Bitenova derivatives, method of production thereof, pharmaceutical compositions and method of production thereof

 

(57) Abstract:

Describes the new bitenova compounds of General formula I where Q is absent or represents-NH-(CH2)5-CO-, R1denotes X-Arq-Gly-Asp-y, X = Tripeptide: Gly-Gly-Gly-, y = dipeptide: -Ser-Pro, or R1denotes A-Cys(R2)-B-U, R2denotes H, Trt; And represents Asp or peptide fragment selected from the group consisting of: Ala-Asp, Thr-Ala-Asp, Lys - Thr-Ala-Asp, Lys - Ala-Ala-Asp, Arq-Thr-Ala-Asp, Ser-Ala-Asp, Gln-Ser - Ala-Asp. Gly-Lys-Thr - Ala-Asp, Ile-Ser-Ala-Gly, Arq-Ser-Ala-Gly-Lys-Thr-Cys(Trt)-Asp, is absent or represents Pro or N-methylated derivative of Ala, and, if R1denotes A-Cys(R2)-B-U, only one of the residues a or b may be absent, U indicates He or NH-2or R1means cyclo(Arq-Gly-Asp-Z), Z in the side chain is linked to Q or if Q otsutstvuet, with Biotin, z denotes a di - or Tripeptide residue, and amino acids independently from each other selected from the group consisting of Ala, Val, Tyr, Trp, Phe, Cys, Lys, M, these amino acids can be derivatization, and amino acid residues are linked to each other peptidome through N-amino and-carboxyl groups, moreover, M is contained and always denotes NH(R8)-CH(R3)-COOH, R3= R6- R4where R4= HE, R6=active amino acids and amino acid derivatives, it included, as D-and L-forms and their salts. Describes the method of obtaining, pharmaceutical compositions and method of production thereof. The connection can be used as integranova inhibitors, in particular, for the prevention and treatment of diseases of the circle of coopermine, thrombosis, heart attack, arteriosclerosis and therapy of tumors. 4 C. and 1 C.p. f-crystals, 1 table.

The invention relates to biotinyl compounds of General formula

< / BR>
where Q is absent or represents-NH-(CH2)5-CO-,

R1denotes X-Arg-Gly-Asp-y,

X - Tripeptide: Gly-Gly-Gly-,

y - dipeptide: -Ser-PrO

or R1denotes A-Cys(R2)-B-U,

R2denotes H, Trt;

And represents Asp or peptide fragment selected from the group consisting of:

Ala-Asp, Thr-Ala-Asp, Lys-Thr-Ala-Asp, Lys-Ala-Ala-Asp,

Arg-Thr-Ala-Asp, Ser-Ala-Asp, Gln-Ser-Ala-Asp,

Gly-Lys-Thr-Ala-Asp, Ile-Ser-Ala-Gly, Arg-Ser-Ala-Gly

Gly-Lys-Thr-Cys (Trt)-Asp

In tstststs or represents Pro or N-methylated derivative of Ala,

moreover, if R1denotes A-Cys(R2)-B-U, only one of the residues a or b may not appear,

U indicates HE or NH2,

or R1means cyclo (Arg-Gly-Asp-Z), Z in the side chain is linked to Q, or if Q is absent, with Biotin,

Z PY, consisting of Ala, Val, Tight, Trp, Phe, Cys, Lys, M, these amino acids can be derivatization, and amino acid residues are linked to each other peptidome through N-amino and-carboxyl group, and M always contains and refers to:

NH(R8) - CH(R3) - COOH,

R3= R6- R4< / BR>
where R4= HE

R6= alkylether with 7-14 C atoms,

R8= H, alkyl with 1-6 C-atoms,

moreover, if we are talking about the balance of optically active amino acids and amino acid derivatives, are included as D-, L-forms;

and their salts.

Such compounds biotinylated peptides are described, for example, WIPO 9415956 (biotinylated antagonists endothelioma receptor), WIPO 9413313 (biotinylated LHRH - antagonists) or in WIPO 9418325 (biotinylated necrosis factor).

Biotinylation peptides during synthesis in the solid phase resin with the aim of improved purification described by T. J. Lobi and others in Anal.Biochem. 170, 502 (1988).

Such compounds of cyclic and linear peptides known from patents Germany 43 10 643 and 43 36 758, European patent EP 0 406 428 and WIPO 89/05150.

