Peptides with organizaitnal activity, pharmacologically active composition

 

(57) Abstract:

Usage: in the chemistry of biologically active peptides with organizaitnal activity. Disclosed are peptides of formula I,

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where Xaa is a neutral aliphatic amino acid, Yaa - essential amino acid, Zaa - amino acid character, and at least one of the residues Xaa and Zaa can be skipped and pharmacological composition based on them. 2 C. and 14 C. p. F.-ly, 16 ill., table 1.

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

Biologically active protein with organizaitnal activity, which is extracted from the body of the animal or human and is called HRV (protecting the body connection), described in European patent 0432400, and also published in P. Siciric et. al., Exp. Clin. Gastroenterol, I, 15-26, 1991. Protein has a mol. a mass of about 400005000, its structure is defined only in part. This compound has a biological activity of a wide range of actions: protects against ulcers, has hepatoprotective, anti-virus, anti-edema, a common anti-inflammatory, anticancer and lectern of the nervous system and disorders of dopaminergic etiology, surgery, dentistry, restoring the ability to procreate and in veterinary medicine. But such a broad spectrum of activity may be due to only partially certain patterns or even insufficient purity and homogeneity of selected compounds HRV.

In accordance with one aspect of the present invention proposed a class of synthetic peptides with unexpectedly high biological activity, especially when the protection of a living organism. In any case, these synthetic compounds with well-established structure a lot better than high molecular weight protein HRV with a partially installed structure, which can be obtained only by the difficult way of the dubious nature of the source.

More specifically, this invention relates to a new class of very biologically active peptides containing 8 to 15 amino acid residues. Peptides represented by the following basic structural formula (used three-letter amino acid designations, and the number under amino acid residues correspond to the position of the residue in the peptide chain)

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Values of the substituents Xaa, Yaa and Zaa are given in the table.

Predoc (abbreviation: th. ID. N 1).

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PEFC. ID. N 2

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PEFC. ID. N 3

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In accordance with another aspect of the present invention proposed a similar peptides that are in the C-terminal position are aminogroup or carboxypropyl. These peptides have the above structural formulas, in which at least one and no more than seven amino acid residues in positions 1-15 are missing and at least one of the substituents Xaa, Yaa and Zaa has the meaning specified in the table.

Preferred peptides of the following formula:

PEFC. ID. N 4

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PEFC. ID. N 5

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PEFC. ID. N 6

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PEFC. ID. N 7

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PEFC. ID. N 8

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In accordance with another aspect of the invention, the peptides, which have the above structural formula and in which at least one amino acid residue is absent and at least one other amino acid residues substituted, as indicated in the table, transformed into a circular form by formation of a new communications CO-NH between the first and last amino acid residues in the molecule.

Preferred the following peptides:

PEFC. ID. N 9

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PEFC. ID. N 10

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It was found that described in the national Assembly, or in excess thereof.

Pharmacological studies described peptides was performed on a variety of normal samples "in vitro" and "in vivo". Found the following pharmacological properties:

1)a Protective effect on the liver and pancreas

Protective effect of newly synthesized peptides having a pet. ID. N 2, 3, 4, 6, comparisons with action standards (bromocriptine, draft, somatostatin) in various experimental models (rats with a damaged liver and pancreas). Damage reached 24-hour legirovaniem bile ducts, 24-hour legirovaniem bile duct + hepatic artery, 48 h, the limiting stress, applique CCl4. Applied peptide, administered in an amount of from 10 μg to 10 ng/kg of body weight intragastrically or intraperitoneally significantly prevented necrosis of the liver and pancreas, as well as fatty degeneration in animals subjected to 24-hour legirovanie bile ducts, 24-hour legirovanie bile duct + hepatic artery, 48 h limiting stress, appliques CCl4. Biological value content of bilirubin, SGOT, SGPT are fully in accordance with the specified macro/microscopic data used Wistar albino rats with a mass of 190-250, Rats was conducted unilateral nephrectomy, resulting in an increase in the mass of the remaining kidney from a group of control animals and groups of animals, which were injected intraperitoneally peptides with PEFC. ID. N 2, 3, 4, 6 in the amount of 10 μg/kg of body weight one hour prior to surgery. In groups of animals, which were injected peptides were observed decrease increase mass remaining kidney.

