MXPA01003423A - Use of catechol derivatives as proteinase inhibitors - Google Patents

Abstract

The invention relates to the use of catechol derivatives of general formula (I), wherein R1 to R5 have the meanings according to Claim No. 1, as proteinase inhibitors.

Description

USE OF CATECOL DERIVATIVES AS PROTEASE INHIBITORS DESCRIPTION OF THE INVENTION The invention relates to the use of catechol derivatives selected as protease inhibitors and their use in the treatment of diseases in which the pathogenesis of elastase or metalloproteases is involved. The metalloproteases and the serine protease elastase play a central role in the formation and progress of inflammatory diseases in the human body such as rheumatoid arthritis, periodontitis, the reaction of the skin to UV radiation, as well as mainly non-inflammatory diseases such as the formation of arteriosclerotic plaques, the mobilization of tumor cells and the formation of metastasis, osteoporosis and osteoarthritis. The role of metalloproteases in the development of processes consists, on the one hand, in the decomposition of matrix tissue and, on the other hand, in the activation of proinflammatory precursor proteins. The known inhibitors of proteases are generally complex molecules which are used for the inhibition of metalloproteases. They include, in addition to the tetracycline derivatives, for most peptidomimetics (Beckett et al., Drug Discovery Today, (1996), pp. 16-26). This gives rise to the problem of limited oral availability for peptidomimetics, since these substances are digested by relatively nonspecific peptidases in the gastrointestinal tract. Compounds of the general formula I are known, for example, from EP-A-0 202 529 and WO 96/31206 as inhibitors of lipoxygenase and antihistamines, and from WO 82/03729 as adjuvants for electrode modification. The objective of the present invention is to make available compounds which are used as protease inhibitors for the treatment of diseases in which the pathogenesis of metalloproteases or elastase is involved. It has now been found that the established requirements of the compounds are met by the selected catechol derivatives of the general formula I. The compounds differ by pronounced inhibition of the metalloprotease or elastase activity. Accordingly, the invention provides compounds selected from catechol of the general formula I wherein R 1 represents H, aryl-heterocyclyl, alkyl of 1 to 16 carbon atoms, aryl, CHO, CON (CH 3) 2, COCH 3, CO-tert-butyl, heterocyclyl or alkenyl of 2 to 16 carbon atoms; R 2 represents H, aryl-heterocyclyl, alkyl of 1 to 16 carbon atoms, aryl, CHO, CON (CH 3) 2, COCH 3, CO-tert butyl, heterocyclyl or alkenyl of 2 to 16 carbon atoms; RJ represents H, OH, alkoxy of 2 to 16 carbon atoms or alkenyl (of 2 to 6 carbon atoms) -COO-alkyl of 1 to 6 carbon atoms; R4 represents aryl, -CH = CH-aryl, H, mono- or di-alkynyl of 2 to 16 carbon atoms unsubstituted or substituted by OH-, NH2- or halo, heterocyclyl, -C (O) -heterocyclyl, alkyl from 1 to 8 carbon atoms, or R3 and R4, together with the carbon atoms of the aromatic ring, form a saturated or partially unsaturated cyclic hydrocarbon ring, of 4 to 6 members; And R5 represents aryl, -CH = CH-aryl, H, mono- or di-alkynyl of 2 to 16 carbon atoms unsubstituted or substituted with OH-NH, halo, heterocyclyl, • C (O) heterocyclyl, alkyl of 1 at 8 carbon atoms, or R4 and R5, together with the carbon atoms of the aromatic ring, form a saturated or partially unsaturated cyclic hydrocarbon ring, of 4 to 6 members; in the form of bases of pharmaceutically acceptable salts which are used as protease inhibitors in the treatment of diseases in which the pathogenesis of elastase or metalloproteases is involved. In the present invention, the expression "alkyl of 1 to 16 carbon atoms" means straight or branched chain hydrocarbons having 1 to 16 carbon atoms. Mention may be made, by way of example, of methyl, ethyl, propyl, n-butyl, sec-butyl, n-pentyl, neopentyl, n-hexyl, n-heptyl, 2,3,4-trimethylheptyl, 2, 2, 3, 4, 5, 5, 6-heptamethyloctyl, n-nonyl, 2,6 -diethyl-3, 4, 5-trimethyldecanoyl, n-undecanyl, n-dodecanyl and 3-ethyl-4-methyldodecanyl. Within the scope of the present invention, the term "C 1-16 alkoxy" means straight or branched chain hydrocarbons having 1 to 16 carbon atoms, as defined above, which are attached via the carbon atom. oxygen . The terms "alkenyl of 2 to 6 carbon atoms" and "alkenyl of 2 to 16 carbon atoms" in the present invention mean straight or branched chain hydrocarbons having 2 to 6 or 2 to 16 carbon atoms, respectively, as defined above, which additionally contain a free double bond within the carbon chain.

