MXPA01002603A - 2-oxo-imidazolidine-4-carboxylic acid hydroxamine compounds that inhibit matrix metalloproteinases - Google Patents

Abstract

The present invention relates to a compound of the formula wherein R<1>, R<2>, R<3>and R<4>are as defined above, and pharmaceutically acceptable salts and solvates thereof, that are useful, for example, as matrix metalloproteinase inhibitors. The present invention is also directed to pharmaceutical compositions comprising such compounds and methods of treatment for diseases such as osteoarthritis, rheumatoid arthritis, cancer, osteoporosis, tissue ulceration, restinosis, periodontal disease, inflammation, epidermolysis bullosa, scleritis, stroke, Alzheimer's disease, and the like, characterized by inappropriate matrix metalloproteinase activity. Processes for the synthesis of compounds of formula (I) are also disclosed.

Description

HYDROXAMIDE COMPOUNDS OF 2-OXO-IMIDAZOLIDINE-4-CARBOXYLIC ACIDS THAT INHIBIT METALOPROTEINASE FROM THE MATRIX BACKGROUND OF THE INVENTION The present invention relates to hydroxamide derivatives of 2-oxo-imidazolidine-4-carboxylic acids, to pharmaceutical compositions comprising such derivatives, and to the use of such derivatives in the treatment of arthritis, inflammation, cancer and other diseases which are described later. The compounds of the present invention are inhibitors of zinc metalloendopeptidases, especially those belonging to the subfamilies of the meticincins constituted by the matrix metalloproteinases (also called MMP or matp'xinas) and the reprolysins (also known as adamilsinas) ( Rawlings et al., Methods in Enzvmology, 248, 183-228 (1995) and Stocker et al., Protein Science, 4, 823-840 (1995)). The subfamily of MMP enzymes, currently contains seventeen members (MMP-1, MMP-2, MMP-3, MMP-7, MMP-8, MMP-9, MMP-10, MMP-11, MMP-12, MMP-13) , MMP-14, MMP-15, MMP-16, MMP-17, MMP-18, MMP-19 and MMP-20). MMPs are best known for their role in regulating the turnover of extracellular matrix proteins and therefore play important roles in normal physiological processes such as reproduction, development and differentiation. In addition, MMPs are expressed in many disease states where abnormal connective tissue turnover occurs. For example, it has been shown that MMP-13, an enzyme with potent activity in the degradation of type II collagen (the main collagen of cartilage), is overexpressed in osteoarthritic cartilage (Mitchell et al., J. Clin. ., 97,761 (1996)). Other MMPs (MMP-2, MMP-3, MMP-8, MMP-9, MMP-12) are also overexpressed in osteoarthritic cartilage and it is expected that the inhibition of some or all of these MMPs will retard or block the accelerated loss of cartilage, typical of diseases of the joints such as osteoarthritis or rheumatoid arthritis. Overexpression of certain metalloproteinases is also associated with the metastasis of tumor cells. It is thought that such activity is essential for the invasion of healthy tissues. It is expected that the inhibition of the activity of some or all of these proteinases limits the spread of malignant cells. Additionally, certain metalloproteinases are necessary for angiogenesis, the process by which a growing tumor, for example, obtains an additional supply of blood by means of a new vascularization. Therefore, it is expected that the inhibition of these enzymes retards or stops tumor growth. It is also to be expected that the compounds of the invention usefully inhibit additional classes of enzymes that play important roles in both normal and pathological processes. For example, mammalian reprolysins are known as ADAMs (A Disintegrin And Metalloproteinase) (Wolfberg et al., Cell. Biol. 131, 275-278 (1995)) and contain a disintegrin domain in addition to a disintegrin and metalloproteinase. of a domain similar to that of metalloproteinases. To date, twenty-three different ADAMs have been identified. ADAM-17, also known as tumor necrosis factor alpha (TACE) conversion enzyme, is the best known ADAM. ADAM-17 (TACE) is responsible for the cleavage of the alpha factor from tumor necrosis attached to cells (TNF-a, also known as cachectin). It has been recognized that TNF-a is implicated in many infectious and autoimmune diseases (W. Friers, FEBS Letters, 285, 199 (1991)). In addition, it has been shown that TNF-a is the main mediator of the inflammatory response observed in sepsis and septic shock. (Spooner et al., Clinical Immunoloqy and Immunopatholoqv, 62 S11 (1992)). There are two forms of TNF-α, a membrane protein of type II of relative molecular mass 26,000 (26 kD) and a soluble form of 17 kD generated from the protein bound to cells by specific proteolytic cleavage. The soluble form of 17 kD and TNF-a is released by the cell and is associated with the deleterious effects of TNF-a. This form of TNF-a is also capable of acting at sites distant from the synthesis site. So that, inhibitors of TACE prevent the formation of soluble TNF-a and prevent the harmful effects of soluble factor. In another example, aggrecanase, in an enzyme that is important in the degradation of cartilage aggrecan. It is believed that aggrecanase is an ADAM. The loss of aggrecan from the cartilage matrix is an important factor in the progression of joint diseases such as osteoarthritis and rheumatoid arthritis, and it is expected that the inhibition of aggrecanase will retard or block the loss of cartilage in tissues affected by these. diseases. Other ADAMs that have shown expression in disease states include ADAM TS-1 (Kuno et al., J. Biol. Chem., 272, 556-562 (1997)), and ADAMs 10, 12 and 15 (Wu et al. ., Biochem, Biophys, Res. Comm .. 235, 437-442 (1997)). As knowledge of the expression, it will be appreciated that the association of physiological substrates and diseases, of the ADAMs, increases the total significance of the role of the inhibition of this class of enzymes. The compounds of the invention are useful in the treatment of arthritis (including osteoarthritis and rheumatoid arthritis), inflammatory bowel disease, Crohn's disease, emphysemas, acute respiratory distress syndrome, asthma, chronic obstructive pulmonary disease, the disease Alzheimer's disease, organ transplant toxicity, cachexia, allergic reactions, inflammation, allergic contact hypersensitivity, cancer (such as cancers of solid tumors including colon cancer, breast cancer, lung cancer and cancer) prostate, as well as haematopoietic malignancies including leukaemias and lymphomas), tissue ulceration, restenosis, periodontal disease, epidermolysis bullosa, osteoporosis, loosening of artificial joint implants, atherosclerosis (including rupture of atherosclerotic plaques), aortic aneurysms ( including abdominal aortic aneurysm and cerebral aortic aneurysm), congestive heart failure, myocardial infarction, stroke, cerebral ischemia, head trauma, spinal cord damage, neuro-vegetative disorders (acute and chronic), autoimmune diseases, Huntington's disease, Parkinson's disease, migraine, depression, peripheral neuropathy, pain, cerebral amyloid angiopathy, nootropic or understanding increase, amyotrophic lateral sclerosis, multiple sclerosis, ocular angiogenesis, corneal damage, macular degeneration, abnormal wound healing, burns, diabetes, invasion tumor, tumor growth, tumor metastasis, corneal scarring, scleritis, AIDS, and sepsis or septic shock. The compounds of the present invention are also useful in the treatment of diseases in which the inhibition of MMPs and / or ADAMs can provide therapeutic benefit, such as the characteristics by expression of matrix metalloproteinases or expression of ADAM. They are well known, according to the scientific literature, inhibitors of matrix metalloproteinases and reprolysins. Specifically, European patent publication 606,046, published on July 13, 1994, alludes certain heterocyclic MMPs inhibitors. The publication of PCT documents WO 98/08825 and WO 98/08815, both published on March 5, 1998, refer to certain inhibitors of MMPs of the cyclic hydroxamic acid type. U.S. Patent No. 5,861, 510, issued January 19, 1999, alludes to cyclic arylsulfonylamino hydroxamic acids which are useful as inhibitors of MMPs. The PCT publication of WO 98/349148, published on August 13, 1988, alludes to cyclic hydroxamic acids including certain dialkyl substituted compounds that are useful as inhibitors of MMPs. The PCR publications of WO 96/27583 and WO 98/07697, published on March 1, 1996 and February 26, 1998, respectively, refer to alkylsulfonyl hydroxamic acids. The PCT publication of WO 98/03516, published on January 29, 1998, refers to phosphinates with MMP activity. PCT Publication 98/33768, published on August 6, 1998, refers to unsubstituted arylsulfonylaminohydroxamic acids in N. The publication of European Patent EP 935,963, published on August 18, 1999, refers to the use of selective inhibitors of MMP-13 for the treatment of osteoarthritis. The patent applications of E.U.A. 09 / 290,022, 09 / 287,930 and 09 / 287,508, filed on April 9, 1999, April 7, 1999 and April 7, 1999, respectively, refer to methods of preparation of hydroxamic acids. The provisional patent application of E.U.A. entitled "Selective Inhibitors of Aggrecanase in Osteoarthritis Treatment", presented on August 12, 1999, refers to inhibitors of MMPs, Agrecanase and TACE, and to additional methods of preparation of hydroxamic acids. The non-provisional patent application of E.U.A. entitled "TACE inhibitors", filed on August 12, 1999, refers to heterocyclic hydroxamic acids. Each of the aforementioned publications and patent applications is incorporated herein by reference in its entirety. It has been recognized that different combinations of MMPs and ADAMs are expressed in different pathological states. Therefore, inhibitors with specific selectivities for ADAMs and / or individual MMPs can be preferred for individual diseases. For example, rheumatoid arthritis is an inflammatory disease of the joints characterized by excessive levels of TNF, and loss of constituents of the joint matrix. In this case, a compound that inhibits TACE and aggrecanase as well as MMPs such as MMP-13, may be the preferred therapy. In contrast, in a less inflammatory joint disease, such as osteoarthritis, compounds that inhibit MMPs that degrade the matrix such as MMP-13, but not TACE, may be preferred.