Were compared biochemical parameters in both groups. It turned out that all applicable peptides improve the function of the remaining kidney.

b) the Use of gentamicin.

In our experiments we used female Wistar albino rats. Damage to the tubules induced by gentamicin, administered at 40 mg/kg once a day for 30 days. All animals were exposed 5 days after the last injection of drugs. Significant damage to tubules was observed in the control group, whereas smaller microscopic violations installed in all groups of animals, which were introduced above peptides.

(c) the Use of chloride of mercury.

The experiments were performed on white Wistar rats. The mercury chloride was injected intravenously at 2 mg/kg of body weight. Peptides with PEFC. ID. N 2, 3, 4, 6 enter scopacasa the results showed a protective effect in animal groups, which introduced these peptides.

3) Effect on testicular damage

The effect of peptides with PEFC. ID. N 2, 3, 4, 6 damaged testicles examined by introduction of ketoconazole (24 mg/kg, intragastrically), testosterone (30 mg/kg, intraperitoneally) or 6Gy-irradiation. One hour prior to the introduction of such compounds or irradiation they were introduced peptides with PEFC. ID. N 2, 3, 4, 6. These peptides showed a protective effect.

4) Effects on reproduction/commodity livestock.

a) Peptides with PEFC. ID. N 2, 3, 4, 6, and reproduction. Impact on oligoasthenospermia.

Studies were performed on 10 male age 30-40 years with a diagnosis of oligoasthenospermia. The ejaculate was taken from these patients after 3-4 days after abstinence. After dissolution (30 min) to 0.5 ml of semen was added to 0.5 ml of medium. The control medium containing HAM-F10 with 10% deactivated hordley serum. Experimental medium containing an additional 2 or 4 mcg/ml HRV. After distribution for 90 min at 37oC in an atmosphere containing 5% CO2in the cell reproduction Horwell to determine the number of moving, moving in place and immobile spermatozoa in 1 ml of ejaculate. Preliminary results showed no influence on the deed) there was a significant impact.

b) Peptides with PEFC. ID. N 2, 3, 4, 6, and reproduction.

The action of these peptides was studied in mice with the previous three pregnancies. 20 days after the termination of the last lactation animals were subjected to copulation. The peptides were administered at 10 mg/kg of body weight 1 time per day during the entire gestation period (19-21 day) and lactation (week 3). The control group received simultaneously equal volume of saline. Registered number of offspring per female and a lot of offspring.

A thorough statistical analysis showed a significant increase in the number of offspring per female, and as a surprise, no differences in the mass of the bodies in the studied time intervals. All females were left to recover for the next 25 days after the cessation of lactation. Then again Coulibaly to investigate the impact of the fifth pregnancy. High speed reproduction with a high potential for lactation relative to control group was observed in all groups of animals, which have introduced these peptides.

It should be noted that these peptides could improve the reproductive performance in very old mice. In conclusion, it must register in vitro using human sperm.

c) Commodity livestock.

1419 healthy piglets (4306) received 10 μg of peptides with PEFC. ID. N 2, 3, 4, 6 immediately after birth, others 1440 pigs - on the 13th day of life 1 hour before castration, the rest 1477 received only conventional Fe-therapy. Bruises, culling, mortality, body weight and feed consumption were evaluated after 4 weeks. Applied peptides, entered one at a time, significantly reduced the rate of culling (in the group which was administered peptides immediately after birth) and the rate of injury (in all groups of animals, which introduced the peptides). In animals, which introduced the peptides marked with the same bodyweight in a significant reduction of feed consumption. Impact on infection: investigated forty-28-day, separated from the uterus of pigs infected with enterocolitis E. coli. The peptides were administered 20 animals (10 μg/kg of body weight, mimisan). One month it was observed a significant reduction in mortality in animals that were injected peptides.

