Derivative peptide, or their salts, the pharmaceutical composition

 

(57) Abstract:

Usage: in medicine, as new peptides provide potentially improved therapeutic treatment of neoplastic diseases. The essence of the invention: derivatives of the peptide of General formula I: R1R2N-CH(X) CO-A-B-D-(E)u-(F)v(G)w-K, where R1- AlkO, (ness.) Alk, FAlk, cyclo Alk, aminosulfonyl, the latter may be substituted Alk, OH, or NR3R4where each R3and R4can be hydrogen or alkyl; R2is hydrogen, Alk, FAlk, cyclo Alk, R1and R25-membered heterocycle, a - amino acid residue selected from Val, Ile, Leu, ALLO-Ile, -aminoisobutyrate, 3-tert-butyl Ala, 2-tert-butyl Gly, 3-cyclohexyl Ala, 2-ethyl Gly, 2-cyclohexyl Gly, Nle, NVal; B is the remainder: N-Alk-Val, NVal, Leu, Ile, 2-tert-butyl Gly, 3-tert-butyl Ala, 3-cyclohexyl Ala, Phe, 2-cyclohexyl Gly; D, E, F and G, independently, is chosen from the group consisting of: Pro, Homo-Pro, hydroxy-Pro, thiazolidine-4-carbonyl, 1-aminopentyl-1-carbonyl, Val, 2-tert-butyl Gly, Ile, Leu, 2-cyclohexyl Ala, Phe, N-methyl Phe, tetrahydroisoquinoline-2-carbonyl, 3-thiazolyl Ala, 3-thienyl Ala, His, 1-aminoindane-1-carbonyl, Arg, 3-pyridyl Ala, 3-tert-butyl Ala, 2-cyclohexyl Gly, NVal, NLeu and 3-naphthyl Ala; X is hydrogen, alkyl, cycloalkyl-CH2the, sulfur; u, v and w are equal, independently, 0 or 1; K - OH, AlkO, phenoxy, benzyloxy, substituted or unsubstituted amino group, or their salts with physiologically compatible acids; a pharmaceutical composition comprising a carrier and an active ingredient of the formula I in an effective amount. 2 C. and 8 C.p. f-crystals, 8 tab., 74 ill.

The invention relates to medicine.

The invention offers new derivatives of peptides exhibiting antitumor activity, with potentially improved therapeutic effect in the treatment of neoplastic diseases compared with dolastatin-10 and -15 (U.S. patent 4879278, November 7, 1989; U.S. patent 4816444, March 28, 1989). In addition, unlike dolastatin-10 and -15, which must be allocated by the laboratory of rare natural sources, the compounds of the present invention can be synthesized, as described in detail below. In addition, dolastatin-10 unstable to acids. Described that even small changes in structure can cause complete loss of activity (Biochemical Pharmacology, volume 40, N 8, 1859-1864, 1990).

The invention also features a pharmaceutical composition using the new derived peptides.

Compounds according to the invention vkluchil, which may be substituted by alkyl; hydroxy or NR3R4where each R3and R4can be either hydrogen or alkyl;

R2is hydrogen, alkyl, foralkyl, cycloalkyl;

R1-N-R2together may denote a 5-membered heterocycle which can be unsubstituted or substituted by one or more substituents, represents an alkyl group;

A is the residue was felled, isoleucyl, laitila, allasalueella, aminoisobutyrate, 3-tert-butylamine, 2-tert-butylphenyl, 3-cyclohexylethyl, 2-ethylglycine, 2 - cyclohexylglycine, norleucine, Norvaline;

B - N-alkylaryl, -Norvaline-Lalla-isoleucyl, -2-tert-butylglycol, -3-tert-butylamine, -3-cyclohexylurea, i.e. phenylalanyl or-2-cyclohexylglycine;

D, E, F and G independently are selected from the group consisting of prolyl residues, homepolish, hydroxyproline, thiazolidine-4-carbonyl, 1-aminopentyl-1-carbonyl, thronged, 2-tert-butylglycol, isoleucyl, laitila, 3-cyclohexylamine, i.e. phenylalanyl, N-methylphenylamine, tetrahydroisoquinoline-2-carbonyl, 3-thiazolidinone, 3-thienylene, histidine, 1-aminoindan-1-carbonyl, arginine, 3-pyridylethyl, 3-tert-butylamine, 2-cyclohexylglycine, Norvaline, together may designate

< / BR>
where Y is hydrogen or lower alkyl; Z is hydrogen or lower alkyl; n = 1, 2, or 3; V is oxygen or sulfur; M is hydrogen, lower alkyl, aralkyl, cyclohexyl, or-CH2-cyclohexyl; Q is hydrogen; R is hydrogen or lower alkyl; or R and Q can together form a bond; U is hydrogen, lower alkyl, phenyl or cycloalkyl; W is hydrogen, lower alkyl or phenyl;

u, v and w are independently 0 or 1; and

K - hydroxy, alkoxy, phenoxy, benzyloxy or substituted or unsubstituted amino group;

or their salts with physiologically compatible acids.

The most preferred peptides of formula I, where R1-N-R2is a 5 - membered heterocycle of the formula

< / BR>
which may be unsubstituted or substituted by one or more substituents, represents an alkyl group.

Particularly preferred peptides of formula I, where K is the amino group of the formula R5-N-R6in which R5is hydrogen, lower alkyl, lower alkoxy, benzyl; R6is hydrogen, lower alkyl, phenyl, C5-cycloalkyl, benzyl, unsubstituted or substituted three times lower alkoxygroup, biphenyl, thiadiazolyl, substituted lower alkyl, or-CH2R7-five-membered heteroaryl, where R7is hydrogen, benzyl, and heteroaryl in which v and w are equal to 0, and K is not hydroxy, benzyloxy, phenoxy or alkoxygroup; or peptides of formula I, where u and v are equal to 0 and K does not denote a hydroxy - or alkoxygroup; or peptides of formula I, where u, v and w are equal to 1 and K - hydroxy-, alkoxy-, phenoxy or benzyloxy, or peptides of formula I, where u and v are equal to 1, w is 0 and K - hydroxy-, alkoxy-, phenoxy or benzyloxy; or peptides of formula I, where u is l, v and w are equal to 0 and K - hydroxy-, alkoxy-, phenoxy or benzyloxy.

All proposed derivative of the peptide of General formula I possess tumor-inhibitory activity.

According to the proposed invention is also a pharmaceutical composition having a tumor-inhibiting activity, comprising the active substance and the medium in which the active substances it contains peptides of formula I in an effective amount.

One subclass of compounds of the invention includes the compounds of formula I in which R1-N-R2is a 5 - membered heterocycle of the formula

< / BR>
which may be unsubstituted or substituted by one or more substituents, independently selected from alkyl (preferably C1-C4).

