Composition of prolonged effect and method for preparing this composition

FIELD: chemistry of peptides, medicine, drugs, pharmacy.

SUBSTANCE: invention relates to pharmaceutical compositions containing polymer of lactic and glycolic acids with the ratio of average molecular mass and medium-numbered molecular mass about 1.90 or less, or its salt and physiologically active peptide as agonist of LH-RH of formula: 5-oxo-Pro-His-Trp-Ser-Tyr-Y-Leu-Arg-Pro-Z (I) wherein Y means D-Leu, D-Ala, D-Trp, D-Ser (tBu), D-2-Nal or D-His; Z means HN-C2H5 or Gly-NH2 or its salt, in particular, peptide of the formula: 5-oxo-Pro-His-Trp-Ser-Tyr-D-Leu-Leu-Arg-Pro-NH-C2H5 (II) or its acetate, and to polymer of lactic and glycolic acids with the ratio of average molecular mass and medium-numbered molecular mass about 1.90 or less. Also, invention relates to a method for preparing indicated polymer and to a microsphere containing indicated polymer and peptide of the formula (II), and to a medicinal agent used in prophylaxis or treatment of prostate cancer, prostatomegaly, endometriosis, uterus myoma, metrofibroma, premature sexual maturation and dysmenorrhea, or for a contraceptive agent comprising indicated microsphere, and to methods for prophylaxis or treatment of indicated diseases. Invention provides preparing compositions with delayed release and containing physiologically active substance of peptide nature and providing the stable rate of release of active substance for the prolonged period (up to 1 month) in inhibition of high initial release of indicated active substance.

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

37 cl, 4 tbl, 9 ex

 

The technical field

The present invention relates to the preparation of prolonged action, containing a physiologically active substance, and the way to obtain this drug.

Background of the invention

In the patent application of Japan No. 60-100516 described microcapsule prolonged action with water-soluble drug, which is a particle with an average diameter of 2 to 200 μm, containing water-soluble drug dispersed in the matrix polymer of lactic and glycolic acid with an average molecular weight of about 5000-200000, including about 100-50 wt.% lactic acid and about 0-50 wt.% glycolic acid, which is obtained by the method of drying in the water.

In the patent application of Japan No. 62-201816 described microcapsule prolonged action, characterized in that the viscosity of the emulsion water in oil when receiving emulsion water in oil in water was adjusted to approximately 150-10000 CP, and the method of obtaining the specified microcapsules.

In the patent application of Japan No. 62-54760 described biodegradable ester policecarauctions acid, representing the copolymer or polymer with an average molecular weight of about 2000-50000 containing water-soluble hydroxycarboxylic acid number of less than 0.01 mol/100 g in terms of monobasic acid, and intended for injection m is croupal prolonged action, containing the specified polymer.

In the patent application of Japan No. 61-28521 described polymer of lactic and glycolic acid with an average molecular weight of about 5000-30000, not containing catalyst, which is characterized dispersible (according to the results of gel chromatography) of about 1.5-2 and contains about 50-95 wt.% lactic acid and 50-5 wt.% glycolic acid, and pharmaceutical preparation containing the specified polymer as the base.

In the patent application of Japan No. 6-192068 describes a method for microcapsules with prolonged action, which is that the microcapsule is heated at a temperature above the glass transition temperature of the polymer, in which the corresponding particles in the microcapsules do not stick to each other.

In the patent application of Japan No. 4-218528 described method of cleaning biodegradable aliphatic polyester, which is that the biodegradable aliphatic polyester containing low molecular weight polymer with a molecular weight of 1000 or less, dissolved in an organic solvent, water is added to precipitate the polymer mass and remove the low molecular weight polymer with a molecular weight of 1000, and water is added in the amount of 50-150 parts (volumes) per 100 parts of organic solvent.

The purpose of the invention

The present invention relations is seeking to prolonged drug action, which does not contain gelatin, containing a physiologically active substance in large quantities and providing a stable rate of release for about one month during the suppression of the high initial release of physiologically active substances.

A brief statement of the substance of the invention

In order to solve the above problem, the authors of the present invention conducted a comprehensive survey, which was established drug prolonged action, containing a large number of physiologically active substances and do not contain gelatin, which suppresses the high initial release of physiologically active substances and provides a stable rate of release for about one month, with the specified drug as the basis used PLGA copolymer with a ratio of the average molecular weight and srednekamennogo molecular weight of about 1,90 or less or a polymer of lactic and glycolic acid with an average molecular weight from about 11600 to about 140,000 or its salt.

Thus, objects of the present invention are:

(1) composition of prolonged action, containing a polymer of lactic and glycolic acid with a ratio of the average molecular weight and srednekamennogo molecular weight of the olo 1,90 or less or its salt and a physiologically active substance;

(2) the composition of prolonged action described in paragraph (1), in which the physiologically active substance is a physiologically active peptide;

(3) the composition of prolonged action described in paragraph (2), in which the physiologically active substance is derived LH-RH;

(4) the composition of prolonged action described in paragraph (1), in which the polymer of lactic and glycolic acid has an average molecular weight of from about 3,000 to about 100,000;

(5) the composition of prolonged action described in paragraph (4), in which the polymer of lactic and glycolic acid has an average molecular weight of from about 8,000 to about 15,000;

(6) the composition of prolonged action described in paragraph (1), in which the low molecular weight fraction of the polymer of lactic and glycolic acids with a mass of about 3000 or below is about 9% or less;

(7) the composition of prolonged action described in paragraph (6), in which the low molecular weight fraction of the polymer of lactic and glycolic acids with a mass of about 3000 or below is from about 3% to about 9%;

(8) the composition of prolonged action described in paragraph (1), in which the specified polymer is characterized by a molar ratio of lactic acid and glycolic acid from 100:0 to 40:60;

(9) the composition of prolonged action described in paragraph (1), in which the specified polymer is characterized by a molar ratio of lactic acid and glycolic acid 70:30 to 80:20;

(10) the composition of prolonged action described in paragraph (3), which is derived LH-RH peptide is expressed by the formula

5-oxo-Pro-His-Trp-Ser-Tyr-Y-Leu-Arg-Pro-Z

where Y represents DLeu, DAla, DTrp, DSer (tBu), D2Nal or DHis (ImBzl), and Z denotes HN-C2H5or Gly-NH2or its salt;

(11) the composition of prolonged action described in paragraph (3), which is derived LH-RH peptide is expressed by the formula

5-oxo-Pro-His-Trp-Ser-Tyr-DLeu-Leu-Arg-Pro-NH-C2H5,

or its acetate;

(12) the composition of prolonged action described in paragraph (3), which is derived LH-RH or its salt is in an amount of from about 5% (wt./wt.) to about 24% (wt./wt.);

(13) the composition of prolonged action described in paragraph (1), in which the physiologically active substance or its salt are soluble in water or water-soluble;

(14) the composition of prolonged action described in paragraph (1), which is designed for injection;

(15) the composition of prolonged action described in paragraph (1), which releases a physiologically active substance or its salt, at least within two weeks.

(16) the composition of prolonged action described in paragraph (1), to ora does not contain the substance, retaining the medicinal product;

(17) the composition of prolonged action described in paragraph (1), which does not contain gelatin;

(18) the production method of the composition of prolonged action described in paragraph (1), which involves the removal of solvent from a mixed solution containing a physiologically active substance or its salt and a polymer of lactic and glycolic acid with a ratio of the average molecular weight and srednekamennogo molecular weight of about 1,90 or less or its salt;

(19) the method described in paragraph (18), which comprises mixing and dispersing a physiologically active substance or its salt in solution in an organic solvent containing the polymer of lactic and glycolic acid with a ratio of the average molecular weight and srednekamennogo molecular weight of about 1,90 or less or its salt and then removing the organic solvent;

(20) the method described in paragraph (19), in which the physiologically active substance or its salt is used in the form of an aqueous solution containing a physiologically active substance or its salt;

(21) the pharmaceutical preparation containing the composition of the prolonged action described in paragraph (1);

(22) a drug for preventing or treating prostate cancer, prostatomegaly, endometriosis, mi is we uterus, metrofibroma, premature puberty, dysmenorrhea or breast cancer or a contraceptive pill, which contains the composition of the prolonged action described in paragraph (3);

(23) a drug to prevent recurrence of breast cancer after the operation for premenopausal breast cancer, which contains the composition of the prolonged action described in paragraph (3);

(24) a method of preventing or treating prostate cancer, prostatomegaly, endometriosis, fibroids, metrofibroma, premature puberty and dysmenorrhea or use of contraceptives, which includes an introduction to the mammal an effective dose of the composition of prolonged action described in paragraph (3);

(25) a method for preventing recurrence of breast cancer after the operation for premenopausal breast cancer, which comprises the administration to a mammal an effective dose of the composition of prolonged action described in paragraph (3);

(26) a method of obtaining a polymer of lactic and glycolic acid having an average molecular weight of from about 8,000 to about 15,000 and the ratio of the average molecular weight and srednekamennogo molecular weight of about 1,90 or less, or its salt, which comprises adding water to the organization is practical solvent, containing a polymer of lactic and glycolic acid with an average molecular weight from about 5,000 to about 15,000, with a ratio of less 5-50 parts (volumes) per 100 parts of organic solvent;

(27) a method of obtaining a polymer described in paragraph (26), in which the organic solvent is hydrophilic;

(28) a method of obtaining a polymer described in paragraph (27), in which the hydrophilic organic solvent is acetone;

(29) a method of obtaining a polymer described in paragraph (26), in which the ratio of water to 100 parts of organic solvent is from about 10 to about 45 parts (volumes);

(30) a method of obtaining a polymer described in paragraph (26), in which the ratio of water to 100 parts of organic solvent is about 40 parts (volumes);

(31) a polymer of lactic and glycolic acid having an average molecular weight of from about 8,000 to about 15,000 and the ratio of the average molecular weight and srednekamennogo molecular weight of about 1,90 or less, or its salt;

(32) the use of the polymer of lactic and glycolic acids described in paragraph (31), or its salt to obtain compositions with prolonged action, does not contain gelatin;

(33) microsphere containing polymer of lactic and glycolic acid with an average molecular weight from about 11600 to the Colo 14000 or its salt and a derivative of LH-RH or its salt and does not contain gelatin;

