RU2303454C1 - Preparation for normalizing female reproductive function and method for its obtaining - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: the present innovation deals with isolating biologically active substance out of animals' ovaries to obtain medicinal form for parenteral intake that could be applied as a remedy to normalize female reproductive function. This preparation is being a peptide complex at the content of low-molecular fraction ranged 70-90%, at molecular weight of peptide components in it being within 70-712 Da, at concentration of polypeptides ranged 2.5÷2.9 mg/ml and obtained out of the ovaries of 12-mo-aged calves, not older or from swine due to extracting acetic acid in the presence of zinc chloride. The suggested technique for obtaining the preparation mentioned deals with the fact that ovaries from calves or swine's should be frozen at -40°C, not less to be kept at -20÷22°C for 2 mo, not less to be then reduced, 3%-acetic acid solution should be added at volumetric ratio of 1:5 at 20°±5°C; extraction should be fulfilled at constant mixing and on obtaining homogeneous suspension it should be supplemented with 1%-zinc chloride solution at volumetric ration of 7°÷16°C, then it is necessary to mix per 1 h in every 4 h keeping for 48 h, the extract should be separated against ballast substances due to separating, the extract should be supplemented with acetone at volumetric ratio of 1:5 to be kept at 3÷5°C for 4 h, the developed homogenized residue should be repeatedly precipitated with acetone twice, not less, then the residue that contains an active substance should be washed upon Nutsch filter with 2-fold volumes of acetone (cooled up to 7÷16°C) till obtaining the residue of light-gray color, grated through metallic sieve, dried, dissolved in distilled water at a room temperature and constant mixing up to polypeptides concentration being 2.5÷2.9 mg/ml,; the solution should be centrifuged, filtered and purified ultrafiltrationally upon the equipment at anti-pressure being not more than 1.0 kilogauss/sq. cm through materials at inhibiting capacity of 15000 Da, ultrafiltrate should be supplemented with glycocoll up to its final concentration being 10÷20 mg/ml at pH=5.6÷6.6, the solution should undergo sterilizing filtration at the pressure being not more than 2.0 kilogauss/sq. cm to be poured into ampoules per 2 ml and autoclaved for 8 min at 120°C and atmospheric pressure of 1.1 kilogauss/sq. cm. The innovation provides optimal technology to isolate peptide complex at the content of low-molecular fraction from 70 to 90% at molecular weight of peptide components in it ranged 70-712 Da and obtain water solution of the extract at concentration of polypeptides being 2.5÷2.9 mg/ml that enables not only purify the obtained product against admixtures but increase its output; and, also, the isolated substance is distinct against known ones obtained earlier out of animal ovaries by molecular weight of peptide complexes in it and by its nontoxicity and apyrogeneity due to complete purification against admixtures.

EFFECT: higher efficiency.

2 cl, 1 dwg, 5 ex, 2 tbl

Description

The group of inventions relates to a medicinal product and a method for its preparation from animal organs, in particular to the isolation of a biologically active substance from the ovaries and the preparation of a parenteral dosage form that can be used in medicine as a means of normalizing reproductive function in women.

Known methods for producing biologically active substances from organic raw materials (urine of pregnant women, urine of postmenopausal women, blood serum of mares) in order to restore reproductive function in women (Gemzell S.A., Diczjalusy E., Tillinger KG Clinical effect of human pituitary follicle stimulating hormone (FSH). - J, Clin. Metab. - 1958. Vol. 18. - pp. 1333 ÷ 1342).

The disadvantages of known substances include the fact that these substances have a pronounced hormonal effect, give a short-term effect and numerous complications (endocrine disorders, multiple pregnancy, etc.).

Known substance that restores the reproductive function obtained from the ovaries of animals (RF patent No. 2056853, 1993, AK 35/48), which is the closest analogue - the prototype for the proposed tool and method for its preparation.