The basis of the invention is to obtain new connection is STV.

It was found that the compounds of formula I and their salts with good endurance possess very valuable pharmacological properties. First of all, they act as integrin inhibitors, and, in particular, they inhibit interactionv-,3or5- integranova receptors with ligands, such as, for example, the binding of fibrinogen3- integranova receptor. Particularly effective compounds exhibit in the case of integrinsv3,v5,in3andv1,v6andv8. This action can be detected, for example, by the method described by J. W. Smith and others in J. Biol. Chem., 265, 12267- 12271 (1990).

The dependence of the occurrence of angiogenesis from the interaction between vascular integrins and extracellular matrix proteins described by P. C. Brooks, R. A. Clark, and D. A. Cheresh in Science 264, 569-71 (1994).

The possibility of inhibition of this interaction and with it the beginning of apoptosis (programmed death of cells) angiogenic vascular cells by cyclic peptide described by P. C. Brooks, A. M. Montgomery, M. Rosenfeld, R. A. Reisfeld, T.-Hu, G. Klier and D. A. Cheresh in the Cell, 79, 1157-64 (1994).

The compounds of formula I that inhibit the interaction integranova of receptorantagonists prevent the spread of tumor cells due to metastasis. This is proved by the following observations:

The spread of tumor cells from the local tumor in the vascular system occurs due to the formation of microaggregates (microthrombi) due to the interaction of tumor cells with platelets. Tumor cells escaped through protection microaggregate and are not recognized by cells of the immune system. Microaggregate may be embedded in the walls of blood vessels, making easier the further penetration of tumor cells into the tissue. Since the formation of microthrombi promotes the binding of fibrinogen with fibrinogen receptors on activated platelets, GP IIa/IIIb antagonists can be considered as effective inhibitors of metastasis.

The compounds of formula I can be used as biologically active substances in medicinal products in medicine and veterinary medicine, in particular for the prophylaxis and/or therapy of diseases such as thrombosis, myocardial infarction, arteriosclerosis, inflammation, apoplexy, angina, neoplastic disease, osteolytic diseases, such as osteoporosis, pathological angiogenic diseases, such as inflammation, eye disease, diabetic retinopathy, is t, osteoarthritis, robotically glaucoma, ulcerative colitis, Crohn's disease, atherosclerosis, psoriasis, restenosis after angioplasty, viral infection, bacterial infection, fungal infection, acute renal failure and during wound healing to maintain the healing process.

The compounds of formula I can be used as antimicrobe active substances in operations where used biomaterials, implants, catheters or electrostimulation of the heart. They act bacteriostatically. The effectiveness of antimicrobial activity can be detected using the method described by R. Valentin-Weigund and others in Infection and Immunity, 2851-2855 (1988).

Since the compounds of formula I are inhibitors of the binding of fibrinogen and together with these ligands fibrinogen receptors with platelets, they can be used in vivo as a diagnostic tools to identify and localization of thrombi in the vascular system, as bitingly the rest is a UV - detectable residue.

The compounds of formula I can be used as inhibitors of the binding of fibrinogen, as well as effective therapeutic agents for metabolic studies of platelets in razlika unit bitenova label" allows after binding to the receptor, to explore these mechanisms.

Above - and below-reducing amino acid residues indicate residues of the following amino acids:

Abu - 4-aminobutyric acid; Aha - 6-aminohexanoic acid; 6-aminocaproic acid; Ala, alanine; Asn, asparagine; Asp - aspartic acid; Arg, arginine; Cys - cysteine; Dab - 2,4-diaminobutane acid; Dab - 2,3-diaminopropionic acid; Gln - glutamine; Glp - pyroglutamyl acid; Glu - glutamic acid; Gly is glycine; His - histidine; Homo-Phe - Homo-phenylalanine; IIe - isoleucine; Leu - leucine; Lys, lysine; Met, methionine; Nle - norleucine; Orn is ornithine; Phe is phenylalanine; Phg - phenylglycine; 4-al-he - 4-halogen-phenylalanine; Pro - Proline; Ser - serine; Thr - threonine; Trp: tryptophan; Tyr, tyrosine; Val is valine; and Bit:

< / BR>
Here are the following notation:

