Amides, phosphinic acid and a method of prevention or treatment of a disease associated with unwanted activity metalloprotease

 

(57) Abstract:

The invention relates to Amida phosphinic acids f-crystals (I), where R1is hydrogen, alkyl, phenylalkyl, pyridinyl, pyridinylmethyl, alkoxyalkyl, generalkonsulat; R2is hydrogen, alkyl, phenylalkyl, indolyl, generalkonsulat, alkylthiomethyl, acylaminoalkyl; R3- alkyl or phenyl; R4- alkyl, phenyl or substituted phenyl, pyridyl, thienyl or furyl, to their optical isomers, diastereomers, enantiomers, pharmaceutically acceptable salts or biohydrology esters that can be used as inhibitors of matrix metalloprotease in the treatment of conditions characterized by excessive activity of these enzymes. 3 S. and 7 C.p. f-crystals, 2 PL.

The invention relates to compounds useful for the treatment of diseases related to activity of metalloprotease, in particular the activity of zinc metalloprotease.

A number of structurally related metalloprotease [MPs, MP] structural breaks down proteins. These metalloprotease often affect intercellular material and, therefore, participate in the destruction and remodeling of tissue. Such proteins are called metalloprotease or MP. ), The P and their examples are already known.

These include MP matrix metalloproteinase [MMP], zinc metalloprotease, many of metalloprotease associated with membranes, TNF-converting enzyme, angiotensin-converting enzymes (ACEs, ACE), decomposers, including ADAMs (see Wolfsberg et al, 131 J. Cell Bic. 275-78, October 1995), and enkephalinase. Examples of MP include fibroblast collagenase human skin, gelatinase of fibroblast of the human skin collagenase, arakanese and gelatinase sputum person and stromelysin person. It is believed that the collagenase, stromelysin, arakanese and related enzymes important in mediating the symptomatology of many diseases.

Possible therapeutic indications inhibitors MP already described in the literature. See, for example, U.S. patent N 5506242 (Ciba Geigy Corp.); U.S. patent N 5403952 (Merck & Co.); PCT published application WO 96/06074 (British Bio Tech Ltd); PCT publication WO 96/00214 (Ciba Geigy); WO 95/35275 (British Bio Tech Ltd); WO 95/35276 (British Bio Tech Ltd); WO 95/33731 (Hoffman-LaRoche); WO 95/33709 (Hoffman-LaRoche); WO 95/32944 (British Bio Tech Ltd); WO 95/26989 (Merck); WO 95/29892 (DuPont Merck); WO 95/24921 (Inst. Ophthalmology); WO 95/23790 (Smith Kline Beecham); WO 95/22966 (Sanofi Winthrop); WO 95/19965 (Glycomed); WO 95/19956 (British Bio Tech Ltd); WO 95/19957 (British Bio Tech Ltd); WO 95/19961 (British Bio Tech Ltd); WO 95/13289 (Chiroscience Ltd.); WO 95/12603 (Syntex); WO 95/09633 (Florida State Univ. ); WO 95/09620 (Florida State Univ.); WO 95/04033 (Celltech); WO 94/25434 (Celltech); WO 94/25435 (Celltech); WO 93/14112 (Mer82598 (Merck) and GB 2268934 (British Bio Tech Ltd); published applications, European patent EP 95/684240 (Hoffman LaRoche); EP 574758 (Hoffman LaRoche); EP 575874 (Hoffman LaRoche); published patent application Japan JP 08053403 (Fujusowa Pharm. Co. Ltd); JP 7304770 (Kanebo Ltd. ); and Bird et al, J. Med. Chem. volume 37, pp. 158-69 (1994). Examples of possible therapeutic applications of MP inhibitors include rheumatoid arthritis (Mullins D. E., et al., Biochim. Biophys. Acta (1983) 695: 117-214); osteoarthritis (Henderson, B. , et al. Drugs of the Future (1990) 15: 495-508); metastasis of tumor cells (ibid, Broadhurst, M. J., application N 276436 for the European patent (published in 1987), Rech, R., et al., 48 Cancer Res. 3307-3312 (1988)); and various ulcers or ulcerative condition of the tissue. For example, ulcerative conditions can arise in the cornea as a result of burns from alkali or as a result of infection by the bacteria Pseudomonas aeruginosa, Acanthamoeba spp., herpes viruses simple and vaccinii.

Other examples of conditions characterized by undesired activity of metalloprotease include periodontal disease, congenital bullous bullosa, fever, inflammation and scleritis (cf. DeCicco et al., WO 95/29892, published on 9 November 1995).

Due to the involvement of such metalloprotease in the creation of many painful conditions, attempts were made to obtain inhibitors of these enzymes. A number of these inhibitors are described in liters 1991 in the name of Handa et. al.; U.S. patent N 4771038, issued September 13, 1988 in the name of Wolanin, et al.; U.S. patent N 4743587, issued may 10, 1988 in the name of Dickens, et al.; European patent publication N 575844, published on December 29, 1993 by Broadhurst, et al., international patent publication N WO 93/09090, published may 13, 1993 by Isomura et al.; international patent publication 92/17460, published October 15, 1992 by Markwell et al.; and European patent publication N 498665, published August 12, 1992 by Beckett et al.

As a method of treatment of diseases associated with unwanted activity metalloprotease, it would be advisable inhibition of these metalloprotease. Although it has received a lot of different inhibitors, however, continues to be a need for powerful inhibitors of the matrix metalloprotease suitable for treatment of such diseases.

The aim of the present invention is to provide potent inhibitors of metalloprotease.

Another objective of the present invention is to provide pharmaceutical compositions containing such inhibitors.

The next objective of the present invention is to provide a method for treatment of diseases associated with metalloprotease.

In accordance with this is effective in the treatment of conditions characterized by excessive activity of these enzymes. In particular, the present invention relates to a compound having a structure corresponding to formula (I)

< / BR>
where R1i represents hydrogen, alkyl, arylalkyl, heteroseksualci, alkoxyalkyl, allakariallak or alkylthiomethyl;

R2represents hydrogen, alkyl, arylalkyl, heteroseksualci, alkoxyalkyl, allakariallak or alkylthiomethyl;

R3represents alkyl, cycloalkyl, carbocyclic or heterocyclic aryl, hydroxyalkyl, alkoxyalkyl or aminoalkyl; and

R4represents carbocyclic or heterocyclic aryl;

its optical isomer, diastereoisomer, or enantiomer, or its pharmaceutically acceptable salt or biogerontology alkoxyamino, complex ether, allacciamento or imide.

Preferred R4includes phenyl and substituted phenyl. The preferred substitution at R4produced near the place of attachment or directly opposite (i.e., if R4represents phenyl, in positions 2 and/or 4). Preferred phenyl substituents include halogen, alkyl, alkoxy, nitro, cyano and the like. Preferred R3is alteredactions - H or C1-C4alkyl. Preferred R1is H or alkyl, arylalkyl, and more preferred is C1-C6alkyl or aryl(C1-C2) alkyl.

These compounds possess the ability to inhibit at least one matrix metalloprotease mammal. So in other aspects the present invention relates to pharmaceutical compositions containing the compounds of formula (I), and methods of treating diseases characterized by the activity of the matrix of metalloprotease, using these compounds or pharmaceutical compositions containing these compounds.

Matrix metalloprotease, especially active in the wrong place (for example, the body or some types of cells), can be done by the target by coupling compounds of the present invention with a targeting ligand specific to the token in this place, such as an antibody or its fragment, or a receptor ligand. Methods of conjugation are known in this field of chemistry.

The present invention also concerns many other ways to take advantage of the unique properties of these compounds. Thus, in another aspect of the present izobret as having affinity reagents for purification of the desired matrix metalloprotease.

In one aspect, the present invention relates to compounds of formula (I) with the entered label. When the compounds of the present invention are associated with at least one matrix by metalloproteases, the label can be used to detect a relatively high level matrix metalloprotease in vivo or cell culture in vitro.

In addition, the compounds of formula (I) can be associated with the media, allowing the use of these compounds in the schemes of immunization to generate antibodies specifically immunoreactive with the compounds of the present invention. Typical methods of conjugation are known in this field. These antibodies, in this case, useful for therapy, and to control the dosing of inhibitors.

Compounds of the present invention are inhibitors of the matrix metalloprotease mammals. The compounds are preferably a compound of formula (I) or its pharmaceutically acceptable salt, or biohydrology alkoxyamino, alloxanic or imide.

Used terms and definitions

"Acyl" or "carbonyl" is a radical, which can be formed by removing the hydroxy-group of the carboxylic

"Acyloxy" is oxely radical having the acyl substituent (i.e.,- O - acyl), for example,- O-C(=O)-alkyl.

"Alkoxyaryl" is the acyl radical (-C(=O)-), with CNS Deputy (i.e.,- O-R), for example-C(=O)-O-alkyl. This radical can be called complex ether.

"Acylamino" represents an amino radical having the acyl substituent (i.e.,- N-acyl), for example-NH-C(=O)-alkyl.

"Alkenyl" represents an unsubstituted or substituted hydrocarbon radical having 2 to 15, preferably 2-10, and more preferably 2-8 carbon atoms, unless otherwise indicated. Alkeneamine deputies have at least one olefinic double bond (including, for example, vinyl, allyl and butenyl).

"Quinil" represents an unsubstituted or substituted hydrocarbon radical having 2 to 15, preferably 2-10, and more preferably 2-8 carbon atoms, unless otherwise indicated. The chain has at least one triple carbon-carbon bond.

"Alkoxy" is an oxygen radical having a hydrocarbon Deputy, where the hydrocarbon chain is an alkyl or alkenyl (i.e.,- O-alkyl or-O-alkenyl). Preferred alkoxygroup include, for example, meoment, substituted alkoxy fragment (i.e.,- alkyl-O-alkyl). It is preferred, when the alkyl has 1-6 carbon atoms (more preferably 1-3 carbon atoms and the alkoxy has 1 to 6 carbon atoms (more preferably 1-3 carbon atoms).

"Alkyl" represents an unsubstituted or substituted saturated hydrocarbon radical having 1-15, preferably 1-10 and more preferably 1-4 carbon atoms, unless otherwise indicated. Preferred alkyl groups include, for example, substituted or unsubstituted methyl, ethyl, propyl, isopropyl and butyl.

When referred to in this description the term "spirits" or "spiritlessly" refers to cyclic fragment, having total carbon atoms with another ring. Such cyclic fragment can be, by their nature, carbocyclic or heterocyclic. Preferred heteroatoms included in the basis of heterocyclic spirocyclic include oxygen, nitrogen and sulfur. Spirocyclic can be unsubstituted or substituted. Preferred substituents include oxo, hydroxy, alkyl, cycloalkyl, arylalkyl, alkoxy, amino, heteroalkyl, aryloxy condensed rings (e.g., bettiol, cycloalkyl, heterocyclyl what oatom heterocycle may be substituted, if you valency. The preferred size spirocycles rings include a 3-7-membered ring system.