According to the method described in the patent of the Russian Federation No. 2056853, the processing of crushed ovaries of animals is carried out with acetic acid, at pH 2.5 ÷ 4.0, for 24 ÷ 36 hours, at a temperature of 0 ÷ 4 ° C with the addition of 0.3 ÷ 0, 7 g / l zinc chloride. Polypeptides are precipitated with acetone in an amount of 8-12 volumes per 1 volume of extract at a temperature of minus 2 ÷ minus 4 ° C for 10-16 hours. The precipitate is separated by filtration, dried, and then redissolved in distilled water, filtered and lyophilized. This method allows to obtain a complex of polypeptides with a molecular weight of 1000-10000 Yes.

The disadvantages of this method include the extraction during the extraction process from the ovarian tissue of a large amount (more than 70%) of non-peptide substances, which, being impurities, do not determine the biological activity of the isolated active substance, as well as its low yield.

The present invention posed and solved the problem of developing a method for producing a product that normalizes reproductive function in women in the form of a parenteral dosage form isolated from ovaries of calves no older than 12 months of age or pigs, the technical result of which is the optimal technology for isolating a peptide complex with a low molecular weight content fractions from 70 to 90% with a molecular weight of its peptide components ranging from 70 to 712 Yes and obtaining an aqueous solution of the extract with a concentration of polypeptides of 2.5 ÷ 2.9 mg / ml, which allows not only to clean the resulting product from impurities, but also to increase its yield.

Experimental verification showed that the proposed method provides pharmaceutical stability, maximum biological value and therapeutic efficacy of a product that normalizes reproductive function in women, made in the form of a parenteral dosage form, which is confirmed by the experimental results given in the examples illustrating the invention.

Another aspect of the invention is a means of normalizing reproductive function in women, in the form of a parenteral dosage form, which is a peptide complex with a low molecular weight fraction from 70 to 90% with a molecular weight of its constituent peptide components ranging from 70 to 712 Yes, s the concentration of polypeptides 2.5 ÷ 2.9 mg / ml, the technical result of which is that the isolated substance differs from the known substances obtained previously from the ovaries of animals in molecular weight (from 1 000 to 10000 Yes) of the peptide components included therein (RF patent No. 2056853), as well as its non-toxicity and pyrogen-free nature due to complete purification from impurities.

In the course of studies in animal experiments, it was found that the obtained aqueous solution of the drug with a concentration of polypeptides of 2.5 ÷ 2.9 mg / ml is a therapeutically effective dose, which allows us to consider the use of the drug for various types of pathology of the reproductive system in women. This is confirmed by the following examples.

In the process of conducting research, the importance of the following stages of the method for producing a product that normalizes reproductive function in women was established in the form of a dosage form for parenteral administration - (hereinafter - the drug):

- repeated precipitation of the resulting homogenized precipitate with double volumes of acetone at least two times and subsequent washing on the suction filter with double volumes of acetone until a light gray precipitate is obtained, which indicates complete purification of the precipitate from impurities such as lipid, protein, phospholipid and others;

- obtaining an aqueous solution of the extract in a concentration of polypeptides of 2.5 ÷ 2.9 mg / ml; its centrifugation, filtration, followed by ultrafiltration cleaning at the installation with a back pressure of not more than 1.0 kgf / cm ² through materials with a retention capacity of 15,000 Da and the addition of glycocol to the ultrafiltrate to a final concentration of 10 ÷ 20 mg / ml at pH + 5.6 ÷ 6.6;

- sterilizing filtration, ampouling and autoclaving for 8 minutes at a temperature of 120 ° C and atmospheric pressure of 1.1 kgf / cm ² , which ensures the stability of the drug while maintaining therapeutic efficacy.

The mode and time of autoclaving were established experimentally.

The specified technical result is achieved in that the agent that normalizes the reproductive function in women is made in the form of a parenteral dosage form and is a peptide complex with a low molecular weight fraction from 70 to 90% with a molecular weight of the peptide components included in it within 70 to 712 Yes, with a concentration of polypeptides of 2.5 ÷ 2.9 mg / ml and obtained from the ovaries of calves not older than 12 months of age or pigs by extraction with acetic acid in the presence of zinc chloride.