In another experiment at 1200 healthy piglets examined the mortality rate after the first week. 400 animals received these peptides immediately after birth in addition to the conventional Fe-therapy. In groups of animals, which introduced these peptides (10 µg/kg body mass, m is recommend that the effect of peptides with PEFC. ID. N 2, 3, 4, 6, entered in the amount of 10 μg or 10 ng/kg body weight intraperitoneally for 5 minutes before injection of amphetamine or simultaneously with it. In groups of animals, which were injected peptides, a significant reduction of behavior problems caused by amphetamine.

6) Effect on bleeding time.

The effect of peptides with PEFC. ID. N 2, 3, 4, 6 explored the usual tests for bleeding circumcision tail or inside) in mice and rats with the use of heparin (1000 M. E. subcutaneously three hours before bleeding) or without it. The studied peptides (10 μg or 10 ng/kg body weight, intraperitoneally) was administered simultaneously with the induction of bleeding. In all animals, which introduced the studied peptides, found a significant decrease in bleeding time.

7) Impact on the consequences of castration.

For the research was developed a modified technique Allen-Doisy. Animals were injected with different doses (10 μg or 10 ng/kg, intraperitoneally) peptide in a disposable or daily injection mode, both before and after being cut. In groups of animals that were administered the analyzed peptides, watched the prevention or reversion of the various consequences of castration (for example vaginal epithelial what iron occlusion of the renal artery. 10 days after induction the animals were injected peptides with PEFC. ID. N 2, 3, 4, 6 (10 μg or 10 ng/kg body weight, intraperitoneally) or saline, using different modes of administration (single or continuous). In groups of animals that were administered the studied peptides observed a significant reduction in quantities to elevated blood pressure.

9) Effect on experimental diabetes

To study the effect of peptides on induced streptozotocin or alloxan diabetes used male Wistar albino rats having a body weight 175-230, Peptides with PEFC. ID. N 2, 3, 4, 6, entered in quantity of 10 or 100 μg/kg, significantly delayed the onset of diabetes in animals, which introduced streptozotocin. In addition, they significantly prolonged survival time of animals.

10) Impact on thyroparathyroidectomy.

Spent a total destruction of the parathyroid glands in rats Wistar. Then they were injected peptides with PEFC. ID. N 2, 3, 4, 6 (10 ng/kg body weight, intraperitoneally) or saline (5 ml/kg body weight intraperitoneally) using different modes of administration (once or repeatedly). The groups of animals that have entered the studied peptides, the effects of the fair were euthanized after 3 h after ligation of both carotid arteries. One hour before all the animals were injected peptides with PEFC. ID. N 2, 3, 4, 6 (10 ng/kg body weight, intraperitoneally) or saline (5 ml/kg body weight, intraperitoneally). The group of animals who introduced the peptides observed less brain swelling.

12) the Effect on the adrenal glands.

To induce damage to the adrenal animals were treated with aniline (300 mg/kg body weight, subcutaneously for 7 days). At the same time all animals were injected peptides with PEFC. ID. N 2, 3, 4, 6 (10 ág/kg 10 ng/kg body weight, intraperitoneally) or saline (5 ml/kg body weight, intraperitoneally). Measured weight of the adrenal glands and microscopy was determined changes. The analyzed peptides significantly reduced aniline-induced increase in adrenal masses (with both doses) compared to the control group. Protective effect of the peptides on the adrenal depended on the dose of the peptides. Since it is generally accepted that induced by aniline disorders associated with hypersecretion of adrenocorticotropic hormone(ACTH), it is possible to establish a causal relationship between the studied peptides and ACTH. This position can also confirm the detection of the same body mass in a group of animals who introduced the peptides and the control of the 48 h after pretreatment with them studied peptides (10 μg or 10 ng/kg of body weight, intraperitoneally/intragastrically). Reduction of elevated adrenal masses in animals entered the studied peptides were aligned with the corresponding microscopic observations.

13) the Effect on the temperature of the violation.

The increased body temperature caused by the introduction of yeast solution (400 mg/kg of body weight, subcutaneously), the temperature decrease induced by immersing the animal in cold water (4oC) or the introduction of reserpine (5 mg/kg, intraperitoneally). Investigation of the effect of peptides with PEFC. ID. N 2, 3, 4, 6 (10 ng/kg body weight, intraperitoneally) in their introduction to high and low temperature showed that they normalize both temperature violations.