Within the scope of the present invention "mono- or di-alkynyl of 2 to 16 carbon atoms" means straight or branched chain hydrocarbons having from 2 to 16 carbon atoms, as defined above, which additionally contain one or two triple free links within the carbon chain. The hydrocarbon radical can additionally contain substituents of the OH, NH2 or halogen group, or any of them. The term "aryl" within the scope of the present invention means phenyls or naphthyl which are unsubstituted or monosubstituted or polysubstituted by OH, F, Cl, CF3, alkyl of 1 to 16 carbon atoms, alkoxy of 1 to 16 carbon atoms. carbon, cycloalkyl of 3 to 7 carbon atoms, alkenyl of 2 to 16 carbon atoms, heterocyclyl or of phenyl. The heterocyclyl or phenyl radicals may optionally be fused. Within the scope of the present invention, the term "heterocyclyl" means 5- or 6-membered saturated or unsaturated heterocyclic compounds which contain one or two heteroatoms of the nitrogen, oxygen or sulfur group, which optionally are provided with a condensed system in the aryl and which are unsubstituted or monosubstituted or polysubstituted by OH, F, Cl, CF3, alkyl of 1 to 16 carbon atoms, alkoxy of 1 to 16 carbon atoms, alkenyl of 2 to 16 carbon atoms, mono- or dialkynyl from 2 to 16 carbon atoms, saturated or unsaturated carboxylic acids or carboxylic acid esters of 2 to 16 carbon atoms having from 2 to 16 carbon atoms for the hydrocarbon part of the carboxylic acid and from 1 to 6 carbon atoms for the hydrocarbon part in the ester, for heterocyclyl or for phenyl. Examples of saturated heterocyclyls which may be mentioned are 1,4-dioxane, tetrahydrofuran, pyrrolidine, oxazolidine and 1,4-thioxane. From the group of the unsaturated heterocycles can be mentioned, by way of example, furan, thiophene, pyridine, pyrimidine, pyrazole, thiopyran, pyran, thiazole, oxazole, isoxazole, pyridazine, pyrazine, quinoline, isoquinoline, phthalazine and quinazoline. Within the scope of the present invention, the term "aryl-heterocyclyl" is to be understood as meaning "aryl" and "heterocyclyl" as defined above, which are joined to each other by means of a single bond. Preference is given to compounds in which the radicals R1 and R2 represent hydrogen and the radicals R3 and R5 are as defined above. Preference is given to compounds which are selected from the group: 5,6,7-trihydroxy-3,4-dihydronaphthalene-2-carboxylic acid methyl ester; 4-nonyloxy-l, 2-benzenediol; (3, -dihydroxy-phenyl) (5-phenyl-2H-pyrazol-3-yl) -methanone; 4- (2-naphth-2-yl) vinyl) -1,2-benzenediol; 3 - (6-Hept-1-ynyl-2,3-dihydroxyphenyl) -aryl methyl ester; and 4- (11-hydroxy-undeca-1,9-diinyl) -1,2-benzenediol. The compounds of the general formula I according to the invention are used in the treatment of diseases of the group of rheumatoid arthritis, periodontitis, arteriosclerotic plaques, osteoporosis, arthrosis, formation of metastases and neoangiogenesis of tumors, corneal ulceration and reactions of the skin to UV radiation. The compounds of the general formula I are used in particular in the treatment of rheumatoid arthritis, parodontitis, atherosclerotic plaques, osteoporosis, osteoarthritis and the formation of metastases and tumor neoangiogenesis. The catechol derivatives of the general formula I are preferably used in the treatment of osteoporosis, arteriosclerotic plaques and osteoarthritis.

Eg emplos The following examples serve to demonstrate the activity according to the invention of the catechol compounds of the general formula I in a matrix of metalloproteases (MMP) and elastase. For this purpose, experiments are carried out using various matrix metalloproteases from the group of human fibroblast collagenase (MMP-1), stromelysin-1 (MMP-3), human leukocyte collagenase (MMP-8) and human gelatinase B ( MMP-9) and on human elastase, which showed that the inhibition is carried out by the protease inhibitors according to the invention.