The specified technical result is also achieved by the fact that the proposed method of obtaining a means of normalizing reproductive function in women is characterized in that the ovaries of calves not older than 12 months of age or pigs are frozen at a temperature of at least minus 40 ° C, kept at a temperature of minus 20 ÷ 22 ° C for at least two months, then crushed, add a 3% solution of acetic acid in a volume ratio of 1: 5 at a temperature of 20 ° ± 5 ° C, extraction is carried out with constant stirring, after obtaining a homogeneous suspension in it is added a 1% solution of zinc chloride in a volume ratio of 50: 1, cooled with constant stirring to a temperature of 7 ° ÷ 16 ° C, then stirred for 1 hour after every 4 hours of sedimentation for 48 hours, the extract is separated from the ballast substances by separation, to acetone is added to the extract in a volume ratio of 1: 5, kept at a temperature of 3 ÷ 5 ° C for 4 hours, the resulting homogenized precipitate is re-precipitated with acetone at least 2 times, then the precipitate containing the active substance is washed with a suction filter in double volumes cooling of acetone to a temperature of 7 ° to 16 ° C to obtain a light gray precipitate, wiped through a metal sieve, dried, dissolved in distilled water at room temperature and constant stirring to a concentration of polypeptides of 2.5 ÷ 2.9 mg / ml, solution centrifuged, filtered, subjected to ultrafiltration cleaning at the installation with a backpressure of not more than 1.0 kgf / cm ² through materials with a retention capacity of 15,000 Yes, glycol is added to the ultrafiltrate to a final concentration of 10 ÷ 20 mg / ml at pH = 5.6 ÷ 6 , 6, the solution subjected They sterilize the filtration under a pressure of not more than 2.0 kgf / cm ² , pour it into 2 ml ampoules and autoclave it for 8 minutes at a temperature of 120 ° C and atmospheric pressure of 1.1 kgf / cm ² .

The invention is illustrated by the figure and tables.

The drawing shows the effect of the drug on the development of ovarian explants.

Table 1 presents the effect of the drug on the morphological and biochemical parameters of the peripheral blood of guinea pigs in the study of toxicity.

Table 2 shows the effect of the drug on the level of serum adenohypophysis hormones in patients with menopausal syndrome.

The invention is illustrated by the following examples: example 1 - a method of obtaining a preparation; example 2, confirming the biological activity of the drug; example 3 - the study of the toxicity of the drug; example 4, confirming the therapeutic effectiveness of the drug, made in the form of a dosage form for parenteral administration.

Example 1. The method of obtaining the drug

The ovaries of calves (not older than 12 months of age) or pigs, which are frozen at a temperature of at least minus 40 ° C and kept at a temperature of minus 20 ÷ 22 ° C for at least two months, are used as raw materials.

350 l of a 3% solution of acetic acid are pumped into the extraction reactor and cooled to a temperature of (20 ± 5) ° С, then 70 kg of crushed material is loaded into the reactor with constant stirring until a homogeneous mass of raw material is obtained. The extraction is carried out with constant stirring, after obtaining a homogeneous suspension, a 1% solution of zinc chloride in a volume ratio of 50: 1 is added to it, cooled with constant stirring to a temperature of plus 7 ÷ 16 ° C, then stirred for 1 hour every. 4 hours of settling for 48 hours. The extract is separated from the ballast substances by separation at (5000 ± 500) rpm for 1 hour. Acetone in a volume ratio of 1: 5 is added to the extract. It is kept at a temperature of plus 3 ÷ 5 ° C for 4 hours. The precipitate formed is homogenized and re-precipitated with acetone at least two times. Then the precipitate containing the active substance is washed on a suction filter with twice volumes of acetone cooled to a temperature of 7-16 ° C until a light gray precipitate is obtained. The washed precipitate is wiped through a metal sieve, spread in a thin layer into enameled cuvettes, covered with a double layer of cotton cloth and dried with periodic stirring in a fume hood until the smell of acetone is completely removed.