14) Impact on krovetvornoe.

White mice by the injection was administered cytostatic funds (Endoxan, vincristin, adriablastin, cytarabine) in doses LD50(dose, affecting half of the experimental animals). One hour before administration of cytostatic means the mice were injected peptides with PEFC. ID. N 2, 3, 4, 6 in the amount of 10 μg, whereas the control group of mice received the same volume of saline. Animals were killed on the 3rd, 5th, 7th and 11th day after the experiment. Apologia liver and spleen.

On the third day of the experiment, differences between groups of animals were not observed in the values of L, E, Tb and neutrophils. On the contrary, healthy blood-forming cells in the bone marrow of animals who introduced the peptides strongly contrasted with aplasia observed in the control group of animals.

The number of neutrophils and leukocytes was significantly increased in animals that were injected peptides, up to 5-th day, reaching normal values on the seventh day, whereas the control group did not observe the normalization until the 11th day.

15) Impact on reducing pain.

Pain was induced by intraperitoneal administration (0.2 ml each mouse) 2%-aqueous solution of MgSO4or 0.6% acetic acid. Introduction peptides with PEFC. ID. N 2, 3, 4, 6 (10 μg or 10 ng/kg, intraperitoneally) on different circuits simultaneously with the induction of pain or after 30 min resulted in a significant decrease in the duration of pain compared with the control groups of animals.

16) Effect on infection with rickettsiosis.

0.2 ml of the suspension Coxielle Burnetti (dissolved in 10-9 times the volume) was administered by injection into oocytes. In these oocytes were injected peptides with PEFC. ID. N 2, 3, 4, 6 (1 PG, 10 PG, 100 PG and 1 ng). Survival time was significantly Bolshoi melanoma b-16) were cultured in vitro under standard conditions. The effect of peptides with PEFC. ID. N 2, 3, 4, 6 on the growth of these cell cultures was investigated in vitro. 2 days after initiation of cultivation in culture added these peptides with a concentration of 3% (0.1 ml). After 3 days was determined by count of the number of cells. The results showed an inhibitory effect of these peptides on cells of melanoma b-16.

b) the Tumor ascites Ehrlich (EAT) is a tumor that can grow in all strains of mice, as in ascitic and solid form, depending on the route of administration of tumor cells. Studied the effect of incubation of cells EAT with peptides with PEFC. ID. N 2, 3, 4, 6 on the survival time of mice that were injected these cells subcutaneously or intraperitoneally. Animals were examined during the period of 45 days. A control group of 15 male mice (NMRI strain) received 0,h cells EAT. Before the introduction of tumor cells were incubated for 1 h at 4oC in saline. The volume of injection was 0.2 ml, the Average survival time in this group was 35 days and only 3/15 them survived 45 days.

The experimental group of 15 male mice of the NMRI strain were injected the same dose of tumor cells in the same volume, but these cells were pre-incubated in a solution of the indicated peptides (2 μg/ml). the e more than 45 days. The difference between the control and experimental groups are significant (P<0,01). The same results were observed, if you applied the same methodology and EAT were injected intramuscularly (instead subcutaneous injection).

c) To study the effect of peptides with PEFC. ID. N 2, 3, 4, 6 (injected at 10 mg/kg body weight, intraperitoneally) on lung metastases induced by injection of melanoma cells or aplastic carcinoma of the breast, used mice. In the group of mice that were injected peptides observed a significant decrease in the number of metastases.

Based on the above data it can be stated that these peptides prolonged survival time of animals with tumors having antitumor activity.

18) Antidepressant activity.

For detecting antidepressant activity was used to test for dizziness in accordance with the publication Porsolt at. al., Eur J. Pharmacol, 47, 379-391, 1978.

For this experiment we used male mice Wistar (180-240 g). The target peptide with PEFC. ID. 4 introduced by the described method. In short, control animals were brought into the normal position, the determination was carried out for 5 min and counted the time of immobility. Time nelco 60-70 C. You can calculate the following graph of sensitivity to dose: 10 μg - 10 ng/kg of body weight - full impact, 10 PG activity is still present, 1 PG/kg - exposure is not observed.

