Derivatives nonapeptides or their salts with pharmaceutically acceptable acids, method for their production and pharmaceutical composition

 

(57) Abstract:

Usage: in medicine, as having antagonistic activity against bombezin. The essence of the invention: derivatives of nonapeptides General formula I: X - A1- A2- Trp - Ala - Val - Gly - His - Leu - psi TpiQ, where Q = OMe, NH2X = hydrogen, acetyl, CONH2; A1= (D or L)Phe, (D - or L-)Tpi, D-p-Glu; A2= Gln, Glu(OMe), Glu(MeNH), Glu, His(Bz) or formula II: D - Tpi - A2- Trp - Ala - Val - Gly - His - Leu-psi-A9NH2where A2= Gln, Glu(OMe), His(Bz); A9= Leu, Ple, Trp, Trp(For) or their salts with pharmaceutically acceptable acids, obtained by stepwise extension of the peptide chain, which is carried out by protecting the amino group, in-position, and the end carboxypropyl covalently bind with a synthetic polymer carrier used for this purpose, after which protecting group unhooking, carboxypropyl subsequent amino acids to restore education group which is then associated with the above-amino group, subjected to the pre-release, then aminoamides group of the second amino acid will unlock and carboxypropyl subsequent amino acids are then associated with the above-mentioned released-amino group of a second is denaut other amino acids, from the resulting peptidyl-polymer otscheplaut media and, if necessary, the protective group of the obtained peptides if necessary acelerou and the target product is isolated in free form or in the form of a salt with a pharmaceutically acceptable acid, the pharmaceutical composition comprises the active ingredient (compound of formula (I) in an effective amount and a pharmaceutically acceptable carrier. The proposed method allows you to gain valuable new connections that are of interest for the treatment of malignant diseases in warm-blooded animals and humans. 5 C. and 14 C.p., four crystals, 2 tab.

The present invention is made with Government support in the form of subsidies N CA 40077 provided by the National cancer Institute (national institutes of health). The U.S. government has certain rights in this application.

The present invention aims to obtain previously unknown peptides, which have an impact on the growth of malignant tumors in humans. More specifically, the present invention concerns anteromesial, which are a [8-9- pseudo] -nonapeptide containing D - or L - tryptophan or tryptophan analogue 2,3,4,9-tetrahydro-volume C, which have antagonistic properties against bombesin or Bambisanani peptides, salts thereof, and pharmaceutical compositions and methods of use in relation to these peptides.

The present invention relates to a polypeptide compounds that possess antagonistic properties against bombesin or Bambisanani peptides, such as gastronomically peptide, neuromedin C and similar substances, then nazyvaemymi anteromedially properties, and which are of interest, for example, for the treatment of malignant diseases in warm-blooded animals such as man. The invention relates to previously unknown polypeptide compounds and methods for their production, previously unknown pharmaceutical compositions containing the aforementioned peptide compounds, and methods of preparation of pharmaceutical preparations containing such, to obtain antivomiting effect in warm-blooded animals and humans.

The bombezin is tetradecapeptide amide, which was first isolated from the skin of the frog species Bombinabombina (Anastasi, Erspamer and Bucci, Experientia, 1971, 27, 166). It is known that the bombezin is a potent mitogen in respect of amylase of pancreatic acini from Guinea pigs (Jensen, Jones, Folkers and Gardner, Nature, 1984, 309, 61). It is known that bambusoideae peptides are produced and are secreted human melolonthinae pulmonary malignant cells (Moody, Pert, Gazdar, Carney and Minna, Science, 1981, 214, 1246) that exogenously introduced bambusoideae peptides can stimulate the growth of human small cell lung cancer cells in vitro (Carney, Cuttia, Moody and Minna, Cancer Research, 1987, 47, 821), and that a monoclonal antibody specific against the C-terminal region of bombezin and gastronomiebedarf peptide can block the binding gastronomiebedarf peptide, with its receptors and inhibit growth of human small cell lung cancer cells in vitro and in vivo (Cuttita, Carnce, Mulshine, Moody, Fedorko, Fischler and Minna, Nature, 1985, 316, 823).

Gastronomically peptide, which has bambusoideae properties, is a widely distributed peptide amide containing 27 amino acids isolated from porcine intestine (Mc Donald, Jornvall, Nilsson, Vagn Ghatei, Bloom and Mutt, Biochem. Biophys. Res. Commun., 1979, 90, 227), whose C-terminal amino acid sequence is almost similar to that of the bombezin. Neuromedin C represents Decapeptide amide, the structure of which is identichniye, which was separated from the dog small intestine (Reeve, Walsh, Chew, Clark, Hawke and Shively, J. Biol. Chem., 1983, 258, 5582). Gastronomically peptide stimulates the emergence of a variety of biological reactions, including the release of gastrin in the systemic circulation. It also functions as a growth factor in fibroblasts mouse line T and in small-cell lung malignant cell. Thus, it turns out that gastronomically peptide plays a direct pathophysiologically role in the development of small cell lung malignant cells that occur by the mechanism of autocrine growth.

Patterns of bombezin, neuromedin C and carboxyl-terminal nonapeptide for gastroesophageal peptide is shown below:

bombesin pGlu-Gln-Arg-Leu-Gly-Asn-Gln-Trp-Ala-Val-Gly-His - Leu-Met-NH2< / BR>
neuromedin C H-Gly-Asn-His-Trp-Ala-Val-Gly-His-Leu-Met-NH2< / BR>
C-terminal nonapeptide-Asn-His-Trp-Ala-Val-Gly-His-Leu-Met-NH2< / BR>
gastroesophageal peptide

Search other antibiotic Bambisanani peptides led to the isolation of litorina, nonapeptide (pGlu-Gln-Trp-Ala-Val-Gly-His-Leu-Met-NH2located in the skin of frogs from Papua, New Guinea, which, as it turns out, is the most potent bombesin (Yasukara et.al., Chem. Pharm amino acid residues, taken from 6-14 at bombezin, has the full range babusikova activity.

It is known now several types of anteromesial. Substance P(Arg - Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2), which in its amino acid sequence is characterized by the presence of small homology with bombezin, did not inhibit the binding of bombezin and Bambisanani peptides; however, analogs of substance P, modified by the replacement of several L - amino acids to D-amino acids such as D-Arg1D-Pro2, D-Trp7,9, Leu11) - substance P and (D-Arg1, D-Phe5, D-Trp7,9, Leu11) - substance P (Moody et al., Fed. Proceedings, 1987, 46, 2201) block, as it is established, the secretion of bombezin in pancreatic acinar cells and inhibit the effects associated with the promotion of the growth of bombezin, in the cell line Swiss T. It turned out that two types antonomasia isolated from bombezin, for example, (D-Phe6, D-Phe12- bombezin and (Leu13-psi-Leu14- bombezin (Coy et al, J. Biol. Chem., 1988, 263, 5056 and peptides 1989, 10, 587), are in vitro and in vivo potent inhibitors of the reaction on the bombezin.

Another type antonomasia identified by Heimbrock and others (Heimbrook et al, J. Biol. Chem., 1989, 264, 11258) represents N-acetyl formed scenovisionmoliere peptide, (with the number of members in the chain 20 - 27) is a remote C-terminal meinenemy balance. Recently Coy (Coy, J. Biol. Chem 1989, 264, 14691) reported that some antiobesity short circuit, based on the Littorina sequence, such as (D-Phe6, Leu13-psi-Phe14-bombezin-(6 - 14) and (D-Phe6, Leu13-[psi-Leu14)-bombezin-(6 - 14), detect significantly higher selenocystine than their corresponding original peptide (Leu13-psi-Leu14-bombezin.

The present invention relates to previously unknown polypeptides that are potent anteromedially, methods for their preparation, pharmaceutical compositions comprising the above-mentioned polypeptides and their use as pharmaceutically active compounds.