(34) microsphere described in paragraph (33), which is derived LH-RH or its salt is a peptide expressed by the formula

5-oxo-Pro-His-Trp-Ser-Tyr-Y-Leu-Arg-Pro-Z

where Y represents DLeu, DAla, DTrp, DSer (tBu), D2Nal or DHis (ImBzl), and Z denotes HN-C2H5or Gly-NH2or its salt;

(35) microsphere described in paragraph (33), which is derived LH-RH or its salt is a peptide expressed by the formula:

5-oxo-Pro-His-Trp-Ser-Tyr-DLeu-Leu-Arg-Pro-NH-C2H5,

or its acetate;

(36) microsphere described in paragraph (33), which contains a derivative of LH-RH or its salt in an amount of from about 5% (wt./wt.) to about 24% (wt./wt.);

(37) microsphere described in paragraph (33), which is a microcapsule;

(38) microsphere described in paragraph (33), which releases derived LH-RH or its salt, at least for more than 2 weeks;

(39) a drug for preventing or treating prostate cancer, prostatomegaly, endometriosis, fibroids, metrofibroma, premature puberty and dysmenorrhea or contraceptive pill, which contains the microsphere described in paragraph (33);

(40) a drug to prevent recurrence of breast cancer after the operation for premenopausal breast cancer, which contains the microsphere described in paragraph (33);

(41) the way PR is in preventing or treating prostate cancer, prostatomegaly, endometriosis, fibroids, metrofibroma, premature puberty and dysmenorrhea or use of contraceptives, which includes an introduction to the mammal an effective dose of the microspheres described in paragraph (33);

(42) a method for preventing recurrence of breast cancer after the operation for premenopausal breast cancer, which comprises the administration to a mammal an effective dose of the microspheres described in paragraph (33);

Detailed description of the invention

Physiologically active substance used in the present invention has no specific limitation, if it is pharmaceutically useful ones or peptide compound. Examples of acceptable ones connections include agonist, antagonist and a compound having inhibitory enzyme activity. In addition, the preferred peptide compound is a physiologically active peptide. To achieve this purpose, suitable physiologically active peptides with a molecular weight of from about 3,000 to about 40,000, preferably from about 4000 to about 30,000, more preferably from about 5,000 to about 200,000.

Examples of physiologically active peptides include releasing hormone luteinizing hormone (LH-RH), insulin, somatostatin, somatotropin, a hormone-releasing hormone, gormo the and growth (GH-RH), prolactin, erythropoietin, cortical hormone of the adrenal cortex, melanocytestimulating hormone releasing hormone, thyroid hormone, the hormone that stimulates the thyroid gland, luteinizing hormone, follicle stimulating hormone, vasopressin, oxytocin, calcitonin, gastrin, secretin, pancreozymin, cholecystokinin, angiotensin, placental lactogenic human chorionic gonadotropin human enkephalin, endorphin, biotropin, tuftsin, thymopoietin, thymosin, timtiman, humoral factor thymic factor blood thymus, tumor necrosis factor, kolonieobrazuyushchei factor, motilin, dynorphin, bombezin, neurotensin, cerulein, bradykinin, the amplification factor of the urinary tract, the factor nerve growth, growth stimulator cells, neurotrophic factors, peptides having antagonistic activity against endothelin, and their derivatives, fragments and derivatives of these fragments.

Physiologically active substance used in the present invention may be the product itself or its pharmacologically acceptable salt.

If the physiologically active substance has a basic group such as amino group, examples of the salts include salts with inorganic acids (also referred to as free inorganic acids (e.g. carbonic acid, bigolina acid, hydrochloric Ki the lot, sulfuric acid, nitric acid, boric acid, and the like) and organic acids (also referred to as free organic acids (e.g. succinic acid, acetic acid, propionic acid, triperoxonane acid and the like); when the physiologically active substance has an acidic group such as carboxyl group and the like, examples of the salts include salts with inorganic bases (also referred to as free inorganic bases) (for example, alkali metal such as sodium, potassium and the like, alkaline earth metal such as calcium, magnesium and the like) and organic bases (also referred to as free organic basis) (for example, organic amines such as triethylamine and the like, basic amino acids such as arginine and the like). In addition, the physiologically active peptide may form a complex compound of the metal (e.g. copper complex, a complex of zinc, and the like).

Preferred examples of physiologically active peptides include derivatives of LH-RH or their salts, which can effectively treat hormonal diseases, in particular cancer is caused by sex hormone (e.g., prostate cancer, uterine cancer, breast cancer, pituitary tumor, and things like the s) diseases caused by sex hormone, such as prostatomegaly, endometriosis, uterine fibroids, premature puberty, dysmenorrhea, amenorrhea, premenstrual syndrome, multi-chambered ovary, and the like, provide contraception (or restore activity after discontinuation of contraceptives and infertility treatment), prevention of relapse of breast cancer after the operation for premenopausal breast cancer. Moreover, these examples include derivatives of LH-RH or their salts, to effectively treat benign and malignant tumors are not caused by sex hormone, but is sensitive to LH-RH.

Typical examples of derivatives of LH-RH or their salts include the peptides described in the Treatment with GnRH analogs: Controversies and perspectives (The Parthenon Publishing Group Ltd.), published in 1996, applications for Japan patent No. 3-503165, 3-101695, 7-97334 and 8-259460.

Examples of derivatives of LH-RH include an agonist of LH-RH and antagonists of LH-RH. As antagonists of LH-RH can be used, for example, physiologically active peptides of the General formula [I]

X-D2Nal-D4ClPhe-D3Pal-Ser-A-B-Leu-C-Pro-DAlaNH2

[where X is N(4H2-furoyl) Gly or NAc, a represents a residue selected from NHeTyr, Tyr, Aph(Atz) and NMeAph(Atz), means a residue selected from DLys(Nic), DCit, DLis (AzaglyNic), DLis(AzaglyFur), DhArg(Et2), DAph(Atz) and DhCi, and means Lys(Nisp), Arg or hArg(Et2 )],

or their salts.

As an agonist of LH-RH can be used, for example, physiologically active peptides expressed by the General formula [II]

5-oxo-Pro-His-Trp-Ser-Tyr-Y-Leu-Arg-Pro-Z

[where Y denotes a residue selected from DLeu, DAla, DTrp, DSer(tBu), D2Nal and DHis(ImBzl), and Z signifies NH-C2H5or Gly-NH2]

or their salts. In particular, for this purpose suitable peptide, in which Y represents DLeu and Z signifies NH-C2H5(i.e. peptide And expressed by the formula 5-oxo-Pro-His-Trp-Ser-Tyr-DLeu-Leu-Arg-Pro-NH-C2H5; leiprorelina) or its salt (e.g. acetate).

These peptides can be obtained by methods described in the above publications and published patents or similar methods.

Abbreviations used in the present description of the invention, have the following meanings:

AbbreviationName
N(4H2-furoyl)GlyThe remainder of the N-tetrahydropalmatine
NAcN-acetyl group
N2Nalthe remainder of D-3-(2-naphthyl)alanine
D4ClPhethe remainder of D-3-(4-chloro)phenylalanine
D3Palthe remainder of D-3-(3-pyridyl)alanine
NMeTyrthe remainder of the N-methyltyrosine
Aph(Atz) N-[5'-(3'-amino-1'H-1',2',4'-triazolyl)]phenylalanine
NmeAph(Atz)the remainder of the N-methyl[5'-(3'-amino-1'H-1',2',4'-triazolyl)]phenylalanine
DLys(Nic)the residue is D-e-N-nicotinoyl)lysine
Dcitthe remainder of the D-citrulline
DLys(AzaglyNic)the remainder of D-(isapiconnection)lysine
DLys(AzaglyFur)the remainder of D-(utilityperson)lysine
DhArg(Et2)the remainder of D-(N,N'-diethyl)homoarginine
DAph(Atz)the remainder of D-N-[5'-(3'-amino-1'H-1',2',4'-triazolyl)]phenylalanine
DhCithe remainder of the D-homocitrullinuria
Lys(Nisp)the residue (e-N-isopropyl)lysine
hArg(Et2)the remainder (N,N'-diethyl)homoarginine

As for the other acids, the abbreviations used correspond to the abbreviations approved (IUPAC-IUB) (European Journal of Biochemistry, Vol. 138, pp 9-37 (1984)), or conventional abbreviations adopted in this area. In addition, if the amino acid has an optical isomer, such an acid is defined as L-amino acid except where otherwise noted.

When carrying out the present invention it is preferable to use a polymer of lactic and glycolic acids having a ratio of average mol the molecular weight and srednekamennogo molecular weight, approximately 1,90 or less.

The polymer of lactic and glycolic acids can represent salt. Examples of such salt include salts with inorganic bases (e.g. alkali metal such as sodium, potassium and the like, alkaline earth metal such as calcium, magnesium and the like) or organic bases (e.g. organic amines such as triethylamine and the like, and basic amino acids such as arginine and the like), salts with transition metals (e.g. zinc, iron, copper and the like) and a complex salt.

The molar ratio of lactic acid and glycolic acid polymer is preferably from about 100/0 to about 40/60, more preferably from about 70/30 to about 80/20.

The ratio of optical isomers, representing the D-isomer/L-isomer (mol/mol.%), in lactic acid, which is one of the minimum structural units in the polymer of lactic and glycolic acid", preferably is from about 75/25 to about 25/75. In particular, often used the ratio of the D-isomer/L-isomer (mol/mol.%) in the range of about 60/40 to about 30/70.

The average molecular mass of the polymer of lactic and glycolic acids" is usually 3,000 to about 100,000, preferably from about 3,000 to about 50,000, most preferably from about 8,000 to about 15,000.

<> Low molecular weight fraction with a molecular weight of about 3,000 or less of a polymer of lactic and glycolic acid" is preferably about 9% or less, more preferably about 3 to 9% or less.

In addition, the polymer of lactic and glycolic acids according to the present invention has the ratio of the average molecular weight and srednekamennogo molecular weight of about 1,90 or less, preferably from about 1.40 to about 1,90, more preferably from about 1.45 to about 1.80.