Alkylene refers to alkyl, alkenyl or quinil, which is not radical, and diradical. Similarly "heteroalkyl" can be defined as (diradical)-alkylen having in its chain heteroatom.

"Alkylamino" is amino radical having one (secondary amine) or two (tertiary amine) alkyl substituent (i.e.,- N-alkyl); for example, methylamino (-NHCH3), dimethylamino (N(CH3)2), methylethylamine (-N(CH3)CH2CH3).

"Aminoacyl" represents an acyl radical, having an amino substituent (i.e.,- C(=O)-N); for example, -C(=O)-NH2. The amino aminoacylase fragment may be unsubstituted (i.e., primary amine) or substituted one (secondary amine) or two (i.e., tertiary amine) alkyl groups.

"Aryl" represents an aromatic carbocyclic ring radical. Preferred aryl groups include, for example, phenyl, tolyl, xylyl, cumenyl, naphthyl, diphenyl and fluorenyl. Such groups can be substituted or unsubstituted.

"Arylalkyl" represents alkali phenylpropyl. Such groups can be substituted or unsubstituted.

"Arylalkylamine" is amino radical, substituted aryl-alkyl group (for example, -NH-benzyl). Such groups can be substituted or unsubstituted.

"Arylamino" is amino radical, a substituted aryl group (e.g.,- NH-aryl). Such groups can be substituted or unsubstituted.

"Aryloxy" is an oxygen radical, aryl having a substituent (i.e.,- O-aryl). Such groups can be substituted or unsubstituted.

"Carbocyclic ring" is an unsubstituted or substituted, saturated, unsaturated or aromatic hydrocarbon ring radical. Carbocyclic rings are monocyclic or condensed with bridge or spironolactone ring systems. Monocyclic carbocyclic rings usually contain 4 to 9, preferably 4 to 7 atoms. Polycyclic carbocyclic rings contain 7-17, preferably 7-12 atoms. Preferred polycyclic system containing 4-, 5-, 6 - or 7-membered ring condensed with a 5-, 6 - or 7-membered rings.

"Carbocyclic" represents an unsubstituted or substituted acticle is aryl or cycloalkyl, more preferably the aryl. Preferred carbocycle-alkyl groups include benzyl, phenylethyl and phenylpropyl.

"Carbocyclization" is unsubstituted or substituted heteroalkyl radical, a substituted carbocyclic ring. Unless otherwise stated, carbocyclic ring, preferably an aryl or cycloalkyl, more preferably an aryl. Heteroalkyl preferably represents 2-oxopropyl, 2-oxoethyl, 2-thiopropyl or 2-deatil.

"Carboxylic" is unsubstituted or substituted alkyl radical, substituted carboxy (-C(=O)OH) fragment. For example, - CH2-C(=O)OH.

"Cycloalkyl" represents a saturated carbocyclic ring radical. Preferred cycloalkyl groups include, for example, cyclopropyl, cyclobutyl and cyclohexyl.

"Cyclogeranyl" represents a saturated heterocyclic ring. Preferred cyclogyro-alkyl groups include, for example, morpholinyl, piperidinyl, piperazinil, tetrahydrofuryl and hydantoinyl.

"Condensed ring" is a ring combined with each other so that they have two common ring atom. This ring can be scandens rouillie, aryl and heterocyclic radicals or the like.

"Heteroseksualci" is an alkyl radical, a substituted heterocyclic ring. Heterocyclic ring is preferably heteroaryl or cyclogeraniol, and more preferably heteroaryl. Preferred heteroseksualci includes C1-C4alkyl with an attached preferred heteroaryl. More preferred is, for example, pyridylethyl etc.

"Heterocyclisation" represents an unsubstituted or substituted heteroalkyl radical, a substituted heterocyclic ring. Heterocyclic ring is preferably an aryl or cyclogeraniol, and more preferably an aryl.

"Heteroatom is a nitrogen atom, sulfur or oxygen. Group having one or more heteroatoms, may contain different heteroatoms.

"Heteroalkyl" represents an unsubstituted or substituted unsaturated radical having 3 to 8 members containing carbon atoms and one or two heteroatoms. The chain has at least one double carbon-carbon bond.

"Heteroalkyl" represents an unsubstituted or Zam

"Heterocyclic ring is unsubstituted or substituted, saturated, unsaturated or aromatic ring radical which consists of carbon atoms and one or more heteroatoms in the ring. Heterocyclic rings are monocyclic or condensed with bridge or spironolactone ring systems. Monocyclic heterocyclic ring containing 3 to 9, preferably 4 to 7 atoms. Polycyclic rings contain 7-17, preferably 7-13 atoms.

"Heteroaryl" represents an aromatic heterocyclic ring or a monocyclic or bicyclic radical. Preferred heteroaryl groups include, for example, thienyl, furyl, pyrrolyl, pyridinyl, pyrazinyl, thiazolyl, pyrimidinyl, chinoline and tetrazolyl, benzothiazolyl, benzofuran, indolyl and the like. Such groups can be substituted or unsubstituted.

"Halo", "halogen" or "halide" is a radical as chlorine atom, bromine, fluorine or iodine. Preferred radicals are bromine, chlorine and fluorine.

In addition, when referred to in this specification, "lower" hydrocarbon fragment (for example, "lower alkyl) represents a hydrocarbon cataflam a cationic salt, formed at any acidic (e.g., carboxyl) group, or an anionic salt formed at any basic (e.g., amino) group. Many such salts are known in the art, for example those described in international patent publication 87/05297 (Johnston et al.), published September 11, 1987 (included in this description by reference). Preferred cationic salts include alkali metal salts (such as sodium and potassium), and salts of alkaline earth metals (such as magnesium and calcium) and organic salts. Preferred anionic salts include halides (such as chlorides).

"Biohydrology alkoxyamino" or "biohydrology allocine" - it amides, hydroxamic acids, which do not interfere with the inhibitory activity of compounds or readily converted in vivo human or lower animal in active hydroxamic acid.

"Biohydrology hydroxyamide" is the imide compounds of formula (I), which does not interfere with inhibitory activity of compounds against metalloprotease or easily converted in vivo human or lower animal in the active compound of the formula (I). Such hydroxyamide include those that do not interfere with the biological activity of the compounds of the AET inhibitory activity of these compounds against metalloprotease or easily converted in the animal body to the active compound of the formula (I).

"MES" is a complex formed by combining a solute (e.g., hydroxamic acid) and solvent (e.g. water). See J. Honig et al. The Van Nostrand Chemist''s Dictionary, page 650 (1953). Pharmaceutically acceptable solvents used in accordance with the present invention include those that do not interfere with the biological activity of hydroxamic acids (for example, water, ethanol, acetic acid, N,N-dimethylformamide and other known or easily determined by a qualified technician).

"Optical isomer", "stereoisomer", "diastereoisomer" when referred to in this description are well-known standard values (I.e., Hawley's Condensed Chemical Dictionary, 11th edition).

The illustration of a particular protected forms and other derivatives of the compounds of formula (I) should not be considered limiting. Other useful protective groups, types of salts, etc. is the responsibility of a qualified professional.

As mentioned above, when used in the present invention substituting groups may themselves be substituted. Such substitution may be made by one or more substituents. Such substituents include those perechisleny. Preferred substituents include, for example, alkyl, alkenyl, alkoxy, hydroxy, oxo, nitro, amino, aminoalkyl (for example, aminomethyl and so on ), cyano, halogen, carboxy, alkoxyalkyl (for example, carboethoxy and so on), thiol, aryl, cycloalkyl, heteroaryl, heteroseksualci (for example, piperidinyl, morpholinyl, pyrrolidinyl and so on), imino, thioxo, hydroxyalkyl, aryloxy, arylalkyl and combinations thereof.

When used in this description of "the matrix metalloprotease mammal" means a metal-containing enzyme found in mammals that can catalyze the cleavage of collagen, gelatin or of proteoglycan in suitable conditions analysis. Suitable conditions analysis can be found, for example, in U.S. patent N 4743587, where there was a reference to the method of Cawston, et al. Anal. Biochem. (1979) 99: 340-345, and the use of a synthetic substrate described by Weingarten, N., et al., Biochem. Biophy. Res. Comm. (1984) 139: 1184-1187. Of course, can be applied to any standard method of analysis of the splitting of these structural proteins. Everything mentioned in this description matrix metalloprotease are the zinc-containing proteases that are similar in structure to, for example, human stromelysin or fibroblastoid the collagenase in the skin. The way the above methods of analysis. To confirm the inhibitory activity of the compounds of the present invention can use a dedicated matrix metalloprotease or crude extracts, contains a number of enzymes capable of tissue destruction.

Connection

Compounds of the present invention described in "Summary of the invention". Preferred compounds of formula (I) include a connection

< / BR>
where R1represents hydrogen, alkyl, arylalkyl, heteroseksualci, alkoxyalkyl, allakariallak or alkylthiomethyl;

R2represents hydrogen, alkyl, arylalkyl, heteroseksualci, alkoxyalkyl, allakariallak or alkylthiomethyl;

R3represents alkyl, cycloalkyl, carbocyclic or heterocyclic aryl, hydroxyalkyl, alkoxyalkyl or aminoalkyl; and

R4represents carbocyclic or heterocyclic aryl;

its optical isomer, diastereoisomer, or enantiomer, or its pharmaceutically acceptable salt or biohydrology alkoxyamino, ester, alloxanic or imide.

Obtaining compounds

Hydroxamic compounds of formula (I) can be obtained in a variety of ways. The General scheme of vklyuchyenii.

The compounds of formula (I) is easily obtained from compounds of formula (A), R2amino acids, R22-halogen-substituted esters and the like. In compound A Y is preferably amino, and the connection is subjected to interaction with the compound B, where Y represents a halogen or a suitable leaving group. When connecting A Y is halogen, specialist immediately clear that compound B has a Y in the form of amino. When R1and R2do not form a single chain, R1fragment (B) is injected using traditional methods. For example, when using 2-halogenetor R1primary aminosidine under basic conditions replaces the halide or, if using the amino acid, it can be processed R1carbonyl compound such as aldehyde, after which the hydroxy fragment can be recovered by traditional methods of obtaining connection C. the compounds of formula A can be produced from known amino acids, including the 20 common amino acids, their derivatives (e.g., sarcosomataceae, 2-aminobutyric acid, pipecolinate acid and the like) or any such d-amino acids. Many of them are known or commercially available from, for example,necessaty can be obtained by any of many ways, known in the field of chemistry.

When more profitable is to obtain compounds of formula I using a halogen-substituted complex ether or halogen-substituted acids, such halogenated and halogenate known in the field of chemistry or can be obtained by known methods (see, for example, March, Advanced Organic Chemistry, Wiley Interscience).

Connection R3R4POZ receive standard methods. For example, PCl3can be alkylated and/or allerban education RPCl2or R2RPCl and then short-chain processed by alkanols with the formation of R3R4POZ.

In accordance with another variant, when R3and R1form a ring, the connection XC(O)CHR2NH2can be subjected to reaction under standard conditions with the formation of XC(O)CHR2NA(R1R3)POCl, which then closes with education:

< / BR>
(R1R preferably represents oxymethylene or oksietilenom.

