Oligopeptides

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

 

Technical area

This invention relates to oligopeptides with activity, promotes hair growth, and the agent, promotes hair growth, including specified Oligopeptide as the active ingredient.

Background of the invention

It was assumed that the normal morphogenesis of epithelial tissue is regulated by factors originating from the mesenchyme cells present around the epithelial tissue. Diseases resulting from abnormal morphogenesis of epithelial tissue, often caused by abnormalities of the cells of the mesenchyme. So have any interest in the elucidation of the mechanisms of regulation of morphogenesis of epithelial tissue cells of the mesenchyme. However, substances involved in the regulation of morphogenesis of epithelial tissues, showing his regulatory action depending on time and space and in the form of a complex system and therefore extremely difficult to identify these substances and to analyze their functions. It is also difficult to construct a model experimental system that simplifies the morphogenesis of epithelial tissue. For these reasons, to date there has been no substantial progress in research in this area. Thus, it is highly desirable to analyze the mechanisms that regulate the morphogenesis of epithelial tissue, with the aim to shed light on mechanism.phentermine diseases, associated with the morphogenesis of epithelial tissue, and to develop ways of treating these diseases.

For this purpose was selected epimorphin involved in the regulation of morphogenesis of epithelial tissue (open published patent application of Japan, Publication No. 25295/94). It was found that this substance, which is the physiologically active substance containing protein, which consists of 277-289 amino acids as "crustal" protein, is synthesized mainly in mesenchymal cells. It was also found that this epimorphin has the effect of promoting the morphogenesis of epithelial tissue by acting on the cells, and the normal formation of tissue does not occur if epimorphin does not function.

With regard to the structure of epimorphin discovered that a molecule of this apomorfina can be roughly divided according to the structure 4 of the fragment (EP 0698666). That is, the polypeptide consisting of a full size apomorfina, can be divided into the following 4 fragment starting from N-Terminus: domain coiled coil (1), the functional domain (2), domain, coiled coil (3) and a hydrophobic domain at the C-end. It is assumed that among these fragments, functional domain (domain defined in apomorphine person 104-187 amino acids) involved in adhesion of cells and is closely connected with the manifestation of physiological activity of EPIM Hina (the aforementioned EP publication).

As epimorphin has effect, contributing to the normal morphogenesis, it is assumed that this substance can be used as an active ingredient in medicines for prophylactic or therapeutic treatment of diseases caused by abnormal morphogenesis, or medicines, such as agent, promotes hair growth. However, native epimorphin obtained from mammals, practically insoluble in aqueous medium, such as saline solution, which creates difficulties in the practical use of the substance as a medicinal product. There have been a few attempts to obtain new derivatives apomorfina with good solubility, while sufficiently preserving a stimulating effect on morphogenesis, characteristic of native epimorphin. For example, known is a modified form (fragment 123), obtained by removing the hydrophobic region on the C-end (open the published patent application of Japan No. 25295/1994).

It is also known that a polypeptide having a partial structure apomorfina, stimulates morphogenesis of epithelial tissue through its action on epithelial cells (international Publication WO98/22505). This peptide is soluble in an aqueous medium such as saline, and in accordance with the above-mentioned publications. is th this peptide has the ability to stimulate hair growth. However, it has not been established biological activity in the partial structure in the form of a shorter fragment.

Description of the invention

The aim of the present invention is the obtaining of oligopeptides with action, stimulating hair growth. Another object of the present invention to provide a medicinal product, preferably the agent that stimulates hair growth containing these oligopeptides as the active ingredient. The authors of the present invention have conducted extensive studies to achieve the above objectives, and as a result, they have found that some oligopeptides containing partial structure apomorfina perfectly stimulate hair growth. The present invention was made on the basis of these discoveries.

Thus, the present invention represents an Oligopeptide or its salt selected from the group consisting of

the oligopeptides of the following (1) or (2)

(1) the Oligopeptide presents

the amino acid sequence (I);

Lys-Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu

the amino acid sequence (II);

Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu

the amino acid sequence (III);

Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp

the amino acid sequence (IV);

Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln

the amino acid sequence (V);

Ser-Ile-Glu-Gln-Ser-Cys-Asp

amino acid follow what eTelestia (VI);

Ser-Ile-Glu-Gln-Ser-Cys

the amino acid sequence (VII);

Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu

the amino acid sequence (VIII);

Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu, or

the amino acid sequence (IX); or

Gln-Ser-Cys-Asp-Gln-Asp-Glu

(2) an Oligopeptide having the amino acid sequence in which one or more amino acids deleterows, substituted, or added in any one of amino acid sequences (I)to(IX), and has basically the same activity to stimulate hair growth, and Oligopeptide that represents any of the amino acid sequences (I)to(IX), and a modified Oligopeptide source of oligopeptides defined in (1) or (2)with basically the same activity to stimulate hair growth, and Oligopeptide that represents any of the amino acid sequence (I)To(IX).

