useful in the treatment or prevention of reperfusion injuries, resulting from episodes of natural origin and during a surgical procedure, such as intestinal reperfusion injury; myocardial reperfusion injury, reperfusion injury resulting from cardiopulmonary surgical revascularization, aortic aneurysm repair surgery, carotid endarterectomy surgery or hemorrhagic shock; and reoxygenation injury resulting from transplantation of organs such as heart, lung, liver, kidney, pancreas, intestine and cornea.

Therefore, the present invention provides a compound of formula I for use in the manufacture of a medicament for the treatment or prevention of reperfusion injuries.

The compounds of the present invention may also be useful in the treatment or prevention of ischemic conditions, including those resulting from organ transplants, such as stable angina, unstable angina, myocardial ischemia, hepatic ischemia, mesenteric artery ischemia, intestinal ischemia, Critical limb ischemia, chronic critical limb ischemia, cerebral ischemia, acute cardiac ischemia, renal ischemic disease, hepatic ischemic disease, retinal ischemic disorder, septic shock and an ischemic disease of the central nervous system, such as stroke or cerebral ischemia.

Therefore, the present invention provides a compound of formula I for use in the manufacture of a medicament for the treatment or prevention of ischemic conditions.

The present invention provides a compound of formula I for use in the manufacture of a medicament for the treatment or prevention of stroke.

The compounds of the present invention may also be useful for the treatment or prevention of chronic or acute renal failure.

Therefore, the present invention provides a compound of formula I for use in the manufacture of a medicament for the treatment or prevention of renal failure.

The compounds of the present invention may also be useful for the treatment or prevention of vascular diseases other than cardiovascular diseases, such as peripheral arterial occlusion, obliterating thromboangiitis, Reynaud's disease and phenomenon, acrocyanosis, erythromelalgia, venous thrombosis, varicose veins, arteriovenous fistula , lymphedema and lipedema.

Therefore, the present invention provides a compound of formula I for use in the manufacture of a medicament for the treatment or prevention of vascular diseases other than cardiovascular diseases.

The compounds of the present invention may also be useful for the treatment or prevention of cardiovascular diseases such as chronic heart failure, atherosclerosis, congestive heart failure, circulatory shock, cardiomyopathy, heart transplant, myocardial infarction and cardiac arrhythmia, such as atrial fibrillation , supraventricular tachycardia, atrial flutter and paroxysmal atrial tachycardia.

Therefore, the present invention provides a compound of formula I for use in the manufacture of a medicament for the treatment or prevention of cardiovascular diseases.

The compounds of the present invention may also be useful for the treatment and prevention of diabetes mellitus, including Type I diabetes (Insulin-dependent Diabetes Mellitus), Type II diabetes (Non-Insulinodependent Diabetes Mellitus), gestational diabetes, autoimmune diabetes, insulinopathies, diabetes due to pancreatic disease, diabetes associated with other endocrine diseases (such as Cushing's syndrome, acromegaly, pheochromocytoma, glucagonoma, primary aldosteronism or somatostatinoma), Type A insulin resistance syndrome, Type B insulin resistance syndrome, lipatrophic diabetes and diabetes induced by β cell toxins. The compounds of the present invention may also be useful for the treatment or prevention of diabetic complications, such as diabetic cataracts, glaucoma, retinopathy, nephropathy (such as, microalbuminuria and progressive diabetic nephropathy), polyneuropathy, foot gangrene, coronary artery disease. atherosclerotic, peripheral arterial disease, hyperglycemic-hyperosmolar non-ketotic coma, mononeuropathies, autonomic neuropathy, foot ulcers, joint problems and a complication of mucous membrane or skin (such as an infection, a diabetic dermopathy, a candida infection or necrobiosis diabetic lipid, obesity), hyperlipidemia, hypertension, insulin resistance syndrome, coronary artery disease, retinopathy, diabetic neuropathy, polyneuropathy, mononeuropathies, autonomic neuropathy, a foot ulcer, a joint problem, a fungal infection, a bacterial infection na and cardiomyopathy.

Therefore, the present invention provides a compound of formula I for use in the manufacture of a medicament for the treatment or prevention of diabetes.

The compounds of the present invention may also be useful for the treatment or prevention of cancer including solid tumors such as fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing tumor , leiomyosarcoma, rhabdomyosarcoma, colon cancer, colorectal cancer, kidney cancer, pancreatic cancer, bone cancer, breast cancer, ovarian cancer, prostate cancer, esophageal cancer, stomach cancer, oral cancer, nasal cancer, throat, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma choriocarcinoma, seminoma, carc embryonic inoma, Wilms tumor, cervical cancer, uterine cancer, testicular cancer, small cell carcinoma of the lung, bladder carcinoma, lung cancer, epithelial carcinoma, skin cancer, melanoma, neuroblastoma and retinoblastoma; blood-borne cancers such as acute lymphoblastic leukemia ("ALL"), acute lymphocytic B lymphocyte leukemia, acute lymphoblastic T lymphocyte leukemia, acute myeloblastic leukemia ("AML"), acute promyelocytic leukemia ("APL"), monoblastic leukemia acute, acute erythroleukemic leukemia, acute megakaryoblastic leukemia, acute myelomonocytic leukemia, acute non-lymphocytic leukemia, acute undifferentiated leukemia, chronic myelocytic leukemia ("CML"), chronic lymphocytic leukemia ("CLL"), tricoleukemia and multiple myeloma; acute and chronic leukemias such as lymphoblastic, myelogenous, lymphocytic, myelocytic leukemias; lymphomas such as Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease and polycythemia vera; CNS and brain cancers such as glioma, pilocytic astrocytoma, astrocytoma, anaplastic astrocytoma, glioblastoma multiforme, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, cerebular tumor, schwannoma and medulloblastoma.

Therefore, the present invention provides a compound of formula I for use in the manufacture of a medicament for the treatment or prevention of cancer.

The compounds of the present invention can also be used for the treatment of cancer that is deficient in DSB repair activity of Homologous Recombination (RH) dependent DNA (see WO 2006/021801).

The RH-dependent DNA DSB repair path repairs double stranded (DSB) breaks in DNA through homologous mechanisms to reform a continuous DNA helix (Nat. Genet. (2001) 27 (3): 247-254). Components of the RH-dependent DNA DSB repair path include, but are not limited to, ATM (NM000051), RAD51 (NM-002875), RAD51 L1 (NM-002877), RAD51 C (NM-002876), RAD51L3 ( NM-002878), DMC1 (NM-007068), XRCC2 (NM7005431), XRCC3 (NM-005432), RAD52 (NM-002879), RAD54L (NM-003579), RAD54B (NM-012415), BRCA-1 (NM -007295), BRCA-2 (NM-000059), RAD50 (NM-005732), MREI 1A (NM-005590), NBSI (NM-002485), ADPRT (PARP-1), ADPRTL2, (PARP02) CTPS, RPA , RPA1, RPA2, RPA3, XPD, ERCC1, XPF, MMS19, RAD51, RAD51p, RAD51C, RAD51D, DMC1, XRCCR, XRCC3, BRCA1, BRCA2, RAD52, RAD54, RAD50, MRE11, NB51, WRN, BLMKU70, RU80, ATM , ATRCHK1, CHK2, FANCA, FANCB, FANCC, FANCD1, FANCD2, FANCE, FANCF, FANCG, FANCC, FANCD1, FANCD2, FANCE, FANCF, FANCG, RAD1 and RAD9. Other proteins involved in the RH-dependent DNA DSB repair path include regulatory factors such as EMSY (Cell (2003) 115: 523-535).

A cancer that is deficient in repair of RH-dependent DNA DSB may comprise or consist of one or more cancer cells that have a reduced or null capacity to repair DSB of DNA through that route, relative to normal cells ie The activity of the RH-dependent DNA DSB repair path can be reduced or eliminated in one or more cancer cells.

The activity of one or more components of the RH-dependent DNA DSB repair pathway can be eliminated in one or more cancer cells of an individual who has a cancer that is deficient in RH-dependent DNA DSB repair. The components of the RH-dependent DNA DSB repair path are well characterized in the art (see for example, Science (2001) 291: 1284-1289) and include the components listed above.

The present invention provides a compound of formula I for use in the manufacture of a medicament for the treatment or prevention of a cancer that is deficient in repair activity of RH-dependent DNA DSB.

In one embodiment the cancer cells are deficient in the repair activity of RH-dependent DNA DSB of one or more phenotypes selected from ATM (NM-000051), RAD51 (NM-002875), RAD51L 1 (NM002877), RAND51C (NM- 002876), RAD51 L3 (NM-002878), DMC1 (NM-007068), XRCC2 (NM7005431), XRCC3 (NM-005432), RAD52 (NM-002879), RAD54L (NM-003579), RAD54B (NM-012415) , BRCA-1 (NM-007295), BRCA-2 (NM-000059), RAD50 (NM-005732), MREI 1A (NM-005590), NBS1 (NM-002485)), ADPRT (PARP-1), ADPRTL2 , (PARP02) CTPS, RPA, RPA1, RPA2, RPA3, XPD, ERCC1, XPF, MMS19, RAD51, RAD51p, RAD51C, RAD51D, DMC1, XRCCR, XRCC3, BRCA1, BRCA2, RAD52, RAD54, RAD50, MRE11, NB51, WRN, BLMKU70, RU80, ATM, ATRCHK1, CHK2, FANCA, FANCB, FANCC, FANCD1, FANCD2, FANCE, FANCF, FANCG, FANCC, FANCD1, FANCD2, FANCE, FANCF, FANCG, RAD1 and RAD9.

In another embodiment, the cancer cells have a phenotype deficient in BRCA1 and / or BRCA2. Cancer cells with this phenotype may be deficient in BRCA1 and / or BRCA2, that is, the expression and / or activity of BRCA1 and / or BRCA2 can be reduced or eliminated in cancer cells, for example by means of mutation or polymorphism in the acid nucleic coding or by means of amplification, mutation or polymorphism in a gene that encodes a regulatory factor, for example the EMSY gene that encodes a regulatory factor of BRCA2 (Cell (2003) 115: 523-535).

BRCA-1 and BRCA-2 are known tumor suppressors whose wild type alleles are frequently lost in tumors of heterozygous carriers (Oncogene, (2002) 21 (58): 8981-93; Trends Mol Med., (2002) 8 (12): 571-6). The association of BRCA-1 and / or BRCA-2 mutations with breast cancer has been well characterized (Exp Clin Cancer Res., (2002) 21 (3 Supl): 9-12). EMSY gene amplification, which encodes a BRCA-2 binding factor, is also known to be associated with breast cancer and ovarian cancer. Carriers of BRCA-1 and / or BRCA-2 mutations are also at high risk for ovarian, prostate and pancreas cancer. The variation detection in BRCA-1 and BRCA-2 is well known in the art and is described, for example, in EP 699 754, EP 705 903, Genet. Test (1992) 1: 75-83; Cancer Treat Res (2002) 107: 29-59; Neoplasm (2003) 50 (4): 246-50; Ceska Gynekol (2003) 68 (1): 11-16). The determination of amplification of the EMSY BRCA-2 binding factor is described in Cell 115: 523-535. PARP inhibitors have been shown to be useful for the specific death of tumors deficient in BRCA-1 and BRCA-2 (Nature (2005) 434: 913-916 and 917-920).

Therefore, the present invention provides a compound of formula I for use in the manufacture of a medicament for the treatment or prevention of tumors deficient in BRCA-1 or BRCA-2.

In one embodiment, the PARP inhibitors of the present invention can be used in prophylactic therapy for the removal of BRCA-2-deficient cells (see, Cancer Res. (2005) 65: 10145).

The compounds of the present invention may be useful for the treatment or prevention of neurodegenerative diseases, including neurodegeneration related to polyglutamine expansion, Huntington's disease, Kennedy disease, spinocerebelar ataxia, dentotorubral-palidoluisian atrophy (DRPLA), aggregation-related neurodegeneration of proteins, Machado-Joseph disease, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, spongiform encephalopathy, a prion-related disease and multiple sclerosis (MS).

Therefore, the present invention provides a compound of formula I for use in the manufacture of a medicament for the treatment or prevention of neurodegenerative diseases.

The compounds of the present invention may also be useful for the treatment or prevention of retroviral infection (US 5652260), retinal damage (Curr. Eye Res. (2004), 29: 403), skin senescence and UV-induced skin damage ( US5589483 and Biochem. Pharmacol (2002) 63: 921).

The compounds of the invention are useful for the treatment or prevention of premature aging and postpone the onset of age-related cellular dysfunction (Pharmacological Research (2005) 52: 93-99).

The compounds of the present invention can be administered to mammals, preferably humans, alone or in combination with vehicles, excipients, diluents, adjuvants, fillers, buffers, stabilizers, preservatives, pharmaceutically acceptable lubricants, in a pharmaceutical composition, in accordance with practice. conventional pharmaceutical

The compounds of the present invention can be administered to a subject by any convenient route of administration, systemically / peripherally or at the desired site of action, including but not limited to oral (for example by ingestion); topical (including for example transdermal, intranasal, ocular, buccal and sublingual), pulmonary (for example by inhalation or insufflation therapy using, for example an aerosol, for example through the mouth

or the nose); rectal; vaginal; parenteral, (for example, by injection including subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal, subcuticular, intraarticular, subarachnoid and intrasternal) (and implantation of a deposit for example subcutaneously or intramuscularly).

The subject can be a eukaryotic, an animal, a vertebrate animal, a mammal, a rodent (for example a guinea pig, a hamster, a rat, a mouse), murine (for example a mouse), canine (for example a dog) , feline (for example a cat), equine (for example a horse) a primate, ape (for example, a monkey or ape), a monkey (for example, marmoset, baboon), an ape (for example, gorilla, chimpanzee , orangutan, gibbon) or a human being.

The invention also provides pharmaceutical compositions comprising one or more compounds of the present invention and a pharmaceutically acceptable carrier. Pharmaceutical compositions containing the active ingredient may be in a form suitable for oral use, for example, as tablets, troches, tablets, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules or syrups or elixirs. Compositions intended for oral use may be prepared in accordance with any method known in the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preservatives for Provide pharmaceutically elegant and appetizing preparations. The tablets contain the active substance in admixture with pharmaceutically acceptable non-toxic excipients that are suitable for the manufacture of tablets. These excipients may be, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, microcrystalline cellulose, croscarmellose sodium, corn starch or alginic acid; binding agents, for example starch, gelatin, polyvinylpyrrolidone or gum arabic and lubricating agents, for example, magnesium stearate, stearic acid or talc. The tablets may be uncoated or coated by known techniques to mask the unpleasant taste of the drug or delay the disintegration and absorption in the gastrointestinal tract and thereby provide a prolonged action for a longer period. For example, a water-soluble taste masking material such as hydroxypropyl methylcellulose or hydroxypropylcellulose, or a temporary delay material such as ethyl cellulose, cellulose acetate butyrate can be employed.

