Peptide composition eliciting immunity regulating property

FIELD: medicine, immunology, peptides.

SUBSTANCE: invention relates to a new composition of biologically active substances. Invention proposes the composition comprising of peptides of the formula: Arg-Gly-Asp and H-Tyr-X-Y-Glu-OH wherein X means Gln and/or Glu; Y means Cys(acm) and/or Cys that elicits ability to inhibit the proliferative response for phytohemagglutinin, to induce the suppressive activity of mononuclear cells and ability of peptides to induce secretion of immunosuppressive cytokines of grouth-transforming factor-β1 and interleukin-10 (IL-10). The composition can be prepared by a simple procedure.

EFFECT: valuable biological properties of composition.

3 cl, 16 tbl, 9 ex

 

The invention relates to medicine, in particular to new biologically active substances (BAS), with immunoregulatory properties, and can be used in practical medicine, veterinary medicine, as well as in experimental biochemistry.

Known copolymer-1 (COP-1), manufactured by TEVA under the name “Copaxone” (see the Monograph authors Eyisi, Tlemen and Annouce “Multiple sclerosis”, Moscow, 1997, page 317).

This drug has a wide functionality, however, getting it is very complex because of the high molecular weight, which increases its production.

Renowned pharmaceutical composition used for treatment of multiple sclerosis by neutralizing antimelanoma antibodies (see RF patent №2121850, class a 61 L 38/10 from 22.10.91 year).

However, the known arrangement does not possess immunoregulatory properties.

The technical result is the development of a new synthetic composition having immunoregulatory capabilities and ease of access.

To solve this technical problem is proposed composition with immunoregulatory properties, comprising at least two peptide of the formula Arg-Gly-Asp and H-Tyr-X-Y-Glu-OH, where X is Glu and/or Gln, Y - Cys and/or Cys(acm), and the peptides can be taken in equal shares or in shares, zootoxin the e which is 1:4, respectively.

The invention consists in that the use of compositions with the above formula allows us to provide extended functionality effects on mononuclear cells by the ability to induce high-level secretion of immunosuppressive cytokines.

In addition, low molecular weight components of the proposed composition determines the ease of its manufacture.

The comparison of the proposed composition with the closest analogue suggests the criterion of “novelty”, and the lack of analogues distinctive signs under the criterion of “inventive step”.

Preliminary tests suggest about the possibility of wide industrial production and use in medicine.

Obtaining the claimed composition can be carried out by known methods of peptide synthesis.

Peptides comprising the inventive composition according to the present invention, can be obtained in accordance with conventional and well known methods of synthesis of polypeptides. For the synthesis of the used activated amino acids with protected functional groups (company: Sigma, Merk).

Synthesis of peptides was carried out by generally accepted methods of solid-phase synthesis: in particular, Fmoc-scheme of formation of the peptide bond, to the which involves the use of temporary masking of alpha-amino groups fluorenylmethoxycarbonyl (Fmoc) group, stable in the acidic environment, but tsepliaeva bases. The first peptide RGD-arginyl-glycyl-aspartic acid (Arg-Gly-Asp) was synthesized in the form of the free acid in RAS polymer company MilliGen/Biosearch (USA) attached to it starting C-terminal amino acid residue Fmoc-aspartic acid. The condensation reaction was carried out using 1-oxibendazole or pentafluorophenyl esters, consistently using the corresponding esters of glycine and arginine. Completeness of passage of the reaction was monitored using the ninhydrin test. In the case of incomplete passing the reaction they were re-condensation. For cleavage of the peptide from the polymer used a mixture triperoxonane acid (TN) and phenol (95/5). Purification of the peptide was performed by gelfiltration on Sephadex G-10 and characterized data quantitative amino acid analysis, analytical reversed-phase high-performance liquid chromatography and thin-layer chromatography.

The synthesis of the second peptide of the proposed composition was also carried out on solid phase. When this was applied stiroldivinilbenzol resin to which type of education benzyl ether was added glutamic acid, using N-Z-1-glutamate-α-N-hydroxysuccinimide ether, followed by treatment of the compound N-t-BOC-S-atsetamidometil-1-cysteine in Pris the accordance of DICYCLOHEXYL carbodiimide in dioxane, with the subsequent laundering of solid-phase polymer, removing the BOC-protective group and formed of DICYCLOHEXYL carboxyrhodamine.