The results of pharmacological studies suggest that these peptides protect the body from stress and diseases, common normalizing organic functions.

They can also be used to prevent and treat diseases and disorders of the bodies of man and animal. More specifically, these compounds will be used for the treatment of:

stress-induced disorders, disruption and damage to the liver and pancreas atrophy of the testicles and sperm motility, impaired fertility, improvement of commercial livestock disease rickettsiosis, thyroparathyroidectomy, diabetes, disorders of the adrenal glands, disorders in the blood system, blood clotting disorders, bleeding disorders, postcastration/menopausal disorders, ischemic and toxic injury, mental disorders, hypertension, disorders of body temperature, pain, swelling, depression.

In General, describes the PE the or environment, for example, a filler, non-toxic buffer, or physiological saline solution. Such pharmaceutical preparations can be used local or systemic way. Drugs can be in any suitable form, for example in the form of liquid, solid or semi-solid form, in the form of an injection solution, tablets, ointment, lotion, capsules, in the form for sublingual injection or other similar form.

These peptides will be used in the range from 10-5up to 10-2mg/kg of body weight with the systemic administration. The local introduction of them will apply to a larger number, such as 0.1 to 0.5%.

Very favorable for the application of the studied peptides is the absence of any symptoms of toxicity when introducing them to 50 mg/kg of body weight, and the high activity of these compounds in oral (intragastric) administration.

Described peptides can be synthesized stepwise condensation of protected amino acids in homogeneous liquid system or, preferably, in the solid phase. For the synthesis of cyclic peptides receive first semi-protected linear peptide of desired length in the form Olkiluoto ether at the C-terminal part of the molecule, which in turn azide. Azide then nd the peptide with the free terminal groups cyclist using diphenylphosphinite in very dilute solution.

Further, the invention is illustrated by way of examples, however, the scope of the present invention is not limited to the given examples.

Example 1. Synthesis of a peptide with the application of the protective radical Boc.

The peptide synthesis was performed using 100 IGY Boc-Val-(PAM) polyacrylamide resin to obtain a peptide with C-terminal-carboxypropyl (PAM purchased from Applied Biosistems, replacing it is 0.56 mEq/g aminotriazoles, substituted aminoacyl-4-(oxymethyl)phenyloxazol acid). Boc-amino acids (Boc-tert-butoxycarbonyl) are condensed sequentially on the polymer carrier using diisopropylcarbodiimide as a condensing means. In each stage Boc was removed by treatment of the product 50% solution triperoxonane acid (TFA) in dichloromethane. The amino group was then deprotonirovaniem using diisopropylethylamine.

The transformation in each stage should be higher than 99.5% pure. If the transformation was not enough, it was repeated. After the complete synthesis of the peptide was tsalala from media way with low content of HF (2 hours at 0oC). As acceptor ion Carbonia used anisole. HF was evaporated by blowing nitrogen. For separation of the crude peptide oily residue is poured in with the th using column size h mm, filled with silica gel RP-18. For gradient elution using solvent system: a 0.1% solution of TFA in water/acetonitrile. Detection was performed by UV absorption at 225 nm. Synthesis of a peptide with PEFC. N 4 shown in Fig. 1.

Example 2. Synthesis of a peptide with the application of the protective radical Fmoc.

For the synthesis used conventional Fmoc-protected amino acid (Fmoc-9-fluorenylmethoxycarbonyl). Side chains were protected by conversion to butyl esters (Asp, Apm, Glu, Aad) and Boc-derivative (Lys). The first amino acid residue (Val) combined with the polymer carrier, which was used BHA-resin (BHA-resin-benzhydrylamine) using diisopropylcarbodiimide as a condensing means. At each stage of the protective Fmoc group was removed with piperidine. Then the second and all following amino acid residues were introduced in the same way until completion of the total synthesis. For cleavage of the peptide used a mixture of TFA/TFMSA/anisole in the ratio 2:17:52 (vol/vol).

The peptide was then purified by the method GHUR described in example 1. Synthesis of a peptide with PEFC. ID. N 2 is illustrated in Fig. 2.

Example 3. Synthesis of a peptide with the application of the protective radical.