Formulations for oral use may also be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin or as soft gelatin capsules in which the active principle is mixing with water soluble vehicles such as polyethylene glycol or an oily medium, for example, peanut oil, liquid paraffin or olive oil.

Aqueous suspensions contain the active material in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum arabic, dispersing agents or humectants can be a naturally occurring phosphatide, for example, lecithin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethylene oxyethanol or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. Aqueous suspensions may also contain one or more preservatives, for example, ethyl or n-propyl phydroxybenzoate, one or more coloring agents, one or more flavoring agents and one or more sweetening agents, such as sucrose, saccharin or aspartame.

The oil suspensions may be formulated by suspending the active ingredient in a vegetable oil, for example peanut oil, olive oil, sesame oil or coconut oil or in mineral oil such as liquid paraffin. The oil suspensions may contain a thickening agent, for example beeswax, hard paraffin

or cetyl alcohol. Sweetening agents such as those set forth above and flavoring agents may be added to provide an appetizing oral preparation. These compositions can be preserved by the addition of an antioxidant such as butylated hydroxyanisole or alpha-tocopherol.

Dispersible powders and granules suitable for the preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a wetting or dispersing agent, suspending agent and one or more preservatives. Suitable wetting or dispersing agents and suspending agents are exemplified by those already mentioned above. Additional excipients may also be present, for example sweetening, flavoring and coloring agents. These compositions can be preserved by the addition of an antioxidant such as ascorbic acid.

The pharmaceutical compositions of the invention may also be in the form of oil-in-water emulsions. The oil phase may be a vegetable oil, for example olive oil or peanut oil or a mineral oil, for example liquid paraffin or mixtures thereof. Suitable emulsifying agents may be naturally occurring phosphatides, for example soy lecithin and partial esters or esters derived from fatty acids and hexitol anhydrides, for example sorbitan monooleate and condensation products of said partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate. Emulsions may also contain sweetening agents, flavorings, preservatives and antioxidants.

Syrups and elixirs can be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain an emollient, a preservative, flavoring and coloring agents and antioxidants.

The pharmaceutical compositions may be in the form of sterile injectable aqueous solutions. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution.

The sterile injectable preparation may also be a sterile injectable water-in-water microemulsion in which the active ingredient dissolves in the oil phase. For example, the active substance can be dissolved first in a mixture of soybean oil and lecithin. The oily solution is then introduced into a mixture of water and glycerol and processed to form a microemulsion.

Injectable solutions or microemulsions can be introduced into the bloodstream of a patient by injection of local embolate. Alternatively, it may be advantageous to administer the solution or microemulsion in such a manner that a constant concentration of the present compound is maintained. To maintain such a constant concentration, a continuous intravenous delivery device can be used. An example of such a device is the Deltec CADD-PLUS ™ Model 5400 IV pump.

The pharmaceutical compositions may be in the form of a sterile injectable oily or aqueous suspension for intramuscular and subcutaneous administration. This suppression can be formulated according to the known technique using suitable wetting or dispersing agents and suspending agents mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. In addition, sterile fixed oils are conventionally employed as a solvent or suspending medium. Any insipid fixed oil including synthetic mono or diglycerides can be used for this purpose. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

The compounds of Formula I can also be administered in the form of suppositories for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at ordinary temperatures but liquid at the rectal temperature and will therefore be fused in the rectum to release the drug. Such materials include cocoa butter, glycerinated gelatin, hydrogenated vegetable oils, mixtures of polyethylene glycols of various molecular weights and fatty acid esters of polyethylene glycol.

For topical use, creams, ointments, petrolatum, solutions or suspensions, etc., containing the compound of Formula I are used (For the purposes of the present application, topical application will include mouth washes and gargles).

The compounds for the present invention can be administered intranasally by topical use of suitable intranasal vehicles and delivery devices or by transdermal routes, using the forms of transdermal skin patches well known to those skilled in the art. To administer in the form of a transdermal delivery system, the administration of dosage will, of course, be continuous rather than intermittent throughout the dosing regime. The compounds of the present invention can also be supplied as a suppository using bases such as cocoa butter, glycerinated gelatin, hydrogenated vegetable oils, mixtures of polyethylene glycols of various molecular weights and fatty acid esters of polyethylene glycol.

When a compound according to the present invention is administered to a subject, the dosage level selected will depend on a variety of factors including, but not limited to, activity of the particular compound, the severity of the individual symptoms, the route of administration, the time of administration, the rate of excretion of the compound, the duration of treatment, other drugs, compounds and / or materials used in combination and the age, sex, weight, condition, general health and previous medical history of the patient. The amount of compound and route of administration will ultimately be at the discretion of the physician; although the dosage will generally be to achieve local concentrations at the site of action that achieves the desired effect without causing substantial harmful or deleterious side effects.

Administration can be performed in vivo in one dose, continuously or intermittently (for example in divided doses at appropriate intervals) throughout the course of treatment. Methods for determining the most effective means and dosage of administration are well known to those skilled in the art and will vary with the formulation used for therapy, the purpose of therapy, the target cell being treated and the subject being treated. Single or multiple administrations can be carried out by selecting the dose level and standard by the attending physician.

In general, an adequate dose of the active compound is in the range of about 100 µg to about 250 mg per kilogram of the subject's body weight per day. When the active compound is a salt, an ester, prodrug or the like, the compound administered is calculated based on the parent compound and thus the actual weight to be used is proportionally increased.

The present compounds are also useful in combination with antineoplastic agents or chemotherapeutic agents.

The compounds of the present invention may be useful as chemo and radiosensitizers for cancer treatment. They are useful for the treatment of mammals that have previously undergone or are currently undergoing cancer treatment. Such prior treatments include chemotherapy, radiation therapy, surgery or previous immunotherapy, such as cancer vaccines.

Therefore, the present invention provides a combination of a compound of formula I and an antineoplastic agent for simultaneous, separate or sequential administration.

The present invention also provides a combination of a compound of formula I, radiotherapy and other chemotherapeutic agent for simultaneous, separate or sequential administration.

The present invention also provides a compound of formula I for use in the manufacture of a medicament for use as an accessory in cancer therapy or to enhance tumor cells by combination with ionizing radiation or chemotherapeutic agents.

The present invention also provides the use of a compound of formula I in the manufacture of a medicament for use as an accessory in cancer therapy or to enhance tumor cells by combination with ionizing radiation and other chemotherapeutic agents. The compounds can also be used in combination with ionizing radiation and other chemotherapeutic agents.

In combination therapy the compounds of the present invention can be administered before (for example, 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours , 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks or 12 weeks before), simultaneously with or after (for example, 5 minutes, 15 minutes, 30 minutes , 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks or 12 weeks later) the administration of the other antineoplastic agent to a subject in need. In various embodiments, the present compounds and another antineoplastic agent are administered 1 minute apart, 10 minutes apart, 30 minutes apart, less than 1 hour apart, 1 hour to 2 hours apart, 2 hours to 3 hours. difference, 3 hours to 4 hours apart, 4 hours to 5 hours apart, 5 hours to 6 hours apart, 6 hours to 7 hours apart, 7 hours to 8 hours apart, 8 hours to 9 hours apart, 9 hours to 10 hours apart, 10 hours to 11 hours apart, 11 hours to 12 hours apart, no more than 24 hours apart or no more than 48 hours apart.

The compounds of the present invention and the other antineoplastic agent can act additively or synergistically. A synergistic combination of the present compounds and another antineoplastic agent could allow the use of lesser dosages of one or both of these agents and / or less frequent dosages of one or both of the present compounds and other antineoplastic agents and / or administer the less agents frequently it can reduce any toxicity associated with the administration of the agents to a subject without reducing the effectiveness of the agents in the treatment of cancer. In addition, a synergistic effect could result in an improvement in the efficacy of these agents in the treatment of cancer and / or the reduction of any adverse or unwanted side effects associated with the use of any agent alone.

Examples of cancer agents or chemotherapeutic agents for use in combination with the compounds of the present invention can be found in Cancer Principles and Practice of oncology by V.T. Devita and S. Hellman (editors), 6th edition (February 15, 2001), Lippincott Williams & Wilkins Publishers. One skilled in the art would be able to differentiate which combinations of agents would be useful based on the particular characteristics of the drugs and the cancer involved. Such antineoplastic agents include, but are not limited to, the following: HDAC inhibitors, estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic / cytostatic agents, antiproliferative agents, prenylprotein transferase inhibitors, HMG- inhibitors CoA reductase, HIV protease inhibitors, reverse transcriptase inhibitors and other angiogenesis inhibitors, cell proliferation and survival signaling inhibitors, apoptosis inducing agents and agents that interfere with cell cycle control points. The present compounds are particularly useful when co-administered with radiotherapy.

Examples of "HDAC inhibitors" include hydroxamic suberylanilide acid (SAHA), LAQ824, LBH589,

PXD101, MS275, FK228, valproic acid, butyric acid and CI-994.

"Estrogen receptor modulators" refers to compounds that interfere with or inhibit estrogen binding with the receptor, regardless of mechanism. Examples of estrogen receptor modulators include, but are not limited to, tamoxifen, raloxifene, idoxifene, LY353381, LY117081, toremifene, fulvestrant, 4- [7- (2,2-dimethyl-1-oxopropoxy-4-methyl-2 - [4- [2- (1-Piperidinyl) ethoxy] phenyl] -2H-1-benzopyran-3-yl] -phenyl-2,2-dimethylpropanoate, 4,4'-dihydroxybenzophenone-2,4-dinitrophenyl-hydrazone and SH646 .

"Androgen receptor modulators" refers to compounds that interfere with or inhibit androgen binding with the receptor, regardless of mechanism. Examples of androgen receptor modulators include finasteride and other 5α-reductase, nilutamide, flutamide, bicalutamide, liarozole and abiraterone acetate inhibitors.

"Retinoid receptor modulators" refers to compounds that prevent or inhibit retinoid binding with the receptor, regardless of mechanism. Examples of such retinoid receptor modulators include bexarotene, tretinoin, 13-cis-retinoic acid, 9-cis-retinoic acid, α-difluoromethylornitine, ILX23-7553, trans-N- (4'hydroxyphenyl) retinamide and N-4- carboxyphenyl retinamide.

"Cytotoxic / cytostatic agents" refers to compounds that cause cell death or inhibit cell proliferation primarily by directly preventing cell functioning or inhibiting or interfering with cell mitosis, including alkylating agents, tumor necrosis factors, interleavers, activatable compounds by hypoxia, microtubule stabilizing agents / microtubule inhibitors, mitotic kinesin inhibitors, kinase inhibitors involved in mitotic progression, antimetabolites, biological response modifiers; hormonal / anti-hormonal therapeutic agents, hematopoietic growth agents, monoclonal antibody target therapeutic agents, topoisomerase inhibitors, proteasome inhibitors and ubiquitin ligase inhibitors.

Examples of cytotoxic agents include, but are not limited to, cyclophosphamide, chlorambucil carmustine (BCNU), lomustine (CCNU), busulfan, treosulfan, sertenef, caquectin, ifosfamide, tasonermin, lonidamine, carboplatin, altretamine, prednimustine, dibromustine, dibromustine, dibromustine, dibromustine, dibromustine, dibromustine, dibromustine, dibromustine, dibromustine, dibromothinitol, dibromothinitol, dibromothinitol Nedaplatin, Aroplatin, Oxaliplatin, Temozolomide, Methyl Methanesulfonate, Procarbazine, Dacarbazine, Heptaplatin, Estramustine, Improsulfan Tosylate, Trophosphamide, Nimustine, Dibrospidium Chloride, Pumitepa, Lobaplatin, Satraplatin-Cyrimidin, Cyrimidin-Cyrimine methyl-pyridin) platinum, benzylguanine, glufosfamide, GPX100, (trans, trans, trans) -bis-mu- (hexane-1,6-diamine) -mu- [diamin-platinum (II)] bis [diamine (chlorine) platinum (II)] tetrachloride, diarizidinyl spermine, arsenic trioxide, 1- (11-dodecylamino-10-hydroxyundecyl) -3,7-dimethylxanthine, zorubicin, idarubicin, daunorubicin, bisantrene, mitoxantrone, pyrarubicin, pinavaline bicin, amrubicin, doxorubicin, epirubicin, pyrarubicin, antineoplaston, 3'-desamino-3'-morpholino-13-deoxo-10-hydroxycarminomycin, annamicin, galarubicin, elinafide, MEN 10755 and 4-demethoxy-3-azaminoxy-3 -4-methylsulfonyl-daunorubicin (see WO 00/50032). Additional examples include Raf kinase inhibitors (such as Bay43-9006) and mTOR inhibitors (such as Wyeth CCI-779 and Ariad AP23573). Additional examples are PI3K inhibitors (for example LY294002).

In one embodiment the compounds of the present invention can be used in combination with alkylating agents.

Examples of alkylating agents include, but are not limited to, nitrogen mustards: cyclophosphamide, ifosfamide, triphosphamide and chlorambucil; nitrosoureas: carmustine (BCNU) and lomustine (CCNU); alkylsulfonates: busulfan and threosulfan; triazenos: dacarbazine, procarbazine and temozolomide; complexes containing platinum: cisplatin, carboplatin, aroplatin and oxaliplatin.

In one embodiment, the alkylating agent is dacarbazine. Dacarbazine can be administered to a subject in dosages ranging from approximately 150 mg / m2 (of the body surface area of a subject) to approximately 250 mg / m2. In another embodiment, dacarbazine is administered intravenously to a subject once a day for five consecutive days at a dose ranging from about 150 mg / m2 to about 250 mg / m2.

In one embodiment, the alkylating agent is procarbazine. Procarbazine can be administered to a subject at dosages ranging from about 50 mg / m2 (of a subject's body surface area) to about 100 mg / m2. In another embodiment, procarbazine is administered intravenously to a subject once a day for five consecutive days at a dose ranging from about 50 mg / m2 to about 100 mg / m2.

In one embodiment, the alkylating agent is temozolomide. Temozolomide can be administered to a subject at dosages ranging from approximately 150 mg / m2 (of the body surface area of a subject) to approximately 200 mg / m2. In another embodiment, temozolomide is administered orally to an animal once a day for five consecutive days at a dose ranging from about 150 mg / m2 to about 200 mg / m2.

Examples of anti-mitotic agents: alocolchicine, halicondrine B, colchicine, colchicine derivative, dolstatin 10, maitansin, rhizoxin, thiocolchicine and trityl cysteine.

