Analogues of lhrh antagonists and methods for their production


(57) Abstract:

The inventive new LHRH antagonists of the formula [Nc - D2Nal1- DpClphe2- D3pal3- Ser4- Mop5- D3pal6- Leu7- Ary8- pro9- Dla10]NH2or [Nc - D2Nal1- Dphe2- D3pal3- ser4- Mop5- D3pal3- Leu7- Arg8- pro9- Dla10] NH2or [Nac - D2Nal1- DpClphe2- D3pal3- Ser4- Arg5- D3pal6- Leu7- pap8- pro9- Dla10]NH2. New LHRH antagonists of the formula [Nc - D2Nal1- AA2- AA3- Ser4- AA5- AA6- Leu7- AA8- pro9- Dla10] NH2where DNal : DArAla, where AA2- DArAl, where AA3: DArAla, where, or where AA5Is Arg, Tyr, or DArAla, where, or where,- N(R1R2), R1and R2- CH3CH3CH2C3H7C4H9; AA6- Drphe, where, or where, or NR1R2where R1and R2- CH3CH3CH2, AA8- Arg or DArAla, where, or where, or NR1R2where R1, R2- CH3CH3CH2C3H7C4H9as new peptides can be used in the treatment of diseases of the endocrine sisb receiving LHRH antagonists , namely, that C-terminal N-protected amino acid attached to benzhydrylamine resin, otscheplaut protective group and continue to build up the peptide chain in the sequence corresponding to the peptide of formula IV under conditions of solid-phase synthesis, and from the resulting peptidylarginine otscheplaut peptide. Get peptides with valuable properties, with activity in the stimulation of histamine reduced to such a level to meet clinical requirements. 3 S. p. f-crystals, 6 ill., table 4.

The present invention relates to new peptides and their derivatives having the exact chemical structure. The invention also addresses the methods of their production and application. Hypothalamic hormone that stimulates the secretion of luteinizing hormone (LHRH), acts on the anterior pituitary, stimulating the secretion of luteinizing hormone (LH) and follicle-stimulating hormone (FSH). Antagonistic analog of LHRH acts on the anterior pituitary quickly, lasting, can be safely and reversibly used for contraception or selective suppression of the secretion of gonadotropic hormones. In this application, LHRH antagonists dominate agonists. To date, the predicted ioctl in relation to the stimulation of histamine (HRA). He called passing swelling of the face and limbs in rats when applied in doses of 50 to 100 times therapeutic dose. The results of clinical trials have demonstrated the systemic effects of histamine. Other antagonists GSLG containing DArg6or DLys6showed similar side effects, ED50for HRA were below 1 µg/ml. the Present invention represents a new antagonists, which have a very high antiovulatory activity (AOA) and very low activity in relation to the stimulation of histamine HRA and minor side effects. The content and examples of this invention include the following.

The methodology of the forecast structure of this invention is based on the topological similarity of the molecules of the starting compound (NAc - D2Nal', DpClPhe2, D3Pal3Ser4, Tyr5, DArg6, Leu7, Arg8Pro9, DAla10)NH2(II) and the neuropeptide substance P, which is a characteristic black modifications such as alkaline and lipophilic region of the molecule starting compound to obtain a new antagonists with as high AOA and low GAW. The term modification means a building or replacement minocin structure consists in the introduction of a suitable alkaline group and substitution of natural unusual amino acids in position 2, 3, 5, 6, 8 in (II).

The following is also included in the methods and examples of this invention.

1. The substitution of D3Pal, which is an aromatic amino acid having a suitable alkalinity, instead DArg8in (II) to obtain the analogue of (III): (NAc-2DNal', DpClPhe2, D3Pal3Ser4, Tyr5, D3Pal6Ser7, Arg8Pro9, DAla10)NH2< / BR>
2. Substitution of Tyr5for Arg5in (III) for (IV): (NAc-D2Nal', DpClPhe2, D3Pal3Ser4, Arg5, D3Pal6, Leu7, Arg8Pro9, DAla10)NH2< / BR>
3. Substitution D3Pal3in (IV) on DPhe3or its derivatives DXCH2Phe to receive (V): (NAc-D2Nal', DpClPhe2, DPhe3Ser4, Arg5, D3Pal6, Leu7, Arg8Pro9, DAla10)NH2or its analogues (DXCH2Phe3)

4. Replacement DPhe3and its derivatives D3Pal3in (III) to obtain (V'): (NAc-D2Nal', DpClPhe2, DPhe3Ser4, Tyr5, D3Pal6, Leu7, Arg8Pro9, DAla10)NH2or its analogues (DXCH2Phe3)

Synthesized a series of new antagonists of LHRH with formula (NAc-D2Nal', AA2, AA3Ser4, AA5, AA6. Leu7, AA8, DAla10)NH2where AA are at the end of the text.