VOS - tert.-butoxycarbonyl; CBZ - benzyloxycarbonyl; DCCI - dicyclohexylcarbodiimide; DMF is dimethylformamide; Et is ethyl; Fmoc- 9-fluorenylmethoxycarbonyl; HOBt- 1-hydroxybenzotriazole; Me is methyl; MBNA - 4-methyl-benzhydrylamine; Mtr - 4-methoxy-2, 3,6-trimetilfenil-sulfonyl; OBut - complex tert. -butyl ether; OMe-methyl ester; OEt - complex ethyl ester; ROA - phenoxyacetyl; F (TPA) - triperoxonane enantiomeric forms, it includes all above - and below these forms and also their mixtures (for example, DL - form), for example, as an integral part of the compounds of formula I. Further, amino acids, for example, as an integral part of the compounds of formula I, can be provided with the relevant itself known protective groups.

Proposed according to the invention compounds also include the so-called proletarienne derivatives, i.e. modified, for example, with alkyl or acyl groups, sugars or oligopeptides of the compounds of formula I, in which the body quickly split up effective, proposed according to the invention compounds.

The subject invention further is a method of obtaining compounds of formula I on p. 1 of the claims, as well as their salts, characterized in that

(a) compound of formula II: H-Q-R1II,

where Q and R1are specified in paragraph 1 of the claims is by the reaction of acylation enter into interaction with the compound of the formula III:

< / BR>
where L denotes Cl, Br, J, or a free or reactive functionally modified OH group and, if necessary, the base or acidic compound of formula I by arr is and L have indicated in the case of formulas II and III values, unless nothing else.

In the above formulas, the alkyl preferably denotes methyl, ethyl, isopropyl or tert.-butyl.

Alkylene preferably represents methylene, ethylene, propylene, butylene, pentile or hexylen.

Alkylammonium is preferably benzyl or phenethyl.

Alkylenediamine preferably represents 4-methylene-benzyl or 4-ethylene-benzyl.

The remainder is R6-R4preferably represents 2-, 3 - or 4-hydroxybenzyl; 2-, 3 - or 4-hydroxyphenethyl.

These values of X, Y and Z are amino acids and amino acid residues can also be derivatization, preferably N-methyl, N-ethyl, N-sawn, N-benzyl or C-methyl derivatives.

Hereinafter, preferred derivatives of Asp and Glu, in particular complex of methyl, ethyl, propyl, butyl, tert.- butyl, neopentyl or benzyl esters of the carboxyl groups of the side chains; further, derivative Arg, which in-NH - C(=NH)-NH2group may be substituted by acetyl, benzoline, methoxycarbonyl or ethoxycarbonyl balance. Next, specified in Zvezdnyi protective groups.

Z preferably denotes M; further preferably denotes a sequence of N 1 (see the end of the description).

The compounds of formula I may have one or more chiral centers and therefore may exist in different stereoisomeric forms. Formula I encompasses all these forms.

Accordingly, the subject invention are in particular those compounds of formula I in which at least one of these residues has one of the abovementioned preferred meanings. Some preferred groups of compounds can be expressed by the following partial formulae Ia to Ie, which correspond to the formula I and where more is not specified residues have the data in the case of formula I where, however,

in Ia: Q is absent and R1denotes X-Arg - Gly-Asp-Y;

in IB: Q denotes-NH-(CH2)5-CO - and R1denotes X-Arg-Gly-Asp-Y;

in IB; Q represents-NH-(CH2)5-CO - and R1means cyclo-(Arg-Gly-Asp-Z);

in Iك: Q denotes-NH-(CH2)5-CO and R1means cyclo-(Arg-Gly-Asp-M);

in Ia: Q denotes-NH-(CH2)5-CO - and R1denotes A-Cys (R2); And

in S: Q denotes-NH-(CH2)n-CO; R1denotes X-Arg-Gly-Asp-Y and "n" on rochem itself known methods, which are described in the literature (such as Houben-Weil, Methods of organic chemistry, ed. Georg-Thieme, Stuttgart), namely under reaction conditions which are known and suitable for the specified interactions. You can also use themselves known here more not mentioned options.

The source of the substance, if desired, can also be obtained in situ, so they are not isolated from the reaction mixture, and immediately injected into the interaction further, to obtain compounds of formula I.

The compounds of formula I can preferably be obtained by the fact that the compounds of formula II is administered in cooperation with compounds of formula III.

Compounds of formulas II and III are generally known. If they are unknown, they can get itself known methods.