BRIEF DESCRIPTION OF THE INVENTION The present invention relates to a compound according to formula (I) or a pharmaceutically acceptable salt or solvate thereof, wherein R1 is selected from the groups consisting of C6-C10 aryl, C1-C9 heteroaryl, aryl (C6-C? 0) CrC6 alkyl, heteroaryl (C? C9) C6 alkyl, C6-C10 aryl, C6-C10 aryl, C6-C6 heteroaryl, C6-C6 aryl, C6-C6 heteroaryl, heteroaryl (C? -C9) C1-C9 heteroaryl, aryl (C? -C?) Oxyaryl Ce-Cio, heteroaryl (C? -9) oxyaryl C6-C? 0, aryl (C6-C? O) oxyheteroaryl of C1-C9, (C1-C9) heteroaryl-C1-C9-oxyheteroaryl, C6-C6 (C6-C? o) oxalkyl, C6-C6 heteroaryl (C? -Cg) oxyalkyl, aryl (C6-C6) C6-C? O) alkyl (C? -C6) aryl of C6-C? 0, heteroaryl (C? -9) alkyl (C? -C6) aryl of C6-C? 0, aryl (C6-C ?o) C 1 -C 6 alkyl) C 1 -C 9 heteroaryl, heteroaryl (C?-C9) alkyl (C 1 -C) heteroaryl (C?-C 9), aryl (C 6 -C C) alkoxy ( C? -C6) C6-C? 0 aryl, heteroaryl (Cr C9) (C? -C6) alkoxy C6-C? 0 aryl, aryl (C6-C1o) (C? -C6) alkoxy heteroaryl C1-C9, heteroaryl (C-C9) alkoxy (C6-6) heteroaryl of C1-C9, aryl (C6-C6) oxyalkyl (C-C6) aryl of C6-C6O, heteroaryl (C? -C9) oxyalkyl (C? -Ce) aryl of C6-C? 0, aryl (C6-C? o) oxyalkyl (C C6) heterocycle of C1-C9, heteroaryl (C? -9) oxyalkyl (Cr C6) heteroaryl of C Cg, aryl (C6-C1o) apl (C6-C? o) alkyl of C? -C6, heteroaryl (Cr C9) aryl (C6-C? o) alkyl of CrC6, aryl (C6-C? o) heteroaryl] (C? -C9) C? -C6 alkyl, heteroaryl (C? -C9) heteroaryl (C? -C9) C? -C6 alkyl, aryl (C6-C? O) (C? -C6) alkoxyC? -C6 alkyl, and heteroaryl (C? -C9) (C? -C6) alkoxyC6 alkyl, in the that, independently, each of the ring carbon atoms of said C6-C? 0 aryl moieties and d-Cg heteroaryl which is capable of forming an additional bond, is optionally substituted with a selected group between fluorine, chlorine, bromine, CI-CT alkyl, Ci-Cß alkoxy, perfluoroalkyl d-C3 and perfluoroalkoxy of C? -C3. R2 and R3 are selected, each independently, between hydrogen and C? -C6 alkyl, or taken together form a ring of a spiro compound, of the formula (ring linkage) (CH2) /; (CH2) m X where X is a bond, CH2, O, S, NH or N-C? -C6 alkyl, n is independently 1 or 2, and m is independently 1 or 2; Y R4 is hydrogen or Ci-Cß alkyl- In a preferred aspect of the invention, R is selected from group consisting of C6-C? 0 aryl, C1-C9 heteroaryl, aryl (C6-C? o) oxyaryl of Ce-C or, heteroaryl (C? -Cg) oxyallo of C6-C? o, aryl (C6-C? 0) aryl of Ce-Cío, heteroaryl (CrCg) of C6-C? 0, aryl (C6-C? o) alkoxy (C? -C6) aryl of C6-C? 0, and heteroaryl (C? -Cg) C 6 -C 6 alkoxy C 6 -C 0 aryl. In further preferred aspects of the invention, R1 is selected: (a) from the group consisting of 4- [aryl (C6-C? O)] phenyl, 4- [aryl (C6-C? 0) oxy] phenyl and 4- [aryl (C6-C? O) alkoxy (C? -C6) phenyl; or (b) between the group consisting of 4- [heteroaryl (CrCg)] phenyl, 4- [heteroaryl (C? -8) oxy] phenyl and 4- [heteroaryl (C? -Cg) alkoxy (C? -C6 )]phenyl. Extremely preferred examples include those in which R1 is 4- (4-fluorophenoxy) phenyl, 4- (4-chlorophenoxy) phenyl and 4- (naphthalene-2-yloxy) phenyl. In a preferred aspect of the invention R2 and R3, taken together, form a ring of a spiro compound, of the formula: (ring attachment) wherein X is a bond, CH2, O, S, NH or N-Ci-Cß alkyl, n is 1 or 2, and m is 1 or 2, so that n is the same as m. In a further aspect of the preferred invention, R 2 and R 3 are selected from hydrogen and C 1 -C 6 alkyl. According to this aspect of the invention, it is preferred that R 2 is the same as R 3, ie, R 2 and R 3 are each hydrogen, or are each C 1 -C 6 alkyl, so that R 2 is the same than R3. In preferred examples of the invention R 4 is C 1 -C 6 alkyl; R4 is hydrogen; and R2, R3 and R4 are each hydrogen. In a highly preferred embodiment of the invention, the ring carbon to which R4 is attached has the configuration R. Accordingly, preferred compounds of the invention are provided according to formula (I ') wherein the preference in the selection of the groups R1, R2, R3 and R4 is as mentioned above with respect to compounds in which the stereospecific configuration is unspecified in the carbon atom of the ring to which R4 binds. Thus, preferred compounds of the invention include: (4r?) -1- [4- (4-fluorophenoxy) benzyl] -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4R) -1- [4-naphthalen-1-yloxy) benzyl] -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4ft) -1- [4-naphthalen-2-yloxy) benzyl] -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4f?) - 1- (4-methoxybenzyl) -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4R) -1- [3- (4-fluorophenoxy) benzyl] -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4R?) - 1-naphthalen-2-methyl-2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4R) -1- (4'-Fluorobiphenyl-4-ylmethyl) -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4) -1- (4-benzyloxybenzyl) -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; and (4A?) - 1- [4- (2-Chloro-4-fluorobenzyloxy) -benzyl] -2-oxo-imidazoline-4-carboxylic acid hydroxyamide. Additional preferred compounds of the invention include: (4R) -1- [4- (4-chlorophenoxy) benzyl] -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; hydroxyamide of (4R) -1- [4- (4-fluorophenoxy) benzyl] -2-oxo-7-oxa-1,3-diazaspiro [4,4] nonane-4-carboxylic acid; (4R) -2-oxo-1- [4- (pyrimidin-4-yloxy) benzyl] -imidazolidine-4-carboxylic acid hydroxyamide; (4 ??) -4-methyl-2-oxo-1- [4- (pyridin-4-yloxy) benzyl] imidazolidine-4-carboxylic acid hydroxyamide; (4R?) - 5,5-dimethyl-2-oxo-1- [4- (pyrimidin-4-yloxy) -benzyl] imidazolidine-4-carboxylic acid hydroxyamide; (4) -1- [4- (4-fluorophenoxy) benzyl] -5,5-dimethyl-2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4R) -1- [4- (4-chlorophenoxy) benzyl] -5,5-dimethyl-2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; hydroxyamide of (4R) -1- [4- (4-fluorophenoxy) benzyl] -4-methyl-2-oxo-imidazolidine-4-carboxylic acid; (4R) -1- [4- (4-chlorophenoxy) benzyl] -4-methyl-2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; hydroxyamide of (4R) -1- [4- (4-fluorophenoxy) benzyl] -2-oxo-1,3-diazaspiro [4,4] nonane-4-carboxylic acid; (4?) - 1- [4- (4-chlorophenoxy) benzyl] -2-oxo-1,3-diazaspiro [4,4] nonane-4-carboxylic acid hydroxyamide; (4R) -1- [4- (4-fluorophenoxy) benzyl] -4,5,5-t-methyl-2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4R) -1- [4- (4-chlorophenoxy) benzyl] -4,5,5-trimethyl-2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4R) -4-methyl-1- [4- (naphthalen-2-yloxy) -benzyl] -2-oxo-imidazol-4-carboxylic acid hydroxyamide; (4ft) -5,5-dimethyl-1- [4- (naphthalen-2-yloxy) benzyl] -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4R) -1- [4- (5-fluoropyridin-2-yloxy) benzyl] -4-methyl-2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4 /?) - 2-Oxo-1- (4-pyridin-4-ylbenzyl) imidazolidine-4-carboxylic acid hydroxyamide; and (4) -2-oxo-1- (4-pyridylmethyl) -imidazole-4-carboxylic acid hydroxyamide. The term "alkyl" as used herein, unless otherwise indicated, includes saturated monovalent hydrocarbon radicals having straight, branched or cyclic moieties or combinations thereof. The term "alkoxy" as used herein, includes O-alkyl groups, wherein "alkyl" is as defined above. The term "aryl" as used herein, unless otherwise indicated, includes an organic radical derived from a monocyclic or bicyclic C 6 -C 6 aromatic hydrocarbon, by removal of a hydrogen, such as phenyl or naphthyl, optionally substituted with substituents selected from the group consisting of fluoro, chloro, bromo, perfluoroalkyl of C-Cβ (including trifluoromethyl), C-Cß alkoxy, C3-perfluoroalkoxy (including trifluoromethoxy and difluoromethoxy) and CrC6 alkyl. Unless indicated otherwise, the selection of each of the optional substituents is independent of the selection of any other optional substituents, and preferably, the number of substituents is zero, or is between 1 and 3. The term "heteroaryl" as used herein, unless otherwise indicated, includes an organic radical derived from an aromatic heterocyclic compound of C1-C9, monocyclic or bicyclic, by separation of a hydrogen, such as pyridyl, furyl, pyrrolyl, thienyl, isothiazolyl, imidazolyl, benzimidazolyl, tetrazolyl, pyrazinyl, pyrimidyl, quinolyl, isoquininoyl, benzofuryl, isobenzofuryl, benzothienyl, pyrazolyl, indolyl, isoindolyl, purinyl, carbazolyl, isoxazolyl, thiazolyl, oxazolyl, benzothiazolyl and benzoxazolyl, optionally substituted with substituents selected from the group consisting of fluoro, chloro, trifluoromethyl, C1-C6 alkoxy, trifluoromethoxy, difluoromethoxy and Ci-Cß alkyl. Unless otherwise indicated, the selection of each optional substituent is independent of the selection of any other optional substituents and, preferably, the number of substituents is zero, or is between 1 and 2. "A suitable substituent" is understood which means a chemically and pharmaceutically acceptable functional group, that is, a moiety that does not substantially negate the inhibitory activity of the compounds of the invention, and / or a moiety that provides useful properties to the production, storage or use of the compounds of the invention. invention as pharmaceutical compounds. Such suitable substituents can be determined by those skilled in the art. Illustrative examples of suitable substituents include, but are not limited to, alkyl groups, hydroxy groups, alkylthio groups, alkoxy groups, carboxy groups, amino groups, alkyl- and dialkylamino groups, carbamoyl groups, alkylcarbonyl groups, alkoxycarbonyl groups, groups alkylaminocarbonyl, dialkylaminocarbonyl groups, arylcarbonyl groups, aryloxycarbonyl groups, alkylsulfonyl groups, arylsulfonyl groups and the like. The compound of formula I can have chiral centers, and therefore it can exist in different enantiomeric forms. This invention relates to all optical isomers, tautomers and stereoisomers of the compounds of formula I and mixtures thereof.

The present invention also relates to the pharmaceutically acceptable acid addition salts of the compounds of formula I. The acids which are used to prepare the pharmaceutically acceptable acid addition salts of the aforementioned basic compounds of this invention, are those which form non-toxic acid addition salts, ie, salts containing pharmacologically acceptable anions, such as the hydrochloride, hydrobromide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, acetate, lactate, citrate, acid citrate, tartrate, bitartrate salts , succinate, maleate, fumarate, gluconate, saccharate, benzonate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate [i.e. 1, 1'-methylene-bs (2-hydroxy-3-naphthoate)]. The invention also relates to base addition salts of formula I. Chemical bases which can be used as reagents for preparing pharmaceutically acceptable base salts of the compounds of formula I having an acid nature, are those which form salts with such compounds non-toxic base Such non-toxic base salts include, but are not limited to, those derived from pharmacologically acceptable cations, such as alkali metal cations (e.g., potassium and sodium), and alkaline earth metal cations (e.g., calcium and magnesium) , ammonium or water-soluble amine addition salts, such as N-methylglucamine- (meglumine), and the salts of lower alkanol ammonium and other base salts of pharmaceutically acceptable organic amines.