In accordance with a preferred embodiment of the present invention receive Oligopeptide or its salt represented by any of the amino acid sequences (I)to(IX), or a modified Oligopeptide having basically the same activity to stimulate hair growth, and Oligopeptide that represents any of the amino acid sequences (I)to(IX). More preferred implementation includes Oligopeptide or its salt, having presented the by any of the amino acid sequence (II)-(V), or modified Oligopeptide having basically the same activity to stimulate hair growth, and Oligopeptide that represents any of the amino acid sequence (II)-(V). In accordance with the preferred embodiment of the present invention, receive Oligopeptide represented by any one of amino acid sequences (I)to(IX), and most preferably, get Oligopeptide or its salt represented by any of the amino acid sequence (I) or (IX).

An example of modified oligopeptides includes biotinylated Oligopeptide, and a more preferred example of the modified oligopeptides include Oligopeptide, whose N-terminal is connected with Biotin with a spacer or without him. Another example of modified oligopeptides include Oligopeptide, dimenisonal sulfhydryl group or a cysteine residue in the Oligopeptide.

In another aspect presents the medicinal product, including the above Oligopeptide or its physiologically acceptable salt as an active ingredient. The drug can be used as an agent for stimulating hair growth in a mammal. Additionally presents the application of the above oligopeptides or salts thereof for production of the above-mentioned Lakers the governmental funds, preferably, the agent that stimulates hair growth; and the method of stimulation of hair growth, which includes a step of introducing an effective amount of the above oligopeptides or its physiologically acceptable salt to a mammal, including humans.

Brief description of drawings

Figure 1 shows the activity of oligopeptides, which stimulates hair growth (represented by the amino acid sequence (II)) of the present invention. In the figure (A) shows the result obtained using biotinylated of oligopeptides. Square (large) shows the result obtained using oligopeptides b7, and a square (small) shows the result obtained in the control. (C) shows the result obtained using biotinylated of oligopeptides with S-S bridges. Circle (large) shows the result obtained using oligopeptides ssb7, and the circle (small) shows the result obtained in the control. The vertical axis shows the growth rate of hair, and the horizontal axis shows the day of the application.

Figure 2 shows the activity of oligopeptides, stimulating hair growth according to the present invention (biotinylated Oligopeptide with S-S bridges: ss7). The figuremeans biotinylated Oligopeptide with S-S bridges,means the result, polucen the th in the first control (the same, as the control in figure 1), andmeans the result of the second control (random 7-dimensional Oligopeptide). The vertical axis indicates the rate of hair growth, the horizontal axis shows the day of the application.

Figure 3 shows the result determine the effect of b7ΔC1, b7ΔC2, b7ΔC3, b7ΔC4, b7ΔC5, b7ΔN1, b7ΔN2, b7ΔN3 on IL-8 inducing activity. In the figure Scont denotes the result of blocking reagent, PBS denotes the result of a phosphate buffer solution, and the vertical axis shows the relative volume of the secretion of IL-8.

Figure 4 shows the results of the impact assessment of oligopeptides bk7 obtained by attaching Biotin to the N-Terminus of oligopeptides, presents the amino acid sequence (I) and oligopeptides b7 on IL-8 inducing activity. This PBS figure indicates the result of a phosphate buffer solution, and the vertical axis shows the relative volume of the secretion of IL-8.

The best option of carrying out the invention

Oligopeptide presented above amino acid sequence (I) of the present invention, an Oligopeptide consisting of the 11 C-terminal amino acid residues of the polypeptide (II)disclosed in International Publication WO98/22505. Oligopeptide presented above amino acid sequence (II), which correspond to the Oligopeptide, obtained through dellarovere amino acids (Lys) N-the end of the above amino acid sequence (I). Oligopeptide presented above amino acid sequence (III)-(VI)corresponds to the oligopeptides obtained by dellarovere one, two, three and four amino acid residues on the C-end of the aforementioned amino acid sequence (II), respectively. Oligopeptide represented by the above-mentioned amino acid sequences (VII)-(IX)corresponds to the oligopeptides obtained by dellarovere one, two and three amino acid residues at the N end of the aforementioned amino acid sequence (II), respectively. Oligopeptide represented by the above-mentioned amino acid sequences (I)to(IV), has activity, stimulating hair growth, as more specifically shown in the following examples. Not limited to any specific theory, it is possible that the C-terminal cysteine amino acid sequence of (VI) is the key for the activity.

