WO2002020566A2 - NOVEL MMP-2 DERIVATIVES FOR USE AS INHIBITORS OF INTEGRIN αVβ¿3? - Google Patents
NOVEL MMP-2 DERIVATIVES FOR USE AS INHIBITORS OF INTEGRIN αVβ¿3? Download PDFInfo
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- WO2002020566A2 WO2002020566A2 PCT/EP2001/009899 EP0109899W WO0220566A2 WO 2002020566 A2 WO2002020566 A2 WO 2002020566A2 EP 0109899 W EP0109899 W EP 0109899W WO 0220566 A2 WO0220566 A2 WO 0220566A2
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- C12N9/6489—Metalloendopeptidases (3.4.24)
- C12N9/6491—Matrix metalloproteases [MMP's], e.g. interstitial collagenase (3.4.24.7); Stromelysins (3.4.24.17; 3.2.1.22); Matrilysin (3.4.24.23)
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Definitions
- the invention relates to novel compounds of the formula I.
- Amino acids A alkyl with 1-10 C atoms
- the primary amino groups can also be provided with conventional amino protecting groups, and their physiologically acceptable salts and solvates.
- the invention was based on the task of finding new compounds with valuable properties, in particular those which can be used for the production of medicaments.
- the compounds of the formula I and their salts have very valuable pharmacological properties with good tolerability. Above all, they act as integrin inhibitors, in particular the interactions of the ⁇ y-integrin receptors with ligands, e.g.
- the compounds according to the invention are ligands in particular of
- Integrins ⁇ ß 3 which do not contain the sequence RGD and were derived from the C-terminal fragment PEX of the matrix metalloproteinase 2 (MMP-2).
- MMP-2 is a matrix-degrading enzyme that facilitates cell migration in invasive processes, e.g. in tumor metastasis and angiogenesis.
- MMPs are secreted as inactive proenzymes ("latent enzyme", “zymogen”), so enzymatically inactive Pro-MMP-2 is also known:
- MMPs allow tumor cell invasion and metastasis through at least 3 mechanisms. They remove the physical barrier by breaking down the ECM, they modulate cell adhesion since the cells have to adhere to the ECM but have to break bonds to the ECM again, and they produce other biologically active protein fragments by breaking down proteins.
- invadopodia and lamellipodia on the migration front, which mediate contact with the underlying extracellular matrix (ECM), have different expression and localization of MT1-MMP, MMP-2, and TIMP-2.
- ECM extracellular matrix
- Pro-MMP-2 and MT1-MMP are responsible for the dismantling of the ECM, since MMP activation takes place here.
- lamellipodia have a similar structure, there is no ECM degradation here, since they are easily accessible to circulating inhibitors such as TIMP-2 on the cellular periphery. It was developed by Chen-WT et al. in Ann-N-Y-Acad-Sci. 1999 Jun 30; 878: 361-71 showed that TIMP-2 with MMP-2 and MT1-MMP is colocalized on lamellipodia but not on Invadopodia. The authors Nakahara-H; et al. proc
- MMP-2 occurs in normal and diseased human joints:
- Carcinoma was developed by Davidson-B et al. in Gynecol-Oncol. 1999 Jun; 73 (3):
- MMP-2 and its activation play a role in neointima formation in restenosis: Wang-H; Keizer-JA J-Vasc-Res. 1998 Jan-Feb; 35 (1): 45-
- MMP-2 modulates glioma adhesion and migration: Deryugina-El; et al. J Cell Sci. 1997 Oct; 110 (Pt 19): 2473-82.
- the compounds are particularly effective in the case of the integrins' ⁇ vß 3 and vßs-
- the compounds are particularly effective as adhesion receptor antagonists for the receptor ⁇ vß ß .
- This effect can be demonstrated, for example, by the method described by B. Diefenbach et al. in Biology of Vitronectins and Their Receptors, Eds. KT Preissner et al., Exerpta Medica, Interntl. Congress Series 1042, Elsevier Sei. Publishers 1993, p. 149-156 is described.
- the compounds of the formula I according to the invention can therefore be used as active pharmaceutical ingredients, in particular for the treatment of tumor diseases, osteoporoses and other osteolytic diseases and for the suppression of angiogenesis.
- micro-aggregates microthrombi
- the spread of tumor cells from a local tumor into the vascular system takes place through the formation of micro-aggregates (microthrombi) through the interaction of the tumor cells with platelets.
