Peptide with immunomodulatory activity
(57) Abstract:The invention relates to medicine, namely to compounds with immunomodulatory properties. As immunomodulator proposed peptide patterns
< / BR>where R is H, lower acyl or lower alkyl, and X is an aromatic or heteroaromatic amino acid or its derivative. 3 table. The invention relates to medicine, namely to new peptide structures with immunomodulatory properties.Known medical use of a large number of Immunostimulants, such as drugs ginseng and schisandra, derivatives of nicotinic acid, thymalin and so on (Mashkovsky M. D. Medicines, T. 11, M, Medicine, 1988, S. 169).The disadvantage of most natural products is low activity due to the low content of active principle, the presence of such effects.We know a significant number of Immunostimulants peptide, for example, taktivin (French patent N b, class A 61 K 39/41), thymosin (Goldstein A. L. et al, Proc. Nat. Acad. Sci. USA, 1972, v. 69, p. 1856-1863), obtained from natural raw materials. These drugs constitute a set of polypeptides of different lengths, affecting nulinio unwanted side effects. In addition, the practical use of such drugs is difficult due to the complexity of the methods of their extraction from natural raw materials, low output, significant variability in their physical and chemical properties.Search synthetic peptides with selective activity on account of changes in the structure would be more appropriate. Synthetic peptide Immunostimulants are the peptide Glu-Asp-Ser-Ser-Thr-Gly-Trp-Asp-OH (US patent N 4478828, class A 61 K 37/00, C 07 C 103/52, 1984), analogues thymopentin (European patent class. C 07 K 7/64, 1985, US patent N 5013723, class A 61 K 37/02, 1991), etc.Even more interesting is the creation of a peptide of Immunostimulants on the basis of short selectively active highly active peptides, the synthesis of which is economically advantageous.The prototype of the claimed invention was the peptide H-L-Glu-L-Trp-OH, the lack of which is less high immunomodulating activity.The problem faced by the authors in creating the present invention was to develop more effective and safe immune modulators on the basis of short peptides.It was found that this problem can be solved by using at least one of the peptides aminosilica or its derivative, for example ester, amide, hydrazide, etc.Peptides receive standard technology, peptide synthesis on solid phase (J. M. Steward and J. D. Young, Solid Phase Peptide Synthesis, W. H. Freeman Co. San Francisco, 1969), or by synthesis in solution (E. Schroder and K. Lubke, The Peptides, Vol. 1, Academic Press (New York), 1965) followed by removal of protective groups and purification of the final product.As an interim protection-amino group of amino acids for solid-phase method of synthesis using tert-butyloxycarbonyl group. For protection of side radicals of amino acids using the following groups: for histidine benzyloxyethanol; tyrosine 2,6-dichlorobenzidine; formyl tryptophan, etc. For condensation of use dicyclohexylcarbodiimide or dicyclohexylcarbodiimide with the addition of 1-hydroxybenzotriazole. Release are 50% solution triperoxonane acid in methylene chloride and neutralizing with 5% solution of diisopropylethylamine in methylene chloride.Cleavage of these peptides from the resin and release the side of the protective groups are using anhydrous hydrogen fluoride, and the cleansing - gel - and obremenitve chromatography.Example 1. Synthesis of peptide g-L-glutamyl-L-tryptophan (1)
0,6 teleformula, add 0.2 g (0,0018 mol) N-oxysuccinimide, cooled to minus 5oC with vigorous stirring poured to the reaction mixture a solution of 0.37 g (0,0018 mol) of N,N-dicyclohexylcarbodiimide 2.0 cm3of dimethylformamide. The mixture was stirred at 0oC 1 h and left for 12 h at room temperature. Filtered dicyclohexylamine, added to the filtrate to 0.72 g (0,0022 mol) of the hydrochloride benzyl ester of L-tryptophan and 0.3 cm3(0,0023 mol) of triethylamine and the reaction mixture is stirred 16 h at room temperature. The solution is filtered, diluted with 50.0 cm3water and extracted three times with 40 cm3ethyl acetate. The combined organic extracts are washed sequentially 20.0 cm3water, twice 20.0 cm32 n sulfuric acid, twice 20.0 cm3a saturated solution of sodium chloride and dried with anhydrous sodium sulfate.Evaporated the organic solution under