When R4represents a heterocycle, methods of obtaining it gostinichnyh or postonline derivatives known in the field of chemistry. Preferred heterocyclic radicals4include 2 - or 3-thienyl, 2 - or 3-furyl, 2-the standard methods chemistry phosphoramidon, such as processing the amine phosphorylchloride in an inert solvent, and the like.

Usually hydroxamic acid is obtained in its final form at the final stage by treatment with hydroxylamine by well-known methods.

These stages can be modified to increase the yield of the target product. A qualified specialist will also be clear that the common choice of reagents, solvents and temperatures is an important component of a successful synthesis. Determination of optimal conditions, etc. is commonplace, but it is clear that a variety of compounds can be obtained in a similar manner, guided by the above scheme.

The source materials used to produce compounds of the present invention, known, can be obtained by known methods or are commercially available.

It is clear that a qualified expert in the field of organic chemistry can easily perform standard conversions of organic compounds without additional guidance, that is, the implementation of such transformations is fully within the competence of a specialist. These changes include (but are not limited to) restore the W, aromatic substitution (electrophilic and nucleophilic), etherification, esterification and saponification and the like. Examples of these transformations are described in the standard literature, such as March, Advanced Organic Chemistry (Wiley), Carey and Sundberg, Advanced Organic Chemistry (volume 2) and Keeting, Heterocyclic Chemistry (all 17 volumes).

The specialist can easily understand that some reaction is better to carry out the protection of other functional groups in the molecule, in order to avoid undesirable side reactions and/or to increase the yield of the reaction. To obtain such high outputs or to avoid unwanted reactions, the specialist often uses protective group. These reactions are described in the literature and well known to the skilled technician. Examples of many such transformations can be found, for example, in T. Green, Protecting Groups in Organic Synthesis. Of course, used as raw materials amino acids with reactive side chains preferably block (protect) to prevent unwanted side reactions.

Compounds of the present invention can have one or more chiral centers. As a result, it is possible to selectively receive one optical isomer (including vastly, or you can get both stereoisomer or both optical isomers (including diastereoisomers and enantiomers) simultaneously (racemic mixture). Since the compounds of the present invention can exist as racemic mixtures, mixtures of optical isomers (including diastereoisomers and enantiomers) or stereoisomers can be separated by known methods such as a method of chiral salts, method of chiral chromatography and the like.

In addition, it is clear that one optical isomer (including diastereoisomer-and enantiomer or stereoisomer may be more suitable properties than the other. So clearly understood that when in the description and the claims disclosed only racemic mixture, the disclosed and claimed both optical isomers (including diastereoisomers and enantiomers), or a stereoisomer, essentially separate from each other.

Applications

Metalloprotease (MP), found in the body, are involved in the destruction of the intercellular matrix containing intercellular proteins and glycoproteins. These proteins and glycoproteins play an important role in maintaining the size, shape, structure and stability of the tissue in the body. Inhibitorof. It is known that MP closely involved in remodelirovania tissue. As a result of such actions they, as has been said, take an active part in many of the violations that entail one of two things:

- tissue destruction, including degenerative diseases such as arthritis, multiple sclerosis and the like; metastasis or variability of tissue in the body;

- remodeling of tissue, including fibrotic disease, scarring, benign hyperplasia, and the like.

Compounds of the present invention treats disorders, diseases and/or unwanted conditions, which are characterized by undesirable or excessive activity of the proteases of this class. For example, the compounds can be used to inhibit proteases that

- break down structural proteins (i.e. proteins, providing stability and tissue structure);

- interfere with intercellular and intracellular signal transduction, including those involved in the regulation of cytokines, and/or processing of cytokines, and/or inflammation, tissue destruction and other diseases [Mohler KM, et al, Nature 370 (1994) 218-220, Gearing AJH, et al. Nature 370 (1994) 555-557, McGeehan GM, et al. Nature 370 (1994) 558-561], and/or contribute to the processes that are nieletni etc.

When used in this description, the expression "breach connected with the MP or disease associated with the MP" means the participation of unwanted or increased activity of the MP in the biological manifestation of diseases or disorders, in biological cascade leading to the violation, or in the form of symptom of disturbance. This "participation" MP includes:

- undesirable or increased activity as a "cause" of the violation or biological manifestations, regardless of cause increased activity of genetically, by infection, autoimmune reaction, trauma, biomechanical causes, lifestyle [e.g., obesity] or any other cause;

- MP as part of the observable manifestations of the disease or disorders. That is, the degree of disease or disorders can be identified by increased activity of the MP or, from a clinical point of view, undesirable, or elevated levels of MP testify about the disease. MP should not be considered a "hallmark" of the disease or disorder; unwanted or increased activity of the MP is part of the biochemical or cellular cascade that leads to the disease or violation of, or connected with him. In this regard, the inhibition of the activity of the MP interrupts Kano. So MP in tissues with a pronounced their localization is often specific to these tissues. For example, the distribution of metalloprotease involved in the process of tissue destruction in the joints, different from the distribution of metalloprotease found in other tissues. Therefore, although it is insignificant in relation to the activity or effectiveness of, any violation is preferably treated with compounds that act on specific MP found in infected tissues or body parts. For example, a connection exhibiting a higher degree of affinity and inhibition relative to MP, found in the joints (e.g., chondrocytes), it would be preferable for the treatment of diseases found there, in comparison with other compounds that are less specific.

In addition, some inhibitors are more bio-available for certain fabrics than others, and therefore the correct choice of the inhibitor with selectivity described above, provides specific treatment of disorders, diseases or unwanted conditions. For example, the compounds of the present invention with different abilities to penetrate into the Central nervous system. Therefore, the connection can be selected with you.

Determination of the specificity of an inhibitor of a particular MP is within the competence of the person skilled in the art. Suitable conditions analysis can be found in the literature. Known, in particular, methods of analysis for stromelysin and collagenase. For example, in U.S. patent N 4743587 refers to the method of Cawston, et al., Anal. Biochem. (1979) 99:340-345. The use of synthetic substrate in the analysis described in Weingarten, N. et al., Biochem. Biophy. Res. Comm. (1984) 139:1184-1187. You can, of course, to use any standard method for analysis of cleavage of structural proteins by metalloproteases. The ability of the compounds of the present invention to inhibit the activity of metalloprotease you can, of course, be checked by analysis methods found in the literature, or their variants. To confirm the inhibitory activity of the compounds of the present invention can use a dedicated metalloprotease or crude extracts, contains a number of enzymes that can destroy the fabric.

Due to their inhibitory action on MP compounds of the present invention is also useful for the treatment of the following disorders caused by the activity of metalloprotease.

Compounds of the present invention is also useful for p is whether pharmacology. For qualified specialist it is obvious that the preferred routes of administration depend on the state of disease that should be treated, and the selected dosage forms. Preferred methods of systemic injections include oral or parenteral administration.

However, the expert will easily understand the advantage of introducing inhibitor MP directly into the affected area for many disorders. For example, reasonable to introduce inhibitors MP directly in the affected area, such as area of surgical injury (for example, plastic surgery on vessels, i.e., angioplasty), area of scarring or burns (e.g., topically to the skin).

Because bone remodeling involves the process of metalloprotease, the compound of the present invention is useful for preventing looseness of the prosthesis. It is known that over time dentures start to hang out, to cause pain and can lead to further damage to the bones, which requires their replacement. Need a replacement of the prosthesis occurs for joint prostheses (e.g., prosthesis, hip, knee and shoulder joints), dental prostheses, including dentures, bridges and dentures, zakrepleniye system (for example, in congestive heart failure). It was assumed that one of the reasons angioplasty is higher than might be expected, the degree of prolonged failure (excessive time overlap), and that the activity of the MP becomes unwanted or increases in response to something that can be recognized by the body as damage the basement membrane of the vessel. Therefore, regulation of the activity of the MP with such indications as deletirovanie cardiomyopathy, congestive heart failure, atherosclerosis, plaque rupture, reperfusion injury, ischemia, chronic obstructive pulmonary disease, restenosis after angioplasty and aortic aneurysm, you may increase the duration of success of any other treatment or may be therapeutic in itself.

When skin care MP participate in the remodeling or the renewal of the skin. Therefore, regulation of MP improves the treatment of skin conditions, including (but not limited to removal, regulation and prevention of wrinkles and remedying damage to the skin by ultraviolet rays. This treatment includes prophylactic treatment or treatment before the appearance of obvious physiological manifestations. For example, the MP can be is me or after exposure to prevent or minimize the destruction after irradiation. In addition, the CHM participated in the violations and skin diseases associated with abnormal tissues resulting from abnormal update, in which take part metalloprotease, such as congenital bullous bullosa, psoriasis, scleroderma, and atopic dermatitis. Compounds of the present invention is also useful for the treatment of the effects of "normal" skin damage including scarring or "reduction" of the fabric, for example, after burns. Inhibition of MP is also useful in surgical operations on the skin to prevent scarring and promote normal growth of tissue in cases, for example, the replantation of limbs and refractory surgery (laser or cutting).

In addition, the CHM is linked to disorders including disordered remodeling other tissues, such as bone, such as otosclerosis and/or osteoporosis, or for specific organs) such as cirrhosis of the liver and pneumovirus. Similarly, when such diseases as multiple sclerosis, MP can be involved in disordered modeling of blood-brain barrier and/or myelin sheaths of nerve tissue. Therefore, the regulatory activity of MP mo

Apparently, the MP also participate in many infections, including cytomegalovirus [CMV, CMV) retinitis, HIV (HIV) and the resulting syndrome AIDS (AIDS).

MP can also participate in excessive vascularization, when the surrounding tissue must be destroyed in order to give the opportunity to form new blood vessels, such as angiofibroma and hemangioma.

Since MP break down the extracellular matrix, it can be assumed that inhibitors of these enzymes can be used as a means of birth control, providing, for example, preventing ovulation, preventing the penetration of sperm into and through the extracellular environment of the egg and implantation of the fertilized ovum and prevent the maturation of sperm.

In addition, they are useful for the prevention or cessation of premature labour and delivery.

As MP are involved in inflammatory reactions and in the processing of cytokines, the compounds are also useful as anti-inflammatory drugs for use in treatment of diseases with predominant inflammation, including inflammatory bowel disease, Crohn's disease, ulcerative colitis, pancreatitis, divertigo is when the cause of the disease is an autoimmune reaction, the immune response is often initiates the activity of the MP and cytokines. Regulation of CHM in the treatment of such autoimmune disorders is a useful method of treatment. Thus, inhibitors MP can be used to treat disorders, including lupus erythematosus, ankylosing spondylitis and autoimmune keratitis. Sometimes the side effects of therapy of autoimmunity lead to aggravation of other conditions mediated MP, and then spend therapies MP

inhibitors, for example, when the fibrosis caused by therapy of autoimmunity.

In addition, for this type of therapy is suitable and other fibrotic diseases, including pneumonia, bronchitis, emphysema, cystic fibrosis, acute respiratory distress syndrome (in particular, ostrofsky reaction).