All amino acids used would be the and the radical Z (Z-benzyloxycarbonyl) for lysine and tert-bootrom (tert-butyl ether) for aspartic and glutamic acids. Media Morrifield (sewn klimatisierung gel polystyrene) with a capacity of 1.4 mmol/g was used for linking the first Ddz-acid through its cesium salt. After condensation with the use of dicyclohexylcarbodiimide (DCC) protective radical Ddz removed at each stage of a 5% solution of TFA in dichloromethane. The product is then washed and deprotonirovaniem 10% solution of triethylamine in dichloromethane. After deprotonation spent the next stage of condensation in the same way. These stages of condensation was repeated to obtain the target sequence.

At the end of the synthesis the peptide was tsalala from the polymer carrier by using a mixture of HBr /TFA/anisole. After evaporation of the volatile part derived peptide besieged from dry diethyl ether and dried. The peptide was then purified by the method GHUR described in example 1.

Synthesis of a peptide with PEFC. ID. N 6 illustrated in Fig. 3.

Example 4. The synthesis of the cyclic peptide.

Partially protected peptide of the formula

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previously received by the method described in example 1 or 2.

0,0005 molar solution of this compound in dimethylformamide was cyclically after adding diphenylphosphinite and triethylamine at 25oC for 12 h

The solvent is carefully removed and the product was purified by way GHUR described in example 1.

Cyclic peptide with PEFC. N 10 formula

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was obtained with the yield of 10%.

Synthesis of linear and cyclic peptides using the protective radical.

In the same way using protective groups Ddz synthesized linear and cyclic peptides. Side groups defended the radical Z (a lysine residue) and benzyl ether group (the remainder of glutamic and aspartic acids). As the polymer carrier used resin HYCPAMR(trademark ORPEGEN, Heidelberg, Germany), which is 4-bromocrotonic- -alaninaminotransferaza. The first amino acid (Ddz-valine) attached to the polymeric carrier through its cesium salt.

Then the radical Ddz removed using triperoxonane acid (5%) in dichloromethane, the amino group was deprotonirovaniem diisopropylethylamine. At the next stage, Ddz-glycine was coupled with a valine residue in the polymer matrix using diisopropylcarbodiimide (DIPC) in the presence of 1-hydroxybenzotriazole.

These steps were repeated to obtain the target peptide. Synthesized peptide in protected form then gently is the lack of oxygen. Additionally similarly morpholine was used as acceptor for allyl groups.

Polymeric media separated by filtration and washed with tetrahydrofuran. The solution is then filtered through a short column with silica gel to remove the palladium catalyst.

The eluate, which contains the partially protected peptide with PEFC. 4A, dried after evaporation in vacuo of the solvent.

Synthesis of the linear peptide with the sequence 1C.

Partially protected peptide with the sequence 4A was dissolved in 2,2,2-triptoreline and was first made in the presence of palladium on coal (10%) hydrogen at 30oC. the Catalyst was removed by filtration, the solvent was evaporated in vacuum. The residue is then purified by the method GHUR as described in example 1. After cleaning got peptide with PEFC. ID. # 4. The product was identical with the compound obtained in example 1.

The synthesis of cyclic peptide with PEFC. N 9.

Partially protected peptide with PEFC. 4A was dissolved in a mixture of dimethylformamide and dichloromethane (1:) for the formation of 0.001-molar solution. For cyclization were added DIPC and butyl alcohol and the mixture is left to stand at 20oC for 10 h Then the solution was concentrated until the om LH-20. The fractions containing partially protected cyclic peptide with PEFC. 9a, was selected and was first made using palladium on coal (10%) as catalyst at 30oC passing through the system of hydrogen with vigorous shaking for 8 h

Then the catalyst was removed by filtration and the filtrate was dried by evaporation of the solvent in vacuo. The obtained peptide was further purified by way GHUR described in example 1. Pure cyclic peptide identified as the peptide with PEFC. N 9.

The synthesis of the polymer carrier HYCRAM was performed using a protective radical Ddz. The cyclization reaction is illustrated in Fig. 4 and 5. All synthesized peptides tested for purity by way GHUR using columns with silica gel RP-18 (octadecyltrimethylammonium). As eluent used a mixture of solvents containing water, acetonitrile, triperoxonane acid. Used to do conventional gradient elution. In all cases, the purity was above 95%.