reduced pressure, the residue is treated 16,0 cm350%-aqueous solution triperoxonane acid in methylene chloride for 45 min, again evaporated under reduced pressure and added 25,0 cm3isopropyl alcohol.To the obtained solution of 0.3 g (0,00048 mol) of fluoroacetate demonia. The reaction mixture is heated to 50oC, is added in small portions 5.0 g of 10% palladium on coal, suspended in 25,0 cm3water, and stirred for 30 minutes, Filtered off the catalyst, evaporated isopropyl alcohol under reduced pressure, an aqueous solution lyophilized, was added to the residue 20.0 cm3water and again lyophilized.The obtained freeze-dried purified by reversed-phase chromatography on a column of Ultraprep C18, gradient (10-30%) of acetonitrile in 0.1% triperoxonane acid. The content of the basic substance according to the optical density not less than 97% Structure confirmed by amino acid analysis of hydrolyzed peptide obtained by hydrolysis of 3 N. paratoluenesulfonyl. The analysis revealed the following relationship amino acid residues: Glu 1,0(1); Trp 0,9(1).Example 2. Synthesis of peptide g-L-glutamyl-Nin-formyl-L-tryptophan (II)
For the synthesis of peptide g-L-glutamyl-Nin-formyl-L-tryptophan is loaded into the reaction vessel 0.6 g of tert-butyloxycarbonyl-Nin-formyl-L-tryptophyl-polymer. The content of tryptophan 0.50 mmol/g of polymer. Peptide chain further increase according to the following program (see tab.1).If the ninhydrin test is positive, pogo residue use a-benzyl ester N-carbobenzoxy-L-glutamic acid.Upon completion of the synthesis, the peptidyl-polymer dried in a vacuum desiccator over pjatiokisi phosphorus to constant weight. 1 g of peptidyl-polymer is transferred to the digester to work with liquid hydrogen fluoride, add 1 ml of thioanisole and 1 ml of meta-cresol, cooled to a temperature of -78oC and for 10 min distilled 10 ml of anhydrous hydrogen fluoride. The reaction mixture is heated to 0oC and stirred for 60 min, then distilled hydrogen fluoride. The residue is transferred to the filter SCHOTT and washed with ethyl acetate and ether. The peptide extracted with 50 ml of 25% acetic acid, diluted with water and lyophilizers.After desalting on a column with Sephadex G-10 obtained lyophilized purified by reversed-phase chromatography on a column of Ultraprep C18, gradient (10-30%) of acetonitrile in 0.1% triptocaine acid. The content of the basic substance, according to the optical density, at least 96% Amino acid composition of the peptide corresponds to theoretical.Example 3. Synthesis of peptide g-L-glutamyl-b-thienyl-D-alanyl-amide (III).Synthesis, blocking, purification and characterization of the peptide are carried out in the same way that the peptide of example 2, except that the reaction vessel sagrei the release to the environment of anhydrous hydrogen fluoride for 2 hours The content of b-thienyl-D-alanine 0.5 mmol/g of polymer.The content of the basic substance, according to the optical density not less than 97%
Example 4. Synthesis of peptide N-methyl-g-L-glutamyl-L-tryptophan (IV).Synthesis of peptide (IV) is carried out in the same way that the peptide (II) of example 2, except that at the stage of condensation as the joining of amino acid residue use a-benzyl ester N-tertbutyloxycarbonyl N-methyl-L-glutamic acid.Upon completion of the synthesis, the peptidyl-polymer dried in a vacuum desiccator over pjatiokisi phosphorus to constant weight. 1 g of peptidyl-polymer is transferred to the digester to work with liquid hydrogen fluoride, added to 6.5 ml of dimethyl sulfide, 0.8 ml of meta-cresol and 0.4 ml identicial, cooled to a temperature of -78oC for 10 min, 2.5 ml distilled anhydrous hydrogen fluoride. The reaction mixture is heated to 0oC and stirred for 120 min, then distilled hydrogen fluoride and dimethyl sulfide. The residue is transferred to the filter SCHOTT and washed with ethyl acetate and ether. The peptide extracted with 50 ml of 25% acetic acid, diluted with water and lyophilizers.After desalting on a column with Sephadex G-10 Rila 0.1% triptocaine acid. The content of the basic substance, according to the optical density not less than 97% of the Structure of matter was substantiated by the conditions of synthesis and amino acid composition. The amino acid composition of the peptide corresponds to theoretical.Example 5. Synthesis of peptide N-acetyl - g-L-glutamyl-L-tryptophan (V).Synthesis of peptide (IV) is carried out in the same way that the peptide (II) of example 2, except at the stage of condensation as the joining of amino acid residue use a-benzyl ester N-tertbutyloxycarbonyl L-glutamic acid. Dipeptidyl-polymer handle 50% triperoxonane acid in methylene chloride (2 times, 2 and 30 min), washed with methylene chloride (4 times, 2 min) and incubated in a mixture containing of 0.11 ml of triethylamine, and 0.28 ml of acetic anhydride and 5 ml of methylene chloride.Upon completion of the peptidyl-polymer was washed with methylene chloride (6 times for 2 min), dried in vacuum, will unlock and clear, as shown in example 4 (peptide IV). The content of the basic substance, according to the optical density, at least 96% Amino acid composition of the peptide corresponds to theoretical.Immunomodulating activity of the claimed peptides were evaluated in tests of their wlia the main growth factor of T-cell interleukin-2.Example 6. The effect of peptides on the differentiation of precursor T-cell bone marrow of mice.A suspension of bone marrow cells of mice succumbing dislocation sewing vertebrae, obtained by flushing femurs with saline. Peptides at various concentrations is added to the received cells in the environment of the Needle and incubated for one hour at 37oC. the Level of expression of a marker of differentiation of T-lymphocytes q-antigen define a method complementation cytolysis using antibodies to q-antigen (Terasabi P. I. et al. in Manual of Tissue Typing Techniques, Nathional Institute of Health, Bethesda, 1972, p. 50-55). The data are given in table.2.Example 7. The effect of peptides on the production of interleukin-2 by mouse splenocytes.Data on the influence of peptides on the induction of synthesis of the main growth factor T-cell interleukin-2 (Gillis, S. et. al. J. Immunol. 1978, v. 120, p. 2027-2032) are given in table. 3.Drugs are not toxic, as 1000-fold excess of therapeutic dose (1 mg/kg) death was not detected in any one animal. The peptide of General formula
< / BR>where R is H, lower acyl or lower alkyl;
and X is an aromatic or heteroaromatic amino acid or its proizvodi the
< / BR>the way they are received and to farbkomposition based on them
FIELD: organic chemistry, medicine.
SUBSTANCE: invention relates to applying compounds of the formula (I) for preparing an antibacterial composition and veterinary composition eliciting with the enhanced activity.
EFFECT: valuable properties of agents.
4 cl, 3 tbl, 78 ex
FIELD: organic chemistry, biochemistry, medicine, pharmacy.
SUBSTANCE: invention relates to macrocyclic peptides of the general formula (I): wherein W means nitrogen atom (N); R21 means hydrogen atom (H), (C1-C6)-alkoxy-, hydroxy-group or N-(C1-C6-alkyl)2; R22 means hydrogen atom (H), (C1-C6)-alkyl, CF3, (C1-C6)-alkoxy-group, (C2-C7)-alkoxyalkyl, C6-aryl or Het wherein het means five- or six-membered saturated or unsaturated heterocycle comprising two heteroatoms taken among nitrogen, oxygen or sulfur atom and wherein indicated Het is substituted with radical R24 wherein R23 means hydrogen atom (H), -NH-C(O)-R26, OR26, -NHC(O)-NH-R26, -NHC(O)-OR26 wherein R26 means hydrogen atom, (C1-C6)-alkyl; R3 means hydroxy-group or group of the formula -NH-R31 wherein R31 means -C(O)-R32, -C(O)-NHR32 or -C(O)-OR32 wherein R32 means (C1-C6)-alkyl or (C3-C6)-cycloalkyl; D means a saturated or unsaturated alkylene chain comprising of 5-10 carbon atoms and comprising optionally one-three heteroatoms taken independently of one another among oxygen (O), sulfur (S) atom, or N-R41 wherein R41 means hydrogen atom (H), -C(O)-R42 wherein R42 means (C1-C6)-alkyl, C6-aryl; R4 means hydrogen atom (H) or one-three substitutes at any carbon atom in chain D wherein substitutes are taken independently of one another from group comprising (C1-C6)-alkyl, hydroxyl; A means carboxylic acid or its alkyl esters or their derivatives. Invention relates to pharmaceutical compositions containing indicated compounds and eliciting activity with respect to hepatitis C virus and these peptides inhibit activity of NS3-protease specifically but don't elicit significant inhibitory activity with respect to other serine proteases.
EFFECT: valuable biochemical and medicinal properties of peptides.
106 cl, 9 tbl, 61 ex
FIELD: organic chemistry, medicine, pharmacy.