Another subclass of compounds of the invention includes the compounds of formula I, where K, what do benzyloxy, or C1-C7-alkyl which may be substituted by one or more fluorine atoms, or C3-C7-cycloalkyl, or benzyl, which can have up to three substituents independently selected from CF3, nitro, C1-C7-alkylsulfonyl, C1-C4-alkoxy, phenoxy, benzoxa, halogen or C1-C4-alkyl; R6is hydrogen, C1-C7-alkyl which may be substituted by one or more fluorine atoms, or C3-C7-cycloalkyl, or phenyl (which may bear up to three substituents independently selected from CF3, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl which may form a cyclic system, C1-C4-alkoxy, phenoxy, benzox or C1-C7-alkylsulphonyl), or benzyl (which may have up to three substituents independently selected from CF3, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl which may form a cyclic system, C1-C4-alkoxy, phenoxy, benzox or C1-C7-alkylsulphonyl), or naphthyl (which may have up to two substituents, independently selected from CF3, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl, C1-C43, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl, C1-C4-alkoxy, phenoxy, benzox or C1-C7-alkylsulphonyl), or biphenyl, which may have up to two substituents, independently selected from CF3, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl, C1-C4-alkoxy, phenoxy, benzox or C1-C7-alkylsulphonyl), or triphenylmethyl (which can have up to three substituents independently selected from CF3, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl, C1-C4-alkoxy, phenoxy, benzox or C1-C7-alkylsulphonyl), or benzhydrylamine (which can have up to two substituents, independently selected from CF3, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl, C1-C4-alkoxy, benzoxa, phenoxy or C1-C7-alkylsulphonyl), or benzhydrylamine (which can have up to two substituents, independently selected from CF3, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl, C1-C4-alkoxy, benzoxa, phenoxy or C1-C7-alkylsulphonyl), or naphthylmethyl (which can have up to two substituents, independently you is socci, phenoxy or C1-C7-alkylsulphonyl), or acenaphthyl (which can have up to two substituents, independently selected from CF3, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl, C1-C4-alkoxy, benzoxa, phenoxy or C1-C7-alkylsulphonyl), or acenaphthylene (which can have up to two substituents, independently selected from CF3, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl, C1-C4-alkoxy, benzoxa, phenoxy or C1-C7-alkylsulfonyl) or pyridyl (which may have up to two substituents, independently selected from CF3, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl, C1-C4-alkoxy, benzoxa, phenoxy or C1-C7-alkylsulphonyl), or picolyl (which can have up to two substituents, independently selected from CF3, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl, C1-C4-alkoxy, benzoxa, phenoxy or C1-C7-alkylsulphonyl), or benzothiazolyl (which can have up to two substituents, independently selected from CF3, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl, C1-C4-alkoxy, benzoxa, phenoxy or C1-Cof CF3, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl, C1-C4-alkoxy, benzoxa, phenoxy or C1-C7-alkylsulphonyl), or benzimidazolyl (which can have up to two substituents, independently selected from CF3, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl, C1-C4-alkoxy, benzoxa, phenoxy or C1-C7-alkylsulphonyl), or benzoxazolyl (which can have up to two substituents, independently selected from CF3, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl, C1-C4-alkoxy, benzoxa, phenoxy or C1-C7-alkylsulphonyl), or fluorenyl (which can have up to two substituents, independently selected from CF3, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl, C1-C4-alkoxy, benzoxa, phenoxy or C1-C7-alkylsulphonyl), or aminofluorene (which can have up to two substituents, independently selected from CF3, nitro, halogen, CONHBzl, CON(Bzl)2C1-C4-alkyl, C1-C4-alkoxy, benzoxa, phenoxy or C1-C7-alkylsulphonyl), or pyrimidyl (which can have up to two substituents, independently selected from CF3, nitro, SUB>-C4-alkoxy, benzoxa, phenoxy or C1-C7-alkylsulphonyl), or 5-membered heteroaryl [which can have up to three substituents independently selected from CF3, nitro, halogen, cyano, COOMe, COOEt, thiomethyl, thioethyl, thiophenyl, picolyl, acetyl, -CH2COOEt, CONH2, CONHBzl, CON(Bzl)2C1-C4-alkyl, C3-C6-cycloalkyl, C3-C4-alkylene, forming a bicyclic system with a heterocycle, C1-C4-alkoxy, benzoxa, phenoxy, phenyl (which may have up to four substituents independently selected from nitro, CF3, halogen or C1-C4-alkyl), benzyl (which may have up to four substituents independently selected from nitro, CF3, halogen, C1-C4-alkyl, naphthyl, C1-C7-alkylsulfonyl, phenylsulfonyl or C1-C4-dialkylamino)], or-CHR7is a 5-membered heteroaryl (which can have up to two substituents, independently selected from CF3, nitro, halogen, CONHBzl, CON(Bzl)2, COOMe, COOEt, COOCH(CH3)2, CONH2, COOBzl, C1-C4-alkyl, C1-C4-alkoxy, benzoxa, phenoxy, phenyl, benzyl, naphthyl or C1-C7-alkylsulfonyl [R7is hydrogen, linear or rasvet the CH2)4-]).

This subclass includes compounds of formula I, where u, v and w are independently 0 or 1; R1, R2and X is lower alkyl, A and F - amino lower-alkyl, B - N-lower alkilirovanny amino lower alkyl, D, E, G and K have the above values. With this in mind, one group of such compounds can therefore be represented by formula II

R1R2N-CXH-CO-A-D-Pro-Pro-(F)v(G)w-K

and another formula III

R1R2N-CXH-CO-A-D-Pro-Pro-(F)v(G)w-K.

B another subclass of compounds of the invention R5-N-R6together may form a structure selected from the group consisting of

< / BR>
which can be unsubstituted or substituted by one or more substituents independently selected from the group consisting of CF3, nitro, halogen, oxo, cyano, N, N-dimethylamino, CONHBzl, CON(Bzl)2C1-C6-alkyl, C3-C6-cycloalkyl, C3-C4-alkylene, forming a bicyclic system with a heterocycle, C1-C4-alkoxy, phenoxy, benzoxa, naphthyl, pyrimidyl, COOEt, COOBzl, C3-C6-cycloalkyl, pyrrolidinyl, piperidinyl, teinila, pyrrolyl, -CH2-CO-NCH(CH3)2, -CH23, thiomethyl or the corresponding sulfoxide or sulfone, thioethyl or the corresponding sulfoxide or sulfone, C1-C4-alkyl and C1-C4-alkoxy and phenyl (which may bear up to three substituents independently selected from nitro, halogen, CF3, thiomethyl, thioethyl, C1-C4-alkyl and C1-C4-alkoxy).

Preferred compounds of formula I in which the substituents have the following meanings:

R1- ethyl, methyl, 2-foretel, 2,2-dottorati, 2,2,2-triptorelin, 2-corisoprodol, tripersonal, isopropyl, propyl, butyl, pentyl, cyclopropyl, cyclopentyl, morpholinomethyl, tert-butylamino-sulfonyl, methylaminomethyl, NH2N(CH3)2N(CH2CH3)2N[CH(CH3)2]2;

R2is hydrogen, methyl, ethyl, 2-foretel, 2,2-dottorati, 2,2,2-triptorelin, 2-corisoprodol, tripersonal, isopropyl, propyl, butyl, cyclopropyl;

or R1-N-R2together represent one of the residues shown in Fig. 1;

A, B, D, E, F, G and X have the above meanings;

u, v and w are independently 0 or 1;

A and B together represent the elements in Fig. 2-6;

F and G together represent the elements in Fig. 7-11;

R5is hydrogen, methyl, ethyl, 2-foretel, 2,2-dottorati, tripersonal, propyl, isopropyl or elements in Fig. 18;

R6is hydrogen, methyl, ethyl, 2-foretel, 2,2-dottorati, triptorelin, tripersonal, propyl, isopropyl, tert-butyl or elements in Fig. 19-27;

R5-N-CHR7is a 5-membered heteroaryl refers to elements of Fig. 28;

R5-N-R6together represent the elements in Fig. 29-33;

K - hydroxy, alkoxy (preferably C1-C4), phenoxy or benzox.

