RU2290936C1 - Method for preparing agent for maintaining therapy possessing tissue-specific activity - Google Patents

Abstract

FIELD: medicine, peptides.

SUBSTANCE: invention relates to a method for preparing a peptide complex from animal raw possessing the tissue-specific activity. Method for preparing an agent for maintaining therapy possessing tissue-specific activity involves milling calf or pig organs of age 12 months, not above, addition of 3% acetic acid solution at temperature 20 ± 5°C and extraction is carried out at constant stirring. Then, in 30 min zinc chloride 1% solution is added, mixture is cooled to temperature 7-16°C at constant stirring followed by stirring for every 1 h in each 4 h and settling for 48 h. Extract is separated from inert substances by separation and acetone is added to extract in the volume ratio = 1:5, mixture is kept at 3-5°C for 4 h and formed precipitate is washed out with out with two-fold volume of acetone cooled to temperature 7-16°C, formed precipitate is rubbed through a metallic sieve and prepared end product is dried at temperature 18 ± 2°C. The end product represents a peptide complex with the content of low-molecular peptide fraction from 70% to 90% and with molecular mass of its peptide components in the range 1000-12000 Da wherein this complex comprises amino acids, mineral substances, trace elements and vitamins in biologically bound form. The complex elicits the expressed tissue-specific activity based on the proposed sequence of technological procedures and conditions for their realization involving temperature, temporal and other indices, and by using substances including the parent raw, a definite extractant and others. Peptide component in the prepared complex has no denaturating properties and retains its regulatory properties that suggests its using as an agent for carrying out the maintaining therapy. The proposed agent can be used in medicinal practice as an agent used in carrying out the maintaining therapy.

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

9 cl, 42 tbl, 14 ex

Description

The invention relates to medicine, and relates to the production of peptide complex from animal raw materials with tissue-specific activity, which may find application in medical practice as a means for maintenance therapy.

A known method of producing peptides from animal raw materials, namely from the pineal gland of cattle (RF patent No. 2136296, 1997), which is crushed, extracted in a solution of acetic acid, frozen at a temperature of minus 5-20 ° C and thawed, after which the pre-clarified extract subjected to ultrafiltration in a tangential flow on membranes with a transmission threshold of 10 kD. The yield of the target product from a unit of raw material is 12 g per 1 kg of feedstock. The resulting peptide complex according to the method described in RF patent No. 2136296 does not contain protein contamination (negative reaction to a protein with trichloroacetic acid, as well as data from high performance liquid chromatography), meets the criteria of authenticity (maximum absorption in the UV spectrum is 270 ± 5 nm) and has specific activity.

A known method of producing a peptide complex from the pineal gland of cattle (RF patent No. 944191, 1994), including extraction of crushed raw materials with 3-5% acetic acid with the addition of zinc chloride, centrifugation, sedimentation of the target product from the supernatant with six volumes of acetone and ether, its renaturation in 1% acetic acid, followed by clarification and preparation of the dosage form.

However, the peptide complex obtained in this way contains protein contamination, and the yield of the target product from a unit of raw material is small and amounts to 7 g / kg.

The closest technical solution is a method of obtaining from animal raw materials having a tissue-specific action of a nucleoprotein complex (RF patent No. 2075944, 1996) with a molecular mass of its constituent nucleoprotein components from 10,000 to 90,000 Da.

The method described in the patent of the Russian Federation No. 2075944, provides for the extraction of pre-prepared (crushed and frozen) animal raw materials using as extractant a 1-2.5% solution of sodium carbonate (soda) at pH 10.4-10.8 for 2- 4 hours with the addition of 0.1-0.5% magnesium chloride with a ratio of the volumes of extracted tissue and soda solution in the range of 1: 8-1: 20, separation of the precipitate by filtration or in a centrifuge, acidification of the extract to pH 3.8-5 5, cooling the suspension at a temperature not exceeding 4 ° C for 40 hours to remove insoluble substances, washing the precipitate with organic solvents (sequentially ethanol, acetone, sulfuric ether), drying the precipitate in vacuum at a temperature not exceeding 55 ° C.

The yield of the target product (powder of the nucleoprotein complex) is 2.5-3.0% of the amount of feedstock.

It should be noted that although the target product obtained by the method described in RF patent No. 2075944 contains nucleoprotein complexes with specificity of action and tropism with respect to organs and tissues, which are the source of their production, the known method has a number of disadvantages, among which it should be noted the following.

The powder of the nucleoprotein complex from organs and tissues obtained by the described method belongs to the group of sparingly soluble substances, which requires certain technological methods for use in medical practice.

The resulting product consists of two groups of molecular associates: a sufficiently large number of high molecular weight components, among which there are proteins of the amyloid class, which include prions (mol. Wt. From 27,000 to 31,000 Da), as well as a nucleic component, the presence of which is dangerous the presence of proto-oncogenes in the target product.

The problem to which the claimed invention is directed is to develop a technology for the isolation of a low molecular weight complex purified from protein, nucleic and lipid impurities, the peptide component of which does not denature and retains its regulatory properties.

The technical result consists in the fact that the proposed method produces a peptide complex with a molecular weight of its peptide components from 1000 to 12000 Yes, containing amino acids, minerals, trace elements and vitamins in a biologically bound form, which exhibits pronounced tissue-specific activity, as a result of which the peptide component of the obtained complex does not denature and retains its regulatory properties, which allows us to consider its use as a means for supporting t rapii.

The method of obtaining funds for maintenance therapy with tissue-specific activity, includes grinding raw materials of animal origin, extraction, which is carried out with a 3% solution of acetic acid with the addition of zinc chloride, cooling to a temperature of plus 7-16 ° C with constant stirring for 48 hours, separation of the extract from ballast substances by separation, precipitation, which is carried out with acetone in a volume ratio of 1: 5 at a temperature of plus 3-5 ° C for 4 hours, sedimentation until a formed precipitate is formed, which is then washed with twice volumes of acetone, the washed precipitate is wiped through a metal sieve, the obtained target product is dried at a temperature of 18 ± 2 ° C until the traces of acetone are completely removed.

Moreover, as a target product, a peptide complex with a molecular weight of the peptide components included in it is obtained in the range from 1000 to 12000 Da.