The LHRH antagonists obtained using the above method, as peptide drugs, can be used in the treatment of diseases of the reproductive endocrine system, such as endometriosis, premature puberty in children, and for the treatment of prostate cancer and breast cancer, as well as as male and female contraceptives for birth control, or used for the diagnosis and treatment of infertility, etc. Such peptide drug can be prepared in the form of conventional injectable, injectable, capsules or other dosage forms for practical use.

Further description of this invention is as follows.

When the natural path of excretion of histamine in the body is very important neuropeptide substance II, its ED50for HRA equal 5-15 μm. The chemical structure of SP is a (Arg', Pro2, Lys3Pro4, Gln5, Gln6, Phe7, Phe8, Gly9, Leu10, Met11)NH2.

Study of the relationship between its structure and HRA showed that Arg1-Pro2-Lys3on the N-end of the molecule SP significantly to the HRA, as removing these trlat C-end of HRA remained at one-fourth from the initial HRA. In addition, deletion of Phe8and Phe7reduces HRA 4% and 0.5% of such JV. Next remove Gln5,6does not cause significant changes to the HRA. The above data indicate that the lipophilic region around Phe7,8determines the level of the HRA, this area is involved in the binding of the molecule to the receptor of mast cells.

As mentioned earlier, the analogues of (D2Nal', DArg6)LHRH showed very high HPA, their molecular structure is topological similarity to SP: DArg6- Leu7- Arg8in the first, seems to be consistent with Arg'- Pro2- Lys3in the past, both consist of a pair of strongly alkaline amino acid residues, among which only one neutral amino acid residue, i.e., both analog LHPH - 2DNal', DArg and SP contain two strongly alkaline amino acid residue, which are arranged one against another in positions 1,3. On the other hand, the dispersion of aromatic amino acid residues in the first, say, corresponds to the region of Phe7,8in SP, from the point of view of determining the value of the HRA.

The structure of this invention consists of two aspects: one is the modification of the site Tyr5- DArg6- Arg8C-end, and the other is a fine selection of aromatic kid6, Leu7, Arg8Pro9, DAla10)NH2(II) used as starting material, which showed AOA 100% at the dose of 0.5 μg in corn oil, 5% at 0.25 μg.

First of all, DArg6in (II) could be replaced by a weak basic or neutral aromatic acids such as D3Pal, D6Qal tetrahydrolipstatin, methyltryptophan. (NAc - D2Nal', DpClPhe2, D3Pal3,6, DAla10)LHRH (III) was obtained when D3Arg6in (II) was substituted for D3Pal6. (III) showed AOA 100% at the dose of 3 mg, 83% at 1 μg (in corn oil), and ED50by the HRA was equal to 9.8 µg/ml, which is much better than analog "Nal-Arg" ED50by the HRA was less than 1 µg/ml apparently, in order to obtain a high AOA, the alkalinity of the whole molecule must be equal to or close to one of the pair of arginine. Since the position 5 and position 6, is not involved in binding with the receptor, in position 5 may be placed in a wide range of amino acids, including arginine. Were created a series of new analogues. For example, if the substitution of Tyr5for Arg5in (III) is obtained (NAc - DNal', DpClPhe2, D3Pal3,6, Arg5, DAla10)LHRH (IV). Both (IV) and (II) contain two arginine residue, but the distance between the two residues in (IV), where the geometric waymores is, probably, the HRA will be reduced and, on the other hand, due to the presence of two residues AOA should not be lower than (II). The results of biological studies (IV) showed that the ED50by the HRA was equal to 3.5 g/ml, whereas AOA was 60% at 0.12 µg (corn oil), 85% at 0.25 μg, 100% at 0.5 mg. This was the first opportunity for LHRH antagonists, when achieved ED50for AOA was equal to or less than 0.125 mg.