An example of an activatable hypoxia activatable compound is tirapazamine.

Examples of proteasome inhibitors include but are not limited to lactacistine, bortezomib, epoxomycin and peptide aldehydes such as MG 132, MG 115 and PSI.

Examples of microtubule inhibitors / microtubule stabilizers include paclitaxel, vindesine sulfate, vincristine, vinblastine, vinorelbine, 3 ', 4'-didehydro-4'-deoxy-8'-norvincaleucoblastine, docetaxol, rhizoxin, dolastatin, isethionate mivobulin, auristatin, cemadotine, RPR109881, BMS184476, vinflunine, cryptophytic, 2,3,4,5,6-pentafluoro-N- (3-fluoro-4-methoxyphenyl) benzene sulfonamide, anhydrovinblastine, N, N-dimethyl-L- Valyl-L-Valyl-N-methyl-L-Valyl-L-prolyl-L-proline-t-butylamide, TDX258, epothilones (see, for example, US Pat. Nos. 6,284,781 and 6,288,237) and BMS188797.

Some examples of topoisomerase inhibitors are topotecan, hicaptamine, irinotecan, rubitecan, exatecan, gimetecan, diflomotecan, silyl-camptothecins, 9-aminocamptothecin, camptothecin, chrynatol, mitomycin C, 6-ethoxypropionyl-3 ', 4'-O-exidene cartreusin, 9-methoxy-N, N-dimethyl-5-nitropyrazolo [3,4,5-kl] acridine-2 (6H) propanamine, 1-amino-9-ethyl-5-fluoro-2,3-dihydro- 9-hydroxy-4-methyl-1H, 12H-benzo [de] pyrano [3'4 ': b, 7] indolizino [1,2b] quinolin-10,13 (9H, 15H) dione, lurtotecan, 7- [ 2- (N-isopropylamino) ethyl] - (20S) camptothecin, BNP1350, BNP11100, BN80915, BN80942, etoposide phosphate, teniposide, sobuzoxane, 2'-dimethylamino-2'-deoxy-etoposide, GL331, N- [2- (dimethylamino) ethyl] -9-hydroxy-5,6-dimethyl-6H-pyrido [4,3-b] carbazol-1-carboxamide, asulacrine, (5a, 5aB, 8aa, 9b) -9- [2- [ N- [2- (dimethylamino) ethyl] -N-methylaminium] ethyl] -5- [4-hydroxy-3,5-dimethoxyphenyl] -5,5a, 6,8,8a, 9-hexohydrofuro (3 ', 4': 6.7) naphtho (2,3-d) -1,3-dioxol-6-one, 2,3- (methylenedioxy) -5-methyl-7-hydroxy-8-methoxybenzo [c] phenanthridinium, 6,9-bis [(2-aminoethyl) amino] benzo [g] isoguinolin-5,10-dione, 5- (3-aminopropylamino) -7,10-dihydroxy-2- (2-hydroxyethylaminomethyl) - 6H-pyrazolo [4,5,1-de] acridin-6-one, N- [1- [2 (diethylamino) ethylamino] -7-methoxy-9-oxo-9H-thioxanten4-ylmethyl] formamide, N- ( 2- (dimethylamino) ethyl) acridin-4-carboxamide, 6 - [[2- (dimethylamino) ethyl] amino] -3-hydroxy-7H- indene [2,1-c] quinolin-7-one and dimesna; non-camptothecin topoisomerase I inhibitors such as indolocarbazoles; and dual topoisomerase I and II inhibitors such as benzophenazines, XR20 115761MLN 576 and benzopyridoindoles.

In one embodiment, the topoisomerase inhibitor is irinotecan. Irinotecan can be administered to a subject in dosages ranging from approximately 50 mg / m2 (of the body surface area of a subject) to approximately 150 mg / m2. In another embodiment, irinotecan is administered intravenously to a subject once a day for five consecutive days at a dose ranging from about 50 mg / m2 to about 150 mg / m2 on days 1-5, then again via intravenously once a day for five consecutive days on days 28-32 at a dose that ranges from about 50 mg / m2 to about 150 mg / m2, then again intravenously once a day for five consecutive days on days 55- 59 at a dose that ranges from about 50 mg / m2 to about 150 mg / m2.

Examples of mitotic kinesin inhibitors and in particular KSP human mitotic kinesin are described in PCT Publications WO 01/30768, WO 01/98278, WO 02/056880, WO 03 / 050.064, WO 03 / 050.122, WO 03 /049.527, WO 03 / 049.679, WO 03 / 049.678, WO 03/039460, WO 03/079973, WO 03/099211, WO 2004/039774, WO 03/105855, WO 03/106417, WO 2004/087050, WO 2004 / 058700, WO 2004/058148 and WO 2004/037171 and United States applications US 2004/132830 and US 2004/132719. In one embodiment, mitotic kinesin inhibitors include, but are not limited to KSP inhibitors, MLKP1 inhibitors, CENP-E inhibitors, MCAK inhibitors, Kifl4 inhibitors, Mphosph1 inhibitors and Rab6-KIFL inhibitors.

"Kinase inhibitors involved in mitotic progression" include, but are not limited to, aurora kinase inhibitors, Polo type kinase inhibitors (PLK) (in particular PLK-1 inhibitors), bub-1 inhibitors and bub-R1 inhibitors .

"Antiproliferative agents" include DNA oligonucleotides and antisense RNAs such as G3139, ODN698, RVASKRAS, GEM231 and INX3001 and antimetabolites such as enocitabine, carmofur, tegafur, pentostatin, doxifluridine, trimetrexate, cyclorabine phosphoacetate, oxytodabine, cathodochinoacetate, oxytodabine, cathode citrate, oxytodabine, cathode citrate, oxytorabine, cathode citrate, oxytobine phosteabine, raltitrexed, paltitrexid, emitefur, thiazofurin, decitabine, nolatrexed, pemetrexed, nelzarabine, 2'-deoxy-2'methylidenecitidine, 2'-fluoromethylene-2'-deoxycytidine, N- [5- (2,3-diofur) ) sulfonyl] -N '- (3,4-dichlorophenyl) urea, N6 [4-deoxy-4- [N2- [2 (E), 4 (E) -tetradecadienoyl] glycylamino] -L-glycerol-BL-hand -heptopiranosyl] adenine, aplidine, ecteinascidin, troxacitabine, 4- [2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimidino [5,4-b] [1,4] thiazine- 6-yl- (S) -ethyl] 2,5-thienoyl-L-glutamic, aminopterin, 5-flurouracil, alanosine, 11-acetyl-8- (carbamoyloxymethyl) -4-formyl6-methoxy-14-oxa acid ester -1,11-diazatetracycle (7.4.1.0.0) -tetradeca-2,4,6- trien-9-yl acetic acid, swainsonin, lometrexol, dexrazoxane, methioninase, 2’-cyano-2’-deoxy-N4-palmitoyl-1-B-D-arabinofuranosyl cytosine and 3-aminopyridine-2-carboxaldehyde thiosemicarbazone.

Examples of monoclonal antibody target therapeutic agents include therapeutic agents that have cytotoxic agents or radioisotopes bound to a target cell specific or cancer cell specific monoclonal antibody. Examples include, Bexxar.

"HMG-CoA reductase inhibitors" refers to 3-hydroxy-3-methylglutaryl-CoA reductase inhibitors. Examples of HMG-CoA reductase inhibitors that may be used include but are not limited to lovastatin (MEVACOR®; see U.S. Patent No. 4,231,938, 4,294,926, and 4,319,039), simvastatin (ZOCOR®; see U.S. Patent No. 4,444,784, 4,820,850 and 4,916,239), pravastatin (PRAVACHOL®; see U.S. Patent No. 4,346,227, 4,537859, 4,410,629, 5,030,447 and 5,180,589), Fluvastatin (LESCOL®; see U.S. Patent No. 5,354,772, 4,911,165, 4,929,437, 5,189,164, 5,118,853, 5,290,946 and 5,356,896) and atorvastatin (LIPITOR ®; see U.S. Patent No. 5,273,995, 4,681 .893, 5,489,691 and 5,342,952). The structural formulas of these and other additional HMG-CoA reductase inhibitors that can be used in the present methods are described on page 87 of M. Yalpani, "Lowering Drugs Cholesterol", Chemistry and Industry, p. 85-89 (February 5, 1996) and United States Patents No. 4,782,084 and

4,885,314. The term "HMG-CoA reductase inhibitor" as used herein includes all pharmaceutically acceptable forms of lactone and open acid (i.e., when the lactone ring is opened to form the free acid) as well as salt and ester forms. of compounds having HMG-CoA reductase inhibitory activity and therefore the use of such forms of salts, esters, open acid and lactone is included within the scope of the present invention.

"Pentyl protein transferase inhibitor" refers to a compound that inhibits any one or any combination of prenil protein transferase enzymes, including farnesyl protein transferase (FPTase), geranylgeranyl protein transferase type I (GGPTase-I) and geranylgeranyl -protein transferase type II (GGPTase-II, also called Rab GGPTase).

Examples of prenyl protein transferase inhibitors can be found in the following publications and patents: WO 96/30343, WO 97/18813, WO 97/21701, WO 97/23478, WO 97/38665, WO 98/28980, WO 98 / 29119, WO 95/32987, United States Patent No. 5,420,245, United States Patent No. 5,523,430, United States Patent No. 5,532,359, United States Patent No. 5,510,510, United States Patent No. 5,589. 485, U.S. Patent No. 5,602,098, European Patent Publication 0 618 221, European Patent Publication 0 675 112, European Patent Publication 0 604 181, European Patent Publication 0 696 593, WO 94/19357, WO 95/08542, WO 95/11917, WO 95/12612, WO 95/12572. WO 95/10514, U.S. Patent No. 5,661,152, WO 95/10515, WO 95/10516, WO 95/24612, WO 95/34535, WO 95/25086, WO 96/05529, WO 96/06138, WO 96/06193, WO 96/16443, WO 96/21701, WO 96/21456, WO 96/22278, WO 96/24611, WO96 / 24612, WO 96/05168, WO 96/05169, WO 96/00736, Patent No. 5,571,792, WO 96/17861, WO 96/33159, WO 96/34850, WO 96/34851, WO 96/30017, WO 96/30018, WO 96/30362, WO 96/30363, WO 96/1111, WO 96/31477, WO 96/31478, WO 96/31501, WO 97/00252, WO 97/03047, WO 97/03050, WO 97/04785, WO 97/02920, WO 97/17070, WO 97/23478, WO 97/26246, WO 97/30053, WO 97/44350, WO 98/02436 and U.S. Patent No. 5,532,359. For an example of the role of a prenyl protein transferase inhibitor in angiogenesis see European J. of Cancer (1999), 35 (9): 1394-1401.

"Angiogenesis inhibitors" refers to compounds that inhibit the formation of new blood vessels, regardless of mechanism. Examples of angiogenesis inhibitors include, but are not limited to, tyrosine kinase inhibitors, such as inhibitors of the tyrosine kinase receptor Flt-1 (VEGFR1) and Flk-1 / KDR (VEGFR2), epidermis-derived growth factor inhibitors, fibroblast derivatives or platelet derivatives, MMP inhibitors (matrix metalloprotease), integrin blockers, interferon α, interleukin-12, pentosan polysulfate, cyclooxygenase inhibitors, including nonsteroidal anti-inflammatory drugs (NSAIDs) such as aspirin and ibuprofen as well as selective inhibitors of cyclooxygenase 2 such as celecoxib and rofecoxib (PNAS (1992) 89: 7384; JNCI (1982) 69: 475; Arch. Opthalmol. (1990) 108: 573; Anat. Rec. (1994) 238: 68; FEBS Letters ( 1995) 372: 83; Clin, Orthop. (1995) 313: 76; J. Mol. Endocrinol. (1996) 16: 107; Jpn. J. Pharmacol. (1997) 75: 105; Cancer Res. (1997) 57 : 1625 (1997); Cell (1998) 93: 705; Intl. J. Mol. Med. (1998) 2: 715; J. Biol. Chem. (1999) 274: 9116)), anti-inflation Steroidal amatoriums (such as corticosteroids, mineralocorticoids, dexamethasone, prednisone, prednisolone, methylpred, betamethasone), carboxyamidotriazole, combretastatin A-4, squlamine, 6-Ocloroacetyl-carbonyl) -fumagilol, thalidomide, anginatatin-angiotensin, angiostatin-1, anginatatin (see J. Lab. Clin. 105: 141 Med (1985) -145) and antibodies to VEGF (see Nature Biotechnology (1999) 17: 963-968; Kim et al (1993) -844; WO 00/44777; and WO 00/61186).

Other therapeutic agents that modulate or inhibit angiogenesis and can also be used in combination with the compounds of the present invention include agents that modulate or inhibit coagulation and fibrinolysis systems (see a review in Clin. Chem. La Med. (2000) 38 : 679-692). Examples of such agents that modulate or inhibit coagulation and fibrinolysis pathways include, but are not limited to, heparin (see Thromb. Haemost. (1998) 80: 10-23), low molecular weight heparins, and carboxypeptidase U inhibitors (also known as inhibitors of the prothrombin activatable fibrinolysis inhibitor [TAFIa]) (see Thrombosis Res. (2001) 101: 329-354). TAFIa inhibitors have been described in PCT Publication WO 03/013,526 and U.S. Serial No. 60 / 349,925 (filed January 18, 2002).

"Agents that interfere with cell cycle control points" refers to compounds that inhibit protein kinases that transduce signals from cell cycle control points, thereby sensitizing the cancer cell to DNA damage agents. Such agents include ATR inhibitors, ATM chk1 and Chk2 kinases and cdk and cdc kinase inhibitors and are specifically exemplified by 7-hydroxystaurosporine, staurosporine, flavopyridol, CYC202 (Cyclacel) and BMS-387032.

"Cell proliferation inhibitors and survival signaling pathway" refers to pharmaceutical agents that inhibit cell surface receptors and signal transduction cascades downstream of those surface receptors. Such agents include inhibitors of EGFR inhibitors (eg gefitinib and erlotinib), ERB-2 inhibitors (for example trastuzumab), IGFR inhibitors (for example those disclosed in WO 03/059951), cytokine receptor inhibitors, MET inhibitors, PI3K inhibitors (for example LY294002), serine / threonine kinases (including but not limited to Akt inhibitors such as those described in the documents (WO 03/086404, WO 03/086403, WO 03/086394, WO 03 / 086279; WO 02/083675, WO 02/083139, WO 02/083140 and WO 02/083138), Raf kinase inhibitors (for example BAY-43-9006), MEK inhibitors (for example CI-1040 and PD- 098059) and mTOR inhibitors (eg Wyeth CCI-779 and Ariad AP23573) Such agents include small molecule inhibitor compounds and antibody antagonists.