In this case, the liver is used as animal raw material, and the peptide complex that supports the liver function contains amino acids, minerals, trace elements and vitamins in a biologically bound form in the following ratio of components, wt.%:

amino acids 65.0-75.0 minerals 3.0-5.0 trace elements 0.003-0.008 vitamins 0.01-0.03

In this case, the pancreas is used as animal raw material, and the peptide complex supporting the pancreas function contains amino acids, minerals, trace elements and vitamins in a biologically related form in the following ratio of components, wt.%:

amino acids 45.0-55.0 minerals 10.0-15.0 trace elements 0.001-0.003 vitamins 0.005-0.010

At the same time, the thyroid gland is used as animal raw material, and the peptide complex that supports the thyroid gland function contains amino acids, minerals, trace elements and vitamins in a biologically bound form in the following ratio of components, wt.%:

amino acids 70.0-80.0 minerals 3.0-5.0 trace elements 0.002-0.004 vitamins 0.001-0.003

In this case, vessels are used as animal raw materials, and the peptide complex that supports the function of vessels contains amino acids, minerals, trace elements and vitamins in a biologically bound form in the following ratio of components, wt.%:

amino acids 80.0-90.0 minerals 3.0-5.0 trace elements 0.005-0.010 vitamins 0.001-0.003

In this case, cartilage is used as animal feed, and the peptide complex that supports the function of cartilage tissue contains amino acids, minerals, trace elements and vitamins in a biologically related form in the following ratio, wt.%:

amino acids 50.0-60.0 minerals 7.0-9.0 trace elements 0.002-0.008 vitamins 0.001-0.003

In this case, the brain is used as animal raw material, and the peptide complex that supports the function of the brain contains amino acids, minerals, trace elements and vitamins in a biologically related form in the following ratio of components, wt.%:

amino acids 50.0-60.0 minerals 4.0-6.0 trace elements 0.002-0.004 vitamins 0.001-0.003

At the same time, thymus is used as animal raw material, and the peptide complex that supports the function of the immune system contains amino acids, minerals, trace elements and vitamins in a biologically related form in the following ratio of components, wt.%:

amino acids 55.0-65.0 minerals 3.0-5.0 trace elements 0.001-0.003 vitamins 0.010-0.030

In this case, the maximum absorption of the target product in the ultraviolet spectrum is (270 ± 5) nm.

The proposed technology, unlike the known ones, allows one to obtain a peptide complex purified from nucleic, protein, and lipid impurities with a molecular mass of its peptide components from 1000 to 12000 Da, containing trace elements, minerals and vitamins in a biologically related form, which has a pronounced tissue-specific activity, which achieved by the proposed sequence of technological operations and the conditions for their implementation, including temperature, time and other characteristics, as well as using Hovhan substances, including feedstock from an extractant and others.

The invention is illustrated in tables.

Table 1 shows the amino acid composition and the ratio of amino acids included in the peptide complex isolated from the liver.

Table 2 shows the mineral content in the peptide complex isolated from the liver.

Table 3 shows the microelement composition of the peptide complex isolated from the liver.

Table 4 presents the results of the content of vitamins in the peptide complex isolated from the liver.

Table 5 presents the chemical composition of the peptide complex isolated from the liver.

Table 6 shows the effect of the peptide complex isolated from the liver on the development of liver explants.

Table 7 shows the amino acid composition and the ratio of amino acids included in the peptide complex isolated from the pancreas.

Table 8 shows the mineral content in the peptide complex isolated from the pancreas.

Table 9 shows the microelement composition of the peptide complex isolated from the pancreas.

Table 10 shows the vitamin content in the peptide complex isolated from the pancreas.

Table 11 shows the chemical composition of the peptide complex isolated from the pancreas.

Table 12 shows the effect of the peptide complex isolated from the pancreas on the development of pancreatic explants.

Table 13 shows the amino acid composition and the ratio of amino acids included in the peptide complex isolated from the thyroid gland.

Table 14 shows the mineral content in the peptide complex isolated from the thyroid gland.

Table 15 shows the microelement composition of the peptide complex isolated from the thyroid gland.

Table 16 presents the vitamin composition of the peptide complex isolated from the thyroid gland.

Table 17 shows the chemical composition of the peptide complex isolated from the thyroid gland.

Table 18 shows the effect of the peptide complex isolated from the thyroid gland on the development of thyroid explants.

Table 19 shows the amino acid composition and the ratio of amino acids included in the peptide complex isolated from blood vessels.

Table 20 shows the mineral composition of the peptide complex isolated from blood vessels.

Table 21 shows the microelement content of the peptide complex isolated from blood vessels.

Table 22 presents the vitamin composition of the peptide complex isolated from blood vessels.

Table 23 shows the chemical composition of the peptide complex isolated from blood vessels.

Table 24 shows the effect of a peptide complex isolated from blood vessels on the development of vascular explants.

Table 25 shows the amino acid composition and the ratio of amino acids included in the peptide complex isolated from cartilage.

Table 26 shows the mineral composition of the peptide complex isolated from cartilage.

Table 27 shows the microelement composition of the peptide complex isolated from cartilage.

Table 28 shows the vitamin composition of the peptide complex isolated from cartilage.

Table 29 shows the chemical composition of the peptide complex isolated from cartilage.

Table 30 shows the effect of the peptide complex isolated from cartilage on the development of cartilage explants.

Table 31 shows the amino acid composition and the ratio of amino acids included in the peptide complex isolated from the brain.

Table 32 shows the mineral composition of the peptide complex isolated from the brain.

Table 33 presents the microelement composition of the peptide complex isolated from the brain.

Table 34 shows the vitamin composition of the peptide complex isolated from the brain.

Table 35 shows the chemical composition of the peptide complex isolated from the brain.

Table 36 shows the effect of a peptide complex isolated from the brain on the development of brain explants.

Table 37 presents the amino acid composition and the ratio of amino acids included in the peptide complex isolated from the thymus.

Table 38 shows the mineral composition of the peptide complex isolated from the thymus.

Table 39 presents the microelement composition of the peptide complex isolated from the thymus.

Table 40 shows the vitamin composition of the peptide complex isolated from the thymus.

Table 41 presents the chemical composition of the peptide complex isolated from the thymus.

Table 42 shows the effect of the peptide complex isolated from thymus on the development of thymus explants.