When MP are involved in unwanted tissue destruction by external means, it can be treated with inhibitors MP. For example, they are effective as an antidote to the bite of a rattlesnake, as protivopravnoe tool in the treatment of allergic inflammation, septicemia and shock. In addition, they are useful as antiparasitic tools (e.g., malaria) and anti-infective funds. For example, they, apparently, are useful for the treatment of viral infection in the HIV-infection and AIDS.

It is assumed that MP inhibitors useful for the treatment of Alzheimer's disease, amyotrophic lateral sclerosis (ALS), muscular dystrophy, diabetic complications, in particular including the loss of viability of the tissue, coagulation, homologous disease (graft versus host), leukemia, cachexia, anorexia, proteinuria and, probably, the regulation of hair growth.

For some diseases, conditions or disorders inhibition MP presumably is the preferred method of treatment. Such diseases, conditions or disorders include arthritis (including osteoarthritis and rheumatoid arthritis), cancer (in particular, the prevention or relief of growth and metastasis of tumors), ocular disorders (in particular, corneal ulceration, poor healing of the cornea, macular degeneration and Parigi) and gum disease (in particular, periodontal disease and gingivitis).

Compounds of preferred (but non-derogable) for the treatment of arthritis (including osteoarthritis and rheumatoid arthritis), are those compounds that are selective towards metalloproteases and disintegrin-metalloproteases.

Compounds of preferred (but Neogen Auda those connections, which mainly inhibit gelatinase or collagenase type IV.

Compounds of preferred (but non-derogable this), for the treatment of eye disorders (in particular, corneal ulceration, poor corneal healing, macular degeneration and pterygia), are those compounds, which are widely inhibit metalloprotease. These compounds are preferably administered topically, preferably in the form of drops or gel.

Compounds of preferred (but non-derogable this) for the treatment of gum disease (in particular, periodontal disease and gingivitis), are those compounds, which mainly inhibit collagenase.

Songs

The compositions of the present invention contain:

(a) a safe and effective amount of the compounds of formula (I) and

(b) a pharmaceutically acceptable carrier. As described above, there are many diseases mediated by excessive or unwanted matrixarray activity metalloprotease. They include tumor metastasis, osteoarthritis, rheumatoid arthritis, skin inflammation, ulcers (particularly of the cornea), response to infection, periodontitis, and the like. Therefore, connect the ptx2">

The proposed connection can thus be made in the form of pharmaceutical compositions for use in treating or preventing these conditions. Use the standard technology of pharmaceutical preparations, such as described in Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., the most recent edition.

"Safe and effective amount" of the compounds of formula (I) is the quantity that is effective in the inhibition of metalloprotease in place (ahh) activity in a mammal subject, without undue adverse side effects (such as toxicity, irritation or allergic response) commensurate with a reasonable ratio of risk-to-benefit when used in accordance with the present invention. The specific "safe and effective amount" will obviously vary according to such factors as the particular condition that must be treated, the physical condition of the patient, duration of treatment, the nature of joint therapy (if any), the specific dosage form used by the media, the solubility therein of compounds of formula (I) and the required scheme dosing of the composition.

Also proposed is lovanii in this description, the term "pharmaceutically acceptable carrier" means one or more compatible solid or liquid fillers or substances for the manufacture of capsules, suitable for administration to a mammal. Used herein, the term "compatible" means that the components of the composition is able to mix with the proposed connection and with each other without any interaction that would substantially reduce the pharmaceutical efficacy of the composition under ordinary use conditions. Pharmaceutically acceptable carriers must, of course, be of sufficiently high purity and sufficiently low toxicity to render them suitable for administration to an animal, preferably a mammal undergoing treatment.

Some examples of substances which can serve as pharmaceutically acceptable carriers or components thereof are sugars, such as lactose, glucose and sucrose; starches, such as corn and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and methyl cellulose; powdered tragakant; malt; gelatin; talc; solid lubricants, such as stearic acid and magnesium stearate; calcium sulfate; vegetable oils, such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and oil of Theobroma; polyols, Tae, as Twins; wetting agents such as sodium lauryl sulfate; coloring agents; corrective substances; to contribute to a pelletizing; stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic saline and phosphate buffer solutions.

The choice of pharmaceutically acceptable carrier used in conjunction with the proposed connection is mainly determined by way of introduction connections.

If the proposed connection must be introduced by injection, the preferred pharmaceutically acceptable carrier is sterile, physiological saline (compatible blood suspendium agent), pH is increased to approximately 7.4.

In particular, pharmaceutically acceptable carriers for systemic injections include sugars, starches, cellulose and its derivatives, malt, gelatin, talc, calcium sulfate, vegetable oils, synthetic oils, polyols, alginic acid, phosphate buffered solutions, emulsifiers, isotonic saline, and pyrogen-free water. Preferred carriers for parenteral administration include polyethylene glycol, etiloleat, pyrrolidone, ethanol and sesame oil. Pharmaceuticle about 90% by weight of the entire composition.

The compositions of the present invention is preferably made in the form of dosage forms. In this description, the term "dosage form" is a composition of the present invention containing the compound of formula (I) in a quantity appropriate, according to medical practice, for the introduction of an animal, particularly a mammal, in the form of single (single) dose. These compositions preferably contain from about 5 to 1000 mg, preferably from 10 to 500 mg and more preferably from 10 to 300 mg of the compounds of formula (I).

The compositions of the present invention can be obtained in any of a variety of forms suitable for oral, rectal, local, nasal or parenteral administration. Depending on the desired specific method of administration can be used a variety of pharmaceutically acceptable carriers known in the art. They include solid or liquid fillers, diluents, hydrotropes, surface-active substances and substances for encapsulation. Can be entered as an optional pharmaceutically active substances, essentially no effect on the inhibitory activity of the compounds of formula (I). Number of media, is injection at a single dose of a compound of formula (I). Methods and compositions for making dosage forms suitable for methods of the present invention, are described in the following reference materials, which are all included in this description by reference: Modern Pharmaceutics, chapters 9 and 10 (the publisher of Banker &Rhodes, 1979); Lieberman et al. Pharmaceutical Dosage Forms, Tablets (1981); and Ansel, Introduction to Pharmaceutical Dosage Forms: 2nd edition (1976).

In addition to the compounds, the composition of the present invention contains a pharmaceutically acceptable carrier. When used in this description, the term "pharmaceutically acceptable carrier" means one or more compatible solid or liquid fillers or substances for the manufacture of capsules that are suitable for administration to a human or lower animal. Used herein, the term "compatible" means that the components of the composition is able to mix with the proposed connection and with each other without any interaction that would substantially reduce the pharmaceutical efficacy of the composition under ordinary use conditions. Pharmaceutically acceptable carriers must, of course, be of sufficiently high purity and sufficiently low toxicity to render them suitable for administration to a human or lower animal, protaminensulin holders or their components, are sugars such as lactose, glucose and sucrose; starches, such as corn and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethylcellulose and methyl cellulose; powdered tragakant; malt; gelatin; talc; solid lubricants, such as stearic acid and magnesium stearate; calcium sulfate; vegetable oils, such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and oil of Theobroma; polyols such as propylene glycol, glycerin, sorbitol, mannitol and polyethylene glycol; alginic acid; emulsifiers, such as Twins; wetting agents such as sodium lauryl sulfate; coloring agents; corrective substances; to contribute to a pelletizing; stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic saline and phosphate buffer solutions.

The choice of pharmaceutically acceptable carrier used in conjunction with the proposed connection is mainly determined by way of introduction connections.

If the proposed connection must be introduced by injection, the preferred pharmaceutically acceptable carrier is the bring about to 7.4.

Can be used in a variety of oral dosage forms, including such solid forms as tablets, capsules, granules, and bulk powders. These oral forms contain a safe and effective amount, usually at least about 5%, and preferably from about 25% to about 50%, of compounds of formula (I). Tablets can be compressed, trituration, with intersolubility coating, sugar coating, film coating or multilayer and may contain suitable binders, lubricants, fillers, loosening substances, coloring, corrective substances substances providing flowability, and substances that contribute to melting. Liquid oral dosage forms include aqueous solutions, emulsions, suspensions, solutions and/or suspensions obtained from nishiuchi granules and effervescent preparations obtained from effervescent granules, containing suitable solvents, preservatives, emulsifying agents, suspendresume substances, diluents, podslushivala melting substances, dyes and corrigentov.

Pharmaceutically acceptable carrier suitable for the preparation of dosage forms for perorally compatible auxiliary substances as inert fillers, such as calcium carbonate, sodium carbonate, mannitol, lactose and cellulose; binding substances such as starch, gelatin and sucrose; disintegrating agents such as starch, alginic acid and crosscarmellose; lubricants such as magnesium stearate, stearic acid and talc. To improve the characteristics of flowability of the powder mixture can be used sliding substances, such as silicon dioxide. The appearance can be added colorants, such as dyes, FD&C Podslushivala and corrigentov, such as aspartame, saccharin, menthol, peppermint and fruit corrigentov are useful excipients for chewable tablets. Capsules typically contain one or more solid diluents described above. The selection of carrier components depends on secondary considerations like taste, cost, and stability during storage, which are not essential for the purposes of the present invention, and thus can easily be made by a specialist in this field of technology.

Oral compositions include liquid solutions, emulsions, suspensions and the like. Pharmaceutically acceptable carriers suitable for receiving tow, emulsions and suspensions include ethanol, glycerol, propylene glycol, polyethylene glycol, liquid sucrose, sorbitol and water. For typical suspension suspendresume substances include methylcellulose, sodium carboxymethyl cellulose, AVICEL RC-591, tragakant and sodium alginate, a typical wetting agents include lecithin and Polysorbate 80 and typical preservatives include methyl paraben and sodium benzoate. Oral liquid compositions may also contain one or more components, such as podslushivala, corrigentov and dyes described above.

Such compositions may also be coated with conventional methods and contain coverage, depending on pH or time, resulting connection is released in the gastrointestinal tract near the desired site of application or has a prolonged action. Such dosage forms typically include (but are not limited to) one or more components from the group consisting of azettftalat cellulose, polivinilatsetatftalat, phthalate of hydroxypropylmethylcellulose, ethyl cellulose, Andreyevich coatings, waxes and shellac.

The compositions of the present invention can optionally include other active lekarstvennij, include sublingual dosage form, for insertion into the mouth for the insertion into the nose. Such compositions typically contain one or more soluble fillers, such as sucrose, sorbitol and mannitol, and binders, such as Arabian gum, microcrystalline cellulose, carboxymethyl cellulose and hypromellose. They may also include sliding substances, lubricants, podslushivala, dyes, antioxidants and corrigentov described above.

The compositions of the present invention can also enter the subject topically, for example, by direct overlay on epidermal or epithelial tissue of the subject or smearing it or percutaneous using the patch. Such compositions include, for example, lotions, creams, solutions, gels and solids. These local substance preferably contain a safe and effective amount of at least about 0.1%, and preferably about 1 to 5% of compounds of formula (I). Preferred carriers for the local introduction remain on the skin in the form of a solid film and resist removal when sweating or immersed in water. Usually the media is inherently organic and sposabella softeners, emulsifiers, thickeners, solvents or similar substances.