In compounds of formula III residue-CO-L, indicates a pre-activated carboxylic acid, preferably galoyanized carboxylic acid, a symmetrical or mixed anhydride or activated complex ester. Such residues to activate carboxyl groups in a typical acylation reactions described in the literature (for example, in Houben - Weil, Methods of organic chemistry, ed. Georg - Thieme, Stuttgart).

L preferably denotes H, Cl, Br or-ON - succinimide.

The interaction is carried out usually in an inert solvent, in the presence of cyclotosaurus means, preferably organic bases like triethylamine, dimethylaniline, pyridine or quinoline, or an excess of the carboxyl component of the formula III.

It may also be favorable additive hydroxide, carbonate or bicarbonate of alkali or alkaline earth metal or of another salt of a weak acid of the alkali or alkaline earth metal, preferably potassium, sodium, calcium or caesium.

The reaction time, depending on the applied conditions, ranges from a few minutes up to 14 days; the reaction temperature is from -30oC to + 140oC, usually from -10oC to + 90oC, in particular from about 0oC to about 70oC.

As inert solvents are suitable, for example, hydrocarbons as hexane, petroleum ether, benzene, toluene or xylene; chlorinated hydrocarbons, such as trichloroethylene, 1,2-dichloroethane, carbon tetrachloride, chloroform or dichloromethane; alcohols, like methanol, ethanol, isopropanol, n-propanol, n-butanol or tert.-butanol; ethers, as IER is colmenarejo and-monotropy simple ether(methylglycol or ethylglycol), etilenglikolevye simple ether (diglyme); ketones, such as acetone or butanone; amides, as ndimethylacetamide, dimethylacetamide or dimethylformamide (DMF); NITRILES like acetonitrile; sulfoxidov as dimethyl sulfoxide (DMSO); carbon disulfide; carboxylic acids, as formic acid or acetic acid; nitro compounds, as nitromethane or nitrobenzene; esters like ethyl acetate, water, or mixtures of these solvents.

The compounds of formula I, further, can be obtained by the fact that the compounds of formula IY enter into an interaction with compounds of formula Y. the starting compound of formula IV, and formula Y generally known. If they are unknown, they can get itself known methods.

In the compounds of formula Y-balance-CO-L denotes pre-activated carboxylic acid, preferably galoyanized carboxylic acid, a symmetrical or mixed anhydride or activated ester. Such residues to activate carboxyl groups in a typical acylation reactions described in the literature (for example, Houben-Weil, Methods of organic chemistry, ed. Georg - Thieme, Stuttgart).

L preferably denotes Cl, Br or-ON-succinimide.

Interaction with the no, and temperature, and solvent, as described for the interaction of the compounds of the formula II with compounds of formula III.

The linear compounds of formula I with an open circuit, in which R1denotes X-Arg-Gly-Asp-Y or A-Cys(R2)-B, then you can get the fact that in the last stage of solid-phase synthesis of Biotin is introduced into the reaction mix in the same cycle as conventional protected at N-end amino acid, as the last component and biotinylated under normal conditions otscheplaut from resin. Solid-phase synthesis, cleavage and purification carried out as described by A. Jonczyk and J. Meienhofer in Peptides. Proc. 8th Am. Pept. Symp. Eds. V. Hruby and D. H. Rich, Pierce Comp III, S. 73-77 (1983), or analogously to the methods described in Angew. Chem. 104, 375-391 (1992).

A linear connection with an open chain of formulas II and IV, however, you can get the usual methods of amino acid and peptide synthesis, which, for example, described in these standard works and patent applications, for example, by solid phase synthesis according to Merrifield (B. F. Gysin and R. B. Merrifield, J. Am.Chem.Soc., 94, 3102 and later (1972)).

The cyclic compounds of formula II and formula IV, in which R1means cyclo-(Arg-Gly-Asp-Z), can be obtained by cyclization of linear compounds, as, for example, described is about to get by their release from their functional derivatives by solvolysis, in particular hydrolysis, or by hydrogenolysis.

The preferred initial agents for the solvolysis, respectively, hydrogenolysis are those which, instead of one or more free amino and/or hydroxyl groups contain corresponding protected amino and/or hydroxyl groups, preferably such that instead of H-atom with a N-atom, are protective for the amine function group, for example, those that correspond to the formula I, but instead of NH2groups contain other1group (where R1means for protective amine function group, such as BOC or CBZ).