Next to solid phase polymer was added N-CBZ-1-glutamine-α-N-hydroxysuccinimide ether, and in the final stage after removal of the CBZ protecting group was treated with N-t-BOC-1-tyrosine-N-hydroxysuccinimide ether. After removal of the BOC protective group carried out the removal of the formed peptide by alkaline hydrolysis.

The second peptide and recrystallized liofilizirovanny. The presence and sequence of amino acids, as well as the purity of the peptide was checked by thin layer chromatography, using as a single peptide, and the products of its partial or complete hydrolysis. Received second peptide 1 type corresponded to the formula: H-Tyr-Gln-Cys(acm)-Glu-OH, where AFM atsetamidometil, the protective group for the sulfur of cysteine, which was not removed because it spontaneously is easily removed in the conditions of the body and in cell cultures, resulting in the effective agent is a peptide with the formula H-Tyr-Gln-Cys(asm)-Glu-OH.

N - the beginning of a peptide; a Tight - amino acid residue tyrosine; Gln - amino acid residue glutamine; Cys is the amino acid cysteine residue, Glu - glutamic acid (glutamate) or its remainder, IT is the end of the peptide.

For synthesis of the second peptide of the 2nd type with the formula H-Tyr-Glu-Cys(acm)-Glu-OH was used that W is the principle of replacing them with the corresponding step of adding N-CBZ-glutamine-N-hydroxysuccinimide ether in N-Z-glutamate-α -N-hydroxysuccinimide ether. All reagents and activated amino acids with protected groups were brand (Sigma, Merk).

Treating the peptide with the above formula in the presence of ions of mercury removed acetamidomethyl group, freeing the SH group of cysteine. Thus obtain peptides with formula without (asm).

Upon receipt of a composition according to claim 2 of the formula mix the first peptide with a second peptide of each type separately in the proportion of 1:4, respectively.

Upon receipt of a composition according to claim 3 of the formula mix the first peptide with a second peptide of each type separately in equal shares.

The result can be obtained compositions of four different types. In addition, to obtain a composition of other types used instead of X at the second peptide is simultaneously a mixture of Gln and Glu. It should be noted that carried out the preparation of the composition of the two peptides in various unequal shares. However, the benefits of obtaining a composition before composition with peptides in fractions, the ratio of which is 1:4, was not observed.

The biological activity of the inventive compositions of each of the four types was determined as follows.

Example 1

1. Studied the effect of a composition consisting of two peptides with formulas Arg-Gly-Asp and H-Tyr-X-Y-Glu-OH, where X is Glu, Y - Cys(acm)on the proliferative activity of mononuclear is peripheral blood cells, selected by the method of Voim 1969

The range of the used dose was 0.03 to about 3.00 mg/ml

The research results are reflected in table 1.

Dose in mcg/ml0,020,030,080,150,30,61,22,43,03,6
Index Inhib %05172433453624120

The results showed that the composition inhibits the proliferative activity of mononuclear peripheral blood cells caused by phytohemagglutinin (PHA).

2. Investigated the influence of the composition on the ability to induce suppressor activity of mononuclear cells in peripheral blood.

At the first stage receives the suspension OLS method sedimentation of cells in a single-stage gradient ficoll-protracta (method Voim).

Peripheral blood is collected from the donor by venipuncture at the same time and placed in a test tube with a solution of heparin per 1 ml of blood 20-30 units of heparin. Next, the blood is diluted with Hanks solution without CA++and Mg++in the ratio of 1:2 and layer on the gradient ficoll-eurotrust (density 1,078).

For the eat spend the centrifugation mode 400 g for 30 minutes Suspended OOC out of interphase transferred into a centrifuge tube, add the Hanks solution without CA++and Mg++and produce 3 consecutive centrifugation for 10 min to launder cells from solution ficoll-eurotrust. After the 3rd centrifugation the precipitate OLS resuspension in 1 ml of medium 199 and count the number of mononuclear cells with the camera Goryaeva.

In the second stage OLS divided into two equal parts, the first of which is cultivated without activator suppressor, the second - with activator suppressor, which is used as the analyzed composition consisting of 2 peptides.