"Apoptosis inducing agents" include activators of members of the TNF receptor family (including TRAIL receptors).

In one embodiment the compounds of the present invention are useful for treating cancer in combination with one or more, particularly one, two or three agents selected from temozolomide, cisplatin, carboplatin, oxaliplatin, trinotecan and topotecan.

A compound of the present invention may also be useful for treating cancer in combination with any one.

or more of the following therapeutic agents: abarelix (Plenaxis depot®); aldesleukin (Prokine®); Aldesleukin (Proleukin®); Alemtuzumab (Campath®); alitretinoin (Panretin®); allopurinol (Zyloprim®); altretamine (Hexalen®); amifostine (Ethy-ol®); anastrozole (Arimidex®); arsenic trioxide (Trisenox®); asparaginase (Elspar®); azacitidine (Vidaza®); bevacuzimab (Avastin®); Bexarotene capsules (Targretin®); Bexarotene gel (Targretin®); bleomycin (Blenoxane®); bortezomib (Vel-cade®); intravenous busulfan (Busulfex®); oral busulfan (Myleran®); calusterone (Methosarb®); Capecitabine (Xeloda®); carboplatin (Paraplatin®); carmustine (BCNU®, BiCNU®); carmustine (Gliadel®); carmustine with Polifeprosan20 implant (Gliadel Wafer®); celecoxib (Celebrex®); cetuximab (Erbitux®); chlorambucil (Leukeran®); cisplatin (Platinol®); cladribine (Leustatin®, 2-CdA®); clofarabine (Clolar®); cyclophosphamide (Cytoxan®, Neosar®); cyclophosphamide (Cytoxan Injection®); cyclophosphamide (CytoxanTablet®); cytarabine (Cytosar-U®); liposomal cytarabine (DepoCyt®); dacarbacin (DTIG-Dome®); dactinomycin, actinomycin D (Cosmegen®); Darbepoetin alfa (Aranesp®); liposomal daunorubicin (DanuoXome®); daunorubicin, daunomycin (Daunorubicin®); daunorubicin, daunomycin (Cerubidine®); Denileucine diftitox (Ontak®); dexrazoxane (Zinecard®); docetaxel (Taxotere®); doxorubicin (Adriamycin PFS®); doxorubicin (Adriamycin®, Rubex®); doxorubicin (Adriamycin PFS Injection®); liposomal doxorubicin (Doxil®); dromostanolone propionate (Dromostanolone®); dromostanolone propionate (Masterone Injection®); Elliott Solution B (Elliott’s B Solution®); epirubicin (Ellence®); Epoetin alfa (epogen®); Erlotinib (Tarceva®); estramustine (Emcyt®); Etoposide phosphate (Etopophos®); etoposide, VP-16 (Vepesid®); exemestane (Aromasin®); Filgrastim (Neupogen®); floxuridine (intraarterial) (FUDR®); fludarabine (Fludara®); fluorouracil, 5-FU (Adrucil®); fulvestrant (Faslodex®); gefitinib (Iressa®); gemcitabine (Gemzar®); gemtuzumab ozogamycin (Mylotarg®); goserelin acetate (Zoladex Implant®); goserelin acetate (Zoladex®); histrelin acetate (Histrelin implant®); hydroxyurea (Hydrea®); Ibritumomab tiuxetane (Zevalin®); idarubicin (Idamycin®); ifosfamide (IFEX®); Imatinib Mesylate (Gleevec®); interferon alfa 2a (Roferon A®); Interferon alfa-2b (Intron A®); irinotecan (Camptosar®); lenalidomide (Revlimid®); letrozole (Femara®); leucovorin (Wellcovorin®, Leucovorin®); Leuprolide Acetate (Eligard®); levamisole (Ergamisol®); lomustine, CCNU (CeeBU®); mechlorethamine, hydrogen mustard (Mustargen®); megestrol acetate (Megace®); melfalan, L-PAM (Alkeran®); mercaptopurine, 6-MP (Purinethol®); mesna (Mesnex®); mesna (Mesnex tabs®); methotrexate (Methotrexate®); methoxsalene (Uvadex®); mitomycin C (Mutamycin®); mitotane (Lysodren®); mitoxantrone (Novantrone®); Nandrolone Phepropionate (Durabolin-50®); nelarabine (Arranon®); Nofetumomab (Verluma®); Oprelvequina (Neumega®); oxaliplatin (Eloxatin®); paclitaxel (Paxene®); paclitaxel (Taxol®); Paclitaxel protein bound particles (Abraxane®); palifermina (Kepivance®); pamidronate (Aredia®); pegademasa (Adagen (Pegademase Bovine) ®); pegaspargasa (Oncaspar®); Pegfilgrastim (Neulasta®); pemetrexed disodium (Alimta®); pentostatin (Nipent®); pipobroman (Vercyte®); plicamycin, mitramycin (Mithracin®); porfimer sodium (Photofrin®); procarbacin (Maculane®); quinacrine (Atabrine®); Rasburicase (Elitek®); Rituximab (Rituxan®); sargramostim (Leukine®); Sargramostim (Prokine®); sorafenib (Nexavar®); streptozocin (Zanosar®); sunitinib maleate (Sutent®); talc (Sclerosol®); tamoxifen (Nolvadex®); temozolomide (Temodar®); teniposide, VM-26 (Vumon®); testolactone (Teslac®); thioguanine, 6-TG (Thioguanine®); thiotepa (Thioplex®); topotecan (Hycamtin®); toremifene (Fareston®); Tositumomab (Bexxar®); Tositumomab / I-131 tositumomab (Bexxar®); Trastuzumab (Herceptin®); Tretinoin, ATRA (Vesanoid®); Uracil mustard (Uracil Mustard Capsules®); Valrubicin (Valstar®); vinblastine (Velban®); vincristine (Oncovin®); vinorelbine (Navelbine®); vorinostat (Zolinza®); zoledronate (Zometa®); nilotinib (Tasigna®) and dasatinib (Sprycel®).

The invention also encompasses combinations with NSAIDs that are selective COX-2 inhibitors. For the purposes of this specification, NSAIDs that are selective COX-2 inhibitors are defined as those that have a specificity to inhibit COX-2 over COX-1 at least 100 times as measured by the ratio of IC50 to COX -2 vs. IC50 for COX-1 evaluated by microsomal or cellular assays. Such compounds include, but are not limited to those disclosed in United States Patent 5,474,995, United States Patent 5,861,419, United States Patent 6,001,843, United States Patent 6,020,343, United States Patent 5,409,944 , U.S. Patent 5,436,265, U.S. Patent 5,536,752, U.S. Patent 5,550,142, U.S. Patent 5,604,260, US Document 5,698,584, U.S. Patent 5,710,140, WO 94/15932, U.S. Patent 5,344,991, U.S. Patent 5,134,142, U.S. Patent 5,380,738, U.S. Patent 5,393,790, U.S. Patent 5,466,823, U.S. Patent 5,633,272 and U.S. Patent 5,932,598, all of which are hereby incorporated by reference.

COX-2 inhibitors that are particularly useful in the present treatment process are 5-chloro-3 (4-methylsulfonyl) phenyl-2- (2-methyl-5-pyridinyl) pyridine; or a pharmaceutically acceptable salt thereof.

Compounds that have been described as specific COX-2 inhibitors and are therefore useful in the present invention include, but are not limited to: parecoxib, CELEBREX® and BEXTRA® or a pharmaceutically acceptable salt thereof.

Other examples of angiogenesis inhibitors include, but are not limited to, endostatin, ukraine, ranpyrnase, IM862, 5-methoxy-4- [2-methyl-3- (3-methyl-2-butenyl) oxyranyl] -1-oxaspiro [2 , 5] oct-6-yl (chloroacetyl) carbamate, acetyldinanaline, 5 amino-1 - [[3,5-dichloro-4- (4-chlorobenzoyl) phenyl] methyl] -1H-1,2,3-triazol-4 -carboxamide, CM101, squalamin, combretastatin, RPI4610, NX31838, sulfated manopentaose phosphate, 7.7- (carbonyl-bis [imino-N-methyl-4,2-pyrrolocarbonylimino [N-methyl-4,2-pyrrole] -carbonylimino] -bis- (1,3-naphthalene disulfonate) and 3 - [(2,4-dimethylpyrrol-5-yl) methylene] 2-indolinone (SU5416).

As previously used, "integrin blockers" refers to compounds that selectively antagonize, inhibit or counteract the binding of a physiological ligand to the αvβ3 integrin, to compounds that selectively antagonize, inhibit or counteract the binding of a physiological ligand to the integrin αvβ5 to compounds that antagonize, inhibit or counteract the binding of a physiological ligand to both αvβ3 integrin and αvβ5 integrin and to compounds that antagonize, inhibit or counteract the activity of particular integrin or integrins expressed in capillary endothelial cells. The term also refers to antagonists of integrins αvβ6, αvβ8 α1β1, α2β1, α5β1, α6β1 and α6β4. The term also refers to antagonists of any combination of integrins αvβ3, αvβ5, αvβ6, αvβ8, α1β1, α2β1, α5β1, α6β1, and α6β4.

Some specific examples of tyrosine kinase inhibitors include N- (trifluoromethylphenyl) -5-methylisoxazol-4carboxamide, 3 - [(2,4-dimethylpyrrol-5-yl) methylidenyl) indolin-2-one, 17- (allylamino) -17 -demethoxyigeldanamicin, 4- (3-chloro-4-fluorophenylamino) -7-methoxy-6- [3- (4-morpholinyl) propoxy] quinazoline, N- (3-ethynylphenyl) -6,7-bis (2-methoxyethoxy) - 4quinazolinamine, BIBX1382, 2,3,9,10,11,12-hexahydro-10- (hydroxymethyl) -10-hydroxy-9-methyl-9,12-epoxy-1 Hdiindole [1,2,3-fg: 3 ' , 2 ', 1'-kl] pyrrolo [3,4-i] [1,6] benzodiazocin-1-one, SH268, genistein, STI571, CEP2563, 4- (3-chlorophenylamino) -5,6-dimethyl-7H- pyrrolo [2,3-d] pyrimidinmethane sulfonate, 4- (3-bromo-4-hydroxyphenyl) amino-6,7-dimethoxyquinazoline, 4- (4'-hydroxyphenyl) amino-6,7-dimethoxyquinazoline, SU6668, STI571A, N- 4-chlorophenyl-4- (4-pyridylmethyl) -1-phthalacinamine and EMD121974.

In one embodiment, the compounds of the present invention are useful for the treatment or prevention of the onset of necrosis induced by selective N3-adenine methylating agents such as MeOSO2 (CH2) lexitropsin (Me-Lex).

Combinations with compounds other than antineoplastic compounds are also encompassed in the present processes. For example, combinations of the compounds presently claimed with PPAR-γ agonists (i.e., PPAR-gamma) and PPAR-δ agonists (i.e., PPAR-delta) are useful in the treatment of certain malignant tumors. PPAR-γ and PPAR-δ are proliferative activated receptors for nuclear peroxisome γ and δ. The expression of PPAR-γ in endothelial cells and its involvement in angiogenesis has been indicated in the literature (see J. Cardiovasc. Pharmacol. (1998) 31: 909-913; J. Biol. Chem. (1999) 274: 9116- 9121; Invest. Ophthalmol Vis. Sci. (2000) 41: 2309-2317). More recently, PPAR-γ agonists have been shown to inhibit the angiogenic response to VEGF in vitro; Both rosiglitazone and troglitazone maleate inhibit the development of retinal neovascularization in mice. (Arch. Ophthamol. (2001) 119: 709-717). Examples of PPAR-γ agonists and PPAR-γ / α agonists include, but are not limited to, thiazolidinediones (such as DRF2725, CS-011, troglitazone, rosiglitazone and pioglitazone), fenofibrate, gemfibrocil, clofibrate, GW2570, SB219994 -H039242, JTT-501, MCC-555, GW2331, GW409544, NN2344, KRP297, NP0110, DRF4158, NN622, GI262570, PNU182716, DRF552926, acid 2 - [(5,7-dipropyl-3-trifluoromethyl bencisoxazol-6-yl) oxy] -2-methylpropionic (disclosed in USSN 09 / 782,856) and 2 (R) -7- (3- (2-chloro-4- (4-fluorophenoxy) phenoxy) propoxy) -2-ethylchroman-2-carboxylic (disclosed in USSN 60 / 235,708 and 60 / 244,697).

Another embodiment of the present invention is the use of the presently disclosed compounds in combination with antiviral agents (such as nucleoside analogs including ganciclovir for cancer treatment. See WO 98/04290.

Another embodiment of the present invention is the use of the presently disclosed compounds in combination with gene therapy for the treatment of cancer. For an overview of genetic strategies to treat cancer see Hall et al (Am J Hum Genet (1997) 61: 785-789) and Kufe et al (Cancer Medicine, 5th Ed, p. 876-889, BC Decker, Hamilton 2000). Gene therapy can be used to deliver any tumor suppressor gene. Examples of such genes include, but are not limited to, p53, which can be delivered by recombinant virus-mediated gene transfer (see US Patent No. 6,069,134, for example), a uPA / uPAR antagonist ("Adenovirus-Mediated Delivery of a uPA / uPAR Antagonist Suppresses Angiogenesis-Dependent Tumor Growth and Dissemination in Mice, ”Gene Therapy, August (1998) 5 (8): 1105-13) and gamma interferon (J Immunol (2000) 164: 217-222).

The compounds of the present invention can also be administered in combination with an inherent multi-drug resistance (MDR) inhibitor, in particular MDR associated with high levels of transporter protein expression. Such MDR inhibitors include p-glycoprotein (P-gp) inhibitors, such as LY335979, XR9576, OC144-093, R101922, VX853, verapamil and PSC833 (waltzpodar).

A compound of the present invention may be used together with anti-emetic agents to treat nausea or emesis, including acute, delayed, late and anticipatory emesis, which may result from the use of a compound of the present invention, alone or with radiation therapy. For the prevention or treatment of emesis, a compound of the present invention can be used together with other anti-emetic agents, especially neuroquinine 1 receptor antagonists, 5HT3 receptor antagonists, such as ondansetron, granisetron, tropisetron and zathisetron, GABAB receptor agonists. , such as baclofen, a corticosteroid such as Decadron (dexamethasone), Kenalog, Aristocort, Nasalide, Preferid, Benecorten and others such as those disclosed in U.S. Patents No. 2,789,118, 2,990,401, 3,048,581, 3,126. 375, 3,929,768, 3,996,359, 3,928,326 and 3,749,712, an antidopaminergic, such as phenothiazines (e.g., prochlorperazine, flufenacin, thioridacin and mesoridacin), metoclopramide or dronabinol. In one embodiment, an anti-emesis agent selected from a neurokinin 1 receptor antagonist, a 5HT3 receptor antagonist and a corticosteroid is administered as an adjuvant for the treatment or prevention of emesis that may result after administration of the present compounds.