- the tumor cells are shielded by the protection in the micro-aggregate and are not recognized by the cells of the immune system.
- the micro-aggregates can attach themselves to the vessel walls, which facilitates further penetration of tumor cells into the tissue. Since the formation of the microthrombi is mediated by fibrinogen binding to the fibrinogen receptors on activated platelets, the GPIIb / llla antagonists are considered to be effective metastasis inhibitors.
- Platelets also bind other adhesive proteins, such as vitonectin, collagen and laminin, to the corresponding receptors on the surface of various cell types. In particular, they prevent the formation of platelet thrombi and can therefore be used to treat thrombosis, apoplexy, heart attack, inflammation and arteriosclerosis.
- adhesive proteins such as vitonectin, collagen and laminin
- the properties of the compounds can also be demonstrated by methods which are described in EP-A1-0462 960.
- the inhibition of fibrinogen binding to the fibrinogen receptor can be demonstrated by the method specified in EP-A1-0 381 033.
- the antiplatelet effect can be demonstrated in vitro by the method of Born (Nature 4832, 927-929, 1962).
- the bone resorption can be inhibited by the compounds according to the invention with the aid of an osteoclast absorption test analogous to WO 95/32710.
- Matrix metalloproteinase 2 is a member of the zinc-dependent endoprotease family, which are secreted as pro-enzymes, latent MMPs or zymogens and for the degradation of extracellular matrix proteins and other proteins are responsible to enable cell migration and invasion.
- the MMPs play a role in physiological as well as pathological situations, in embryonic development, in inflammation, wound healing, tumor cell invasion and metastasis. Inhibitors of the enzymatic
- the enzyme cleaves various types of collagen, elastin, fibronectin and laminin. Binding to collagen takes place by means of 3 repeat units homologous to fibronectin type II modules, which are inserted into the catalytic domain near the active site.
- CTD C-Terminal Domain
- PEX Cheresh et al.
- Goldberg has described the amino acid sequence of Pro-MMP-2 (72 kDa) in US Pat. No. 4,923,818; the complex with TIMP-2 is documented by Goldberg in EP 404,750.
- Soluble Pro-MMP-2 is bound to cell surfaces and is there by the membrane-bound enzyme MT1-MMP by proteolysis of the ProFragmentes in position Asn66-Leu67 in the first step and subsequent MT1-MMP-independent digestion at position Asn109-Tyr110 activated. This activation is not possible if the C-terminus of Pro-MMP-2 is missing. Possible binding partners are MT1-MMP, integrin ⁇ v ß3 and Fetuin.
- WO 98/12309 Washington University St. Louis, describes binding sequences from CTD (PEX) to TIMP-2. It is mentioned that the addition of an excess of TIMP-2 (but not TIMP-1) or recombinant CTD (PEX) suppresses the activation of Pro-MMP-2 on the cell surface.
- the invention accordingly relates to compounds of the formula I according to claim 1 and / or their physiologically acceptable salts for the preparation of a medicament for use as integrin inhibitors.
- the invention relates in particular to compounds of the formula I according to Claim 1 and / or their harmless salts for the production of a medicament for combating pathologically angiogenic diseases, tumors, osteoporosis, inflammation and infections.
- the compounds of formula I can be used as active pharmaceutical ingredients in human and veterinary medicine, for the prophylaxis and / or therapy of thrombosis, myocardial infarction, arteriosclerosis, inflammation, apoplexy, angina pectoris, tumor diseases, osteolytic diseases such as osteoporosis, hypercalcaemia, pathologically angiogenic diseases such as B.
- inflammation ophthalmic diseases, diabetic retinopathy, macular degeneration, myopia, ocular histoplasmosis, rheumatoid arthritis, osteoarthritis, rubeotic glaucoma, ulcerative colitis, Crohn's disease, atherosclerosis, psoriasis, restenosis after angioplasty, infection, with infection, viral infection acute kidney failure and wound healing to support the healing processes.
- the compounds of formula I can be used as antimicrobial substances in operations where biomaterials, implants, catheters or pacemakers are used. They have an antiseptic effect.
- the effectiveness of the antimicrobial activity can be by P.Valentin-Weigund et al., in Infection and Immunity, 2851-2855 (1988).