More preferred compounds in which the substituents have the following meanings:

R1- ethyl, methyl, 2-foretel, 2,2-dottorati, 2,2,2-triptorelin, 2-corisoprodol, tripersonal, isopropyl, propyl, cyclopropyl, benzyloxycarbonyl, methoxycarbonyl, lactel, methylaminomethyl, tosyl, Uriel, mesyl, N(CH3)2, amidino, methoxy, benzyl, 4-phenoxybenzyl, 4-benzyloxybenzyl or 3,4,5-trimethoxybenzyl;

R2is hydrogen, methyl, ethyl, 2-foretel, 2,2-dottorati, 2,2,2-triptorelin, isopropyl, propyl, butyl, cyclopropyl, formyl, acetyl, propionyl, pivaloyl, benzoyl or benzyl;

R1-N-R2together denote

< / BR>
A, B, D, E, F, G and X have the above meanings;

E is hydroxy, C1-C4-alkoxy or benzyloxy;

R5is hydrogen, methyl, ethyl, 2-foretel, 2,2-dottorati, propyl, isopropyl, cyclopropyl, cyclopentyl, cyclohexyl, benzyl, 4-phenoxybenzyl, 4-benzyloxybenzyl or 3,4,5-trimethoxybenzyl;

R6is hydrogen, methyl, ethyl, 2-foretel, 2,2-dottorati, propyl, isopropyl, tert-butyl, cyclopropyl, cyclopentyl, cyclohexyl, benzyl, 4-phenoxybenzyl, 4-benzyloxybenzyl, 3,4,5-trimethoxybenzyl, phenyl, 4-phenoxyphenyl, 4-benzyloxyphenyl, 3,4,5-trimethoxyphenyl or elements in Fig. 36-38;

R5-N-CHR7is a 5-membered heteroaryl means

< / BR>
R5-N-R6together represent the elements in Fig. 39-41.

These examples illustrate but do not limit the scope of the present invention.

The peptides of formula I are preferably of L-amino acids, but they may contain one or more D-amino acids.

New connections may be present in the form of salts with physiologically acceptable acids such as hydrochloric acid, citric acid, tartaric acid, lactic acid, phosphoric acid, methanesulfonate, acetic acid, formic acid, maleic acid, fumaric acid, malic acid, succinic sour is a, the mucus acid, benzoic acid, glucuronic acid, oxalic acid, ascorbic acid and acetylglycine.

New connections can be obtained by known methods of peptide chemistry. Thus, peptides can be synthesized by the sequential joining of amino acids or by condensation of the corresponding peptide fragments. When performing sequential condensation, since the C-Terminus, the peptide chain is lengthened gradually with the help of one amino acid at a time. When connecting fragments can bind fragments of different lengths, and the fragments in turn can be obtained by sequential condensation of amino acids or from themselves by joining fragments.

As in the sequential build-up circuit, and by condensation of fragments, you must associate the links through the formation of amide linkages. For this purpose, suitable enzymatic and chemical methods.

Chemical methods of obtaining the amide linkages are described in detail in the works Mller, Methods der organischen Chemie, I. XV/2, c. 1-364, Thieme Verlag, Stuttgart 1974; Stewart, Young, SOlid Phase Peptide Synthesis, c. 31-34, 71-82, Pierce Chemical Company, Rockford, 1984; Bodanszky, Klausner, Ondetti, Peptide Synthesis, c. 85-128, John Wiley & Sons, New York, 1976, and other standard works of the Idov, active esters obtained in situ or pre, the use of urethane-protected N-carboxyanhydrides amino acids and the formation of amide linkages with the use of binding agents (activators), especially dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), 1-etoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), the hydrochloride of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), n-papapostolou anhydride (PPA), N,N-bis(2-oxo-3-oxazolidinyl)-iminophosphoranes (BOP-Cl), bromo-Tris-pyrrolidinone hexa-perforate (PyBrop), diphenylphosphinite (DPPA), reagent Castro (BOP, PyBop), O-benzotriazolyl-N,N,N',N'-tetramethyluronium salts (HBTU), diethylphosphoramidite (DEPCN), 2,5-diphenyl-2,3-dihydro-3-oxo-4-hydrometeorogical (reagent Steglich; HOTDO) and 1,1'-carbonyldiimidazole (CDI). Linking reagents can be used alone or in combination with additives such as N,N-dimethyl-4-aminopyridine (DMAP), N-hydroxybenzotriazole (HOBt), N-hydroxybenzotriazol (HOOBt), N-hydroxysuccinimide (HOSu) or 2-hydroxypyridine.

At that time, as in enzymatic peptide synthesis can usually be manipulated protective groups in chemical synthesis of reversible protection of reactive groups which do not react at about the synthesis of three preferred the traditional method of introducing protective groups: benzyloxycarbonyl (Z), tert-butoxycarbonyl (Boc) and 9-fertilitycare (Fmoc). In each case, identify the protective group on the amino group of the link extension chain. A detailed overview of the amino acid protective groups described in Mller, Methods der organischen Chemie, I. XV/1, S. 20-906, Thiemer Verlag, Stuttgart, 1974. The links used for the formation of the peptide chain can be subjected to interaction in solution, in suspension or by applying a method similar to that described in Merrifield, J. Amer.Chem.Soc. 85 (1963) 2149. Particularly preferred methods are those in which the peptides are collected sequentially, or by binding fragments using the method of the protective group Z, the First or Fmoc, and one reagent in said method Merrifield associated with an insoluble polymer carrier (hereinafter also referred to as a polymer). This usually leads to the fact that the peptide is assembled sequentially on the polymer carrier using the method of the protective groups are Boc or Fmoc, the growing peptide chain is covalently linked at the C-end with an insoluble polymer particles. This method gives an opportunity to separate the reagents and by-products by filtration and for this reason there is no need for recrystallization of the intermediate products.

Protected AB used solvents and should have a stable physical form, which facilitates filtration. The polymer must contain a functional group to which the first protected amino acid can be attached securely covalent bond. For this purpose, suitable wide variety of polymers, such as cellulose, polyvinyl alcohol, polymethacrylate, from sulphonated polystyrene, copolymer klimatisierung styrene/divinylbenzene (Merrifield resin), the resin 4-methylbenzhydrylamine (MBHA-resin), phenylacetamide-resin (Pam resin), n-benzyloxy-benzyl-alcohol-resin, benzhydrylamine-resin (BHA resin), 4-(hydroxymethyl) benzoyloxy-methyl/resin, resin Breipohl et al. (Tetrahedron Letters 28 (1987) 565; supplied by the company BACHEM), 4-(2,4-dimethoxyphenylacetic)-phenoxy-resin (supplied by company Novabiochem) or o-chlorotrityl-resin (supplied by company Biohellas).