The following are examples of the preparation of the peptide complex from animal raw materials, which are used in various organs and tissues of animals (example 1 from the liver, example 3 from the pancreas, example 5 from the thyroid gland, example 7 from the vessels, example 9 from cartilage, example 11 - from the brain, example 13 - from the thymus), as well as examples confirming the tissue-specific action of the peptide complex (example 2 - tissue-specific activity of the peptide complex isolated from the liver, example 4 - tissue-specific activity of the peptide complex hexa isolated from the pancreas, example 6 - tissue-specific activity of the peptide complex isolated from the thyroid gland, example 8 - tissue-specific activity of the peptide complex isolated from blood vessels, example 10 - tissue-specific activity of the peptide complex isolated from cartilage, example 12 - tissue-specific activity of the peptide a complex isolated from the brain, example 14 - tissue-specific activity of a peptide complex isolated from the thymus).

Example 1. A method of obtaining a peptide complex isolated from a liver having tissue-specific activity

As animal raw materials, the liver of calves (not older than 12 months of age) or pigs is used.

250 l of a 3% solution of acetic acid are pumped into the extraction reactor and cooled to a temperature of (20 ± 5) ° С, then 50 kg of crushed raw materials are loaded into the reactor with constant stirring. After 30 minutes, add a 1% solution of zinc chloride in a 3% solution of acetic acid (based on 1 kg of raw materials - 10 g of zinc chloride), the suspension is cooled with constant stirring until it reaches a temperature of plus (7-16) ° C. Subsequent mixing is carried out for 1 hour after every 4 hours of sedimentation. The process is carried out for at least 48 hours at a temperature of plus (7-16) ° C.

The separation of the extract from the ballast substances is carried out on a separator at (5000 ± 500) rpm. Acetone is fed to the reactor for precipitation from the measuring device of acetone at the rate of 5 volumes per 1 volume of liver extract. A clarified liver extract is fed into cooled acetone with a thin stream with constant stirring, mixed and the mixture is left until a precipitate forms at a temperature of plus (3-5) ° С for at least 4 hours. The supernatant was removed by decantation. The formed precipitate is washed on a suction filter, on a grid of which a cotton cloth is put, in double volumes of acetone cooled to plus (7-16) ° С, feeding it from the measuring device. The washed precipitate is wiped through a metal sieve with a hole diameter of 1.5 mm and spread in a thin layer into enameled cuvettes, covered with a double layer of cotton cloth and dried at a temperature of plus (18 ± 2) ° С until the odor of acetone is completely removed. During the drying process, the mass is mixed as the top layer dries. The resulting powder is sieved through a metal sieve with a hole diameter of 0.36 mm The dry powder is weighed, a sample is taken and sent for analysis.

The yield of the target product (biologically active peptide complex powder isolated from the liver) is 70 g per 1 kg of feedstock.

The target product (peptide complex) is a cream to dark brown powder and contains biologically active peptide components. The target product is sparingly soluble in water.

For a more detailed description of the biologically active peptide complex obtained by the proposed method, a study was made of its composition, basic physicochemical properties, and specific biological activity.

The molecular weight of the peptide components included in the peptide complex is determined by gel chromatography on Sephadex G-25 and G-50 (Pharmacia, Sweden). To calibrate the 1.6 x 60 cm column, a Peptide Molecuar Weight Kit MS III marker set (Serva, Germany) was used. It was found that the peptide complex includes substances with a molecular mass of not more than 12,000 Da. Using reverse-phase high-performance liquid chromatography in an acetonitrile gradient (Lichrosorb C ₁₈ sorbent, column 2 × 62 mm), it was found that the peptide complex contains; predominantly low molecular weight peptide fractions (from 70 to 90%), and high molecular weight components in the peptide complex are absent.

According to electrophoresis in a 15% polyacrylamide gel, the molecular weight of the peptide components of the peptide complex is from 1000 to 12000 Da.

To determine the authenticity of the peptide complex, a sample of the target product 10 mg is placed in a test tube, dissolved with thorough stirring in 5 ml of water. The solution is filtered through a paper filter. To prepare a biuret reagent, 90 g of potassium-sodium tartrate is dissolved in 400 ml of 0.2 N. sodium hydroxide solution, add 10 g of copper sulfate 5-aqueous, after dissolving add 10 g of potassium iodide and adjust the volume of the solution to 0.2 N. caustic soda solution up to 2 l. 5 ml of biuret reagent is added to the test solution. Staining the solution with violet color indicates the peptide bonds present in the complex. As a comparison substance, water is used.

The identification of the active peptide complex is carried out using ultraviolet spectrophotometry. For this, 10 mg of the peptide complex is dissolved in water in a 100 ml volumetric flask and the volume of the solution is adjusted to the mark with water. An ultraviolet spectrum of the peptide complex is measured on a spectrophotometer in quartz cuvettes with a layer thickness of 10 mm in the wavelength range from 250 to 300 nm. As a comparison solution, water is used. The spectrum should have a pronounced maximum at a wavelength of (270 ± 5) nm. The ratio of optical densities at wavelengths of 275 nm (D ₂₇₅ ) and 260 nm (D ₂₆₀ ) D ₂₇₅ / D ₂₆₀ should be at least 1.0.

To determine the protein in the peptide complex, 25 mg of the product (accurately weighed) is placed in a 100 ml volumetric flask, dissolved in water, mixed with a magnetic stirrer, and the solution volume is adjusted to the mark with water. The solution is filtered through a paper filter. The optical density of the obtained samples is measured in quartz cuvettes with a layer thickness of 10 mm at wavelengths of 280 nm (D ₂₈₀ ) and 260 nm (D ₂₆₀ ). As a comparison solution, water is used.

The protein content (X) in the product in mg / mg is calculated by the formula:

The protein content in the product should be 0.2-0.5 mg / mg.

To determine the content of residual organic solvents (acetone), a solution of a standard sample (RSO) of acetone is prepared. About 10 mg (accurately weighed) of acetone (GOST 2603-79, analytical grade) is placed in a volumetric flask with a capacity of 100 ml, the solution volume is adjusted to the mark with an aqueous ammonia solution and mixed. The solution is used freshly prepared.

To determine the acetone content, about 100 mg (accurately weighed) of the product is dissolved in 5 ml of ammonia solution, the resulting solution is filtered through a paper filter. 2 μl of the resulting solution and a solution of PCO of acetone are alternately chromatographed on a gas analyzer ("Chrom-5" or similar) with a flame ionization detector, obtaining at least 5 chromatograms.