Neurokinin-1 receptor antagonists of use in conjunction with the compounds of the present invention are fully described, for example, in U.S. Patent Nos. 5,162,339, 5,232,929, 5,242,930, 5,373,003, 5,387. 595, 5,459,270, 5,494,926, 5,496,833, 5,637,699, 5,719,147; European Patent Publications No. EP 0 360 390, 0 394 989, 0 428 434, 0 429 366, 0 430 771, 0 436 334, 0 443 132, 0 482 539, 0 498 069, 0 499 313, 0 512 901, 0 512 902, 0 514 273, 0 514 274, 0 514 275, 0 514 276, 0 515 681, 0 517 589, 0 520 555, 0 522 808, 0 528 495, 0 532 456, 0 533 280, 0 536 817, 0 545 478, 0 558 156, 0 577 394, 0 585 913.0 590 152, 0 599 538, 0 610 793, 0 634 402, 0 686 629, 0 693 489, 0 694 535, 0 699 655, 0 699 674, 0 707 006, 0 708 101, 0 709 375, 0 709 376, 0 714 891, 0 723 959, 0 733 632 and 0 776 893; PCT International Patent Publications No. WO 90/05525, 90/05729, 91/09844, 91/18899, 92/01688, 92/06079, 92/12151, 92/15585, 92/17449, 92/20661, 92/20676 , 92/21677, 92/22569, 93/00330, 93/00331, 93/01159, 93/01165, 93/01169, 93/01170, 93/06099, 93/09116, 93/10073, 93/14084, 93 / 14113, 93/18023, 93/19064, 93/21155, 93/21181, 93/23380, 93/24465, 94/00440, 94/01402, 94/02461, 94/02595, 94/03429, 94/03445 , 94/04494, 94/04496, 94/05625, 94/07843, 94/08997, 94/10165, 94/10167, 94/10168, 94/10170, 94/11368, 94/13639, 94/13663, 94 / 14767, 94/15903, 94/19320, 94/19323, 94/20500, 94/26735, 94/26740, 94/29309, 95/02595, 95/04040, 95/04042, 95/06645, 95/07886 , 95/07908, 95/08549, 95/11880, 95/14017, 95/15311, 95/16679, 95/17382, 95/18124, 95/18129, 95/19344, 95/20575, 95/21819, 95 / 22525, 95/23798, 95/26338, 95/28418, 95/30674, 95/30687, 95/33744, 96/05181, 96/05193, 96/05203, 96/06094, 96/07649, 96/10562 , 96/16939, 96/18643, 96/20197, 96/21661, 96/29304, 96/29317, 96/29326, 96/29328, 96/31214, 96/32385, 96/37489, 97/01553, 97/01554, 97/03066, 97/08144; 97/14671, 97/17362, 97/18206, 97/19084, 97/19942 and 97/21702; and in British Patent Publications No. 2 266 529, 2 268 931, 2 269 170, 2 269 590, 2 271 774, 2 292 144, 2 293 168, 2 293 169 and 2 302 689. The preparation of such compounds is fully described in the aforementioned patents and publications, which are incorporated herein by reference.

In one embodiment, the neuroquinine 1 receptor antagonist for use in conjunction with the compounds of the present invention are selected from: 2- (R) - (1- (R) - (3,5-bis (trifluoromethyl) phenyl) ethoxy) -3- (S) - (4-fluorophenyl) -4- (3- (5-oxo1H, 4H-1,2,4-triazolo) methyl) morpholine or a pharmaceutically acceptable salt thereof, which is described in U.S. Patent No. 5,719,147.

A compound of the present invention can also be administered with an agent useful in the treatment of anemia. Such an anemia treatment agent is, for example, a continuous erythropoiesis receptor activator (such as epoetin alfa).

A compound of the present invention can also be administered with an agent useful in the treatment of neutropenia. Said neutropenia treatment agent is, for example, a hematopoietic growth factor that regulates the production and function of neutrophils such as a human granulocyte colony stimulator factor, (G-CSF). Examples of a G-CSF include filgrastim.

A compound of the present invention can also be administered with an immunological enhancer drug, such as levamisole, isoprinosine and Zadaxin.

A compound of the present invention may also be useful for the treatment or prevention of cancer, including bone cancer, in combination with bisphosphonates (which is understood to include bisphosphonates, diphosphonates, bisphosphonic acids and diphosphonic acids). Examples of bisphosphonates include but are not limited to: etidronate (Didronel), pamidronate (Aredia), alendronate (Fosamax), risedronate (Actonel), zoledronate (Zometa), ibandronate (Boniva), incadronate or cimadronate, clodronate, EB-1053, minodronate , neridronate, pyridronate and tiludronate including any and all pharmaceutically acceptable salts, derivatives, hydrates and mixtures thereof.

Therefore, the scope of the present invention encompasses the compounds presently claimed for use in combination with ionizing radiation and / or in combination with a second compound selected from: HDAC inhibitors, an estrogen receptor modulator, a receptor modulator of androgens, a retinoid receptor modulator, a cytotoxic / cytostatic agent, an antiproliferative agent, a prenyl protein transferase inhibitor, an HMG-CoA reductase inhibitor, an angiogenesis inhibitor, a PPAR-γ agonist, an agonist PPAR-δ, an antiviral agent, an inherent multi-drug resistance inhibitor, an antiemetic agent, an agent useful in the treatment of anemia, an agent useful in the treatment of neutropenia, an immunological enhancer drug, a proliferation signaling inhibitor and cell survival, an agent that interferes with a cell cycle control point, an apoptosis inducing agent and a bisphosphorus born.

The term "administration" and variants thereof (eg, "administering" a compound) in reference to a compound of the invention means introducing the compound or a prodrug of the compound into the animal's system that needs treatment. When a compound of the invention or prodrug thereof is provided in combination with one or more additional active agents (for example, a cytotoxic agent, etc.), it is understood that "administration" and its variants each include simultaneous and sequential introduction of the compound or prodrug thereof and other agents.

As used herein, it is intended that the term "composition" encompasses a product comprising the specified ingredients in the specified amounts, as well as any product that results, directly or indirectly, from the combination of the specified ingredients and the amounts specified.

The term "therapeutically effective amount" as used herein means the amount of active compound or pharmaceutical agent that induces the biological or medicinal response in a tissue, system, animal.

or human being that is sought by a researcher, veterinarian, medical doctor or other clinical specialist.

The term "treatment" refers to the treatment of a mammal afflicted with a pathological condition and refers to an effect that alleviates the condition by killing the cancer cells, but also to an effect that results in the inhibition of the progress of the condition and includes a reduction in the speed of progress, a stop in the speed of progress, relief of the condition and cure of the condition. Treatment is also included as a prophylactic measure (ie prophylaxis).

The term "pharmaceutically acceptable" as used herein refers to compounds, materials, compositions and / or pharmaceutical forms that are, within the scope of reasonable medical criteria, suitable for use in contact with the tissues of a subject ( for example, being human) without excessive toxicity, irritation, allergic response or other problem or complication, adequate with a reasonable benefit / risk ratio. Each vehicle, excipient, etc. It must also be “acceptable” in the sense of being compatible with the other ingredients of the formulation.

The term "accessory" refers to the use of compounds together with known therapeutic means. Such means include cytotoxic drug regimens and / or ionizing radiation as used in the treatment of different types of cancer. In particular, it is known that the active compounds potentiate the actions of various chemotherapeutic cancer treatments, which include the class of topoisomerase poisons (for example topotecan, irinotecan, rubitecan), most of the known alkylating agents (for example DTIC, temozolamide ) and platinum-based drugs (for example, carboplatin, cisplatin) used in the treatment of cancer.

These and other aspects of the invention will be apparent from the teachings contained herein.

They are abbreviations based on the description of the chemistry and the following examples: AcCl (acetyl chloride); (BzO) 2 (benzoyl peroxide); Cbz-Cl (benzylchloroformate); DCM (dichloromethane); DIPEA (di-iso-propylethylamine); DMF (dimethylformamide); DMSO (dimethyl sulfoxide); eq. (equivalent); ES (electrospray); EtOAc (ethyl acetate); EtOH (ethanol); sieve mol. (molecular sieves); HATU [O- (7-azabenzotriazol-1-yl) -N, N, N ’, N’tetramethyluronium hexafluoro-phosphate]; MeCN (acetonitrile); MeOH (methanol); MS (mass spectrometry); MW (microwave); NBS (N-bromosuccinimide); NMMO (N-methylmorpholin-N-oxide); NMR (nuclear magnetic resonance); Pcol (column pressure); iPrOH (isopropanol); TA (room temperature); ac. sat. (saturated aqueous); SiO2 (silica gel); and THF (tetrahydrofuran). t-BuOH (tert-butanol); KOAc (potassium acetate); MW microwave; SPE IST ISOLUTE® column SCX (cation exchange resin of International Sorbent Technology ISOLUTE® Solid Phase Extraction column); CFS (critical / liquid phase chromatography); TBTU O- (1H benzotriazol-1-yl) N, N, N ’, N’-tetramethyluronium tetrafluoroborate; and Tcol (column temperature). CDCl3 (deuterium chloroform); TLC (thin layer chromatography) and TFA (trifluoroacetic acid).

Compounds of formula I can be prepared by reacting a compound of formula IA with ammonia:

image 1

wherein R1 and R2 are as defined above and Rx is C1-6 alkyl, such as methyl. The reaction is generally carried out using an aqueous solution of NH3 in a solvent, such as THF at about 70 ° C, in a sealed reaction vessel (with caution). Alternatively, a base such as NaOH or KOH 5 can be added to hydrolyze the ester to give the corresponding carboxylic acid (Rx is hydrogen) followed by the addition of NH3 in the presence of coupling agent, such as HATU or TBTU and DIPEA in a solvent such as DMF, the reaction being carried out at approximately room temperature. Alternatively, the carboxylic acid can be activated to form a mixed anhydride, for example using Boc2O, and then reacted with ammonium bicarbonate, generally in a solvent such as pyridine. Alternatively, the ester can become

10 compounds of formula IA using ammonia in a solvent, such as MeOH at about 120 ° C, for example in a microwave.

The nitrogen atom in the piperidine ring in the compounds of formula IA can be protected during the above synthesis, for example by Boc.

Compounds of formula IA can be prepared by reacting a compound of formula IB with an azide:

fifteen

image 1

in which R1, R2 and Rx are as defined above. An azide, such as NaN3, can be used generally in a solvent such as DMF of about 90 ° C to 140 ° C. An additive, such as 2,6-lutidine, can also be used. The reaction can be carried out under a nitrogen atmosphere.

Compounds of formula IB may be prepared by the condensation of a compound of formula IC with a compound of formula ID:

image 1

wherein R1, R2 and Rx are as defined above and L1 is a leaving group, such as nitro or halogen, for example fluorine. The processes include condensation in the presence of a dehydrogenation agent, such as MgSO4 or molecular sieves, or heating in an alcohol solvent, such as reflux ethanol. The

The reaction can be carried out in a nitrogen atmosphere.

Compounds of formula IC can be prepared by oxidizing a compound of formula IE with an oxidizing agent, such as NMMO:

image 1

wherein R1, Rx and L1 are as defined above and L2 is a leaving group, such as halogen, for example bromine, generally in a solvent such as MeCN at about room temperature. The reaction can be carried out under a nitrogen atmosphere.

Compounds of formula IE, in which L2 is bromine can be prepared by oxidizing a compound of formula IF with a brominating agent, such as NBS in the presence of a radical initiator, such as benzoyl peroxide:

image 1

In which R1, Rx and L1 are as defined above, generally in a solvent such as CCl4 under reflux. The reaction can be carried out under a nitrogen atmosphere.

Compounds of formula IF, in which L1 is fluorine can be prepared by diazotizing a compound of formula IG:

image 1

In which R1 and Rx are as defined above followed by the decomposition of the diazonium salt of the intermediate. For example, diazotization can be performed using nitrosium tetrafluoroborate in a solvent such as DCM at about 0 ° C. Then, the corresponding diazonium tetrafluoroborate salt can be isolated and subsequently decomposed at elevated temperatures to give the corresponding fluorobenzene derivative (Caution), such as by heating at 160 ° C in a solvent, such as dichlorobenzene.

The compounds of formula IF, wherein L1 is nitro can be prepared by nitration of a compound of formula 1H:

image 1

wherein R1 is as defined above followed by esterification. The nitration reaction can

be performed in the presence of a nitrate, such as potassium nitrate and an acid, such as sulfuric acid a

about room temperature The esterification stage can be carried out under conditions

5 conventional, such as by reaction with an alkyl halide of formula Rx-X, wherein X is a halogen, such

as iodine, in the presence of a base, such as cesium carbonate, and in a solvent, such as DMF a

about room temperature An alcohol of the formula Rx-OH may also be used together with a

acid catalyst, such as HCl generated in situ from AcCl / MeOH, at reflux. Afterwards, the

compound of formula IF desired by hydrogenation of the nitro compound, giving the corresponding aniline using hydrogen and a catalyst, such as palladium on carbon, typically in an alcoholic solvent, such as

MeOH

Alternatively, compounds of formula I can be prepared by reducing a compound of formula IJ:

image 1

in which R1 and R2 are as defined above. The reduction can be performed in a Fowler 15 reaction using an acyl chloride, such as CBz-Cl and a reducing agent, such as NaBH4. Hydrogenation on palladium on carbon completes the reaction and eliminates the CBz protecting group.

Compounds of formula IJ can be prepared by cross-linking a compound of formula IK with 3-pyridinylboronic acid of formula IL:

image 1

20 in which R1, R2 and L2 are as defined above. The reaction is generally carried out under Suzuki coupling conditions, such as using catalysts, such as Pd2 (dba) 3 and tri (tert-butyl) phosphine together with a base, such as sodium carbonate and solvents, such as DMF and water at approximately 90 ° C.

Compounds of formula IK can be prepared by condensation of a compound of formula IM with a compound of formula IN:

image 1

5

10

fifteen

twenty

25

30

35

wherein R1, R2 and L2 are as defined above and L3 is a leaving group such as halogen, for example fluorine, generally in a solvent, such as DMF at about 180 ° C in a microwave. A base, such as K2CO3, can also be added.