For peptide synthesis in solution fit all solvents which are inert under the reaction conditions, especially water, N,N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), acetonitrile, dichloromethane (DCM), 1,4-dioxane, tetrahydrofuran (THF), N-methyl-2-pyrrolidone (NMP) and mixtures of these solvents. Peptide synthesis on polymeric carrier can be carried out in all inert organic solvents which do not nastojashie to the swelling of the polymers, such as DMF, DCM, NMP, acetonitrile and DMSO, and mixtures of these solvents. After completion of the synthesis the peptide otscheplaut from the polymer resin. Conditions for the removal of various types of polymers are disclosed in the literature. The most frequently used reactions of the cleavage reactions are catalyzed by acid and palladium, especially banding in liquid anhydrous hydrogen fluoride, anhydrous triftoratsetata, in diluted or concentrated triperoxonane acid, palladium-catalyzed cleavage in THF or mixtures of THF-DCM in the presence of a weak base, such as morpholine, or splitting in mixtures of acetic acid, dichloromethane and triptoreline. Depending on the selected protective groups, they may be withheld or similarly derived in terms of the removal of the polymer. Partial exemption from the protective groups of the peptide is useful when you need to carry out certain reactions to obtain derivatives. Peptides, dialkylamino the N-terminal nitrogen, can be obtained either by introducing the corresponding N,N-dialkylaminoalkyl in solution or on a polymeric carrier, or by reductive alkylation of peptide associated with the polymer in a mixture of DMF/1% acetic who lichnye diaminotoluene components, disclosed in this specification can be obtained from commercially available reagents, or synthesized from promyshlennosti reagents using known methods. For example, amino acids that make up the fragments with remains of R1and R2can be obtained in accordance with the work of E. Wsch, Houben Weyl, Meth. d. Org. Chemie, Bd. XV, 1, R. 306 following, Thieme Verlag Stuttgart 1974, and literary references in it. Peptides with bridges or lactam can be obtained by integrating the corresponding dipeptide units connected in bridge relationship with the lactam (R. Freidinger, J. Org. Chem. Soc. (1982) 104-109) in the peptide chain. Peptides with a thiazole, oxazole, thiazolin or oxazolidinedione dipeptide fragments can be obtained by integrating the corresponding dipeptide units (P. Jouin et al., Tetrahedron Letters (1992), 2807-2810; P. Wipf et al., Tetrahedron Letters (1992), 907-910; W. R. Tully, J. Med.Chem. (1991), 2065; Synthesis (1987), 235) in the peptide chain.

Compounds of the invention can be used for inhibition or other treatment of solid tumors (e.g., tumors of the lung, breast, colon, prostate, bladder, colon or endometrial tumors or hematological malignancies (such as leukemias, lymphomas) putea generally accepted in the pharmaceutical industry, preferably in Oncology using agents, including oral and parenteral routes, such as subcutaneous, intravenous, intramuscular and intraperitoneal administration. Connections can be entered individually or in the form of pharmaceutical compositions containing a compound of formula I together with a pharmaceutically acceptable carrier suitable for the desired route of administration. Such pharmaceutical compositions can be combined products, there may also contain other therapeutically active ingredients.

Dosage, administered to a mammal that contains an effective tumor-inhibiting amount of an active component, which depends on traditional factors, including biological activity of the specific compound, the route of administration, the age, health and weight of the recipient, the nature and status of symptoms, frequency of treatment, the introduction of other therapeutic drugs, as well as the desired effect. The usual daily dose is approximately 5-250 mg per 1 kg body weight by oral administration and approximately 1-100 mg per 1 mg of body weight at parenteral administration.

The new compounds can be administered in conventional solid or Gedaliah or solutions. They are made in the traditional way. For this purpose, the active substances can be processed with conventional pharmaceutical carriers, such as binder tablets, fillers, preservatives, disintegrators tablets, fluidity regulators, plasticizers, wetting agents, dispersing agents, emulsifiers, solvents, compositions prolonged action, antioxidants and/or propellants (see N.Sucker et al.: Pharmazeutische Technologie, Thieme-Verlag, Stuttgart, 1978). Forms for administration, obtained in this manner usually contain between 1 and 90% by weight of the active substance.

The following examples are intended to illustrate the invention. Proteinogenic amino acids are given in the examples in the reduction using a known three-letter code. Other designations are the following: TFA - triperoxonane acid, As acetic acid, Et is ethyl, Bu is butyl, Me is methyl, Bzl is benzyl.

A. General methods.

I. Peptides, stated in paragraph 1 of the claims, or synthesized by the classical method of synthesis in solution using standard Z - and Boc-method described above, or obtained by standard methods of solid-phase synthesis on an automated synthesizer model 431A supplied Phi is

a). Synthetic cycle for the method of the protective group Boc:

1. 30% triperoxonane acid in DCM - 1 x 3 min

2. 50% triperoxonane acid in DCM - 1 x 1 min

3. Flushing DCM - 5 x 1 min

4. 5% diisopropylethylamine in DCM - 1 x 1 min

5. 5% diisopropylethylamine in NMP - 1 x 1 min

6. Flushing NMP - 5 x 1 min

7. Adding pre-activated protected amino acids (activation with 1 equivalent of DCC and 1 equivalent of HOBt in NMP/DCM); the formation of the peptide bond (1st part) - 1 x 30 minutes

8. The addition of DMSO in the reaction mixture until then, until there is 20% DMSO by volume.

9. The formation of the peptide bond (2nd part) - 1 x 16 minutes

10. Adding 3.8 equivalent diisopropylethylamine in the reaction mixture.

11. The formation of the peptide bond (3rd part) - 1 x 7 minutes

12. Flushing DCM - 3 x 1 min

13. If the conversion is incomplete, repeat the stage of formation of the peptide bond (back to 5).

14. 10% acetic anhydride, 5% diisopropylethylamine in DCM - 1 x 2 minutes

15. 10% acetic anhydride in DCM - 1 x 4 minutes

16. Flushing DCM - 4 x 1 min

17. Return to l.

BOP-Cl and PyBrop used as reagents interaction napolitana or BOC-protected amino acid NCA (N-tert-butyloxycarbonyl-amino acid-N-carboxy-anhydrides), or Z-protected amino acid NCA (N-benzyloxycarbonyl-Millinocket-N-carboxy-anhydrides), respectively, is the most favorable for this type of reaction.

b). Synthetic cycle for the method of the protective Fmoc group:

1. Washing DMF - 1 x 1 min

2. 20% piperidine in DMF - 1 x 4 minutes

3. 20% piperidine in DMF - 1 x 16 minutes

4. Washing DMF - 5 x 1 min

5. Adding pre-activated amino acids (activating 1 equivalent of TBTU and 1.5 equivalents of DIPEA in DMF); the formation of the peptide bond - 1 x 61 minutes

6. Washing DMF - 3 x 1 min

7. If the transformation is not complete, repeat the formation of the peptide bond (back to 5).

8. 10% acetic anhydride in DMF - 1 x 8 minutes

9. Washing DMF - 3 x 1 min

10. Return to 2.

BOP-Cl and PyBrop used as reagents for the interaction of amino acid following the N-methyliminodiacetic. The reaction time is increased accordingly.

II. Reductive alkylation of the N-terminal nitrogen.

Peptide-polymer obtained in accordance with the stages of the Philadelphia Chapter and AB, released from the N-terminal protection (stage 2-4 in AB or 1-6 AIa) and then subjected to interaction with 3-fold molar excess and the negative test Kaiser, the polymer is washed several times with water, isopropanol, DMF and dichloromethane.

III. Treatment of the peptide-polymers obtained in accordance with the stages Ia and II.

Peptide-polymer is dried under reduced pressure and transferred into the reaction vessel of the apparatus TEFLON HF (supplied by the firm PENINSULA). After adding the acceptor, preferably anisole (1 ml/g of polymer), and in the case tylnej tryptophanase peptides, with the purpose of removal of inconformities group, preferably identicial (0.5 ml/g of polymer), carry out the condensation in hydrogen fluoride (10 ml/g of polymer), cooling the mixture of liquid N2. The mixture is left to warm to a temperature of 0oC and stirred at this temperature for 45 minutes Then the fluoride evaporated under reduced pressure and the residue is washed with ethyl acetate to remove the remaining acceptor. The peptide is extracted with 30% acetic acid and filtered, then the filtrate lyophilized.