The routes of administration

In accordance with the present invention provides methods of treating or preventing disorders associated with excessive or unwanted activity of the matrix metalloprotease a person or animal by introducing him safe and effective amount of the compounds of formula (I). When used in this description to "violation associated with excessive or unwanted activity of the matrix metalloprotease" represents any violation, characterized by the splitting of proteins. The methods claimed in the present invention is suitable for treatment of such disorders, such as osteoarthritis, periodontal disease, corneal ulceration, tumor spreading and rheumatoid arthritis.

The compounds of formula (I) and compositions of the present invention can be injected locally or systemically. System introduction includes any method of introducing the compounds of formula (I) in the body tissue, such as intra-articular (in particular, for the treatment of rheumatoid arthritis), vnutriobolochechnoe, epidural, intramuscular, percutaneous, intravenous, intraperitoneal, subcutaneous, sublingual, rectal and peroral The specific dosage of the injected inhibitor, as well as the duration of treatment, whether local or systemic, interdependent. Dosing schedules and treatment also depend on such factors as specifically used the compound of formula (I), the indication for treatment, the ability of the compounds of formula (I) to achieve a minimum inhibitory concentration in the place where you want to inhibit the matrix metalloprotease, personal performance of a subject (such as weight), the ability of adherence to treatment regimens, and the presence and severity of side effects of treatment.

Usually an adult (weighing about 70 kg) is administered per day with the systemic administration of from about 5 to 3000 mg, preferably from about 5 to 1000 mg, and more preferably from 10 to 100 mg of the compounds of formula (I). It is clear that these limits dosing is given only as an example and that the daily introduction can be adjusted depending on the above factors.

The preferred method of administration for treatment of rheumatoid arthritis is oral administration or parenteral administration by intra-articular injection. As we know from theory and practice of medicine, all preparations for parenteral administration must be sterile. For mammals, in private IMEMO from 10 to 1000 mg.

The preferred method of system introduction is oral administration. Preferred is the individual dose from about 10 to 1000 mg, preferably from 10 to 300 mg.

Local injection can be used for systemic delivery of the compounds of formula (I) or for local treatment of the subject. The amount of the compounds of formula (I) for local administration depends on such factors as skin sensitivity, type and location of tissue to be treated, the input composition and carrier (if any), specifically the insertion compound of the formula (I), as well as the specific disorder and the required degree distribution system (as distinguished from local) effects.

The inhibitors of the present invention can be targeted at specific places where the accumulated matrix metalloprotease, by applying the targeting ligands. For example, to target inhibitors became the matrix metalloprotease, in tumor inhibitor kongugiruut with the antibody (or fragment), which is immunoreactive against the tumor marker, as it is usually understood in relation to obtaining immunotoxins. The target ligand may be a ligand suitable for Modestum with a token for the intended target tissue. Methods of binding compounds of the present invention with the target ligand are well known and similar to those described below for binding with the carrier. The conjugates are administered as described above.

For localized States preferred is a local introduction. For example, for treatment of ulcerated cornea can be used for direct application to the affected eye of the drug in the form of eye drops or spray. For the treatment of corneal compounds of the present invention can also be prepared in the form of gels or ointments, or can be incorporated into collagen or of the hydrophilic polymer protective device. The agents can also be introduced in the form of contact lenses or reservoir, or in the form of subconjunctival drug. For the treatment of skin inflammation connection is applied locally in a certain place in the form of a gel, paste or ointment. Treatment reflects, therefore, the nature of the state, and for any selected mode are suitable dosage form.

Of course, that in all the cases described above the compounds of the present invention can be introduced separately or in mixtures, and compositions can optionally contain one is about the present invention also inhibit bacterial metalloprotease, although usually at a lower level than metalloprotease mammals. Some bacterial metalloprotease seems less dependent on the stereochemistry of the inhibitor, whereas the ability of diastereomers deactivate protease mammals between them found significant differences. Thus, this feature of the activity can be used to establish differences between mammalian enzymes and enzymes of bacteria.

The preparation and use of antibodies

Compounds of the present invention can be used in the immunization schemes for obtaining antisera, immunospecific in relation to the compounds of the present invention. Because the proposed connection is relatively small, it is reasonable to associate them with antigenic neutral media, such as traditionally used media type hemocyanin KLH or serum albumin. For those of the proposed compounds, which have a carboxyl functional group, the binding medium may be carried out by methods already known in this field. For example, carboxyl residue may be restored to the aldehyde and attached to the carrier through Rea the m formed imine linkages. Carboxyl residue may be subjected to interaction with the side amino groups using condensing agents, such as dicyclohexylcarbodiimide, or other carbodiimide dehydrating agents.

To link, you can also use binding compounds; as homobifunctional, so heterobifunctional linking compounds are manufactured by Pierce Chemical, Rockford, III. Received immunogenic complex can then be injected with a suitable mammalian subjects such as mice, rabbits and the like. Appropriate schemes include multiple injection of the immunogen in the presence of adjuvants on the program, which promotes the production of antibodies in the serum. The titers of the immune serum can easily be measured by the methods of immunological analysis (now standard in this area) with compounds of the present invention as antigens.

The obtained antisera can be used directly or can be obtained monoclonal antibodies by collecting peripheral blood lymphocytes or spleen of immunized animal and the immortalization of cells producing antibodies with subsequent identification analnye or monoclonal preparations can then be used to control the modes of treatment or prevention using the compounds of the present invention. Suitable samples, such as samples obtained from blood, serum, urine or saliva, you can test for the presence of the injected inhibitor at various points during treatment, using standard methods of immunological analysis using containing antibody preparations of the present invention.

Compounds of the present invention can also be combined with labels such as scintigraphic labels, such as technetium-99 or I-131, using standard methods of binding. Labeled compound is administered to the subjects to determine the locations of excess amounts of one or more metalloprotease in vivo. The ability of inhibitors to selectively contact metalloproteases advantageous in this case because it allows you to map the distribution of these enzymes on site (in situ). This technique can also be used in histological procedures, and labeled compounds of the present invention can be used in comparative immunological assays.

The following examples are only for illustration compounds, compositions and applications of the present invention.

Obtaining compounds

Connections get so as phenylethylamine (6,63 ml, of 52.8 mmol) and triethylamine (7,39 ml, 53 mmol) in anhydrous N,N-dimethylformamide (80 ml) cooled to 0oC and the resulting mixture was added dropwise a solution of methylpropanoate (5 ml, of 52.8 mmol) in anhydrous N,N - dimethylformamide (40 ml). The reaction mixture is allowed to mix for 20 minutes at 0oC. the Mixture is poured into 250 ml ethyl acetate and washed with water (3 times), dried over sodium sulfate and evaporated to obtain a colorless oil. By dissolving the crude oil in 75 ml of ether to obtain the hydrochloride. Separately to 2.5 ml of methanol at 0oC added dropwise to 3.8 ml acetylchloride. This solution is added dropwise to an ethereal solution. Precipitated precipitated solids are collected by filtration to obtain 9.2 grams (76%) of methyl ester hydrochloride N-(2-phenethyl) glycine as colorless solid.

Methyl ester of N-(diphenylphosphinyl)-N-(2-phenylethyl) glycine: Diphenylphosphoryl (0,42 ml, 2.2 mmol) dissolved in dichloromethane (5 ml) and cooled to 0oC. To the resulting solution was added a solution of methyl ester of N-(2-phenethyl)glycine (500 mg, 2.2 mmol) and N-methylmorpholine (0,73 ml, 6.6 mmol) in dichloromethane (5 ml). The reaction mixture is stirred 16 hours at room temperature, washed with water and RA is ethyl) glycine as colorless solid.

N-Hydroxy-2-[[diphenylphosphine](2-phenylethyl)amino] ndimethylacetamide:

Methyl ester of N-(diphenylphosphinyl)-N-(2-phenylethyl) glycine (160 mg, 0.41 mmol) was dissolved in methanol (2.5 ml). To the resulting solution was added gidroxinimesoulid (57 mg, 0.81 mmol) and then 2 mmol of 25% methanol solution of sodium methoxide. The reaction mixture is stirred for 16 hours, neutralized 1 N. chloroethanol acid and concentrated. The crude product is purified by flash chromatography on silica gel with getting to 41.6 mg (26%) of N-hydroxy-2-[[diphenylphosphine] (2 - phenylethyl)amino]ndimethylacetamide in the form of a colorless solid: MC (IS): m/z 395 [M+N)+, 417 [M+Na]+, 433 [M+K]+. (R1= phenylethyl, R2= H, R3= phenyl, R4= phenyl).

Example 2

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Methyl ester of N-(methylphenylphosphinic)-N-(2-phenylethyl) glycine:

Methylphenylphosphinic (0.45 ml, 3,26 mmol) dissolved in dichloromethane (5 ml) and cooled to 0oC. To the resulting solution was added dropwise a solution of 750 mg (3,27 mmol) of methyl ester of N-(2-phenethyl)glycine and N-methylmorpholine (1.1 ml, 10 mmol) in dichloromethane (5 ml). The reaction mixture is stirred for 1 hour, stirring to room temperature. The mixture is diluted with ethyl acetate, the organic phase is washed parafii on silica gel to obtain methyl ester of N- (methylphenylphosphinic)-N-(2-phenylethyl) glycine as colorless oil.

N-Hydroxy-2-[[methylphenylphosphinic](2-phenylethyl)amino] ndimethylacetamide:

Methyl ester of N-(methylphenylphosphinic)-N-(2-phenylethyl) glycine (300 mg, 0,905 mmol) process of 0.77 ml of NH2OK (1,76 M in methanol, the solution obtained as described in Fieser and Fieser, vol 1, p 478). The mixture is stirred for 3 hours at room temperature, neutralized with formic acid and concentrated. The crude product is purified by flash chromatography on silica gel (ethyl acetate: ethanol=85:15) to obtain the product with minor impurities. Preparative TLC (ethyl acetate: ethanol= 90: 10) gives N-hydroxy-2-[[methyl-phenylphosphine](2-phenylethyl)amino]ndimethylacetamide in the form of a colorless solid: MS (IS):m/z 333 [M+N)+, 350 [M+NH4]+, 355 [M+Na]+, 371 [M+K]+. (R1= phenylethyl, R2= H, R3= methyl, R4= phenyl).

Example 3

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Benzyl ether of D-leucine, D-leucine (10 g, 76,23 mmol) suspended in benzyl alcohol (157 ml) and heated to 55oC. After the mixture was bubbled with gaseous hydrogen chloride and the reaction mixture becomes very viscous. Then add 150 ml of benzene under vigorous stirring. After 30 minutes of ozonation by heating the diluted mixture and mixing is facilitated. The reaction mixture is continued amaut and the reaction mixture is allowed to mix at 55oC for another 30 minutes. Allow the reaction mixture to cool to room temperature and dilute it with ethyl acetate. The product is extracted using 1 M HCl (3 times) and remove organics in the waste. Wash water are combined to pH 8 with 50% aqueous NaOH solution. Several times the amine is extracted with ethyl acetate. The organics dried over sodium sulfate and evaporated. To the obtained oil was added ether and bubbled with HCl deposition benzyl ether-D-leucine in the form of chloroethanol salt.