Hereinafter, the preferred source of substances that instead of the H atom of the hydroxyl group containing protective for hydroxyl function group, for example, those that correspond to the formula I, but instead hydroxyproline groups contain R"O-phenyl group (where R' denotes a hydroxyl protective for function group).

In the molecule of the original substance may be several identical or different protected amino and/or hydroxyl groups.

If the existing protective g is Etna for the amine function group" is well known and refers to groups which are suitable for protecting (blocking) an amino group from chemical interactions, which, however, easily hatshepsuts after in other parts of the molecule was desired chemical reaction. Typical of such groups are, in particular, unsubstituted or substituted acyl, aryl, arelaxation or kalkilya group. As for protective amine function group is removed after the desired reaction (or sequence of reactions), their race and size is not critical; however, a preferred group with 1-20, in particular 1-8, C atoms. The expression "acyl group" in connection with the present method should be understood in its broadest sense. It covers produced from aliphatic, alifaticheskih, aromatic or heterocyclic carboxylic acids or sulfonic acids acyl group, and, in particular, alkoxycarbonyl, aryloxyalkyl and primarily alcoxycarbenium group. Examples of such acyl groups are alkanoyl as acetyl, propionyl, butyryl; arcanol as phenylacetyl; aroyl as benzoyl or toluyl; aryloxyalkanoic as POA; alkoxycarbonyl as methoxycarbonyl, etoxycarbonyl and 2.2.2-trichlorocyanuric, VOS, 2-iodoxybenzoic; Aral is elegance, protective for amine-function groups are BOC and Mtr next, CBZ, Fmoc, benzyl and acetyl.

The expression "hydroxyl protective for function group" is also generally known and relates to groups which are suitable for protecting a hydroxyl group from chemical interactions, which, however, can be easily removed, after in other parts of the molecule was desired chemical reaction. Typical of such groups are the abovementioned unsubstituted or substituted aryl, kalkilya or acyl group; hereinafter, also alkyl groups. The nature and magnitude protective for hydroxyl group functionality is not critical, as they again removed after the desired chemical reaction or sequence of reactions; preferred group with 1-20, in particular 1-10, C atoms. Examples of hydroxyl protective for function groups are, inter alia, benzyl, p - nitrobenzoyl, p-toluensulfonyl, tert.-butyl and acetyl, and especially preferred benzyl and tert.-butyl. COOH-group in aspartic acid and glutamic acid are preferably protected in the form of their complex tert.-butyl esters (for example, ASR (but)).

Release of the compounds of the formula I from their functional derivatives by gostoso TFA or perchloric acid, however, also using other strong inorganic acids as hydrochloric acid or sulfuric acid, strong organic carboxylic acids, as trichloroacetic acid, or sulfonic acids, as benzene - or p-toluensulfonate. The presence of an additional inert solvent may, but is not always necessary. As inert solvents are suitable, preferably organic, for example carboxylic acids, as acetic acid; ethers, like tetrahydrofuran or dioxane; amides as DMF; halogenated hydrocarbons like dichloromethane; hereinafter, also alcohols as methanol, ethanol or isopropanol, and also water. Next, apply a mixture of the above solvents. TFA is preferably used in excess without the addition of another solvent; perchloric acid is used in the form of a mixture of acetic acid and 70% perchloric acid in the ratio 9:1. Reaction temperatures for the cleavage of the appropriate amount from about 0oup to about 50oC, preferably operate at a temperature of 15-30oC (room temperature).

Group BOC, OBut and Mtr can be split, for example, preferably using TFA in dichloromethane or using approximately 3 to 5 N. HCl in dioxane at 15-30oC; F in DMF at 15-30oC.

Trityloxy group used to protect the amino acids histidine, asparagine, glutamine and cysteine. Removal is carried out, depending on the desired target product, using TFA/10% thiophenol, and triticina group is cleaved from all of these amino acids, when using a mixture of TFA/anisole or TFA/thioanisole cleaved only triticina group from His, Asn, and Glu, as opposed so that it remains in the Cys side chain.