The peptides were characterized by amino acid analysis (these values differed from theoretical 10%), analysis on amino acid sequence, molecular weight, which was determined by FAB-mass-spectrometrie specific details, it should be clear to everyone who works in this field that you can put into practice many other examples of the invention and many changes to make. The essence and scope of the invention are presented in the claims.

Example 5. Peptides with the sequence N 1, 3 and 5 are very similar, and therefore, they are synthesized in accordance with the same methodology that accurately described in example 2, but using for each peptide 1, 3 and 5 in the corresponding amino acid sequence describes how to add a protected amino acids. These unprocessed peptides are then purified using the method Ehud described in example 1.

UV and IR spectra of the synthesized peptides with the sequence N 1, 3 and 5 shown in Fig. 8, 9 and 10.

Example 6. Peptides with the sequence N 7, 8 and 11 are shorter and similar peptide chain and synthesized in accordance with the procedure described in example 1, but use for each peptide 7, 8 and 11 in the corresponding amino acid sequence describes how to add a protected BOP amino acids. These peptides are then purified using the method Ehud described in example 1, UV and IR spectra of the synthesized peptides with posledovateley necessary UV - and IR-spectra of peptides with a sequence of N 2, 9 and 10 in Fig. 14-16.

To confirm p. 12 relating to pharmaceutical compositions, the applicant is also required examples.

Example 7. Manufacture of capsules. Capsules containing synthetic peptides with the sequence N (numbers) of the present invention, manufactured in accordance with methods already known in pharmaceutical technology.

The following describes the composition of the capsules, which is used as an example, peptide 2, but instead, you can use any of the peptides I - II:

Peptide with sequence N 2 - 0.05 mg

Lactose - 99,0 mg

Magnesium stearate 0.5 mg

Total - 100 mg

or:

Peptide with sequence N 2 - 0.3 mg

Peptide with sequence N 9 - 0.3 mg

Lactose - 99,0 mg

Magnesium stearate 0.4 mg

Total - 100 mg

In accordance with this method, each peptide is synthesized using the present invention can be processed into capsules.

Example 8.

The manufacture of tablets.

Pills containing synthetic peptides of the present invention with a sequence of N 1-11, manufactured in accordance with conventional methods already known in BR> Lactose - 37,5

Starch - 6,0

Talc - 3,0

Tragacanth - 2,5

Magnesium stearate and 0.5

Total - 50

Example 9. Manufacturing cream

Cream containing a therapeutically active amount of peptides with a sequence of N 1-11, manufactured in accordance with conventional methods in the pharmaceutical industry.

The composition of the cream next, g:

The peptide of the present invention is 2,5

Emulsifier - 25,00

Hydrogenated peanut oil - 60,57

tween 60 - 131,60

Propylene glycol - 30,00

Methylhydroxybenzoate - 0,16

Propylhydroxybenzoate - 0,17

Total - 250

Example 10. Manufacturing solution for injection

Solution for injection containing a therapeutically active amount of a synthetic peptide with the sequence N 1-11 of the present invention receive in accordance with the usual technique already known in the pharmaceutical industry.

The composition of the solution is mg:

The peptide of the present invention is 0.3 to

Sodium salt of carboxymethylcellulose - 1,5

Polyvinylpyrrolidone - 5,8

Lecithin - 2,7

Benzyl alcohol - 0,1

Buffer - as you need

Double-distilled water to a final volume of 1.0 funds for insertion

The composition is a solid tool for the introduction, mg:

Peptide with sequence N 2 - 0,3

Peptide with sequence N 9 - 0,3

Mannitol - 10,0

Total - 10,6

The crude compound was dissolved in 1 ml of double-distilled water, sterile filtered in bubbles and lyophilizers in a vacuum.

In some cases it may be advantageous to prepare a combination of two peptides with a similar activity but with different duration of therapeutic effect, in order to obtain a therapeutic form of long and homogeneous activity.