The percentage of acetone (X) in the final product as a percentage is calculated by the formula:

where S ₁ - the average value of the peak areas of acetone from the chromatograms of the test solution;

S ₀ - the average value of the peak areas of acetone from the chromatograms of a solution of OCO acetone;

m ₁ - masa of a sample of a product, in mg;

m ₀ is the weight of the sample of OCO acetone, in mg;

V ₁ - the volume of the test solution, in ml;

V ₀ - the volume of the solution of the OCO acetone, in ml

The results of the analysis are considered reliable if the requirements of the test "Suitability of the chromatographic system" are met.

The presence of a residual amount of acetone of not more than 1% is allowed.

Bacteriological indicators of the peptide complex are analyzed according to GOST 10444.2-94, GOST 10444.15-94, GOST R 50474-93, GOST R 50480-93.

The content of toxic elements in the peptide complex is determined according to GOST 26927-86, GOST 26930-86, GOST 26932-86, GOST 26933-86.

The results of the study of the component composition of the peptide complex isolated from the liver are presented in the following tables.

The amino acid composition of the peptide complex is determined using an LKB-3201 analyzer (Sweden). The analysis showed that the peptide complex isolated from the liver contains the main spectrum of amino acids, including essential amino acids (see Table 1).

Table 1 Name of amino acid The amino acid content, nmol / mg Asp 555 Thr 271 Ser 306 Glu 702 Pro 264 Gly 385 Ala 426 Val 312 Ile 211 Leu 456 Tyr 120 Phe 194 His 138 Lys 333 Arg 225

The amino acid content shown in Table 1 corresponds to the total amount of amino acids in the peptide complex equal to 65-75 wt.%.

The determination of the content of mineral substances in the peptide complex isolated from the liver is carried out using a flame photometer (sodium and potassium), titrometric methods (calcium and magnesium), and calorimetric (phosphorus, iron). In the process of research, it was found that the mineral composition of the peptide complex isolated from the liver is represented by the most important mineral substances for the normal functioning of human organs and systems in calcium, magnesium, iron, phosphorus, as well as potassium and sodium (see Table 2) .

table 2 Product name The content of mineral substances, mcg / 20 mg Sa Mg Fe P TO Na S Peptide complex isolated from the liver 18.01 1.77 2.44 14.35 22.11 490.45 222.87

The mineral content shown in Table 2 corresponds to the total amount of mineral substances in the peptide complex, equal to 3.0-5.0 wt.%.

The content of trace elements in the peptide complex isolated from the liver is determined by emission spectrometry using a spectrochemical system GBC, and are shown in Table 3.

The determination of vitamins in a peptide complex isolated from the liver is carried out by generally accepted methods based on the ability of vitamins to give characteristic color reactions with a number of chemical compounds, the color intensity of which is proportional to the concentration of the vitamin and can be determined photocolometrically, and is shown in Table 4.

Table 3 Product name The content of trace elements, mcg / 20 mg Al Mn Cu With Mo Peptide complex isolated from the liver 0.2158 0.0192 0.4157 0.0027 0.0299

The content of trace elements shown in Table 3 corresponds to the total number of trace elements in the peptide complex, equal to 0.003-0.008 wt.%.

Table 4 Name
product The content of vitamins, mcg / 20 mg In ₁ In ₂ PP FROM BUT E Peptide complex isolated from the liver 0.08 0.059 3,012 traces 0.019 0,040

The vitamin content shown in Table 4 corresponds to the total amount of vitamins in the peptide complex, equal to 0.01-0.03 wt.%.

An analysis of the chemical composition of the target product indicates the absence of carbohydrates in it and its low energy value, which allows it to be used in dietary nutrition (see Table 5).

Table 5 Name
product Energy
cal / 20 mg value The content of components, mg / 20 mg Squirrels Fats Carbohydrates Ash Peptide complex isolated from the liver 42.15 8.12 0.92 traces 0.28

The information on the chemical composition of the product obtained by the proposed method, indicating the presence of physiological concentrations of amino acids, minerals, trace elements and vitamins, demonstrates the chemical composition of the tissue from which the extraction was carried out, and confirms the content of the listed substances in a biologically bound form.

Example 2. Tissue-specific activity of a peptide complex isolated from the liver

To study the tissue-specific activity of the peptide complex isolated by the proposed method from the liver, the effect of the target product on the growth of the organotypic culture of the liver of mature Wistar rats weighing 150-200 g was studied.

The rat liver fragments prepared under sterile conditions were divided into smaller parts of about 1 mm ^{3 in} size, which were placed in Petri dishes with a collagen-coated bottom. The nutrient medium consisted of 35% Eagle medium, 35% Hanks solution, 25% fetal calf serum, 5% chicken fetal extract, 0.6% glucose, 0.5 units / ml insulin, 100 units / ml gentamicin. The studied peptide complex isolated from the liver was introduced into the culture medium at concentrations from 0.01 to 20 ng / ml to reveal its effective concentrations.

In petri dishes with experimental explants, 3 ml of nutrient medium containing the peptide complex isolated from the liver in the studied concentration was added, and in Petri dishes with control explants, 3 ml of nutrient medium; Thus, experimental and control explants developed in equal volumes of culture medium. Petri dishes were placed in a thermostat at a temperature of (37 ± 0.5) ° С and after 3 days they were examined under a phase contrast microscope. The area index (PI) was determined, which was calculated in arbitrary units as the ratio of the area of the entire explant together with the zone of evicted cells to the area of the central zone of the explant.

To visualize the explants, a microfit nozzle was used for a microscope (series 10, MTN-13 Alfa-Telecom, Russia). To calculate the area index of the explants, the Photo M 1.2 program was used. The significance of differences in the area indices of the control and experimental explants was evaluated using Student's t-test. The values of the area index were expressed as a percentage, the control value of IP was taken as 100%.

When using the peptide complex isolated from the liver at concentrations of 0.05, 0.1 and 10 ng / ml, a significant increase in explant PI by 18-28% was observed compared to the control PI values. The data obtained are presented in Table 6.

Table 6 Indicator The concentration of the peptide complex isolated from the liver, ng / ml 0.01 0.05 0.1 0.5 1,0 2.0 10.0 20,0 PI,% in relation to control 5 eighteen* twenty* 22 * fourteen 22 * 28 * eighteen* * p <0.05 compared with the control.