Hydrogenations removable protective group (for example, CBZ or benzyl) can be split, for example, by treatment with hydrogen in the presence of a catalyst (for example, a catalyst based on a noble metal, such as palladium, expediently on the media, as coal). As solvents if this is suitable to the above, in particular, for example, alcohols, like methanol or ethanol, or amides, as DMF. Hydrogenolysis, as a rule, is carried out at a temperature of about 0-100oC and pressures of about 1-200 bar, preferably at 20-30oC and a pressure of 1-10 bar. Hydrogenolysis of CBZ - group flows, for example, a well in the presence of 5-10% palladium-on-coal in methanol or using ammonium formate (instead of hydrogen) in the presence of Pd/C in a mixture of methanol with DMF PR is unity acid, for example, by reacting equivalent amounts of base and acid in an inert solvent like ethanol and subsequent evaporation. For this interaction is used, in particular, acids, which give physiologically acceptable salts. Thus, it is possible to use inorganic acids, for example sulfuric acid, nitric acid, halogen acids as hydrochloric acid or Hydrobromic acid, phosphoric acid, like phosphoric acid; sulfamic acid; further, organic acids, in particular aliphatic, alicyclic, analiticheskie, aromatic or heterocyclic one - or polybasic carboxylic, sulfonic or sulfuric acids, such as formic acid, acetic acid, propionic acid, pavlikova acid, diethyloxalate acid, malonic acid, succinic acid, Emelyanova acid, fumaric acid, maleic acid, lactic acid, tartaric acid, malic acid, citric acid, gluconic acid, ascorbic acid, nicotinic acid, isonicotinamide acid, methane - or econsultation, ethicalfashion, 2 - hydroxyethanesulfonic, benzosulfimide, p - toluensulfonate, naphthalene mono - and di-criminate for isolating and/or purifying compounds of formula I.

On the other hand, the acid of formula I by entering into interaction with the base can be translated into one of its physiologically acceptable metal salts or ammonium. As the salts used in this case, in particular, salts of sodium, potassium, magnesium, calcium and ammonium; next, substituted ammonium salts, for example, dimethyl-, diethyl - or aminobutiramida - ammonium salt; monoethanol-, diethanol - or aminobutiramida - ammonium salt; cyclohexyl, DICYCLOHEXYL-ammonium salt; dibenzylethylenediamine salt; further, for example, salts with arginine or lysine.

The subject of the invention, then, is the use of compounds of the formula I and/or their physiologically acceptable salts for the preparation of pharmaceutical compositions, in particular, non-chemical way. Them together, at least one solid, liquid and/or semi-liquid carrier or auxiliary substance and, if necessary, in combination with one or more other biologically active substances brought to a suitable dosage forms.

The subject of the invention, further, are pharmaceutical compositions containing at least one compound of the formula I and/or one of its physiologically acceptable salts.

2or perchloroethane). It is advisable to apply the biologically active agent in micronized form, and may be added one or more additional physiological acceptable solvents such as ethanol. Solutions for inhalation can be entered using a conventional inhalers.

The compounds of formula I and their physiologically acceptable salts can be used as integrin inhibitors for combating diseases, in particular pathological angiogenic diseases, thrombosis, heart attack, coronary heart disease, arteriosclerosis, tumors, osteoporosis, inflammations and infections.

This proposed according to the invention substances, as a rule, you can enter by analogy with other known, commercially available peptides, in particular, however, similar to the one described in the U.S. patent And 4472305 compounds, preferably in dosages of about 0.05 to 500 mg, in particular 0.5 to 100 mg, dosing unit. The daily dosage is about measures the effectiveness of used special compound, the age, body weight, General health, sex, on cost, time and route of administration, rate of excretion, combination of drugs and the severity of the relevant disease, which has implications for therapy. Preferably parenteral administration.

Furthermore, the new compounds of formula I can be used in analytical biology and molecular biology. Using the ability to complexation between biotinyl balance and glycoprotein by Avidya. Application of a complex of Biotin-avidin is known from the article by E. A. Bayer and M. Wilchek in Methods of Biochemical Analysis, 26, 1-45 (1980) [1].

The new compounds of formula I can be used as integranova ligands for the preparation of columns for affinity chromatography to obtain pure integrin. The complex of derivatizing Avidya media, such as sepharose, and new compounds of the formula I obtained in itself known methods as described, for example, in [1]. For this reason, there are not further dwell on this method, refer to the relevant literature, for example [1].