Benzyl ester of N-benzyl-D-leucine, Methyl ester of D-leucine (3 grams 11,66 mmol) is dissolved in methanol and the resulting solution was added sodium acetate (1.9 g, with 23.3 mmol) and then benzaldehyde (1.2 ml, 11,66 mmol). Mixture is allowed to mix for 10 minutes, then added dropwise a solution of lamborginid sodium (427 mg, 6.8 mmol) in methanol (4 ml). The reaction mixture was stirred for 3 hours, after which TLC determine completion of the reaction. To the reaction mixture is added with stirring 10% aqueous solution of NaHCO3. Then remove volatile substances and the product extracted with ether (3 times). Organics washed with water (2 times), dried over sodium sulfate and evaporated to obtain benzyl ester N-benzyl-D-leucine in the form of bescot the ID (0,89 ml, 6.42 per mmol) was dissolved in dichloromethane and cooled to 0oC. To the resulting solution was added a solution of methyl ester of N-benzyl-D-leucine (2 g, 6.42 per mmol) and N-methylmorpholine (1.5 ml, 13,48 mmol) in dichloromethane. Added a catalytic amount of 4-dimethylaminopyridine and allow the reaction mixture to mix for 22 hours. The reaction mixture was concentrated and to the residue is added ethyl acetate. The resulting mixture was washed with water and brine, dried over sodium sulfate and concentrated. By flash chromatography on silica gel (100% ethyl acetate) was isolated diastereomers.

N-((R/S)-Methylphenylphosphinic)-N-benzyl-D-leucine: Flask containing benzyl ester N-(R/S-methylphenylphosphinic)-N-benzyl-D - leucine (2,04 g, a 4.53 mmol) and 10% Pd/C (500 mg), vaccum and then give it the methanol. Create an environment of hydrogen and allow the reaction mixture to mix for 45 minutes. The mixture is filtered through celite and the filtrate collected and concentrated to obtain N-((R/S)-methylphenylphosphinic)-N-benzyl-D-leucine as a white glassy substance.

N-Benzyloxy-2-(R)-[[(R)-methylphenylphosphinic]benzylamino]-4 methylpentane and N-benzyloxy-2-(R)-[[(S)-methylphenylphosphinic] benzylamino]-4-methylpentanoic: N-((R/S)-methylphenylphosphinic)-N-benzyl-D added hydroxybenzotriazole (1,69 g, 12.5 mmol), N-methylmorpholin (1,37 ml, 12.5 mmol) and 1-ethyl-N-(3 - dimethylaminopropyl)carbodiimide (EDAC, 959 mg, 5 mmol). After stirring for 10 minutes, add O-benzylhydroxylamine hydrochloride (666 mg, of 4.17 mmol) and allow the reaction mixture to mix for 3 hours with warming to room temperature. TLC shows the presence of two diastereomers. Add water to the mixture and extracted with a mixture of ethyl acetate. Organic unite washed with water and brine, dried over sodium sulfate and concentrated to obtain oil. Then allocate the diastereomers by flash chromatography on silica gel (hexane: ethyl acetate= 1: 1) to give N-benzyloxy-2-(R)-[[(R) -methylphenylphosphinic] benzylamino] -4-methylpentanoic: Rf= 0,25 (hexane:ethyl acetate=1:1);31P NMR (CD3OD) d 43,89; and N-benzyloxy-2-(R)-[[(S)-methylphenylphosphinic]benzylamino] -4-methylpentanoic: Rf=0,15 (hexane:ethyl acetate - 1:1).

N-Hydroxy-2-(R)-[[(S)-methylphenylphosphinic]benzylamino]- 4-methylpentanoic: N-Benzyloxy-2-(R)-[[(S)- methylphenylphosphinic]-benzylamino]-4-methylpentanoic (334 mg, 0,719 mmol) and 80 mg of 10% Pd/C vacuum in the flask. Add 10 ml of methanol and injected into the flask hydrogen. The reaction mixture is allowed to mix at room temperature for 2 hours, POWAY to get a white glassy solid. The product is dissolved in ethyl acetate and the resulting solution was added dropwise hexane to drop product into the sediment, resulting in a gain of N-hydroxy-2-(R)- [[(S)-methylphenylphosphinic]benzylamino]-4-methyl-pentanone in the form of a colorless solid: MC (IS): m/z 375 [M+N)+, 397 [M+Na]+, 413 [M+K]+.

N-Hydroxy-2-(R)-[[(R)-methylphenylphosphinic] benzylamino] -4 - methylpentanoic: N-Benzyloxy-2-(R)-[[(R)-methylphenylphosphinic]- benzylamino]-4-methylpentanoic (460 mg, 0,99 mmol) and 10% Pd/C (100 mg) vacuum in the flask. Add methanol (10 ml) and injected into the flask hydrogen. The reaction mixture is allowed to mix at room temperature for 2 hours, followed by TLC indicates that the reaction is complete. The mixture is filtered through celite and the filtrate is collected and evaporated to obtain a white glassy solid. Hydroxamic acid is crystallized by dissolution in ethyl acetate and addition of hexane dropwise until the solution becomes turbid. The result is N-hydroxy-2-(R)-[[(R)-methylphenylphosphinic] benzylamino]-4-methylpentanoic in the form of a white crystalline solid: MS (IS): m/z 375 [M+H)+, 397 [M+Na]+, 413 [M+K]+. (R2= isobutyl, R1= benzyl, R3= methyl, R4oC. To the resulting solution was added dropwise thionyl chloride (25,5 ml, 350 mmol). The reaction mixture is allowed to mix at room temperature for 16 hours after which remove volatile substances from getting not quite white solid. The product is recrystallized from a mixture of ethyl acetate-methanol to obtain methyl ester hydrochloride, D-leucine in the form of a friable white solid.

Methyl ester of N-benzyl-D-leucine: Hydrochloride of the methyl ester of D-leucine dissolved in methanol. To the solution was added sodium acetate (39,4 g, 480 mmol) and then benzaldehyde (to 19.8 ml, 195 mmol). The resulting mixture is stirred for 15 minutes and add to it for 15 minutes a solution of lamborginid sodium (7,1 g, 113 mmol) in methanol (50 ml). After 3 hours the reaction ends. Add with stirring a 10% aqueous sodium bicarbonate solution and after 10 minutes remove volatile substances. The product is extracted with ether and washed with water (2 times). The ester mixture is dried over sodium sulfate and evaporated to obtain and 39.9 g of methyl ester of N-benzyl-D-leucine in the form of a colorless oil.

Methyl ester of N-(dimethylphosphino)-N-benzyl-D-leucine: Dimethylphosphoric (200 mg, 1.78 mmol) dissolved in dichloromethane ( mg, a 1.75 mmol) and N-methylmorpholine (of 0.44 ml, 4 mmol) in dichloromethane (5 ml). Added a catalytic amount of 4-dimethylaminopyridine and give the reaction mixture is mixed for 16 hours at room temperature.

Then filter out the solids and the filtrate is collected and evaporated. The crude product is purified by flash chromatography on silica gel (ethyl acetate:methanol=96:4) to give the methyl ester of N-(dimethylphosphino)-N-benzyl-D-leucine in the form of a colorless solid.

N-Hydroxy-2(R)-[[dimethylphosphine]benzylamino]-4-methyl-pentanone:

Methyl ester of N-(dimethylphosphino)-N-benzyl-D-leucine (158 mg, 0.51 mmol) is treated with a solution of NH2OK (2.8 ml, 1,76 M in methanol) obtained as described in Fieser and Fieser, vol 1, p 478. Give the reaction mixture is mixed for 3 hours at room temperature, after which by TLC indicates that the reaction has ended. The reaction mixture is neutralized 1 M aqueous HCl solution to remove volatiles to obtain oil. Add methanol and then water is added dropwise until the solution becomes turbid. Collect by filtration the crystals, which gives N-hydroxy-2(R)-[[dimethylphosphine]benzylamino]-4-methylpentanoic as bescoto the3= methyl, R4= phenyl).

Example 5

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< / BR>
Methyl ester of N-benzyl-D-alanine, Methyl ester of D-alanine (4 g, 28,66 mmol) dissolved in methanol (100 ml). To the solution was added sodium acetate (5,88 g, 71,65 mmol) and benzaldehyde (2,9 ml, 28,66 mmol). The resulting mixture is stirred for 15 minutes and added to it dropwise a solution of lamborginid sodium (1.08 g, and 17.2 mmol) in methanol (5 ml). After stirring for 2 hours, evaporated under reduced pressure, methanol and the product is extracted with ether and washed with water (2 times). The crude product is purified by flash chromatography on silica gel (hexane-ethyl acetate=8:2) to obtain 3.3 g of methyl ester of N-benzyl-D-alanine in the form of butter.

Methyl ester of N-((R)-methylphenylphosphinic)-N-benzyl-D-Alcina:

Methylphenylphosphinic (361 mg, 2,07 mmol) dissolved in dichloromethane (2.5 ml) and cooled to 0oC. To the resulting solution was added a solution of methyl ester of N-benzyl-D-alanine (400 mg, 2,07 mmol) and N-methylmorpholine (0.51 ml, 4.6 mmol) in dichloromethane (2.5 ml). To a stirred mixture was added a catalytic amount of 4-dimethylaminopyridine. The reaction mixture is stirred for 16 hours at room temperature, washed with water and brine, dried over butilatsetat) to give the methyl ester of N-((R)-methylphenylphosphinic)-N-benzyl-D - alanine in the form of butter.

N-Hydroxy-2(R)-[[(R)-methylphenylphosphinic]benzylamino] propionamide: Methyl ester of N-((R)-methylphenylphosphinic)-N-benzyl - D-alanine (181 mg, 0.55 mmol) is treated with a solution of NH2OK (2.2 ml, 1,76 M in methanol) obtained as described in Fieser and Fieser, vol 1, p 478. The reaction mixture is stirred for 16 hours, followed by TLC indicates that the reaction has ended. The reaction mixture is neutralized 1 M aqueous solution of HCl and remove volatile substances. The target product was then purified on flash silica using THF as eluent. The resulting residue is crystallized by dissolution in ethyl acetate and addition of hexane until the turbidity of the solution. The result is N-hydroxy-2(R)-[[(R)-methylphenylphosphinic]benzylamino] propionamide in the form of a solid and dense colorless crystals: MC (IS): m/z 333 [M+H]+, 355 [M+Na]+. (R1= benzyl, R2= methyl, R3= phenyl, R4= phenyl).

