RU2302872C9 - Agent normalizing cartilage tissue function and method for its preparing - Google Patents

Abstract

FIELD: medicinal industry.

SUBSTANCE: invention relates to a method for isolation of biologically active substance from mammalian cartilage and to preparing a medicinal formulation for parenteral administration. Agent is made as a medicinal formulation for parenteral administration and represents peptide complex with the content of low-molecular fraction from 70% to 90%, molecular mass of its peptide components in the range 75-846 Da and concentration of polypeptides 2.5-2.9 mg/ml. Agent is prepared from cartilage of calves (age is 12 months, not above) or pigs by extraction with acetic acid in the presence of zinc chloride. Proposed method for preparing agent from cartilage of calves (age is 12 months, not above) or pigs involves tissue freezing at temperature -40°C (not less), keeping at temperature -20-22°C for 2 months (not less) followed by milling and addition of 3% acetic acid solution in the volume ratio = 1:5 at temperature 20 ± 5°C. Extraction is carried out at constant stirring up to preparation homogenous suspension and then 1% zinc chloride solution is added to suspension in the volume ratio = 50:1. Mixture is cooled at constant stirring up to temperature 7-16°C, stirred for 1 h in each 4 h settling for 48 h. Extract is separated from inert substances by separating and acetone is added to extract in the volume ratio = 1:5 and kept at temperature 3-5°C for 4 h. Formed homogenized deposit is precipitated with acetone repeatedly twice (not less) and deposit containing active substance is washed out on Nutch filter with two-fold volumes of acetone cooled to temperature 7-16°C up to preparing light-gray deposit. Deposit is rubbed through metallic sieve, dried, dissolved in distilled water at room temperature and constant stirring up to the concentration of polypeptides 2.5-2.9 mg/ml. Solution is centrifuged, filtered and subjected for ultrafiltration treatment in device at anti-pressure 1.0 kgf/cm² (not above) through materials with retaining capacity 15000 Da. Glycocol is added to ultrafiltrate up to its final concentration 10-20 mg/ml at pH = 5.6-6.6, solution is subjected for sterilizing filtration under pressure 2.0 kgf/cm² (not above), poured into ampoules in volume 2 ml and subjected for autoclaving at temperature 120°C for 8 min under atmosphere pressure 1.1 kgf/cm². Invention provides optimal technology in isolation of peptide complex with the content of low-molecular fraction from 70% to 90%, molecular mass of peptide components in the range from 75 to 846 Da and preparing aqueous extract with the concentration of polypeptides 2.5-2.9 mg/ml. Invention provides both purification of the end product from impurities and to enhance its yield. The isolated substance differs from the known substances prepared early from mammalian raw by molecular mass of peptide components, absence of toxicity and apyretic properties based on the complete removing impurities. Proposed substance can be used in medicine as an agent normalizing functions of cartilage tissue.

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

3 cl, 1 tbl, 1 dwg, 4 ex

Description

The invention 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 cartilage (sterno-costal joint) and the preparation of a parenteral dosage form that can be used in medicine as a means of normalizing the function of cartilage tissue.

Known methods for producing biologically active substances and medicines from animal raw materials (A.S. SU No. 1218521, 1994; A.S. SU No. 1227198, 1986; RF patent 1298879, 1993; RF patent No. 1448443, 1994; RF patent No. 1522486 , 1993; RF patent No. 2075944, 1997; RF patent No. 2161501, 2001; US patent No. 4341765; GB patent 1161896, 1969; FR patent 2583982, 1987).

It is known to use peptides in pharmaceutical compositions comprising these peptides to normalize cartilage tissue function (US patent No. 20072017554, 2002, A61K 038/17), which is the closest analogue to the proposed agent that normalizes cartilage tissue function.

Known methods for producing complex peptide preparations having tissue-specific action (RF patent No. 944191, 1994; RF patent No. 112226, 1993; RF patent No. 1417244, 1993; RF patent No. 2104702, 1998).

A known method of producing these drugs (Morozov V.G., Havinson V.Kh. Peptide bioregulators. St. Petersburg: Nauka, 1996. 74 pp.), Which includes obtaining a biologically active substance from animal organs and tissues by extraction with a 3% solution acetic acid with the addition of zinc chloride and the treatment of the supernatant with acetone, which is the closest analogue of the prototype for the proposed method of obtaining funds that normalize the function of cartilage tissue. The disadvantages of this method include the extraction during the extraction process from the tissue of a large amount (more than 70%) of substances of a non-peptide nature - ballast components, which, being impurities, do not determine the biological activity of the isolated active substance, as well as its low yield.

The present invention has posed and solved the problem of developing a method for producing a means that normalizes the function of cartilage, in the form of a parenteral dosage form isolated from cartilage (sterno-costal joint) of calves no older than 12 months of age or pigs, the technical result of which is the optimal technology of isolation a peptide complex with a low molecular weight fraction of from 70 to 90% with a molecular weight of its peptide components ranging from 75 to 846 Yes and obtaining aqueous extract solution with a concentration of the polypeptides of 2.5 ÷ 2.9 mg / ml, which allows not only to clean impurities from the resulting product, but also to increase its output.