Compounds of formula IM can be prepared by reacting a compound of formula IO:

image 1

wherein R1 and Rx are as defined above, with a base such as KOH or NaOH at about room temperature to hydrolyze the ester to give the corresponding carboxylic acid (Rx is hydrogen) followed by the addition of NH3 in the presence of coupling agents, such as HATU, DIPEA and TBTU in a solvent, such as DMF, the reaction being carried out at approximately room temperature.

The compounds of formula IO can be prepared from the compound of formula IG by acetylation of the aniline group with reagents such as acetyl chloride in a solvent, such as 1,2-DCE at about 55 ° C. Then, the cyclization to give the desired indazole can be carried out by treatment with sodium nitrite in acid, for example concentrated hydrochloric acid, generally in the presence of a co-solvent, such as toluene and water at about 0 ° C.

When the synthesis of intermediates and starting materials is not described, these compounds are commercially available or can be manufactured from commercially available compounds by conventional procedures or by extension of the synthesis described above, schemes and Examples in This document.

The compounds of formula I can be converted into other compounds of formula I by known procedures or by procedures described in the above synthesis, schemes and Examples herein.

During any of the synthetic sequences described herein, it may be necessary and / or desirable to protect sensitive or reactive groups in any of the molecules of interest. This can be achieved by means of conventional protecting groups, such as those described in, Protecting Groups in Organic Synthesis, 3rd Edition, Greene, T. W. and Wuts, P. G. M .; Wiley Interscience, 1999 and Kocienski, P. J. Protecting Groups, Thieme, 1994. Protective groups can be removed at a convenient later stage using methods known in the art. For example, when the Boc (tert-butoxycarbonyl) or benzylcarbonyl protecting group is present, it can be removed by the addition of solvents such as, TFA, DCM and / or MeCN at about room temperature. The compound can also be hydrogenated using conventional procedures, such as treatment with a catalyst, such as Pd / C, in a solvent such as methanol in a hydrogen atmosphere. EtOAc can also be added in the presence of HCl and 1,4-dioxane to remove the Boc or benzylcarbonyl protecting group, at about room temperature.

The compounds of this invention were prepared according to the following schemes. All the variables in the formulas are as defined above.

When the compounds of the present invention have chiral centers, the enantiomers can be separated in the racemic mixtures by conventional separation procedures, such as using SFC, chiral HPLC or chiral acid resolution. The separation can be carried out at any stage of the process by manufacturing the compounds of formula I. Therefore, the separation can be carried out in the final stage, or alternatively, the intermediates can be separated and then used particular enantiomers in subsequent reactions to produce the desired products. .

5 Scheme 1

A procedure for synthesizing derivatives of the compounds of this invention is shown in Scheme 1, in this way the substituted 2H-indazoles are prepared using a synthetic route similar to that described in WO 2005/066136. Upon initial conversion of the 2-nitro-3-methylbenzoic acid derivative into the corresponding ester, radical bromination of the methyl group using reagents such as N-bromosuccinimide and benzoyl peroxide 10 produces the key benzyl bromide derivative. The oxidation of this benzyl bromide to give the corresponding benzaldehyde can be carried out, for example, using N-methylmorpholine N-oxide and molecular sieves. After condensation of the aldehyde with an amine, the ring closure can be performed by treatment by treating the key intermediate with sodium azide at elevated temperature to introduce the final nitrogen atom and the resulting nitrogen extrusion to provide the indazole ring. A base can also be added, such as

15 lutidine to this reaction. The final conversion of ester into the primary amide produces the desired derivatives. This can be done by heating the ester in an ammonia solution or by conversion to the corresponding carboxylic acid and then by amide coupling.

image 1

Scheme 1

20 Scheme 2

Below is a variation of scheme 1 in scheme 2 and allows the introduction of substituents into indazole nuclei. When the required nitrobenzoic acid derivatives are not commercially available, they can be prepared through nitration of the corresponding benzoic acid derivatives, for example, using potassium nitrate in concentrated sulfuric acid. Synthetic manipulations as described above allow the formation of aniline that can be cyclized to give indazole by first acetylation of indazole and cyclisation with sodium nitrite in concentrated acid HCl at 0 ° C. Alternatively, the aniline can be diazotized with nitrosium tetrafluoroborate and diazonium tetrafluoroborate salt decomposed at elevated temperatures to give the corresponding dilfluorobenzene derivative by a Schiemann reaction (Caution). Then the synthetic sequence as described in scheme 1 allows the oxidation of the group

methyl benzyl to give the corresponding aldehyde and the elaboration of the desired indazole derivatives by coupling with a (hetero) anilide and cyclisation with sodium azide.

image 1

Scheme 2

5 Scheme 3

An alternative procedure involves the functionalization of indazole at a later stage as shown in scheme 3. Here, the indazole ester is first converted into the corresponding carboxamide and subjected to nucleophilic aromatic substitution of the aromatic (hetero) aromatic fluoro bromide . This allows the preparation of a bromide derivative that can be coupled under Suzuki coupling conditions, for example, using tri (tert.

10 butyl) phosphine and Pd2 (dba) 3 as a catalyst in the presence of a base, such as sadic carbonate. Then, the conversion to the desired piperidine moiety is carried out by a Fowler reaction using an acyl chloride, such as CBz-Cl and a reducing agent, such as NaBH4. The final hydrogenation reaction can produce the corresponding piperidine derivatives.

image 1

Scheme 3

PARP-1 SPA test

The exemplified compounds described herein were tested in this assay and found to have an IC50 value of less than 5 µM, particularly less than 50 M.

Work Reagents

Assay buffer: 100 mM Tris pH 8, 4 mM MgCl2, 4 mM spermine, 200 mM KCI, 0.04% Nonidet P-40.

Enzyme Blend: Test buffer (12.5 µl), 100 mM DTT (0.5 µl), PARP-1 (5 nM, Trevigen 4668-500-01), H2O (up to 35 µl).

Nicotinamide adenine dinucleotide (NAD) / DNA mixture: [3H-NAD] (250 uCi / ml, 0.4 µl, Perkin-Elmer NET443H), NAD (1.5 mM, 0.05 µl, SIGMA N-1511 ), Biotinylated NAD (250 µM, 0.03 µl, Trevigen 4670-500-01), activated calf thymus (1 mg / ml, 0.05 µl, Amersham Biosciences 27-4575), H2O (up to 10 µl).

Development mixture: Streptavidin SPA beads (5 mg / ml, Amersham Biosciences RPNQ 0007) dissolved in 500 mM EDTA.

Experimental design

The reaction is carried out in a 96-well microplate with a final volume of 50 µl / well. Add 5 µl of 5% DMSO / compound solution, add enzyme mixture (35 µl), start the reaction by adding NAD / DNA mixture (10 µl) and incubate for 2 hours at RT. Stop the reaction by adding development mixture (25 µl) and incubate 15 minutes at RT. Measure using a Packard TOP COUNT instrument.

Proliferation assay in silenced HeLa cells for BRCA-1. Abbreviations:

IMDM (Dulbecco Medium Modified by Iscove); RPMI (Medi Roswell Park Memorial Institute); MOI (multiplicity of infection); GFP (green fluorescent protein); PBS (Phosphate Buffered Saline); FCS (fetal calf serum); and DMEM (Eagle Medium Modified by Dulbecco).

The compounds of the present invention were also tested in an antiproliferative assay in HeLa BRCA1wt and BRCA1- (shRNA) cells in matching pairs. The assay shows that PARP inhibitors are capable of showing selectivity with growth inhibition of BRCA-deficient cells. The compounds showed CC50 less than 5 µM in BRCA1 deficient cells and a selectivity greater than 10 times over the BRCA competent cells.

The assay is based on the ability of living cells to convert a redox dye (resazurine) into a fluorescent final product (resofurin). The amount of resofurin produced is directly proportional to the number of cells.

Cell lines:

HeLa shBRCA1-GFP - These are HeLa cells transduced to an MOI of 100 with a Lentivirus that contains an mRNA against BRCA-1 and an expression cassette for GFP. BRCA-1 silencing is greater than 80% as assessed by Taqman analysis and cells stably express GFP.

HeLa THM-GFP - These are HeLa cells transduced to an MOI of 100 with a control vector that does not express any mRNA.

Protocol

-Seed 300 cells / well in 96-well black plate in 90µl of Culture Medium *: -Include 4 hours at 37 ºC, CO2 5%

-Add 10 µl / well of the 10X compound (5% DMSO in H2O) -Incubate for 168 hours at 37 ° C, 5% CO2 -Add 10 µl of Celltiter Blue solution (Promega, G8081) prediluted 1: 1 in PBS1x -Include the mix for 45 'at 37 ° C, 5% CO2 -Incubate 15' at RT in the dark -Read the plate on the fluorimeter ex: 550 nm; em: 590 nm * Culture Medium: DMEM (GIBCO, 41966-029), FCS 10% (GIBCO, 10106-169), Penicillin-Streptomycin 0.1 mg / ml

(GIB-CO, 15140-114), 2mM L-Glutamine (GIBCO, 3042190)

Proliferation assay in BRCA-deficient cell lines naturally.

It was also shown that the compounds of the present invention inhibit the proliferation of cell lines deficient in BRCA-1 (MDA-MB-436) and BRCA-2 (CAPAN-1) naturally with CC50 less than 5 micromolar.

Proliferation Assay

The cells are seeded in a 96-well plate at 700 cells / well in 100 µl of the appropriate medium / well *. The next day, serial dilutions of the compound are added in a final volume of 200 µl / well. Each dilution is tested in triplicate.

Six days later, cell viability is estimated using CellTiter-Blue Cell Viability Test according to the manufacturer's instructions (Promega). The plates are read in the Fusion Alpha microplate reader (Packard Bioscience).

For low proliferation cell lines (ie CAPAN-1), proliferation is tested 14 days after adding the compounds and changing the medium once on day 7 (170 µl of medium is aspirated per well and replaced with 170 µl of fresh medium containing the compounds).

* Culture Medium: MDA-MB-436: RPMI (GIBCO), FBS 10% (CO2 5%) CAPAN-1: IMDM (GIBCO), FBS 20% (CO25%) The compounds tested in an in vivo model of oncology showed a significant level of activity.

PREPARATION EXAMPLE EXAMPLE A 2-Phenyl-2H-indazol-7-carboxamide (A6) Stage 1: methyl 3-methyl-2-nitrobenzoate (A1)

it was dissolved in EtOAc, washed several times with an ac solution. sat. NaHCO3 and brine and dried (Na2SO4). Evaporation of the solvent gave (A1) as a white solid that was used in the next step without further purification. 1H NMR (400 MHz, CDCl3, 300 K) δ 7.86 (1H, d, J = 7.5 Hz), 7.53-7.42 (2H, m), 3.89 (3H, s), 2.36 (3H, s). MS (ES) C9H9NO4 requires: 195, observed: 218 (M + Na) +.

Stage 2: 3- (Bromomethyl) -2-methyl nitrobenzoate (A2)

A mixture of (A1) (1.0 equiv.), (BzO) 2 (0.06 equiv.) And NBS (1.18 equiv.) In CCl4 (0.2 M with respect to A1) was heated to reflux in an atmosphere of N2 for 12 h. The mixture was cooled to RT, diluted with DCM, concentrated under reduced pressure while charging dry on SiO2. The residue was purified by flash column chromatography on SiO2 using 10:90 EtOAc / petroleum ether, yielding the desired (A2) as a white solid. 1H NMR (400 MHz, CDCl3, 300 K) δ 7.93 (1H, d, J = 7.7 Hz), 7.72 (1H, d, J = 7.7 Hz), 7.57 (1H, t, J = 7.7 Hz), 4.43 (2H, s), 3.88 (3H, s). MS (ES) C9H8BrNO4 requires: 273: 275, observed: 242: 244 (M-MeO) +, 227: 229 (M-NO2) +.

Stage 3: methyl 3-Formyl-2-nitrobenzoate (A3)

To a mixture of (A2) (1.0 equiv.) And 4 Å of sieves mol. (15 g) in MeCN (0.2 M) at RT NMMO (2.0 equiv.) Was added and the reaction mixture was stirred for 1.5 h under an N2 atmosphere. Then, the mixture was diluted with EtOAc, filtered and the filtrate was washed with H2O, 1N HCl and brine and dried (Na2SO4). Evaporation of the solvent gave (A3) as a white solid that was used in the next step without further purification. 1H NMR (400 MHz, CDCl3, 300 K) δ 9.96 (1H, s), 8.26 (1H, d, J = 7.9 Hz), 8.18 (1H, d, J = 7.9 Hz), 7.77 (1H, t, J = 7.9 Hz), 3.93 (3H, s). MS (ES) C9H7NO5 requires: 209, observed: 208 (M-H) -.

Stage 4: 2-Nitro-3 - [(phenylimino) methyl] methyl benzoate (A4)

A mixture of (A3) (1.0 equiv.) And aniline (1.05 equiv.) In EtOH (0.2 M) was stirred at reflux temperature under an atmosphere of N2 for 2 h until analysis by TLC revealed that the reaction was complete (Hexane / EtOAc = 75:25). Evaporation of the solvent gave (A4) as a white solid that was used in the next step without further purification. 1H NMR (400 MHz, CDCl3, 300 K) δ 8.51 (1H, d, J = 7.3 Hz), 8.41 (1H, s), 8.11 (1H, d, J = 7.8 Hz), 7.67 (1H, t, J = 7.8 Hz), 7.43 (2H, t, J = 7.8 Hz), 7.31 (1H, t, J = 7.3 Hz) , 7.16 (2H, d, J = 7.8 Hz), 3.94 (3H, s).

Stage 5: methyl 2-Phenyl-2H-indazol-7-carboxylate (A5)

A mixture of (A4) (1.0 equiv.) And NaN3 (1.05 equiv.) In dry DMF (0.3 M) was stirred at 90 ° C overnight in an N2 atmosphere. The crude product was reduced in vacuo and the residue was purified by flash column chromatography on silica using a gradient of EtOAc / petroleum ether from 10:90 to 40:60, yielding the desired (A5) as an oil of Dun. 1H NMR (400 MHz, CDCl3 300 K) δ 8.50 (1H, s), 8.12 (1H, d, J = 7.0 Hz), 7.96-7.90 (3H, m), 7 , 49 (2H, t, J = 7.6 Hz), 7.38 (1H, t, J = 7.4 Hz), 7.15 (1H, t, J = 7.4 Hz), 4.03 (3H, s). MS (ES) C15H12N2O2 requires: 252, observed: 253 (M + H) +.