More specifically, the present invention concerns pseudopeptides comprising the polypeptide component of the formula I

X-A1-A2A3-A4-A5-A6-A7-A8-psi-A9-Q

where

Q is a group NH2or OQ1where Q1is hydrogen, C1-10-alkyl, phenyl or C7-10-fenilalanina group;

X is hydrogen or a single bond connecting the alpha amino group of residue A1with the group with the formula:

R1CO-,

in the case where R1choose from the groups consisting of:

a) hydrogen, C1-10-alkyl, phenyl or C7-10-phenylalaninol group;

b)

< / BR>
where

R2is hydrogen, C1-10-alkyl, phenyl or C1-10-alkyl, phenyl or C7-10-fenilalanina group; R3is hydrogen or C1-10is an alkyl group and

in R4-O-

where

R4- C1-10-alkyl, phenyl or C7-10-fenilalanina group;

A1-D-, L - or DL-pGlu, Nal, Phe, Thi, Tyr, Tpi, Hca, Hpp, Mpp, Trp or Trp group substituted in the benzene ring by one or more members selected from the group comprising halogen, NO2, NH2, OH, C1-3is an alkyl group, and C1-3-alkoxygroup, where the halogen is fluorine, chlorine and bromine;

A2-Ash, Dpa, Gln, His, MeHis, His (Bz), His (Z) or a group with the formula Asp (V), Glu [-] and Glu (V) where: V - group OR5or , where R5is hydrogen, C1-3is an alkyl or phenyl group, R6is hydrogen, C1-3is an alkyl group or a group-NHCONH2and [-] - single link connecting the lateral carboxyl group to the alpha-amino group of education A1where X is a single bond;

A3- Nal, Pal, Tpi, Trp, MeTrp, Trp (For) or group Tr is Yong, NO2, NH2, OH, C1-3is an alkyl group, and C1-3-alkoxygroup, where the halogen is fluorine, chlorine and bromine;

A4- Ala, MeAla or Gln;

A5- Val or MeVal;

A6- Gly, Phe or D-Ala;

A7Is His, MeHis, His(Bz), His(Z), Lys (Z) or Pal;

A8-restored isostere from Leu or Phe;

A9-Leu, Phe, Tpi, Trp or Trp group substituted in the benzene ring by one or more members selected from the group comprising halogen, NO2, NH2, OH, C1-3is an alkyl group, and C1-3-alkoxygroup, where the halogen is fluorine, chlorine and bromine, with a prerequisite that the group A1or A9represents the D-, L - or DL-Tpi group, and salts thereof with pharmaceutically acceptable acids.

Preferred polypeptides with formula I are polypeptides that have A1is selected from the group comprising Mpp, D-Phe, L-or D-Tpi, D-Trp or Trp group substituted in the benzene ring by one or more members selected from the group comprising halogen, NO2, NH2, OH, C1-3-alkoxygroup, where the halogen is fluorine, chlorine and bromine, A9- D -, L - or DL-amino acid residue selected from the group comprising (Leu) Phe, Tpi, Trp or Trp(For).

Reduction of individual amino acid residues are based on a standard called amino acids, for example, (Trp - Triptrap, Gln is glutamine, His is histidine, Ala - Ala, Val-valine, Gly is glycine, Leu is leucine, Phe is phenylalanine. If the amino acid residue isomeric forms refers to the amino acid in L-form, unless a different form is not mentioned specifically, in this case, before the designation of the amino acids for the different forms appear denote a D - or DL.

In the present invention to denote an unusual amino acids taken the following abbreviations:

Dpa - 2,3-diaminopropionic acid,

Nal-3-(2-naphthyl)-alanine,

Thi - -2'-titillans,

Tpi-2,3,4,9-tetrahydro-IH-pyrido-(3,4-b) -indole-3-carboxylic acid.

Peptide sequences are written with the condition that the N-terminal amino acid is on the left side and the C-terminal acid is on the right side.

Hca - hydrocinnamic acid,

Hna-3-hydroxy-2-naphthoic acid,

Hpp-3-(4-hydroxyphenyl)propionic acid,

Mpp - 3-(4-methoxyphenyl)propionic acid,

Paa - phenylacetic acid,

Other abbreviations used are:

AC - acyl group,

Ac is acetyl group,

AcOH - R>
BZI - benzyl group,

BSA-bovine serum albumin,

DIC - 1,3-diisopropylcarbodiimide,

DMEM medium Needle (EagIe), modified Dulbecco (Dulbecco);

Et is ethyl group,

EDTA-ethylenediaminetetraacetic acid,

FCBS serum bovine fetus

FMOC-9-fertilityscore,

For - formyl group,

HITE - Wednesday RPMII6 4D with the addition of 10-8M hydrocortisone, 5 μl/ml bovine insulin, 10 μg/ml human transferrin, 10-8M-estradiol in 310-8M Na2SeO3,

HOBt- 1-hydroxybenzotriazole,

HPLC - high performance liquid chromatography,

Me is a methyl group,

MeCN is acetonitrile,

MeOH methyl alcohol,

TEA - triethylamine,

PBS - phosphate-saline buffer solution,

PGlu - pyroglutamyl acid,

psi - pseudopeptide communication structure CH2-NH, except when the subsequent residue contains secondary N - terminal group, this then implies the education of the form CH2N,

TFA - triperoxonane acid,

Z - benzyloxycarbonyl.

In the present invention the most common polypeptides are polypeptides, the number and structure of which are mentioned below

1. NH<-psi-Leu-NH2< / BR>
4. 5F-D-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
5. D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
6. D-Tpi-Glu(OMa)-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
7. D-Tpi-His-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
8. D-Tpi-His(Bz)-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
9. NH2CO-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Phe-NH2< / BR>
10. D-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Phe-NH2< / BR>
11. D-Trp-Glu(MeNH)-Trp-Ala-Val-Gly-His-Leu-psi-Phe-NH2< / BR>
12. D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Phe-NH2< / BR>
13. D-Tpi-Glu(OMe)-Trp-Ala-Val-Gly-His-Leu-psi-Phe-NH2< / BR>
14. Hca-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
15. D-pGlu-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
16. Phe-Glu-Trp-Ala-Val-Gly-His-Leu-psi-Trp-NH2< / BR>
17. D-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
18. D-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
19. D-Trp-His(Bz)-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
20. D-Trp-Glu(MeNH)-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
21. D-Trp-Glu(OMe)-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
22. Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
23. Ac-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
24. NH2CO-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
25. Hna-Tpi-Gln-Trp-Ala-Val-Gly-Hls-Leu-psi-Tpi-NH2< / BR>
26. D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
27. Mpp-Gln-Trp-Ala-Gly-His-Leu-psi-Trp-NH2< / BR>
28. D-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp-NH2< / BR>
29. D-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp-NH2< / BR>
30. D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp-NH2< / BR>
31. Mpp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp(For)-NH2< / BR>
32. D-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp(For)rp-Ala-Val-Gly-His-Leu-psi-Tpi-OMe

36. D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-OMe

37. NH2CO-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-OMe

38. D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NHMe

39. D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-OH

40. D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi - N2H2CONH2< / BR>
In the present invention is particularly preferred polypeptides are peptides with numbers 5, 7, 12, 17, 18, 22, 26 and 38.

Synthesis of polypeptides

Polypeptides that meets the present invention can be obtained by any means known to the specialists working in the field of peptide chemistry. An overview of known methods can be found in the book M Bodanzky (M. Bodanszky Principle of Peptide Synthesis, Springer-Verlag, Heidellerg, 1984).

Methods specific to solid-phase synthesis, are described in the Handbook of Stewart and young (Y. M. Stewart and J. D. Young, Solid Phase Peptide Synthesis, Pierce Chem. Co., Rockford, IL, 1984 (2 nd ed)) and the baranyi and others (G. Barany, et al, Int. J. Peptide Protein Res., 30, 705-739, 1987).

A particularly preferred method of producing polypeptides and their intermediate peptides that meets the present invention is a method using solid-phase synthesis. The substrate used to carry out solid-phase synthesis of polypeptides that meets the present invention is benzhydrylamine resin or Florante protective group, selected for protection of the amino group, used tert-butoxycarbonyl group, which was removed at each stage of the synthesis. Source material containing a protected amino acid, was prepared from tert-butoxycarbonyl amino acids, coupled with benzylhydroxylamine resin or attached to chloroethylamino polystyrene resin by KF. The synthesis was started with the C-terminal moiety of the peptide and was performed using hardware manual steps, step by unlocking the alpha-amino group and the accession of the next amino acid.

Purification of polypeptides

Polypeptides typically purified by high pressure liquid chromatography on a column of reversed phase that was done on high-performance liquid chromatographic system of the firm "Rainin" (firm "Rainin Inc., Company", Woburn, PCs Minnesota), consisting of three pumps Rabbit HP HPLC firm "Rainin", computer-controlled firms "ARRL Macintosh place, injector firm "Readin and UV control device with variable wavelength model 87, manufactured by the firm "Noir". The crude peptides (10 - 40 mg) was loaded into a column of type Macro firm "Dynamics" (21.2 per 250 mm) at the and of the firm "Rainin Inc., Company") and suirable in the conditions of existence of a linear gradient, using a solvent system consisting of (A) 0.1% of triperoxonane acid and (B) 0,1% triperoxonane acid in 70% aqueous acetonitrile, at a speed of 2.0 ml/min In all fractions was estimated purity and retention time, for which we used are described below analytical high-performance liquid chromatographic system.