Experimental verification showed that the proposed method provides pharmaceutical stability, maximum biological value and therapeutic efficacy of an agent that normalizes the function of cartilage, 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 the function of cartilage, in the form of a parenteral dosage form, which is a peptide complex with a low molecular weight fraction of 70 to 90%, with a molecular weight of its constituent peptide components ranging from 75 to 846 Yes, s the concentration of polypeptides 2.5 ÷ 2.9 mg / ml, the technical result of which is that the isolated substance differs from known substances obtained previously from animal raw materials in molecular weight (from 500 to 1500 Da) walking in his peptide components (RF №№1298879 patents 2,104,702, 2,163,129), as well as its non-toxicity and non-pyrogenic due to complete removal of impurities.

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

In the process of conducting research, the importance of the following stages of the method of obtaining funds normalizing the function of cartilage tissue, made in the form of a dosage form for parenteral administration (hereinafter referred to as the drug), was established:

- 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 normalizing the function of the cartilage tissue is made in the form of a parenteral dosage form and is a peptide complex with a low molecular weight fraction of 70 to 90%, with a molecular weight of the peptide components included in it within 75 to 846 Yes, with a concentration of polypeptides of 2.5 ÷ 2.9 mg / ml and obtained from cartilage (sterno-rib joint) of calves no older than 12 months of age or pigs by extraction with acetic acid in the presence of chloride about zinc.

The specified technical result is also achieved by the fact that the proposed method of obtaining a means of normalizing the function of cartilage tissue is characterized in that the cartilage (sterno-costal joint) of calves no older than 12 months of age or pigs are frozen at a temperature of at least minus 40 ° C, kept at temperature minus 20 ÷ 22 ° C for at least two months, then crushed, add 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 receiving about of a uniform 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 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 , acetone is added to the extract in a volume ratio of 1: 5, kept 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 a suction filter double volumes of acetone cooled to a temperature of 7 ÷ 16 ° С until a light gray precipitate is obtained, wiped through a metal sieve, dried, dissolved in distilled water at room temperature and with 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, pa the target is subjected to sterilizing filtration under a pressure of not more than 2.0 kgf / cm ² , poured into ampoules of 2 ml and autoclaved for 8 minutes at a temperature of 120 ° C and atmospheric pressure of 1.1 kgf / cm ² .

The invention is illustrated in the drawing and table.

The drawing shows the effect of the drug on the development of cartilage tissue explants.

The table shows the effect of the drug on the morphological and biochemical parameters of the peripheral blood of guinea pigs in the study of toxicity.

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

Cartilage (sterno-rib joint) 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.

300 l of a 3% solution of acetic acid are pumped into the extraction reactor and cooled to a temperature of (20 ± 5) ° С, then 60 kg of crushed material are 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 it is stirred for 1 hour every 4 hours of settling for 48 h

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 cartilage) is 30 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, mixed until the glycol is completely dissolved while maintaining a pH of 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 was poured into 2.0 mL ampules and autoclaved, wherein the drug vials are sterilized for 8 min at 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 (Serva, 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 75 to 846 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 75 to 846 Da, with a concentration of 2.5 polypeptides ÷ 2.9 mg / ml.

Example 2. The effect of the drug on the development of cartilage tissue explants

The experiments were carried out on 47 fragments of cartilaginous tissue of the proximal femur of the femur 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, glucose (0.6%), insulin (0.5 units / ml), penicillin (100 units / ml), glutamine (2 mm) were added to the medium. Cartilage fragments were placed in this medium and cultured on a collagen substrate in Petri dishes in a thermostat at a temperature of 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 cartilage tissue 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 cartilage tissue 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 31% was observed compared to the control PI values. When studying cartilage tissue 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 cartilage tissue, the drug exerted 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 72 white mongrel male mice with a body weight of 20–23 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 carried out on 60 white outbred male rats weighing 150 ÷ 180 g. Daily, once a day, animals of experimental groups were injected intramuscularly with doses of 1 mg / kg, 10 mg / kg, 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 intramuscular injection once daily for 6 months at 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 in a similar fashion 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), and erythrocyte resistance. In addition, the total protein content in blood serum was determined by the Lowry method, potassium and sodium by plasma spectrophotometry. After completion of the experiment, 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, 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 3 and 6 months after the start of the drug administration, no significant change in the indicators was revealed (table).

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 long-term administration to animals does not have toxic properties that prevent its further use as a medicine, made in the form of a dosage form for parenteral administration.

Example 4. The effectiveness of the drug in patients with osteochondrosis of the lumbar spine

The study was conducted with the participation of 33 patients with osteochondrosis of the lumbar spine at the age of 42 ÷ 59 years. Patients often noted the appearance of pain in the lower back with irradiation along the sciatic nerve, significantly intensifying with a change in body position, walking, physical activity. All patients previously received analgesics and anti-inflammatory drugs for a long time, the use of which caused a short-term therapeutic effect, requiring an increase in the dose of drugs for the course of treatment and their long-term use.

15 patients of the main group were injected with a dose of 5 mg in 2 ml of physiological saline solution intramuscularly once daily for 20 days. The control group included 18 patients who received injections of saline according to a similar pattern.