The oligopeptides with amino acid sequence in which one or more amino acids deleterows, replaced, or inserted into one of the amino acid sequences (I)to(IX), with basically the same activity incentive is the key to hair growth, that and Oligopeptide that represents any of the amino acid sequences (I)to(IX) (further references are sometimes referred to as variant Oligopeptide)are covered by the present invention. Such modified oligopeptides based on the above oligopeptides, presents any of the above amino acid sequence (I)to(IX) or the above-mentioned variant Oligopeptide, which has basically the same activity to stimulate hair growth, and Oligopeptide that represents any of the amino acid sequences (I)to(IX), are also covered by the present invention. The person skilled in the art can easily verify that the above variant Oligopeptide and modified Oligopeptide have basically the same activity to stimulate hair growth, and Oligopeptide that represents any of the amino acid sequences (I)to(IX), using the test method described in the Examples of the present description, or through changes or modifications specified in the test method.

In the above variant of the Oligopeptide type of amino acid that can be added or replaced, is practically unlimited. Preferably it is L-amino acids. The number and position of amino acids that can be added or replaced, also practically is not limited to, for example, the number of added amino acids from 1 to 5, preferably from 1 to 3, more preferably from 1 to 2. One or more amino acids, preferably from 1 to 5, more preferably from 1 to 3 amino acids, can be added at N-end-or-end.

In the above-mentioned modified Oligopeptide term modified must be interpreted in its broadest sense, including chemical modification and biological modification. Examples of modifications include the introduction of functional groups, such as alkylation, etherification, halogenoalkane and amination, or modification of functional groups, such as oxidation, reduction, addition or deletion or insertion compounds sugars (monosaccharide, disaccharide, oligosaccharide or polysaccharide or lipid compound, phosphorylation, biotinylation. However, these modifications are not restricted by these examples.

A preferred example of the modified oligopeptides includes biotinylated Oligopeptide, and a more preferred example includes Oligopeptide, whose N-terminal is connected with Biotin with a spacer or without him. In the above Oligopeptide corresponding chemical modification can be attached to Biotin while maintaining a desired physiological activity. The way to obtain biotinylated aigopad is Yes, more detail is shown in the Examples of the present description. For the introduction of a Biotin at the N end by a spacer having an appropriate length, for example, can be used NHS-Biotin or NHS-LS-Biotin (available from Pierce).

Another preferred example of the modified oligopeptides include Oligopeptide, demersally on sulfhydryl group of the cysteine residue.

The above oligopeptides can be in free form or can be obtained in the form of additive salts of acids or additive salts of the bases. Examples of the additive salts of acids include salts of mineral acids, such as hydrochloride, sulfate, nitrate and phosphate, organic acid salts, such as para-toluensulfonate, methanesulfonate, citrate, oxalate, maleate and tartrate. Examples of the additive salts of bases include metal salts such as sodium salt, potassium salt, calcium salt, magnesium salt, ammonium salt, organic ammonium salt, such as salt methylamine and trimethylammonium salt. The Oligopeptide may form a salt with amino acids such as glycine, or may form zwitter-ion in the molecule.

Any polymers of oligopeptides, such as dimer, covered by the present invention. For example, dimenisonal Oligopeptide formed by S-S bond, is covered by the present invention. These oligopeptides can also exist in the form of a hydrate or of MES. Above all is ropeptide contain multiple asymmetric carbon atoms. Although the stereochemistry of each asymmetric carbon atom is not limited, preferably to amino acid residue would be L-amino acid. Stereoisomers such as optical isomers or diastereomers on the basis of asymmetric carbon atoms, any mixture of stereoisomers and racemates are covered by the present invention.