- the invention also relates to the hydrates and solvates, e.g. Alcoholates, these compounds.
- the invention furthermore relates to a process for the preparation of compounds of the formula I according to claim 1 and their salts, characterized in that a compound of the formula I is liberated from one of its functional derivatives by treatment with a solvolysing, reducing or hydrogenolysing agent, and / or converts a base or acid of the formula I into one of its salts.
- the compounds of the formula I can have one or more chiral centers and can therefore occur in several stereoisomeric forms. All of these forms (e.g. D and L forms) and their mixtures (e.g. the DL forms) are included in Formula I. So-called prodrug derivatives are also included in the compounds according to the invention, i. H. with z. B. alkyl or acyl groups, sugars or oligopeptides modified compounds of formula I, which are quickly cleaved in the organism to the active compounds of the invention.
- Trt trityl (triphenylmethyl).
- MMP-2 The sequence of human Pro-M P-2 (MMP-2) taken from publicly available databases (e.g. available from www.expasy.ch) is as follows:
- the C-terminal, hemopexin-like region also known in the literature as PEX, from position 466-660 is marked dark.
- the integrin ⁇ v ⁇ 3 binding peptide sequences according to the invention are underlined.
- A is alkyl and has 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms and is preferably methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl or tert.
- Butyl also for pentyl, 1-, 2- or 3-methylbutyl, 1,1-, 1, 2- or 2,2-dimethylpropyl, 1-ethylpropyl, hexyl, 1-, 2-, 3- or 4 - methylpentyl, 1, 1-, 1, 2-, 1, 3-, 2,2-, 2,3- or 3,3-dimethylbutyl, 1- or
- 2-ethylbutyl 1-ethyl-1-methylpropyl, 1-ethyl-2-methylpropyl, 1, 1, 2-, 1, 2,2-trimethylpropyl, heptyl, octyl, nonyl or decyl, undecyl.
- A also means alkyl substituted by halogen, preferably CF 3 .
- Acyl means alkanoyl with 1-10 C atoms, preferably e.g. Formyl, acetyl, propionyl or butyryl, further e.g. Benzoyl.
- Amino protecting group preferably means formyl, acetyl, propionyl, butyryl, phenylacetyl, benzoyl, toluyl, POA, methoxycarbonyl, ethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, BOG, 2-iodoethoxycarbonyl, CBZ ("carbobenzoxy"), 4-methoxybenzyloxycarbonyl, FMOC, Mtr or benzyl.
- the invention relates in particular to those compounds of the formula I in which at least one of the radicals mentioned has one of the preferred meanings indicated above.
- Some preferred groups of compounds can be expressed by the following sub-formulas la to le, which correspond to the formula I and in which the radicals not specified have the meaning given for the formula I, but in which
- Amino acids selected from the peptide sequence of human Pro-MMP-2;
- Amino acids selected from the peptide sequence of human Pro-MMP-2,
- X H acyl or a peptide fragment with 1 to 20 amino acids, selected from the peptide sequence of human Pro-MMP-2,
- Y is a peptide fragment selected from the sequence region 489-497 and / or 570-585 and / or 588-597 of human Pro-MMP-2,
- Amino acids selected from the peptide sequence of human Pro-MMP-2, Y a peptide fragment selected from the sequence range 489-497 and / or 570-585 and / or 588 597 of human Pro-MMP-2,
- Z is OH, NH 2 , NHA, NA 2 or Cys
- the primary amino groups can also be provided with conventional amino protecting groups, and their physiologically acceptable salts and solvates.
- the peptides of the C-terminal sequence of MMP-2 were synthesized as fragments of 8 with 2 amino acid overlaps on paper in an Fmoc strategy.
- the quality of the synthesis was assessed using an incident light / fluorescence reader from SLT Tecan and the quantity of the Fmoc signal was evaluated after coupling steps (much fluorescence) and after splitting steps (no fluorescence).
- These peptide-loaded papers were incubated with fluorescence-labeled integrin ⁇ v ⁇ 3 , washed and evaluated with an incident light fluorescence reader from SLT-Tecan.
- the peptide sequences of the positive “SPOTs” were prepared conventionally on the resin in separate syntheses and thereby labeled with a C-terminal cysteine.