Preferred examples include:

(1) a polymer of lactic and glycolic acid having an average molecular weight of from about 3,000 to 100,000 and the ratio of the average molecular weight and srednekamennogo molecular weight of about 1,90 or less;

(2) a polymer of lactic and glycolic acid having an average molecular weight of from about 3,000 to 50,000 and a ratio of the average molecular weight and srednekamennogo molecular weight of about 1,90 or less;

(3) a polymer of lactic and glycolic acid having an average molecular weight of from about 8000 to 15000 and the ratio of the average molecular weight and srednekamennogo molecular weight of about 1,90 or less;

(4) the composition of prolonged action, as described in paragraphs (1) to (3), in which the low molecular weight fraction with a molecular weight of about 3000 or less of a polymer of lactic and glycolic acid is about 9% is less;

(5) the composition of prolonged action, as described in paragraphs (1) to (3), in which the low molecular weight fraction with a molecular weight of about 3000 or less of a polymer of lactic and glycolic acid is from about 3% to about 9%.

More preferable examples include:

(6) the polymer of lactic and glycolic acid having an average molecular weight of from about 3,000 to 100,000 and the ratio of the average molecular weight and srednekamennogo molecular weight of from about 1.40 to about 1,90;

(7) the polymer of lactic and glycolic acid having an average molecular weight of from about 3,000 to 50,000 and a ratio of the average molecular weight and srednekamennogo molecular weight of from about 1.40 to about 1,90;

(8) a polymer of lactic and glycolic acid having an average molecular weight of from about 8000 to 15000 and the ratio of the average molecular weight and srednekamennogo molecular weight of from about 1.40 to about 1,90;

(9) the composition of prolonged action, as described in paragraphs (6) to (8), in which the low molecular weight fraction with a molecular weight of about 3000 or less of a polymer of lactic and glycolic acid is about 9% or less;

(10) the composition of prolonged action, as described in paragraphs (6) to (8), in which the low molecular weight fraction with a molecular weight of about 3000 or less of a polymer of lactic and glycolic acid is from about 3% dookola 9%.

The most preferable examples include:

(11) the polymer of lactic and glycolic acid having an average molecular weight of from about 3,000 to 100,000 and the ratio of the average molecular weight and srednekamennogo molecular weight of from about 1.45 to about 1.80;

(12) a polymer of lactic and glycolic acid having an average molecular weight of from about 3,000 to 50,000 and a ratio of the average molecular weight and srednekamennogo molecular weight of from about 1.45 to about 1.80;

(13) the polymer of lactic and glycolic acid having an average molecular weight of from about 3000 to 15000 and the ratio of the average molecular weight and srednekamennogo molecular weight of from about 1.45 to about 1.80;

(14) the composition of prolonged action, as described in paragraphs (11) - (13), in which the low molecular weight fraction with a molecular weight of about 3000 or less of a polymer of lactic and glycolic acid is about 9% or less;

(15) the composition of prolonged action, as described in paragraphs (11) - (13), in which the low molecular weight fraction with a molecular weight of about 3000 or less of a polymer of lactic and glycolic acid is from about 3% to about 9%.

In addition, it is possible to use a polymer of lactic and glycolic acid with an average molecular weight from about 11600 to about 14000 or its salt.

The term "average molecular weight" and "srednekislye forefront of the popular weight" in the present description of the invention, the mean molecular weight in terms of the molecular weight of polystyrene, the measured gel chromatography (GPC) using as a standard substance of the ten types of monodisperse polystyrene (GPC1) with an average molecular weight 397000, 189000, 98900, 37200, 17100, 9490, 5870, 2500, 1050 and 495.

In addition, the content of low molecular weight fraction of polymer with a molecular weight of about 3000 or less corresponds to the fraction with molecular weight of about 3000 or less on the pattern of distribution of the average molecular weight obtained by performing the above GPC measurement. In particular, determine the area under the curve portion corresponding to a molecular weight of about 3,000 or less, relative to the area under the curve pattern of distribution of the calculated average molecular weight. The measurement is carried out by means of a number of high-speed devices for GPC (Toso company, HLC-8120GPC, the detection method using a differential refractive index) with columns TSKguard Super H-L (inner diameter 4.6 mm x 35 mm), TSKgel Super H4000 (inner diameter 6 mm x 150 mm) x 2 and TSKgel SuperH2000 (inner diameter 6 mm x 150 mm) (all columns manufactured by Toso) and THF as mobile phase with flow rate of 0.6 ml/min

If the interaction between polymer of lactic and glycolic acids and physiologically active substance is mariannae interaction, the interaction takes place is between the physiologically active substance and the terminal carboxylic acid polymer of lactic and glycolic acids. When low molecular weight fraction is present in a large number of physiologically active substance easily interacts with a low molecular weight polymer of lactic and glycolic acid having a high reactivity. Physiologically active substance, part of the injection means prolonged action, which is released in the process of receiving and at the initial stage of action, is mainly physiologically active substance interacting with a low molecular weight fraction of the polymer of lactic and glycolic acids. To increase the number of physiologically active substances and to reduce its initial release, it is necessary to reduce below a certain level, the content of low molecular weight fractions of the specified polymer of lactic and glycolic acids and the ratio of the average molecular weight and srednekamennogo molecular weight. For example, to obtain a preparation with prolonged action, releasing the drug for one month, it is desirable to use a polymer of lactic and glycolic acid having an average molecular weight of from about 8,000 to about 15,000, the ratio of the average molecular weight and srednekamennogo molecular weight of about 1,90 or less, preferably from about 1.40 to about 1,90, more preferably from the eye is about to 1.45 to about 1.80, and low molecular weight fraction with an average molecular weight of 3000 or less in an amount of about 9% or less, preferably from about 3% to about 9%.

"Polymer of lactic and glycolic acids can be obtained dehydrating polycondensation without the catalyst of lactic acid and glycolic acid (patent application Japan 61-28521) or by polymerization with ring opening of the lactide and the cyclic complex diapir, such as glycolide and the like (Encyclopedic Handbook of Biomaterials and Bioengineering Part A: Materials, Volume 2, Marcel Dekker, Inc., 1995).

The polymer of lactic and glycolic acids, obtained by dehydrating polycondensation without the catalyst of lactic acid and glycolic acid usually contains a large amount of low molecular weight fraction and the ratio of the average molecular weight and srednekamennogo molecular weight of about 2 or more. The average molecular weight of the polymer of lactic and glycolic acids according to the present invention is from about 5,000 to about 15,000. The content of the low molecular weight fraction with a molecular weight of about 3,000 or less may vary depending on the average molecular weight, and, when the average molecular weight equal to about 10,000, the amount of low molecular weight fraction with a molecular weight of about 3000 or less is about 10% or more.

The resulting polymer of lactic and glycolic acid which t can be cleaned using an organic solvent, receiving the final polymer.

Examples of the organic solvent used in the present invention, preferably include hydrophilic or soluble in water, organic solvents, such as, for example, acetone, tetrahydrofuran, dioxane, dimethylformamide and dimethylsulfoxide, and for this purpose it is preferable to use acetone.

The amount of added water and an organic solvent has no particular restrictions. However, when the amount of water is too large, it is impossible to sufficiently reduce the content of low molecular weight fractions, and it is therefore difficult to obtain a final polymer. On the other hand, when the amount of water is insufficient, the polymer bad precipitates, therefore worsening the output, and the formed polymer with a higher molecular weight compared with the desired molecular weight. The amount of water to 100 parts of an organic solvent is generally about 5-50 parts, preferably from about 10 to about 45 parts, more preferably from about 24 to about 40 parts, especially preferably about 40 parts. For example, 10 g of the polymer of lactic and glycolic acid are dissolved in 100 ml of acetone, an organic solvent, gradually add 40 ml of purified water, stirring acceptable method for the deposition con is knogo polymer, which can be dried in a suitable way. If the final polymer cannot be obtained in the process of performing a single stage of dissolution and deposition, this procedure can be repeated.

The product of prolonged action of the present invention as the base is preferably used a polymer of lactic and glycolic acid with a ratio of the average molecular weight and srednekamennogo molecular weight of about 1,90 or less or its salt, or a polymer of lactic and glycolic acid with an average molecular weight from about 11600 to about 14000 or its salt. The molar ratio of lactic acid and glycolic acid polymer is preferably from 100/0 to 40/60. Physiologically active substance is preferably derived LH-RH and its derivative LH-RH is preferably a peptide expressed by the formula

5-oxo-Pro-His-Trp-Ser-Tyr-DLeu-Leu-Arg-Pro-NH-C2H5

or its acetate. Derived LH-RH or its salt is preferably present in an amount of from about 5% (wt./wt.) to about 24% (wt./wt.). In addition, the preferred product of prolonged action, does not contain gelatin and releasing the physiologically active substance or its salt, at least within two weeks.

The method of producing microcapsules

Thus obtained polymer of lactic and glycolic acids can the be used as the basis for the preparation of prolonged action. The following describes the method of obtaining the composition of prolonged action of the present invention, for example, microcapsules containing a physiologically active substance or its salt and a polymer of lactic and glycolic acid or its salt.

(I) a Method of drying in the water

(i) the production method of the emulsion oil-in-water

In accordance with the method of the present invention will first get the solution of polylactic acid and glycolic acid or its salts in an organic solvent. The organic solvent used for obtaining the product of prolonged action of the present invention, preferably has a boiling point equal to 120°With or below.

As the organic solvent can be used, for example, halogenated hydrocarbons (e.g. dichloromethane, chloroform, dichloroethane, trichloroethane, carbon tetrachloride and the like), ethers (e.g. ethyl ether, isopropyl ether, and the like), esters of fatty acids (e.g. ethyl acetate, butyl acetate and the like), aromatic hydrocarbons (e.g. benzene, toluene, xylene and the like), alcohols (e.g. ethanol, methanol and the like) and acetonitrile, and a mixture of these solvents. The preferred organic solvent for the polymer of lactic and glycolic acids or salts thereof I have is dichloromethane.

The concentration of polymer of lactic and glycolic acids in solution of an organic solvent may vary depending on the molecular weight of the polymer of lactic and glycolic acids and the type of organic solvent. For example, using dichloromethane as the organic solvent concentration is usually chosen from about 0.5 to about 70 wt.%, more preferably from about 1 to about 60 wt.%, particularly preferably from about 2 to about 50 wt.%.