MNCs were cultured in penicillin vials closed with rubber corks No. 14,5 at a temperature of 37°C. the Environment of the cultivation of RPM1-1640 with the addition of 20% serum IV AB blood group and glutamine.

In each vial cultivate 5×106cells in 2.0 ml of complete medium. To culture for induction of suppressor type composition of the 2 peptides at doses of 0.03 to 3.0 mg/ml

The cell cultivation carried out 48 hours After that MNCs were washed off the environment of the cultivation and are blocking proliferation by treatment with mitomycin C - 40 µg/ml for 30 min at 37°C. Then washed with medium 199 with 5% serum IV AB (chilled) three times. Sediment cells resuspension, count the number of nucleated cells, determine Aut the percentage viability of the cells with 0.1%Trypanosoma blue solution and diluted to the required concentration. In this case, all operations are done separately with the control cells and stimulated with compositions, and to launder cells use silikonizirovannaya the dishes.

In the next step, each part of the control and stimulated composition OOC add freshly isolated MNCs stimulated phytohemagglutinin (PHA), which are responsible of the test cells in equal proportions (0,5×106:0,5×106cells/ml) to obtain the test cultures. The cultivation is carried out within 72 hours After using the N3-thymidine assess the proliferation of the test cultures and the amount of suppression is judged by the degree of reduction of proliferation in them. Index suppression (IP) is determined by the formula:

The results of studies of the effect of the composition consisting of two peptides with formulas Arg-Gly-Asp and H-Tyr-X-Y-Glu-OH, where X is Glu, Y - Cys(acm), the ability to induce suppressor activity MTL reflected in table 2.

The dose range of 0.02-3.6 µg/ml.

Table 2.
Dose in mcg/ml0,020,030,080,150,30,61,22,43,03,6
Index PMPs. %02 182543281450

It is established that the composition in doses of 0.03 µg/ml of 3.00 μg/ml induces suppressor activity of mononuclear cells in peripheral blood.

Example 2.

In the study of the composition of the 2nd type (Arg-Gly-Asp and H-Tyr-X-Y-Glu-OH, where X is Gln, Y - ys(acm), mimicked the actions of paragraphs 1 and 2 of example 1.

The results of the studies are shown in tables 3 and 4.

Table 3.

The effect of the composition consisting of two peptides of formulas (Arg-Gly-Asp and H-Tyr-X-Y-Glu-OH, where X is Gln, Y - Cys(acm) on the proliferative activity of MNCs.
Dose in mcg/ml0,020,030,080,150,30,61,22,43,03,6
Index Inhib %0516212844291870

Table 4.

The influence of the composition consisting of two peptides of formulas (H-Arg-Gly-Asp-OH and H-Tyr-X-Y-Glu-OH, where X is Gln, Y - Cys(acm) on the ability to induce suppressor activity of MNCs.
Dose in mcg/ml0,020,030,080,150,30,61,22,43,03,6
Index PMPs. %0315243239231240

The result was that the song also inhibits the proliferative activity of mononuclear peripheral blood cells caused by the PHA, and induces suppressor activity of mononuclear peripheral blood cells when used in doses of 0.03 µg/ml of 3.00 mg/ml

Example 3.

In the study of the action of the composition of the 3rd and 4th types of 2 peptides with Arg-Gly-Asp and H-Tyr-Gln-Cys-Glu-OH and Arg-Gly-Asp and N-Tight-Glu-Cys(acm)-Glu-OH, taken in equal parts and in the ratio of 1:4, repeated action item 1 and item 2 of example 1.

The results of the action research of the composition of the two peptides with Arg-Gly-Asp and H-Tyr-Gln-Cys-Glu-OH on proliferative activity and ability to induce suppressor activity of the OLS regression are shown in tables 5 and 6.

The dose range of 0.02-3.6 µg/ml.

Table 5.
Dose in mcg/ml0,020,030,080,150,30,6 1,22,43,03,6
Index Inhib %04183037433122110

Table 6.
Dose in mcg/ml0,020,030,080,150,30,61,22,43,03,6
Index PMPs. %0411182541261130

The results of the action research of the composition of the two peptides with Arg-Gly-Asp and H-Tyr-Glu-Cys(acm)-Glu-OH on proliferative activity and ability to induce suppressor activity of the OLS regression are shown in tables 7 and 8.