The subject invention also includes isotopically-labeled compounds, which are identical to those cited in formula I, except the fact that one or more atoms are replaced by an atom having an atomic mass or a mass number different from the atomic mass or the mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as 2H, 3H, 13C, 14C, 15N, 180, 170, 31P, 32P, 35S , 18F, 36CI, respectively. The compounds of the present invention, their prodrugs and the pharmaceutically acceptable salts of said compounds or of said prodrugs which contain the aforementioned isotopes and / or other isotopes or other atoms, are within the scope of this invention. Certain isotopically-labeled compounds of the invention, for example, those in which radioactive isotopes such as 3H and 14C are incorporated, are useful in the performance of drug distribution and / or substrate tissue distribution assays. Tritiated, ie, 3H, and carbon-14, i.e. 14C, isotopes are particularly preferred for their ease of preparation or ability to detect. In addition, replacement with heavier isotopes such as deuterium, i.e., 2H, may provide certain therapeutic advantages resulting from increased metabolic stability, eg, increased in vivo half-life or reduced dosage requirements and, therefore, may be preferred in certain circumstances. The compounds of formula I of this invention and their prodrugs, isotopically labeled, can be prepared, in general, by carrying out the operating procedures described in the schemes and / or in the examples and preparations listed below, by substituting an unlabelled reagent. isotopically by an isotopically-labeled reagent, readily available. The present invention also relates to a pharmaceutical composition for the treatment of a condition selected from the group consisting of arthritis (including osteoarthritis and rheumatoid arthritis), inflammatory bowel disease, Crohn's disease, emphysema, respiratory insufficiency syndrome acute, asthma, chronic obstructive pulmonary disease, Alzheimer's disease, organ transplant toxicity, cachexia, allergic reactions, allergic contact hypersensitivity, cancer (such as cancers of solid tumors including colon cancer, breast cancer, lung cancer and prostate cancer) and hematopoietic malignancies, including leukemias and lymphomas, tissue ulceration, restenosis, periodontal disease, epidermolysis bullosa, osteoporosis, loosening of implants of artificial joints, atherosclerosis (including rupture of atherosclerotic plaques), aortic aneurysm (including abdominal aortic aneurysm and cerebral aortic aneurysm) congestive heart failure, myocardial infarction, stroke, ischemia cerebral, head trauma, spinal cord damage, neurodegenerative disorders (acute and chronic), autoimmune disorders, Huntington's disease, Parkinson's disease, migraines, depressions, peripheral neuropathies, pain, cerebral amyloid angiopathy, nootropic or knowledge improvement , scleros is amyotrophic lateral, multiple sclerosis, ocular angiogenesis, corneal damage, macular degeneration, abnormal healing of wounds, burns, diabetes, tumor invasion, tumor growth, tumor metastasis, corneal scars, scleritis, AIDS and sepsis and septic shock in a mammal , including the human being, comprising an amount of a compound of formula I or a pharmaceutically acceptable salt thereof, effective in such treatments, and a pharmaceutically acceptable excipient. The present invention also relates to a pharmaceutical composition for the treatment of diseases characterized by metalloproteinase activity (preferably MMP-13) and other diseases characterized by reprolysin activity in mammals (preferably Agrecanase activity and most preferably Agrecanase activity) in a mammal, including the human, comprising an amount of a compound of formula I or a pharmaceutically acceptable salt thereof effective in such treatments and a pharmaceutically acceptable excipient. The present invention also relates to a pharmaceutical composition for the inhibition of (a) matrix metalloproteinases or other metalloproteinases involved in the degradation of the matrix, or (b) a mammalian reprolysin (such as aggrecanase or ADAMs TS-1, 10, 12, 15, and 17, most preferably Agrecanase) in a mammal, including human, comprising an effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. The present invention also relates to a method for treating a condition selected from the group consisting of arthritis (including osteoarthritis and rheumatoid arthritis), inflammatory bowel disease, Crohn's disease, emphysema, acute respiratory distress syndrome, asthma, obstructive pulmonary disease chronic, Alzheimer's disease, organ transplant toxicity, cachexia, allergic reactions, allergic contact hypersensitivity, cancer (such as cancers of solid tumors including colon cancer, breast cancer, lung cancer and prostate cancer) and haematopoietic malignancies (including leukemias and lymphomas), tissue ulceration, restenosis, periodontal disease, epidermolysis bullosa, osteoporosis, loosening of implants of artificial joints, atherosclerosis (including rupture of atherosclerotic plaques) aortic aneurysm (including abdominal aortic aneurysm and aortic aneurysm) cerebral ico), congestive heart failure, myocardial infarction, stroke, cerebral ischemia, head trauma, spinal cord damage, neurodegenerative disorders (acute and chronic), autoimmune disorders, Huntington's disease, Parkinson's disease, migraines , depressions, peripheral neuropathies, pain, cerebral amyloid angiopathy, nootropic or knowledge improvement, amyotrophic lateral sclerosis, multiple sclerosis, ocular angiogenesis, corneal damage, macular degeneration, abnormal healing of wounds, burns, diabetes, tumor invasion, tumor growth, tumor metastasis, corneal scars, scleritis, AIDS, and sepsis and septic shock in a mammal, including humans, comprising administering to said mammal an amount of a compound of formula I or a pharmaceutically acceptable salt thereof, effective to treat such a condition. The present invention also relates to the treatment of diseases characterized by matrix metalloproteinase activity (preferably MMP-13 activity) and other diseases characterized by mammalian reprolysin activity (preferably Agrecanase activity) in a mammal, including a human, which comprises administering to said mammal an amount of a compound of formula I or a pharmaceutically acceptable salt thereof, effective to treat such a condition. The present invention also relates to a method for the inhibition of (a) matrix metalloproteinases or other metalloproteinases involved in the degradation of the matrix, or (b) a mammalian reprolysin (such as aggrecanase or TS-1, 10, 12, 15, 17 of ADAM, preferably Agrecanase) in a mammal, including a human, which comprises administering to said mammal an effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof. The present invention also relates to a method for inhibiting the cleavage of TNF-α from cell membranes in a mammal, which comprises administering to said mammal an effective amount of a compound of formula I which inhibits Agrecanase. The present invention also relates to a method of treatment of arthritis in a mammal, which comprises administering to said mammal an effective amount of an inhibitor of Agrecanase, wherein said inhibitor of Agrecanase selectively inhibits Agrecanase in preference to MMP. -1. The present invention also relates to a method of treating arthritis in a mammal, comprising administering to said mammal an effective amount of an inhibitor of Agrecanase, wherein said inhibitor of Agrecanase selectively inhibits aggrecanase at least ten. times more than MMP-1. The present invention also relates to a method of treating arthritis in a mammal, which comprises administering to said mammal an effective amount of an inhibitor of Agrecanase, wherein said inhibitor of Agrecanase selectively inhibits Agrecanase and the MMP-13 in preference to MMP-1. The present invention also relates to a method of treating arthritis in a mammal, which comprises administering to an mammal such an effective amount of an inhibitor of the Agrecanase, wherein said inhibitor of the Agrecanase selectively inhibits the Agrecanase and the MMP-13, at least ten times more than MMP-1.

The present invention also relates to a method of treating arthritis in a mammal, which comprises administering to said mammal an effective amount of an inhibitor of Agrecanase of a hydroxamic acid, wherein said inhibitor of Agrecanase selectively inhibits the Agrecanase and the MMP-13 in preference to MMP-1. The present invention also relates to a method of treating arthritis in a mammal, which comprises administering to said mammal an effective amount of an inhibitor of Agrecanase of a hydroxamic acid, wherein said inhibitor of Agrecanase selectively inhibits the Agrecanase and the MMP-13 at least ten times more than MMP-1. The term "treat", as used herein, refers to investment, alleviation, inhibition of progress, or prevention of the disorder or condition to which such a term applies, or one or more symptoms of such disorder or condition. The term "treatment", as used herein, refers to the act of treating, as "treating" has been defined immediately before. This invention also includes pharmaceutical compositions containing prodrugs of compounds of formula I. This invention also includes methods of treating or preventing disorders that can be treated or prevented by inhibiting matrix metalloproteinases or inhibiting mammalian reprolysin, which it comprises administering prodrugs of compounds of formula I. Compounds of formula I that possess free amino, amido, hydroxy or carboxylic groups can be converted into prodrugs. Prodrugs include compounds in which a residue of an amino acid, or a chain of a polypeptide of two or more (eg, two, three or four) amino acid residues are covalently linked by peptide linkages to amino, hydroxy or carboxylic acid groups free of compounds of formula I. The amino acid residues include the 20 natural amino acids commonly designated by three-letter symbols and also include, 4-hydroxyproline, hydroxylysine, demosin, isodemosin, 3-methylhistidine, norvaline, beta-alanine, gamma-acid. aminobutyric acid, citrulline, homocysteine, homoserin, ornithine and methionine sulfone. The prodrugs also include compounds in which carbonates, carbamates, amides and alkyl esters of which are covalently linked to the substituents of formula I above by the carbonyl carbon of the side chain of the prodrug. Those skilled in the art will appreciate that the compounds of the invention are useful for treating a diverse set of diseases. Those skilled in the art will also appreciate that when the compounds of the invention are employed in the treatment of a specific disease, the compounds of the invention may be combined with various existing therapeutic agents employed for such a disease. For the treatment of rheumatoid arthritis, the compounds of the invention can be combined with agents such as TACE inhibitors, TNA-a inhibitors such as anti-TNF monoclonal antibodies and TNF receptor immunoglobulin molecules (such as Enbrel®) , COX-2 inhibitors, low-dose methotrexate, lefunimide, hydroxychloroquine, d-penicillamine, auranofin or other parenteral or oral. The compounds of the invention can also be used in combination with existing therapeutic agents for the treatment of osteoarthritis. Suitable agents to be used in combination include non-spheroidal anti-inflammatory agents, standard (hereinafter NSAIDs) such as piroxicam, diclofenacpropionic acids such as naproxen, flubiprofen, fenoprofen, ketoprofen and ibuprofen, fenamates such as mefenamic acid, indomethacin, sulindac, apazone, pyrazolones such as phenylbutazone, salicylates such as aspirin, COX-2 inhibitors such as celecoxib and rofecoxib, analgesics and intraarticular therapeutic agents such as corticosteroids and hyaluronic acids such as hyalgan and sinvisc. The compounds of the present invention can also be used in combination with anticancer agents such as endostatin and angiostatin or cytotoxic drugs such as adriamycin, daunomycin, cis-platinum, etoposide, taxol, taxotere and alkaloids such as vincristine, and antimetabolites such as methotrexate. The compounds of the present invention can also be used in combination with cardiovascular agents such as calcium channel blockers, lipid lowering agents, such as statins, fibrates, beta-blockers, Ace inhibitors, Angiotensin receptor antagonists. 2 and inhibitors of platelet aggregation.

The compounds of the present invention can also be used in combination with CNS agents such as antidepressants (such as sertraline), antiparkinsonian drugs (such as deprenyl, L-dopa, requib, miratex, MAOB inhibitors such as selegine and rasagiline, inhibitors of comP such as Tasmar, A-2 inhibitors, dopamine reuptake inhibitors, NMDA antagonists, nicotine agonists, dopamine agonists and neuronal nitric oxide synthase inhibitors), anti-Alzheimer drugs such as donepezil, tacrine, COX inhibitors -2, propentofylline or metrifonate. The compounds of the present invention can also be used in combination with anti-osteoporosis agents such as roloxifene, droloxifene or fosomax, and immunosuppressive agents such as FK-506 and rapamycin.

DETAILED DESCRIPTION OF THE INVENTION The reaction schemes that follow illustrate the preparation of the compounds of the present invention. Unless otherwise indicated, R1, R2, and R3, and R4 in the reaction schemes and the discussion that follows, are defined as above.