The yield of active substance (powder of a biologically active peptide complex isolated from the ovaries) is 60 g per 1 kg of feedstock.

The resulting powder is dissolved in distilled water with constant stirring at room temperature for 40 min to a concentration of peptides 2.5 ÷ 2.9 mg / ml The resulting solution was centrifuged at (3000 ± 200) rpm for (20 ± 5) minutes. The centrifuge is filtered through an AP-15 filter or similar.

The filtrate is subjected to ultrafiltration purification in an ultrafiltration unit with a backpressure of not more than 1.0 kgf / cm ² , through materials with a retention capacity of 15,000 Da.

A calculated sample of glycocol is added to the ultrafiltrate, to its final concentration of 10–20 mg / ml, and mixed until the glycol is completely dissolved while maintaining pH = 5.6–6.6.

The solution is subjected to sterilizing filtration, which is carried out under pressure not exceeding 2.0 kgf / cm ² .

The resulting solution is poured into 2.0 ml ampoules and autoclaved, and the ampoules with the preparation are sterilized for 8 minutes at a temperature of 120 ° C and atmospheric pressure of not more than 1.1 kgf / cm ² .

The preparation is a colorless, transparent solution and contains a peptide complex with a concentration of polypeptides of 2.5 ÷ 2.9 mg / ml.

Testing for the absence of high molecular weight protein components is carried out by adding to the contents of 1 ampoule of the drug 1 ml of a 10% solution of trichloroacetic acid. The transparency of the solution indicates the absence of high molecular weight protein components.

To determine peptide bonds in a preparation, a biuret reagent is added to its solution. Staining the solution with violet color indicates the peptide bonds present in the preparation.

In order to determine the polypeptides and their fractions in the preparation, the methods of ultraviolet spectrophotometry, gel chromatography, mass spectrometry, high performance liquid chromatography and polyacrylamide gel electrophoresis are used.

The ultraviolet spectrum of the drug solution is recorded in the region of wavelengths from 250 to 350 nm: the absorption maximum is observed at a wavelength of 270 ± 5 nm.

The molecular weight of the polypeptides that make up the drug is determined by the following methods:

by gel chromatography on Sephadex G-25 and G-50 (Pharmacia, Sweden). Peptide Molecuar Weight Kit MS III (Sen / a, Germany) was used to calibrate a 1.6 x 60 cm column;

mass spectrometry method. Spectra are obtained on a Voyager DE Biospectrometry time-of-flight mass spectrometer with laser desorption and ionization using a matrix (MALDI-TOF) at a relative intensity of a nitrogen laser of 2300 ÷ 2400, an accelerating voltage of 25000 V, a delay time of 90 ns, and a pressure in the vacuum chamber of 2.6 × 10 ⁶ torr;

by electrophoresis in a 15% polyacrylamide gel in comparison with a standard set of marker proteins.

Using the methods listed above, it was found that the composition of the drug includes polypeptides with a molecular weight of from 70 to 712 Da.

Using reverse-phase high-performance liquid chromatography in an acetonitrile gradient (Lichrosorb C18 sorbent, column 2 × 62 mm), it was found that the preparation contains mainly low molecular weight fractions - from 70 to 90%, and high molecular weight components are absent in the preparation.

Pyrogenicity of the drug is determined in rabbits by the generally accepted method (GF XI, issue 2, p. 183) with a test dose of 0.25 mg per 1 kg of animal weight in 1.0 ml of isotonic 0.9% sodium chloride solution for injection. It is shown that the drug is pyrogen-free.

Thus, in the above method, a preparation is obtained - a parenteral dosage form, which is a peptide complex with a low molecular weight fraction from 70 to 90%, with a molecular weight of its peptide components ranging from 70 to 712 Da, with a concentration of 2.5 polypeptides ÷ 2.9 mg / ml.