Therefore, further creating patterns based on the structure (IV).

In the molecule (IV) there are four residue with alkaline ions, D3Pal3,6and Arg5,8whereas (II) contains only three alkaline balance. Therefore, it is acceptable to replace one D3Pal neutral amino acid; on the other hand, (IV) showed a very high hydrophilicity, and the decrease in hydrophilicity substitution DPal (IV) a hydrophobic amino acid, probably, will be beneficial to delay the drug in the body and increase the duration of the effect. Then created a new series of analogues by replacing instead of DBPal3. (V) showed 100% AOA at the dose of 1 μg (in saline solution) equal to activity of the parent substance (IV), whereas the HRA is reduced by half: ED50by the HRA was to 7.4 µg/ml.

Additional samewe - Leu7- Arg8C-end (IV) apparently play a major role in the starting mechanism of secretion of histamine. D3Pal combines in one molecule of the aromatic properties of the structure, alkalinity and hydrophilic nature, he also Staropramen in LHRH antagonists for the receptor binding. Thus, the creation of a new series of unnatural amino acids using such properties as (V) D3Pal, can lead to better antagonists than (IV) or (V).

Modification of natural lipophilic aromatic amino acids such as phenylalanine, for example, by the method described below in the section Synthesis of unnatural amino acids, leads to the creation of a series of new amino acids, which combine properties of aromatic compounds, hydrophilic and alkaline properties in a single molecule and can be expressed by the formula


(VI), where

R1and R2may be the same or different, can be represented by a chain or a loop may also contain a heteroatom. When you change R1and R2can be obtained a series of amino acids, which are constantly systematically changed alkalinity, hydrophilicity and stereoscreen. The introduction of these aminocyclo studies have shown, each series has given at least one new antagonist, showing 100% AOA at the dose of 1 μg, like (IV), at the same time was significantly reduced. An example was (VII): (NAc - D2Nal', DpClPhe2, D3Pal3Ser4, Mop5, D3Pal6, Leu7, Arg8Pro9, DAla10)NH2,

which showed 100% AOA at 1 µg, ED50when GSA 14,7 mg/ml, is clearly better than (V). When replacing Arg8in (IV) to (VI) reduction HRA positively correlated with length P in (VI), so that the ED50on the HRA could be higher than 200 μg/ml, such compounds can easily be dissolved in an aqueous solution and, as expected, will be used in the clinic without complications dosage forms. The results showed that Arg8or Lys8not significant for LHRH antagonists with high potency. The corresponding alkaline center in position 8 can provide a high AOA, meanwhile, activity of inducing the production of histamine mast cells was significantly decreased when was built the group with the basic properties mentioned above, creating a significant stereoremaster.

This invention, by combining the modification on the N end, and at the end of C, leads to the creation of the best antagonists of LHRH.

The process of synthesis of D - or L-amino acids generated and synthesized in four ways by synthesis, presented in the diagram below (see diagram B in the end of the text). The structure of these unnatural amino acids are shown as the main structural formula listed in the same scheme. Some of these amino acids possess alkalinity, hydrophilic and aromatic properties, respectively, whereas others combine them all in a single molecule.

2. Synthesis of peptide

The synthesis begins with the C-Terminus of the peptide on benzhydrylamine-cleaners containing hydrochloride resin (BHA-resin) using solid-phase peptide synthesis, the proposed Merrifield. This three-phase process involving attachment, interaction and dissociation. Dichloromethane (DCM) is the main solvent used for washing before each next stage of the reaction, at the same time, the alcohol is isopropanol (IPA) and N, N-dimethylformamide (DMF) were also used where necessary. The binding reaction is carried out with catalysis by excess dicyclohexylcarbodiimide (DCC) adding a sufficient amount of 1-hydroxybenzotriazole (HOBT). The degree of binding assays was determined by ninhydrin method Cases. The second binding reaction should be carried out, if it gives a positive result in the sample Cases. The peptide chain was useplease from smo is rebaudi binding resin, at the same time removes all temporary protecting group. After washing with ethyl acetate or ether crude products LHRH-antagonists obtained by extraction with an aqueous solution of acetic acid followed by lyophilization. Output in excess of 50%.

3. Purification of peptide

(1) the Peptide is purified using gel chromatography or distribution chromatography on a silica column high 60 - 100 cm during the inspection using a UV/TLC. Cleaning LHRH-antagonists was performed once and the product was obtained after lyophilization of the major fraction. The output is 50 - 90% and the purity can be over 90%.