SCHEME! ((li) General reaction conditions With reference to scheme 1, the compounds of the formula I can be prepared from compounds of the formula II by removal of the protective group of the hydroxyamide P2, which can be P2 tere-butyl, benzyl, -trimethylsilylethyl or allyl The preferred protecting group is 2-trimethylsilylethyl When P2 is benzyl, the separation of the protective group from the hydroxyamide is carried out by hydrogenolysis using catalytic palladium on barium sulfate, in a polar solvent such as methanol, at a temperature of about 20 ° C. When P2 is 2-trimethylsilylethyl, removal of the protecting group from the hydroxyamide is carried out using boron trifluoride etherate in an inert solvent such as methylene chloride or chloroform, preferably methylene chloride , at a temperature from about 0 ° C to about 50 ° C, preferably about 20 ° C. When P3 is tere-butyl, the separation of the protecting group is carried out using a strong acid such as trifluoroacetic acid in an inert solvent such as methylene chloride or chloroform, preferably methylene chloride, at a temperature from about 0 ° C to about 50 ° C, preferably 20 ° C approximately. When P2 is allyl, the removal of the protecting group can be carried out by treatment with tributyltin hydride and acetic acid in the presence of catalytic bis (triphenylphosphine) palladium (II) chloride. With reference to scheme 1, the compounds of the formula II can be prepared from carboxylic acids of the formula III by reaction with a hydroxylamine derivative of the formula P2ONH2 in the presence of an activating agent such as 1- (3-dimethylaminopropyl) 3-ethylcarbodiimide and 1-hydroxybenzotriazole in an aprotic solvent, such as methylene chloride or N, N-dimethylformamide, preferably methylene chloride. The reaction is carried out at a temperature of about 0 ° C to about 50 ° C, preferably about 20 ° C. The hydroxylamine of the formula P2ONH2 is preferably generated in situ starting from a salt form, such as hydrochloride, in the presence of a base, such as triethylamine or the diisopropylethylamine, preferably diisopropylethylamine. The compounds of the formula III can be prepared from compounds of the formula IV by removal of the protective group P1 from the carboxylic acid, where P1 is methyl, ethyl or tere-butyl, preferably tere-butyl. When P1 is methyl or ethyl, the removal of the protecting group P1 is carried out by reaction with excess of a hydroxide of a metal, such as sodium hydroxide or lithium hydroxide, preferably lithium hydroxide, in a protic solvent, such as aqueous ethanol, at a temperature from about 0 ° C to about 100 ° C, preferably about 20 ° C. In cases where the solubility of IV is limited, tetrahydrofuran may be added to the reaction mixture as a complementary solvent. When p1 is tere-butyl, the separation of the protecting group p1 is carried out by treatment with a strong acid such as hydrochloric acid or trifluoroacetic acid, preferably trifluoroacetic acid, in an inert solvent such as chloroform or methylene chloride, preferably methylene chloride. The reaction is carried out at a temperature from about 0 ° C to about 50 ° C, preferably about 20 ° C. The compounds of formula IV can be prepared from compounds of formula V by hydrogenation under hydrogen atmosphere, in the presence of a catalyst, in a solvent inert to the reaction. Suitable catalysts include palladium on carbon, palladium on carbon hydroxide or palladium black, preferably palladium on carbon. Suitable solvents include an alcohol such as ethanol or methanol, preferably methanol. The aforesaid reaction can be carried out at a pressure of from about 1 to about 5 atmospheres, preferably about 3 atmospheres. Suitable temperatures for the aforesaid reaction range from about 20 ° C (room temperature) to about 60 ° C, preferably about 20 ° C. The compounds of formula V can be prepared from the compounds of formula VI by reaction with a base and an alkylating agent of formula R1 (CH2) -X, wherein X is a removable group such as Br, I or toluenesulfonate. Suitable bases include potassium carbonate, cesium carbonate, potassium hexamethyldisilazide, or sodium hydride, preferably potassium carbonate. The reaction mixture is stirred in a polar solvent such as acetone, N, N-dimethylformamide, or N-methylpyrrolidin-2-one, at a temperature from about 0 ° C to about 50 ° C, preferably about 20 ° C.

The compounds of formula V in which R 4 is C C β alkyl can be obtained by alkylation of compounds of formula V in which R 4 is hydrogen. The alkylation is carried out by reaction of an intermediate of formula V in which R 4 is hydrogen, with an alkyl halide of the formula CH 3 (CH 2) nX wherein n is 0 to 5 and X is bromine or iodine. The aforesaid reaction is carried out in the presence of a strong base with spherical hindrance such as lithium diisopropylamide or lithium hexamethyldisilazide in an inert solvent such as diethyl ether or tetrahydrofuran, at a temperature from about -78 ° C to about 0 ° C, preferably -78 ° C approximately. Compounds of formula VI can be prepared from compounds of formula VII by reaction with benzyl chloroformate, in the presence of a base such as triethylamine or diisopropylethylamine, preferably triethylamine, and a catalytic amount of 4-dimethylaminopyridine. The aforesaid reaction is carried out in a solvent such as tetrahydrofuran, methylene chloride or chloroform, preferably methylene chloride, at a temperature from about 0 ° C to about 20 ° C, preferably about 20 ° C. The compounds of the formula VII can be obtained from diamino compounds of the formula VIII by reaction with phosgene, carbonyldiimidazole or triphosgene, preferably triphosgene, in the presence of a base such as pyridine or triethylamine, preferably triethylamine. The aforesaid reaction is carried out in a solvent such as tetrahydrofuran, methylene chloride or chloroform, preferably tetrahydrofuran, at a temperature from about 0 ° C to about 20 ° C, preferably about 20 ° C. The compounds of formula VIII in which R 1 is methyl or ethyl, R 4 is hydrogen and R 2 and R 3 are independently C 1 -C 6 alkyl, can be obtained from ketones of the formula R 2 R 3 CO wherein R 2 and R 3 are , independently, C -CT alkyl. Similarly, compounds of formula VIII in which p1 is methyl or ethyl, R4 is hydrogen, and R2 and R3 taken together form a ring of a spiro compound of the formula wherein X is a CH2, O, S, NH or N-C6C6 alkyl bond, n is independently 1 or 2, and m is independently 1 or 2, can be prepared from cyclic ketones of the formula (IX) in which X is a bond, CH 2, O, S, NH or N-C 1 -C 2 alkyl, n is independently 1 or 2, and m is independently 1 or 2 The operative procedures are the same as those described by Schollkopf et al., In the case that R2 and R3 are methyl (Liebigs Ann. Chem. 611, 1973 and Liebigs Ann. Chem. 1183, 1977). The compounds of formula VIII wherein p 1 is methyl or ethyl, and R3 and R4 are hydrogen, they can be prepared from compounds of the formula X: wherein R3 is C-C6 alkyl. The operative procedures are the same as those described by Mohán et al., In case R3 is isopropyl (J. Med. Chem. 34, 2402, 1991). Various methods of preparing compounds of formula IX are known in the scientific literature, for example Shin et al., Bull. Chem. Soc. Jpn. 45, 3595, 1972. The compound of formula VI in which P1 is tere-butyl and R2, R3 and R4 are hydrogen, is known in the scientific literature as the S-enantiomer (Shiba et al., Bull, Chem. Soc. Japan, 41, 2748, 1968). The corresponding R-enantiomer is prepared as described for the S-enantiomer using N-benzyloxycarbonyl-D-asparagine in place of N-benzyloxycarbonyl-L-asparagine as the starting material. The compounds of the formula I having a basic nature are capable of forming a wide variety of different salts with various inorganic and organic acids. Although such salts must be pharmaceutically acceptable for administration to animals, it is often desirable in practice to initially isolate a compound of the formula I from the reaction mixture in the form of a pharmaceutically unacceptable salt., then simply, converting the latter into the free base by treatment with an alkaline reagent and then converting the free base into a pharmaceutically acceptable acid addition salt. The acid addition salts of the basic compounds of this invention are readily prepared by treating the basic compound with a substantially equivalent amount of the chosen mineral or organic acid, in an aqueous solvent medium or in a suitable organic solvent, such as methanol or ethanol. By careful evaporation of the solvent, the desired solid salt is obtained. The acids which are used to prepare the pharmaceutically acceptable acid addition salts of the basic compounds of this invention are those which form non-toxic acid addition salts, ie salts containing pharmacologically acceptable anions, such as hydrochloride salts hydrobromide, hydroiodide, nitrate, sulfate or bisulfate, phosphate or acid phosphate, acetate, lactate, citrate or citrate acid, tartrate or bitartrate, succinate, maleate, fumarate, gluconate, saccharate, benzoate, methanesulfonate and pamoate [ie 1, 1 '-methylene-bs (2-hydroxy-3-naphthoate)]. Those compounds of the formula I which have an acid nature are capable of forming base salts with various pharmacologically acceptable cations. Examples of such salts include the alkali metal or alkaline earth metal salts and, in particular, the sodium and potassium salts. These salts are all prepared by conventional techniques. The chemical bases which are employed as reagents for preparing the pharmaceutically acceptable base salts of this invention, are those which form non-toxic base salts with the acidic compounds of formula I described herein. These non-toxic base salts include those derived from pharmacologically acceptable cations such as sodium, potassium, calcium and magnesium, etc. These salts can be easily prepared by treating the corresponding acidic compounds with an aqueous solution containing the desired pharmacologically acceptable cations, and then evaporating the resulting solution to dryness, preferably under reduced pressure. Alternatively, they can be prepared by mixing together solutions in lower alcohols of the acidic compounds and the desired alkali metal alkoxide, and then evaporating the resulting solution to dryness in the same manner as above. In both cases, stoichiometric amounts of reagents are preferably employed in order to ensure that the reaction is complete and the maximum product yields. To be administered to mammals, including humans, for the inhibition of matrix metalloproteinases, to inhibit the production of tumor necrosis factor (TNF) and, for example, for the inhibition of mammalian reprolysin (preferably the inhibition of aggrecanase ), a variety of a variety of conventional routes can be employed including the oral, parenteral (eg, intravenous, intramuscular or subcutaneous), buccal, anal and topical routes. In general, the compounds of the invention (hereinafter also referred to as the active compounds) will be administered in doses between about 0.1 and 25 mg / kg of weight of the subject to be treated, per day, preferably from about 0.3 to 5 mg / kg. . Preferably, the active compound will be administered orally or parenterally. However, some variation in the dose will necessarily occur, depending on the condition of the subject being treated. The person responsible for the administration will determine, in any case, the appropriate dose for the individual subject. The compounds of the present invention can be administered in a wide variety of different dosage forms; In general, the therapeutically effective compounds of this invention are present in such dosage forms at concentration levels ranging from about 5.0% to about 70% by weight. For oral administration, tablets containing various excipients such as microcrystalline cellulose, sodium citrate, calcium carbonate, dicalcium phosphate and glycine can be used, together with disintegrating agents such as starch (and preferably corn starch, potato starch or tapioca), alginic acid and certain complex silicates, together with granulation binders such as polyvinylpyrrolidone, sucrose, gelatin and gum arabic. In addition, lubricating agents such as magnesium stearate, sodium lauryl sulfate and talc are often very useful for the production of tablets. Solid compositions of a similar type can also be used as fillers in gelatin capsules; Preferred materials in this regard also include lactose or milk sugar, as well as high molecular weight polyethylene glycols. When aqueous suspensions and / or elixirs are desired for oral administration, the active ingredient may be mixed with various sweetening or flavoring agents, coloring matters, and if desired, emulsifying agents and / or suspending agent, together with diluents such as water, ethanol , propylene glycol, glycerin and various combinations thereof. In the case of animals, they are advantageously included in an animal feed or in drinking water, in a concentration of 5-5000 ppm, preferably 25 to 500 ppm. For parenteral administration (intramuscular, intraperitoneal, subcutaneous and intravenous use) a sterile injectable solution of the active ingredient is usually prepared. Solutions of a therapeutic compound of the present invention may be used either in sesame oil or in peanut oil, or in aqueous propylene glycol solution. The aqueous solutions must be adequately adjusted and buffered, preferably at a pH greater than 8, if necessary, and the liquid diluent first made isotonic. These aqueous solutions are suitable for intravenous injection purposes. Oily solutions are suitable for intra-articular, intramuscular and subcutaneous injection purposes. The preparation of all these solutions under sterile conditions is easily carried out by standard pharmaceutical techniques well known to those skilled in the art. In the case of animals, the compounds can be administered intramuscularly or subcutaneously, at dose levels of approximately 0.1 to 50 mg / kg / day, advantageously 0.2 to 10 mg / kg / day, given in a single dose or up to 3 divided doses. The active compounds of the invention can also be formulated into compositions for rectal administration, such as suppositories or retention enemas, containing, for example, conventional suppository bases such as cocoa butter or other glycerides. For intranasal administration or administration by inhalation, the active compounds of the invention are conveniently distributed in the form of a solution or suspension from a pump spray container, which is tightened or pulsed by the patient, or in the form of a spray presentation. aerosol from a pressurized container or a nebulizer, with the use of a suitable propellant agent, for example dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of an aerosol placed under pressure, the dose unit can be determined by providing a valve that distributes a measured quantity. The container placed under pressure or nebulizer may contain a solution or suspension of the active compound. Capsules and cartridges (made, for example, of gelatin) can be formulated for use in an inhaler or nsufflator, containing a powder mixture of a compound of the invention and a suitable powder base such as lactose or starch. The ability of the compounds of formula I or their pharmaceutically acceptable salts (hereinafter referred to as the compounds of the present invention) to inhibit mammalian metalloproteinases or reprolysin and, therefore, to demonstrate its efficacy in treating diseases characterized by metalloproteinases or the production of tumor necrosis factor, is evidenced by the in vitro titration assays listed below.