The oligopeptides of the present invention can be synthesized using standard chemical methods for peptide synthesis, such as solid-phase or liquid-phase technique. There are various articles about the protective groups for the amino group or the like, and the condensing agent for the condensation reaction in the synthesis of peptides and, accordingly, these articles can be referenced when conducting synthesis. In the solid-phase method can be used a variety of commercially available peptide synthesizers. Methods of introduction or removal of the protective groups, for example, in Protective Groups in Organic Synthesis, by T.W. Greene, John Wiley & Sons, Inc. 1981, etc.

Using standard biological techniques such as gene expression, the desired Oligopeptide can be obtained by constructing a recombinant vector containing the DNA sequence encoding the above Oligopeptide, obtaining microorganism (transformant), transformed by the vector, and the separation of the s and purification of oligopeptides from the culture of transformant. The method of obtaining oligopeptides not limited to these chemical and biological methods. Methods of obtaining modified oligopeptides, including chemical modification and biological modification, are well known to specialists in this field, and any of them can be used.

Oligopeptide according to this invention is suitable as an active ingredient of a medicinal product used as an agent for the stimulation of hair growth or the like. The term medicine used here in its broadest sense, including the agent for the stimulation of hair growth, which is sometimes classified as a quasi-drug, like a drug for the prophylactic or therapeutic treatment of diseases of a mammal, including humans. The term agent to stimulate hair growth, used here should be interpreted in its broadest sense, including the stimulation of the formation of hair, stimulating hair growth and preventing hair loss, and the term should not be construed in any limiting sense.

As a medicinal product according to this invention one or more substances of the above oligopeptides or their physiologically acceptable salts may themselves be used. In General, however, it is preferable to obtain and enter farmace the political composition, including one or more of the above-mentioned substances as the active ingredient with one or more pharmaceutically acceptable additives. The agent that stimulates hair growth containing one or more of the above oligopeptides, can be used in the form of external agents, such as a cream, spray, covering the mortar or plaster. The agent can be directly introduced to the target area in the form of injections. It is possible to get the agent in any form convenient for the use of the agent for the stimulation of hair growth.

For example, the above Oligopeptide as the active ingredient can be added to shampoo or conditioner, or the above Oligopeptide can be entered in liposomal capsules for the production of the drug. Compositions in the above forms are also covered by the present invention. To achieve efficient percutaneous absorption of these oligopeptides through the keratin layer of the skin in a cream, it is preferable to add the appropriate detergents, lipitorhistory substances and the like. The dose of the above-mentioned agent that stimulates hair growth, can be selected depending on the purpose, the form of the agent type of the active ingredient and the like, for Example, you can determine the dose based on the dose more accurately shown in the Examples of the present description.

Examples

the present invention will be explained in more detail in the following Examples. However, the scope of this invention are not restricted to these examples.

Example 1

Oligopeptide having the above amino acid sequence (II), were synthesized by a solid phase method using Fmoc (sometimes called RAR the following examples). Also synthesized a modified Oligopeptide (sometimes called b7 the following examples) with a modification of the Biotin at the N-end. Each of the synthesized oligopeptides was purified using high-performance liquid chromatography (HPLC), and the degree of their purity was confirmed at the level of 90% with HPLC and Mass.

The conditions of HPLC were as follows.

Column: ODS-UG-3 (Monomeric ODS, Nomura Kaguku), inner diameter 1.0 mm, length 100 mm

Measurement: room temperature (25°).

Registration: UV 214 nm, 280 nm.

Eluting solvent: gradient of solvent a and solvent B (solvent A: 0.1% triperoxonane acid; solvent B: 90% acetonitrile /0.1% of triperoxonane acid, a linear concentration gradient from 5 min after (solvent: 0%) and 55 min (solvent A: 55%).

Flow rate: 75 ml/ml

The retention time

pep7: 21,52 minutes (dimer), 20,59 minutes (monomer)

b7: 29,89 minutes (monomer), 32,85 minutes (dimer).

Example 2

Oligopeptide obtained in Example 1 was dissolved in a phosphate buffer solution (PBS) to obtain a concentration of 0.3 mg/ml, andá this solution was added the same volume of 100% ethanol to obtain a 50% ethanol/PBS solution with a concentration of 0.15 mg/ml

The Oligopeptide was made as follows.