Thus, in relation to liver tissue, the peptide complex isolated from the liver has a tissue-specific effect, which manifests itself in stimulating the growth of explants, which is confirmed in experiments on rats, which allows us to consider its use as an agent supporting liver function.

Example 3. A method of obtaining a peptide complex isolated from the pancreas with tissue-specific activity

As animal raw materials, the pancreas of calves (not older than 12 months of age) or pigs is used.

The selection of the peptide complex from the pancreas is carried out as described in example 1, except that the yield of the target product (powder of a biologically active peptide complex isolated from the pancreas) is 60 g per 1 kg of feedstock.

For a more detailed description of the biologically active peptide complex obtained by the proposed method, a study was made of its composition, basic physicochemical properties, and specific biological activity.

The main characteristics of the peptide complex isolated from the pancreas, determined by the methods described in example 1, are presented in Tables 7-11.

Amino acid analysis showed that the main spectrum of amino acids, including essential amino acids, is present in the peptide complex isolated from the pancreas (see Table 7).

The mineral composition of the peptide complex isolated from the pancreas is represented by the most important mineral substances for the normal functioning of human organs and systems in calcium, magnesium, iron, phosphorus, as well as potassium and sodium, which are shown in Table 8.

The results of the analysis of the microelement composition of the peptide complex isolated by the proposed method from the pancreas are presented in Table 9.

The vitamin composition of the peptide complex isolated by the proposed method from the pancreas is characterized by a significant content of vitamin PP, B ₂ , as well as the presence of vitamins B ₁ , A and E, which is shown in Table 10.

An analysis of the chemical composition of the target product indicates the absence of carbohydrates in it and its low energy value, which allows it to be used in dietary nutrition (see Table 11).

Table 7 Name of amino acid The amino acid content, nmol / mg Asp 388 Thr 171 Ser 212 Glu 505 Pro fifteen) Gly 401 Ala 314 Val 305 Ile 144 Leu 352 Tyr 81 Phe 111 His 129 Lys 306 Arg 187

The amino acid content shown in Table 7 corresponds to the total amount of amino acids in the peptide complex equal to 45-55 wt.%.

Table 8 Name
product The content of mineral substances, mcg / 20 mg Sa Mg Fe P TO Na S Peptide complex isolated from the pancreas 30.14 2.00 1.01 76.14 100.54 1921.41 306.17

The content of mineral substances presented in Table 8 corresponds to the total amount of mineral substances in the peptide complex, equal to 10.0-15 wt.%.

Table 9 Product name The content of trace elements, mcg / 20 mg Al Mn Cu With Mo Peptide complex isolated from the pancreas 0.3211 0,0512 0,0712 0 0.0190

The content of trace elements shown in Table 9 corresponds to the total number of trace elements in the peptide complex, equal to 0.001-0.003 wt.%.

Table 10 Name
product The content of vitamins, mcg / 20 mg B ₁ In ₂ PP FROM BUT E Peptide complex isolated from the pancreas 0.09 0.212 1,255 traces 0.011 0.221

The vitamin content shown in Table 10 corresponds to the total amount of vitamins in the peptide complex, equal to 0.005-0.010 wt.%.

Table 11 Name
product Energy
cal / 20 mg value The content of components, mg / 20 mg Squirrels Fats Carbohydrates Ash Peptide complex isolated from the pancreas 41.12 6.05 1.47 traces 1,0

The information on the chemical composition of the product obtained by the proposed method, indicating the presence of physiological concentrations of amino acids, minerals, trace elements and vitamins, demonstrates the chemical composition of the tissue from which the extraction was carried out, and confirms the content of the listed substances in a biologically bound form.

Example 4. Tissue-specific activity of a peptide complex isolated from the pancreas

To study the tissue-specific activity of the peptide complex isolated from the pancreas by the proposed method, its effect on the growth of the organotypic pancreatic culture of mature rats was studied. The method is described in detail in example 2.

The experiments were carried out on 29 fragments of the pancreas of Wistar rats weighing 150-200 g. Fragments of the pancreas were placed in a nutrient medium and cultured in Petri dishes in an incubator at (37 ± 0.5) ° С for 2 days. A peptide complex isolated from the pancreas was added to the experimental medium at concentrations from 0.01 to 20 ng / ml.

When using a peptide complex isolated from the pancreas at concentrations of 0.05, 0.1 and 10 ng / ml, a significant increase in the explants IP was observed by 16-27% compared with the control IP values. The data obtained are presented in Table 12.

Table 12 Indicator The concentration of the peptide complex isolated from the pancreas, ng / ml 0.01 0.05 0.1 0.5 1,0 2.0 10.0 20,0 PI,% in relation to control 8 twenty* 27 * twenty* 16* 22 * 24 * 16* • p <0.05 compared with the control.

Thus, in relation to pancreatic tissue, the peptide complex isolated from the pancreas has a tissue-specific effect, which manifests itself in stimulating the growth of explants, which is confirmed in experiments on rats, which allows us to consider its use as an agent that supports pancreatic function.

Example 5. A method of obtaining a peptide complex isolated from the thyroid gland with tissue-specific activity

As the animal feed, the thyroid gland of calves (not older than 12 months of age) or pigs is used.

The selection of the peptide complex from the thyroid gland is carried out as described in example 1, except that the yield of the target product (powder of a biologically active peptide complex isolated from the thyroid gland) is 46 g per 1 kg of feedstock.

For a more detailed description of a biologically active peptide complex isolated from the thyroid gland obtained by the proposed method, a study was made of its composition, basic physicochemical properties, and specific biological activity.

The main characteristics of the obtained peptide complex isolated from the thyroid gland, determined by the methods described in example 1, are presented in Tables 13-17.

Amino acid analysis of the target product showed that the main spectrum of amino acids, including essential amino acids, is present in the peptide complex isolated from the thyroid gland (see Table 13).

Table 13 Name of amino acid The amino acid content, nmol / mg Asp 502 Thr 241 Ser 405 Glu 891 Pro 377 Gly 505 Ala 594 Val 418 Ile 158 Leu 588 Tyr 126 Phe 251 His 131 Lys 305 Arg 201

The amino acid content shown in Table 13 corresponds to the total amount of amino acids in the peptide complex equal to 70-80 wt.%.