Example 2. The effect of the drug on the development of ovarian explants

The experiments were carried out on 45 fragments of ovaries of Wistar rats weighing 150–200 g. The culture medium for explant cultivation consisted of 35% Eagle's solution, 25% fetal calf serum, 35% Hanks solution, 5% chicken embryonic extract, on Wednesday glucose (0.6%), insulin (0.5 units / ml), penicillin (100 units / ml), glutamine (2 mM) were added. Ovarian fragments were placed in this medium and cultured on a collagen substrate in Petri dishes in an incubator at 36.7 ° C for 2 days. The preparation was added to the experimental medium at concentrations of 1, 10, 50, 100, 200, and 400 ng / ml. The criterion of biological activity was the area index (PI) - the ratio of the area of the entire explant along with the growth zone to the outgoing area of the ovarian fragment. The IP values were expressed as a percentage, the control IP value was taken as 100%.

The drawing shows the effect of the drug on the development of ovarian explants. It was found that after 1 day of cultivation, the explants spread on a collagen substrate and the proliferation of proliferating and migrating cells began on the periphery of the explant. On the 3rd day of cultivation at a drug concentration of 50 ng / ml, a significant increase in the explant PI by 41% was observed, compared with the reference PI values. In the study of ovarian explants for longer periods of cultivation (7 days), a similar stimulating effect of the drug at the same concentration was revealed.

Thus, in relation to the ovaries, the drug had a tissue-specific effect, manifested in stimulating the growth of explants.

Example 3. The study of the toxicity of the drug

The general toxic effect of the drug was investigated in accordance with the requirements of the Guidelines for the experimental (preclinical) study of new pharmacological substances (2000): acute toxicity with a single administration of the drug, as well as subacute and chronic toxicity with prolonged administration of the drug.

A study of acute toxicity was conducted on 66 white outbred male mice weighing 18-20 g. Animals were randomly divided into 6 equal groups. The drug was administered to animals once intramuscularly at doses of 35 mg / kg, 50 mg / kg, 100 mg / kg, 150 mg / kg, 200 mg / kg in 0.25 ml of sterile 0.9% NaCl solution. Animals of the control group were injected with the same volume of 0.9% NaCl solution.

Studies on the study of subacute toxicity were performed on 70 white outbred male rats weighing 170–190 g. The animals were administered intramuscularly daily to animals of experimental groups for 90 days at doses of 1 mg / kg, 10 mg / kg, and 100 mg / kg 0.5 ml of a sterile 0.9% NaCl solution. The animals of the control group were injected in the same volume with a sterile 0.9% NaCl solution. Before administration of the drug, on the 30, 60 and 90 days after the start of drug administration, the morphological composition and properties of peripheral blood were studied in animals. At the end of the experiment, biochemical and coagulological blood parameters were examined.

Studies on the study of chronic toxicity were carried out for 6 months, based on the duration of the recommended clinical prescription of the drug, in 84 male guinea pigs weighing 250-280 g. Animals of the experimental groups received once daily intramuscular preparation for 6 months. in doses of 1 mg / kg, 10 mg / kg, 100 mg / kg in 0.5 ml of a sterile 0.9% NaCl solution. In the control group, animals were injected using a similar scheme with a sterile 0.9% NaCl solution in the same volume. In animals, peripheral blood was determined by conventional methods:

the number of red blood cells, hemoglobin, reticulocytes, platelets, white blood cells, white blood cell count, erythrocyte sedimentation rate (ESR), erythrocyte resistance. Along with this, the content of total protein in blood serum was determined by the method of Lowry, potassium and sodium by plasma spectrophotometry. After the experiment was completed, a pathomorphological study of the brain and spinal cord, spinal ganglia, thyroid gland, parathyroid glands, adrenal glands, testes, pituitary gland, heart, lungs, aorta, liver, kidney, bladder, pancreas, stomach, small intestine, colon intestines, thymus, spleen, lymph nodes, bone marrow.

When studying acute toxicity, it was found that a single administration of the studied drug to animals in a dose exceeding the therapeutic recommended for clinical use by more than 5000 times does not cause toxic reactions, which indicates a large therapeutic breadth of the drug.