- the purified, isolated cysteine peptides were tested in a covalent binding test for binding of biotinylated integrin ⁇ v ⁇ 3 using an anti-biotin / phosphatase Reaction checked, the v ß 3 -binding, RGD-independent activity was confirmed.Of the active compounds, 2 peptides are in a consecutive sequence which is derived from the known X-ray structure of the MMP-2 C-terminus the surface of PEX.
- the compounds according to the invention prepared by way of example were produced in accordance with the prior art in a Milligen 9050 automatic synthesizer of polystyrene.
- the process can be understood by a specialist on the basis of textbook knowledge, such as in the appendix to the Novabiochem To find catalog.
- Chlorine-trityl (o-CI) anchors were used.
- Fmoc-protected amino acids with acid-labile side protection were coupled.
- the side-protected derivatives Fmoc-Ser (But), Thr (But),
- the starting materials can also be formed in situ, so that they are not isolated from the reaction mixture, but instead are immediately reacted further to give the compounds of the formula I.
- Compounds of formula I can preferably be obtained by liberating compounds of formula I from one of their functional derivatives by treatment with a solvolysing or hydrogenolysing agent.
- Preferred starting materials for solvolysis or hydrogenolysis are those which otherwise correspond to the formula I, but instead of one or more free amino and / or hydroxyl groups contain corresponding protected amino and / or hydroxyl groups, preferably those which instead of an H atom, which is connected to an N atom carry an amino protective group, in particular those which carry an R'-N group instead of an HN group, in which R 'represents an amino protective group, and / or those which have one instead of the H atom hydroxy carry a hydroxyl protective group, for example those which correspond to the formula I, but instead of a group -COOH carry a group -COOR "in which R" denotes a hydroxyl protective group.
- amino protecting group is generally known and refers to groups which are suitable for protecting (blocking) an amino group from chemical reactions, but which are easily removable after the desired chemical reaction has been carried out at other locations in the molecule. Unsubstituted or substituted acyl, aryl, aralkoxymethyl or aralkyl groups are particularly typical of such groups. Since the amino protective groups are removed after the desired reaction (or reaction sequence), their type and size is otherwise not critical; however, those with 1-20, in particular 1-8, carbon atoms are preferred.
- acyl group is to be understood in the broadest sense in connection with the present process.
- acyl groups derived from aliphatic, araliphatic, aromatic or heterocyclic carboxylic acids or sulfonic acids and, in particular, alkoxycarbonyl, aryloxycarbonyl and especially aralkoxycarbonyl groups.
- acyl groups are formyl or alkanoyl such as acetyl, propionyl, butyryl; Aralkanoyl such as phenylacetyl; Aroyl such as benzoyl or toluyl; Aryloxyalkanoyl such as POA; Alkoxycarbonyl such as methoxycarbonyl, ethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, BOG, 2-iodoethoxycarbonyl; Aralkyloxycarbonyl such as CBZ ("carbobenzoxy"), 4-methoxybenzyloxycarbonyl, FMOC; Arylsulfonyl such as Mtr.
- Preferred amino protective groups are BOC and Mtr, furthermore CBZ, Fmoc, benzyl, formyl and acetyl.
- the amino protective group is split off, depending on the protective group used, e.g. B. with strong acids, suitably with TFA or perchloric acid, but also with other strong inorganic acids such as hydrochloric acid or sulfuric acid, strong organic carboxylic acids such as Trichloroacetic acid or sulfonic acids such as benzene or p-toluenesulfonic acid.
- strong acids suitably with TFA or perchloric acid
- other strong inorganic acids such as hydrochloric acid or sulfuric acid
- strong organic carboxylic acids such as Trichloroacetic acid or sulfonic acids
- benzene or p-toluenesulfonic acid strong organic carboxylic acids
- the presence of an additional inert solvent is possible, but not always necessary.
- Suitable inert solvents are preferably organic, for example carboxylic acids such as acetic acid, ethers such as tetrahydrofuran or dioxane, amides such as DMF, halogenated hydrocarbons such as dichloromethane, and also alcohols such as methanol, ethanol or isopropanol, and water. Mixtures of the abovementioned solvents are also suitable. TFA is preferably used in excess without the addition of another solvent, perchloric acid in the form of a mixture of acetic acid and 70% perchloric acid in a ratio of 9: 1.
- the reaction temperatures for the cleavage are advantageously between about 0 and about 50 °, preferably between 15 and 30 ° (room temperature).