To a solution of oligopeptides (5 ml), dissolved in PBS at a concentration of 1 mg/ml was gradually added BMH, dissolved in dimethyl sulfoxide (65 ml) under stirring to obtain a concentration of 33 μg/μl, and the reaction was carried out at 4°With during the night. Next, to the solution was added PBS (6.6 ml)and the solution of cysteine hydrochloride was dissolved in PBS (5 ml) to obtain a concentration of 5 mg/ml and mixed to obtain a solution of oligopeptides, United S-S bridges at a final concentration of 0.3 mg/ml (hereinafter in the examples above Oligopeptide obtained by crosslinking pep7, sometimes called ss7, and biotinylated ss7 - ssb7). The retention time HPLC under the above conditions 33,01 minutes. The control solution was obtained in a similar reaction with the addition of the reagents, except that instead of the solution of oligopeptides used PBS. To a solution of cross-linked oligopeptides also added the same volume of ethanol to obtain a 50% solution of ethanol/PBS with a final concentration of 0.15 mg/ml

It is known that mouse C3H and C57BL/6 have prolonged telogen phase for approximately 50 days from 45 days after birth up to 95-day. Their hair growth cycle it is easy to estimate by skin color changes, i.e. from pink in the telogen to gray or black in anagen. To assess stimulated or not the transition from telogen to anagen centuries when the Denia of oligopeptides according to this invention, was the test with these mice. Mice C57BL/6 were obtained at the age of 7 weeks (from 48 to 50 days, females) and the hair on the back (about 3×2.5 cm2) was carefully shaved with electric clippers for animals so as not to injure the skin, and it was confirmed that the skin color of the hair cycle were in the telogen phase. The solution of oligopeptides obtained as described above was applied 5 mice from each group, once a day, 5 days a week in an amount of 0.2 ml to 38 days from the start of the test. The application was carried out using a syringe without a needle. Dipeptide (Ile-Lys) and Tripeptide (Glu-Ile-Lys), in which the N-end was biotinylated were mixed and then added to cross-link agent with the reference solution.

A few people (two) observed mice twice a week to the naked eye to evaluate on a six-point scale ratio of the area of regenerated hair and square-shaved hair. Then did the photos.

Evaluation of hair growth was determined by the following method.

First, gave the following assessment depending on the ratio of the area where skin color was changed to gray or black on the area where hair has been shaved: 0-20%:1, 20-40%:2, 40-60%:3, 60-80%:4, 80-100%:5. The sum of these estimates in each group was determined as the rate of hair growth. The maximum value of the evaluation of hair growth was equal to 50 for each of the observers, and max is the maximum value of the rate of hair growth was equal to 100, since the evaluation was done by two observers. In the group, which was applied biotinylated Oligopeptide with S-S bridges, transition in anagen happened earlier on 7 or more days, than in the control group, and was apparent stimulation of hair restoration at any time, almost to the complete restoration of hair. On the other hand, in the group that used the biotinylated Oligopeptide, hair restoration was stimulated up to about 30-th day from the beginning of the test compared with the control group, similar to the S-S group. The results, shown in figure 1. The results ss7 shown in figure 2. From these results, it is concluded that the Oligopeptide, presents the amino acid sequence (II)has an activity to stimulate hair growth.

Example 3

Oligopeptides b7ΔC1, b7ΔC2, b7ΔC3, b7ΔC4 and b7ΔC5 were synthesized by dellarovere one, two, three, four or five amino acids at the C-end-b7, respectively, and then blocking sulfhydryl group. Oligopeptides b7ΔN1, b7ΔN2 and b7ΔN3 were synthesized by dellarovere one, two or three amino acids at the N end of b7. Was synthesized Oligopeptide bk7 (without blocking sulfhydryl group), presents the amino acid sequence (I), N is the end of which was connected lysine.

human Keratinocytes (NHEK cell, Colnetics available in Sanko Jyun-yaku co., Ltd.) were cultured in the medium for proliferation (Clontics), containing 30 μg/ml BPE, 0.1 ng/ml human EGF, 5 μg/ml insulin, 0.5 μg/ml hydrocortisone, 50 μg/ml gentamicin and 50 ng/ml amphotercin. These cells were re-suspended at a concentration of 1 x 104cells/ml in the environment in which EGF, insulin and hydrocortisone were removed from the medium for proliferation, and then 100 μl of the suspension was placed in each well of 96-well plate. Simultaneously with the application of cell suspension was added 5 μl of 1 mg/ml of oligopeptides to obtain a final concentration of 50 µg/ml After cultivating for 16-20 hours, the amount of IL-8 in the culture supernatant was measured with an ELISA kit (ENDOGEN). There was obtained a good correlation between the inducing activity of the secretion of IL-8 by NHEK cells and activity, stimulating hair growth (table 1).