(2) the Peptide was then purified on the device for high-performance liquid chromatography (HPLC) waters, using a column address phase C18 (of 7.8 x 300 mm) (m Bondapak 84176). The output at this stage is 20 to 50%, with a purity not less than 99%.

4. Analysis of the purity of peptide

(1) Analysis TCX

It is held on a plastic plate coated with silica gel 60F254, length 5 - 10 cm In all cases revealed a single spot when working with five different solvent system.

(2) before HPLC Analysis

In all cases, detected a single peak at the elution of the two types of solvent system, and ( = 210). The size of the sample was 10 - 200 mcg.

5. Amino acid analysis of peptide

In accordance with the method of PICO-TAG developed by waters, 50 µg of the sample, which was dried under vacuum for 2 hours and accurately weighed on the pan balance with a resolution of 10-5, After dissolution in water of 10 ág of sample added to the reaction tube, in which, according to the guide, added hydrochloric acid 1:1 (containing 1% phenol). The reaction lasts for 20 - 24 h at 105oC. the Reaction lasts for 22 to 24 h at 105oC in tightly closed container, which is filled with nitrogen and evacuated under vacuum to remove oxygen from the reaction tubes. Isothiocyanate phenol is added to obtain the amino group after evaporation of excess amounts of hydrochloric acid. He then analyzed using a liquid chromatograph, equipped with the amino acid analytical column PICO-TAG with a UV detector ( = 254 nm). The content of each amino acid and the relative molar ratio was calculated to determine the amino acid composition of the sample based on the comparison of the component parts of each amino acid with one H-standard sample waters. The classical method with ion exchange ninhydrin proletarienne results for this method, you need ten times more sample.

6. Evaluation of biological activity

Used method antiovulatory rats Corbin. In this experiment, we used healthy adult female SD rats (weight 200 - 500g). All animals were kept at 22 - 24oC and under a regime of 14 h/10 h (light/dark). They received a standard diet and water needs. In this experiment could be used rats that have identified at least two consecutive four-day estrus cycle in the study of vaginal smears. Rats were injected peptides (LHRH antagonists) in different doses in saline solution in the middle of the stage leakage.

Rats were scored on the next day, they both ovaries with two sides explored under preprofile loupe to determine the number of Mature eggs. Rats were divided into several groups according to the obtained doses, each group consisted of about 10 animals and a control group, the animals which were administered an equal amount of saline solution consisted of 9 to 10 rats. Antiovulatory activity (AOA) is shown in the following equation:

< / BR>
7. Assessment activity by the stimulation of histamine

(1) Determination of the secretion of histamine (HRT)

In this experiment, use the and CO2the peritoneal cavity was washed with 50 ml of medium PIPAS AC containing 20 units of heparin. After subsequent centrifugation at HD for 8 min at 4oC cells are again washed and finally resuspendable to a concentration of from 8 to 24105the total number of leukocytes/ml PIPAS AC. This slurry contains about 5 to 10% of fat cells. The washed cells were used immediately after selection and pre-heated for 5 min at 37oC before 0.3 ml sample pipette was placed in polystyrene tubes containing 0.3 ml of diluted peptide. Mixtures were incubated for 15 min at 37oC and the reaction was stopped by centrifugation at HD for 15 min at 4oC. Supernatant examined on the content of histamine using a fluorometric method for the determination carried out manually after sequential extraction with n-butanol and n-heptane. The content of histamine can be installed in a standard histamine curve (see below). The process of secretion of histamine can be calculated by the following equality



E is an indication of fluorometry for the experimental sample; B - indication of fluorometry for samples containing only cells and the buffer; and C what I can be obtained by drawing the graph of the values of optical density on the testimony of fluorimetry at 350 nm/450 nm (activation/fluorescence) for a concentration series of dilutions of the solution accurately weighed quantity of histamine hydrochloride. Relative parameter p standard histamine curve can be 0,9998 and the lowest detectable concentration of histamine equal to 0.5 ng/ml

The value of the ED50for peptide can be obtained from the response curves dose-response by drawing the graph of the level of secretion of histamine in comparison with the concentration of peptide in semi-logarithmic scale.

All samples of peptides must be tested with fat cells from at least 3 different rats.