Then add a physiologically active substance or its salt, which is dissolved or dispersed in the resulting solution of polymer of lactic and glycolic acid in an organic solvent. The resulting solution in an organic solvent containing the composition of the physiologically active substance or its salt and a polymer of lactic and glycolic acid or its salts, is added to the aqueous phase to form an emulsion of the oil phase in the aqueous phase, after which the solvent in the oil phase is evaporated or diffused into the aqueous phase with the formation of microcapsules. The volume of the aqueous phase is usually chosen so that it was about 1-10000 times, more preferably about 5-50000 times, most preferably approximately 10-2000 times the volume of oil phase. In addition to the above components to the aqueous phase may be added emulsifier. You can use uboy emulsifier, allows to obtain a stable emulsion of oil phase in the aqueous phase. For example, it is possible to use anionic surfactants (sodium oleate, sodium stearate, sodium lauryl sulfate and the like), nonionic surfactants (esters of fatty acids of polyoxyethylenesorbitan) (tween 80, tween 60, Atlas Powder), polyoxyethylene derivative of castor oil (HCO-60, HCO-50 firm Nikko Chemical), polyvinylpyrrolidone, polyvinyl alcohol, carboxymethylcellulose, lecithin, gelatin, and hyaluronic acid. These substances can be used alone or in combination with each other. The concentration of these substances is preferably in the range from about 0.0001 to 10 wt.%, more preferably from about 0.001 to 5 wt.%

In addition to the above components to the aqueous phase may be added a substance regulating the osmotic pressure. You can use any regulator of osmotic pressure, is able to create the desired osmotic pressure when introduced into the aqueous solution.

Examples of regulators of osmotic pressure include polyhydric alcohols, monohydroxy alcohols, monosaccharides, disaccharides, oligosaccharides and amino acids or their derivatives.

As polyhydric alcohols can be used, for example, trivalent alcohols such as glycerin and the like, acetone alcohols, such as Arabic, xylitol, adonit and the like, and setiathome alcohols, such as mannitol, sorbitol, dulcet and the like. The preferred alcohols are setiathome alcohols, in particular mannitol.

Examples of monohydroxy alcohols include methanol, ethanol and isopropyl alcohol, and ethanol is preferred.

As monosaccharides can be used, for example, pentoses, such as arabinose, xylose, ribose, 2-deoxyribose, and the like, and hexose, such as glucose, fructose, galactose, mannose, sorbose, rhamnose, fucose, and the like, and hexose are more preferred.

As oligosaccharides can be used, for example, trisaccharide, such as maltotriose, raffinose and the like, and tetrasaccharide, such as stachyose and the like, and trisaccharide are more preferred.

As derivatives of monosaccharides, disaccharides and oligosaccharides can be used, for example, glucosamine, galactosamine, glucuronic acid and galacturonic acid.

As amino acids, you can use any L-amino acids, examples of which include glycine, leucine and arginine. Among these amino acids, more preferred is L-arginine.

These regulators of osmotic pressure can be used separately or combined in the and with each other.

Regulators of osmotic pressure is used in such a concentration that the osmotic pressure of the external water phase was approximately 1/50 - 5 times, preferably about 1/25 - 3 times the osmotic pressure of saline. When using mannitol as a regulator of osmotic pressure, its concentration is preferably 0.5 to 1.5%.

For removal of the organic solvent can be used or a similar way. Examples of such methods include evaporation of the organic solvent under normal pressure or under gradually decreasing pressure and stirring propeller stirrer, a magnetic stirrer or by means of ultrasonic devices; evaporation of the organic solvent in regulating the degree of vacuum using a rotary evaporator and the gradual removal of organic solvent by using a membrane for dialysis.

Thus obtained microcapsules centrifuged or filtered to remove free physiologically active substance or its salt, after which the substance that keeps the drug, and the emulsifier on the surface of the microcapsules are washed several times with distilled water and liofilizovannye microcapsules again dispersed in distilled water.

As the microcapsule truly is obreteniyu as the basis used is a polymer of lactic and glycolic acid with a ratio of the average molecular weight and srednekamennogo molecular weight, equal 1,90 or less, or its salt, or a polymer of lactic and glycolic acid with an average molecular weight from about 11600 to about 14000 or its salt, such microcapsule may contain the drug in large quantities, and therefore, upon receipt of the microcapsules is not necessary to use a substance that keeps the drug, such as gelatin and thickener.

These polymers can preferably be used to obtain compositions with prolonged action, releasing the drug at least for two weeks.

At the stage of obtaining microcapsules may be added a substance that prevents aggregation of particles. As substances that prevent aggregation, can be used, for example, water-soluble polysaccharides such as mannitol, lactose, glucose and starches (e.g. corn starch and the like), an amino acid, such as glycine, and protein such as fibrin and collagen. Particularly suitable is mannitol.

Water and organic solvent remaining in the microcapsule after drying, if necessary, can be removed by heating the microcapsule in conditions that do not cause its destruction. The heating is preferably carried out at a temperature that is equal to or slightly higher than the intermediate glass transition temperature of the microcapsules, the mouth of blennow using a differential scanning calorimeter, increasing the temperature speeds of 10-20°in a minute. More preferably, the microcapsules are heated at a temperature equal to the intermediate glass transition temperature of the microcapsules, or in the temperature interval from the intermediate glass transition temperature of the microcapsules to a temperature that is about 30°above the intermediate glass transition temperature. The microcapsules preferably heated in the temperature range from the intermediate glass transition temperature of the microcapsules to a temperature which is 10°above the intermediate glass transition temperature, more preferably in the temperature range from the intermediate glass transition temperature to a temperature which is 5°above the intermediate glass transition temperature.

Heating time may vary depending on the number of microcapsules and usually ranges from about 12 hours to 168 hours, preferably from about 24 hours to 120 hours, particularly preferably from about 48 hours to 96 hours after the microcapsule predetermined temperature.

The method of heating does not have specific restrictions, if achieved, uniform heating of all of the microcapsules.

Heating and drying can be performed, for example, in a thermostatic chamber, the chamber fluidized bed, a movable chamber, a drying oven or a microwave oven. Preference is sustained fashion way is the heating and drying in a thermostatic chamber.

(ii) the production method of the emulsion water in oil in water (W/O/W)

First get the solution of polylactic acid and glycolic acid or its salts in an organic solvent, and the thus obtained solution in an organic solvent is an oil phase. The method of obtaining the oil phase is similar to the method described in the above section (I) (i). The concentration of polymer of lactic and glycolic acid in an organic solvent may vary depending on the molecular weight of the polymer of lactic and glycolic acids and the type of organic solvent. For example, using dichloromethane as the organic solvent concentration is usually chosen in the range of from about 0.5 to about 70 wt.%, more preferably from about 1 to about 60 wt.%, particularly preferably from about 2 to about 50 wt.%.

Then you get the solution or dispersion of a physiologically active substance or salt [solvent is water or a mixture of water and alcohols (e.g. methanol, ethanol and the like)].

The concentration of added physiologically active substance or its salt is usually from 0.001 mg/ml to 10 g/ml, preferably from 0.1 mg/ml to 5 g/ml, more preferably from 10 mg/ml to 3 g/ml.

If the above-described physiologically active substance has a basic group such as amino group, salts of the physical and logicheskie active substances include salts with inorganic acids (also referred to as free inorganic acids (for example, carbonic acid, carbonate acid, hydrochloric acid, sulfuric acid, nitric acid, boric acid, and the like) and organic acids (also referred to as free organic acids (e.g. succinic acid, acetic acid, propionic acid, triperoxonane acid and the like).

If the physiologically active substance has an acidic group such as carboxyl group, salts of physiologically active substances include salts with inorganic bases (also referred to as free inorganic bases) (for example, alkali metals such as sodium and potassium, alkaline earth metals such as calcium, magnesium and the like) and organic bases (also referred to as free organic basis) (for example, organic amines such as triethylamine, basic amino acids such as arginine and the like). In addition, physiologically active peptides can form complex compounds of metals (e.g., copper complex, a complex of zinc, and the like). When the physiologically active substance is derived LH-RH, preferably add acetic acid.

As a solubilizer and stabilizer can be used known compounds. To dissolving or dispersing a physiologically active substances the creation or additive, the mixture is heated, shaken and stirred so as to preserve the activity of the specified substances, thus obtaining an aqueous solution, representing the internal aqueous phase.

Thus obtained internal aqueous phase and an oil phase emuleret by known methods such as homogenization and ultrasound treatment, emulsion water in oil.

The volume of mixed oil phase is about 1 to 1,000 times, preferably 2-100 times, more preferably about 3-10 times the volume of the internal aqueous phase.

The viscosity of the resulting emulsion water in oil is usually equal to about 10-10000 CP, preferably about 100-5000 CP, more preferably about 500-2000 the centipoise at a temperature of about 12-25°C.

Then, the resulting emulsion water in oil, containing a physiologically active substance or its salt and a polymer of lactic and glycolic acid or its salt, is added to the aqueous phase with the formation of emulsions of the water (W) (internal water phase/oil phase (W/O) in the external aqueous phase (W), after which the solvent in the oil phase is evaporated or diffused into the external aqueous phase with the formation of microcapsules. The volume of external aqueous phase is usually chosen so that it was about 1-10000 times, more preferably about 5-50000 times, most preferably approximately 10-2000 times the volume of the mA oil phase.

In addition to the above components to the aqueous phase can be added emulsifier and regulator of osmotic pressure, and the methods of making microcapsules are similar to those described in the above section (I)(i).

(II) the Method of separation of the phases

Upon receipt of the microcapsules by the method of the present invention to the solution of the physiologically active substance or its salt and a polymer of lactic and glycolic acid in an organic solvent described in the method of drying in the water section (I), with stirring, gradually add Coatesville agent for deposition and curing of the microcapsules. Volume laservideo agent may be about 0.01 to 1000 times, preferably about 0.05 to 500 times, especially preferably about 0.1 to 200 times the volume of oil phase.