In vi tro experiments were carried out using purified human enzymes. The substrates of the enzyme used were substrates that were measured by photometry or fluorometry. The results are summarized in Table 1 below. The data demonstrate that the compounds of the formula generates 1 have metalloprotease inhibitory properties. Elastase inhibition has also been demonstrated. The compounds show variable activity in the inhibition of elastase. The following compounds were used in the examples below in the specified sequence: Example 1: 5,6,7-trihydroxy-3,4-dihydronaphthalene-2-carboxylic acid methyl ester; Example 2: 3- (6-Hept-l-inyl-2,3-dihydroxyphenyl) -aryl methyl ester; Example 3: 4-nonyloxy-l, 2-benzenediol. Example 4: 4- (11-hydroxy-undeca-1,9-diinyl) -1,2-benzenediol; Example 5: 4- (2-naphth-2-yl-vinyl) -1,2-benzenediol; Example 6: (3,4-dihydroxy-phenyl) (5-phenyl-2H-pyrazol-3-yl) methanone.

Table 1: Action of metalloproteases and elastases in vi tro I I As well as the inhibitory metalloproteases, the compounds of the general formula I also show inhibition with respect to the elastase (table 1). In an additional experiment in vi tro the effectiveness of the compounds is tested when considering inflammatory diseases in an organoid model of cartilage formation. For this model, mesenchymal cells from mouse embryo member nuclei are placed in a cell culture. In cell culture, the cells form an extracellular matrix that corresponds to the cartilage of the joint. Mineralization of the extracellular matrix is induced by the addition of β-glycerophosphate. The formation of cartilage and mineralization are inhibited by the addition of bacterial lipopolysaccharide (LPS) as an inflammation mediator. The effect of the compounds on the LPS-induced damage of the mineralization is examined by measuring the calcium content of the extracellular matrix by flame photometry after cultivation for 14 days. The effect of cartilage formation is demonstrated by checking the proteoglycan content of the extracellular matrix using Alcian blue as a dye after a treatment period of 4 to 16 days.

Table 2: Effect of compounds that have in vitro activity in the organoid model The values indicate the percentage increase in mineralization compared to a culture treated with LPS. The results shown in Table 2 demonstrate that the reductions induced by LPS in the matrix mineralization are antagonized by means of the compounds according to the invention. An increased binding of Alcian blue for the proteoglycan content of the extracellular matrix serves as a measure of the condoprotective action. The compounds stimulate the preservation of the extracellular matrix not only in the presence but also in the absence of LPS. The increase in the binding of Alcian blue depends on the concentration. Some compounds show over the course of time that they induce a massive condoprotective effect by inhibiting decomposition. This effect is accompanied by a marked inhibition of the release of prostaglandin E2 (PGE2) in the culture medium of the organoid model cells. It has also been found that increased concentrations of PGE2 in the synovial fluid of arthritic patients. Prostaglandin E2 (PGE2) is a known product of the metabolism of arachidonic acid from the cyclooxygenase pathway. This prostaglandin increases the pronociceptive action of bradykinin, causes vasodilation and has an activity that promotes edema. In an additional test, the surprising results of the in vi tro example and the findings of the murine organoid model are verified using the example of rheumatoid arthritis. The catechol derivatives are tested in a cell culture of human synovial fibroblasts. The cellular material used for the experiments originates from arthritic knee joints. Fibroblasts cultures are treated with various lipopolysaccharide (LPS) stimuli, interleukin-1 (IL-1) and a neuropeptide, substance P. Lipopolysaccharide and interleukin-1 are proinflammatory mediators. Substance P is a neurotransmitter of afferent neurons which is involved in inflammatory processes and pprocessing (N. Otsuka and K. Yoshioka, The Am. Physiol. Soc., Vol. 73, No. 2, April 1993 , 229-308). The release of interleukin-6 (IL-6), a mediator of inflammation, and the activity of secreted metalloproteases are used as measurement parameters. The release of IL-6 is inhibited by catechol derivatives according to the invention. The matrix metalloprotease activity in the cell culture medium is measured using a fluorogenic substance (Knight CG et al. (1992) FEBS Lett 296, 263). The proteolytic activity of the samples without adhesion of catechol derivatives of the general formula I is considered to be 100%. The cell culture medium treated with the compounds at a concentration of (10 ~ 5 M) shows inhibition of proteolytic activity independently of in vitro activation of proenzymes. After incubation with trypsin and trypsin inhibitor (in this system, interstitial collagenase MMP-1 is mainly activated), 90% inhibition is observed in the case of some compounds, while in the case of activation by 4-acetate. aminophenylmercury (APMA) which activates mainly gelatinase A, initiates only a slight inhibition. The activity of stromelysin-1 (MMP-3), measured using a specific fluorogenic substrate (Nagase et al. (1994), J. Biol. Chem. 269, 20952), is inhibited by 85%, compared to the untreated control medium.