- the groups BOC, OBut, Pmc, Pbf and Mtr can e.g. B. preferably with TFA in dichloromethane or with about 3 to 5N HCl in dioxane at 15-30 °, the FMOC group with an about 5 to 50% solution of sec. Amines such as dimethylamine, diethylamine or piperidine in DMF at 15-30 °.
- Hydrogenolytically removable protective groups can e.g. B. by treatment with hydrogen in the presence of a catalyst (z. B. a noble metal catalyst such as palladium, advantageously on a support such as coal).
- a catalyst z. B. a noble metal catalyst such as palladium, advantageously on a support such as coal.
- Suitable solvents are the above, especially z. B. alcohols such as
- Methanol or ethanol or amides such as DMF Methanol or ethanol or amides such as DMF.
- the hydrogenolysis is generally carried out at temperatures between about 0 and 100 ° and pressures between about 1 and 200 bar, preferably at 20-30 ° and 1-10 bar.
- Hydrogenolysis of the CBZ group succeeds e.g. B. good on 5 to 10% Pd / C in methanol or with ammonium formate (instead of hydrogen) on Pd / C in methanol / DMF at 20-30 °.
- Suitable inert solvents are, for example, hydrocarbons such as hexane, petroleum ether, benzene, toluene or xylene; chlorinated hydrocarbons such as trichlorethylene, 1, 2-dichloroethane, carbon tetrachloride, chloroform or dichloromethane; Alcohols such as methanol, ethanol, isopropanol, n-propanol, n-butanol or tert-butanol; Ethers such as diethyl ether, diisopropyl ether, tetrahydrofuran (THF) or dioxane; Glycol ethers such as ethylene glycol monomethyl or monoethyl ether (methyl glycol or ethyl glycol), ethylene glycol dimethyl ether (diglyme); Ketones such as acetone or butanone; Amides such as acetamide, dimethylacetamide or dimethylformamide (DMF); Ni
- estonify an ester of the formula I is conveniently done by solvolysis or hydrogenolysis, as indicated above, e.g. with LioH in methanol, NaOH or KOH in dioxane water at temperatures between 0 and 60 ° C, preferably between 10 and 40 ° C.
- a base of the formula I can be converted into the associated acid addition salt using an acid, for example by reacting equivalent amounts of the base and the acid in an inert solvent such as ethanol and subsequent evaporation.
- an inert solvent such as ethanol and subsequent evaporation.
- acids that provide physiologically acceptable salts are suitable for this implementation.
- inorganic acids can be used, for example sulfuric acid, nitric acid, hydrohalic acids such as hydrochloric acid or hydrobromic acid, phosphoric acids such as orthophosphoric acid, sulfamic acid, and also organic acids, in particular aliphatic, alicyclic, aromatic, heterocyclic, mono- or polycarbonate, sulfone - or sulfuric acids, for example formic acid, acetic acid, propionic acid, pivalic acid, diethyl acetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, lactic acid, tartaric acid, malic acid, citric acid, gluconic acid, Ascorbic acid, nicotinic acid, isonicotinic acid, methane or ethanesulfonic acid, ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic
- an acid of the formula I can be converted into one of its physiologically acceptable metal or ammonium salts by reaction with a base.
- Suitable salts are in particular the sodium, potassium, magnesium, calcium and ammonium salts, and also substituted ammonium salts, e.g. B. the dimethyl, diethyl or diisopropyl ammonium salts, monoethanol, diethanol or diisopropylammonium salts, cyclohexyl, dicyclohexylammonium salts, di- [benzylethylenediammonium salts, z. B. salts with arginine or lysine.
- the compounds of the formula I contain one or more chiral centers and can therefore be present in racemic or in optically active form. Racemates obtained can be separated mechanically or chemically into the enantiomers by methods known per se. Diastereomers are preferably formed from the racemic mixture by reaction with an optically active release agent. Suitable release agents are e.g. optically active acids, such as the D and L forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid or the various optically active camphorsulfonic acids such as ß-camphorsulfonic acid. Enantiomer separation using a column filled with an optically active separating agent (e.g.
- a suitable solvent is e.g. a mixture of hexane / isopropanol / acetonitrile, e.g. in the volume ratio 82: 15: 3.
- optically active compounds of the formula I by the methods described above by using starting materials which are already optically active.