Stage 6: 2-Phenyl-2H-indazol-7-carboxamide (A6)

The ester (A5) was heated in a mixture of THF and an ac solution. 32% NH3 at 70 ° C overnight in a tightly sealed tube. The solvents were reduced in vacuo and the residue was purified by flash column chromatography on silica using a gradient of EtOAc / petroleum ether from 30:70 to 50:50, yielding the desired (A6) as a white solid. . 1H NMR (400 MHz, DMSO, 300 K) δ 9.33 (1H, s), 8.56 (1H, sa), 8.16 (2H, d, J = 7.9 Hz), 8.08- 8.00 (2H, m), 7.88 (1H, sa), 7.63 (2H, t, J = 7.7 Hz), 7.50 (1H, t, 7.4 Hz), 7, 27 (1H, t, J = 7.9 Hz). MS (ES) C14H11N3O requires: 237, observed: 238 (M + H) +.

REPRESENTATIVE EXAMPLES

EXAMPLE 1

3- {4- [7- (aminocarbonyl) -2H-indazol-2-yl] phenyl} piperidinium chloride (B4)

Step 1: 3- [4 - ({- [3- (Methoxycarbonyl) -2-nitrophenyl] methylene} amino) phenyl] piperidine-1-carboxylic acid tert-butyl ester (B1)

(B1) was prepared following the general procedure indicated for Preparative Example A, step 4 using A3 and 3-4-aminophenyl) piperidine-1-carboxylate until TLC analysis revealed that the reaction had been completed ( Petroleum ether: EtOAc = 4: 1) and was used in the next step without further purification.

Stage 2: 2- {4- [1- (tert-Butoxycarbonyl) piperidin-3-yl] phenyl} -2H-indazol-7-methyl carboxylate (B2)

CDCl3, 300 K) δ 8.51 (1H, s), 8.13 (1H, d, J = 7.1 Hz), 7.95 (1H, d, J = 8.3 Hz), 7.91 (2H, d, J = 8.4 Hz), 7.39 (2H, d, J = 8.4 Hz), 7.18 (1H, t, J = 7.2 Hz), 4.30-4 , 10 (2H, m), 4.00 (3H, s), 2.85-2.70 (3H, m), 2.11-2.03 (1H, m), 1.83-1.75 (1H, m), 1.73-1.53 (2H, m overlapped with the H2O signal), 1.48 (9H, s). MS (ES) C25H29N3O4 requires: 435, observed: 436 (M + H) +.

Step 3: 3- {4- [7- (Aminocarbonyl) -2H-indazo] -2-yl) phenyl} piperidine-1-carboxylic acid tert-butyl ester (B3)

(B2) was heated in 7 N NH3 in MeOH (0.1 M) in a sealed tube for 2 days at 60 ° C. The solvents were reduced in vacuo and the crude product was purified by trituration with Et2O, giving the desired (B3) as a yellow solid. 1H NMR (400 MHz, CDCl3, 300 K) δ 9.04 (1H, sa), 8.51 (1H, s), 8.31 (1H, d, J = 6.8 Hz), 7.91 ( 1H, d, J = 8.3 Hz), 7.84 (2H, d, J = 8.2 Hz), 7.42 (2H, d, J = 8.2 Hz), 7.31-7, 22 (1H, m overlapped to the CDCl3 signal), 5.95 (1H, sa), 4.40-4.05 (2H, m), 2.90-2.70 (3H, m), 2, 15-2.00 (1H, m), 1.85-1.75 (1H, m), 1,751.50 (2H, m overlapped with the H2O signal), 1.48 (9H, s). MS (ES) C24H28N4O3 requires: 420, observed: 421 (M + H) +.

Stage 4: 3- {4- [7- (aminocarbonyl) -2H-indazol-2-yl] phenyl} piperidinium chloride (B4)

To a stirred solution of (B3) (1.0 equiv.) In EtOAc (0.2 M) was added a solution of 4 N HCl / 1,4-dioxane (10.0 equiv.) And the reaction mixture stirred at RT for 3 h. The solvent was evaporated under reduced pressure and the crude product was purified by trituration with Et2O, yielding the desired (B4) as a yellow solid. 1H NMR (400 MHz, DMSO-d6 300 K) δ 9.32 (1H, s), 9.12 (1H, sa), 8.87 (1H, sa), 8.55 (1H, sa), 8 , 13 (2H, d, J = 8.6 Hz), 8.06 (1H, J = 7.0 Hz), 8.02 (1H, d, J = 8.4 Hz), 7.89 (1H , sa), 7.55 (2H, d, J = 8.6 Hz), 7.27 (1H, dd, J = 8.4, 7.0 Hz), 3.43-3.27 (2H, m), 3.17-3.03 (2H, m), 3.00-2.85 (1H, m), 2.00-1.70 (4H, m). MS (ES) C19H21ClN4O requires: 320, observed: 321 (M + H) +.

EXAMPLE 2

2- {4 - [(3R) -Piperidin-3-yl] phenyl} -2H-indazol-7-carboxamide (C1) and 2- {4 - [(3S) -piperidin-3-yl] phenyl} -2H -indazol-7carboxamide (C2)

Example 1, B4 was separated by chiral SFC (column: Chiralpak AS-H, 1 x 25 mm, flow: 10 ml / min, Tcol: 35C, Pcol: 100 bar, modifier: 55% (1PrOH + 4% Et2NH )), using CO2 as a supercritical eluent, producing both pure enantiomers.

The first eluted enantiomer (C1), retention time (SFC): 4.80 min, was obtained as a white powder. 1H NMR (400 MHz, DMSO-d6, 300 K) δ 9.28 (s, 1H), 8.57 (sa, 1H), 8.06 (d, 2H, J = 7.2 Hz), 8, 04 (d, 2H, J = 8.4 Hz), 7.88 (sa, 1H), 7.49 (d, 2H, J = 8.4 Hz), 7.27 (dd, 1H, J = 8 , 4, 7.2 Hz), 3.08-2.94 (m, 2H), 2.77-2.67 (m, 1H), 2.64-2.52 (m, 1H), 1, 98-1.90 (m, 1H), 1.75-1.47 (m, 4H). MS (ES) G19H20N4O requires: 320, observed: 321 (M + H) +. The free base was converted to (3R) -3- {4- [7- (aminocarbonyl) -2H-indazol-2-yl] phenyl} piperidinium chloride and the optical rotation was measured: [α] 20D = +133, 3 (c 0.15, MeOH).

The second eluted enantiomer (C2), retention time (SFC): 6.51 min, was obtained as a white powder. 1H NMR (400 MHz, DMSO-d6, 300 K) δ 9.28 (s, 1H), 8.57 (sa, 1H), 8.06 (d, 2H, J = 7.2 Hz), 8, 04 (d, 2H, J = 8.4 Hz), 7.88 (sa, 1H), 7.49 (d, 2H, J = 8.4 Hz), 7.27 (dd, 1H, J = 8 , 4, 7.2 Hz), 3.08-2.94 (m, 2H), 2.77-2.67 (m, 1H), 2.64-2.52 (m, 1H), 1, 98-1.90 (m, 1H), 1.75-1.47 (m, 4H). MS (ES) C19H20N4O requires: 320, observed: 321 (M + H) +. The free base was converted to (3S) -3- {4- [7- (aminocarbonyl) -2H-indazol-2-yl] phenyl) piperidinium chloride and the optical rotation was measured: [α] 20D = -137, 9 (c 0.145, MeOH).

EXAMPLE 3

3- {4- [7- (Aminocarbonyl) -5-fluoro-2H-indazol-2-yl] phenyl} piperidinium trifluoroacetate (D4)

Stage 1: methyl 5-Fluoro-1H-indazol-7-carboxylate (D1)

To a solution of Example 4, E3 (1.0 equiv.) In 1,2-dichloroethane (0.1 M) was added AcCl (5 equiv.) And heated at 55 ° C for 2 h. Then, the solvent was removed under reduced pressure.

The white solid was dissolved in toluene / water (5/1, 0.1 M). The solution was cooled to 0 ° C and HCl (10 equiv., 37%) was added. Then, NaNO2 (10 equiv.) Was added slowly and portionwise and the mixture was stirred for 3 h at 0 ° C. The organic phase was washed with water (3 x), dried over MgSO4 and the solvent was removed under reduced pressure.

Then, the yellow solution in toluene (0.1 M) was heated for 2 h at 90 ° C. Evaporation of toluene provided the desired product as a red solid. 1H NMR (400 MHz, DMSO, 300 K) δ 13.37 (1H, s), 8.23 (1H, s), 7.63 (1H, dd, J = 8.6 Hz, J = 2.5 Hz), 7.48 (1H, dd, J = 8.6 Hz, J = 2.5 Hz), 3.66 (3H, s). MS (ES +) C9H7FN2O2 requires: 194, observed: 195 (M + H) +.

Stage 2: 5-Fluoro-1H-indazol-7-carboxamide (D2)

(D1) was dissolved in dioxane / water (1/1, 0.1 M) and KOH (1.5 equiv.) Was added. After stirring 12 h at RT, the solvents were removed under reduced pressure. The white solid was used without purification for subsequent coupling.

The carboxylic acid was dissolved in DMF (0.1 M) and TBTU (1.5 equiv.) Was added at 0 ° C. After 15 min, DIPEA (2.0 equiv.) And ammonia (3.0 equiv., 0.5 M in dioxane) were added and the mixture was stirred 36 h at RT. EtOAc was added and the organic phase was washed with an aq solution. sat. NaHCO3 (3 x) and brine (2 x). The organic phase was dried and evaporated under reduced pressure. The crude product was purified by flash chromatography using 1-20% MeOH / DCM, yielding (D2) as a white solid. MS (ES +) C8H6FN3O requires: 179, observed: 180 (M + H) +.

Stage 3: 2- (4-Bromophenyl) -5-fluoro-2H-indazol-7-carboxamide (D3)

To a solution of D2 (1.0 equiv.) In DMF (0.2 M) K2CO3 (1.3 equiv.) And 4-bromofluorobenzene (10.0 equiv.) Were added and the reaction mixture was heated in microwave conditions at 180 ° C for 20 min. The reaction mixture was cooled to RT and diluted with EtOAc. The organic phase was washed with brine; dried (Na2SO4). Evaporation of the solvent gave (D3) which was purified by chromatography on silica gel eluting with 5070% EtOAc / petroleum ether, obtaining the title compound as a yellow powder. 1H NMR (400 MHz, DMSO-d6, 300 K) δ 9.34 (1H, s), 8.50 (1H, sa), 8.17 (2H, d, J = 9.0 Hz), 8, 03 (1H, sa), 7.90-7.80 (4H, m). MS (ES +) C14H9BrFN3O requires: 334/336, observed: 335/337 (M + H) +.

Stage 4: 5-Fluoro-2- (4-pyridin-3-ylphenyl) -2H-indazol-7-carboxamide (D4)

A mixture of (D3) (1.0 equiv.) And pyridine-3-boronic acid (1.3 equiv.) In DMF (1.0 M) together with a 2 N solution

-

Na2CO3 (2.0 equiv.) was degassed with an Ar current for 30 min. TBu3PH + BF4 (0.05 equiv.) And Pd2 (dba) 3 (0.05 equiv.) Were added and the reaction mixture was heated at 90 ° C for 48 h. The mixture was cooled to RT, DCM was added and the organic phase was washed with an aq solution. sat. NaHCO3 and brine and dried (Na2SO4). The solution was concentrated under reduced pressure and the residue was purified by chromatography on silica gel eluting with 50-90% EtOAc / petroleum ether and then 10% MeOH / DCM, obtaining the title compound as a powder of yellow color. 1H NMR (400 MHz, DMSO-d6, 300 K) δ 9.40 (1H, s), 9.01 (1H, d, J = 1.6 Hz), 8.63 (1H, dd, J = 4 , 8, 1.6 Hz), 8.57 (1H, sa), 8.32 (2H, d, J = 8.8 Hz), 8.20 (1H, d, J = 7.8 Hz), 8.10 (1H, sa), 8.01 (2H, d, J = 8.8 Hz), 7.88-7.82 (2H, m), 7.54 (1H, dd, J = 7, 8, 4.8 Hz). MS (ES) C19H13FN4O requires: 332, observed: 333 (M + H +).

Step 5: 3- {4- [7- (Aminocarbonyl) -5-fluoro-2H-indazol-2-yl] phenyl} piperidine-1-carboxylate benzyl (D5)

To a stirred solution of (D4) in dry MeOH (0.2 M) was added NaBH4 (1.2 equiv.) And then dropwise Cbz-Cl (1.2 equiv.) At -65 ° C. The reaction was allowed to reach RT overnight and then was interrupted with H2O. MeOH was concentrated under reduced pressure and EtOAc was added. The organic phase was washed with an ac solution. sat. NaHCO3 and dried (Na2SO4). Evaporation of the solvent gave (D5) which was used in the next step without further purification. MS (ES) C27H25FN4O3 requires: 472, observed: 473 (M + H +).

Step 6: 3- {4- [7- (aminocarbonyl) -5-fluoro-2H-indazol-2-yl] phenyl} piperidinium trifluoroacetate (D6)

To a solution of (D5) (1.0 equiv.) In MeOH (0.2 M) was added 10% Pd / C (0.05 equiv.) And HCl (1.0 equiv.) And the mixture The reaction was stirred under an atmosphere of H2 (1 atm) for 48 h. Then, the mixture was filtered through Celite and the solvent was removed in vacuo to provide (D6) which was purified by reverse phase RP-HPLC (column: C18), using H2O (0.1% TFA) and MeCN. (0.1% TFA) as eluents, the desired fractions were lyophilized to provide the title compound (D6) as a white powder. 1H NMR (400 MHz, CD3CN, 300 K) δ 9.28 (1H, s), 8.89 (1H, sa), 8.60-8.50 (2H, m), 8.13 (2H, d , J = 8.6 Hz), 8.09 (1H, sa), 7.90-7.70 (2H, m), 7.54 (2H, d, J = 8.6 Hz), 3.40 -3.30 (2H, m), 3.20-2.80 (3H, m), 2.00.1.90 (2H, m), 1.80-1.70 (2H, m), MS (EN) C19H19FN4O requires: 338, observed: 339 (M + H +).

EXAMPLE 4

5-Fluoro-2- (3-fluoro-4-piperidin-3-ylphenyl) -2H-indazol-7-carboxamide trifluoroacetate (E6)

Stage 1: 5-Fluoro-3-methyl-2-nitrobenzoic acid (E1)

To a solution of 3-fluoro-5-methylbenzoic acid (1.0 equiv.) In conc. H2O4. KNO3 (1.1 equiv.) was added slowly at 0 ° C. The mixture was stirred at RT for 1 h and then slowly poured into ice-cold water. After stirring until the ice melted completely, the white precipitate was filtered, washed with cold water and dried under reduced pressure. The white solid was used without further purification for the next step. 1H NMR (400 MHz, DMSO, 300 K) δ 14.08 (1H, s a), 7.65 (2H, m), 2.30 (3H, s).