The quality and characteristics of elution of the crude and purified peptides was determined by analytical high-performance liquid chromatography, for which we used liquid chromatograph model 1090 manufactured by Hewlett-Canard" with dual beam detector, tuned to the wavelength of 220 and 280 nm, column reversed-phase (4,6 250 nm) containing gasket material W - porex C18(with a pore size of 300 and a particle size of 5 μm). In solvent systems A and B described above, the flow rate supported by the value of 1.2 ml/min, and the separation was carried out at room temperature.

In most cases, the additional polypeptides were purified, conducting re-chromatography using the same column, but with a slightly modified USLOVIE the homogeneity of the purified peptides exceeds 97%.

Amino acid analysis.

In the present invention the amino acid analysis of peptides was performed on the amino acid analyzer type 6300 manufactured by "Beckman", making it on the samples subjected to hydrolysis at 110oC for 20 h, which were in sealed evacuated tubes, which were introduced 4 M methansulfonate acid containing 0.2% 3-(2-amino-ethyl)-indole. The ratio of amino acids replied expected. Residues in the form of Leu-psi-Leu and Leu-psi-Phe gave absorption peaks with retention times, respectively 39,93 and 44,56 minutes In the adopted method of analysis education Tpi was not found after 50 min cleavage.

Analysis techniques.

A. Test for receptor binding

Binding of the drug125I-GRP (14-27) (gastronomically peptide with the number of members in the chain 14 - 27 and the label in the form of iodine-125) and displacement under the influence of anteromesial was carried out on plates for tissue culture with 24 grooves (GIBCO), using cell lines Swiss T. Fibroblasts lines Murine Swiss T were subjected to weekly environment DMEM containing 10% FCBS (serum bovine fetus) and antifungal drugs. Cultures were incubated at 37oC in air containing 5% Co2125I-GRP (14 - 27) in the absence or in the presence of different concentrations of antagonists (at concentrations 610-11-610-6M, the total volume of 0.4 ml).

According to Zachary and Rozengurt (Zachary and Rosengurd, Pres, Natl. Acad. Sci. USA, 1985, 85, 3636-3670) and Layton and others (Layton et al., Cancer Ris., 1988, 43, 4783 - 4789) binding of the drug125I-GRP at 37oC reaches the maximum value for 30 minutes and then subsides; thus, the cells were incubated at 37oC within 30 min After the cells were twice washed in chilled on ice (4oC) binding buffer and twice washed with chilled on ice phosphatebuffered saline (PBS, mm) of the following composition: NaCl 138, KCl - 2,8, Na2HPO4- 8, KH2PO4- 1,45, CaCl2- 0,91, MgCl2- 0,49. The washed culture was extracted with 0.5 M NaOH solution, taken in an amount of 0.5 ml, and transferred into vials DL is balali in the appropriate tube. Then the radioactivity of the samples was determined on an automatic gamma counter (firm Micromedic systems, Inc., , Huntsville, PCs Alabama).

To determine the types of receptor binding, the dissociation constant Kd, the Association constants Ka, the maximum binding capacity of the receptor Bmax and values premaxillae inhibition IC50used a computerized program coordination curves Ligand-PC proposed by Munson and Rodbard (Munson and Rodbard, Anal. Biochem. 1987, 107, 220 - 239).

The value of the IC50characterize concentrations of antagonists that cause premaxillae inhibition at GRP (14 - 27)-stimulated growth (at a concentration of 0.2 nm). The dissociation constant and maximum binding capacity of the125I-CRP (14 - 27) that occurred in the conducted experiments was to 1.32 nm 0,769 PM to 1 mg of protein, respectively, which is in agreement with similar values found in the literature for125I-GRP and125I-Tyr4-bombezin. Characteristics of the binding of GRP receptorow on the cell line T, obtained during the experiments are in good agreement with the values found for the binding of bombezin with pancreatic almosnino cells Jensen and others (J. iol. Chem., 1983, 258, 7527 - 7535).

Peptide GRP (14 - 27) inhibits binding 125I-GRP(14 - 27) with the value of the IC50equal 2,32 nm, which is in agreement with the data of Layton and others (Layton, et al, Cancer, Res., 1988, 48, 4783 - 4789), which lead value of 2.2 nm. Data binding polypeptides that meets the present invention shown in the attached table. 1.

The value of the IC50represents the concentration of unlabeled ligand that is half shifts the binding of specific radioligand. This value is calculated by the equation given by Cheng and ProSoft (Chend and Prusoff, Biochem. Pharmacol., 1973, 22, 3099), which has the following form:

IC50= Kc (1 + L/Kh), where Kc and Kh - dissociation constants, respectively ligand unlabeled (cold) and labeled (hot) and L is the concentration used radioligand.

B. Release of amylase

Selected pancreatic acini were prepared collagenase splitting of the pancreas taken from male rats Wistar (Wistar) (150-180 g), which were kept over night in the docked state. Animals were killed by cervical shift, and the pancreas was removed and then subjected to cleavage by means of purified collagenase and Amsterdam, Solomon and Jamieson (Amsterdam, Solomon and Jamieson, 1978).

Dispersed asinius suspended in incubation medium containing 24,5 HERES mm, 98 mm NaCl, 4.0 mm KCl, 11.7 mm KH2PO4, 1.0 mm MgCl2, 0.3 mm CaCl2, 5.0 mm glucose, 1% (weight per volume) mixture of essential and non-essential amino acids (firm "Serpa of Finmekkanika", , Heidelberg, Germany), 2 mm glutamine, and 0.2% bovine serum albumin (BSA) and 0.01% (weight per volume) trypsinogen inhibitor. The incubation solution was saturated with oxygen and kept at 37oC in the bath with shaking (60 oscillations per minute). Oceniony suspension were incubated in the presence of different concentrations gastronomiebedarf peptide (GRP) or antagonists gastronomiebedarf peptide.

After incubation the tubes were centrifuged at 1000 g for 5 min, and the precipitate was separated supernatant. The amylase content in the supernatant and dissolved in the sediment was determined separately that was done as described in Bernfeld (Bernfeld, 1955). The secretion of amylase was expressed as percentage of increment over the reference value. Incubation was repeated. Estimulando secretion of amylase during the whole experimental period took Zentrale was concentration dependent inhibition of the allocation process amylase, stimulated by submaximal concentrations gastronomiebedarf peptide (10-9M).

C. Inhibition capture3H-thymidine cell line T

Small cell lung cancer cells were used within 2-4 days after reseeding. Suspension of monoclonal cultures were prepared by washing cells twice washed in phosphate-saline buffer solution, and then the pipette transferred them in phosphate-saline buffer solution containing 0.2 g/l glucose, 0.2 g/l ethylendiaminetetraacetic acid and 14 mm chloride lignocaine) at 37oC what he was doing until, until the suspension became homogeneous (2-4 min). Cells are washed three times and resuspendable environment HITES without adding FCSB. Culture at day 0 were sown on the plate in the amount of 1,34105cells, then added all the peptides in 1 ml of RPMI-1640 medium containing environment HITES and 0.125% albumin. After 48 h in each recess was added 1 µci titiraupenga thymidine, and the incubation continued for another 24 h the Cells were then washed, put on filter paper of glass fiber and washed with chilled on ice with 50% trichloroacetic acid. Filter paper was placed in vials containing scinti gastronomiebedarf peptide (GRP) on the cell line T, inhibition capture thymine are presented in table.2.

D. Inhibition of growth of various small-cell lung cancer cells

Small cell lung cancer cells of strains H-69 and H-345, obtained from the National cancer Institute were transferred into the suspended state. The inhibition induced gastronomiebedarf peptide DNA synthesis occurring under the influence of anteromesial conducted by measuring the capture3H-thymidine. It was shown that the inhibition induced gastronomiebedarf peptide DNA synthesis occurring under the influence of anteromesial, is significant and is concentration dependent manner.

E. Effect on pancreatic secretion in vivo

Secretion studies were performed on six in consciousness cats (2-3 kg), prepared with the introduction of a permanent gastric and pancreatic fistulas that were done as described previously (Konturek et al., J. Discrimination London, 1976, 257, 663-672). In short, the cannula used in the gastric fistula, was of the type described previously in the literature (Emas, Gastroenterology, 1960, 39, 886-782). This cannula was introduced into the region of location of the pyloric gland near b the Noi form with a cross and main elements adapted by the inventors for cats. The common bile duct was dissected immediately in connection with the pancreatic ducts and transplanted to the upper part of the duodenum than was provided by the Department of bile flow from the flow of pancreatic juice. Was formed a small duodenal pocket containing the entrance of the main pancreatic duct, and the transverse element pancreatic cannula was introduced into this pocket. The main element of the cannula was placed in disanalogy duodenum approximately 3 cm below the duodeno-duodenostomy.