Type laservideo agent has no specific limitation, if it is a polymer compound mineral or vegetable oil that is compatible with an organic solvent not dissolving the complex of the physiologically active substance or salt and a polymer of lactic and glycolic acids or salts thereof. In particular, you can use silicone oil, sesame oil, soybean oil, corn oil, cottonseed oil, coconut oil, linseed oil, mineral oil, n-hexane or n-heptane. You can use a mixture of two or more is the number of specified substances.

Thus obtained microcapsules exhale, washed several times with heptane or similar substance, to remove Coatesville agent from the composition comprising a physiologically active substance or its salt and a polymer of lactic and glycolic acid or its salt, and dried under reduced pressure. Microcapsules alternative washed by the method described in the method of drying water liofilizovannyh microcapsules in the above section (I) (i), after which the microcapsules are heated and dried.

(III) Spray drying

Upon receipt of the microcapsules by the method according to the present invention a solution of a physiologically active substance or its salt and a polymer of lactic and glycolic acid or its salts in an organic solvent described in the method of drying in the water section (I), is sprayed into the drying chamber of the spray drying using a spray nozzle, after which the organic solvent in finely fragmented droplets evaporate in a short time with the formation of microcapsules. Examples of the spray nozzle includes a nozzle for spraying in a fluidized bed, a nozzle for spraying under pressure, a nozzle with a rotary drive. Then, if necessary, the microcapsules are washed, lyophilized, again washed and dried the same way as the drying in water described in (I).

When receiving dosiro the authorized dosage forms, non of the above microcapsule, a solution of a physiologically active substance or its salt and a polymer of lactic and glycolic acid or its salts in an organic solvent, which is described in the method of producing microcapsules by drying in the water section (I), dried, vaporizing the organic solvent and water by regulating the degree of vacuum using, for example, a rotary evaporator, after which the resulting product is pulverized in a jet mill, thus obtaining fine powder (hereinafter also referred to as microparticles).

Finely ground powder is washed in a manner analogous to the method of drying in the water used in the preparation of the microcapsules (I), lyophilized, heated and dried.

The obtained microcapsules or fine powder can release the drug at a lower flow rate of the used polymer of lactic and glycolic acids.

The composition of the prolonged action of the present invention may have any shape, such as a microsphere, microcapsule, or fine powder (particles), and the microcapsule is the most acceptable form.

The composition of the prolonged action of the present invention can be used without further processing or as feedstock for different dosage forms for intravenous and subcutaneous is doing as injectable or implantable means, and for insertion inside the body through the mucous membrane and oral (for example, in the form of hard capsules, soft capsules, and the like) and use in the form of solid preparations such as granules, powder and the like, or liquids, such as syrup, emulsion, suspension and the like for insertion into the nose, rectum or uterus.

For example, for the manufacture of compositions prolonged action of the present invention in the form of injectable contraceptives receive aqueous suspension together with a dispersant such as a surfactant, such as tween-80, HCO-60 and the like, and polysaccharides such as sodium hyaluronate, carboxymethylcellulose, Argent sodium and the like), a preservative (such as methylparaben and propylparaben), a substance for isotonic solution (such as sodium chloride, mannitol, sorbitol, glucose and Proline)or a specified composition is dispersed in an oil suspension together with a vegetable oil, such as sesame or corn oil, while receiving the injection tool prolonged action, which can be used without further processing.

The diameter of the particles of the composition of the prolonged action of the present invention, designed for use in suspension injectable contraceptives, should sootvetstvovat the desired degree of dispersion and permeability of the needle. For example, the average particle diameter may be equal to about 0.1-300 microns, preferably about 0.5 to 150 μm, more preferably about 1-100 μm.

The composition of the prolonged action of the present invention in the form of aseptic preparation may be obtained by methods that include, but are not limited to, aseptic execution of all stages of the process of obtaining, sterilization with gamma rays, adding an antiseptic and the like.

Since the composition of the prolonged action of the present invention has low toxicity, it can be used as a safe drug to a mammal (for example, man, cattle, pigs, dogs, cats, mice, rats, rabbits, and other mammals), while the dose of the composition of the prolonged action of the present invention may vary depending on the type and content of the physiologically active substance, the type of dosage form, duration of release of the physiologically active substance subject to the treatment of disease, animal species and an effective amount of physiologically active substances. Single dose of the physiologically active substance may preferably be for an adult from about 0.01 mg to 10 mg/kg body weight, more predpochtite the flax from about 0.05 mg to 5 mg/kg of body weight, for example, when used in the preparation of long action with the release of the active substance within six months.

Single dose compositions prolonged action may preferably be for an adult from about 0.05 mg to 50 mg/kg body weight, more preferably from about 0.1 mg to 30 mg/kg of body weight.

The introduction can be selected depending on the type and content of the physiologically active substance is used as the basis, dosage form, duration of release of the physiologically active substance for treatment of the disease and the type of animal, for example, the drug can be administered once every few weeks once a month once every few months (for example, three months, four months, six months and so on and so on

The composition of the prolonged action of the present invention can be used as a means to prevent or treat various diseases depending on the type of physiologically active substance contained in the composition; for example, when the physiologically active substance is derived LH-RH, such a composition can be used to prevent or treat hormonal diseases, in particular cancer tumors that are dependent on the sex of the mountains is it (for example, prostate cancer, uterine cancer, breast cancer, pituitary tumor, etc.), diseases dependent on sex hormone, such as prostatomegaly, endometriosis, uterine fibroids, premature puberty, dysmenorrhea, amenorrhea, premenstrual syndrome, multi-chambered ovary, and the like, to prevent recurrence of breast cancer after the operation for premenopausal breast cancer, for the prevention and treatment of Alzheimer's disease or autoimmune diseases, and contraception (or the prevention or treatment of infertility in order to restore activity after discontinuation of contraceptives). In addition, the above composition can also be used as a means for prevention or treatment of benign or malignant tumors that are not caused by sex hormone, but is sensitive to LH-RH.

Thus, the introduction to a mammal an effective dose of a therapeutic or prophylactic agent of the present invention makes possible the prevention or treatment of hormonal diseases, in particular cancer tumors that are dependent on sex hormone (e.g., prostate cancer, uterine cancer, breast cancer, pituitary tumors, and the like), diseases dependent on sex hormone, such as about cytomegalia, endometriosis, uterine fibroids, premature puberty, dysmenorrhea, amenorrhea, premenstrual syndrome, multi-chambered ovary, and the like, prevention of conception and prevention of relapse of breast cancer after the operation for premenopausal breast cancer.

Examples

The present invention will be further explained in more detail by means of examples, comparative examples and experimental examples, but the present invention is not limited to these examples.

Example A1

10 g of the polymer of lactic and glycolic acid with an average molecular weight of 9700 and srednekamennogo molecular weight 5030 synthesized by dehydrating polycondensation of lactic acid and glycolic acid, are dissolved in 100 ml of acetone and with stirring, added dropwise 40 ml of purified water to precipitate the polymer. The solution also besieged syrupy polymer in the form of sticky starch decanted and the polymer is dried in vacuum. The polymer yield after drying is of 8.37 g, average molecular weight equal to 10500 and srednekislye molecular weight equal to 6700.

Example A2

to 4.87 g of the polymer obtained in example A1 are dissolved in 8,03 g of dichloromethane in the oil phase. The oil phase is mixed with an aqueous phase, representing the solution 0,597 g of the acetate of peptide And 0.6 ml of cleansing the Noi water, and emuleret with a speed of 25,000 rpm using Polytron, while receiving the emulsion water in oil. Emulsion water in oil is added to 1000 ml of 0.1% aqueous solution of polyvinyl alcohol at 15°and to emulsify water in oil in water (W/O/W) in a homogenizing mixer, rotating at 7000 revolutions/min Microcapsules utverjdayut, desalvatore them using a propeller stirrer for three hours, after which the microcapsules passing through a sieve with a mesh size of 200 mesh, separated and lyophilized after addition of 0.48 g of mannitol. Output liofilizovannyh microcapsules equal to 3.92 g, and the content of the peptide And is 10,18%.

Comparative example A1

Microcapsules obtained using a polymer of lactic and glycolic acids according to example A1 by way of example A2 with the release of 3.97 g, and the content of the peptide And of 9.50%.

Experimental example A1

The microcapsules obtained in example A2 and comparative example A1, dispersed in 0.3 ml of a dispersing medium (distilled water, which dissolved 0.25 mg of carboxymethylcellulose, 0.5 mg of Polysorbate 80 and 25 mg of mannitol) in the amount of 2.25 mg, calculated on the peptide and injected subcutaneously into the back of the male SD rats aged 7 weeks with a needle for injection 22G. At a predetermined time after the introduction of rats, kill, delete microcapsules remaining in the rst place is to be placed, and determine the content of peptide A, the value of which is divided by the initial number, thereby determining the percentage of the remaining amount, as shown in table 1. In addition, this table shows the correlation between the average molecular weight and srednekamennogo molecular weight (Mw/Mn) of the polymers used in example A2 and comparative example A1.

Table 1
Comparative example A1Example A2
The ratio Mw/Mn1,93of 1.57
1 day84,64%91,17%
2 weeks32,2%54,31%
4 weeks2,54%10,28%

As shown in table 1, when used for the production of microcapsules of polymer in example A2 with a ratio of Mw/Mn equal to 1,90 or less achieved in the processing of acetone, suppressed initial release of the peptide And of the microcapsules and provides prolonged release over a long period of time equal to four weeks.