Example 6

< / BR>
Methyl ester of N-(diphenylphosphinyl)-N-benzyl-D-alanine: Diphenylphosphoryl (0.2 ml) dissolved in dichloromethane (5 ml) and cooled to 0oC. To the resulting solution was added a solution of methyl ester of N-benzyl-D-alanine (243 mg, of 1.26 mmol) and triethylamine (of 0.39 ml, 2.8 mmol) in dichloromethane (2.5 ml). To react is for 48 hours at room temperature. The dichloromethane solution was diluted with 20 ml dichloromethane and then washed with 1 M aqueous HCl solution (2 times). The product was then purified by flash chromatography on silica gel (ethyl acetate: hexane=8:2) to give the methyl ester of N-(diphenylphosphinyl)-N-benzyl-D-alanine in the form of butter.

N-Hydroxy-2(R)-[[diphenylphosphine]benzylamino] propionamide: methyl ether N-(diphenylphosphinyl)-N-benzyl-D - alanine (100 mg, 0.25 mmol) was added a solution of NH2OK (0,88 ml, 1,76 M in methanol) obtained as described in Fieser and Fieser, vol 1, p 478. The reaction mixture is stirred for 16 hours, followed by TLC indicates that the reaction has ended. The reaction mixture is neutralized 1 M aqueous solution of HCl and remove volatile substances. The product was then purified by flash chromatography on silica gel (100% ethyl acetate) to obtain N-hydroxy-2(R)-[[diphenylphosphine] benzylamino]propionamide in the form of an oil: MS (IS): m/z 395 [M+H]+, 417 [M+Na]+. (R1= benzyl, R2= methyl, R3= phenyl, R4= phenyl).

Example 7

< / BR>
< / BR>
Methyl ester of N-(3-picolyl)-D-leucine: Hydrochloride of the methyl ester of D-leucine (20 g, 110,43 mmol) dissolved in methanol. To the resulting solution was added sodium acetate (22,64 g, 276 mmol) and then 3-pyridinemethanol added within 15 minutes lambrogini sodium (4.15 g, 66 mmol). After stirring for 16 hours at room temperature, the methanol is evaporated under reduced pressure and the oil obtained is dissolved in ethyl acetate and washed with water (2 times). The organics dried over sodium sulfate and concentrated to oil. The product was then purified by flash chromatography on silica gel (100% ethyl acetate) to obtain methyl ester of N-(3-picolyl)-D-leucine in the form of butter.

N-Hydroxy-2(R)-[[(R/S)-methylphenylphosphinic] -3 - picolylamine] -4-methylpentanoic: Methylphenylphosphinic (12,23 g, 70 mmol) dissolved in dichloromethane (100 ml) and cooled to 0oC. To the resulting solution was added a solution of methyl ester of N-(3-picolyl)-D-leucine (15.5 g, 65,6 mmol) and N-methylmorpholine (19,24 ml, 175 mmol) in dichloromethane (100 ml). Added a catalytic amount of 4-dimethylaminopyridine and give the reaction mixture is mixed for 16 hours at room temperature. Add additional methylphenylphosphinic (2 g, 11,46 mmol). The reaction mixture is further mixed for 24 hours, after which the reaction ends. TLC does not provide separation of the diastereomers. The product was then purified by flash chromatography on silica gel (ethanol:ethyl acetate=5:95) to give the methyl ester of N-((R/S)-were, who must register as described in Fieser and Fieser, vol 1, p 478. The reaction mixture is stirred for 16 hours, followed by TLC indicates that the reaction has ended. The reaction mixture is neutralized 1 M aqueous solution of model HC1 and remove volatile substances. The product was then purified by flash chromatography on silica gel (ethanol: ethyl acetate= 10: 90) to give 8.6 g of N-hydroxy-2(R)-[[(R/S)-methylphenylphosphinic]-3 - picolylamine]-4-methylpentanoate in the form of a mixture of diastereoisomers (60R/40S):MS (IS): m/z 376 [M+H]+, 398 [M+Na]+, (R1= 3-pyridylmethyl, R2= 2-isobutyl, R3= methyl, R4= phenyl).

Example 8

Scheme IV

< / BR>
Methyl ester of N-((R and S)-methylphenylphosphinic)-D-leucine: Methylphenylphosphinic (113 mg, of 0.65 mmol) dissolved in dichloromethane (5 ml) and cooled to 0oC. To the resulting solution was added a solution of methyl ester hydrochloride D-leucine (100 mg, 0.55 mmol) and N-methylmorpholine (of 0.18 ml of 1.65 mmol) in dichloromethane (3 ml). After stirring for 16 hours at room temperature on a plate for TLC see two spots. These compounds separated by flash chromatography on silica gel (ethyl acetate:methanol=95:5) to give two diastereomeric products: methyl ester of N-((R)-methylphenylphosphinic)-D-Lac tracecut).

N-Hydroxy-2(R)-[[(R)-methylphenylphosphinic] amino] -4 - methylpentanoic: Methyl ester of N-((R)-methylphenylphosphinic)-D - leucine (60 mg, 0.21 mmol) is treated with a solution of NH2OK (or 0.57 ml, 1,76 M in methanol) obtained as described in Fieser and Fieser, vol 1, p 478. The reaction mixture is stirred for 7 hours, followed by TLC indicates that the reaction is complete. The reaction mixture is neutralized 1 M aqueous solution of HCl and remove volatile substances. The residue is purified by flash chromatography on silica gel (ethyl acetate: ethanol = 95:5) to give N-hydroxy-2(R)-[[(R)-methylphenylphosphinic]amino]-4 - methylpentanoate in the form of a colorless solid: MS (IS): m/z 285 [M+H]+.

N-Hydroxy-2(R)-[[(S)-methylphenylphosphinic] amino] -4-methylpentanoic: Methyl ester of N-((S)-methylphenylphosphinic)-D-leucine (55 mg, 0,19 mmol) is treated with a solution of NH2OK (or 0.57 ml, 1,76 M in methanol) obtained as described in Fieser and Fieser, vol 1, p 478. The reaction mixture is stirred for 6 hours, followed by TLC indicates that the reaction is complete. The reaction mixture is neutralized 1 M aqueous solution of HCl and remove volatile substances. The residue is purified by flash chromatography on silica gel (ethyl acetate: ethanol =80:20) followed by crystallization from a mixture of De colorless solid: MS (IS): m/z 285 [M+H]+. (R1= H, R2= isobutyl, R3= methyl, R4= phenyl).

Example 9

Scheme V

< / BR>
The ethyl ethylvanillin: Mixture diethylphenylphosphine (4,5 22,70 mmol), ethyliodide (of 0.24 ml, 3 mmol) and benzene (100 ml) is stirred and heated at 85oC for 24 hours. TLC shows that the reaction was carried out on 30%. Add another portion of ethyliodide (0,30 ml, 3.75 mmol) and the reaction mixture is stirred for 36 hours at 85oC, after which time TLC shows that the reaction is complete. Removed on a rotary evaporator volatiles, resulting in getting the ethyl ethylvanillin in the form of butter.

Ethylenepropylene: To a solution of ethyl ethylvinylacetate (2 g, 10 mmol) in benzene (200 ml) was added oxalicacid (1.3 ml, 15 mmol). The mixture is stirred for 3 hours at room temperature. On a rotary evaporator to remove volatile substances and the product is dried in vacuum for 12 hours to obtain ethylphenethylamine in the form of butter.

Methyl ester of N-((R and S)-ethylvanillin)-N-benzyl-D - alanine: To a solution of ethylphenethylamine (1.04 g, 5.5 mmol) in dichloromethane (15 ml) was added a solution of methyl ester of N-benzyl-D-alanine (1,37 g, 7.1 mmol) and N-methylmorpholine (1,36 ml, 12.4 mmol) in dichlormethane room temperature. On the plate for TLC see two spots. These compounds separated by flash chromatography on silica gel (ethyl acetate:methanol=95:5) to give two diastereomeric products: methyl ester of N-((S)-ethylvanillin)-N-benzyl-D-alanine, Rf= 0,25 (100% ethyl acetate) and methyl ester of N-((R)-ethylvanillin)-N - benzyl-D-alanine, Rf= 0,35 (100% ethyl acetate).

N-Hydroxy-2(R)-[[(S)-ethylvanillin] amino] propionamide: Methyl ester of N-((S)-ethylvanillin)-N-benzyl-D-alanine (105 mg, 0.30 mmol) is treated with a solution of NH2OK {1.0 ml, 1,76 M in methanol) obtained as described in Fieser and Fieser, vol 1, p 478. The reaction mixture is stirred for 16 hours, followed by TLC indicates that the reaction is complete. The reaction mixture is neutralized 1 M aqueous solution of HCl and remove volatile substances. The residue is purified by flash chromatography on silica gel (ethyl acetate: methanol = 95:5) to give N-hydroxy-2(R)-[[(S)-ethylvanillin]amino] propionamide in the form of a colorless solid: MS (IS): m/z 347 [M+H]+369 [M+Na]+.

N-Hydroxy-2(R)-[[(R)-ethylvanillin] amino]propionamide:

Methyl ester of N-((R)-ethylvanillin)-N - benzyl-D-alanine (333 mg, 0.96 mmol) is treated with a solution of NH2OK (3,3 ml, 1,76 M is e which by TLC determine that the reaction is complete. The reaction mixture is neutralized 1 M aqueous solution of HCl and remove volatile substances. The residue is purified by flash chromatography on silica gel (ethyl acetate: methanol = 95: 5) to give 110 mg (33%) of N-hydroxy-2(R)-[[(R)-ethylvanillin] amino]propionamide in the form of a colorless solid: MS (IS): m/z 347 [M+H]+, 369 [M+Na]+. (R1= benzyl, R2= methyl, R3= ethyl, R4= phenyl).

Example 10

< / BR>
< / BR>
Methyl ester of N-hexyl-D-alanine, Methyl ester of D-alanine (1.5 g, of 10.75 mmol) is dissolved in 50 ml of methanol and cooled to 0oC. To the solution was added hexanal (1.3 ml, of 10.75 mmol) and then sodium acetate (2,62 g, 32 mmol). After stirring for 15 minutes at 0oC add lambrogini sodium (440 mg, 7 mmol) and the mixture is stirred for another 16 hours at room temperature. The methanol is evaporated and the resulting residue is dissolved in ether and transferred into a separating funnel, washed with water (2 times), dried over sodium sulfate and evaporated obtaining of 1.81 g of methyl ester of N-hexyl-D-alanine as a colourless oil.

Methyl ester of N-((R)-methylphenylphosphinic)-N-hexyl-D-alanine:

Methylphenylphosphinic (1,05 g, 6 mmol) dissolved in dichloromethane (50 ml) and the OHL is) and triethylamine (2.1 ml, 15 mmol) in dichloromethane (10 ml). Added a catalytic amount of 4-dimethylaminopyridine and the reaction mixture is stirred for 16 hours, washed with water and brine, dried over sodium sulfate and concentrated. The crude product is purified by flash chromatography on silica gel (100% ethyl acetate) to obtain methyl ester of N-((R)-were-phosphinyl)-N-hexyl-D-alanine in the form of butter.