IV. Treatment of the peptide-polymer obtained in stage IB and II.

Peptide-polymer is dried under reduced pressure and then subjected to one of the methods of removal are shown in table. 1, depending on the amino acid composition (Wade, Tregear, Howard Florey Fmoc Workshop Manual, Melbourne 1985).

A suspension of the peptide-polymer is p filtered and washed with TFA and DCM. The filtrate and the washing liquid concentrate and the peptide precipitated by adding diethyl ether. After cooling in an ice bath, the precipitate is filtered off, dissolved in 30% acetic acid and lyophilized.

V. When using the on-chlorotrityl-polymer (supplied by the company Biohellas), a suspension of the peptide-polymer in a mixture of acetic acid/triptoreline/dichloromethane (1:1:3.) stirred at room temperature for 1 hour the polymer was Then filtered off with suction and thoroughly washed with a solution for removal. The combined filtrates concentrated in vacuo and treated with water. The precipitated solid is removed by filtration or centrifugation, washed with diethyl ether and dried under reduced pressure.

VI. Purification and identification of peptides.

Cleaning is carried out by gel-chromatography (SEPHADEX G-10, 0-15/10% HOAc, SEPHADEX LH20/MeOH) with or without subsequent chromatography medium pressure (stationary phase: HD-SIL C-18, 2-45 μm, 100 mobile phase: gradient with A=0.1% of TFA/MeOH, B=0.1% of TFA/H2O).

The purity of the obtained products determine analytical ghvd (stationary phase: 100 2.1 mm VYDAC C-18, 5 l, 300 mobile phase: CH3CN/H2O gradient, sabotere the nd with the bombardment of heavy nuclei.

B. Specific examples.

Example 1 (SEQ ID NO: 1).

N,N-dimethyl-Val-Val-N-methyl-Val-Pro-Pro-Val-Phe-NH2< / BR>
1.98 g Fmoc-RINK-polymer (substitution and 0.46 mmole/g), the relevant portions from 0.84 mmole, is subjected to the interaction, as in stage IB, with the use of 1.26 mmole each of the following compounds:

Fmoc-Phe-OH Fmoc-N-methyl-Val-OH

Fmoc-Val-OH, Fmoc-Val-OH

Fmoc-Pro-OH, Fmoc-Val-OH

Fmoc-Pro-OH.

The amino acid following the N-methylaminoethanol associated with PyBrop as a reagent binding. After completion of the synthetic cycles peptide-polymer is subjected to a release from the N-terminal protection (stages 2-4 in AB) and then subjected to contact with an aqueous solution of formaldehyde, as in AII, and then dried under reduced pressure. The obtained polymer was subjected to TFA-cleavage, as in stage IV. The crude product (590 mg) purified by gel-filtration (SEPHADEX-LH-20). The output is 295 mg.

Example 2 (SEQ ID NO: 2).

< / BR>
4.11 g of polymer-Fmoc-Pro-p-alkoxybenzyl alcohol (substitution 0.73 mmole/g), corresponding portions 3 mmole, is subjected to the interaction, as in stage IB, using 4.5 mmole each of the following compounds:

Fmoc-N-MeVal-OH

Fmoc-Val-OH

Fmoc-Val-OH.

The amino acid following the N-methyl is when increasing the reaction time. After completion of the synthesis the peptide-polymer is subjected to a release from the N-terminal protection (stages 2-4 in AB) and then the interaction with the aqueous solution of formaldehyde, as in stage AII, and then dried under reduced pressure. Peptide-polymer thus obtained, is subjected to TFA-cleavage, as in stage IV. The crude product (750 mg) is used directly for further interaction. 100 mg of this compound is subjected to interaction with 45 mg of (S)-2-[1-amino-2-phenylethyl]thiazole and 230 mg of PyBop with the addition of 192 μl DIPEA in DMF at room temperature for 2 days. The reaction mixture was purified by gel-chromatography (SEPHADEX-LH-20, methanol) and the fraction of the product combine. Obtain 83 mg of product.

In accordance with examples 1 and 2 were obtained and can be obtained compounds are given in table. 2.

Examples to characterize using mass spectroscopy in respect of new synthesized compounds are given in table. 3.

In table. 4 shows the sequence of compounds obtained in accordance with examples 1 and 2.

The enumeration sequence.

(1) General information

(i) the Applicant

(A) BASF Aktiengesellschaft

(B) street: Carl-Bosch-Strasse 38

(C) city: Ludwigshafen

0

(ii) Title of invention: novel peptides, obtaining them and their use

(iii) Number of sequences: 57

(iv) a machine-Readable storage media:

(A) medium type: floppy, 3.5 inch, 2 x dual density

(B) computer: IBM AT-compatible processor 80286

(C) operating system: MS-DOS version 5.0

(D) software: WordPerfect version 5.1

(2) the Information about the sequence SEQ ID NO: 1:

(i) sequence Characteristics:

(A) length: 9 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 1:

< / BR>
(2) the Information about the sequence SEQ ID NO: 2:

(i) sequence Characteristics:

(A) length: 5 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 2:

< / BR>
(2) the Information about the sequence SEQ ID NO: 3:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 3:

< / BR>
(2) the Information about the sequence SEQ ID NO: 4:

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 4:

< / BR>
(2) the Information about the sequence SEQ ID NO: 5:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 5:

< / BR>
(2) the Information about the sequence SEQ ID NO: 6:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 6:

< / BR>
(2) the Information about the sequence SEQ ID NO: 7:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 7:

< / BR>
(2) the Information about the sequence SEQ ID NO: 8:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 8:

< / BR>
(2) the Information about the sequence SEQ ID NO: 9:

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 9:

< / BR>
(2) the Information about the sequence SEQ ID NO: 10:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 10:

< / BR>
(2) the Information about the sequence SEQ ID NO: 11:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 11:

< / BR>
(2) the Information about the sequence SEQ ID NO: 12:

(i) sequence Characteristics:

(A) length: 6 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 12:

< / BR>
(2) the Information about the sequence SEQ ID NO: 13:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 13:

< / BR>
(2) the Information about the sequence SEQ ID NO: 1 (D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 14:

< / BR>
(2) the Information about the sequence SEQ ID NO: 15:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 15:

< / BR>
(2) the Information about the sequence SEQ ID NO: 16:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 16:

< / BR>
(2) the Information about the sequence SEQ ID NO: 17:

(i) sequence Characteristics:

(A) length: 6 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 17:

< / BR>
(2) the Information about the sequence SEQ ID NO: 18:

(i) sequence Characteristics:

(A) length: 6 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 18:

< / BR>
(2) the Information about the sequence SEQ ID NO: 1 (D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 19:

< / BR>
(2) the Information about the sequence SEQ ID NO: 20:

(i) sequence Characteristics:

(A) length: 6 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 20:

< / BR>
(2) the Information about the sequence SEQ ID NO: 21:

(i) sequence Characteristics:

(A) length: 6 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 21:

< / BR>
(2) the Information about the sequence SEQ ID NO: 22:

(i) sequence Characteristics:

(A) length: 6 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 22:

< / BR>
(2) the Information about the sequence SEQ ID NO: 23:

(i) sequence Characteristics:

(A) length: 4 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 23:

< / BR>
(2) the Information about the sequence SEQ ID NOR> (D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 24:

< / BR>
(2) the Information about the sequence SEQ ID NO: 25:

(i) sequence Characteristics:

(A) length: 4 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 25:

< / BR>
(2) the Information about the sequence SEQ ID NO: 26:

(i) sequence Characteristics:

(A) length: 4 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 26:

< / BR>
(2) the information about the sequence SEQ ID NO: 27:

(i) sequence Characteristics:

(A) length: 5 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 27:

< / BR>
(2) the Information about the sequence SEQ ID NO: 28:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 28:

< / BR>
(2) the Information Is(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 29:

< / BR>
(2) the Information about the sequence SEQ ID NO: 30:

(i) sequence Characteristics:

(A) length: 6 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 30:

< / BR>
(2) the Information about the sequence SEQ ID NO: 31:

(i) sequence Characteristics:

(A) length: 4 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 31:

< / BR>
(2) the information about the sequence SEQ ID NO: 32:

(i) sequence Characteristics:

(A) length: 4 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 32:

< / BR>
(2) the Information about the sequence SEQ ID NO: 33:

(i) sequence Characteristics:

(A) length: 6 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 33:

< / BR>
(2) the Information is BR> (B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 34:

< / BR>
(2) the Information about the sequence SEQ ID NO: 35:

(i) sequence Characteristics:

(A) length: 5 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 35:

< / BR>
(2) the Information about the sequence SEQ ID NO: 36:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 36:

< / BR>
(2) the information about the sequence SEQ ID NO: 37:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 37:

< / BR>
(2) the Information about the sequence SEQ ID NO: 38:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 38:
(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 39:

< / BR>
(2) the Information about the sequence SEQ ID NO: 40:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 40:

< / BR>
(2) the Information about the sequence SEQ ID NO: 41:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 41:

< / BR>
(2) the Information about the sequence SEQ ID NO: 42:

(i) sequence Characteristics:

(A) length: 6 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 42:

< / BR>
(2) the Information about the sequence SEQ ID NO: 43:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: (A) length: 6 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 44:

< / BR>
(2) the Information about the sequence SEQ ID NO: 45:

(i) sequence Characteristics:

(A) length: 5 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 45:

< / BR>
(2) the Information about the sequence SEQ ID NO: 46:

(i) sequence Characteristics:

(A) length: 6 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 46:

< / BR>
(2) the Information about the sequence SEQ ID NO: 47:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 47:

< / BR>
(2) the Information about the sequence SEQ ID NO: 48:

(i) sequence Characteristics:

(A) length: 6 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO:(A) length: 6 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 49:

< / BR>
(2) the Information about the sequence SEQ ID NO: 50:

(i) sequence Characteristics:

(A) length: 6 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 50:

< / BR>
(2) the Information about the sequence SEQ ID NO: 51:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 51:

< / BR>
(2) the Information about the sequence SEQ ID NO: 52:

(i) sequence Characteristics:

(A) length: 6 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 52:

< / BR>
(2) the Information about the sequence SEQ ID NO: 53:

(i) sequence Characteristics:

(A) length: 5 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO (A) length: 5 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 54:

< / BR>
(2) the Information about the sequence SEQ ID NO: 55:

(i) sequence Characteristics:

(A) length: 6 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 55:

< / BR>
(2) the Information about the sequence SEQ ID NO: 56:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 56:

< / BR>
(2) the Information about the sequence SEQ ID NO: 57:

(i) sequence Characteristics:

(A) length: 7 amino acids

(B) type: amino acid

(D) topology: linear

(ii) Molecular type: peptide

(xi) sequence Description: SEQ ID NO: 57:

< / BR>
Abbreviations have the meanings given in Fig. 42-71.

The end-NH2means that the C-terminal amino acid is in its amide form.

Compounds of the invention can be tested for antitumor activity of Tr is ü measured using standard methods for lines of adhesive cells, such as using tetrazolium test for algae (MTT). Details of this analysis are published (Alley, MS et al. , Cancer Research, 48: 589-601, 1988). Exponentially growing cultures of tumor cells, such as cells HT-29 carcinoma of the colon or LX-1 lung tumor, is used in order to obtain a cell culture in the blade. Cells were seeded at a concentration of 5,000 - 20,000 cells per well in 96-well plates (150 μl medium) and grown over night at 37 ° oC. Add compound with a 10-multiple dilutions ranging from 10-4M to 10-10M. the cells are Then incubated for 48 hours To determine the number of viable cells in each well add the dye MTT (50 μl of 3 mg/ml 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium in saline solution). This mixture is incubated at a temperature of 37oC for 5 h, after which each well add 50 μl of 25% aqueous sodium dodecyl sulfate (SDS), pH 2. After incubation overnight spectral absorption capacity of each well at 550 nm is recorded using a tablet reader enzyme-linked immunosorbent assay. Calculate values for the average standard deviation +/- from data from mancentral of the test compound, which gives About/To equal 50% of the resultant inhibition of growth, denoted as IC50(inhibiting growth by 50% concentration).

B. Methodology in vivo.

Compounds of the invention can be further tested in any of a variety of preclinical tests for activity in vivo, which is an indicator of clinical suitability. These tests are carried out on hairless mice, which transplanted ("xenotransplantation") tumor tissue, preferably of human origin, which is well known in the art. Compound evaluated for their antitumor efficacy after administration xenotransplantation mice. More specifically, human tumors grown in hairless mice (Nude mice with a mutation in the gene nude), transplanted new recipient animals with tumor fragments, which have a size of about 50 mg. a Day of transplantation designated as day 0. After 6-10 days, mice treated with the test compound, administered intravenously or intraperitoneally in the form of injections, in groups of 5-10 mice per dose. Join prescribed daily for 5 days, 10 days or 15 days at a dosage of 10-100 mg/kg of body weight. The diameter of the swollen the calipers of Vernier, and formulas

(length x width2)/2 + mg tumor mass

The average weight of tumors calculated for each treatment group, and the values Of/To determine in respect of each group relative to the untreated control tumors.

The new compounds of the invention exhibit good activity in vitro in the above systems analysis and antitumor activity in the above-mentioned system in vivo.

Listed below are the data on the characteristic properties of the obtained compounds.

In particular, provides information about the mass spectroscopy of peptides (see table. 3). Using the methodology As that disclosed in the application materials, these compounds were analyzed for anti-tumor activity. The results of this analysis are presented in table. 5. Next, we studied the cytotoxicity of these peptides using crystal violet staining (see tab. 6). Some of the compounds have been tested in preclinical analysis to determine the antitumor activity in vivo. In table. 8 are given the appropriate information.

Salts of the claimed compounds can be obtained by treating the peptides equivalent amount of acid and the concentration of sediment. In table. 8 shows the data availability antitumor activity of the peptides according to the invention. In addition, they contain reliable data, confirming the suitability of the inventive method for the treatment of cancer in mammals. Since the new compounds in the in vivo tests did not show toxicity, they can be considered low-toxic or slightly toxic.

The compounds of this invention can be analyzed for antitumor activity using conventional techniques, for example, shown earlier.

A. the in vitro Method described above.

The results of the analysis of antitumor activity of the compounds are given in table. 5.

Further compounds according to the invention were tested for antitumor activity using quantitative analysis of crystal violet on cytotoxicity.