The stimulation of synovial fibroblasts by lipopolysaccharides, IL-1 and the neuropeptide substance P leads, in the presence of catechol derivatives according to the invention, to a reduced activity of the matrix metalloprotease. After stimulation by lipopolysaccharide, the activity of matrix metalloprotease (MMP) activatable by 4-aminophenylmercury acetate (APMA), that is, gelatinase A in this case, is inhibited only slightly, while matrix metalloprotease activity activatable by trypsin (MMP), that is, stromelysin-1 (MMP-3) is inhibited. The inhibitory action of the compounds in the metalloprotease activities of synovial fibroblast matrix stimulated with IL-1 in the same way can be demonstrated. Matrix metalloprotease activity activable by APMA, ie, gelatinase A, is not inhibited after stimulation by the neuropeptide of substance P, whereas matrix metalloproteinases activatable by trypsin, ie, mainly interstitial collagenase and stromelysin-1, are inhibited by 75% .

Claims (6)

1. Use of catechol derivatives of the general formula I wherein R 1 represents H, aryl-heterocyclyl, alkyl of 1 to 16 carbon atoms, aryl, CHO, CON (CH 3) 2, COCH 3, CO-tert-butyl, heterocyclyl or alkenyl of 2 to 16 carbon atoms; R 2 represents H, aryl-heterocyclyl, alkyl of 1 to 16 carbon atoms, aryl, CHO, CON (CH 3) 2, COCH 3, CO-tert-butyl, heterocyclyl or alkenyl of 2 to 16 carbon atoms; R3 represents H, OH, alkoxy of 2 to 16 carbon atoms or alkenyl (of 2 to 6 carbon atoms) -COO-alkyl of 1 to 6 carbon atoms; R 4 represents aryl, -CH = CH-aryl, H, mono- or di-alkynyl of 2 to 16 carbon atoms unsubstituted or substituted by OH-NH, halo, heterocyclyl, -C (O) heterocyclyl, alkyl of 1 to 8 carbon atoms, or R3 and R4, together with the carbon atoms of the aromatic ring, form a saturated or partially unsaturated cyclic hydrocarbon ring, of 4 to 6 members; and R5 represents aryl, -CH = CH-aryl, H, mono- or di-alkynyl of 2 to 16 carbon atoms unsubstituted or substituted with -OH-, NH2- or halo, heterocyclyl, -C (O) -heterocyclyl , alkyl of 1 to 8 carbon atoms, or R4 and R5, together with the carbon atoms of the aromatic ring, form a saturated or partially unsaturated cyclic hydrocarbon ring, of 4 to 6 members; in the form of pharmaceutically acceptable salts bases, as protease inhibitors in the treatment of diseases in which the pathogenesis of elastase or metalloproteases is involved.

2. The use of catechol derivatives, as described in claim 1, wherein R1 and R2 represent hydrogen and the radicals R3 to Re are as defined according to claim 1.

3. The use of catechol derivatives, as described in claim 1, which are selected from the group: 5,6,7-trihydroxy-3,4-dihydronaphthalene-2-carboxylic acid methyl ester, 4-nonyloxy-2, -benzenediol; (3,4-dihydroxy-phenyl) (5-phenyl-2H-pyrazol-3-yl) -methanone; 4- (2-naphth-2-yl) vinyl) -1,2-benzenediol; 3- (6-Hept-l-inyl-2,3-dihydroxyphenyl) -aryl acid methyl ester; and 4- (11-hydroxy-undeca-1,9-diinyl) -1,2-benzenediol.

4. The use of catechol derivatives, as described in claim 1, in the treatment of diseases of the group of rheumatoid arthritis, parodontitis, atherosclerotic plaques, osteoporosis, arthrosis, formation of metastases and neoangiogenesis of tumors, ulceration of the cornea and reactions of the skin to UV radiation.

5. The use of catechol derivatives, as described in claim 1, in the treatment of diseases of the group of rheumatoid arthritis, parodontitis, arteriosclerotic plaques, osteoporosis, osteoarthritis, metastasis formation and neoangiogenesis of tumors.

6. The use of 4- (2-naphth-2-yl-vinyl) -1,2-benzenediol as described in claim 1, in the treatment of diseases of the group of rheumatoid arthritis, parodontitis, osteoporosis, arteriosclerotic plaques, osteoarthritis, formation of metastasis and neoangiogenesis of tumors.