N-Hydroxy-2(R)-[[(R)-methylphenylphosphinic]hexylamino] propionamide: Methyl ester of N-((R)-methylphenylphosphinic)-N-hexyl-D - alanine (198 mg, 0.61 mmol) is treated with a solution of NH2OK (2 ml, 1,76 M in methanol) obtained as described in Fieser and Fieser, vol 1, p 478. The reaction mixture is stirred for 16 hours, followed by TLC indicates that the reaction has ended. The reaction mixture is neutralized 1 M aqueous solution of HCl and remove volatile substances. The crude product is purified by preparative TCX (ethyl acetate: methanol= 95:5) to obtain 110 mg of N-hydroxy-2(R)-[[(R)-methylphenylphosphinic]hexylamino]propionamide in the form of a colorless solid: MS (IS): m/z 327 [M+N)+, 349 [M+Na]+. (R1= 2-hexyl, R2= methyl, R3= methyl, R4= phenyl).

In table. 1 presents additional examples of the what in or starting materials, obtained by known methods:

< / BR>
These examples are for professionals sufficient guidance for the implementation of the present invention and in no way limit the invention.

Examples of compositions and methods of use

Compounds of the present invention is suitable for the manufacture of compositions for the treatment of diseases and the like. The following examples of compositions and methods do not limit the invention, but gives the operator an indication of how to obtain and apply the compounds, compositions and methods of the present invention. In each case, the compounds of formula I may be substituted with, for example, the compound shown below, with similar results.

Examples of methods of application do not limit the invention, but are for the specialist guidance on how to apply the compounds, compositions and methods of the present invention. Qualified practitioners will be clear that the examples give direction and can be changed depending on the condition and the patient.

An example of A

Composition in tablet form for oral administration has the following composition, mg:

The compound of example 9 - 15

Lactose - 120

Maize is Y. the formula I, used, essentially, with the same results.

A woman weighing 60 kg (132 lb) suffering from rheumatoid arthritis, treated by the method according to the present invention. In particular, it has introduced tablet orally for 2 years under the scheme of three tablets per day.

At the end of the treatment period the patient was investigated and found that the inflammation decreased and increased mobility without accompanying pain.

Example B

Capsule for oral administration in accordance with the present invention has the following composition, %/mm:

Connection example 3 - 15

The glycol - 85

Other compounds having a structure corresponding to formula I, is used essentially with the same results.

A man weighing 90 kg (198 lb) suffering from osteoarthritis, treated by the method according to the present invention. In particular, he was introduced within 5 years of daily capsule containing 70 mg of the compound of example 3.

At the end of the treatment period the patient was investigated using rotoscope and found that the progression of erosion and fibrillation of the articular cartilage is missing.

Example C

Composition based salt solution for local swedeneurope alcohol - 15

Saline - 80

Other compounds having a structure corresponding to formula I is used, essentially, with the same results.

A person with severe corneal ulceration has introduced a drop of the drug in each eye twice a day. The treatment was successful without complications visible.

Example D

Composition for local introduction in accordance with the present invention, has the following composition, % m/m:

Connection example 3 - 0,20

The benzalkonium chloride - 0.02

Thimerosal - 0,002

d-Sorbitol - 5,00

Glycine - 0,35

Flavors - 0,075

Purified water For consumption.

Total: - 100,00

Any other connection structure corresponding to the formula I, is used essentially with the same results.

The patient with chemical burns inflicted composition at each dressing change (twice a day). Scarring is significantly decreased.

Example E

Aerosol composition for inhalation according to the present invention has the following composition, % m/m:

Connection example 2 - 5,0

Alcohol - 33,0

Ascorbic acid - 0,1

Menthol - 0,1

Sodium saccharin - 0,2

The gas propellant (F12, F114) requirements.

Patient with asthma use inhalers were injected himself 0.01 ml of the drug in the mouth by inhalation. Asthma symptoms were loosened.

Example F

Ophthalmic composition for local application in accordance with the present invention has the following composition, % m/m:

The compound of example 5 - 0,10

The benzalkonium chloride is 0.01

EDTA - 0,05

Hydroxyethylcellulose (NATROSOL M) - 0,50

Metabisulphite sodium - 0,10

Sodium chloride (0.9 per cent) consumption.

Total: - 100,0

Any other compounds having a structure corresponding to formula I is used, essentially, with the same results.

A man weighing 90 kg (198 pounds) with ulcerations of the cornea was treated by the method according to the present invention. In particular, in his affected eye was injected twice a day for 2 months saline solution containing 10 mg of the compound of example 5.

Example G

The composition for parenteral administration has the following composition:

Component - Number

Example 4 to 100 mg/ml carrier

Media:

Natriciteres buffer with (percent by weight of carrier): - 0,48

lecithin - 0,48

the carboxymethyl cellulose - 0,53

the povidone - 0,50

metallization 2.0 ml of the suspension is administered by injection to a person with premetastatic tumor. The injection - near the tumor. This dose is administered twice a day for about 30 days. After 30 days, the symptoms of the disease are attenuated and the dose is gradually reduced to a maintenance condition of the patient level.

Other compounds having a structure corresponding to formula I, is used essentially with the same results.

Example H

Composition for mouth, % m/about:

Connection example 1 - 3,00

Alcohol SDA 40 - 8,00

Corrigent - 0,08

Emulsifier - 0,08

Sodium fluoride - 0,05

Glycerin - 10,00

Podslushivaet - 0.02

Benzoic acid - 0,05

Sodium hydroxide - 0,20

Dye - 0.04

Water Up to 100

The patient with diseased gums uses 1 ml of fluid for rinsing the mouth three times a day to prevent further intraoral degeneration.

Other compounds having a structure corresponding to formula I is used, essentially, with the same results.

Example 1

The composition in the form of tablets, % m/about:

Connection example 3 - 0,01

Sorbitol - 17,50

Mannitol - 17,50

Starch - 13,60

Podslushivaet - 1,20

Corrigent - 11,70

Dye - 0,10

Corn syrup - 100

The patient priminary, corresponding to the formula I, is used essentially with the same results.

Example J

The composition in the form of chewing gum, % m/about:

Connection example 1 - 0,03

Crystals of sorbitol - 38,44

The basis of Paloja gum-T - 20,00

Sorbitol (70% aqueous solution) - 22,00

Mannitol - 10,00

Glycerin - 7,56

Corrigent - 1,00

The patient is chewing gum for preventing loosening of the teeth.

Other compounds having a structure corresponding to formula I is used, essentially, with the same results.

Example TO

Components, % m/about:

Water USP (pharmaceutical purity according to United States Pharmacopeia) - 54,656

Methylparaben - 0,05

Propylparaben - 0,01

Xanthan gum - 0,12

Guar gum - 0,09

Calcium carbonate - 12,38

Protivovospalitel - 1,27

Sucrose - 15,0

Sorbitol - 11,0

Glycerin - 5,0

Benzyl alcohol - 0,2

Citric acid - 0,15

A cooling substance - 0,00888

Corrigent - 0,0645

Dye - 0,0014

Composition was prepared by first mixing 80 kg of glycerol and just benzyl alcohol and heated to 65oC, then slowly add and mix methylparaben, propylparaben, in the power coaxial agitator of Silverson (Silverson). Then slowly add the following components in the following order: the remaining glycerin, sorbitol, protivovospalitel C, calcium carbonate, citric acid and sucrose. Separately combine corrigentov and dyes, and then slowly add them to other components. Mix for about 40 minutes.

The patient takes the drug to prevent a sharp aggravation of colitis.

All sources of information mentioned in this description are incorporated into it by reference.

Were described specific variants of the present invention, but for specialists in this area it is obvious that they can be made a variety of changes and additions, without going beyond the nature and scope of the invention. It is assumed that the appended claims cover all such changes and additions.

DATA ON THE BIOLOGICAL ACTIVITY

In table. 2 shows the data of in vitro tests for the biological activity of the compounds according to the invention. The test results inhibitory effect of compounds on matrix metalloproteinases (MMP enzymes) are given in respect of the following enzymes of this class:

MMR - collagenase 1

MMR - gelatinase A;

MMR - stroll the Aza 3 rats.

The test procedure is described, in particular, in the work of S. Pikul et al., J. Med.Chem., 1999, Vol. 42, No.1, pp. 87-94.

Test data are presented as values IC50that is the generally accepted measure of inhibitory activity.

1. Amides, phosphinic acids of the General formula (I)

< / BR>
where R1is hydrogen, alkyl, phenylalkyl, pyridinyl, pyridinylmethyl, alkoxyalkyl, generalkonsulat;

R2is hydrogen, alkyl, phenylalkyl, indolyl, generalkonsulat, alkylthiomethyl, acylaminoalkyl;

R3- alkyl or phenyl;

R4- alkyl, phenyl or substituted phenyl, pyridyl, thienyl or furyl,

their optical isomers, diastereomers or enantiomers, or their pharmaceutically acceptable salts, or biohydrology esters.

2. Connection on p. 1, of General formula (I)

< / BR>
where R1is hydrogen, alkyl, phenylalkyl, pyridinyl, alkoxyalkyl, generalkonsulat;

R2is hydrogen, alkyl, phenylalkyl, indolyl, generalkonsulat, alkylthiomethyl;

R3- alkyl or phenyl;

R4is phenyl, pyridyl, thienyl or furyl,

it stereoisomer or enantiomer, or its pharmaceutically acceptable salt, or biohydrology ester.

3. Soy is on PP.1-3, where R2is hydrogen or C1-C6alkyl, R4is phenyl or substituted phenyl.

5. Connection PP.1-4, where R4is phenyl, substituted alkoxy.

6. Connection PP.1-5, where R3- C1-C6lower alkyl or phenyl, R2is hydrogen, isobutyl or methyl.

7. Connection PP.1-6, where R1selected from phenyl, phenylethyl, benzyl, pyridinylmethyl or methyl.

8. The compound according to any one of the preceding paragraphs, which is selected from N - hydroxy-2-[[diphenylphosphine] (2-phenylethyl)amino] - ndimethylacetamide; N - hydroxy-2-[[methylphenylphosphinic] (2-phenylethyl)amino] - ndimethylacetamide; N - hydroxy-2(R)-[[(R)-methylphenylphosphinic] benzylamino] -4-methylpentanoate; N - hydroxy-2(R) - [[dimethylphosphine]benzylamino]-4-methylpentanoate; N - hydroxy-2(R)-[[(R)-methylphenylphosphinic]benzylamino]-propionamide; N - hydroxy-2(R) - [[diphenylphosphine] benzylamino] propionamide; N - hydroxy-2(R)-[[(R/S)-methylphenylphosphinic] -3-picolylamine] -4-methylpentanoate; N - hydroxy-2(R)-[[(S)-methylphenylphosphinic] -amino] -4-methyl-pentanolide; N - hydroxy-2(R)-[[(R)-ethylvanillin] amino] propionamide or N - hydroxy-2(R)-[[(R)-methylphenylphosphinic]-hexylamino]-propionamide.

9. The compound according to any one of paragraphs.1-8, applicable for Polo undesirable activity metalloprotease the mammal, includes introduction to the mammal a safe and effective amount of a compound according to any one of the preceding paragraphs.

 

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