The dose range of 0.02-3.6 µg/ml.

30
Table 7.
Dose in mcg/ml0,020,030,080,150,30,61,22,43,03,6
Index Inhib %061435462918120

Table 8.
Dose in mcg/ml0,020,030,080,150,30,61,22,43,03,6
Index PMPs. %0410173149332150

The result was that these compositions inhibit the proliferative activity of mononuclear peripheral blood cells caused by the PHA, and induce suppressor activity of mononuclear peripheral blood cells when used in doses of 0.03 µg/ml of 3.00 mg/ml

In addition, we investigated other types (variants) of the claimed composition, consisting of 2 peptides, which also confirmed their ability to inhibit the proliferative activity and to induce suppressor activity of mononuclear cells in human peripheral blood. It should be noted the considerable ease of access to all variants of the compositions of the 2 peptides.

Example 4

Investigated the influence of the composition consisting of 2 peptides of the formula is e (Arg-Gly-Asp and H-Tyr-X-Y-Glu-OH, where X is Gln and/or Glu, Y - Cys(acm) and/or Cys on the ability to induce the production of cytokines, transforming growth factor beta 1 (TGF-β1) and interleukin-10 (IL-10) mononuclear cells (MNCs) in the peripheral blood.

In the first phase have received a suspension OLS method sedimentation of cells in a single-stage gradient ficoll-ultratrust (method Voim).

MNCs were divided into 2 equal parts, the first of which were cultivated without composition consisting of 2 peptides, one with a composition consisting of 2 peptides.

MNCs were cultured in penicillin vials closed with rubber corks No. 14,5 at a temperature of 37°C. the cultural Medium RPMI-1640 with the addition of 20% serum IV AB blood group and glutamine.

In each vial were cultured 5×106MNCs in 1 ml of complete medium. The culture of MNCs to induce the production of cytokines TGF-β1 and IL-10 were added to a composition consisting of 2 peptides.

The cultivation was carried out for 24 hours. Then the culture medium was separated from cells by centrifugation 1000 g for 10 minutes and it took 500 ál needed to perform the analysis.

When determining the amount of TGF-β1 in the culture medium was originally produced by extraction of the samples. This stage of analysis allows the release of TGF-β1 package, making it available for analysis.

For this purpose polypropylene PP is Lenovo tube made of 0.25 ml (250 μl) of culture medium and 0.05 (50 ál) of the extracting solution. Mixed contents of the vortex. Incubated for 30 minutes at 4°C. was Added in a test tube 250 μl working buffer solution for dilution of standards. At this stage, the dilution of culture medium 2.2 times.

Then took required for analysis, the number 8 hole strips of collapsible microplate coated with antibodies to TGF-β1.

To improve the reliability of the results of the analysis and control samples were placed in duplicate, using for each sample two holes. Contributed by 200 µl of each standard and extracted sample or cotrona into the appropriate wells. Added 500 μl of biotinylated antibodies to TGF-β1 all wells were mixed by tapping on the tablet. Covered the plate with foil and incubated for 30 minutes at room temperature. Completely delete the contents of the wells. Washed all the wells 4 times with working buffer solution, adding it each time with 400 ml to all wells. Each time was completely filled and removed from the wells strips solution. After washing away excess moisture released tapping the inverted stripe on filter paper. Added 100 ál of working strength conjugate streptavidin-peroxidase to each well, except chromogenic Blanca. Covered the plate with foil and incubated for 30 mi the ut at room temperature. Completely delete the contents of the wells. Washed the wells 4 times with working buffer solution. Next was introduced into each well, 100 μl of chromogenic solution of tetramethylbenzidine (TMB). Incubated for 20-30 minutes at room temperature in the dark until the blue color in the wells with the calibration breakdown with a maximum content of TGF-β1.

Contributed to each well 100 ál of stop solution (a solution of 1N sulfuric acid) to stop the enzymatic reaction. They mixed the reagents by gently tapping the strip holder. The color of the solution changed from blue to yellow.