BIOLOGICAL TESTING Inhibition of soluble TNF production The ability of the compounds or their pharmaceutically acceptable salts to inhibit the cellular production / release of TNF and, consequently, demonstrate their efficacy in treating diseases involving TNF derangement, is evidenced by the following in vitro assay: Method for the evaluation of the activity of the recombinant TNFa conversion enzyme 1) Preparation of recombinant TACE A DNA fragment encoding the signal sequence, the prodomain and the catalytic domain of TACE (amino acids 1-473), was amplified by polymerase chain reaction using a lung cDNA library as a template human. The amplified fragment was cloned into the pFastBac vector. The DNA sequence of the insert was confirmed for both chains. A bacmid prepared using pFastBac in E. coli DHIOBac was transfected into SF9 insect cells. The virus particles were amplified to stages P1, P2, P3. The F3 virus was infected in Sf9 and High Five insect cells and cultured at 27 ° C for 48 hours. The medium was collected and used for the tests and their subsequent purification. 2) Preparation of fluorescent substrate off A model peptide TNF-α substrate (LY-Leucine-Alanine-Glutamine-Alanine-Valine-Arginine-Serine-Serine-Lysine (CMTR) - Arginine (LY = Lucifer Yellow; CMTR = 5-carboxyltetramethyl Rodamine)) and the concentration was estimated by absorbance at 560 nm (E56o, 60,000 M-1CM-1) according to the method of Geoghegan, KF, "Improved method for converting an unmodified peptide to an energy-transfer substrate for to proteinase. " Bioconiuqate Che, 7, 385-391 (1995). This peptide includes the cleavage site on TNF that is split in vivo by TACE. 3) Enzymatic reaction The reaction, carried out in a 96-well plate (Dynatech), was composed of 70 μl of buffer solution (25 mM Hepes-HCl, pH 7.5, plus 20 uM ZnCl 2), 10 μl of 100 μM quenched fluorescent substrate, 10 μl of a test compound solution in DMSO (5 μM). %), and an amount of r-TACE enzyme that can cause a 50% excision in 60 minutes, in a total volume of 100 μl. The specificity of the cleavage by the enzyme in the amide link between alanine and valine was verified by HPLC and mass spectrometry. The initial rates of excision were verified by measuring the degree of increase in fluorescence at 530 nm (excitation at 409 nm) over 30 minutes. The experiment was controlled as follows: 1) by the background fluorescence of the substrate; 2) by the fluorescence of the fully cleaved substrate; 3) by the depletion or increase of fluorescence from solutions containing the test compound. The results obtained were analyzed as follows. Speeds from "control" reactions that did not contain test compound were averaged to establish the 100% value. The reaction rate in the presence of test compound was compared to that obtained in the absence of compound, and tabulated as "percent of the control that did not contain test compounds". The results were graphically represented as "% of the control" against the log. Of the concentration of compounds and the value in the semi-maximum point or IC50 was determined. The IC 50 for the above test is a measure of the inhibition of the proteolytic activity of TACE on TNF-α. The blocking of the binding of TNF-a to TACE as used herein is in accordance with that described in the patent of E.U.A. No. 5,830,742, issued November 3, 1998.

Monocyte analysis Human mononuclear cells were isolated starting from anti-coagulated human blood, using a Ficoll-hypaque single-step separation technique. (2) the mononuclear cells were washed three times with Hanks Balanced Salt Solution (HBSS) with divalent cations and resuspended at a density of 2 x 106 / ml in HBSS containing 1% BSA. The differential counts determined using the Abbott Cell Dyn 3500 analyzer indicated that monocytes varied in these preparations from 17 to 24% of the total cells. 180 m of the cell suspension was aliquoted into 96-well flat bottom plates (Costar). Additions of compounds and LPS (100 ng / ml final concentration) gave a final volume of 200 μl. All the conditions were carried out in triplicate. After an incubation period of four hours at 37 ° C in an incubator with C02, humidified, the plates were removed and subjected to centrifugation (10 minutes at approximately 250 x g) and the supernatants were separated and analyzed for TNF-a using the ELISA kit of R and D.

MMP assays Selective inhibitors of collagenase-3 (matrix metalloproteinase-13) as used in this specification, allude to agents that demonstrate at least 100-fold greater selectivity for the inhibition of the enzyme activity of the collagenase-3 relative to the enzymatic activity of collagenase-1 and a potency of less than 100 nM as defined by the IC50 results of the fluorescence assays of MMP-13 / MMP-1 described below. Selective inhibitors of collagenase-3 can be identified by screening the inhibitors of the present invention by the fluorescence assays of MMP-13 / MMP-1 which are described below and selecting those agents with IC50 inhibition ratios of MMP-13 / MMP-1 of 100 or more and a power of less than 100 nM. Non-selective collagenase inhibitors, as used in this specification, allude to agents that demonstrate activity less than 100 times more for the inhibition of the enzymatic activity of collagenase-3 on the enzymatic activity of collagenase-1 or a potency of more than 100 nM, as defined by the IC50 results obtained from the fluorescence assays of MMP-13 / MMP-1 described below. The ability of collagenase inhibitors to inhibit collagenase activity is well known in the art. The following tests can be used to identify inhibitors of matrix metalloproteinases.

Inhibition of human colaqenase (MMP-1) Recombinant human collagenase is activated with trypsin using the following ratio: 10 μg of trypsin per 100 μg of collagenase. Trypsin and collagenase are incubated at room temperature for 10 minutes and then a quintuple excess (50 μg / 10 μg trypsin) of soybean trypsin inhibitor is added. 10 mM stock solutions of inhibitors are prepared in dimethyl sulfoxide and then diluted using the following scheme: 10 mM? 120 μM? 12 μM? 1.2 μM? 0.12 μM Twenty-five microliters of each of the concentrations are then added in triplicate to appropriate wells of a 96-well microfluor plate. The final inhibitor concentration will be a 1: 4 dilution after the addition of enzyme and substrate. Positive controls (enzyme, without inhibitor) are established in wells D1-D6 and wells (without enzyme or inhibitors) in wells D7-D12.

Collagenase is diluted to 400 ng / ml and then 25 μl is added to the appropriate wells of the microfluor plate. The final concentration of collagenase in the assay is 100 ng / ml. The substrate (DNP-Pro-Cha-Gly-Cys (Me) -His-Ala-Lys (NMA) -NH2) is prepared in the form of a 5 mM stock solution in dimethyl sulfoxide and then diluted to 20 mM in assay buffer. The assay is initiated by the addition of 50 μl of substrate per well of the microfluor plate obtaining a final concentration of 10 μM. Fluorescence readings (excitation wavelength 360 nm, emission 460 nm) were carried out at time 0 and then at 20 minute intervals. The test is carried out at room temperature with a typical test time of 3 hours. The fluorescence is then plotted against time for both the blank and the samples containing collagenase (the average of the results of determinations made in triplicate is obtained). A time point is chosen that provides a good signal (the target) and what is on the linear part of the curve (usually around 120 minutes) to determine the IC 50 values. Zero time is used as a blank for each of the compounds in each concentration and these values are subtracted from the results obtained at 120 minutes. The results obtained are plotted as concentration of inhibitor against% of the control (fluorescence of the inhibitor divided by fluorescence of collagenase alone x 100). IC50's are determined from the concentration of inhibitor that provides a signal that is 50% of that of the control. If it is indicated that the IC5o are < 0.03 μM, then the inhibitors are tested at concentrations of 0.3 μM, 0.03 μM, 0.03 μM and 0.003 μM.

Inhibition of gelatinase (MMP-2) Inhibition of gelatinase activity is assayed using the substrate Dnp-Pro-Cha-Gly-Cys (Me) -His-Ala-Lys (NMA) -NH2 (10 μM) in them conditions that inhibit human collagenase (MMP-1). 72 kD Gelatinase is activated with 1 mM APMA (papillofenilnercuric acetate) for 15 hours at 4 ° C and diluted to obtain a final concentration in the 100 mg / ml assay. The inhibitors are diluted as for the inhibition of human collagenase (MMP-1) to give final concentrations in the assay of 30 μM, 3 μM, 0.3 μM and 0.03 μM. Each concentration is done in triplicate. Fluorescence readings (excitation wavelength 360 nm, emission of 460 nm) are taken at zero time and then at 20 minute intervals for 4 hours. The IC 50 are determined as for the inhibition of human collagenase (MMP-1). If it is indicated that the IC50's are lower than 0.03 μM, the inhibitors are tested at final concentrations of 0.3 μM, 0.03 μM, 0.003 μM and 0.003 μM.

Inhibition of stromelysin activity (MMP-3) The inhibition of stromelysin activity is based on a modified spectrophotometric assay described by Weingarten and Feder (Weingarten, H. and Feder, J., Spectrophotometric Assay for Vertébrate Collagenase, Anal. Biochem 147, 437-440 (1985)). Hydrolysis of thiopeptolide [Ac-Pro-Leu-Gly-SCH [CH2CH (CH3) 2] CO-Leu-Gly-OC2H5] which serves as a substrate provides a mercaptan fragment which can be observed in the presence of Ellman's reagent. Recombinant human proestromelysin is activated with trypsin using a ratio of 1 μl of a stock solution of tripisine of 10 mg / ml per 26 mg of stromelysin. Trypsin and stromelysin are incubated at 37 ° C for 15 minutes, followed by 10 μl of 10 μg / ml soybean trypsin inhibitor solution, for 10 minutes, at 37 ° C, to quench the trypsin activity. The assays are carried out in a total volume of 250 ml of assay buffer (200 mM sodium chloride, 50 mM MES and calcium chloride mM, pH 6.0) in 96-well microtiter plates. The activated stromelysin is diluted in assay buffer at 25 μg / ml. The reagent is prepared Ellman (3-carboxy-4-nitrophenyl disulfide) in the form of a stock solution (stock) 1 M in dimethylformamide and diluted to 5 mM in assay buffer with 50 ml per well obtaining a final concentration of 1 mM. 10 mM stock solutions of dimethyl sulfoxide inhibitors are prepared and serially diluted with assay buffer such that addition of 50 μl to the appropriate wells yields a final concentration of 3 μM, 0.3 μM, 0.003 μM and 0 μM. , 0003 μM. All conditions are completed in triplicate. A 300 mM stock solution, in dimethyl sulfoxide, of the peptide serving as substrate, is diluted to 15 mM with assay buffer and the assay is initiated by the addition of 50 μl to each well to give a final concentration of substrate. mM. The targets are constituted by the peptide that serves as Ellman's substrate and reagent, without the enzyme. The product formation was verified at 405 nm with a UVmax plate reader from Molecular Devices. The IC50 values were determined in the same way as for collagenase.