The study of subacute and chronic toxicity of the drug indicates the absence of side effects with prolonged use of the drug in doses exceeding the therapeutic 300-3000 times. When studying the effect of the drug on the morphological composition and biochemical parameters of the peripheral blood of guinea pigs after 3 and 6 months after the start of drug administration, no significant change in the indicators was revealed (Table 1).

When assessing the general condition of animals, morphological and biochemical parameters of peripheral blood, the morphological state of internal organs, the state of the cardiovascular and respiratory systems, and liver and kidney function, pathological changes in the body were not detected.

Therefore, the drug obtained by the proposed method, with prolonged administration to animals does not have toxic properties that impede its further use as a medicinal product made in the form of a parenteral dosage form.

Example 4. The effectiveness of the drug in women with menopause

A study of the effectiveness of the drug was carried out with the participation of 43 women aged 45 ÷ 55 years with menopausal syndrome. Patients complained of hot flashes to the head and upper body, hyperhidrosis, increased irritability, emotional lability. Among objective symptoms, signs of colpitis, urethrocystitis, vulvar atrophy predominated. In a number of cases, symptoms of osteoporosis were noted.

Patients were randomly divided into 2 groups. The main group included 28 patients who were injected with the drug at a dose of 5 mg in 2.0 ml of saline intramuscularly once daily for 10 days.

The control group included 15 women who received injections of saline according to a similar pattern. Patients who had previously received hormone replacement therapy were not included in the study.

Complaints of patients were evaluated in dynamics, a general clinical study of blood and urine, a biochemical study of blood, an ultrasound examination of the ovaries were performed. The content of hormones (ACTH, TSH, FSH and LH) in the blood serum was determined by the radioimmunological method.

It was established that the use of the drug in patients with menopausal syndrome contributed to an improvement in the general condition, which was manifested in a decrease in the number of hot flashes, improved sleep, appetite, and increased performance.

In a laboratory study of hormonal status indicators, a significant decrease in the FSH content was noted, which caused an increase in the LH / FSH index to the lower boundaries of age-related physiological fluctuations (Table 2). There was also a downward trend in ACTH and TSH.

Normalization of hormonal parameters in patients of the main group after the use of the drug was correlated with an improvement in the clinical manifestations of menopausal syndrome and indicated the restoration of an adequate adaptive reaction of an aging organism in response to an age-related decrease in ovarian function. The study revealed no side effects and complications.

Thus, the results of a clinical trial indicate the therapeutic efficacy of the drug and the feasibility of its use in the complex treatment of menopausal syndrome in an effective therapeutic dose of 5 mg (2.5 mg / ml) intramuscularly once daily for 10 days.

Table 1 A tool that normalizes reproductive function in women, and a method for its preparation Indicator The introduction of the drug (10 mg / kg) 3 months 6 months Control (n = 21) Drug (n = 21) Control (n = 21) Drug (n = 21) Red blood cells, × 10 ¹² / l 5.4 ± 0.2 5.2 ± 0.4 5.3 ± 0.2 5.1 ± 0.4 Hemoglobin, g / l 14.3 ± 1.7 14.1 ± 1.5 14.2 ± 1.8 14.2 ± 1.9 Reticulocytes,% 1.1 ± 0.05 1.2 ± 0.03 1.3 ± 0.06 1.1 ± 0.08 Platelets × 10 ⁹ / L 142.1 ± 5.2 141.3 ± 5.3 140.4 ± 4.1 143.8 ± 4.6 White blood cells × 10 ⁹ / L 9.6 ± 0.1 9.5 ± 0.2 9.4 ± 0.2 9.6 ± 0.4 Band neutrons,% 0.35 ± 0.05 0.32 ± 0.05 0.33 ± 0.04 0.34 ± 0.07 Segmented neutrophils,% 42.7 ± 2.7 41.6 ± 2.8 43.6 ± 2.5 44.1 ± 2.3 Eosinophils,% 0.74 ± 0.02 0.76 ± 0.03 0.74 ± 0.04 0.75 ± 0.05 Basophils,% 0.62 ± 0.03 0.62 ± 0.06 0.65 ± 0.03 0.62 ± 0.07 Monocytes,% 2.5 ± 0.01 2.4 ± 0.02 2.7 ± 0.03 2.4 ± 0.03 Lymphocytes,% 44.7 ± 2.3 48.3 ± 2.5 43.8 ± 2.7 45.7 ± 1.9 ESR, mm / h 1.83 ± 0.06 1.76 ± 0.07 1.86 ± 0.04 1.78 ± 0.04 Erythrocyte resistance,% NaCl - maximum 0.45 ± 0.03 0.44 ± 0.02 0.43 ± 0.07 0.42 ± 0.03 - minimum 0.35 ± 0.02 0.33 ± 0.02 0.34 ± 0.05 0.35 ± 0.05 Total protein in blood serum, g / l 75.2 ± 1.8 76.2 ± 1.7 75.8 ± 2.1 74.3 ± 2.3 Sodium in blood serum, mmol / l 150.1 ± 5.7 151.3 ± 5.8 152.5 ± 6.2 149.4 ± 5.4 Serum potassium, mmol / l 5.0 ± 1.9 5.2 ± 2.4 5.3 ± 1.9 5.3 ± 2.2