The results obtained in the evaluation inducing activity of oligopeptides b7ΔC1, b7ΔC2, b7ΔC3, b7ΔC4, b7ΔC5, b7ΔN1, b7ΔN2 and b7ΔN3 on IL-8 are shown in Figure 3. When one or more amino acids have deleterule N-end, the amount of secreted IL-8 decreased slightly, at a time when one or more amino acids have been deleterow on the C-end, the amount of secreted IL-8 was maintained until dellarovere 4 amino acids. When DeleteMovie 5 amino acids with kreditovanie the amount of IL-8 were significantly decreased. These results indicate that we can expect almost the same activity, stimulating hair growth, as the oligopeptides b7, when DeleteMovie up to 4 amino acids on the C-end, or when DeleteMovie up to 3 amino acids at N-end. As shown in Figure 4, the 11-dimensional Oligopeptide bk7 had almost the same IL-8 inducing activity, and Oligopeptide b7.

Example 4

N-Terminus of oligopeptides pep7 was biotinylated using NHS-Biotin or NHC-Biotin (Pierce) according to instructions of the accompanying manual. When used NHS-Biotin, -O-CO-(CH2)4-(13,5) was introduced as a spacer between the N-end and Biotin, and when used NHS-LC-Biotin, -O-CO-(CH2)5-NH-CO-(CH2)4-(22,4) was introduced as a spacer between the N-end and Biotin. The amount of secretion of IL-8 was determined using biotinylated oligopeptides as in Example 3. These oligopeptides have almost the same IL-8 inducing activity, and Oligopeptide b7. These results show that the Oligopeptide, N-end biotinylated directly, has almost the same activity to stimulate hair growth, and Oligopeptide in which the N-end biotinylated through a spacer, and both are more active compared to the Oligopeptide, which was not introduced Biotin.

<> Industrial applicability

Oligopeptide according to this invention has activity to stimulate hair growth and is effective as an active ingredient in a medicinal product, such as an agent that stimulates hair growth.

1. Oligopeptide or its salt selected from the group consisting of oligopeptides of the following (1) and (2):

(1) the Oligopeptide presented

the amino acid sequence (I):

Lys-Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu

the amino acid sequence (II):

Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu

the amino acid sequence (III):

Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp

the amino acid sequence (IV):

Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln

the amino acid sequence (V):

Ser-Ile-Glu-Gln-Ser-Cys-Asp

amino acid sequence of (VI):

Ser-Ile-Glu-Gln-Ser-Cys

the amino acid sequence (VII):

Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu

the amino acid sequence (VIII):

Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu or

the amino acid sequence (IX):

Gln-Ser-Cys-Asp-Gln-Asp-Glu,

(2) an Oligopeptide having the amino acid sequence obtained by dellarovere C - or N - Termini of one or more amino acid any amino acid sequence (I)to(IX), and having basically this is e activity to stimulate hair growth, that and Oligopeptide that represents any of the amino acid sequences (I)to(IX), and a modified Oligopeptide representing biotinylated or demersally on sulfhydryl group of the cysteine residue Oligopeptide-based oligopeptides, defined in (1) or (2)with basically the same activity to stimulate hair growth, and Oligopeptide that represents any of the amino acid sequences (I)to(IX).

2. Oligopeptide or its salt according to claim 1, where the modified Oligopeptide is the biotinylated Oligopeptide.

3. Oligopeptide or its salt according to claim 2, where the modified Oligopeptide is an Oligopeptide, where N is the end connected with Biotin with a spacer or without it.

4. Oligopeptide or its salt according to claim 1, where the modified Oligopeptide is an Oligopeptide, dimeric by sulfhydryl group of the cysteine residue in the amino acid sequences described above.

5. Oligopeptide or its salt represented by any of the amino acid sequences (I)to(IX), according to claim 1, and a modified Oligopeptide representing biotinylated or demersally on sulfhydryl group of the cysteine residue Oligopeptide, which has basically the same activity to stimulate hair growth, and Oligopeptide represented by l is the battle of the amino acid sequences (I)to(IX).

6. Oligopeptide or its salt according to claim 5, where the modified Oligopeptide is the biotinylated Oligopeptide.

7. Oligopeptide or its salt according to claim 6, where the modified Oligopeptide is an Oligopeptide, where N is the end connected with Biotin with a spacer or without it.