- the invention includes not only the compounds mentioned, but also mixtures and preparations which, in addition to these compounds according to the invention, also include other pharmacological active ingredients or
- Contain adjuvants which can influence the primary pharmacological action of the compounds according to the invention in a desired manner. These can be used as therapeutic agents, diagnostic agents or as reagents
- Find use They can be given to humans or animals locally or systemically, orally, intravenously, intraperitoneally, intramuscularly, subcutaneously, transdermally, nasally, buccally, or iontophoretically, which includes formulations in suspensions, emulsions or solutions, liposomes, ointments, pastes, biodegradable polymers or as nanoparticles, tablets, capsules or pills, granules or powder, as an aerosol for inhalation, as intranasal drops or sprays. It is also possible to combine the new products with other technologies such as surgery, radiation, diagnosis, radiotherapy, photodynamic therapy and gene therapy, as well as with other medications.
- Such drugs can e.g. from the areas of cardiovascular,
- Tumor agents such as angiogenesis inhibitors or cytostatics
- Substances can be low molecular weight and high molecular weight. It can be
- Lipids Lipids, carbohydrates or proteins. This includes cytokines,
- Toxins fusion proteins, monoclonal antibodies and vaccines.
- the invention accordingly relates to compounds of the formulas defined above and below and in the claims, including their physiologically acceptable salts, as medicaments, diagnostic agents or reagents.
- these are drugs for combating diseases based on an interaction of ligands, for example of MMP-2 with the v ß 3
- Integrin receptor based Integrin receptor based.
- the drugs are suitable for fighting thrombosis
- Heart attack coronary heart disease, arteriosclerosis, tumors,
- Corresponding pharmaceutical preparations which contain at least one medicament of the formula I and, if appropriate, carriers and / or auxiliaries are also an object.
- the invention furthermore relates to the use of the compounds according to the invention and / or their physiologically harmless
- the invention further relates to the use of the compounds according to the invention and / or their physiologically acceptable salts and / or solvates for the manufacture of a medicament for combating diseases which are based on an interaction of a ligand, for example the matrix metalloproteinase (MMP-2) with the v ß 3 integrin receptor based. - '
- MMP-2 matrix metalloproteinase
- the invention furthermore relates to the use of the compounds according to the invention and / or their physiologically acceptable salts and / or solvates for the production of a medicament for combating pathological processes which are maintained or propagated by angiogenesis.
- the invention furthermore relates to the use of the compounds according to the invention and / or their physiologically acceptable salts and / or solvates for the manufacture of a medicament for combating thromboses, heart attacks, coronary heart diseases, arteriosclerosis, tumors, osteoporosis, fibrosis, inflammation, Infections, psoriasis and to influence wound healing processes.
- Suitable carriers are organic or inorganic substances which are suitable for enteral (e.g. oral), parenteral, topical application or for application in the form of a
- 10 inhalation sprays are suitable and do not react with the new compounds, for example water, vegetable oils, benzyl alcohols, alkylene glycols, polyethylene glycols, glycerol triacetate, gelatin, carbohydrates such as lactose or starch, magnesium stearate, talc, petroleum jelly.
- water vegetable oils, benzyl alcohols, alkylene glycols, polyethylene glycols, glycerol triacetate, gelatin, carbohydrates such as lactose or starch, magnesium stearate, talc, petroleum jelly.
- ⁇ C application are used in particular tablets, pills, dragees, capsules, powders, granules, syrups, juices or drops, for rectal application positorien Sup-, for parenteral application of solutions, preferably oily or aqueous solutions, furthermore suspensions, emulsions or implants, and for topical application ointments, creams or powder.
- solutions preferably oily or aqueous solutions, furthermore suspensions, emulsions or implants, and for topical application ointments, creams or powder.
- 20 compounds can also be lyophilized and the resulting lyophilisates e.g. can be used for the production of injectables.
- the specified preparations can be sterilized and / or auxiliaries such as lubricants, preservatives, stabilizers and / or wetting agents, emulsifiers.
- Sprays can be used for the application as an inhalation spray,
- OQ which contain the active substance either dissolved or suspended in a propellant gas or propellant gas mixture (e.g. C0 2 or chlorofluorocarbons).
- a propellant gas or propellant gas mixture e.g. C0 2 or chlorofluorocarbons.