The peptides of the present invention was studied on various therapeutic models, using intraperitoneal, intravenous, oral and local application method. Usually for the initial studies used a sterile filtered solution or cream.

1. The peptides of General formula I

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containing from 8 to 15 amino acid residues in the chain,

where Xaa is residue of neutral aliphatic amino acids like Ala, -Ala, Leu, Ile, Gly, Val, Nle, Nva;

Yaa - amino acid residue of the basic character, such Lys, Arg, Orn, His;

Zaa - residue amino acid character, such Glu, Asp, Aad, Apm,

3. The peptide according to PP.1 and 2, having the sequence in circuit with identifying number 1

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4. The peptide according to PP.1 and 2, having the sequence in circuit with an identifying number 2

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5. The peptide according to PP.1 and 2, having the sequence in circuit with an identifying number 3

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6. The peptide according to PP.1 and 2, having the sequence in circuit with an identifying number 4

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7. The peptide according to PP.1 and 2, having the sequence in circuit with an identifying number 5

< / BR>
8. The peptide according to PP.1 and 2, having the sequence in circuit with an identifying number 6

< / BR>
9. The peptide according to PP.1 and 2, having the sequence in circuit with an identifying number 7

< / BR>
10. The peptide according to PP.1 and 2, having the sequence in circuit with an identifying number 8

< / BR>
11. The peptide according to PP.1 and 2, having the sequence in circuit with identification number 9

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12. The peptide according to PP.1 and 2, having the sequence in circuit with an identifying number 10

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13 Peptide on PP.1 and 2, having the sequence in circuit with an identifying number 11

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14. Peptides under item 1 of the formula I where at least one and at most seven amino acid residues 1 through 15 are missing.

15. Peptides under item 1, obladaushi the m action based on the peptide and the usual additives, characterized in that the peptide contains one or more compounds according to paragraphs.1 - 15 in effective amounts.

 

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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 medicine, molecular biology and Virology and represents the biologically active compound is a synthetic oligopeptides corresponding to amino acid residues (and

The invention relates to new biologically active compounds, specifically to a peptide of General formula: R-GLu-Leu-Lys-Pro-Leu-X-Glu-Val-Leu-Asn-Leu-R', where R is CH3CO-(Ac), Ac-Glu, Ac-Leu-Glu-Glu-, HCO-(Form), Form-Glu-, Form-Glu-Glu, Form-Leu-Glu-Glu-, CH(CH)nCO-CH(CH)nCO-Glu-, CH(CH)nCO-Glu-Glu-, CH(CH)nCO-Leu-Glu-Glu-, n - 8 - 18, X - L-Glu, D-Glu, R' - OMe or NH2who are the inductors and/or potentiators growth stimulating activity of macrophages in vitro, have regenerative-reparative action in vivo, and can find application in medicine for treatment of several diseases involving generalized activation of macrophages (hepatitis, toxic liver damage, cirrhosis of the liver, skin wounds, burns, polytrauma, diabetes and others)

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

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

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

TrpAlaGlyGlyAspAlaSerGlyGlu

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

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

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

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

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

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

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

AA2leucine, arginine, ornithine, or glutamic acid;

AA3aspartic acid, N-metilparabena acid;

AA4isoleucine, phenylalanine;

AA5isoleucine, N-methylisoleucine;

AA6tryptophan, N-formylthiophene

or their pharmaceutically acceptable salts

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

The invention relates to medicine, namely to funds on the basis of biologically active substances of plant origin, and can be used as a means of providing anti-inflammatory, immunomodulatory, anabolic effects
The invention relates to medicine, namely the subject of the invention are tablets based on partially or fully water-soluble natural and/or synthetic polymers selected from rubbers, alginates, Cartagena, starch, pectin and gelatin containing poly-(dimethyl-siloxanes) (Dimethicone, simethicone), and the method of their manufacture

The invention relates to the protection of certain 3-substituted-2-oxindole-1-carboxamide formula

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and their pharmaceutically acceptable basic salts, where X is H, Cl, or F; Y is H or Cl; and K represents benzyl or thienyl, each optionally substituted by Cl or F; from decomposition in the presence of light using a dye absorbing light or packing material

The invention relates to pharmaceutical industry
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