Secretory studies were started at the end of three months after surgery. Food was removed from cages at least 18 h prior to each experience. For each experience (excluding experience with breastfeeding) gastric fistula was left open than was provided escaping to the outside of the gastric juice.

The secret coming of pancreatic fistula were collected continuously and divided into samples with a dwell time of 15 minutes was Recorded volume, and determine the protein concentration and the bicarbonate and outputs that were done as described previously (Konturek et al., 1976).

Each animal is Svobody peptide was injected intravenously in increasing doses (1250 pmol/(AU) gastronomiebedarf peptide) in a one-day test with or without added peptide 5. In experiments with feeding gastric fistula was left closed, and each cat was given about 50 g kulinarne processed and homogenized by grinding meat, which is usually completely eaten up. Intravenous infusion of saline solution (about 10 ml/h) were made throughout the period after a meal, and when the pancreatic secretory response was found well defined plateau, was injected peptide 5, and secretion was studied during 2 hours In separate experiments hungry cats (without the introduction of peptide and feeding meat) main pancreatic secretion (with open gastric fistula) was measured within 2 h, and then the peptide 5 (10 nmol/(kg h)) was added in led environment with dose, which completely stopped pancreatic secretion, induced gastronomiebedarf peptide. These results are shown below.

Bombazine analogues in the form of peptides 5, 10, and 2 were subjected to testing in vivo for inhibition of serum gastrin after stimulation gastronomiebedarf peptide. After 8 min after stimulation gastronomiebedarf peptide (3 mg per 100 g of body weight) levels of serum gastrin increased with 16.7 PG/ml (control experiment) to 105 entered peptide antagonists 5, 10 and 2, was found to decrease the level gastrinomas secretion (injected 30 µg per 100 g of body weight; after 8 min was observed level of 36.8 PG/ml in the case of peptide 2, and 24.2 PG/ml in the case of peptide 10 and 39.2 PG/ml in the case of peptide 5).

The bombezin/GRP antagonists that meets the present invention are useful in the treatment of States of hypergastrinemia, for example, pernicious anemia, chronic atrophic gastritis syndrome Zollinger - Ellison and vitiligo, associated with diffuse hyperplasia of gastric enterochromaffin cells and an increased risk of developing primary multiple gastric carcinoids tumors. In addition, enterochromaffin cell hyperplasia easily produced in animals suffering from hypergastrinemia.

Such therapy has the advantage in comparison with existing drugs, since H2antagonists tend to cimetidine, which causes hypergastrinemia and can lead to the appearance of carcinoid tumors in humans. In addition, therapy is H2antagonists may lead to a relapse of ulcers in force the existing hypergastrinemia.

Since these compounds that meet this isiaka intestines and stomach cancer.

Based on these results above, data obtained in rats, peptides, corresponding to the present invention, can be introduced in the form of pharmaceutically acceptable nontoxic salts, such as salts obtained by addition of acids, are the chlorides, bromides, sulfates, phosphates, fumarate, gluconate, tannate, maleate, citrates, acetates, benzoate, succinate, alginates, pamoate, malaty, ascorbate, tartratami and similar substances.

Microcapsules and microparticles of these peptides formed from poly-DL - lactide-coglycolide, can be regarded as the preferred and carried out the forms for injection. Can also be applied intravenous, intramuscular or subcutaneous administration using isotonic saline, phosphate-saline buffer and similar solutions. For introduction into the lungs can also be used aerosols.

These pharmaceutical compositions should contain the peptide in conjunction with a conventional pharmaceutically acceptable carrier. Dosage should be about 1 to 1000 μg of peptide per 1 kg of body weight of the patient, if we are talking about parenteral way. Therapy of patients with these peptides can be carried out exactly as is to the s or other peptides.

These peptides can be introduced mammals intravenously, subcutaneously, intramuscularly, nasal or through the lungs in the form of an aerosol achievement gastric inhibition or anti-tumor effect. An effective amount of the dosage varies depending on the form of administration and the nature of the places that are processed at macpicasso. For example, a typical dosage form is a physiological saline solution containing the peptide, and this solution is injected in an amount such that the dose was in the range of from about 0.01 to 0.20 mg per 1 kg of body weight. The recipe can be discharged only once a month, and the duration of treatment can last several months.

Pharmaceutical composition for intramuscular injection long acting (sesame oil, gel)

Peptide 17 - 10.0 mg

The aluminum monostearate, USP - 20.0 mg

Sesame oil, g.s. to 1.0 ml

The aluminum monostearate is combined with sesame oil and heated to 125oC under stirring until then, until it forms a transparent yellow solution. This mixture is then sterilized in the autoclave and allowed to cool. Then in sterelny conditions by grinding into powder type peptide 17. Especially Privat activity for a long time.

Aqueous solution for intramuscular injection

Peptide 22 500 mg

Neoantigenic gelatin 5 mg

Water for injection, g.s. up to 100 ml

Gelatin and peptide 22 is dissolved in water for injection, then the solution is sterile filtered.

Microcapsules with prolonged activity for BM injection of the biodegradable polymer

Microcapsules prepared from the following ingredients:

Glycolic/lacheny (25/75) copolymer (characteristic viscosity of 0.5) - 99

Peptide 26 - 1

The above-mentioned microcapsules (25 mg) suspended in 1.0 ml of the following media, %:

Dextrose - 5

Sodium CMC (carboxymethylcellulose) - 0,5

Benzyl alcohol - 0,9

Twin 80 - 0,1

Water purified - 100

Although the invention was described with reference to its preferred options for implementation, it should be understood that changes and modifications, obvious to those having ordinary skill in the art may be made without departing for the scope of the invention described in the claims below. In the invention Pets possibility of replacement, known in this technical field that does not greatly affect its effectiveness.

General Operaciya-resin

Operation I:

1) rinse CH2Cl2(3 times for 1 min),

2) release 50% triperoxonane acid in CH2Cl2twice for 5 min and 25 min, respectively, in the case of peptide resins containing D - or L-Trp or Tpi, releasing spend 50% solution triperoxonane acid in CH2Cl2containing 5% mercaptoethanol and 5% anisole,

3) rinse CH2Cl2(6 times for 1 min),

4) neutralization of 10% solution of triethylamine in CH2Cl2(2 times for 3 min),

5) rinse CH2Cl2(6 times for 1 min),

6) articulation: (I) addition of BOC (butoxycarbonyl)-amino acid (3 equivalents) and HOBt (1-hydroxybenzotriazole) (3.3 equivalents) in dimethylformamide (3 minutes); (II) adding a 20% aqueous solution of diisopropylcarbodiimide (3 equivalent) in CH2Cl2and shake for 60 - 90 minutes,

7) rinse CH2Cl2(2 times for 1 min), ethylene (2 times for 1 min) and CH2Cl2(5 times for 1 min).

Operation II:

with the introduction of the recovered peptide bond-CH2NH-stage 6 I modify as follows:

1) washing with dimethylformamide (2 times for 1 min),

2) adding tert-butoxycarbonylamino aldehyde (3 equisetiformis and shake for 60 minutes,

4) rinsing with 50% methyl alcohol (3 times for 1 min), 100% methyl alcohol (3 times for 1 min) and CH2Cl2(3 times for 1 min).

Operations III.

Coupling of Boc-Asn, Boc-Gln and Boc-Gly stage 6 I modify as follows.

A solution of 20% diisopropylcarbodiimide (3 equivalents) are added to a mixture of dimethylformamide with tert-butoxycarbonylamino (3.0 equivalents) and 1-hydroxybenzotriazol (3.3 equivalents) at 0oC for 15 min and at room temperature for 15 insoluble substances are removed by filtration, the filtrate is added to the peptide resin and shaken with Boc-Gln or Boc-Asn within 2 to 4 h or Boc-Gly within 1 h

Operation IV

With the introduction of Fmoc(9-fertilityscore)-amino carry out the following stages:

1) after release and neutralize washed with CH2Cl2(3 times for 1 min) and dimethylformamide (3 times for 1 min),

2) the articulation of (I) the addition of Fmoc-amino acid (3 equivalents) and 1-hydroxybenzotriazole (3.3 equivalents) in dimethylformamide (3 min),

II) adding a 20% aqueous solution of diisopropylcarbodiimide (3 equivalent) in CH2Cl2and shaking for 60 min,

3) rinsing with ethanol (3 times for 1 min), di is of 30 minutes,

5) washing with dimethylformamide (6 times for 1 min),

6) another articulation is carried out as described in stage 2.

After getting the required intermediate peptides of formula I, the peptide resin is then treated with liquid HF in the presence of anisole, resulting in a gain polypeptide in a free form, which in the formula 1:X is hydrogen and Y is - NH2or OH, or a protected form, which has the formula I : A2- Glu(OMe) or His(Bz).