As polymeric carriers suitable the actual CE is sewn, polisher, as cellulose, sepharose or Sephadexacrylamide, polymers based on polyethylene glycol or Tentakel-polymers< / BR>
The new compounds of formula I can also be used as diagnostic markers for anti-Biotin-antibody reactions in the analysis of enzyme immunosorbent type and FACS analysis (fluorescence activated installation for sorting cells). Known application of antibiotin antibodies for the detection of Biotin according to M. Berger, Biochemistry, 14, 2338-2342 (1975). Application derivatizing Biotin immunoglobulin lqG in the enzyme-linked immunosorbent assay (ELISA) described in U. Holmkov-Nielsen and others, in Journal of Chromatogr., 297, 225- 233 (1984).

In J. Immunol.Methods., 181, 55-64 (1995) J. Gao, and S. J. Shattil describes ELlSA test for the detection of substances which inhibit the activation of the integrin. For detection used here biotinylated fibrinogen.

Application of flow cytometry in clinical diagnosis of the cells described G. Schmitz, and G. Rothe in DG Klinische Chemie], 24 (1993) part 1, S. 1 - 14.

Further, the compounds of formula I can be used in microscopy with a force field (atomic force microscopy F) to measure the strength of interactions of the ligand-receptor. The ligand preferably denotes a complex of avidin and other in Science 264, 415 - 417 (1994) described the measurement of the adhesion forces between the functionalized by Avidya microscope with a force field and biotinylated agarose.

Above and below, all temperatures are indicated inoC. In the following examples, "conventional treatment" means add, if necessary, water; establish, if necessary, depending on the structure of the target product, the pH values in the range 2-10; extracted with ethyl acetate or dichloromethane, separated, the organic phase is dried over sodium sulfate, the solvent is evaporated and the residue purified by chromatography on silica gel and/or by crystallization. Rf - values are specified using silica gel; solvent: a mixture of ethyl acetate with methanol in the ratio 9:1.

RZ - retention time (minutes) in the case of HPLC (high performance liquid chromatography) under the following systems:

[A]:

Column: Nucleosil 7C 18 h mm; eluent A: 0.1% of TFA in water; eluent B: 0.1% of TFA in acetonitrile; after: 1 ml/min; gradient: 20-50% B /30 min

[B]:

50-min gradient of 0-80% propan-2-ol in water with 0.3% TFA at 1 ml/min, column LichrosorbRP Select B (7 μm) h mm

[C]:

Column: Lichrospher (5 μm) 100 RP 8 h mm; eluent A: 0.0 V/50 minutes

Mass spectrometry (MS): EI (ionization by electron impact)+FAB (fast atom bombardment) (M+N)+< / BR>
DMPP - resin refers to 4-(2',4'-acid-hydroxymethyl) paroxysmal; smirkily labile (unstable) resin, which allows the synthesis of protected side chains of peptides.

Examples see the end of the description.

1. Bitenova compounds of General formula I

< / BR>
where Q is absent or represents-NH-(CH2)5-CO-;

R1denotes X-Arq-Gly-Asp-y;

X - Tripeptide: Gly-Gly-Gly-,

y - dipeptide: -Ser-PrO,

or R1denotes A-Cys(R2)-B-U,

R2denotes H, Trt;

A represents Asp or peptide fragment selected from the group consisting of: Ala-Asp, Thr-Ala-Asp, Lys-Thr-Ala-Asp, Lys-Ala-Ala-Asp, Arq-Thr-Ala-Asp, Ser-Ala-Asp, Gln-Ser-Ala-Asp, Gly-Lys-Thr-Ala-Asp, Ile-Ser-Ala-Gly, Arq-Ser-Ala-Gly, Gly-Lys-Thr-Cys(Trt)-Asp,

B is absent or represents Pro or N-methylated derivative of Ala, and, if R1denotes A-Cys(R2)-B-U, only one of the residues A or B may be absent;

U denotes OH or NH-2;

or R1means cyclo(Arq-Gly-Asp-Z), Z in the side chain of communication with Q or if Q is absent, with Biotin;

z denotes a di - or tripeptides the rest,

and amino acids, the s can be derivatization, and amino acid residues are linked to each other peptidome through N-amino and-carboxyl group, and M always contains and refers to:

NH(R8)-CH(R3)-COOH,

R3= R6- R4,

where R4= OH,

R6= alkylether from 7 to 14 C-atoms;

R2= H, alkyl with 1 to 6 C-atoms,

moreover, if we are talking about the balance of optically active amino acids and amino acid derivatives, are included as D-and L-forms and their salts.