Example A3

185,7 g of the polymer of lactic and glycolic acid with an average molecular weight 10600 and srednekamennogo molecular weight of 6600 dissolve in to 30.1 g of dichloromethane, bringing the temperature up to 29.5°C. Give 330,2 g of the obtained solution in an organic solvent, is mixed with an aqueous solution obtained by dissolving 15.62 wide g of the acetate of peptide And 15,31 g of distilled water, previously heated to 54,3°C, stirred for 1 minute and get a crude emulsion, which is then emuleret for two minutes in a homogenizer with a rotation speed of 10,000 revolutions per minute, emulsion water in oil (W/O). The resulting emulsion water in oil is cooled to 17.8°C, poured into 25 liters of 0.1% (wt./wt.) an aqueous solution of polyvinyl alcohol (EG-40, firms Nihongoseikagaku), which is pre-adjusted to 17.9°C for 1 minute and 16 seconds, and stirred with a speed 7005 rpm in the device HOMOMIC LINE FLOW (firm Tokushukika)to give the emulsion water in oil in water (W/O/W). The resulting emulsion water in oil in water is stirred for 3 hours to evaporate dichloromethane or dichloromethane diffusion into the external aqueous phase, an oil phase utverjdayut and filtered through a sieve with openings of 75 μm, to form microcapsules with continuous rotation in a centrifuge (H-600S, firms Kokusanenshinki) with a speed of 2000 rpm and harvested. The collected microcapsules again dispersed in a small amount of distilled water, filtered through a sieve with apertures 90 μm, dissolved, adding 17,2 g man the ITA and lyophilized, receiving the powder. The output of the microcapsules is equal to 76.4%, and the content of the peptide in the microcapsules is 8,79%.

Experimental example A2

Approximately 26 mg of the microcapsules described in example A3, dispersed in 0.3 ml of a dispersing medium (distilled water, which dissolved 0.15 mg of carboxymethylcellulose, 0.3 mg of Polysorbate 80 and 15 mg of mannitol) and injected subcutaneously into the back of the male SD rats aged 7 weeks with a needle for injection 22G. At a predetermined time after the introduction of rats, kill, delete, microcapsules, remaining at the injection site, and determine the content of peptide A, the value of which is divided by the initial number, thereby determining the percentage of the remaining amount, as shown in table 2.

Table 2
Time1 day1 week2 weeks3 weeks4 weeks5 weeks
The remaining number90,29%68,06%36,63%was 12.75%4,48%1,12%

As shown in table 2, despite the high content of the physiologically active substance in the microcapsules described in example A3, the number of physiologically active substances, the remaining is to be one day after injection, is 90%. Thus, when the ratio Mw/Mn of the polymer does not exceed 1.6, achieved a significant suppression of the high initial release of physiologically active substances. In addition, these microcapsules provide a release of a physiologically active substance at a constant speed for a long period of time.

Example B1

197,7 g of the polymer of lactic and glycolic acid with an average molecular weight of 12600 and srednekamennogo molecular weight of 6400 dissolved in 320,1 g of dichloromethane, filtered through a filter press with a hole size of 0.2 μm (EMFLOW, DFA4201FRP) and bring the temperature to about 30,0°C. Give 330,1 g of the solution, is mixed with an aqueous solution obtained by dissolving 15,68 g of the acetate of peptide And 15,31 g of distilled water, previously heated to 56,0°C, stirred for 1 minute and get a crude emulsion, which emuleret within two minutes with a rotation speed of 10,000 rpm with emulsion water in oil. The resulting emulsion water in oil is cooled to 18.2°C, poured into 25 liters of 0.1% (wt./wt.) solution of polyvinyl alcohol (EG-40, firms Nihongoseikagaku), which is pre-adjusted to 18.4°C for 1 minute and 46 seconds, and stirred with a speed 7007 rpm using a device HOMOMIC LINE FLOW (firm Tokushukika)to give the emulsion in the and in the oil-in-water (W/O/W). The resulting emulsion water in oil in water is stirred for 3 hours to evaporate dichloromethane or dichloromethane diffusion into the external aqueous phase, then the oil phase utverjdayut and filtered through a sieve with openings of 75 μm, to form microcapsules with continuous rotation in a centrifuge (H-600S, firms Kokusanenshinki) with a speed of 2000 rpm and harvested. The collected microcapsules again dispersed in a small amount of distilled water, filtered through a sieve with apertures 90 μm, dissolved, adding and 17.2 g of mannitol and lyophilized, receiving the powder. The output of the microcapsules equal 73,47%, and the content of the peptide in the microcapsules is 8.43 per cent.

Experimental example B1

About 26,7 mg of microcapsules described in example B1, is dispersed in 0.3 ml of a dispersing medium (distilled water, which dissolved 0.15 mg of carboxymethylcellulose, 0.3 mg of Polysorbate 80 and 15 mg of mannitol) and injected subcutaneously into the back of the male SD rats aged 7 weeks with a needle for injection 22G. At a predetermined time after the introduction of rats, kill, delete, microcapsules, remaining at the injection site, and determine the content of peptide A, the value of which is divided by the initial number, thereby determining the percentage of the remaining amount, as shown in table 3.

Table 3
Time1 day1 week2 weeks3 weeks4 weeks5 weeks
The remaining number82,43%68,33%47,07%23,58%9,05%2,08%

As shown in table 3, the microcapsules described in example B1, can contain physiologically active substance in a large amount even without the use of gelatin and is able to significantly suppress the initial release of physiologically active substances, providing release of physiologically active substance over a long period of time.

Experimental example B2

About to 44.6 mg of microcapsules described in example B1, is dispersed in 1.0 ml of a dispersing medium (distilled water, which dissolved 0.15 mg of carboxymethylcellulose, 0.3 mg of Polysorbate 80 and 15 mg of mannitol) and injected subcutaneously in the back of the short-legged hounds weighing 7-12 kg with a needle for injection 22G. At a predetermined time after the introduction of dogs from Vienna front paws take a blood sample and determine the concentration of peptide a and testosterone; the results are shown in table 4.

Table 4
Time1 day1 week2 weeks3 weeks4 weeks5 weeks
Peptide Andof 2.210,3980,5250,4330,6030,358
Testosterone2,790,570,350,350,300,39

As shown in table 4, the microcapsules described in example B1, release of physiologically active substance over a long period of time and remain in the blood the necessary concentration of the physiologically active substance. In addition, the physiologically active substance is released into the blood, does not lose activity, due to which the drug remains effective.

Industrial applicability

Drug prolonged action of the present invention, including as the basis of a PLGA copolymer with a ratio of the average molecular weight and srednekamennogo molecular weight of about 1,90 or less or a polymer of lactic and glycolic acid with an average molecular weight from about 11600 to about 14000 or its salt, contains physiologically active substance in a large amount even without the use of gelatin and suppresses high initial release of fisiologicas and active substances, providing a stable rate of release for about one month.

Thus, the preparation according to this invention has such useful properties that can be obtained while reducing the manufacturing process and reduce costs, so as not to have to use a substance that keeps the drug, such as gelatin, and a thickener, which reduces the quantity of additives, making this product may contain the drug in a high concentration without the use of substances that hold the drug, and thickener; in addition, due to the increase of glass transition temperature can be obtained composition with prolonged action, releasing the drug at least for two weeks, and the drug high resistance.

1. The slow release composition containing a polymer of lactic and glycolic acid with a ratio of the average molecular weight and srednekamennogo molecular weight of about 1,90 or less or its salt and the peptide expressed by the formula

5-oxo-Pro-His-Trp-Ser-Tyr-Y-Leu-Arg-Pro-Z,

where Y represents DLeu, DAla, DTrp, DSer (tBu), D2Nal or DHis (ImBzl), and Z denotes HN-C2H5or Gly-NH2or its salt.

2. The slow release composition according to claim 1, in which the polylactic and lilleboe acid has an average molecular weight of from about 3000 to about 100,000.

3. The slow release composition according to claim 2, in which the polymer of lactic and glycolic acid has an average molecular weight of from about 8,000 to about 15,000.

4. The slow release composition according to claim 1, in which the low molecular weight fraction of the polymer of lactic and glycolic acids with a molecular weight of about 3000 or less is about 9% or less.

5. The slow release composition according to claim 4, in which the low molecular weight fraction of the polymer of lactic and glycolic acids with a molecular weight of about 3000 or less is from about 3% to about 9%.

6. The slow release composition according to claim 1, in which the specified polymer is characterized by a molar ratio of lactic acid and glycolic acid from 100:0 to 40:60.

7. The slow release composition according to claim 1, in which the specified polymer is characterized by a molar ratio of lactic acid and glycolic acid 70:30 to 80:20.

8. The slow release composition according to claim 1, in which the peptide is expressed by the formula

5-oxo-Pro-His-Trp-Ser-Tyr-DLeu-Leu-Arg-Pro-NH-C2H5,

or its acetate.

9. The slow release composition according to claim 1, in which the peptide or its salt is contained in an amount of from about 5% (wt./wt.) to about 24% (wt./wt.).

10. The slow release composition according to claim 1, which is designed for injection.

11. To the position of the slow release according to claim 1, which releases the peptide or its salt, at least within two weeks.

12. The slow release composition according to claim 1, which does not include a substance that keeps the drug.

13. The slow release composition according to claim 1, which contains no gelatine.

14. The method of obtaining a slow release composition according to claim 1, which comprises removing the solvent from the solution containing the peptide of the formula

5-oxo-Pro-His-Trp-Ser-Tyr-Y-Leu-Arg-Pro-Z,

where Y represents DLeu, DAla, DTrp, DSer (tBu), D2Nal or DHis (ImBzl), and Z denotes HN-C2H5or Gly-NH2or its salt and a polymer of lactic and glycolic acid with a ratio of the average molecular weight and srednekamennogo molecular weight of about 1,90 or less, or its salt.

15. The method according to 14, which comprises mixing and dispersing the peptide or its salt in the mixed solution of an organic solvent containing the polymer of lactic and glycolic acid with a ratio of the average molecular weight and srednekamennogo molecular weight of about 1,90 or less or its salts and the subsequent removal of the organic solvent.

16. The method according to item 15, in which the peptide or its salt used in the aqueous solution containing the peptide or its salt.

17. The pharmaceutical preparation containing the slow release composition according to claim 1 with dozirovki mg/kg body weight.

18. Medicinal product for preventing or treating prostate cancer, prostatomegaly, endometriosis, fibroids, metrofibroma, premature puberty and dysmenorrhea or for contraception that contains a slow release composition according to claim 1.

19. Drug to prevent recurrence of breast cancer after the operation for premenopausal breast cancer, which contains a slow release composition according to claim 1.

20. A method of preventing or treating prostate cancer, prostatomegaly, endometriosis, fibroids, metrofibroma, premature puberty and dysmenorrhea, or a contraceptive, which comprises the administration to a mammal an effective dose of slow release composition according to claim 1.