Treatment of functional groups in the N,C-terminal positions, or side chain groups of the polypeptide of free or protected form another N - or C-terminal group or side group of the functional group of the polypeptide carried out in solution, using an appropriate reagent. For example, blocking polypeptide containing the Glu group at

B. Example for L - and D-Boc-Tpi

To a stirred suspension consisting of 10.8 g (50 mm) D-Tpi 250 ml of 0.2 n NaOH solution and 7.5 ml of triethylamine, was added 10 g of di-tert-butylboronic, and the mixture was stirred for 4 h, then was added 10 g of bicarbonate, and after stirring for 3 h was added 10 more, the Mixture was stirred overnight and extracted (2 times 100 ml) simple ether, which was poured. Water is amywali water and dried in the air during the night.

Solids suspended in 100 ml of tetrahydrofuran. Dissolve almost all solids. Insoluble substances were removed by filtration and the tetrahydrofuran was removed under vacuum. The residue is triturated with simple ether, resulting in the received 9,20 g or 58% of the substance. This substance had the same melting point as the original substance, but differed in solubility and its characteristics when conducting thin-layer chromatography on silica using as eluent a mixture consisting of trichloromethane, methyl alcohol and acetic acid, taken in the ratio of 85 : 15 : 0.5 in.

Using the same technique from 2:55 g L-Tpi received 2,22 g or 59% Boc-Tpi.

2. Example obtain Boc-Leu-CHO

Methyl ester tert-butoxycarbonylamino (35 g, 134 mm) in dry toluene (250 ml) in a nitrogen atmosphere, was cooled with a mixture of dry ice with acetone, and for 30 min in toluene was added 25% Diisobutyl aluminum hydride (150 ml). The mixture was stirred for 20 min, which was done in a bath of dry ice with acetone after adding Diisobutyl aluminum hydride, then carefully added methanol (15 ml). The mixture was poured into ice water, taken in quantities of 1000 ml, the Aulnay and the ether extracts were combined and dried (sodium sulfate). The obtained oily liquid quickly passed through the column with a packing of silica gel (3 x 50 cm), for which we used 1500 ml of 15% mixture of ethyl ester of acetic acid in petroleum ether. Aldehyde Boc-Leu was received in the form of an oily liquid (27,6 g).

Example 2.

The number of the peptide and the structure below.

1. NH2CO-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
2. D-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
3. D-Trp-Gln-(MENH)-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
4. 5F-D-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
5. D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
6. D-Tpi-Gln(OMe)-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
7. D-Tpi-His-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
8. D-Tpi-His(Bz)-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
The polypeptides of this example containing fragments Trp-Ala-Val-Gly-His-Leu-Psi-Leu-NH2but having two different residue in N-terminal position, was built in stages on benzhydrylamine resin in accordance with standard methods for solid-phase synthesis.

Benzylhydroxylamine resin, taken in an amount of 0.50 g (0.9 mmole NH21 g), twice in three minutes was treated with 10% triethylamine in CH2Cl2(neutralization) and washed six times CH2Cl2. The resin was mixed with 1.35-mmol Boc-Leu and 1.50 isopropylcarbodiimide together with 1.3 mmol CH2Cl2. The mixture was shaken at room temperature for 60 minutes Obtained Boc-Leu-BHA (benzylideneamino) the resin was washed CH2Cl2what did twice with methanol, three times washed CH2Cl2and then subjected to the test according to Keyser (Kaiser, Anal Biochem., 34, 595 (1970)). In the case of incomplete coupling method for coupling was repeated.

Removal of Boc-group (release) of Boc-Leu-BHA-resin was carried out in solution in the form of a 50% triperoxonane acid in dichloromethane, which was done for 5 min, filtered, the processing is again repeated for 25 min and then washed six times with dichloromethane.

Neutralization of conducts as described above in the case of obtaining BHA-resin (benzhydrylamine resin).

The coupling of Boc-Leu=CHO was carried out by conducting the following operations II:

1) double washing with dimethylformamide,

2) add 1.5 mmole of Boc-Leu-CHO in dimethylformamide containing 1% acetic acid.

3) add 2.0 mmol NaBH3CN in dimethylformamide and shaken for 60 minutes,

4) rinsing with 50% methanol in water, twice with 100% methanol twice and CH2Cl23 times.

After removal of the Boc group from Boc-Leu-psi-Leu-BHA-resin and stoichiometric operation III.

A solution of 1,3-diisopropylcarbodiimide with a concentration of 20% (1.5 mmole) in CH2Cl2was added to a solution of dimethylformamide, consisting of 1.5 mmole of Boc-Gly and 1.65 mmole butanol and located at 0oC, was stirred with cooling for 15 min and at room temperature for 15 min, the precipitate was filtered, added to the resin and shaken for 60 minutes

Subsequent amino acid residues Boc-Val, Boc-Ala and Boc-Trp then successively introduced, cochleae by the method specified in the operations I, resulting in received from 0.90 g of the protected peptide resin with the structure of Boc-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Leu-BHA-resin (2/1 Res).

After enabling Ros-Trp were released Boc-group, which was affected by the 50% solution triperoxonane acid in dichloromethane containing 5% mercaptoethanol and 5% anisole, resulting received TFA Trp-Ala-Val-Glu-His(Z)-Leu-psi-Leu-BHA-resin (2/2 Res).

Education TFATrp-Ala-Val-Gly-His(Z)-Leu-psi-Leu-BHA-resin (2/2 Res), taken in the quantity of 0.91 g, were divided into eight portions (approximately 100 mg each), which were used to conduct synthesis design protected polypeptide resin in accordance with the techniques described in the operation I in relation to the coupling Boc-D-Trp, Boc-5F-D-Trp, Boc-D-Tpi and Boc-His(Z)Oh above heptapeptide resin (2/2 res) gives:

2/2/01 Boc-Trp-Gln-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Leu-BHA - resin.

Sequential coupling of Boc-Gln and Boc-D-Trp with heptapeptide resin (2/2/res) gives:

2/2/02 Boc-D-Trp-Gln-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Leu - BHA-resin.

The coupling of Boc-Gln and Boc-5F-D-Trp with heptapeptide resin (2/2/res) leads to:

2/2/04 Boc-5F-D-Trp-Gln-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Leu - BHA-resin.

Sequential coupling of Boc-Gln and Boc-D-Tpi get:

2/2/05 Boc-D-Tpi-Gln-Ala-Val-Gly-His(Z)-Leu-psi - Leu-BHA-resin.

Sequential coupling of Boc-Glu (OMe) and Boc-D-Tpi get:

2/2/06 Boc-D-Tpi-Glu(OMe)-Trp-Ala-Val-Gly-His(Z)-Leu - psi-Leu-BHA-resin.

Sequential coupling of Boc-His (Z)- and Boc-D-Tpi get:

2/07-Boc-D-Tpi-His(Z)-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Leu-BHA - resin.

Sequential coupling of Boc-His (Bz) and Boc-D-Tpi get:

2/2/08 Boc-D-Tpi-His(Bz)-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Leu-BHA - resin.

After removal of the Boc-group exposure is 50% triperoxonane acid in dichloromethane containing 5% mercaptoethanol and 5% anisole, Boc-released polypeptide resin was washed with dichloromethane, methanol and dichloromethane, which in each case do three times, and treated with svezheproseyannuyu HF (5 ml) and anisole (0.25 ml), what you are doing at 0oC for 1 h, the Solvent is distilled off under vacuum, washed with simple ether or ethylacetate product:

2/3/01 Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2.

The number of the peptide and the structure below.

4. D-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
5. 5F-D-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
6. D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
7. D-Tpi-Glu(OMe)-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
8. D-Tpi-His-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
2. D-Tpi-His(Bz)-Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2< / BR>
.

.

A mixture consisting of 40 mg of Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi - Leu-psi-Leu-NH2(2/3/1) and 20 μl of triethylamine, in 0.5 ml of dimethylformamide and 20 mg KOCN, in 100 μl of water, was stirred at 0oC, then the mixture was added dropwise 100 µl of acetic acid and stirred at 0oC for 1 h, the Reaction mixture was subjected to purification, resulting in a received peptide, the number and structure of which is given below:

1. NH2CO-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Leu - NH2.

Peptide 3 was prepared by sequential coupling with Fmoc-Gly(OBut) and Fmoc-D-Trp by the method specified in the operations IV, with Trp-Ala-Val-His(Z)-Leu-psi-Leu-BHA-resin (2/2/Res), resulting in the obtained Fmoc-D-Trp-Glu(OBut)-Trp-Ala-Val-His(Z)-Leu-psi-Leu - BHA-resin (2/4/3). The peptide resin was treated with 10% triperoxonane acid in dichloromethane containing 5% 2-mercaptoethanol, which was done within 30 minutes, cut estimateu washing with dichloromethane methylamine (MeNH2) propulsively through the layer of Fmoc-D-Trp-Glu-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Leu-BHA-resin (2/5/Res) in 5 ml of dimethylformamide, which was done at 0oC for 5 min, was added 0.25 ml of 20% 1,3-diisopropylcarbodiimide in dichloromethane and have interaction at 0oC for 2 hours and Then the resin was washed with dichloromethane and the Fmoc-group was removed with piperidine.