The mineral composition of the peptide complex isolated from the thyroid gland is represented by the most important mineral substances in the optimal amount for the normal functioning of human organs and systems - calcium, magnesium, iron, phosphorus, as well as potassium and sodium, which are shown in Table 14.

The results of the analysis of the microelement composition of the peptide complex isolated from the thyroid gland are presented in Table 15.

The vitamin composition of the peptide complex isolated from the thyroid gland is characterized by a significant content of vitamin PP, B ₂ , as well as the presence of vitamins B ₁ , A and E, which is shown in Table 16.

An analysis of the chemical composition of the target product indicates the absence of carbohydrates in it and its low energy value, which allows it to be used in dietary nutrition (see Table 17).

Table 14 Name
product The content of mineral substances, mcg / 20 mg Sa Mg Fe P TO Na S Peptide complex isolated from the thyroid gland 20.14 1,02 3.87 42.07 40.11 261.12 277.18

The mineral content shown in Table 14 corresponds to the total amount of mineral substances in the peptide complex, equal to 3.0-5.0 wt.%.

Table 15 Name
product The content of trace elements, mcg / 20 mg Al Mn Cu With Mo Peptide complex isolated from the thyroid gland 0.5080 0.0420 0.0835 0 0.0102

The content of trace elements shown in Table 15 corresponds to the total number of trace elements in the peptide complex equal to 0.002-0.004 wt.%.

Table 16 Name
product The content of vitamins, mcg / 20 mg B ₁ In ₂ PP FROM BUT E Peptide complex isolated from the thyroid gland 0.19 0,020 0,035 traces 0,029 0,057

The vitamin content shown in Table 16 corresponds to the total amount of vitamins in the peptide complex, equal to 0.001-0.003 wt.%.

Table 17 Name
product Energy
value, cal / 20 mg The content of components, mg / 20 mg Squirrels Fats Carbohydrates Ash Peptide complex isolated from the thyroid gland 44.84 7.01 1.33 traces 0.18

The information on the chemical composition of the product obtained by the proposed method, indicating the presence of physiological concentrations of amino acids, minerals, trace elements and vitamins, demonstrates the chemical composition of the tissue from which the extraction was carried out, and confirms the content of the listed substances in a biologically bound form.

Example 6. Tissue-specific activity of a peptide complex isolated from the thyroid gland

To study the tissue-specific activity of the peptide complex isolated by the proposed method from the thyroid gland, we studied the effect of the target product on the growth of the organotypic thyroid culture of adult rats. The method is described in detail in example 2.

The experiments were carried out on 35 fragments of the thyroid gland of Wistar rats weighing 150-200 g. Thyroid fragments were placed in a nutrient medium and cultured in Petri dishes in an incubator at (37 ± 0.5) ° С for 2 days. A peptide complex isolated from the thyroid gland was added to the experimental medium at concentrations from 0.01 to 20 ng / ml.

When using the peptide complex isolated from the thyroid gland in concentrations of 0.05, 0.1 and 10 ng / ml, a significant increase in the explant PI by 18-24% was observed compared with the control PI values.

Table 18 shows the effect of the peptide complex isolated from the thyroid gland on the development of thyroid explants.

Table 18 Indicator The concentration of the peptide complex isolated from the thyroid gland, ng / ml 0.01 0.05 0.1 0.5 1,0 2.0 10.0 20,0 PI,% in relation to control 7 eighteen* twenty* twenty* fifteen twenty* 24 * twenty* • p <0.05 compared with the control.

Thus, in relation to thyroid tissue, the peptide complex isolated from the thyroid gland has a tissue-specific effect, which is manifested in stimulating the growth of explants, which is confirmed in experiments on rats, which allows us to consider its use as an agent supporting the thyroid function.

Example 7. A method of obtaining a peptide complex isolated from blood vessels with tissue-specific activity

The vessels of calves (not older than 12 months of age) or pigs are used as animal raw materials.

The selection of the peptide complex from the vessels is carried out as described in example 1, except that the yield of the target product (powder of the biologically active peptide complex isolated from the vessels) is 28 g per 1 kg of feedstock.

For a more detailed description of the peptide complex obtained by the proposed method, a study was made of its composition, basic physicochemical properties, and specific biological activity.

The main characteristics of the obtained peptide complex isolated from blood vessels, determined by the methods described in example 1, are presented in Tables 19-23. Amino acid analysis of the target product showed that the main spectrum of amino acids, including essential amino acids, is present in the peptide complex isolated from blood vessels (see Table 19).

Table 19 Name of amino acid The amino acid content, nmol / mg Asp 755 Thr 361 Ser 402 Glu 999 Pro 374 Gly 452 Ala 502 Val 401 Ile 247 Leu 504 Tyr 200 Phe 238 His 231 Lys 498 Arg 356

The amino acid content shown in Table 19 corresponds to the total amount of amino acids in the peptide complex equal to 80.0-90.0 wt.%.

The mineral composition of the peptide complex isolated from blood vessels is represented by the most important mineral substances in the optimal amount for the normal functioning of human organs and systems - calcium, magnesium, iron, phosphorus, as well as potassium and sodium, which are shown in Table 20.

Table 20 Name
product The content of mineral substances, mcg / 20 mg Sa Mg Fe P TO Na S Peptide complex isolated from blood vessels 25.74 3.03 1.71 25.17 34.55 222.51 444.45

The content of mineral substances shown in Table 20 corresponds to the total amount of mineral substances in the peptide complex, equal to 3.0-5.0 wt.%. The results of the analysis of the microelement composition of the peptide complex isolated from blood vessels are presented in Table 21.

Table 21 Name
product The content of trace elements, mcg / 20 mg Al Mn Cu With Mo Peptide complex isolated from blood vessels 0.9812 0.0123 0.5846 0 0.0110

The content of trace elements shown in Table 21 corresponds to the total number of trace elements in the peptide complex equal to 0.005-0.010 wt.%. The vitamin composition of the peptide complex isolated from blood vessels is characterized by a significant content of vitamin PP, B ₂ , as well as the presence of vitamins B ₁ , A and E, which are presented in Table 22.

Table 22 Name
product The content of vitamins, mcg / 20 mg In ₁ In ₂ PP FROM BUT E Peptide complex isolated from blood vessels 0.33 0.002 0,041 traces 0,023 0,039

The vitamin content shown in Table 22 corresponds to the total amount of vitamins in the peptide complex equal to 0.001-0.003 wt.%.