SUBSTANCE: invention relates to compounds of the formula (I):
wherein r = 1, 2 or 3; s = 0; t = 0; R1 is taken among group including R11-CO and R12-SO2- wherein R11 is taken among group including (C6-C14)-aryl, (C1-C8)-alkyloxy-group wherein all given group are unsubstituted or substituted with a single or some similar or different substitutes R40; R12 means (C6-C14)-aryl wherein indicated group is unsubstituted or substituted with a single or some similar or different substituted R40; R2 means R21(R22)CH-, R23-Het-(CH2)k-, R23(R24)N-(CH2)m-D-(CH2)n- or R25(R26)N-CO-(CH2)p-D-(CH2)q- wherein D means bivalent residue -C(R31)(R32)-, bivalent (C6-C14)-arylene residue or bivalent residue obtained from aromatic group Het comprising 5 or 6 atoms in cycle among them 1 or 2 are similar or different cyclic heteroatoms taken among group including nitrogen and sulfur atoms; numbers k, m, n, p and q = 0, 1, 2; R21 and R22 that are independent of one another can be similar or different and taken among group including hydrogen atom, (C1-C12)-alkyl, (C6-C14)-aryl and so on; R23 means hydrogen atom, R27-SO2- or R28-CO-; R24, R25 and R26 mean hydrogen atom; R27 is taken among group including (C1-C8)-alkyl, (C6-C14)-aryl and so on; R28 is taken among group including R27, (C1-C8)-alkyloxy-group; R31 and R32 mean hydrogen atom; R40 is taken among group including halogen atom, hydroxy-, (C1-C8)-alkyloxy-group, (C1-C8)-alkyl, (C6-C14)-aryl and so on; R91, R92, R93 and R96 means hydrogen atom; R95 means amidino-group; R97 means R99-(C1-C8)-alkyl; R99 is taken among group including hydroxycarbonyl- and (C1-C8)-alkyloxycarbonyl-; Het means saturated, partially unsaturated or aromatic monocyclic structure comprising from 3 to 6 atoms in cycle among them 1 or 2 are similar or different heteroatoms taken among group comprising nitrogen and sulfur atoms; in all its stereoisomeric forms and also their mixtures in any ratios, and its physiologically acceptable salts. Invention proposes a method for preparing compound of the formula (I). Also, invention proposes a pharmaceutical preparation eliciting inhibitory activity with respect to factor VIIA and containing at least one compound of the formula (I) and/or its physiologically acceptable salts and pharmaceutically acceptable carrier. Invention provides preparing compounds of the formula (I) eliciting power anti-thrombosis effect and useful for treatment and prophylaxis of thrombosis-embolic diseases.
EFFECT: valuable medicinal properties of compounds and composition.
10 cl, 70 ex
FIELD: organic chemistry and drugs.
SUBSTANCE: New class of compounds of general formula 1, where R has formula 2 or 3; other residues are as described in claim of invention is disclosed. Said compounds are interleikyn-1β converting enzyme (ICE) inhibitors and have specific structural and physicochemical properties. Invention also relates to pharmaceutical composition containing said compounds. Compounds and composition of present invention are particularly useful in ICE activity inhibition and thereby can be used as drug for treating of diseases mediated by IL-1, apoptosis, IGIF and IFN-γ, as well as inflammations, autoimmune diseases, bone-destructive disorder, infections, disorder associated with cell proliferation, degenerative and necrotic disorders. Uses of claimed compounds and compositions as well as methods for production of N-acylamino compounds also are disclosed.
EFFECT: effective interleikyn-1beta converting enzyme inhibitors.
64 cl, 35 ex, 35 tbl, 21 dwg
FIELD: medicine, gastroenterology.
SUBSTANCE: traditional eradication therapy should be supplemented with licopid at the dosage of 10 mg per os once daily before breakfast for 10 d. The present innovation prevents transfer of microorganisms into inactive form, accelerates restoration of mucosal epithelial layer in gastroduodenal area, provides complete eradication of microorganisms, that in its turn, favors to prevent disease exacerbation and restoration of gastroduodenal functions.
EFFECT: higher efficiency of therapy.
3 dwg, 2 ex
FIELD: biotechnology, biochemistry.