table 2 A tool that normalizes reproductive function in women, and a method for its preparation Indicator Before treatment After treatment in the control group After treatment with the drug FSH, ng / ml 72.5 ± 3.6 68.4 ± 3.3 34.5 ± 3.7 ^* LH, ng / ml 10.1 ± 0.8 10.2 ± 1.1 10.9 ± 0.9 ^* - P <0.05 compared with the indicator before treatment.

Claims (2)

1. A tool that normalizes reproductive function in women, characterized in that it is made in the form of a parenteral dosage form and is a peptide complex with a low molecular weight fraction from 70 to 90% with a molecular weight of the peptide components included in it ranging from 70 to 712 Yes, with a concentration of polypeptides of 2.5 ÷ 2.9 mg / ml and obtained from the ovaries of calves not older than 12 months of age or pigs by extraction with acetic acid in the presence of zinc chloride. 2. A method of obtaining a means of normalizing reproductive function in women, characterized in that the ovaries of calves not older than 12 months of age or pigs are frozen at a temperature of at least minus 40 ° C, kept at a temperature of minus 20 ÷ 22 ° C for at least two months, then crushed, add a 3% solution of acetic acid in a volume ratio of 1: 5 at a temperature of 20 ° ± 5 ° C, the extraction is carried out with constant stirring, after obtaining a homogeneous suspension, a 1% solution of zinc chloride in volume is added to it correlation 50: 1, cooled with constant stirring to a temperature of 7 ÷ 16 ° C, then stirred for 1 hour after every 4 hours of settling for 48 hours, the extract is separated from the ballast substances by separation, acetone is added to the extract in a volume ratio of 1: 5, maintained at a temperature of 3 ÷ 5 ° C for 4 h, the resulting homogenized precipitate is re-precipitated with acetone at least 2 times, then the precipitate containing the active substance is washed on the suction filter with twice volumes of acetone cooled to a temperature of 7 ÷ 16 ° C until a precipitate is obtained light gray color, wiped through a metal sieve, dried, dissolved in distilled water at room temperature and constant stirring to a concentration of polypeptides of 2.5 ÷ 2.9 mg / ml, the solution is centrifuged, filtered, subjected to ultrafiltration purification on the installation with a backpressure of not more than 1, 0 kgf / cm ² through materials with a retention capacity of 15,000 Yes, glycol is added to the ultrafiltrate to its final concentration of 10 ÷ 20 mg / ml at pH 5.6 ÷ 6.6, the solution is subjected to sterilizing filtration under a pressure of not more than 2.0 kgf / ^cm2 dec vayut into vials of 2 ml and autoclaved for 8 minutes at 120 ° C and atmospheric pressure of 1.1 kgf / cm ^2.