Peptide 3 D-Trp-Gly(MeNH)-Trp-Ala-Val-Gly-His-Leu-psi-Leu - NH2(RC-3490) was obtained after treatment of HF.

Purification was performed by high performance liquid chromatography using a solvent system consisting of (A) 0.1% of triperoxonane acid and (B) 1% triperoxonane acid in 70% acetonitrile. It turned out that the purified peptides obtained by analytical high-performance liquid chromatography, the purity of greater than 97%. The retention times of the polypeptides of this example are given in table. 3.

The results of amino acid analyses of polypeptides corresponding to this example, expected. For example, amino acid relations of peptide 2 with the structure of D-Trp-Gln - Trp-Ala-Val-Gly-His-Leu-psi-Leu-NH2was 1,11: 2,09:0,90: 1,03:0,95:0,92 (Gln:Trp:Ala:Val:Gly:His). The residue Leu-psi-Leu-detected absorption peak with a retention time 39,95 minize.

9. NH2CO-Tpr-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Phe-NH2< / BR>
10. D-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Phe-NH2< / BR>
11. D-Trp-Glu(MeNH)-Trp-Ala-Val-Gly-His-Leu-psi-Phe-NH2< / BR>
12. D-Tpi-Glu-Trp-Ala-Val-Gly-His-Leu-psi-Phe-NH2< / BR>
13. D-Tpi-Glu(OMe)-Trp-Ala-Val-Gly-His-Leu-psi-Phe-NH2< / BR>
.

.

Polypeptides corresponding to this example, contain the same fragment Trp-Ala-Val-Cly-His-Leu-psi-Phe-NH2. BOC-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Phe-BHA-resin (3/1/res) cost step by step what you did on 0.5 g benzhydrylamine resin (BHA resin) (9.0 mmole NH21 g) in accordance with the regulations for solid-phase synthesis, as described in the case of example 2, except that at the first junction take the fragment Boc-Phe instead of fragment Boc-Leu.

Part of the peptide resin, containing in each case, approximately 150 mg of Boc-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Phe-BHA-resin (3/1/Res), Sociali with the other two residues that were made by the techniques described in steps 1 applied to the coupling of Boc-Trp-Boc-D-Trp, Boc-D-Tpi and Boc-Glu (OMe) and operations III in relation to the articulation of Boc-Gln, which received final polypeptide resin.

Sequential coupling of Boc-Gln and Boc-Trp with the above heptapeptide resin (3/1/res) received:

3/2/09 Boc-Trp-Gln-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Phe-BHA-resin.

Consequently the BR>
Coupling of Boc-Gln and Boc-D-Tpi with heptapeptide resin (3/1/res) received:

3/2/12 Boc-D-Tpi-Gln-Trp-Ala-Val-Gly-His (Z)-Leu-psi-Phe-BHA-resin.

Coupling of Boc-Glu (OMe) and Boc-D-Tri with heptapeptide resin (3/1/res) build:

3/2/10 Boc-D-Tpi-Glu(OMe)-Trp-Ala-Val-Gly-His (Z)-Leu-psi-Phe-BHA-resin.

After removal of the Boc-group exposure is 50% triperoxonane acid in dichloromethane containing 5% mercaptoethanol and 5% anisole, polypeptide resin was washed with dichloromethane, methanol and dichloromethane, making it, in each case three times, and treated fresh HF (5 ml) and anisole (0.25 ml), which was done at 0oC for 1 h, the Solvent is kept in vacuum, were extracted 70-80% acetic acid and subjected to lyophilization. As a result of this received the following peptides:

3/3/09 Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Phe-NH2.

The number of peptide and structure below

10. D-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Phe-NH2< / BR>
12. D-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Phe-NH2< / BR>
13. D-Trp-Glu(OMe)-Trp-Ala-Val-Gly-His-Leu-psi-Phe-NH2< / BR>
.

.

The peptide containing the group NH2CO in N-terminal position, was prepared by the following method.

A mixture consisting of 40 mg of the crude polypeptide (3/3/9) Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Phe-NH2and 20 ál of triethylamine then dropwise added 100 μl of acetic acid, and the reaction when the stirring was conducted at 0oC for 1 h, the Reaction mixture containing the desired peptide is 9 kinds of NH2CO-Trp-Gln-Trp-Ala-Val - Gly-His-Leu-psi-Phe-NH2was subjected to purification by high performance liquid chromatography.

Peptide II was prepared by sequential coupling of two Fmoc-amino acids that were done by the method specified in the operations IV the solid-phase synthesis.

Resin TFA. Trp-Ala-Val-Gly-His(Z)-Leu-psi-Phe-BHA (3/1/Res), taken in an amount of 150 mg neutralized 10% triethylamine, washed with CH2Cl2and of dimethylformamide and Sociali with Fmoc-Glu (OBut), resulting in the obtained Fmoc-Glu(OBut)-Trp-Ala-Val - Gly-His(Z)-Leu-psi-Phe-BHA-resin (3/5/11). Fmoc-D-Trp-Glu(OBut)-Trp-Ala - Val-Gly-His(Z)-Leu-psi-Phe-BHA-resin was obtained after release of 50% piperidine in conjunction with Fmoc-D-Trp. Boutelou group (But) was removed from Fmoc-protected peptide resin by processing 10% triperoxonane acid in dichloromethane containing 2% mercaptoethanol, which lasted 30 minutes of Methylamide MeNH2propulsively through the layer (200 mg) resin Fmoc-D-Trp-Glu-Trp-Ala - Val-Gly-His(Z)-Leu-psi-Phe-BHA (3/6/11) in 5 ml of dimethylformamide, which was done at 0oC for 5 min, was added 0.2 ml of 20% 1,3-diisopropylcarbodiimide located in dichloromethane, and the mixture is displaced handling HF and anisole peptide 11 D-Trp-Glu(MeNH)-Trp-Ala-Val-Gly-His-Leu-psi-Phe - NH2was subjected to purification by high performance liquid chromatography.

The retention times of peptides corresponding to this example are shown in table. 4.

The relations among amino acids found in amino acid analysis, was responsible expected. For example, the relationship of peptide 10 type D-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Phe-NH2was 1,15:0,96:0,95:1,01:0,94: 1,97 (Glu: Gly: Ala: Val: His:Trp), and turned the peak with retention time of 44,56 minutes of Relations peptide 13 was 1,04:0,98:1,02:1,00:1,03:0,94 ((Glu:Cly:Ala:Val:His:Trp), and the observed absorption peak for Leu-psi-Phe with a retention time of 44,56 minutes Tpi Fragment of peptide 13 was not found.

Example 4.

The number of the peptide and the other below.

14. Hca-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
15. D-P-Glu-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
16. Phe-Glu-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
17. D-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
18. D-Trp-Gle-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
19. D-Trp-His(Bz)-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
20. D-Trp-Glu(MeNH)-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
21. D-Trp-Glu(OMe)-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
22. Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
23. Ac-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
24. NH2CO-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi - Tpi-NH2< / BR>
25. Hna-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
-Leu-psi-Trp-BHA-resin with formaldehyde in accordance with the methods below.

Boc-Leu-psi-Trp-BHA-resin was obtained from 0,1 benzhydrylamine (BHA) resin (0.9 mmole BH21 g) serial coupling with Boc-Trp and Boc-Leu-CHO, which was done by the method specified in the operation I and operation II. Dimethylformamide, taken in an amount of 10 ml containing 1% acetic acid, was added to the above peligni the resin, and then brought into interaction with 1 ml of 10% formaldehyde that was done at room temperature for 60 min, and washed with dimethylformamide, methanol and dichloromethane.

All polypeptides corresponding to this example, contain common code fragment Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2. Boc-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Tpi-BHA-resin (4/1/Res) built in stages on Leu-psi-Tpi-BHA-resin sequential coupling with Boc-His (Z) (I), Boc-Gly (III), Boc-Val, Boc-Ala and Boc-Trp (I).