(2) Skin sample for anaphylactoid activity (CAT)

In this experiment, we used healthy adult rats-females (weight 250 g). Rats were injected intravenously a solution of Evans blue (1 ml of 0.05%). Immediately after that intradermally injected with a solution of peptide (5, 0.5 and 0.05 mg/ml, respectively) or saline (control) in shaved sector on the back of animals. After 30 min after injection, rats were killed and the skin of the back straight. The diameter of the lesions was measured in millimeters in two perpendicular directions using a compass with a Vernier. The diameter control is usually less than 5,5 mm

The amount of Evans blue, penetrating into the skin of the blood vessels, may be also determined spectrophotometrically. The skin corresponding to the area the next day, the content of Evans blue in the supernatant was determined on a spectrophotometer (UV-260) at 610 nm with the reference solution acetone/saline solution (7:3). Each peptide was tested at least 3 different rats.

Using the method described above, created and synthesized a variety of new antagonists of LRRH. Briefly, the antagonists with the new structure were obtained by single or multiple substitutions of different natural and unnatural amino acids listed in the previous sections.

Some examples of new antagonists of LHRH, thus obtained, are shown in table 1.

The application of this invention

1. After completing preclinical pharmacological and Toxicological study we will be able to apply these new antagonists, which have a high therapeutic effectiveness and mild side effects, in the clinic to develop a new drug for the treatment of endometriosis and other disorders in the reproductive endocrine system, including premature puberty in children, prostate cancer and breast cancer. Because they suppress the secretion of gonadotropin by competition with endogenous LHRH binding to the receptor and act fast, reversible and safe, they may also be developed as a new type of contraceptives for men and women. Moreover, they can also be gonadotropin.

As a kind of peptide drugs, it is unlikely that antagonists of LHRH will be applied orally. But they can be easily obtained in the form of dried powder, easily soluble in saline solution for injection intravenously, subcutaneously or intramuscularly.

In addition, we study long-acting delivery system of medicinal substances, such as biodegradable, injectable capsules. Capsules can be implanted subcutaneously using a special syringe and must be absorbed by the tissue after extraction of the total number of peptide and do not require surgical removal. Long-acting delivery system is particularly useful for long-term use of LHRH analogues in the clinic.

The following is an analysis of the results from examples (using three analog IV, V, VII as typical examples).

(1) Clean

Thin layer chromatography (TCX)

On each of the chromatograms obtained with the five different solvent system, was attended by only one spot.

High-performance liquid chromatography (HPLC):

On each of the chromatograms at two different elution solvent system is present only the levels of Fig. 1 to 4.

Solution A + 80% acetonitrile

The solvent system 2

Solution A is 0.01 M KH2PO4an aqueous solution (pH 3)

Solution B - 20% solution A + 80% acetonitrile

System solutions for TCX

1. n BuOH/EtOA/SPLA/H2(5:5:1:1)

2. n BuOH/n-BuOH/SPLA/H2O(2:8:2:3)

3. n-BuOH/HOAc/H2O (4:1:5), the rising phase

4. n-BuOH/HOAc/H2O (4:1:2)

5. n-BuOH/EtOH/HOAc/H2O(1:1:1:1)

(2) Amino acid analysis

The analysis was carried out according to the method of classical'IEN and the new PICO-TAG, the results are presented in table 3 and Fig. 5, 6.

(3) the Results of biological studies

The results of biological studies, including antiovulatory activity at different doses and ED50activity the stimulation of histamine in vitro is shown in table 4, which lists 26 antagonists as examples.

As shown and described above, antagonists GSLG created and synthesized in accordance with this invention find very good properties. They are clean on the results of the analysis of TLC or HPLC. Their structure is correct, i.e. it is the same as planned. Their contraceptive activity is high: they can inhibit ovulation in rats by subcutaneous injection in the pose from 0.1 to 2.0 microns is about the activity of the stimulation of histamine in vitro (effective dose for the fat cells of rats stimulate secretion of 50% histamine) is in the range of 5 to 300 μg/ml; defeat, resulting in cutaneous anaphylactoid test in rats is low enough that meets the clinical requirements. They are very well soluble in water, all bosslaguna conducted with solutions of drugs in saline solution, so they are easy to get in injectable dosage forms for clinical use. They are also easy to prepare in the form of long-acting delivery systems, among which injectable microcapsules most convenient for long-term suppression of gonadotropin and sex hormones. Therefore, they can be used as effective with retroactive effect and safe contraceptives for men and women. They can also be used for treatment of various diseases associated with disorders of the reproductive endocrine system, such as hormone-dependent prostate cancer, breast cancer, endometriosis, premature puberty in children. They are also useful in the treatment of infertility. New LHRH antagonists can also be used when the underlying physiological and pharmacological studies of the reproductive sphere, such as in the study of funk