An analysis of the chemical composition of the target product indicates the absence of carbohydrates in it and its low energy value, which allows it to be used in dietary nutrition (see Table 23).

Table 23 Name
the drug Energy The content of components, mg / 20 mg cal / 20 mg value Squirrels Fats Carbohydrates Ash Peptide complex isolated from blood vessels 39.15 7.85 0.86 traces 0.35

The information on the chemical composition of the product obtained by the proposed method, indicating the presence of physiological concentrations of amino acids, minerals, trace elements and vitamins, demonstrates the chemical composition of the tissue from which the extraction was carried out, and confirms the content of the listed substances in a biologically bound form.

Example 8. Tissue-specific activity of a peptide complex isolated from blood vessels

To study the tissue-specific activity of the peptide complex isolated from the vessels of the proposed method, the effect of the target product on the growth of the organotypic culture of vessels of mature rats was investigated. The method is described in detail in example 2.

The experiments were performed on 24 fragments of vessel tissue of the Wistar rats weighing 150-200 g. Vascular tissue fragments were placed in a nutrient medium and cultured in Petri dishes in an incubator at (37 ± 0.5) ° С for 2 days. A peptide complex isolated from blood vessels was added to the experimental medium at concentrations from 0.01 to 20 ng / ml.

When using a peptide complex isolated from blood vessels at concentrations of 0.05, 0.1 and 10 ng / ml, a significant increase in the explant PI by 18–25% was observed compared with the reference PI values.

Table 24 shows the effect of a peptide complex isolated from blood vessels on the development of vascular explants.

Table 24 Indicator The concentration of the peptide complex isolated from blood vessels, ng / ml 0.01 0.05 0.1 0.5 1,0 2.0 10.0 20,0 PI,% in relation to control 7 22 * 24 * 16 fourteen eighteen* 25 * fourteen • p <0.05 compared with the control.

Thus, in relation to vascular tissue, the peptide complex isolated from the vessels exerts a tissue-specific effect, which manifests itself in stimulating the growth of explants, which is confirmed in experiments on rats, which allows us to consider its use as an agent that supports vascular function.

Example 9. A method of obtaining a peptide complex isolated from cartilage with tissue-specific activity

Cartilage of calves (not older than 12 months of age) or pigs is used as animal feed.

The selection of the peptide complex from cartilage is carried out as described in example 1, except that the yield of the final product (biologically active peptide complex powder isolated from cartilage) is 30 g per 1 kg of feedstock.

For a more detailed description of a biologically active peptide complex obtained from cartilage by the proposed method, a study was made of its composition, basic physicochemical properties, and specific biological activity.

The main characteristics of the obtained peptide complex isolated from cartilage, determined by the methods described in example 1, are presented in Tables 25-29.

Amino acid analysis of the target product showed that in the peptide complex isolated from cartilage, there is a basic spectrum of amino acids, including essential amino acids (see Table 25).

Table 25 Name of amino acid The amino acid content, nmol / mg Asp 481 Thr 198 Ser 244 Glu 602 Pro 201 Gly 416 Ala 402 Val 233 Ile 151 Leu 375 Tyr 111 Phe 154 His 120 Lys 305 Arg 198

The amino acid content shown in Table 25 corresponds to the total amount of amino acids in the peptide complex equal to 50-60 wt.%.

The mineral composition of the peptide complex isolated from cartilage is represented by the most important mineral substances in the optimal amount for the normal functioning of human organs and systems - calcium, magnesium, iron, phosphorus, as well as potassium and sodium, as shown in Table 26.

Table 26 Name
product The content of mineral substances, mcg / 20 mg Sa Mg Fe P TO Na S Peptide complex isolated from cartilage 21.17 2.05 1.33 28.89 90.36 1222.44 270.62

The content of mineral substances shown in Table 26 corresponds to the total amount of mineral substances in the peptide complex, equal to 7.0-9.0 wt.%.

The results of the analysis of the microelement composition of the peptide complex isolated from cartilage are presented in Table 27.

The vitamin composition of the peptide complex isolated from cartilage is characterized by a significant content of vitamin PP, B ₂ , as well as the presence of vitamins B ₁ , A and E, which are presented in Table 28.

Table 27 Product name The content of trace elements, mcg / 20 mg Al Mn Cu With Mo Peptide complex isolated from cartilage 0.5926 0.0198 0,0702 0.0010 0.0111

The content of trace elements shown in Table 27 corresponds to the total number of trace elements in the peptide complex equal to 0.002-0.008 wt.%.

Table 28 Name
product The content of vitamins, mcg / 20 mg B ₁ In ₂ PP FROM BUT E Peptide complex isolated from cartilage 0.34 0.007 0.014 traces 0,020 0,040

The vitamin content shown in Table 28 corresponds to the total amount of vitamins in the peptide complex, equal to 0.001-0.003 wt.%.

An analysis of the chemical composition of the target product indicates the absence of carbohydrates in it and its low energy value, which allows it to be used in diet food (see Table 29).

Table 29 Name
product Energy The content of components, mg / 20 mg cal / 20 mg value Squirrels Fats Carbohydrates Ash Peptide complex isolated from cartilage 42.63 6.94 1,54 traces 0.48

The information on the chemical composition of the product obtained by the proposed method, indicating the presence of physiological concentrations of amino acids, minerals, trace elements and vitamins, demonstrates the chemical composition of the tissue from which the extraction was carried out, and confirms the content of the listed substances in a biologically bound form.

Example 10. Tissue-specific activity of a peptide complex isolated from cartilage

To study the tissue-specific activity of the peptide complex isolated from the cartilage by the proposed method, the effect of the target product on the growth of the organotypic culture of the cartilage tissue of mature rats was studied. The method is described in detail in example 2.

The experiments were carried out on 30 fragments of cartilage tissue of Wistar rats weighing 150-200 g. Cartilage tissue fragments were placed in a nutrient medium and cultured in Petri dishes in an incubator at (37 ± 0.5) ° С for 2 days. A peptide complex isolated from cartilage in concentrations from 0.01 to 20 ng / ml was added to the experimental medium.

When using a peptide complex isolated from cartilage at concentrations of 0.05, 0.1 and 10 ng / ml. there was a significant increase in explant PI by 19-25% compared with the control PI values.