The registration results were photometrically on the photometer for assay at a wavelength of 450 nm immediately after stopping the enzymatic reaction.

To determine the concentration of TGF-β1 in the samples were building a calibration curve in the coordinates: the x - axis the concentration of TGF-β1 in the calibration samples (WG/ml); y-axis is the corresponding value of optical density.

By means of a calibration curve based on the obtained values of optical density was determined the concentration of TGF-β1 in the samples. If the samples were pre-diluted, obtained values of the concentrations were multiplied by the dilution factor (10, 100,1000 and so on).

To determine the amount of IL-10 in the culture media who took required for analysis, the number 8 hole strips of collapsible microplate, covered with antibodies to IL-10.

To improve the reliability of the results of the analysis and control samples were placed in duplicate, using for each sample two holes.

Made of 50 µl of each standard, test sample or control sample into the appropriate wells. Added 50 μl of incubation buffer to the wells containing standards, and 50 µl of the dilution of the working solution in the wells containing culture medium. Closed the tablet film and incubated 2 hours at room temperature. Completely delete the contents of the wells.

Washed all the wells 4 times with working buffer solution, adding it each time in 400 ál to all wells. Each time was completely filled and removed from the wells strips solution. Added 100 μl of biotinylated antibodies to IL-10 in all wells were mixed by tapping on the tablet.

Closed the tablet film and incubated 2 hours at room temperature. Completely delete the contents of the wells. Washed all the wells 4 times with working buffer solution. Added 100 ál of working strength conjugate streptavidin-peroxidase to each well except chromogenic form.

Closed the tablet film and incubated for 30 minutes at room temperature. Completely delete the contents of the wells. Washed the wells 4 times with working buffer solution./p>

Next was introduced into each well, 100 μl of chromogenic solution of tetramethylbenzidine (TMB).

Incubated for 20-30 minutes at room temperature in the dark until the blue color in the wells with the calibration breakdown with the highest levels of IL-10.

Contributed to each well 100 ál of stop solution (a solution of 1N sulfuric acid) to stop the enzymatic reaction. They mixed the reagents by gently tapping the strip holder. The color of the solution changed from blue to yellow.

Check results also carried out photometrically on the photometer for assay at a wavelength of 450 nm immediately after stopping the enzymatic reaction.

To determine the concentration of IL-10 in the samples were building a calibration curve in the coordinates: the x - axis the concentration of IL-10 in the calibration samples (WG/ml); y-axis is the corresponding value of optical density.

By means of a calibration curve based on the obtained values of optical density was determined the concentration of IL-10 in the samples. If the samples were pre-diluted, obtained values of the concentrations must be multiplied by the dilution factor (10, 100, 1000 and so on).

As a result of the research it was found that a composition consisting of 2 peptides, has the ability to induce PR the products at the same time 2 cytokines - TGF-β1 and IL-10 by mononuclear cells of peripheral blood.

Example 5-1

The effect of the composition consisting of two peptides with formulas Arg-Gly-Asp and H-Tyr-X-Y-Glu-OH, where X is Glu, Y - Cys (acm)on the proliferative activity of MNCs with a ratio of the first peptide with a second 4:1.

The range of the used dose was 0.02 μg/ml to 3.6 mcg/ml

The research results are reflected in table 9.

Table 9.
Dose in mcg/ml0,020,030,080,150,30,61,22,43,03,6
Index Inhib %0412212841271140

Example 5-2.

The results of studies of the effect of the composition consisting of two peptides with formulas Arg-Gly-Asp and H-Tyr-X-Y-Glu-OH, where X is Glu, Y - Cys(acm), the ability to induce suppressor activity MNF when the ratio of the first peptide with a second peptide 4:1 is presented in table 10. The dose range of 0.02-3.6 µg/ml.

Table 10.
Dose in mcg/ml0,020,030,08 0,150,30,61,22,43,03,6
Index PMPs. %037152639251140

Example 6-1 and 6-2.

The results of the action research of the composition consisting of two peptides with formulas Arg-Gly-Asp and H-Tyr-X-Y-Glu-OH, where X is Gln, Y - Cys(acm)on proliferative activity and ability to induce suppressor activity of MNCs with a ratio of the first peptide with a second peptide 10:1 are shown in tables 11 and 12.