This analysis was carried out in accordance with the method described Flick N. and Gifford, G. E., J. Immunol. Meth. 68, 167-175 (1984). Proteins viable monolayer cells were stained after exposure to cytotoxic drug and colorimetrically was measured quantity. Test cells (SK-1, clone carcinoma) was placed in a 96-aerated well microtiter plates at a density 2-3103cells/well and were cultured under standard conditions stump within 1 day. Then cells were exposed to the effect of the test compounds at several concentrations. Controls were incubated in the medium separately. After further incubation for 72 h, culture medium was mixed by shaking and added 58 μl colored crystal violet in each well. After staining for 20 min colored solution was decanted and the plates were washed in a strong jet of water up until all unbound dye is removed.

The remaining insoluble crystalline dye was dissolved by adding 100 μl of a solution containing 50% ethanol and 0.1% acetic acid to each well. The monolayer in each well was determined using a microtiter ELISA plates counter at 540 nm (Titertec Multiscan, Flow. Lab. Mechenheim). The concentration of test compound which gives T/C 50% inhibition of growth was identified as ID50. The results of the analysis of the cytotoxic activity of the described compounds are given in table. 6.

Methods in vivo.

Further compounds according to the invention was tested in pre-clinical analysis to determine activity in vivo, which indicates their clinical suitability. Analysis was performed on hairless mice, which were t is all right known from the art. The tested compounds were evaluated for their antitumor efficacy after administration to mice bearing xenograft. More specifically, tumor human breast (MX-1), which were grown in hairless mice (Nude mice with a mutation in the gene nude) transplanted into new recipient mice. The day of transplantation was defined as day 0. After 6-10 days, the mice were treated with the test compounds, which were injected intravenously with each dose used groups of 5-10 mice. Compounds were given every day for 3 weeks, at a dose of 1-100 mg/kg of body weight. The diameter of the tumor and body weight were measured twice a week. Tumor size was determined using the diameter measured by the compass of Vernier, and formulas

(length x width2)/2 = mm3tumor volume (Fig. 72).

The value of the tumor mass was calculated for each treated group and the T/S value was determined for each of the respective untreated control tumors.

In Fig. 72 Dana antitumor activity on the example of breast cancer (compound of example 1).

The second example shows mice that were treated with compound of example 234, 25 mg/kg three times per week for three weeks the Shi died. Novarossi tumors was observed in treated mice.

The tumor regression was observed at doses that were not toxic to horses (Fig. 73).

In Fig. 73 given the antitumor activity of the example compound 234.

A comparative study of compound of example 234 and its hydrochloride in vivo and in vitro. Cytotoxicity in vitro is shown in table. 7.

Activity in vivo against MX-l, human carcinomas of the breast xenotransplantion ('s.c.) hairless mice.

In Fig. 74 given the antitumor activity of the compounds according to the invention in vitro against MX-1 human breast carcinoma.

Free amides and their hydrochloride showed the same biological activity.

Similar compounds were tested in vivo.

Other compounds were also investigated on the model MX-l, as described below. Compounds were studied i.v.

Compounds that have good activity on the MX-1 model were also tested additional models in vivo. The effect on tumor growth of the described compounds tested in vivo, are given in table. 8.

These data show that the new compounds possess good properties of inhibitors of p is l, cycloalkyl, foralkyl, aminosulfonyl, which can be substituted by alkyl; hydroxy or NR3R4where each R3and R4can be either hydrogen or alkyl;

R2is hydrogen, alkyl, foralkyl, cycloalkyl;

R1-N-R2together may denote a 5-membered heterocycle which can be unsubstituted or substituted by one or more substituents, represents an alkyl group;

And the residue was felled, isoleucyl, laitila, allasalueella, aminoisobutyrate, 3-tert-butylamine, 2-tert-butylphenyl, 3-cyclohexylethyl, 2-ethylglycine, 2-cyclohexylglycine, norleucine, Norvaline;

In - N-alkyl-poured, -Norvaline-Lalla-isoleucyl, -2-tert-butylphenyl, 3-tert-butylamine, 3-cyclohexylamine, i.e. phenylalanyl or-2-cyclohexylglycine;

D, E, F and G independently are selected from the group consisting of prolyl residues, homepolish, hydroxyproline, thiazolidine-4-carbonyl, 1-aminopentyl-1-carbonyl, thronged, 2-tert-butylglycol, isoleucyl, laitila, 3-cyclohexylamine, i.e. phenylalanyl, N-methylphenylamine, tetrahydroisoquinoline-2-carbonyl, 3-thiazolidinone, 3-thienylene, histidine, 1-aminoindane-1-carbonyl, arginine, 3-pyridylethyl, 3-tert-bout is CH2-cyclohexyl or arylalkyl;

E and F together may designate

< / BR>
< / BR>
< / BR>
where Y is hydrogen or lower alkyl;

Z denotes hydrogen or lower alkyl;

n = 1, 2, or 3;

V is oxygen or sulphur;

M is hydrogen, lower alkyl, arylalkyl, cyclohexyl, or-CH2-cyclohexyl;

Q is hydrogen;

R is hydrogen or lower alkyl;

or R and Q can together form a bond;

U is hydrogen, lower alkyl, phenyl or cycloalkyl;

W is hydrogen, lower alkyl or phenyl;

u, v and w are independently 0 or 1;

K - hydroxy, alkoxy, phenoxy, benzyloxy or substituted or unsubstituted amino group,

and their salts with physiologically compatible acids.

2. The peptides of formula I under item 1, wherein R1- N - R2denotes a 5-membered heterocycle of the formula

< / BR>
< / BR>
which may be unsubstituted or substituted by one or more substituents, represents an alkyl group.

3. The peptides of formula I under item 1, characterized in that the amino group of the formula

R5- N - R6,

in which R5is hydrogen, lower alkyl, lower alkoxy, benzyl;

R6is hydrogen, lower alkyl, phenyl, C5-cycloalkyl, benzyl, unsubstituted or is/SUP> - the five-membered heteroaryl, where R7is hydrogen, benzyl, and heteroaryl selected from the group; thiazole, thiophene, oxazole.

4. Derivative of the peptide of General formula I on p. 1, representing the peptides of formula I, where u, v and w are equal to 0 and is not hydroxy, benzyloxy, phenoxy or alkoxygroup.

5. Derivative of the peptide of General formula I on p. 1, representing the peptides of formula I, where u and v are equal to 0 and does not denote a hydroxy - or alkoxygroup.

6. Derivative of the peptide of General formula I on p. 1, representing the peptides of formula I, where u, v and w are equal to 1 and denotes hydroxy-, alkoxy-, phenoxy or benzyloxy.

7. Derivative of the peptide of General formula I on p. 1, representing the peptides of formula I, where u and v are equal to 1, w is 0 and K denotes a hydroxy-, alkoxy-, phenoxy or benzyloxy.

8. Derivative of the peptide of General formula I on p. 1, representing the peptides of formula I, where u is 1, v and w are equal to 0 and K denotes a hydroxy-, alkoxy-, phenoxy or benzyloxy.