8. Oligopeptide or its salt according to claim 5, where the modified Oligopeptide is an Oligopeptide, dimeric sulfhydryl group of the cysteine residue in the above-described amino acid sequences.

9. Oligopeptide or its salt represented by any of the amino acid sequences (I) to (IX) under item 1.

10. Drug for stimulating hair growth, comprising as an active ingredient Oligopeptide according to any one of claims 1 to 8 or its physiologically acceptable salt.

11. Method of promoting hair growth, comprising the stage of introducing an effective amount of oligopeptides according to any one of claims 1 to 8 or its physiologically acceptable salt to a mammal, including humans.



 

Same patents:

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

The invention relates to the field of biotechnology, specifically to peptides active in the attachment, respectiveiy and detaching cells, and can be used to study activity in the attachment of cells, mediated by different proteins of the extracellular matrix, to develop peptides, adhesion, and also for medicinal purposes

The invention relates to peptide compositions with delayed release, representing a compound (I) containing the compound (A) formula

and the polymer containing lactide links, glycolide links and links tartaric acids - which are found in the polymer at the next sootnoshenii: lactide units constitute from about 71% to about 73%, glycolide links from about 26% to about 28%; and the parts of tartaric acid from about 1% to 3%, and the amino group of the compound (a) relate ionic bond with the carboxyl groups of the acid units of the polymer; the particles of compound I, an average size of from about 10 microns to about 100 microns; pharmaceutical composition with delayed release and two methods of treatment of various diseases, including the introduction to the patient an effective amount of compound A, or microparticles

The invention relates to medicine and relates to methods of treatment of tumors and metastases using a combination of antiangiogenic therapy and immunotherapy

The invention relates to new peptides having the amino acid sequence of the 9-55 amino acid residues, comprising the amino acid sequence FTLASAETT (SEQ ID NO:l), pharmaceutical compositions for treatment of autoimmune diseases, containing one or more of these peptides and a pharmaceutically acceptable carrier

The invention relates to a LHRH antagonists - compounds of General formula Iin which a represents acetyl or 3-(4-forfinal)propionyloxy group Xxx1mean D-Nal(1) or D-Nal(2), Xxx2-Xxx3mean D-Cpa-D-Pal(3) or a simple link, Xxx4means Ser, Xxx5means N-Me-Tyr, Xxx6mean D-Hci or a residue of D-amino acids of General formula (II)

where n means the number 3 or 4, a R1means a group of the General formula IIIwhere R denotes an integer from 1 to 4, R2means hydrogen or alkyl group, and R3means unsubstituted or substituted aryl group or heteroaryl group, or R1mean 3-amino-1,2,4-triazole-5-carbonyl group,Xxx7means Leu or Nle, Xxx8means Arg or Lys(iPr), Xxx9means Pro and Xxx10means A1A or Sar, and their salts with pharmaceutically acceptable acids: process for the preparation of these compounds, pharmaceutical compositions having the properties of an LHRH antagonist, comprising as an active narushenie compounds according to the invention has a high solubility in water

The invention relates to biotechnology and can be used to obtain a polypeptide with IL-10 similar properties

The invention relates to medicine, in particular for the treatment of male pattern baldness type

The invention relates to medicine, namely to methods for treating alopecia

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

FIELD: medicine, first aid, anesthesiology, resuscitation, surgery.

SUBSTANCE: along with conventional medicinal preparations applied to treat shock one should introduce crystalloids into central vein in certain sequence: 7.5% and 0.9%-sodium chloride solution, 5%-glucose solution, and, also, infucol and similar-group plasma; after stabilizing arterial pressure one should introduce, additionally, either mildronate, or dalargin at certain dosages. The present innovation enables to restore the volume of extracellular liquid in the shortest period of time at decreased volume of infusion that, in its turn, favors to remove shock and prevent other possible further complications.

EFFECT: higher efficiency.

3 ex

FIELD: medicine, surgery.

SUBSTANCE: one should introduce solution "Rheamberin 1.5%"intravenously with infusomates at the dosage of 5 ml/kg/d during the next 5 d. Then, 1-2 h later, one should infuse intravenously "Dalargin" at 30 mg/kg/d dissolved in 60 ml 0.9% sodium chloride at the rate of 120 ml/h for 5 d. Additionally, 4-5 h after "Dalargin" injection it is necessary to perform daily intravenous He-Ne laser irradiation of blood beginning since the first day at wave length being 0.63 mcm, power 1 mW, exposure 50-60 min, course lasts for 5 d. The method enables to interrupt intestinal paresis in case of vertebral traumas and wounds in earlier terms.