Inhibition of MMP-13 Recombinant human MMP-13 is activated with 2 mM APMA (p-aminophenylmercuric acetate) for 1.5 hours at 37 ° C and diluted to 400 mg / ml in assay buffer (50 mM Tris, pH 7.5, 200 mM sodium chloride, 5 mM calcium chloride, zinc chloride 20 μM, Brij at 0.02%). Twenty-five microliters of diluted enzyme are added per well of a 96-well microfluor plate. The enzyme is then diluted in a 1: 4 ratio in the assay by the addition of inhibitor and substrate, obtaining a final concentration in the assay of 100mg / ml. mM stock solutions of inhibitors are prepared in dimethyl sulfoxide and then diluted in assay buffer as for the inhibitor dilution scheme, for the inhibition of human collagenase (MMP-1): Twenty-five microliters of each of the concentrations is added in triplicate to the microfluor plate. The final concentrations in the assay are 30 μM, 3 μM, 0.3 μM and 0.03 μM. The substrate (Dnp-Pro-Cha-Gly-Cys (Me) -His-Ala-Lys (NMA) -NH2) is prepared as for the inhibition of human collagenase (MMP-1) and 50 ml is added to each well. to give a final assay concentration of 10 μM. Fluorescence readings (excitation wavelength 360 nm, emission 450 nm) are made at time 0 and every 5 minutes, for 1 hour. Positive controls consist of enzyme and substrate without inhibitor and targets consist of substrate only. The IC5o are determined as for the inhibition of human collagenase (MMP-1). If it turns out that the IC50's are less than 0.03 μM, the inhibitors are then assayed at final concentrations of 0.3 μM, 0.03 μM, 0.003 μM and 0.0003 μM.

MMP-13 assay on collagen film Rat type-1 collagen was radiolabelled with C14 acetic anhydride (TE Cawston and AJ Barret, Anal. Biochem: 99, 340-345 (1979)) and used to prepare plates of 96 wells containing radiolabelled collagen films (Barbara Johson-Wint, Anal.Bíochem, 104, 175-181 (1980)). When a solution containing collagenase is added to the well, the enzyme cleaves the insoluble collagen that is unwound and, therefore, is solubilized. The activity of the collagenase is directly proportional to the amount of solubilized collagen, determined by the proportion of radioactivity released in the supernatant measured in a standard scintillation counter. The collagenase inhibitors are, therefore, compounds that reduce the radioactive counts released with respect to the controls without any inhibitor being present. A specific modality of this test is described in detail below. To determine the selectivity of compounds for MMP-13 against MMP-1 using collagen as a substrate, the following procedure is employed. proMMP-13 or proMMP-1, human, recombinant, is activated according to the previously described operative procedures. MMP-13 or activated MMP-1 is diluted to 0.6 ug / ml with buffer (50 mM Tris, pH 7.5, 150 mM NaCl, 10 mM Cacl2, 1 uM ZnCl2, 0.05% Brij-35, azide sodium 0.02%). Reserve (stock) solutions of test compound (10 mM) are prepared in dimethyl sulfoxide. Dilutions of the test compounds are made in the Tris buffer, above, at 0.2, 2.0, 20, 200, 2000 and 20,000 nM. 100 μl of the appropriate dilution of the drug and 100 μl of diluted enzyme are pipetted into the wells of a 96-well plate containing collagen-4C labeled collagen films. The final enzyme concentration is 0.3 μg / ml while the final drug concentration is 0.1, 1.0, 10, 100, 1,000 nM. Each of the drug and control concentrations is analyzed in triplicate. Controls are also made in triplicate for cases in which no enzyme is present and for enzyme in the absence of any compound. The plates are incubated at 37 ° C for a period of time such that about 30-50% of the available collagen is solubilized-determined by counting additional control wells at various time points. In most cases, about 9 hours of incubation are required. When the assay has progressed sufficiently, the supernatant from each well is separated and counted in a scintillation counter. The background counts (determined by the counts in the wells without enzyme) are subtracted from each sample and the% release is calculated with respect to the wells with enzyme alone and without inhibitor. The triplicate values of each point are averaged and the results are recorded on a graph as percent released versus drug concentration. The IC 50 are determined starting from the point where 50% inhibition of the release of the radiolabelled collagen is obtained. To determine the identity of the active collagenases in conditioned medium with cartilage, tests were carried out using collagen as a substrate, medium conditioned with cartilage containing collagenase activity and inhibitors of variable selectivity. The medium conditioned with cartilage was collected during the time in which the degradation of collagen was occurring and therefore, is representative of the collagenases responsible for the breakdown of collagen. The assays were carried out as explained above, except that instead of using recombinant MMP-13 or recombinant MMP-1, the source of the enzyme was conditioned with cartilage.

Degradation of cartilage collagen induced by IL-1, from bovine nasal cartilage This test uses explants of bovine nasal cartilage that are commonly used to test the efficiency of various compounds to inhibit or the degradation of proteoglycans induced by IL-1, or collagen degradation induced by IL-1. Bovine nasal cartilage is a tissue very similar to articular cartilage, that is, chondrocytes surrounded by a matrix that is mainly type II collagen and aggrecan. The tissue is used because: (1) it is very similar to articular cartilage, (2) it can be easily acquired, (3) it is relatively homogeneous, and (4) it is degraded with predictable kinetics after stimulation with IL-1. Two variations of this test have been used to test compounds. Both variations provide similar results. The two variations are described below: Variation 1 Three fragments of bovine nasal cartilage (2 mm diameter x 1.5 mm long, approximately) are placed in each of the wells of a 24-well tissue culture plate. One ml of medium without serum is then added to each well. Compounds are prepared in the form of 10 mM stock solutions in DMSO and then appropriately diluted in serum-free medium to the final concentrations, eg, 50, 500 and 5000 nM. Each of the concentrations is tested in triplicate. Recombinant human IL-1a (5 ng / ml) (IL-1) is added to control wells in triplicate and to each of the drug-containing wells. Control wells are also established in triplicate in which neither drug nor IL-1 have been added. The medium is separated and fresh medium containing IL-1 and the appropriate drug concentrations are added on days 6, 12, 18, and 24 or every 3-4 days if necessary. Separate media at each time point are stored at -20 ° C for later analysis. When the cartilage in the wells with only IL-1 has been reabsorbed almost completely (approximately on day 21), the experiment is terminated. The medium is separated and stored. Aliquots (100 μl) from each well at each time point are pooled, digested with papain and then analyzed for hydroxyproline content. The hydroxyproline obtained as background noise (average of the wells without IL-1 and without drug) is subtracted from each of the result points and the average is calculated for each triplicate. The results obtained in this way are expressed as a percentage of the mean value of IL-1 alone and are represented graphically. The IC50 is determined from this graph.

Variation 2 The experimental approach is the same as described above in variation 1, until day 12. On day 12, the conditioned medium is separated from each well and frozen. Then one ml of phosphate buffered saline (PBS) containing 0.5 μg / ml trypsin is added to each well, and the incubation is continued for a further 48 hours at 37 ° C. After 48 hours of incubation in trypsin, the PBS solution is separated. Aliquots (50 μl) of the PBS / trypsin solution and the two previous time points (days 6 and 12) are combined, subjected to hydrolysis and the hydroxyproline content is determined. The background hydroxyproline (average of the wells without IL-1 and without drug) is subtracted from each result point and the mean value is calculated for each triplicate. The result is then expressed as the percent of the average value IL-1 alone and is represented graphically. The IC50 is determined from this graph. In this variation, the time course of the experiment is shortened considerably. The addition of trypsin for 48 hours after 12 days of stimulation with IL-1 probably releases the type II collagen that has been damaged by collagenase activity but has not yet been released from the cartilage matrix. In the absence of stimulation with IL-1, treatment with trypsin produces only low background levels of collagen degradation in cartilage explants.

Inhibition of 92 kD qelatinase. Human (MMP-9) 92 kD gelatinase inhibition (MMP-9) is tested using the substrate Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 (10 μm) under similar conditions as above described for the inhibition of human collagenase (MMP-1). Human recombinant 92 kD gelatinase (MMP-9, gelatinase B) is activated for 2 hours with 1 mM p-aminophenylmercuric acetate (from a freshly prepared 100 mM stock solution in 0.2 N NaOH) at 37 ° C . 10 mM stock solutions in dimethyl sulfoxide, of inhibitors, are serially diluted in assay buffer (50 mM TRIS, pH 7.5, 200 mM NaCl, 5 mM CaCl 2, 20 μM ZnCl 2, 0.02% BRIJ-35 (vol. / vol.)) using the following scheme: 10 mM? 120 μM? 12 μM? 1.2 μM? 0.12 μM Other dilutions are made, as needed, following this same scheme. A minimum of four concentrations of inhibitor is carried out in each test to give one of the compounds. Then, 25 μl of each concentration is added to triplicate wells of a microfluor plate, 96-well U-bottom, black. Since the final assay volume is 100 μl, the final inhibitor concentrations are the result of another 1: 4 dilution (ie 30 μM, 3 μM, 0.3 μM, 0.03 μM, etc.). They are also prepared in triplicate, a blank (without enzyme or inhibitor) and an enzyme positive control (with enzyme and without inhibitor). Activated enzyme is diluted to 100 ng / ml in assay buffer, 25 μl per well is added to appropriate wells of the microplate. The final enzyme concentration in the assay is 25 ng / ml (0.27 nM). A 5 mM stock solution in substrate dimethyl sulfoxide (Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2) is diluted in assay buffer at 20 μM. The assay is initiated by the addition of 50 μl of diluted substrate obtaining a final assay concentration of 10 μM substrate. At time 0 a fluorescence reading is taken immediately (excitation wavelength 320 nm, emission 390 nm) and then readings are made every fifteen minutes, at room temperature, with a CytoFluor well plate reader from PerSeptive Biosystems with the gain in 90 units. The mean values of the fluorescence of the enzyme and the target are plotted against time. A preliminary time point is chosen on the linear part of this curve for IC50 determinations. Time point 0 for each of the compounds in each dilution is subtracted from the subsequent time point and the results are then expressed as both percent of enzyme control (fluorescence of the inhibitor divided by the fluorescence of positive control with enzyme x 100 ). The results are plotted as concentration of inhibitor against percentage of enzyme control. IC50's are defined as the concentration of inhibitor that gives a signal that is 50% of that of the enzyme positive control.