The dose range of 0.02-3.6 µg/ml.

Table 11.
Dose in mcg/ml0,020,030,080,150,30,61,22,43,03,6
Index Inhib %0712172229201050

0,6
Table 12.
Dose in mcg/ml0,020,030,080,150,31,22,43,03,6
Index PMPs. %0612182332201150

Example 7-1 and 7-2.

The results of the action research of the composition of the two peptides with Arg-Gly-Asp and H-Tyr-Gln-Cys-Glu-OH on proliferative activity and ability to induce suppressor activity of MNCs with a ratio of the first peptide with a second peptide 1:4 are shown in tables 13 and 14.

The dose range of 0.02-3.6 µg/ml.

Table 13.
Dose in mcg/ml0,020,030,080,150,30,61,22,43,03,6
Index Inhib %0716222839251960

Table 14.
Dose in mcg/ml0,020,030,080,150,30,61,22,43,0 3,6
Index PMPs. %039192638241240

Example 8-1 and 8-2.

The results of the action research of the composition of the two peptides with Arg-Gly-Asp and H-Tyr-Glu-Cys(acm)-Glu-OH on proliferative activity and ability to induce suppressor activity of MNCs with a ratio of the first peptide with a second peptide of 1:10 are shown in tables 15 and 16. The dose range of 0.02-3.6 µg/ml.

Table 15.
Dose in mcg/ml0,020,030,080,150,30,61,22,43,03,6
Index Inhib %0510182431211260

Table 16.
Dose in mcg/ml0,020,030,080,150,30,61,22,43,03,6
Index PMPs. %0 37122229231660

Thus examples 5-1, 5-2, 6-1, 6-2, 7-1, 7-2, 8-1, 8-2, in which the peptides were mixed in the ratio 4:1, 10:1, 1:4 and 1:10, indicate the absence of benefits of getting these songs before composition with peptides in fractions, the ratio of which is 1:1.

Example 9.

Investigated the influence of the composition of all the above types, consisting of 2 peptides, on the survival of skin allograft.

Were taken 2 groups of nonlinear mice - one control, the other experienced. All mice were dissected flap 1.5×2 cm and was transferred to another mouse, which also were excised skin of the same size. All operations were performed under conditions of asepsis (sterile suture material, tools, etc). Skin flap to engraftment was treated in a solution of antiseptics. Drug (composition of 2 peptides) was administered in sterile conditions. The room in which there was a mouse in the postoperative period, two times per day processed germicidal lamp (animals in this period was in the other room).

The first group of mice after transplantation of skin area received injections of saline control mice. Another group of mice poltransplant skin were subcutaneously injected with 1 time per day of the composition, consisting of 2 peptides, daily for 10 days. In the control group, starting from 4-to 8-day dermal graft was wrinkled, drying up, and on the 10th day came the rejection (one mouse on day 6). In the experimental group the skin graft has taken root and rejection came only 30 days.

Thus, the introduction of the composition of all the above types, consisting of 2 peptides, experienced mice, which were allotransplantation skin contributes to the prolongation of graft survival.

To study the security of the claimed composition of the 2 peptides was performed to study its acute toxicity. Study of acute toxicity was carried out in accordance with the Methodological recommendations of the Pharmacological Committee of the Russian Federation “Requirements for preclinical study obmotochnogo action of new pharmacological substances”, Moscow, 2001, the results of the study showed that intraperitoneal injection of 1000 times the dose of the composition there were no acute toxic effect, and at these doses has not been possible to reach his LD50. Thus, the proposed composition consisting of 2 peptides, is not toxic.

According to the present invention, all types of compositions of the formula Arg-Gly-Asp and H-Tyr-X-Y-Glu-OH, where X is Glu and/or Gln, Y - Cys and/or Cys(acm)have biological activity, describe the Noi in the examples and can find application in medicine and biology.

Although the above invention has been described in detail to illustrate, for the average person skilled in the art will be quite clear, in light of the description of this invention that can be made certain changes and modifications of the invention without departing from the idea or the scope of the proposed claims.

1. The composition having immunoregulatory properties, comprising at least two peptide of the formula Arg-Gly-Asp and N-Tight-X-Y-Glu-OH, where X is Gln and/or Glu, Y - Cys(acm) and/or Cys.