EFFECT: higher efficiency of therapy.

1 ex

FIELD: chemistry of peptides, medicine, oncology, pharmaceutical chemistry.

SUBSTANCE: invention relates to the development of medicinal agent of peptide nature eliciting an antitumor effect and can be used in treatment of endocrine and hormone-dependent tumors. Agent represents peptide of the general formula: . Invention provides enhancement of the therapeutic effect and reducing toxicity.

EFFECT: valuable medicinal properties of agent.

3 cl, 4 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: peptide of the following formula: X-Pro-Gly-P, where X = Thr-Lys-Pro-Arg-; Lys-pro-Arg-; pro-Arg-; Arg-, being of untiulcerous activity. They should be applied at intraperitoneal injection at the dosage of 0.58-3.20 mcM g/kg for preventing and treating ulcers of gastro-intestinal tract.

EFFECT: higher efficiency and prophylaxis.

4 dwg, 5 ex

FIELD: genetic engineering, medicine.

SUBSTANCE: invention relates to T-cell receptor sequence being detected in patients with extended sclerosis and is useful in diagnosis and therapy. Oligonucleotide including sequence which represents or is derived from 5'-CTAGGGCGGGCGGGACTCACCTAC-3' or nucleotide sequence being fully complementary thereto. Oligonucleotide together with nuclear acid including nearly 15-30 oligonucleotides, which doesn't comprise oligonucleotide sequence and presents in region from Vβ to Jβ of Vβ13.1 gene in T-cell Vβ13.1-subgroup, wherein oligonucleotide and nuclear acid sequences don't present in the same chain of pair sequences of Vβ13.1 gene, is used in Vβ13.1 gene part amplification. In method for detection of LGRAGLTY motive, which is present in T-cell receptors of T-cell Vβ13.1-subgroup, oligonucleotide is used in combination with labeling particle. Once LGRAGLTY motive is detected, development monitoring and treatment are carried out by removing of LGRAGLTY motive-containing peptide.

EFFECT: simplified methods for detection of LGRAGLTY motive in T-cell receptors and treatment of patients with extended sclerosis.

21 cl, 7 dwg, 3 tbl, 3 ex

FIELD: pharmaceutical chemistry, medicine.

SUBSTANCE: in the suggested composition one should apply heptapeptide of Met-Glu-His-Phe-Pro-Gly-Pro sequence (heptapeptide A) for treating ischemic insult due to introducing 2 drops of compositions into each nasal canal 5-6 times daily for 10 d at disease of average severity degree, and in case of severe degree - per 3 drops of the present composition into each nasal canal 7 times daily for 10 d. The present innovation provides increased efficiency at decreased concentration of heptapeptide without any side effects.

EFFECT: higher efficiency of therapy.

2 cl, 6 dwg, 8 ex, 5 tbl

FIELD: medicine, phthisiology, anesthesiology.

SUBSTANCE: during the day of operation one should perform autohemotransfusion, then introduce epocrine intravenously by drops at the dosage of 50-200 U/kg patient's body weight; next day after interference one should inject epocrine subcutaneously at the dosage of 25-100 U/kg; at hematocrit level being below 35% 48 h after operation it is necessary to repeat subcutaneous injection of the above-mentioned preparation at the dosage not exceeding 50 U/kg. The present innovation favors hemopoiesis stimulation in postoperational period, that, in its turn, accelerates postoperational rehabilitation in patients of this group and enables, also, to avoid allotransfusions being dangerous because of immunoconflicting reactions.

EFFECT: higher efficiency of compensation.

1 ex, 1 tbl

FIELD: medicine, cardiology.

SUBSTANCE: the suggested method should be performed at the background of medicinal therapy with preparations out of statins group, tevetene, polyoxidonium and conducting seances of plasmapheresis by removing 800 ml plasma twice weekly with N 5 due to additional intramuscular injection of immunophan 0.005%-1.0 with N 10 and fluimucyl 300 mg intravenously daily with N 5-10, total course of therapy lasts for 2 mo. The method provides modulation of leukocytic functional activity, moreover, due to altered cytokine profile and, thus, through disintegration of protein-lipid complexes participating in the development of atherosclerotic platelets.

EFFECT: higher efficiency of therapy.

3 ex

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