- the active ingredient is expediently used in micronized form, with one or more additional physiologically tolerable ones
- Solvents may be present, e.g. B. ethanol. inhalation solutions
- the substances according to the invention can generally be administered in analogy to other known, commercially available preparations (for example described in US Pat. No. 4,472,305), preferably in doses between about 0.05 and 500 mg, in particular between 0.5 and 100 mg per dosage unit.
- the daily dosage is preferably between about 0.01 and 20 mg / kg body weight.
- the specific dose for each patient depends on a variety of factors, for example on the effectiveness of the particular compound used, on the age, body weight, general health, sex, on the diet, on the time and route of administration, on the rate of elimination, combination of drugs and severity the respective disease to which the therapy applies. Parenteral administration is preferred.
- HPLC analyzes (retention time Rt) were carried out in the following system:
- Trifluoroacetic acid at a flow of 1 ml / min and detection at 215 nm.
- Production and purification of peptides according to the invention In principle, production and purification is carried out by means of the Fmoc strategy while protecting acid-labile side chains on acid-labile resins using a commercially available "continuous flow" peptide synthesizer in accordance with the information provided by Haubner et al. (J. Am. Chem. Soc. 118, 1996, 17703).
- VKKKMDPGC MMP-2 (594 603) -Cys, M 1005.23;
- LERGYPKPLTSLGLC MMP-2 (548 561) -Cys, M 1646.94;
- KPMGPLLVC MMP-2 (502 • 509) -Cys, M 957.24;
- AGDKFWRYNC MMP-2 (585 • 593) -Cys, M 1259.42;
- RGYPKPLTSLC MMP-2 (550 • 559) -Cys, M 1234.5;
- PGFPKLIAC MMP-2 (600 • 607) -Cys, M 945.19;
- WRTVTPRDC MMP-2 (494 • 501) -Cys, M 1133.31;
- Placenta can be cleaned as described by Smith et al. J. Biol. Chem. 263, 18726-
- Soluble ⁇ v ß 3 is like in RJ
- Binding test is S. Kraft et al. J. Biol. Chem. 274, 1979-1985 (1999).
- the receptor binding test on covalently bound peptides 5 can be carried out analogously to: B. Diefenbach et al. in Biology of Vitronectins and Their Receptors; eds. KT Preissner et al. from Exerpta Medica International Congress Series 1042, Elsevier Science Publishers Amsterdam 1993, p. 149-156.
- fluorescein-labeled integrin ⁇ vß 3 0 was used, which interacts directly with the cellulose-bound peptide. The evaluation was carried out by measuring the fluorescence.
- BSA coated in microtiter plates was activated with sulfo-SMPB and the thiopeptides were covalently bound to it. After 5 incubations with biotinylated ⁇ vß 3 , bound receptor with alkaline-phosphatase-coupled goat anti-biotin antibody and coloring by a phosphatase substrate was detected.
- the binding of the PEX peptides to ⁇ vß 3 is an RGD-independent binding. This was verified in a further test which was carried out analogously to 1..
- the receptor was incubated with and without the soluble peptide cyclo- (Arg-Gly-Asp-D-Phe-N-Me-Val). While the binding of the cyclo-RGD peptide "cyclo- (RGDfK (mercapto-propionyl))" to ⁇ vß 3 was inhibited by cyclo- (Arg-Gly-Asp-D-Phe-N-Me-Val), the binding was of the PEX peptide AAFNWSKNKKTYIFAGC regardless of this.
- Example A Injection glasses
- a solution of 100 g of Lys-Asp-Arg-Phe-Ile-Trp-Arg-Thr-Val-Cys and 5 g of disodium hydrogen phosphate is adjusted to pH 6.5 in 3 l of double-distilled water with 2 N hydrochloric acid, sterile filtered, in Injection glasses filled, lyophilized under sterile conditions and sealed sterile. Each injection jar contains 5 mg of active ingredient.
- a mixture of 20 g of Lys-Asp-Arg-Phe-Ile-Trp-Arg-Thr-Val-Cys is melted with 100 g of soy lecithin and 1400 g of cocoa butter, poured into molds and allowed to cool.
- Each suppository contains 20 mg of active ingredient.