2. Bitenova the compounds of formula under item 1

< / BR>
where (a) Q is absent and R1means cyclo-(Arq-Gly-Asp-D-Phe-Lys); b) Q is absent and R1denotes Gly-Gly-Gly-Arq-Gly-Asp-Ser-Pro-Lys-OH; Q is absent and R1denotes Gly-Gly-Gly-Lys-Thr-Ala-Asp-Cys(Trt)-Pro-OH; g) Q denotes-NH-(CH2)5-CO - and R1means cyclo-(Arq-Gly-Asp-D-Phe-Lys); d) the Q denotes-NH-(CH2)5-CO - and R1means cyclo-(Arq-Gly-Asp-D-Phe-Lys-Gly); (e) the Q denotes-NH-(CH2)5-CO - and R1means cyclo-(Arq-Gly-Asp-D-Phe-Val-Lys); W) Q denotes-NH-(CH2)5-CO - and R1means cyclo-(Arq-Gly-Asp-D-Trp-Lys); C) Q denotes-NH-(CH2)5-CO - and R1means cyclo-(Arq-Gly-Asp-D-Tyr-Lys),

as well as physiologically acceptable salts of these compounds.

3. The way polur>,

where Q and R1have the values listed in paragraph 1,

acelerou the compounds of formula III

< / BR>
where L denotes CL, Br, J, or free, or reactive functionally modified OH-group,

and, if necessary, the base or acidic compound of formula I by treatment with an acid or a base is translated into one of its salts.

4. The method of obtaining pharmaceutical compositions having the properties of receptor antagonists of adhesion, characterized in that an effective amount of the compounds of formula I under item 1 and/or one of its physiologically acceptable salts together with at least one solid, liquid or semi-liquid carrier or auxiliary substance is brought to a suitable dosage forms.

5. Pharmaceutical composition having the properties of receptor antagonists of adhesion, characterized in that it contains at least one compound of formula I under item 1 and/or one of its physiologically acceptable salt in an effective amount.

 

Same patents:

-(3-n-methylpyridin s)-alanine as-(l)- rarn-Сhg-Рal iu(3)-nh2" target="_blank">

The invention relates to a new method of obtaining N-acetyl-(L)-4-cyanopyrrolidine separation of the racemate ethyl ester of N-acetyl-(D,L)-4-cyanopyrrolidine and to a new method of obtaining stereoisomer Ac-(L)-pAph-Chg-Me Pal(3)-NH2using as an intermediate compound N-acetyl-(L)-4-cyanopyrrolidine

The invention relates to products derived from histamine and, in particular, the condensation products of histamine or methylsiloxanes histamine and amino acids, the method of their preparation and use as active principle in areas such as therapy and cosmetology, as well as the factor (agent), improving the stability of compositions used in therapy, cosmetology, agriculture and food industry (region)

The invention relates to biotechnology and can be used for Introduzione nucleic acids into cells

The invention relates to bicyclic compounds useful as drugs, the neutralizing effect of glycoprotein IIb/IIIa, to prevent thrombosis

The invention relates to medicine and can be used to adjust parameters of the system of hemostasis

The invention relates to a drug intended for intravenous

The invention relates to new cyclic amine derivatives of General formula I, where R1represents a phenyl group substituted by halogen atom,2represents C1- C8aliphatic acyl group or (C1- C4alkoxy)carbonyl group, R3represents a 3 - to 7-membered saturated cyclic amino group which may form a condensed ring, where the specified cyclic amino group substituted by the Deputy selected from the group comprising: mercaptopropyl, which can be unprotected or protected by a group selected from a number of protective groups, C1- C4alkyl group, substituted mercaptopropyl, which can be unprotected or protected by a group selected from a number of protective groups, and the number of protective groups for the specified mercaptopropyl includes C1- C20alcoholnye group, C3- C20alkenone group and benzoline group, and the said cyclic amino group, furthermore preferably a substituted group of the formula =CR4R5where R4represents a hydrogen atom, and R5represents a hydrogen atom, a C1- C4alkyl group, carboxypropyl, (C1- C4-alkoxy)carbonyl GRU

The invention relates to medicine

The invention relates to a new derived in a series of 1H-imidazo[1,2-a]benzimidazole, namely water-soluble dihydrochloride of 1-(2-isopropylaminoethyl)-2-phenylimidazo[1,2-a]benzimidazole of the formula I

< / BR>
with a local anesthetic action at a terminal, infiltration and block anesthesia
Up!