SUBSTANCE: invention relates to producing the biologically active complex eliciting antioxidant and immunomodulating activity and used in medicine, cosmetics, veterinary science and food industry. The biologically active complex preparing by enzymatic hydrolysis of muscle tissue represents complex of biologically active compounds involving carnosine and anserine in the amount 85-97 wt.-% of the native content of these components in poultry muscle tissue, 1-7 weight parts of amino acids, 0.5-12 weight parts of oligopeptides of molecular mass 10 kDa, not above, and 0.1-15 weight parts of cyclic and polycyclic phenolic compounds as measured for 1 weight part of carnosine and anserine in the complex. This complex is prepared by enzymatic hydrolysis of milled and homogenized water muscle tissue in preferable dilution homogenate with water in the range 0.2-0.6 and with using proteolytic enzymes in the amount 2-5 wt.-% of the protein content and working at pH 4.5-8.5 and at enhanced temperature being preferably at 45-65°C. Product is isolated as extract or powder prepared in drying the extract. Invention provides enhancing effectiveness of the claimed complex.
EFFECT: improved method for preparing, valuable properties of complex.
7 cl, 6 tbl, 6 ex
FIELD: medicine, cardiology, gastroenterology.
SUBSTANCE: invention relates to a method for treatment of ulcer-erosion injures in gastroduodenal region in patients with arterial hypertension. Method involves detection of immune disturbances and carrying out the combined immunomodulating therapy and hypotensive therapy. Immunocorrecting complex consists of licopide, cortexinum, vetoronum TK in arterial hypertension of I-II degree and comprises superlymph additionally in arterial hypertension of III degree. Method provides attaining optimal results in treatment for relatively short time due to adequate immunocorrection in such patients.
EFFECT: improved method for treatment.
5 cl, 6 tbl, 2 ex
FIELD: organic chemistry, medicine, pharmacology.
SUBSTANCE: invention relates to new inhibitors of thrombin of the formula (I)
method for their preparing, intermediate compounds used for their preparing of the formula (II)
and a pharmaceutical composition comprising compounds of the formula (I). Invention provides enhancing effectiveness in inhibition of thrombin.
EFFECT: improved preparing method, valuable medicinal properties of compounds.
23 cl, 61 ex
FIELD: medicine, pharmacy.
SUBSTANCE: invention relates to a combined medicinal agent used in treatment of arterial hypertension. The proposed agent comprises the combination of enalapril maleate and hydrochlorothiazide as an active component, and also sodium hydrocarbonate, starch, lactose, iron oxide and stearate as accessory substances. The proposed agent is stable in storage and releases the active component easily.
EFFECT: improved and valuable properties of agent.
8 cl, 1 tbl, 5 ex
FIELD: veterinary science, pharmacy.
SUBSTANCE: invention proposes a composition for antioxidant protection of cells, tissues and a whole body against hyperproduction of free radicals in acute inflammation, chemical thermal and radiation damages. The composition comprises peroxyredoxin Prx VI and, additionally, lipoic acid and pharmaceutically acceptable additives. The composition comprises peroxyredoxin Prx VI and dihydrolipoic acid taken in the effective amount in the ratio peroxyredoxin Prx VI to dihydrolipoic acid in the range (w/w) from 1:1 to 50:1 wherein peroxyredoxin Prx VI can represents human recombinant peroxyredoxin Prx VI. Also, invention relates to a method for enhancing antioxidant protection of mammals involving delivery of indicated pharmaceutical composition is carried out into intercellular space of tissue, organ or a whole body of mammal. The delivery can be carried out by passive or active diffusion in application or spraying, by parenteral or endolumbal administration by injection, by parenteral administration, infusion, inhalation, drainage, by sublingual, vaginal or rectal administration, by nasal or ophthalmic drops. Except for, the delivery can be carried out with using other therapeutic agent, in particular, interferon simultaneously. Invention provides prophylaxis of secondary alternative damages, recovery of epithelial tissue, protection of biomacromolecules against effect of irradiation.
EFFECT: valuable medicinal properties of composition.
6 cl, 9 tbl, 11 dwg, 45 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: present invention refers to crystalline modifications: 1 (polymorphous form F), 2 (polymorphous form I) and 3 (polymorphous form X) of monosodium salt of D-isoglytamyl-D-tryptophan (1:1) characterised by powder X-ray pattern peaks presented in the application materials, as well as to pharmaceutical compositions containing them. The invention describes their use for treating various diseases and body conditions of at least one autoimmune diseases specified in a group consisting of psoriasis, atopic dermatitis and rheumatoid arthritis.
EFFECT: present invention describes the methods for producing the declared crystalline modifications of monosodium salt of D-isoglytamyl-D-tryptophan (1:1).
42 cl, 4 ex, 9 dwg