9. Derivative of the peptide of General formula I under item 1, with the tumor-inhibitory activity.

10. Pharmaceutical composition having the tumor-inhibitory activity, including AK is I on p. 1 in an effective amount.

 

Same patents:

The invention relates to new peptides with organizaitnal activity with high biological activity of the same type as the natural compound HRV, but with a shorter amino acid chain

The invention relates to new biologically active compounds, specifically, the peptides of General formula: Trp-X-Gly-Gly-Asp-R, where X is the residue of the hydroxyl - containing amino acids L-or D-configuration, R-Ala-Ser-Gly-Glu or Ala-Ser-Gly or Ala-Ser, or Ala; and their pharmaceutically acceptable salts having anti-stress, anticonvulsant and neuroprotective action

The invention relates to peptides of formula (I): X - A1- A2- Thr - Ala - Val - Gly - His - Leu - psi - A9- Q, where X represents a hydrogen, a simple relationship linking the alpha-amino group of A1with gamma-carboxyl part 3-propionyloxy part of A2if A2is Glu[-], or a group of the formula R1CO-, where R1selected from the group comprising: hydrogen, C1- C10- alkyl, or phenyl C1- C10- alkyl, A1is a D - or L-amino acid residue selected from the group consisting of: Phe, p - Hl - Phe, pGlu, Nal, Pal, Tpi, unsubstituted Trp or Trp substituted in the benzene ring by one or more substituents from the group comprising C1- C3- alkyl, or A1represents a peptide bond linking the acyl part of R1CO with alpha aminocyclo A2if A2represents Gln, Glu/-/ Glu (Y) or His, where /-/ is a simple relationship linking the gamma-carboxyl group of A2with the alpha-amino group of A1if A2is Glu, where X represents a simple bond, Y represents - or SIG5where R5is hydrogen, C1- C3- alkyl or phenyl; Leu - psi - is a reduced form Lой adjacent A9- balance is pseudopeptides communication; A9is a TAS, Ista, or DМТас; and Q represents NH2or CQ1where Q1is hydrogen, and pharmaceutically acceptable acids or salts, and pharmaceutical compositions, which has antagonistic activity against bombezin and to a method of treating cancer in mammals on the basis of the peptides of formula (I)

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The invention relates to the field of natural physiologically active peptides, specifically to an improved method for producing a peptide-sleep formula I:

TrpAlaGlyGlyAspAlaSerGlyGlu

Peptide-sleep has anti-stress [1], protivoallergennoy [2], antimetastatic [3] and other types of biological activity

The invention relates to new peptides with high biological activity of the same type, which is inherent in the known natural compound BPC, but with a shorter amino acid chain

The invention relates to medicine, namely to pharmacology, and more specifically to methods of producing biologically active substances, and may find application in the clinic, veterinary, as well as in experimental studies

The invention relates to new peptides bradykinin-antagonistic action, and method of production thereof

The invention relates to new chemical substances that have valuable biological properties, and more particularly to a derivative of a peptide of formula (I):

AA1-AA2-AA3-AA4-AA5-AA6(I)

where AA1group D - or L-N-thioxanthine, D - or L-N-centerlized, D-5H-dibenzo(a, d)cycloheptanol, L - or D-10,11-dihydro-5H-dibenzo(a, d)(cyclohepten-5-yl)glycine or L - or D--amino-10,11-dihydro-5H-dibenzo(a, d)cyclohepten-5-acetic acid, the amino acids may have a protective group;

AA2leucine, arginine, ornithine, or glutamic acid;

AA3aspartic acid, N-metilparabena acid;

AA4isoleucine, phenylalanine;

AA5isoleucine, N-methylisoleucine;

AA6tryptophan, N-formylthiophene

or their pharmaceutically acceptable salts

The invention relates to therapeutic peptides suitable for the treatment of benign or malignant tumors and disorders of the gastrointestinal tract

The invention relates to new biologically active compounds, specifically, the peptides of General formula: Trp-X-Gly-Gly-Asp-R, where X is the residue of the hydroxyl - containing amino acids L-or D-configuration, R-Ala-Ser-Gly-Glu or Ala-Ser-Gly or Ala-Ser, or Ala; and their pharmaceutically acceptable salts having anti-stress, anticonvulsant and neuroprotective action

The invention relates to peptides of formula (I): X - A1- A2- Thr - Ala - Val - Gly - His - Leu - psi - A9- Q, where X represents a hydrogen, a simple relationship linking the alpha-amino group of A1with gamma-carboxyl part 3-propionyloxy part of A2if A2is Glu[-], or a group of the formula R1CO-, where R1selected from the group comprising: hydrogen, C1- C10- alkyl, or phenyl C1- C10- alkyl, A1is a D - or L-amino acid residue selected from the group consisting of: Phe, p - Hl - Phe, pGlu, Nal, Pal, Tpi, unsubstituted Trp or Trp substituted in the benzene ring by one or more substituents from the group comprising C1- C3- alkyl, or A1represents a peptide bond linking the acyl part of R1CO with alpha aminocyclo A2if A2represents Gln, Glu/-/ Glu (Y) or His, where /-/ is a simple relationship linking the gamma-carboxyl group of A2with the alpha-amino group of A1if A2is Glu, where X represents a simple bond, Y represents - or SIG5where R5is hydrogen, C1- C3- alkyl or phenyl; Leu - psi - is a reduced form Lой adjacent A9- balance is pseudopeptides communication; A9is a TAS, Ista, or DМТас; and Q represents NH2or CQ1where Q1is hydrogen, and pharmaceutically acceptable acids or salts, and pharmaceutical compositions, which has antagonistic activity against bombezin and to a method of treating cancer in mammals on the basis of the peptides of formula (I)

The invention relates to new peptides with high biological activity of the same type, which is inherent in the known natural compound BPC, but with a shorter amino acid chain

The invention relates to medicine, namely to pharmacology, and more specifically to methods of producing biologically active substances, and may find application in the clinic, veterinary, as well as in experimental studies

The invention relates to medicine, namely to methods of producing biologically active substances with immunoregulatory properties, and may find application in medicine,veterinary medicine and experimental biochemistry

The invention relates to medicine, namely to methods of producing biologically active substances with immunoregulatory properties, and may find application in medicine, veterinary medicine and experimental biochemistry

The invention relates to new chemical substances that have valuable biological properties, and more particularly to a derivative of a peptide of formula (I):

AA1-AA2-AA3-AA4-AA5-AA6(I)

where AA1group D - or L-N-thioxanthine, D - or L-N-centerlized, D-5H-dibenzo(a, d)cycloheptanol, L - or D-10,11-dihydro-5H-dibenzo(a, d)(cyclohepten-5-yl)glycine or L - or D--amino-10,11-dihydro-5H-dibenzo(a, d)cyclohepten-5-acetic acid, the amino acids may have a protective group;

AA2leucine, arginine, ornithine, or glutamic acid;

AA3aspartic acid, N-metilparabena acid;

AA4isoleucine, phenylalanine;

AA5isoleucine, N-methylisoleucine;

AA6tryptophan, N-formylthiophene

or their pharmaceutically acceptable salts

The invention relates to medicine, in particular to surgery, and can be used for the treatment of patients with hemorrhagic shock

The invention relates to medicine, namely to means for reducing pathological vascular permeability to proteins

FIELD: medicine, cardiology.

SUBSTANCE: the suggested method should be performed at the background of medicinal therapy with preparations out of statins group, tevetene, polyoxidonium and conducting seances of plasmapheresis by removing 800 ml plasma twice weekly with N 5 due to additional intramuscular injection of immunophan 0.005%-1.0 with N 10 and fluimucyl 300 mg intravenously daily with N 5-10, total course of therapy lasts for 2 mo. The method provides modulation of leukocytic functional activity, moreover, due to altered cytokine profile and, thus, through disintegration of protein-lipid complexes participating in the development of atherosclerotic platelets.

EFFECT: higher efficiency of therapy.

3 ex

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