Assay of aqrecanase Primary porcine chondrocytes from articular joint cartilage are isolated by sequential digestion with trypsin and collagenase followed by digestion with collagenase overnight, and plated at 2 x 10 5 cells per well in 48-well plates with 5 μCi / ml of 35S (1000 Ci / mmoles) sulfur in plates coated with type I collagen. The cells are allowed to incorporate the marker in their proteoglycan matrix (approximately 1 week) at 37 ° C, under a 50% CO2 atmosphere . The night before the start of the assay, chondrocyte monolayers are washed twice with DMEM / 1% PSF / G and then allowed to incubate in fresh DMEM / 1% FBS overnight. The next morning the chondrocytes are washed once with DMEM / 1% PSF / G. The final wash is allowed to settle on the plates in the incubator while the dilutions are made. Media and dilutions can be prepared as described in the table below.

The plates are marked and only the 24 inner wells of the plate are used. In one of the plates, several columns are designated as IL-1 (without drug) and Control (without IL-1 or drug). These control columns are periodically counted to verify the release of 35S-proteoglycan. Control and IL-1 media are added to the wells (450 μl) followed by compound (50 μl) to start the assay in this way. The plates are incubated at 37 ° C with a 5% C02 atmosphere. At a release of 40-50% (when the CPM from the IL-1 media is 4-5 times that of the control media) determined by liquid scintillation counting (LSC) of media samples, the assay is given finished (9-12 hours). The media are separated from all wells and placed in scintillation tubes. Scintillation compound (scintillate) is added and radioactive beads (LSC) are acquired. To solubilize the cell layers, 500 μl of papain digestion buffer (0.2 M Tris, pH 7.0, 0.5 mM EDTA, 5 mM DTT and papain, 1 mg / ml) is added to each well. The plates with the digestion solution are incubated at 60 ° C during the hour. The cell layer is separated from the plates the next day and placed in scintillation tubes. Then the scintillation compound is added and the samples are committed to counting (LSC). The percentage of accounts released from the total present in each well is determined. The average values of the triplicates are obtained by subtracting from each well the background noise of each witness. The percent inhibition of the compound is based on samples of IL-1 and 0% inhibition (100% counts in total). The compounds of the present invention that were tested had an IC 50 of less than 1 μM, preferably less than 50 nM, in at least one of the tests described above. The compounds of the present invention also possess a different activity (ie, they are selective) for one or more reprolysins or MMPs. Selectivity, as used herein, alludes to the ratio of IC50 inhibitory results from two or more of the above protocols. The compounds of the invention that are selective have a ratio of at least 10. The compounds of the invention having the desired potency or selectivity can be identified by assaying a compound (preferably a small molecule, more preferably a hydroxamic acid and, most preferably, a compound of formula I) according to the protocols described above and determining the IC5o and the selectivity ratios. A group of preferred compounds (more preferably compounds of formula I) that can be identified by the methods of the present invention, include those inhibitors that possess selective activity against MMP-13 over MMP-1, (preferably a lower IC5o at 500 nM, more preferably 100 nM and, most preferably, 50 nM) for MMP-13 with a selectivity of at least 10 times, preferably 40 times, higher for MMP-13 than for MMP-1.

Preparation of compounds The following examples illustrate the preparation of the compounds of the present invention. The melting points are uncorrected. The NMR data are reported in parts per million (d). Commercial reagents were used without further purification. Chromatography refers to column chromatography carried out using 32-63 mm silica gel, performed under nitrogen pressure conditions (flash chromatography). The ambient temperature refers to 20-25 ° C. All non-aqueous reactions were performed under nitrogen atmosphere for convenience and to maximize yields.

EXAMPLE 1 Hydroxyamide of (4R) -1-r4- (4-fluorophenoxy) benzyl-1-oxo-midazolidine-4-carboxylic acid a) 1-benzyl ester, (4R) -3- [4- (4-fluorophenoxy) benzyl-1-oxo-imidazolidine-1,5-dicarboxylic acid 5-tert-butyl ester To a solution of the ester 1 - benzyl, 5-tert-butyl ester of (4R) -oxo-imidazolidine-5-dicarboxylic acid (650 mg, 2.0 mmol) in acetone (10 ml), powdered K2C03 (550 mg) and bromide of 4- ( 4-fluorophenoxy) benzyl (1.85 g, 6.6 mmol). The reaction mixture was stirred at room temperature for 6 days and then the solvent was evaporated. The residue was taken with ethyl acetate and washed with water and brine. After drying over MgSO 3, the solvent was evaporated. The title compound (820 mg, 78%) was isolated from the residue by chromatography on silica gel, eluting with chloroform. b) (4R) -1- [4- (4-Fluorophenoxy) benzyl-2-oxo-imidazolidine-4-carboxylic acid tert-butyl ester A solution of 1-benzyl ester, 5-tert-butyl acid ester ( 4R) -3- [4- (4-fluorophenoxy) benzyl] -2-oxo-imidazolidine-5-dicarboxylic acid (1.1 g, 2. 1 mmol) in methanol (100 ml), hydrogenated over 10% Pd on carbon (110 mg) at a pressure of 3 atmospheres, for 6 hours. After removing the catalyst by filtration through a nylon filter of 0.45 μm pore size, the solvent was evaporated to obtain the title compound (810 mg, 100%) as a yellow solid. c) (4R) -1-r4- (4-Fluorophenoxy) benzyl "| -2-oxo-midazolidine-4-carboxylic acid A solution of the (4R) -1- [4- (4 Fluorophenoxy) benzyl] -2-oxo-imidazolidine-4-carboxylic acid (180 mg, 1.56 mmol) in CH 2 Cl 2 (8 mL), treated with trifluoroacetic acid (8 mL) The reaction mixture was stirred at room temperature for 2.5 The concentration of the title compound, a white solid (297 mg, 58%) was collected by filtration, after triturating the oil with a mixture of diethyl ether and hexane warmed gently. d) (4R) -1- [4- (4-fluorophenoxy) benzin-2-oxo-imidazolidine-4-carboxylic acid (2-Tritymethylsilactyloxy) to a solution of the acid (4R) ) -1- [4- (4-fluorophenoxy) benzyl] -2-oxo-imidazolidine-4-carboxylic acid (120 mg, 0.36 mmol) in methylene chloride (5 ml), 1-hydroxybenzotriazole (73 mg, 0.54 mmole), diisopropylethylamine (0.13 ml, 0J5 mmole), O- (2-trimethylsilylethyl) hydroxylamine hydrochloride (92 mg, 0.54 mmole) and 1- [3- (dimethylamino) propyl] -3-ethylcarbodiimide hydrochloride (104 mg , 0.54 mmole). The reaction mixture was stirred at room temperature for 16 hours and then diluted with methylene chloride and water. The organic phase was washed successively with aqueous 1 M HCl solution, saturated aqueous NaHCO 3 solution and brine. After drying over MgSO4, the solution was concentrated to an oil. The title compound, an oil (85 mg, 53%) was isolated by chromatography on silica gel and elution with ethyl acetate. e) Hydroxyamide of (4R) -1-r4- (4-fluorophenoxy) benzyl-2-oxo-imidazolidine-4-carboxylic acid To a solution of (4R) -1- [(2-trimethylsilanylethoxy) -amide of [4R] 4- (4-fluorophenoxy) benzyl] -2-oxo-imidazolidine-4-carboxylic acid (85 mg, 0.19 mmol) in methylene chloride (5 ml), boron trifluoride etherate (0.073 ml, 0.58 mmol) was added. ). The reaction mixture was stirred at room temperature for 1.5 hours and then quenched by the addition of saturated aqueous solution of NH CI. The mixture was diluted with water and ethyl acetate and the organic phase was washed with brine, dried over MgSO4 and concentrated, obtaining a white solid. The title compound (31 mg, 47%) was isolated by recrystallization from a mixture of ethyl acetate and methanol. 1 H NMR (DMSO-de): d 10.63 (broad s, 1 H), 8.93 (s, broad, 1 H), 7.23-7.17 (m, 4H), 7.05-7.01 (m, 2H), 6.92 (d, J = 8J Hz, 2H), 6J6 (s, 1 H), 4.22 (d, J = 15.2 Hz, 1 H), 4.15 (d, J = 15.2 Hz, 1 H), 3.92-3.89 (m, 1 H) ), 3.40 (apparent t, J = 9.1 Hz, 1 H), 3.15-3.12 (m, 1 H). MS m / z 344 (m-1). Analysis calculated for C? 9H? 6FN304: C, 59.13; H, 4.67; N, 12.17. Found: C, 58.98; H, 4.83; N, 12.10.

EXAMPLE 2 (4R) -1-f4-naphthalen-1-yloxy) benzin-2-oxo-imidazolidine-4-carboxylic acid hydroxyamide MS m / z 376 (M-1). Analysis calculated for C21H? 9N304: C, 66.83; H, 5.07; N, 11.13. Found: C, 66.75; H, 5.30; N, 11.13.

EXAMPLE 3 (4R) -1-r4- (naphthalen-2-yloxy) benzyl-2-oxo-imidazolidine-4-carboxylic acid hydroxyamide MS m / z 376 (M-1). Analysis calculated for C21H19N304 + 0.5 H20: C, 65.28; H, 5.22; 10.87. Found: C, 65.01; H, 5.12; N, 11.28.

EXAMPLE 4 (4R) -1- (4-methoxybenzyl) -2-oxo-midazolidine-4-carboxylic acid hydroxyamide p.f. 130-133 ° C. MS m / z 264 (M-1).

Analysis calculated for C 12 H 15 N 304: C, 54.33; H, 5.70; N, 15.84. Found: C, 54.24; H, 51.77; N, 15.62.

EXAMPLE 5 Hydroxyamide of (4R) -1-r3- (4-fluorophenoxy) benzyl-2-oxo-imidazolidine-4-carboxylic acid MS m / z (M-1). Analysis calculated for C19H? 6FN304: C, 59.13; H, 4.67; N, 12.17. Found: C, 59.24; H, 4.60; N, 12.42.

EXAMPLE 6 (4R) -1-Naphthalen-2-ylmethyl-2-oxo-imidazolidine-4-carboxylic acid hydroxyamide MS m / z 284 (M-1). Analysis calculated for C? 5H15N303: C, 63.15; H, 5.30; N, 14.73. Found: C, 62.82; H, 5.32; N, 14.49.

EXAMPLE 7 (4R) -1- (4'-Fluorobiphenyl-4-ylmethyl) -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide MS m / z 328 (M-1). Analysis calculated for C 7 H 6 FN 3 O 3 + 0.5 H 20: C, 60.35; H, 5.06; N, 12.42. Found: C, 60.43; H, 4.99; N, 12.83.

EXAMPLE 8 (4R) -1 - (4-Benzyloxybenzyl) -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide MS m / z 340 (M-1). Analysis calculated for C? 8H? 9N304: C, 63.33; H, 5.61; N, 12.31. Found: C, 63.13; H, 5.62; N, 12.28.

EXAMPLE 9 (4R) -1-r4- (2-Chloro-4-fluorobenzyloxy) benzyl-2-oxo-imidazolidine-4-carboxylic acid hydroxyamide MS m / z 392, 394 (M-1).

Analysis calculated for C 8H17CIFN3O4 + 0.5 H20: C, 53.67; H,. Found: C, 53.78; H, 4.51; N, 10.15.