- a solution is prepared from 1 g of Lys-Asp-Arg-Phe-Ile-Trp-Arg-Thr-Val-Cys, 9.38 g of NaH 2 P0 4 ⁇ 2 H 2 0, 28.48 g Na 2 HP0 4 • 12 H 2 0 and 0.1 g
- Benzalkonium chloride in 940 ml of double distilled water. It is adjusted to pH 6.8, made up to 1 I and sterilized by irradiation. This solution can be used in the form of eye drops.
- Example D ointment
- Example F coated tablets
- Example E tablets are pressed, which are then coated in a conventional manner with a coating of sucrose, potato starch, talc, tragacanth and colorant.
- a solution of 1 kg of Lys-Asp-Arg-Phe-Ile-Trp-Arg-Thr-Val-Cys in 60 l of double-distilled water is sterile filtered, filled into ampoules, lyophilized under sterile conditions and sealed sterile. Each ampoule contains 10 mg of active ingredient.
Abstract
Description
Claims
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
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EP01980289A EP1315804A2 (en) | 2000-09-07 | 2001-08-28 | Novel mmp-2 derivatives for use as inhibitors of integrin alpha vbeta 3 |
AU2002212172A AU2002212172A1 (en) | 2000-09-07 | 2001-08-28 | Novel mmp-2 derivatives for use as inhibitors of integrin alphavbeta3 |
CA002421283A CA2421283A1 (en) | 2000-09-07 | 2001-08-28 | Novel mmp-2 derivatives for use as inhibitors of integrin .alpha.v.beta.3 |
JP2002525185A JP2004510710A (en) | 2000-09-07 | 2001-08-28 | Novel MMP-2 derivatives for use as inhibitors of integrin αvβ3 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
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DE10044325A DE10044325A1 (en) | 2000-09-07 | 2000-09-07 | New MMP-2 derivatives as inhibitors of the integrin avbeta3 |
DE10044325.7 | 2000-09-07 |
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WO2002020566A2 true WO2002020566A2 (en) | 2002-03-14 |
WO2002020566A3 WO2002020566A3 (en) | 2002-06-27 |
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EP (1) | EP1315804A2 (en) |
JP (1) | JP2004510710A (en) |
AU (1) | AU2002212172A1 (en) |
CA (1) | CA2421283A1 (en) |
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WO (1) | WO2002020566A2 (en) |
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US10517599B2 (en) | 2015-08-26 | 2019-12-31 | Ethicon Llc | Staple cartridge assembly comprising staple cavities for providing better staple guidance |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4923818A (en) * | 1987-09-04 | 1990-05-08 | Washington University | DNA clone of human type IV collagenase |
WO1998012309A2 (en) * | 1996-09-17 | 1998-03-26 | Washington University | Method of cell surface activation and inhibition |
-
2000
- 2000-09-07 DE DE10044325A patent/DE10044325A1/en not_active Withdrawn
-
2001
- 2001-08-28 EP EP01980289A patent/EP1315804A2/en not_active Withdrawn
- 2001-08-28 WO PCT/EP2001/009899 patent/WO2002020566A2/en not_active Application Discontinuation
- 2001-08-28 JP JP2002525185A patent/JP2004510710A/en not_active Ceased
- 2001-08-28 CA CA002421283A patent/CA2421283A1/en not_active Abandoned
- 2001-08-28 AU AU2002212172A patent/AU2002212172A1/en not_active Abandoned
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4923818A (en) * | 1987-09-04 | 1990-05-08 | Washington University | DNA clone of human type IV collagenase |
WO1998012309A2 (en) * | 1996-09-17 | 1998-03-26 | Washington University | Method of cell surface activation and inhibition |
Non-Patent Citations (1)
Title |
---|
P C BROOKS ET AL.: "Disruption of angiogenesis by PEX, a noncatalytic metalloproteinase fragment with integrin binding activity" CELL, Bd. 92, 6. Februar 1998 (1998-02-06), Seiten 391-400, XP002195424 CELL PRESS, CAMBRIDGE, NA., US ISSN: 0092-8674 in der Anmeldung erw{hnt * |
Also Published As
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JP2004510710A (en) | 2004-04-08 |
CA2421283A1 (en) | 2003-03-05 |
EP1315804A2 (en) | 2003-06-04 |
WO2002020566A3 (en) | 2002-06-27 |
DE10044325A1 (en) | 2002-03-21 |
AU2002212172A1 (en) | 2002-03-22 |
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