Stage 2: methyl 5-Fluoro-3-methyl-2-nitrobenzoate (E2)

To a solution of (E1) and cesium carbonate (1.5 equiv.) In DMF (0.25 M) at RT was added methyl iodide (1.0 equiv.). After the mixture was stirred for 18 h, brine was added and the mixture was extracted with EtOAc. The organic phase was dried (Na2SO4) and concentrated under reduced pressure. The yellow solid was used in the next step without purification. 1H NMR (400 MHz, DMSO, 300 K) δ 7.63 (2H, m), 3.83 (3H, s), 2.29 (3H, s).

Stage 3: methyl 2-Amino-5-fluoro-3-methylbenzoate (E3)

A mixture of (E2) (1.0 equiv.) And Pd / C (10% w / w) in MeOH (0.25 M) was stirred for 3 d at RT under an atmosphere of H2 (1 atm). The mixture was filtered through Celite® and then the solvent was evaporated under reduced pressure. The white solid was used without further purification in the subsequent step. 1H NMR (400 MHz, DMSO, 300 K) δ 7.29 (1H, dd, J = 9.5 Hz, J = 3.0 Hz), 7.12 (1H, dd, J = 9.5 Hz, J = 3.0 Hz), 6.36 (2H, sa), 3.78 (3H, s), 2.11 (3H, s).

Stage 4: methyl 2,5-Difluoro-3-methylbenzoate (E4)

To a solution of (E3) (1.0 equiv.) In dry DCM (0.4 M) at 0 ° C was added portionwise nitrosonium tetrafluoroborate (1.3 equiv.). After 1 h at 0 ° C, dry dichlorobenzene (120 equiv.) Was added and the reaction was slowly heated to 160 ° C while DCM distillation was removed. After 3 h, the mixture was cooled to RT, EtOAc was added and the organic phase was washed with brine (2 x). After drying over MgSO4, the solvents were removed under reduced pressure. The crude product was purified by flash chromatography using 1-10% EtOAc / petroleum ether, yielding (E4) as a yellow oil. 1H NMR (400 MHz, CDCl3, 300 K) δ 7.42 (1H, m), 7.06 (1H, m), 3.92 (3H, s), 2.30 (3H, d, J = 2 , 3 Hz).

Stage 5: methyl 2,5-Difluoro-3-formylbenzoate (E5)

(E5) was prepared from E4 following the general procedure indicated in Preparative Example A steps 2 and 3. The crude product was purified by flash chromatography EtOAc 1-20% / petroleum ether, yielding a white solid. . 1H NMR (300 MHz, DMSO, 300 K) δ 10.19 (1H, d, J = 2.4 Hz), 7.98 (1H, m), 7.86 (1H, m), 3.89 ( 3H, s). MS (ES +) C9H6F2O3 requires: 200, observed: 201 (M + H) +.

Step 6: 5-Fluoro-2- (3-fluoro-4-piperidin-3-ylphenyl) -2H-indazol-7-carboxamide trifluoroacetate (E6)

(E5) was converted into the corresponding indazole using tert-butyl 3- (4-amino-2-fluorophenyl) piperidine-1-carboxylate following the general procedure indicated in Preparative Example A steps 4 and 5.

The resulting methyl 2- {4- [1- (tert-butoxycarbonyl) piperidin-3-yl] -3-fluorophenyl} -5-fluoro-2H-indazol-7-carboxylate was further converted into the corresponding carboxamide by treatment with KOH (1.3 equiv.) In dioxane / water (0.1 M) for 12 h at RT. The solvents were removed under reduced pressure. The carboxylic acid was dissolved in DMF (0.1 M) and TBTU (1.5 equiv.) Was dissolved. After 15 min, DIPEA (2.0 equiv.) And ammonia (3.0 equiv., 0.5 M in THF) were added and the solution was stirred for 36 h. The mixture was diluted with EtOAc and then the organic phase was washed with an aq solution. sat. NaHCO3 and brine. After evaporation of the solvent the residue was used in the next step without purification.

For deprotection, the crude product was dissolved in TFA / DCM (0.1 M) and stirred for 3 h at RT. Evaporation of the solvent gave a residue that was purified by reverse phase HPLC (column: C18) to give the title compound (E6). 1H NMR (400 MHz, DMSO, 300 K) δ 9.34 (1H, s), 8.90 (1H, m), 8.61 (1H, m), 8.49 (1H, s), 8, 18 (1H, dd, J = 11.6 Hz, 2.0 Hz), 8.05 (2H, m), 7.81 (2H, m), 7.63 (1H, m), 3.34 ( 3H, m), 3.13 (1H, m), 2.94 (1H, m), 1.95-1.76 (4H, m). MS (ES +) C19H18F2N4O requires: 356, observed: 357 (M + H) +.

EXAMPLE 5

(3S) -3- {4- [7- (Aminocarbonyl) -2H-indazol-2-yl] phenyl} piperidinium 4-methylbenzenesulfonate (F4)

Stage 1: (3S) -3- [4 - ({(IE) - [3- (methoxycarbonyl) -2-nitrophenyl] methylene} amino) phenyl] piperidine-1-carboxylic acid tert-butyl ester (F1)

(F1) was prepared from A3 and (3S) -3- (4-aminophenyl) piperidine-1-carboxylic acid tert-butyl ester (prepared by the resolution of 3- (4-aminophenyl) -piperidine with 2 equivalents of acid L-dibenzoyl tartaric in MeOH and subsequent Boc-protection) as described in Example 1, B1.

Stage 2: 2- {4 - [(3S) -1- (tert-butoxycarbonyl) piperidin-3-yl] phenyl} -2H-indazol-7-carboxylic acid (F2)

(F1) (1 equiv.) And sodium azide (1 equiv.) Were suspended in DMF (0.25 M), in an inert state and 2,6-lutidine (1.0 equiv.) Was added. The mixture was heated at an internal temperature of 110 ° C for 20 hours. The resulting brown solution was cooled to 20 ° C and THF and a 25% by weight aqueous solution of LiCl was added. The phases were separated and the organic product was washed three more times with a 25% by weight aqueous solution of LiCl. To the above organic solution was added 2.0 M NaOH (10 equiv.), The mixture was heated at 35 ° C for 20 hours before cooling to 20 ° C and the phases were separated. The organic phase was washed with a mixture of 2.0M HCl acid and brine, the phases were separated, the organic phase was further washed with brine and concentrated, giving (F2) that was not further purified.

Step 3: (3S) -3- {4- [7- (Aminocarbonyl) -2H-indazol-2-yl] phenyl} piperidine-1-carboxylic acid tert-butyl ester (F3)

F2 was dissolved in DCM (0.35 M) and di tert-butyl carbonate (1.3 equiv.) And pyridine (1.0 equiv.) Were added at RT.

5 After 30 minutes, ammonium bicarbonate (1.3 equiv.) Was added and stirring continued for 20 hours. 1M HCl (5 ml / g) was added, the phases were separated, the organic phase was washed twice with water and concentrated, giving a low volume. The crude compound (F3) was filtered through a layer of silica and then crystallized from methyl tert-butyl ether.

Step 4: (3S) -3- {4- [7- (aminocarbonyl) -2H-indazol-2-yl] phenyl} piperidinium (F4) 4-Methylbenzenesulfonate (F4)

10 F3 was dissolved in THF (0.15 M) and water (5% compared to THF) was added. Para-toluene sulfonic acid monohydrate (2.2 equiv.) Was added and the mixture was heated to 66 ° C and stirred overnight. After cooling the desired solid salt was isolated by filtration and confirmed to be a monohydrate (F4). 1H NMR (400 MHz, DMSO, 300 K) δ 9.34 (1H, s); 9.20 (1H, wide s), 8.58 (1H, s), 8.14 (2H, d, J = 8.8 Hz), 8.05 (2H, ddd, J = 1.2, 7 , 2, 16.8 Hz), 7.93 (1H, s), 7.52 (4H, dd, J = 8.8, 16.8 Hz), 7.27 (1H, dd, J = 6, 8, 8.0 Hz), 7.13 (2H, d, J = 8 Hz), 3.48 (3H, m), 3.10

15 (2H, m), 2.90 (1H, m); 2.30 (3H, s), 1.89 (2H, m), 1.75 (2H, m).

The following examples were prepared according to the procedures of the previous examples:

Example

Name MW M + H + Example Procedure

6

3- {4- [7- (Aminocarbonyl) -2H-indazol-2yl] phenyl} piperidinium trifluoroacetate 320 321 one

7

5-Fluoro-2- (4-piperidin-3-ylphenyl) -2H-indazol-7-carboxamide 338 339 3

8

(3S) -3- {4- [7- (aminocarbonyl) -2H-indazol-2yl] phenyl} piperidinium chloride 320 321 2

9

(3R) -3- {4- [7- (aminocarbonyl) -2H-indazol-2yl] phenyl} piperidinium chloride 320 321 2

10

(R) -5-Fluoro-2- (4-piperidin-3-ylphenyl) -2H-indazol-7carboxamide 338 339 2

eleven

(S) -5-Fluoro-2- (4-piperidin-3-ylphenyl) -2H-indazol-7-carboxamide 338 339 2

12

(R) -5-Fluoro-2- {3-fluoro-4-piperidin-3-ylphenyl} -2H-indazol-7carboxamide 356 357 2

13

(S) -5-Fluoro-2- {3-fluoro-4-piperidin-3-ylphenyl} -2H-indazol-7carboxamide 356 357 2

Claims (20)

1. A compound of formula I: image 1 in which: R1 is hydrogen or fluorine; Y R2 is hydrogen or fluorine; or pharmaceutically acceptable salts, stereoisomers or tautomers thereof. 2. A compound of claim 1 of formula II: image 1 Wherein R1 and R2 are as defined in claim 1; or pharmaceutically acceptable salts, stereoisomers or tautomers thereof. 3. A compound of claim 1 of formula III: image 1 wherein R1 and R2 are as defined in claim 1; 15 or pharmaceutically acceptable salts or tautomers thereof. 4. A compound of claim 1 of formula IV: image 1 5 10 fifteen twenty 25 30 wherein R1 and R2 are as defined in claim 1; or pharmaceutically acceptable salts or tautomers thereof.

5.

A compound of any previous claim wherein R1 is hydrogen and R2 is hydrogen or fluorine.

6.

A compound of claim 1 selected from: 2- (4-piperidin-3-ylphenyl) -2H-indazol-7-carboxamide; 2- {4 - [(3R) -piperidin-3-yl] phenyl} -2H-indazol-7-carboxamide; 2- {4 - [(3S) -piperidin-3-yl] phenyl} -2H-indazol-7-carboxamide; 5-fluoro-2- (4-piperidin-3-ylphenyl) -2H-indazol-7-carboxamide; 5-fluoro-2- {4 - [(3S) -piperidin-3-yl] phenyl} -2H-indazol-7-carboxamide; 5-fluoro-2- {4 - [(3R) -piperidin-3-yl] phenyl} -2H-indazol-7-carboxamide; 5-fluoro-2- (3-fluoro-4-piperidin-3-ylphenyl) -2H-indazol-7-carboxamide; 5-fluoro-2- {3-fluoro-4 - [(3R) -piperidin-3-yl] phenyl} -2H-indazol-7-carboxamide; 5-fluoro-2- {3-fluoro-4 - [(3S) -piperidin-3-yl] phenyl} -2H-indazol-7-carboxamide;

and pharmaceutically acceptable salts, tautomers, stereoisomers thereof.

7.

A compound of claim 6 which is: 2- (4-piperidin-3-ylphenyl) -2H-indazol-7-carboxamide;

or a pharmaceutically acceptable salt or tautomer thereof. 8. A compound of claim 6 which is: 2- {4 - [(3R) -piperidin-3-yl] phenyl} -2H-indazol-7-carboxamide; or a pharmaceutically acceptable salt or tautomer thereof. 9. A compound of claim 6 which is: 2- {4 - [(3S) -piperidin-3-yl] phenyl} -2H-indazol-7-carboxamide; or a pharmaceutically acceptable salt or tautomer thereof.

10.

A pharmaceutical composition comprising a compound of any prior claim or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof in association with a pharmaceutically acceptable carrier.

eleven.

A compound of any one of claims 1 to 9, or a pharmaceutically acceptable salt, stereoisomer or tautomer thereof and an antineoplastic agent for simultaneous, separate or sequential administration.

12.

A compound of any one of claims 1 to 9, or a pharmaceutically acceptable salt, stereoisomer or tautomer thereof, for use in therapy.

13.

The use of a compound of any one of claims 1 to 9 or a pharmaceutically acceptable salt, stereoisomer or tautomer thereof for the manufacture of a medicament for the treatment or prevention of

conditions that can be relieved by the inhibition of poly (ADP-ribose) polymerase (PARP).

14.

The use of a compound of any one of claims 1 to 9 or a pharmaceutically acceptable salt, stereoisomer or tautomer thereof for the manufacture of a medicament for the treatment or prevention of cancer, inflammatory diseases, reperfusion lesions, ischemic conditions, stroke, renal failure, cardiovascular diseases, vascular diseases other than cardiovascular diseases, diabetes, neurodegenerative diseases, retroviral infection, retinal damage or senescence of the skin and UV-induced skin damage.

fifteen.

The use of a compound of any one of claims 1 to 9 or a pharmaceutically acceptable salt, stereoisomer or tautomer thereof for the manufacture of a medicament for the treatment or prevention of cancer.

16.

The use of claim 15 wherein the cancer is a cancer that is deficient in DSB repair activity of Homologous Recombination (RH) dependent DNA.

17.

A compound of any one of claims 1 to 9 or a pharmaceutically acceptable salt, stereoisomer or tautomer thereof for use in the treatment or prevention of conditions that can be alleviated by the inhibition of poly (ADP-ribose) polymerase (PARP).

18.

A compound of any one of claims 1 to 9 or a pharmaceutically acceptable salt, stereoisomer or tautomer thereof for use in the treatment or prevention of cancer, inflammatory diseases, reperfusion lesions, ischemic conditions, stroke, renal failure, cardiovascular diseases , vascular diseases other than cardiovascular diseases, diabetes, neurodegenerative diseases, retroviral infection, retinal damage or senescence of the skin and UV-induced skin damage.

19.

A compound of any one of claims 1 to 9 or a pharmaceutically acceptable salt, stereoisomer or tautomer thereof for use in the treatment or prevention of cancer.

twenty.

A compound of any one of claims 1 to 9 or a pharmaceutically acceptable salt, stereoisomer or tautomer thereof for use in the treatment or prevention of cancer that is deficient in DSB repair activity of Homologous Recombination (RH) dependent DNA.