Part of the above intermediate peptide resin in quantities of 150 mg was subjected to two further joints, making it according to the methods described in the operation I for articulation Hca, D-pGlu, Boc-Glu(OMe), Boc-Glu(OBz), Boc-D-Phe, Boc-D-Trp, Boc-His (Bz), Boc-Tpi, Boc-D-Tpi, AC-Tpi and Hna-Tpi and operations III for coupling Boc-Gln, resulting in the received end of the peptide resin.

Sequential coupling of Boc-Gln and Hca c above heptadienal Boc-Gln and D-pGlu to heptapeptide resin (4/1/res) received:

4/2/15 D-pGlu-Gln-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Tpi-BHA-resin.

Sequential coupling of Boc-Glu(OBz) and Boc-Phe with the above-mentioned intermediate peptide resin (4/1/res) received

4/2/16 Boc-Phe-Glu(OBz)-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Tpi-BHA-resin.

Coupling of Boc-Gln and Boc-D-Phe with heptapeptide resin (4/1/res) received:

4/2/17 Boc-D-Phe-Gln-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Tpi-BHA-resin.

Coupling of Boc-Gln and Boc-D-Trp with heptapeptide resin (4/1/res) build:

4/2/18 Boc-D-Trp-Gln-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Tpi-BHA-resin.

Coupling of Boc-His(Br) and Boc-D-Trp with heptapeptide resin (4/1/res) cost:

4/2/19 Boc-D-Trp-His(Br)-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Tpi-BHA-resin.

Coupling of Boc-Gln(MeO) and Boc-Tpi with heptapeptide resin (4/1/res) build:

4/2/21 Boc-D-Trp-Glu(OMe)-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Tpi-BHA-resin.

Coupling of Boc-Gln and Boc-Tri with heptapeptide resin (4/1/res) build:

4/2/22 Boc-Tpi-Gln-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Tpi-BHA-resin.

Coupling of Boc-Gln and Ac-Tpi (heptapeptide resin (4/1/res) build:

4/2/23 Ac-Tpi-Gln-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Tpi-BHA-resin.

Coupling of Boc-Gln and Hna-Tpi with heptapeptide resin (4/1/rec) build:

4/2/25 Hna-Tpi-Gln-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Tpi-BHA-resin.

Coupling of Boc-Glu and Boc-D-Tpi c heptapeptide resin (4/1/res) build:

4/2/26 Boc-D-Tpi-Gln-Trp-Ala-Val-Gly-His(Z)-Leu-psi - Tpi-BHA-resin.

After removal of the Boc-Uchaly the following peptides:

The number of the peptide and the structure below.

14. Hca-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
15. D-pGlu-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
.

4/3/16 Phe-Glu-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
.

17. D-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
18. D-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
19. D-Trp-His(Bz)-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
21. D-Trp-Glu(OMe)-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-Nh2< / BR>
22. Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
23. Ac-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
25. Hna-Tpi-Gln-Trp-Ala-Val-Glu-His-Leu-psi-Tpi-NH2< / BR>
26. D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2< / BR>
The peptide is Phe-Glu-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2(16) (4/3/16), taken in an amount of 20 mg together with 5 mg diphenylphosphinite and 10 mg KHCO3located in 0.5 ml of dimethylformamide, stirred at 0oC for 24 h the Reaction mixture was subjected to purification by HPLC using solvent system B (40-70%), which was done for 60 min, resulting in a received peptide 16:

Phe-Glu-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2,

the number of which amounted to about 4.5 mg) was purified (frequency higher than 95%) by analytical high-performance liquid chromatography using a solvent system at concentrations of 25-65%, DDA 22 species Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NH2, 20 μl of triethylamine, in 0.5 ml of dimethylformamide and 20 mg KOCN, in 100 μl of water, was stirred at 0 oC. After a few minutes, the above mixture was added dropwise 100 µl of acetic acid and the reaction with stirring was conducted at 0oC for 1 h, the Reaction mixture containing the desired (oligo) peptide: peptide 24 NH2CO-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi - Tpi-NH2was subjected to purification by high performance liquid chromatography.

Resin Fmoc-D-Trp-Gln(OBut)-Trp-Ala-Val-Gly-His(Z)- Leu-psi-Tpi-BHA (4/2/Res) was prepared by sequential coupling with Fmoc-Glu(OBut) and Fmoc-D-Trp to Trp-Ala-Val-Gly-His(Z)- Leu-psi-Tpi-BHA-resin (4/1/Res) according to the method specified in the operations IV. After removal, But the group's exposure to 10% triperoxonane acid in dichloromethane containing 2% of 2-mercaptoethanol, which was done for 30 min, the peptide resin was subjected to interaction with methylamine and 1,3-diisopropylcarbodiimide that was done according to the methods described in example 3 with respect to the peptide resin (3/6/11), resulting in the obtained Fmoc-D-Trp-Glu (MeNH)-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Tpi - BHA-resin (4/3/Res). After removal of the Fmoc - group exposure to piperidine derivatives of the peptide resin was treated with HF (5 ml) and anisole (0.25 ml what he was doing at 0oC for 1 h, in ratinov, meet the present example are shown in table. 5

When the amino acid analysis of peptides corresponding to the present example were obtained with the expected compositions. For example, polypeptide D-Phe-Gln-Trp-Ala-Gly-His-Leu-psi-Tpi-NH2(17) had a relationship 1,04:0,99:0,96: 1,00: 0,94: 0,99: 1,06 (Gln:Gly:Ala:Val:Phe:His:Trp). When the amino acid analysis of the Tpi fragment was not detected in peptides 17, 24 and 26.

Example 5

The number of the peptide and the structure below.

27. Mpp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp-NH2< / BR>
28. D-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp-NH2< / BR>
29. D-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp-NH2< / BR>
30. D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp-NH2< / BR>
31. Mpp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp(For)-NH2< / BR>
32. D-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp(For)-NH2< / BR>
33. D-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp(For)-NH2< / BR>
34. D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp(For)-NH2< / BR>
Peptides corresponding to this example, contain common code fragment Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp-NH2or Gln-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Trp (For) -NH2. Resin Boc-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp(For) -BHA (5/1/Res) created on 1.0 g benzhydrylamine resin (BHA resin) (0.9 mmole NH21 g) consistent articulation with the operations of the solid-phase synthesis described in example 2, except that at the first joint instead of Boc-Leu took Boc-Tr is protected peptide resins, who finally got a joint with MPP, Boc-D-Phe, Boc-D-Trp or Boc-D-Tpi respectively, made according to the method described in the operation I.

5/2/27. Mpp-Gln-Trp-Ala-Val-Gly-His(2)-Leu-psi-Trp(For)-BHA resin

5/2/28. Boc-D-Phe-Gln-Trp-Ala-Val-Gly-His(Z)-Leu-psi-p(For)-BHA resin

5/2/29. Boc-D-Trp-Gln-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Trp(For)-BHA resin

5/2/30. Boc-D-Tpi-Gln-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Trp(For)-BHA resin

After removal of the Boc-group exposure is 50% triperoxonane acid in dichloromethane containing 5% mercaptoethanol and 5% anisole, half of each of the above peptide resin was treated with HF (5 ml) and anisole (0.25 ml), which was done at 0oC for 1 h, resulting received the following peptides:

The number of peptide and structure below

31. Mpp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp(For)-NH2< / BR>
32. D-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp(For)-NH2< / BR>
33. D-Trp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp(For)-NH2< / BR>
34. D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp(For)-NH2< / BR>
The remaining half of each peptide resin was treated with hydrofluoric acid containing 5% anisole and 5% dimercaptotoluene that was done at 0oC for 1 h, resulting received the following peptides:

The number of the peptide and the structure below.

27. Mpp-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Trp-NH2< / BR>
28. D-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-psi was purified by high performance liquid chromatography. The obtained retention times are given in table. 6.

Data obtained by amino acid analysis of peptides corresponding to this example, expected. For example, the peptide 28 had the following amino acid relations: 0,98:0,92:1,03:0,97:0,98:1,09 (Gly:Ala:Val:Phe:His:Trp). The Tpi fragment the peptides 30 and 34 was not found.

Example 2

An example of a peptide and the structure below.

35. Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-OMe

36. D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-OMe

37. NH2CO-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-OMe

38. D-Tpi-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-NHMe

39. D-Tpi-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-OH

40. D-Tpi-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-N2H2CONH2< / BR>
As source material used Boc-Trp - OCH2-resin which was prepared by the following method. A mixture consisting of ClCH2-resin (1.0 g 0.7 mmole of chlorine per 1 g) Boc-Trp (2.0 mmole Trp) and KF (4 mmole) in 20 ml of dimethylformamide: was stirred at 70 - 80oC for 4 h and Then Boc-Trp-OCH2the resin is washed twice using in each case the methanol, water, methanol, dimethylformamide and dichloromethane. Resin Boc-Leu-psi-Trp-OCH2was obtained by coupling of Boc-Leu-CHO with Trp-OCH2-resin, by operation II. Resin Boc-Leu-psi-Tpi-OCH2was obtained by interaction of Boc-Leu-psi-Trp - OCH2), Boc-Gly-Boc, Val-Boc-Ala-Boc-Trp and Boc-Gln conducted on the operations of the solid-phase synthesis, as described previously received 1,60 Boc-Gln-Trp-Ala-Val-Gly-His(Z)-Leu-psi-Tpi - OCH2-resin (6/1/Res). Part of the above intermediate peptide resin was used to obtain Boc-Tpi-Gln-Trp - Ala-Val-Gly-His(Z)-Leu-psi-Tpi-OCH2-resin (6/2/35), which was achieved by coupling with Boc-Tpi. Another aliquot of the peptide resin was used to obtain Boc-D-Tpi-Gln-Trp-Ala-Val-Gly-His(Z)- Leu-psi-Tpi-OCH2-resin (6/2/36), which was achieved by coupling with Boc-D-Tpi.

After removal of the Boc-group exposure is 50% triperoxonane acid found in diethylmaleate (DEM) containing 5% mercaptoethanol and 5% anisole, transesterification was carried out as follows. Mixed 0.5 g Tpi-Gln-Trp-Ala-Val-Gly-His(Z)-Leu - psi-Tpi-OCH2resin (6/3/35), methanol (15 ml) in dimethylformamide (15 ml) and diisopropylethylamine (3 ml) and the mixture was stirred at room temperature for three days. The resin was washed with dimethylformamide (3 times) and methanol (3 times). The filtrate and wash water were combined and evaporated on a rotary evaporator in a vacuum, that the items in order to remove solvents. After processing the HF and anisole were received 123 g crude peptide 35

Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-OCH3.

Peptide 36 D-Tpi-Gln-Trp-Ala-Val-Gly-His(Z)-Leu-psi - Tpi-OCHS="ptx2">

A mixture consisting of Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi - Tpi-OCH3(peptide 35) (40 ml), 20 μl of triethylamine, in 0.5 ml of dimethylformamide, and 50 mg of compound KOCN, in 100 μl of water, was stirred at 0oC, after a few minutes was added 50 μl of acetic acid, and the interaction was conducted at 0oC for 1 h the Mixture was then subjected to purification, resulting in a received peptide 37

NH2CO-Tpi-Gln-Trp-Ala-Val-His-Leu-psi-Tpi-OCH3< / BR>
A mixture consisting of D-Tpi-Gln-Trp-Ala-Val-Gly-His(Z)-Leu - psi-Tpi-OCH3(peptide 36) and a solution of methylamine in methanol (2 ml) taken in a weight ratio of 1: 2, was stirred under room temperature for 16 hours After evaporation on a rotary evaporator under vacuum the residue was dried by freezing and was treated with HF and anisole. Received the product in the form of peptide 38 D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu - psi-Tpi-NHCH3, which was subjected to purification by the method of high performance liquid chromatography.

Another part of the resin D-Tpi-Gln-Trp-Ala-Val-Gly-His(Z)-Leu - psi-Tpi-OCH2(6/2/35) were subjected to interaction with HF and anisole, resulting received

peptide 39 D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi-OH.

The mixture of peptide 39 (40 mg), (Boc)2O (20 mg) and triethylamine (0,5 μl) in 0.5 ml of dimethylformamide PE the IDT with 1-hydroxybenzotriazole (10 mg) and N2H3CONH2(20 mg) were subjected to interaction with DCl (100 μl, 20% solution of DCl in dichloromethane), which was done over night at 0oC, the dimethylformamide was evaporated, washed with simple ether and removed the Boc-group, affecting 50% triperoxonane acid containing 5% mercaptoethanol and anisole, resulting received crude peptide 40

D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leu-psi-Tpi - N2H2CONH2.

1. Derivatives nonapeptides General formula I

X - A1- A2- Trp- Ala - Val - Gly - His - Teu - Tpi psi Q,

where Q - Ome, NH2;

X is hydrogen, acetyl, CONH2;

A1(D - or L-)Phe, D-Trp, D - or L-) - pi, D-p-Glu;

A2- Gln, Gly (OMe), Glu (MeNH), Glu, His (BZ)

or their salts with pharmaceutically acceptable acids.

2. The nonapeptide under item 1 of the formula

D - Phe - Gln - Trp - Ala - Val - Gly - His - eu - Tpi psi NH2.

3. The nonapeptide under item 1 of the formula

D - Phe - Glu - Trp - Ala - Val - Gly - His - eu - Tpi psi NH2.

4. The nonapeptide under item 1 of the formula

D - Trp - Gln - Trp - Ala - Val - Gly - His - eu - Tpi psi NH2.

5. The nonapeptide under item 1 of the formula

Trp - Gln - Trp - Ala - Val - Gly - His - eu - Tpi psi NH2.

6. The nonapeptide under item 1 of the formula selected from the group

NH2CO Tpi - Gln - Trp - Ala - Val - Gly - His - eu - Tpi psi NH2.

CH32
.

8. The nonapeptide under item 1 of the formula

D - Trp - Glu(OMe) - Trp - Ala - Val - Gly - His - eu - Tpi psi NH2.

9. The nonapeptide under item 1 of the formula

D - Trp - Glu(NHMe) - Trp - Ala - Val - Gly - His - eu - Tpi psi NH2.

10. The nonapeptide under item 1 of the formula

D - Trp - His(BZ) - Trp - Ala - Val - Gly - His - eu - Tpi psi NH2.

11. The nonapeptide under item 1 of the formula I in which A is D-or L - Tpi.

12. Derivatives nonapeptides General formula I under item 1, which has antagonistic activity against bombezin.

13. Derivatives nonapeptides General formula II

D - Tpi - A2- Trp - Ala - Val - Gly - His - eu - psi - A 9 NH2,

where A2Is Gln, Glu (OMe), His, His (Bz);

A9eu, Phe, Trp, Trp (For),

or their salts with pharmaceutically acceptable acids.

14. Derivatives nonapeptides on p. 2 of the formula

D - Tpi - Gln - Trp - Ala - Val - Gly - His - eu - eu psi NH2.

15. Derivatives nonapeptides on p. 2 of the formula

D - Tpi - Gln - Trp - Ala - Val - Gly - His - Leu - psi Phe NH2.

16. Derivatives nonapeptides on p. 2 of the formula

D - Tpi - Gln - Trp - Ala - Val - Gly - His - Leu - psi TrpNH2.

17. Pharmaceutical composition having antagonistic activity against bombezin containing the active ingredient and pharmaceutically acceptable carrier, featuring the number.

18. The method of obtaining derivatives of nonapeptides General formula I under item 1, which consists in a stepwise extension of the peptide chain, which is carried out by protecting the amino group, in - position, and the end carboxypropyl covalently bind with a synthetic polymer carrier used for this purpose, after which protecting group otscheplaut, carboxypropyl subsequent amino acids to restore education group

which then bind with the amino group, subjected to the pre-release, then aminoamides group of the second amino acid will unlock and carboxypropyl subsequent amino acids are then associated with the released - amino group of a second amino acid, and then the step in the sequence corresponding to structural formula I, attach other amino acids, obtained from peptidyl-polymer otscheplaut media and, if necessary, the protective group of the obtained peptides if necessary acelerou and the target product is isolated in free form or in the form of a salt with a pharmaceutically acceptable acid.

19. The method of obtaining derivatives of nonapeptides General formula II under item 13, which consists in a stepwise increase of the peptide associated with the synthetic polymer carrier, used for this purpose, after which protecting group otscheplaut, carboxypropyl subsequent amino acids to restore education group

< / BR>
which then bind with the amino group subjected prior to release, then aminoamides group of the second amino acid will unlock and carboxypropyl subsequent amino acids are then associated with the released - amino group of a second amino acid, and then the step in the sequence corresponding to structural formula II, attach other amino acids, obtained from peptidyl-polymer otscheplaut resin carrier and, if necessary, the protective group and the target product is isolated in free form or in the form of a salt with a pharmaceutically acceptable acid.

 

Same patents:

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)

sleep" target="_blank">

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 the chemistry of peptides, particularly to a method of obtaining nonpartitioned with a strong LH-RH/FSH-RH activity

The invention relates to the chemistry of peptides, or rather to Decapeptide, analogue releasing factor, luteinizing and follicle-stimulating hormone - luliberin with antitumor activity in experimental models of hormone-dependent tumors

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

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 the chemistry of peptides, or rather to Decapeptide, analogue releasing factor, luteinizing and follicle-stimulating hormone - luliberin with antitumor activity in experimental models of hormone-dependent tumors

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

Up!