In the text of this patent application document, we used the following abbreviations

Ala alanine

AOA antiovulatory activity

Arg arginine

Bap dibutylaminoethanol

BOC t-butyloxycarbonyl

BuOAc butyl acetate

CAT cutaneous anaphylactoid sample

DCl dicyclohexylcarbodiimide

DCM dichloromethane

D2Nal D -- (2-naphthyl)alanine

D3Pal D -- (3-pyridyl)alanine

DpClPhe p-chloro-D-phenylalanine

DpFPhe p-fluoro-D-phenylalanine

D6Qal D -- (6-chinolin)alanine

DMF N,N-dimethylformamide

EAP diethylaminoethylamine

ED50the effective dose for 50% response

EtoAc ethyl acetate

FSH follicle stimulating hormone

Glu glutamic acid

Gly glycine

His histidine

HOBT L-hydroxybenzotriazol

VAST high-resolution liquid chromatography ninhydrin derivative

HRA activity by the stimulation of histamine

HRT determining the allocation of histamine

ISN ion-exchange chromatography c

IPA isopropyl alcohol

LH luteinizing hormone

LHRH hormone that stimulates the secretion of luteinizing hormone

Leu is leucine

Lys lysine

Map dimethylaminomethylene

Met metion the

Pep dipropylenetriamine

Phe phenylalanine

Pip piperidinemethanol

Pipes piperazine-N,N'-bis(2-econsultancy acid)

Pro Proline

Rf level offset

SE standard error

Ser serine

TFA triperoxonane acid

TLC thin layer chromatography

TR is the retention time

Trp tryptophan

Tyr tyrosine

Tep of tetrahydroprotoberberine

1. Analogues of LHRH antagonists of the formula

[NAc-D2Nal1-DpClPhe2-D3Pal3-Ser4-Mop5-D3Pal6-Leu7-Arg8-Pro9-DAla10] -NH2I


[NAc-D2Nal1-Dphe2-D3Pal3-Ser4-Mop5-D3Pal3-Leu7-Arg8-Pro9-DAla10] NH2II,


[NAc-D2Nal1-DpClphe2-D3Pal3-Ser4-Arg5-D3Pal6-Leu7-Pap8-Pro9-DAla10] NH2III

2. Analogues of LHRH antagonists of the formula IV

[NAc-D2Nal1-AA2-AA3-Ser4-AA5-AA6-Leu7-AA8-Pro9-DAla10]NH2, DNal group of the formula


< / BR>
AA2a group of the formula


in which Ar is a group

AA3group of the formula


where Ar

or a group of the formula

< / BR>

where R1, R2the same and mean CH3CH3CH2C3H7C4H9;

AA6group of the formula


where Ar

or a group of the formula

< / BR>
where X-

or group


where R1, R2the same and mean CH3CH3CH2;

AA8- Arg or a group of the formula


where Ar

or a group of the formula

< / BR>
where X-

or a group of the formula


where R1, R2the same and mean CH3CH3CH2C3H7C4H9.

3. The method of obtaining analogues of LHRH antagonists of General formula IV under item 2, wherein the C-terminal N-protected amino acid attached to benzhydrylamine resin, otscheplaut protective group and continue to build up the peptide chain in the sequence corresponding to the peptide of formula IV under conditions of solid-phase synthesis, and from the resulting peptidylarginine otscheplaut peptide.


Same patents:

The invention relates to medicine

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

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

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

sleep" target="_blank">

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


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

FIELD: bioorganic chemistry.