The effectiveness of the drug in patients was evaluated by the dynamics of clinical indicators and X-ray data.

It should be noted that the radiological symptoms of degenerative-dystrophic diseases of the spine are not only objective diagnostic criteria for the stage of development of the pathological process, but also have great prognostic significance during the ongoing drug therapy.

It was found that the use of the drug in patients with osteochondrosis of the lumbar spine during complex therapy contributed to a decrease in pain in 67.4% of cases. Such dynamics was most characteristic of middle-aged people. The progressive course of the disease with age, accompanied by characteristic radiological symptoms (narrowing of the lumen between adjacent vertebral bodies due to the flattening of degeneratively altered intervertebral discs, the formation of anterior and posterior osteophytes of the vertebral bodies, the presence of arthritic changes in the posterior and lateral intervertebral joints in the form of narrowing gaps, bumps, bumps the development of osteophytes along the edges of the articular ends, changes in the configuration of the intervertebral openings), contributes to the development of spondylosis and ondiloartroza and formation neurodystrophic and neurovascular syndromes. In these cases, the use of the drug in patients smoothed pain symptoms with a load on the spine and lower limbs. An improvement in the clinical picture of the course of the disease correlated with an improvement in objective criteria for X-ray diffraction patterns.

Thus, the results of the study confirm the normalizing effect of the drug in a therapeutically effective dose of 5 mg (2.5 mg / ml) on the function of cartilage and indicate the therapeutic efficacy of the drug and the feasibility of its use in the complex treatment of degenerative-dystrophic diseases of the spine at a dose of 5 mg intramuscularly once daily for 20 days.

A tool that normalizes the function of cartilage tissue, and a method for its preparation Indicator The introduction of the drug (10 mg / kg) 3 months 6 months Control (n = 24) Drug (n = 24) Control (n = 24) Drug (n = 24) Red blood cells, × 10 ¹² / l 5.2 ± 0.7 5.2 ± 0.4 5.3 ± 0.5 5.1 ± 0.4 Hemoglobin, g / l 14.2 ± 1.4 14.4 ± 1.3 14.2 ± 1.5 14.1 ± 1.7 Reticulocytes,% 1.3 ± 0.06 1.1 ± 0.05 1.2 ± 0.04 1.1 ± 0.07 Platelets × 10 ⁹ / L 142.7 ± 7.6 143.6 ± 6.4 142.8 ± 7.3 145.1 ± 7.9 White blood cells × 10 ⁹ / L 9.7 ± 0.6 9.6 ± 0.2 9.8 ± 0.2 9.6 ± 0.4 Band neutrons,% 0.32 ± 0.05 0.31 ± 0.03 0.30 ± 0.02 0.33 ± 0.04 Segmented neutrophils,% 43.2 + 2.3 44.7 ± 1.8 42.8 ± 2.2 44.7 ± 2.5 Eosinophils,% 0.69 ± 0.07 0.70 ± 0.05 0.67 ± 0.05 0.70 ± 0.04 Basophils,% 0.68 ± 0.04 0.69 ± 0.04 0.71 ± 0.05 0.72 ± 0.04 Monocytes,% 2.5 ± 0.06 2.4 ± 0.07 2.4 ± 0.08 2.5 ± 0.02 Lymphocytes,% 49.1 ± 2.5 50.3 ± 2.4 52.6 ± 2.1 52.3 ± 2.1 ESR, mm / h 1.85 ± 0.07 1.91 ± 0.03 1.91 ± 0.03 1.88 ± 0.06 Erythrocyte resistance,% NaCl - maximum 0.40 ± 0.03 0.41 ± 0.03 0.42 ± 0.04 0.42 ± 0.02 - minimum 0.32 ± 0.05 0.30 ± 0.05 0.31 ± 0.03 0.33 ± 0.05 Total protein in blood serum, g / l 72.7 ± 2.1 73.7 + 1.9 72.6 ± 2.5 71.9 + 2.7 Sodium in blood serum, mmol / l 153.6 ± 5.9 152.4 ± 6.2 154.8 ± 6.2 153.7 ± 5.9 Serum potassium, mmol / l 5.3 ± 1.7 5.2 ± 2.6 5.2 ± 2.5 5.1 ± 2.8

Claims (2)

1. A tool that normalizes the function of cartilage, characterized in that it is in the form of a parenteral dosage form and is a peptide complex with a low molecular weight fraction of from 70 to 90% with a molecular weight of its constituent peptide components ranging from 75 to 846 Yes, with a concentration of polypeptides of 2.5–2.9 mg / ml and obtained from cartilage (sterno-rib joint) of calves no 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 the function of cartilage, characterized in that the cartilage (sterno-rib joint) of calves no 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 50: 1 ratio, 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, kept 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 a suction filter with twice volumes of acetone cooled to a temperature of 7 ÷ 16 ° C to sieges and light gray in 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 backpressure 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 to gf / cm ² , poured into ampoules of 2 ml and autoclaved for 8 minutes at a temperature of 120 ° C and atmospheric pressure of 1.1 kgf / cm ² .