21. The way to prevent recurrence of breast cancer after the operation for premenopausal breast cancer, which comprises the administration to a mammal an effective dose of slow release composition according to claim 1.

22. A method of obtaining a polymer of lactic and glycolic acid having an average molecular weight of from about 8,000 to about 15,000 and the ratio of the average molecular weight and srednekamennogo molecular weight of about 1,90 or less, or its salt, which comprises adding to the water to organic solvent, containing a polymer of lactic and glycolic acid with an average molecular weight from about 5,000 to about 15,000, with a ratio of less than about 5-50 parts (volumes) per 100 parts of organic solvent.

23. A method of producing a polymer according to item 22, in which the organic solvent is hydrophilic.

24. A method of producing a polymer according to item 23, in which the hydrophilic organic solvent is acetone.

25. A method of producing a polymer according to item 22, in which the ratio of water to 100 parts of organic solvent is from about 10 to about 45 parts (volumes).

26. A method of producing a polymer according to item 22, in which the ratio of water to 100 parts of organic solvent is about 40 parts (volumes).

27. The polymer of lactic and glycolic acid having an average molecular weight of from about 8,000 to about 15,000 and the ratio of the average molecular weight and srednekamennogo molecular weight of about 1,90 or less, or salt and having a molecular ratio of lactic acid and glycolic acid from 100:0 to 40:60, which is produced by adding water to the organic solvent containing the polymer of lactic and glycolic acid with an average molecular weight from about 5,000 to about 15,000, with a ratio of less than about 5-50 parts (volumes) per 100 parts organizes the first solvent.

28. The use of the polymer of lactic and glycolic acids in item 27 or its salt to obtain a slow release composition that does not contain gelatin.

29. The microsphere containing polymer of lactic and glycolic acid with an average molecular weight from about 11600 to about 14000 or its salt and a peptide of the formula

5-oxo-Pro-His-Trp-Ser-Tyr-Y-Leu-Arg-Pro-Z,

where Y represents DLeu, DAla, DTrp, DSer(tBu), D2Nal or DHis (ImBzl), and Z denotes HN-C2H5or Gly-NH2or its salt, and do not contain gelatin.

30. Microsphere according to clause 29, in which the peptide or its salt is a peptide of the formula

5-oxo-Pro-His-Trp-Ser-Tyr-DLeu-Leu-Arg-Pro-NH-C2H5,

or its acetate.

31. Microsphere according to clause 29, which contains the peptide or its salt in an amount of from about 5% (wt./wt.) to about 24% (wt./wt.).

32. Microsphere according to clause 29, which represents a microcapsule.

33. Microsphere according to clause 29, which releases the peptide or its salt, at least for more than 2 weeks.

34. Medicinal product for preventing or treating prostate cancer, prostatomegaly, endometriosis, fibroids, metrofibroma, premature puberty and dysmenorrhea or for contraception that contains a microsphere according to clause 29.

35. Drug to prevent recurrence of breast cancer after the operation for preclimacteric the CSOs breast cancer, which contains a microsphere according to clause 29.

36. A method of preventing or treating prostate cancer, prostatomegaly, endometriosis, fibroids, metrofibroma, premature puberty and dysmenorrhea, or a contraceptive, which comprises the administration to a mammal an effective dose of the microspheres in clause 29.

37. The way to prevent recurrence of breast cancer after the operation for premenopausal breast cancer, which comprises the administration to a mammal an effective dose of the microspheres in clause 29.

Priority items:

29.06.2001 according to claims 1-18, 22-27;

06.11.2001 on PP-21, 28-37.



 

Same patents:

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to novel derivatives of benzimidazole of the general formula (I): wherein A represents -CH2- or -C(O)-; Y represents -S- or -NH-; R1 and R2 represent independently hydrogen atom, (C1-C8)-alkyl, (C5-C9)-bicycloalkyl optionally substituted with one or some similar or different (C1-C6)-alkyl radicals, or radical of the formula -(CH2)n-X wherein X represents amino-group, (C3-C7)-cycloalkyl and other values of radicals also given in the invention claim; R3 represents -(CH2)p-W-(CH2)p'-Z3 wherein W3 represents a covalent bond, -CH(O)- or -C(O)-; Z3 represents (C1-C6)-alkyl, aryl radical, heteroaryl and other values of radical also; V3 represents -O-, -S-, -C(O)-, -C(O)-O-, -SO2- or a covalent bond; Y3 represents (C1-C6)-alkyl radical optionally substituted with one or some halogen-radicals, amino-group, di-((C1-C6)-alkyl)-amino-group, phenylcarbonylmethyl, heterocycloalkyl or aryl radicals; p, p' and p'' represent independently a whole number from 0 to 4; R4 represents radical of the formula: -(CH2)s-R''4 wherein R''4 represents heterocycle comprising at least one nitrogen atom and optionally substituted with (C1-C6)-alkyl or aralkyl, and other values of radicals given in the invention claim also. Also, invention relates to a pharmaceutical composition showing antagonistic property with respect to GnRH and based on these compounds. Also, using above proposed compounds for preparing a medicament is considered. Invention provides synthesis of novel compounds, preparing pharmaceutical composition and medicament based on thereof in aims for treatment of such diseases as endometriosis, fibroma, polycystic ovary, breast, ovary and endometrium cancer, gonadotropic hypophysis desensitization in medicinal stimulation of ovary in fertility treatment in females.

EFFECT: valuable medicinal properties of compounds and pharmaceutical composition.

18 cl, 2 tbl, 538 ex

FIELD: medicine.

SUBSTANCE: invention relates to method for treatment, prevention and alleviation of testosterone deficit development; or symptoms associated with testosterone deficit in mammalian females. Claimed method includes application of gel containing (wt.%) testosterone 0.1-10; isopropylmyristate 0.1-5 %; ethanol 98 % or less; and thickening agent 0.1-5 % on skin of subject.

EFFECT: convenient method for maintenance of daily reproducible testosterone levels.

8 cl, 5 tbl, 4 ex

FIELD: pharmaceutical industry.

SUBSTANCE: pharmaceutical form contains (i) peptides with aggregation tendency in the form of their salts: acetates, gluconates, glucuronates, lactates, citrates, benzoates, or phosphates, which are dissolved of dispersed, and (ii) acids corresponding to above-listed salts.

EFFECT: lowered aggregation tendency and improved release of active substance resulting in improved bioavailability of peptide active substances.

22 cl, 9 tbl, 3 ex

FIELD: organic chemistry, pharmacy.

SUBSTANCE: invention relates to a new derivative of bicyclic heteroaromatic compound of the general formula (I) or its pharmaceutically acceptable salt eliciting agonistic activity with respect to luteinizing hormone (LH). Compounds can be used for preparing medicinal agents for control ability for conception. In compounds of the general formula (I) R1 represents R7 wherein R7 represents (C6-C10)-aryl optionally substituted with halogen atom at ortho- and/or meta-position; NHR8, OR8 wherein R8 means (C1-C8)-alkyl that can be substituted with halogen atom, (C1-C8)-alkylcarbonyl, (C1-C8)-alkylcarbonyloxy-group, phenyl, (C6-C10)-arylcarbonylamino-group, 5-methyl-2-phenylimidazol-4-yl, (C6)-heterocycloalkyl wherein 1-2 heteroatoms are taken among nitrogen and oxygen atoms, ethyloxycarbonylmethylthio-(C1-C4)-alkoxy-group, amino-group, (C6-C7)-heteroaryl; or (C5-C6)-heteroaryl comprising nitrogen, oxygen or sulfur atom as a heteroatom; R2 represents (C1-C8)-alkyl or (C6-C10)-aryl optionally substituted with one or more substitutes taken among (C1-C8)-alkoxy-group; or (C5-C6)-heteroaryl comprising nitrogen, oxygen or sulfur atom as a heteroatom; R3 represents (C1-C8)-alkyl possibly substituted with (C6-C14)-aryl possibly substituted with halogen atom, (C1-C4)-alkoxy-group, (C1-C4)-alkoxycarbonyl, mono- or tri-(C6-C10)-cycloalkyl, (C6-C10)-aryl, (C5-C6)-heteroaryl comprising nitrogen, oxygen or sulfur atom as a heteroatom; (C5-C7)-heterocycloalkyl comprising 2 heteroatoms taking among nitrogen or oxygen atom; (C3-C8)-cycloalkyl, (C2-C7)-heterocycloalkyl comprising 2 heteroatoms taking among nitrogen or oxygen atom; or (C6-C10)-aryl optionally substituted with one or more substitutes taken among (C1-C8)-alkoxy-group; X represents sulfur atom (S) or N(R4); Y represents nitrogen atom (N); R4 represents (C1-C8)-alkyl, phenyl-(C1-C8)-alkyl; or X represents sulfur atom (S), and Y represents CH; Z represents NH2 or OH; A represents sulfur (S), oxygen atom (O) or a bond. Also, invention relates to a pharmaceutical composition.

EFFECT: valuable properties of compounds and composition.

14 cl, 1 tbl, 119 ex

FIELD: medicine, gynecology.

SUBSTANCE: the present innovation deals with carrying out hormone-substitution therapy in women during menopause period or in sterilized women. For this purpose the above-mentioned therapy consists of the phase of relative dominant estrogen activity including three daytime dosages of a substance inducing estrogen activity being equivalent to approximately to 1 mg 17beta-estradiol daily, and the phase of relative dominant progestogen activity including combination of a substance inducing estrogen activity being equivalent to approximately 1 mg 17beta-estradiol daily and a substance that demonstrates progestogen activity being equivalent to approximately 90 mcg norgestimate daily. To fulfill such a therapy a pharmaceutical preparation is, also, suggested and a set of mentioned preparations. The innovation provides maximal weakening the symptoms of the disease, in particular, congestions along the safety of application due to decreased risk of known complications of estrogen therapy.

EFFECT: higher efficiency of therapy.

31 cl, 1 dwg, 1 ex, 2 tbl

FIELD: veterinary science.

SUBSTANCE: a sow should be twice injected with oxytocin and, additionally, intramuscularly about 2-4 h after afterbirth detachment one should introduce clathroprostin at the dosage of 1 ml. The innovation suggested is very efficient in preventing metritis-mastitis-agalactia and endometritis in sows, as well.