2. The composition according to claim 1, characterized in that the peptides are taken in fractions, the ratio of which is 1:4, respectively.

3. The composition according to claim 1, characterized in that the peptides are taken in equal parts.



 

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FIELD: medicine, phytotherapy, pharmacy.

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2 cl, 3 tbl, 1 ex

FIELD: medicine, obstetrics, gynecology.

SUBSTANCE: at the background of therapy conducted one should introduce derinate immunomodulator into the body of pregnant woman additionally nasally per 1-2 drops of 0.25%-solution into each nasal canal 5-8 times daily for 3-5 d and - parenterally per 5.0 ml of 1.5%-solution once daily for 3-8 d along with preparation that improves microcirculation and along with antioxidant at a certain sequence, moreover, derinate should be introduced 30-40 min after application of microcirculation-improving preparation, and antioxidant - 20-30 min after derinate's introduction. The present innovation favors decreased edemas, decreased body weight, stabilization of Macluer-Aldrich test that in its turn enables to avoid perinatal losses, decrease the risk for the development of fetoplacental insufficiency and intrauterine fetal infection.

EFFECT: higher efficiency of therapy.

1 ex, 2 tbl

The invention relates to biopharmacology and comes to obtain the aimed at restoring metabolic processes and correction functions of the immune system

Immunomodulator // 2240794
The invention relates to new biologically active substances from the class alkenylboronic acids and can be used in medicine and biology as the basis for making medicines

Currently, in the treatment of infectious-inflammatory, autoimmune and oncological diseases in clinical medicine is widely used medicines, which have immunomodulatory properties (I. B. Mikhailov

The invention relates to the pharmaceutical industry and relates to the connection of a number of oxyindole, namely ethyl ester of 6-bromo-4-dimethylaminomethyl-1-methyl-5-hydroxy-2-phenylthio-methylindolin-3-carboxylic acid hydrochloride, which is an active substance of the drug Arbidol, and the drug based on it in the form of gelatin capsules

The invention relates to new biologically active compounds, namely diglyceride glycyrrhizic acid methyl ester L-valine of formula (I), stimulating primary immune response

The invention relates to medicine

The invention relates to N-substituted indole-3-glycinamide General formula I, possess Antiasthmatic, antiallergic and immunosuppressive/immunomodulatory action

where R is hydrogen, (C1-C6)alkyl, and the alkyl group optionally contains one phenyl substituent, which, in turn, optionally contains at least one Deputy, selected from the group comprising halogen, methoxy, ethoxy, (C1-C6)alkyl; R1means phenyl cycle containing at least one Deputy, selected from the group comprising (C1-C6)alkoxy, hydroxy, nitro, (C1-C6)alkoxycarbonyl one or fluorine, or R1represents the balance of the pyridine of the formula II

where the carbon atoms 2, 3 and 4 of the remaining pyridine optionally have the same or different substituents R5and R6and R5and R6denote (C1-C6)alkyl or halogen, or R1presents arylamination-2-methylprop-1-ilen group, or R and R1together with the nitrogen atom to which IGN="ABSMIDDLE">

where R7denotes phenyl or pyridinyl; R2means (C1-C6)alkyl, which optionally contains a phenyl residue, which, in turn, optionally substituted with halogen, methoxy group or ethoxypropane, or related to R2(C1-C6)alkyl group optionally substituted 2-, 3 - or 4-pyridinium residue; R3and R4are the same or different substituents and represent hydrogen, hydroxy, (C1-C6)alkoxy, (C1-C3)alkoxycarbonyl or (C1-C3)alkoxycarbonyl(C1-C3)alkyl, or R3is cyclopentanecarbonitrile; Z denotes Oh, and alkyl, alkoxy or alkylamino mean as an unbranched group, such as methyl, ethyl, n-propyl, n-butyl, n-hexyl and branched alkyl groups such as isopropyl or tert-butylene group; halogen means fluorine, chlorine, bromine or iodine and alkoxygroup means methoxy, propoxy, butoxy, isopropoxy, isobutoxy or phenoxypropan, and their pharmaceutically acceptable salts with acids

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