SUBSTANCE: invention provides somatostatin agonists of general formula: A1-cyclo{Cys-A2-D-Trp-A3-A4-Cys}-A5-Y1 (I), wherein A1 represents aromatic D- or L-α-amino acid selected from Phe, D-Phe, Tyr, D-Tyr, β-Nal, D-β-Nal, Cha, or D-Cha; A2 aromatic α-amino acid selected from Phe, Tyr, β-Nal, and Cha; A3 Lys or Orn; A4 β-hydroxyvaline, Ser, hSer, or Thr; A5 β-hydroxyvaline, Ser, hSer, or Thr; and Y1 represents NH2; amide nitrogen atoms of peptide groups and amine group of A1 in compound I are optionally substituted by methyl group, provided that at least one methyl group is available and that compound I cannot have following formula: D-Phe-cyclo{Cys-Phe-D-Trp-Lys-(N-Me-Thr)-Cys}-Thr-NH-2. pharmaceutically acceptable salts of compound I are also claimed.

EFFECT: expanded synthetic possibilities in peptide synthesis.

24 cl, 2 tbl, 18 ex

Oligopeptides // 2260597

FIELD: organic chemistry, peptides, biochemistry.

SUBSTANCE: invention describes oligopeptide or its salt taken among the group consisting of oligopeptide (1) and (2): Lys-Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu (I), Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu (II); Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp (III); Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln (IV); Ser-Ile-Glu-Gln-Ser-Cys-Asp (V); Ser-Ile-Glu-Gln-Ser-Cys (VI); Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu (VII); Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu (VIII); Gln-Ser-Cys-Asp-Gln-Asp-Glu (IX); 2) oligopeptide with amino acid sequence obtained by deleting by C- or N-end of one or some amino acids in any amino acid sequence (I)-(IX), and the modified oligopeptide representing oligopeptide biotinylated or dimerized by sulfhydryl group of cysteine residue based on oligopeptide determined in (1) or (2). Oligopeptides elicit activity with respect to hair growth stimulation.

EFFECT: valuable properties of oligopeptides.

11 cl, 6 dwg, 4 ex

FIELD: organic chemistry, peptides, medicine, pharmacy.

SUBSTANCE: invention relates to peptide derivatives named as memnopeptides that are used as an active component for manufacturing a medicinal preparation used in treatment of bacterial infection. Invention proposes compound of the formula (I): wherein radicals R1, R2, R3, R4, R5, R6, R7, R8 and (A)n have corresponding values, or its salt. Compounds of the formula (I) are prepared by culturing microorganism Memnoniella echinata FH 2272, DSM 13195 under suitable conditions in the nutrient medium containing at least one source of carbon atoms and at least one source of nitrogen atoms and the process is carrying out until the accumulation of at least one compound of the formula (I) in the nutrient medium followed by isolation of indicated compound. The attained technical result involves the development of a pharmaceutical composition eliciting an antibacterial activity. The development of the preparation provides expanding assortment of agents used in treatment of diseases said above.

EFFECT: improved preparing method, valuable medicinal properties of compounds.

10 cl, 2 tbl, 7 ex

FIELD: peptides, pharmacy.

SUBSTANCE: invention relates to low-molecular derivatives of peptides that are able to act as inhibitors in interaction between laminine and nidogen (interactions laminine/nidogen). Also, invention relates to a method for their preparing, pharmaceutical composition prepared on thereof and their using for preparing pharmaceutical agents, and for identification of inhibitors in interaction laminine/nidogen.

EFFECT: valuable properties of peptides.

5 cl, 12 dwg

FIELD: organic chemistry, amino acids.

SUBSTANCE: invention proposes agonists of somatostatin of the formula (I): X-A1-cyclo-(D-Cys-A3-A4-Lys-A6-A7)-A8-Y or its pharmaceutically acceptable salt wherein X represents hydrogen atom (H); A1 represents L-Cpa, L-Phe, L-Trp or L-Nal; A3 represents L-3-Pal or L-4-Pal; A4 represents D-Trp; A6 represents -NH-(CHR1)n-CO- wherein n = 2, 3 or 4; A7 represents L- or D-Cys; A8 represents D- or L-isomer of amino acid taken among the group consisting of Nal, Phe, Cpa and Trp; Y represents NH2; R1 represents hydrogen atom (H), and Cys in A3 is bound by disulfide bong in A wherein this disulfide bond is formed by thiol groups of each Cys residue.

EFFECT: valuable biological properties of compounds.

9 cl, 2 ex

FIELD: organic chemistry, peptides, pharmacy.