Claims (19)

NOVELTY OF THE INVENTION CLAIMS

1. - A compound according to formula (I) or a pharmaceutically acceptable salt or solvate thereof, wherein R1 is C6-C10 aryl, Ci-Cg heteroaryl, aryl (C6-C? o) C1-C6 alkyl, heteroaryl (C? -Cg) alkyl, C? -C6, aryl (C6-C? O) aryl of C6-C? 0, heteroaryl (CrCg) aryl of C6-Cio, aryl (C6-C? O) heteroaryl of C1-C9, heteroaryl (CrCg) heteroaryl of C1-C9, aryl (C6-C? o) oxyaryl of C6-C? 0, heteroaryl (C? -Cg) oxoaryl of C6-C? 0, aryl (C6-C? o) C 1 -C 9 oxyheteroaryl, C 1 -C 9 heteroaryl (CrCg) oxyheteroaryl, C 1 -C 6 aryl (C 6 -C 7) oxyalkyl, C 6 -C 6 heteroaryl (C 1 -C 9) oxyalkyl, aryl (C 6) C? O) alkyl (C? -C6) aryl of C6-C? 0, heteroaryl (C? -Cg) alkyl (C? -Ce) aryl of Ce-Cio, aryl (C6-C1o) alkyl (C1-Ce) ) C Cg heteroaryl, Ci-CgJalqui heterocyclic C6 heteroaryl Cg C heteroaryl, C6-C6 aryl (C6-C0o) C6-C6 alkoxy, heteroaryl (Cr Cg) alkoxy (C6-C6) ) aryl of Ce-Cio, aryl (C6-C? o) alkoxy (C? -C6) heteroaryl of Ci-Cg, heteroaryl (C? -C9) alkoxy (C Ce) heteroaryl of C Cg, aryl (C6-C? 0) oxyalkyl (d-C6) C6-C? Aryl, heteroaryl (Ci-Cg) oxyalkyl- (C? -C6) a Rile of C6-C10, aryl (C6-Cio) oxyalkyl (C? -C6) heteroaryl of Ci-Cg, heteroaryl (Ci-Cg) oxyalkyl (d-C) heteroaryl of Ci-Cg, aryl (C6-d0) aryl (C6-C? 0) d-C6 alkyl, heteroaryl (d-C9) -aryl (C6-C10) alkyl of CrC6, aryl (C6-C? 0) heteroaryl (Ci-Cg) alkyl of Cr C6, heteroaryl (Ci-Cg) heteroaryl (Ci-Cg) C-Cß alkyl, (C6-C?) Aryl (C?-C6) alkoxy C-C6 alkyl, or heteroaryl (CrCg) (C 1 -C 6) alkoxy C Cß, wherein, independently, each of the ring carbon atoms of said Ce-Cι aryl moieties and C?-C 9 heteroaryl which is capable of forming an additional bond, is optionally substituted with a selected group between fluoro, chloro, bromo, C? -C6alkyl, d-C6alkoxy, and perfluoroalkyl of C C3 and perfluoroalkoxy of C C3; R2 and R3 are each independently selected from hydrogen and C6-C6 alkyl, or taken together they form a spiro-compound ring of the formula wherein X is a bond, CH2, O, S, NH or N-Ci-Cß alkyl, n is independently 1 or 2, and m is independently 1 or 2; and R 4 is hydrogen or CrC 6 alkyl.

2. The compound according to claim 1, of the formula (I ') (?)

3. The compound according to claim 1, wherein R2, R3 and R4 are hydrogen.

4. The compound according to claim 1, wherein R4 is hydrogen.

5. The compound according to claim 1, wherein R4 is d-C6 alkyl.

6. The compound according to claim 1, wherein R2 and R3 are hydrogen.

7. The compound according to claim 1, wherein R2 and R3 are C6 alkyl, so that R2 is the same as R3.

8. The compound according to claim 1, wherein R2 and R3 taken together form a spirocomposite ring of the formula wherein X is a bond, CH2, O, S, NH or N-C-C-alkyl, n is 1 or 2, and m is 1 or 2, so that n is the same as m.

9. The compound according to claim 1, wherein R1 is 4- (4-fluorophenoxy) phenyl.

10. The compound according to claim 1, wherein R1 is 4- (4-chlorophenoxy) phenyl.

11. The compound according to claim 1, wherein R1 is 4- (naphthalen-2-yloxy) phenyl.

12. The compound according to claim 1, selected from the group consisting of: (4R) -1- [4- (4-fluorophenoxy) benzyl] -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4R) -1 - [4-naphthalen-1-yloxy) benzyl] -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4R) -1- [4-naphthalen-2-yloxy) benzyl] -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4f?) - 1- (4-methoxybenzyl) -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4R) -1 - [3- (4-fluorophenoxy) benzyl] -2-oxo-imidazole-4-carboxylic acid hydroxyamide; (4) -1-naphthalen-2-ylmethyl-2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4R) -1 - (4'-Fluorobiphenyl-4-ylmethyl) -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4R) -1- (4-benzyloxybenzyl) -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; and (4R) -1- [4- (2-Chloro-4-fluorobenzyloxy) benzyl] -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide.

13. The compound according to claim 1, selected from the group consisting of: (4 /?) - 1- [4- (4-chlorophenoxy) benzyl] -2-oxo-imidazolidine-4-hydroxyamide carboxylic; hydroxyamide of (4rR) -1- [4- (4-fluorophenoxy) benzyl] -2-oxo-7-oxa-! 3-diazaspiro [4,4] nonane-4-carboxylic acid; hydroxyamide of (4r?) - 2-oxo-1- [4- (pyridin-4-yloxy) benzyl] imidazolidine-4-carboxylic acid; (4R) -4-methyl-2-oxo-1 - [4- (pyridin-4-yloxy) benzyl] imidazolidine-4-carboxylic acid hydroxyamide; (4) -5,5-Dimethyl-2-oxo-1- [4- (pyridin-4-yloxy) -benzyl] -amidazolidine-4-carboxylic acid hydroxyamide; (4f?) - 1- [4- (4-fluorophenoxy) benzyl] -5,5-dimethyl-2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4f?) - 1- [4- (4-chlorophenoxy) benzyl] -5,5-dimethyl-2-oxo-midazolidine-4-carboxylic acid hydroxyamide; (4R) -1 - [4- (4-fluorophenoxy) benzyl] -4-methyl-2-oxo-imidazole-4-carboxylic acid hydroxyamide; (4R) -1- [4- (4-chlorophenoxy) benzyl] -4-methyl-2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; hydroxyamide of (4) -1- [4- (4-fluorophenoxy) -benzyl] -2-oxo-1,3-diazaspiro [4,4] nonane-4-carboxylic acid hydroxyamide; (4R) -1- [4- (4-chlorophenoxy) benzyl] -2-oxo-1,3-diazaspiro [4,4] nonane-4-carboxylic acid hydroxyamide; (4) -1- [4- (4-fluorophenoxy) benzyl] -4,5,5-trimethyl-2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4R) -1- [4- (4-chlorophenoxy) benzyl] -4,5,5-trimethyl-2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4) -4-methyl-1- [4- (naphthalen-2-ylox!) benzyl] -2-oxo-midazolidine-4-carboxylic acid hydroxyamide; (4R) -5,5-dimethyl-1- [4- (naphthalen-2-yloxy) benzyl] -2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; (4?) - 1- [4- (5-fluoropyridin-2-yloxy) benzyl] -4-methyl-2-oxo-imidazolidine-4-carboxylic acid hydroxyamide; hydroxyamide of (4R) -2-oxo-1- (4-pyridin-4-ylbenzyl) imidazolidine-4-carboxylic acid, and (4) -2-oxo-1- (4-pyridylmethyl) imidazolidine hydroxyamide -4-carboxylic acid.

14. A pharmaceutical composition for the treatment of a state selected from the group consisting of arthritis (including osteoarthritis and rheumatoid arthritis), inflammatory bowel disease, Crohn's disease, emphysema, chronic obstructive pulmonary disease, Alzheimer's disease, toxicity of organ transplantation, cachexia, allergic reactions, inflammation, allergic contact hypersensitivity, cancer, tissue ulceration, restenosis, periodontal disease, epidermolysis bullosa, osteoporosis, loosening of implants of artificial joints, atherosclerosis (including rupture of atherosclerotic plaques) aneurysm aortic (including abdominal aortic aneurysm and cerebral aortic aneurysm), congestive heart failure, myocardial infarction, stroke, cerebral ischemia, head trauma, spinal cord damage, neurodegenerative disorders (acute and chronic), autoimmune disorders, disease of Huntington, Parkinson's disease, headaches, depressions, peripheral neuropathies, pain, cerebral amyloid angiopathy, nootropic or knowledge improvement, amyotrophic lateral sclerosis, multiple sclerosis, ocular angiogenesis, corneal damage, macular degeneration, abnormal healing of wounds, burns, diabetes, invasion tumor, tumor growth, tumor metastasis, corneal scars, scleritis, AIDS and sepsis and septic shock in a mammal, including the human being, comprising an amount of a compound of formula I effective in such treatment, and a pharmaceutically acceptable excipient.

15. The use of a compound as claimed in claim 1, for the manufacture of a medicament for treating a condition selected from the group consisting of arthritis (including osteoarthritis and rheumatoid arthritis), inflammatory bowel disease, Crohn's disease, emphysema, chronic obstructive pulmonary disease, Alzheimer's disease, organ transplant toxicity, cachexia, allergic reactions, inflammation, allergic hypersensitivity by contact, cancer, tissue ulceration, restenosis, periodontal disease, epidermolysis bullosa, osteoporosis, loosening of implants Artificial joints, atherosclerosis (including rupture of atherosclerotic plaques), aortic aneurysm (including abdominal aortic aneurysm and cerebral aortic aneurysm), congestive heart failure, myocardial infarction, 5 strokes, cerebral ischemia, head trauma, spinal cord damage , neuro-degenerative disorders (acute and chronic), autoimmune disorders, Huntington's disease, Parkinson's disease, «*» Migraines, depressions, peripheral neuropathies, pain, cerebral amyloid angiopathy, nootropic or knowledge improvement, amyotrophic lateral sclerosis, 10 multiple sclerosis, ocular angiogenesis, corneal damage, macular degeneration, abnormal wound healing, burns, diabetes, tumor invasion, tumor growth, tumor metastasis, corneal scars, scleritis, AIDS and sepsis and septic shock in a mammal, including the human being.

16.- A pharmaceutical composition for the treatment of a A condition that can be treated by inhibiting matrix metalloproteinases in a mammal, including a human, comprising an amount of a compound according to claim 1, effective in such treatment, and a pharmaceutically acceptable excipient.

17. A pharmaceutical composition for the treatment of a A condition that can be treated by inhibiting a mammalian reprolysin in a mammal, including a human, comprising an amount of a compound according to claim 1, effective in such treatment and a pharmaceutically acceptable excipient.

18. The use of a compound as claimed in claim 1, for the manufacture of a medicament for the inhibition of matrix metalloproteinases in a mammal, including a human.

19. The use of a compound as claimed in claim 1, for the manufacture of a medicament for inhibiting a mammalian reprolysin in a mammal, including a human. ar- The present invention relates to a compound of the formula wherein R1, R2, R3 and R4 are as defined above and their pharmaceutically acceptable salts and solvates, which are useful, for example, as inhibitors of matrix metalloproteinases; the present invention is also directed to pharmaceutical compositions comprising such compounds and to methods of treating diseases such as osteoarthritis, rheumatoid arthritis, cancer, osteoporosis, tissue ulceration, restinosis, periodontal disease, inflammation, epidermolysis bullosa, scleritis, stroke, disease of Alzheimer's, and the like, characterized by inappropriate activity of matrix metalloproteinases; Methods of synthesis of the compounds of formula (I) are also described. PFIZER / pbg * yac * yrc * tpr * kra * igp * mmf * jtc * cgm * aom / jtc