EFFECT: higher efficiency of prophylaxis.

1 ex, 1 tbl

The invention relates to medicine, namely, gynecology, and can be used to restore the fertility of women suffering from endometriosis

The invention relates to medicine, gynecology, and can be used for non-medical treatment of luteal phase in women

The invention relates to the derivatives of dihydronaphthalene formula I, where R1represents hydrogen, hydroxyl or alkyloxy, R2represents hydrogen, lower alkyl, aralkyl or phenyl, and R3represents pyridyl or imidazolyl

FIELD: medicine.

SUBSTANCE: method involves applying biphosphonates for producing preparations and treating osteoblastic (osteosclerotic) metastases connected with prostate cancer.

EFFECT: enhanced effectiveness in inhibiting abnormal osteoblast proliferation.

5 cl, 3 tbl

FIELD: medicine.

SUBSTANCE: method involves applying biphosphonates for producing preparations and treating osteoblastic (osteosclerotic) metastases connected with prostate cancer.

EFFECT: enhanced effectiveness in inhibiting abnormal osteoblast proliferation.

5 cl, 3 tbl

FIELD: medicine, oncology.

SUBSTANCE: through a catheter it is necessary to conduct prolonged polychemotherapy due to single regional intra-arterial injection of oxaliplatin at the dosage of 150 mg/sq. m into general hepatic artery as 12-h-long infusion and 5-fluorouracil at the dosage of 1000 mg/sq. m daily in the course of every course due to prolonged 120-h-long infusion. The duration of therapy - 4 courses of infusions at 28-d-long intervals. The innovation provides efficient therapy of the pathology mentioned due to increasing a single and a course dosage of anti-tumor preparations of low toxicity, at no complications that requires no hyperhydration procedures.

EFFECT: higher efficiency of therapy.

1 ex

FIELD: medicine, oncology.

SUBSTANCE: through a catheter it is necessary to conduct prolonged polychemotherapy due to single regional intra-arterial injection of oxaliplatin at the dosage of 150 mg/sq. m into general hepatic artery as 12-h-long infusion and 5-fluorouracil at the dosage of 1000 mg/sq. m daily in the course of every course due to prolonged 120-h-long infusion. The duration of therapy - 4 courses of infusions at 28-d-long intervals. The innovation provides efficient therapy of the pathology mentioned due to increasing a single and a course dosage of anti-tumor preparations of low toxicity, at no complications that requires no hyperhydration procedures.

EFFECT: higher efficiency of therapy.

1 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to derivatives of thiophene of the general formula (I): , wherein R1 is chosen from group consisting of hydrogen atom (H), -C(O)R7, -CO2R7, -C(O)NR7R8, -C(O)N(R7)OR8, -C(O)N(R7)-R2-OR8, -C(O)N(R7)-Ph, -C(O)N(R7)-R-Ph, -C(O)N(R7)S(O)2R8, -R2-OR7, -R2-O-C(O)R7, -C(S)R7, -C(S)NR7R8, -C(S)N(R7)-Ph, -C(S)N(R7)-R2-Ph, -R2-SR7, -CN, -OR7 and Het wherein Het represents tetrazolyl; Q1 represent group of the formula: -(R2)a-(Y1)b-(R2)c-R3 wherein a, b and a are similar or different and each means independently 0 or 1, and at least one among a or b means 1; n means 0, 1, 2, 3 or 4; Q2 represents group of the formula: -(R2)aa-(Y2)bb-(R2)cc-R4, or two adjacent Q2 groups represent -OR7 and in common with carbon atoms to which they are bound form 5-7-membered heterocycle comprising 1 or 2 heteroatoms chosen from oxygen atom (O); R5 is chosen from group consisting of H, alkyl and -NR7R8, or their pharmaceutically acceptable salts and solvates. Compounds can be used in treatment of states mediated by Polo-like kinase and sensitive neoplasm. Also, invention describes a method for synthesis of these compounds and preparing pharmaceutical compositions based on thereof.

EFFECT: valuable medicinal properties of compounds and pharmaceutical compositions.

27 cl, 1 tbl, 199 ex

FIELD: organic chemistry of natural compounds, medicine, pharmacy.

SUBSTANCE: invention relates to novel compounds of the formula (I): , wherein each R1, R2 and R3 means independently hydrogen atom or (C1-C4)-alkyl; R4 means (C1-C12)-alkyl optionally comprising from one to three substitutes chosen from group including hydroxy-group, (C1-C12)-alkoxycarbonyl, carbamoyl, (C2-C7)-alkenyl, (C6-C10)-aryl optionally comprising from one to three substitutes chosen from group including halogen atom, (C1-C12)-alkyl, (C1-C12)-alkoxy-, hydroxy-, (C1-C12)-alkylcarbonylamino-group, (C6-C10)-aryl-(C1-C12)-alkyl wherein aryl group comprises optionally from one to three substitutes chosen from group comprising halogen atom, (C1-C12)-alkyl, (C1-C12)-alkoxy-group, heterocyclyl-(C1-C12)-alkyl; R5 means hydroxy-, (C3-C7)-cycloalkylamino-group optionally substituted with phenyl, (C6-C10)-arylamino-, (C6-C10)-aryl-(C1-C4)-alkylamino-group optionally comprising from one to three substitutes chosen from group comprising sulfamoyl, (C1-C12)-alkyl, (C1-C12)-alkoxy-, hydroxy-group, heterocyclyl or benzyl, (C1-C4)-alkoxy-, benzhydrazino-group, heterocyclyl optionally comprising from one to three substitutes chosen from group including benzyl, benzhydryl, heterocyclylamino-group wherein heterocyclyl means saturated, unsaturated or aromatic monovalent cyclic radical comprising from 1 to 3 heteroatoms chosen from nitrogen (N), oxygen (O) and sulfur (S) atoms, or to their combination; n means a whole number 0, 1 or 2. Compounds of the formula (I) elicit anti-proliferative activity that allows their using in pharmaceutical composition. Also, invention describes intermediate compounds.

EFFECT: valuable medicinal properties of compounds and pharmaceutical composition.

30 cl, 1 tbl, 69 ex

FIELD: medicine.

SUBSTANCE: method involves carrying out 5-10 sessions of general magnetotherapy beginning from 3-rd day after surgical intervention in static mode: magnetic field rotation frequency is equal to 100 Hz, magnetic field intensity of 30 oersted, magnetic field shape is sinusoid half-cycle, raising time being equal to 30 s, time of recession 30 s; the number of procedures is equal to 15. Then, remote radiation therapy is administered 4 weeks later after surgical intervention. The treatment is applied to removed kidney bed and lymphatic collectors in single stage in classical fractioning mode at a doze of 2 Gy 5 times a week within 5 weeks up to reach total doze of 50 Gy with general magnetotherapy being concurrently applied in 1 session per day mode, the number of procedures being equal to 25. The first biotherapy course with recombinant human alpha tumor necrosis factor (TNF-α) is given at a dose of 2 mln units 2 weeks later after the radiation therapy being done. Total З biotherapy courses are to be given with 21 days long pause available between the courses.

EFFECT: enhanced effectiveness of treatment; reduced risk of postoperative complications; increased immune activity.

FIELD: medicine.

SUBSTANCE: method involves excising basic tumor as one-stage operation by means of high-energy СО2 laser or cytoreduction operation is carried out (basic tumor component removal) using laser output power of 20-40 W at the first stage. Remote photodynamic therapy is applied by irradiating the whole vulva surface with radiation dose of 100-150 J/cm2, power density of 50-70 mW/cm2 and irradiating tumor localization zone with radiation dose of 200-300 J/cm, power density of 150-200 mW/cm at the second stage. Additional interstitial laser irradiation with radiation dose of 200-300 J is optionally carried out at radiation power of 200-300 mW in tumor invasion zone.

EFFECT: enhanced effectiveness of organ-retaining and sparing treatment; accelerated treatment course; reduced risk of postoperative complications.

2 cl

FIELD: medicine, oncology.

SUBSTANCE: during the first day of therapy it is necessary to affect the lesion focus with alternating magnetic field at induction being 5-160 mTl, frequency of 50-100 Hz for about 7-10 min. Out of peripheral vein one should sample blood into two vials with hemoconservant per 100 ml/each; one vial should be supplemented with doxorubicin at the dosage of 50 mg/sq. m, the second vial - with 5-fluorouracil 750 mg/sq. m. Both vials should be incubated. Then it is necessary to inject intravenously by drops the content of the first, and then of the second vial for a patient. During the period since the first day up to the seventh one it is necessary to introduce cyclophosphan intramuscularly per 200 mg/sq. m. Since the first up to the tenth day of therapy one should affect with alternating and direct magnetic field. On the eighth day after seances of magnetic therapy one should repeatedly introduce anti-tumor chemopreparations at the same dosages and sequence. In two weeks it is necessary to repeat the above-mentioned curative impacts. In two weeks after the last introduction of chemopreparations one should fulfill surgical removal of lesion focus. The innovation enables to avoid side toxic manifestations of chemopreparations and notching of sutures and, also, healing due to secondary tension.

EFFECT: higher efficiency of therapy.

1 ex

FIELD: pharmaceutical industry, in particular agent having cytostatic and apoptosis-inducing activity.

SUBSTANCE: claimed agent represents β-Asparagine, obtained by extraction of ground burdock roots having specific particle size with heated water in specific raw materials/extractant ratio; decoction, extract separation, concentration up to dry residue in concentrate, filtering, and double recrystallization.

EFFECT: agent with high cytostatic and apoptosis-inducing (anti-tumor) activity.

5 dwg, 2 tbl

FIELD: medicine.

SUBSTANCE: the present innovation deals with the method of applying compositions containing trans-clomiphen and cis-clomiphen in the ratio by weight being above or equal 71/29 for increasing the level of testosterone in blood serum in male mammalian. Application of practically pure trans-clomiphen according to the present innovation provides increased level of testosterone both at the decrease and at sufficient content of this hormone, in case of necessity or desirability of such an increase for a mammalian, at decreased number of side effects as a result of clomiphen-based therapy.

EFFECT: higher efficiency of therapy.

4 cl, 3 dwg, 2 ex, 1 tbl

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