SUBSTANCE: invention relates to biologically active compounds as antagonists of somatostatin. Invention represents compound of the formula (I): A1-cyclo-{D-Cys-A2-D-Trp-A3-A4-Cys}-A-Y1 wherein A1 represents aromatic α-amino acid taken among the group: Cpa, Tfm, 1-Nal, 2-Nal and 3-Pal and substituted optionally with one or some substitutes taken among halogen atom and (C1-6)-alkyl; A2 represents aromatic amino acid taken among the group: 3-Pal and Tyr and substituted optionally with one or some substituted taken among halogen atom, (C1-6)-alkyl, OH-group; A3 represents Lys, Dab, Dap, Orn; A4 represents Thr, β-hydroxyvaline, Ser, Hser; A5 represents D- or L-aromatic α-amino acid taken among the group: Dip, 2-Nal, Trp; Y1 represents -NH2 or -MHMe wherein amine nitrogen in each amide peptide bond and amino-group A1 are substituted optionally with methyl group under condition that at least one abovementioned methyl group presents; or its pharmaceutically acceptable salt.

EFFECT: valuable biological properties of compounds.

11 cl, 1 dwg, 3 tbl, 16 ex

FIELD: chemistry of peptides, medicine.

SUBSTANCE: invention relates to preparing new peptides possessing immunomodulating, anti-proliferative, anti-tumor and antiviral activity. Invention proposes new peptides comprising up to 30 amino acid residues of the general structural formula: X1-Trp-Gly-Gln-X2 wherein X1 is taken among the following group: -His-Gly-Val-Ser-Gly-, -His-Gly-Gly-Gly-, -His-Val-Gly-Gly-, -His-Gly-Gly-Gly-Gly-, and -Gln-Gly-Gly-Gly-Gly, or absent; X2 is taken among the following group: -His-Gly-Thr-His-Gly, -Gly-Gly-Thr-His-Gly, -Pro-His-Val-Gly-Gly, -Pro-His-Gly-Gly-Gly, -Pro-His-Gly-Gly-Gly-Trp-Gly, -Gly-Gly-Gly-Thr-His-Ser, or absent.

EFFECT: valuable medicinal properties of peptides.

8 cl, 5 tbl, 5 dwg, 6 ex

FIELD: biotechnology, peptides.

SUBSTANCE: invention relates to a method for preparing antibodies raised to human leukocyte differentiation factor (HLDF) or to HLDF fragment (31-38) representing peptide of the following structure: Arg-Arg-Trp-His-Arg-Leu-Glu-Lys possessing with antigenic and nucleic acids-hydrolyzing properties, and for diagnostic aims also. Antibodies are prepared from rabbit plasma blood immunized with three injections of antigens wherein synthetic HLDF factor or conjugate is used as antigens. Diagnosis of anaplastic state of human cells is carried out by using solutions of antibodies to HLDF factor or HLDF fragment (31-38) in the concentration 0.0013 mg/ml as biological markers. Invention provides carrying out the differential diagnosis of tumors and normal organs and effective detecting initial stages in cell differentiation disturbances.

EFFECT: improved preparing method of antibody, improved method for diagnosis.

6 cl, 21 dwg, 1 tbl

FIELD: medicine, polypeptides.

SUBSTANCE: invention relates to fusion polypeptides with enhanced pharmacokinetic properties. Fusion polypeptides comprising enhancing peptide sequences associated with the core polypeptide possess with the enhanced pharmacokinetic properties, such as prolonged half-time period. Also, invention relates to methods for enhancing pharmacokinetic properties of any core polypeptide by binding the enhancer peptide sequences with the core polypeptide. Proposed core polypeptides can comprise any pharmacologically useful peptide that can be used, for example, the therapeutic or prophylactic agent. The advantage of invention involves the enhancing of pharmacokinetic properties of polypeptides.

EFFECT: enhanced pharmacokinetic properties of polypeptides.

52 cl, 18 dwg, 14 tbl, 11 ex

FIELD: medicine, oncology.

SUBSTANCE: invention relates to immunomodulator representing pentapeptide of formula Val-Val-Tyr-Pro-Asp and drug in liquid and dry forms based on the same. Both pentapeptide and pharmaceutical composition containing the same have antitumor activity in small doses and have no side effects.

EFFECT: new low molecular peptide with immunomodulating activity and antitumor preparation based on the same.

2 cl, 6 ex, 4 tbl