Table 30 shows the effect of the peptide complex isolated from cartilage on the development of cartilage tissue explants.

Table 30 Indicator The concentration of the peptide complex of cartilage, ng / ml 0.01 0.05 0.1 0.5 1,0 2.0 10.0 20,0 PI,% in relation to control 5 twenty* 22 * 19* 16 twenty* 25 * 12 • p <0.05 compared with the control.

Thus, in relation to cartilage tissue, the peptide complex isolated from cartilage has a tissue-specific effect, manifested in stimulating the growth of explants, which is confirmed in experiments on rats, which allows us to consider its use as an agent supporting the function of cartilage tissue.

Example 11. A method of obtaining a peptide complex isolated from the brain with tissue-specific activity

As animal raw materials, the brains of calves (not older than 12 months of age) or pigs are used.

The selection of the peptide complex from the brain is carried out as described in example 1, except that the yield of the final product (powder of a biologically active peptide complex isolated from the brain) is 44 g per 1 kg of feedstock.

For a more detailed description of a biologically active peptide complex obtained from the brain by the proposed method, a study was made of its composition, basic physical and chemical properties, and specific biological activity.

The main characteristics of the obtained peptide complex isolated from the brain, determined by the methods described in example 1, are presented in Tables 31-35.

Amino acid analysis of the target product showed that the main spectrum of amino acids, including essential amino acids, is present in the peptide complex isolated from the brain (see Table 31).

Table 31 Name of amino acid The amino acid content, nmol / mg / Asp 491 Thr 198 Ser 285 Glu 602 Pro 191 Gly 305 Ala 355 Val 233 Ile 164 Leu 384 Tyr 101 Phe 177 His 122 Lys 255 Arg 197

The amino acid content shown in Table 31 corresponds to the total amount of amino acids in the peptide complex equal to 50-60 wt.%.

The mineral composition of the peptide complex isolated from the brain is represented by the most important mineral substances in the optimal amount for the normal functioning of human organs and systems - calcium, magnesium, iron, phosphorus, as well as potassium and sodium, which are shown in Table 32.

Table 32 Name
product The content of mineral substances, mcg / 20 mg Sa Mg Fe P TO Na S Peptide complex isolated from the brain 36.11 2.88 1.00 50,44 21.18 674.25 147.30

The mineral content shown in Table 32 corresponds to the total amount of mineral substances in the peptide complex, equal to 4.0-6.0 wt.%. The results of the analysis of the microelement composition of the peptide complex isolated from the brain are presented in Table 33.

Table 33 Name
product The content of trace elements, mcg / 20 mg Al Mn Cu With Mo Peptide complex isolated from the brain 0.2844 0.0131 0.1998 0.0028 0.0190

The content of trace elements shown in Table 33 corresponds to the total number of trace elements in the peptide complex, equal to 0.002-0.004 wt.%.

The vitamin composition of the peptide complex isolated from the brain is characterized by a significant content of vitamin PP, B ₂ , as well as the presence of vitamins B ₁ , A and E, which are presented in Table 34.

Table 34 Name
product The content of vitamins, mcg / 20 mg B ₁ In ₂ PP C BUT E Peptide complex isolated from the brain 0,07 0,027 0,063 traces 0.010 0.005

The vitamin content shown in Table 34 corresponds to the total amount of vitamins in the peptide complex, equal to 0.001-0.003 wt.%.

An analysis of the chemical composition of the target product indicates the absence of carbohydrates in it and its low energy value, which allows it to be used in dietary nutrition (see Table 35).

Table 35 Name
the drug Energy The content of components, mg / 20 mg cal / 20 mg value Squirrels Fats Carbohydrates Ash Peptide complex isolated from the brain 48.13 6.42 1.98 traces 0.41

The information on the chemical composition of the product obtained by the proposed method, indicating the presence of physiological concentrations of amino acids, minerals, trace elements and vitamins, demonstrates the chemical composition of the tissue from which the extraction was carried out, and confirms the content of the listed substances in a biologically bound form.

Example 12. Tissue-specific activity of a peptide complex isolated from the brain

To study the tissue-specific activity of the peptide complex isolated by the proposed method from the brain, the effect of the target product on the growth of the organotypic brain culture of adult rats was studied. The method is described in detail in example 2.

The experiments were carried out on 32 fragments of the brain of Wistar rats weighing 150-200 g. Fragments of the rat brain were placed in a nutrient medium and cultured in Petri dishes in an incubator at (37 ± 0.5) ° С for 2 days. A peptide complex isolated from the brain was added to the experimental medium at concentrations from 0.01 to 20 ng / ml.

When using a peptide complex isolated from the brain at concentrations of 0.05, 0.1 and 10 ng / ml, a significant increase in explant PI by 20-28% was observed compared with the reference PI values.

Table 36 shows the effect of a peptide complex isolated from the brain on the development of brain explants.

Table 36 Indicator The concentration of the peptide complex isolated from the brain, ng / ml 0.01 0.05 0.1 0.5 1,0 2.0 10.0 20,0 PI,% in relation to control 8 24 * 24 * 24 * twenty* 26 * 28 * twenty* • p <0.05 compared with the control.

Thus, in relation to brain tissue, a peptide complex isolated from the brain has a tissue-specific effect, manifested in stimulating the growth of explants, which is confirmed in experiments on rats, which allows us to consider its use as an agent supporting brain function.

Example 13. A method of obtaining a peptide complex isolated from a thymus with tissue-specific activity

Thymus (goiter glands) of calves (not older than 12 months of age) or pigs are used as animal raw materials.

The selection of the peptide complex from the thymus is carried out as described in example 1, except that the yield of the final product (biologically active peptide complex powder isolated from the thymus) is 54 g per 1 kg of feedstock.

For a more detailed description of the biologically active peptide complex obtained by the proposed method, a study was made of its composition, basic physicochemical properties, and specific biological activity.

The main characteristics of the obtained peptide complex isolated from the thymus determined by the methods described in example 1 are presented in Tables 37-41.

Amino acid analysis of the target product showed that the main spectrum of amino acids, including essential amino acids, is present in the peptide complex isolated from the thymus (see Table 37).

The mineral composition of the peptide complex isolated from the thymus is represented by the most important mineral substances in the optimal amount for the normal functioning of human organs and systems - calcium, magnesium, iron, phosphorus, as well as potassium and sodium, which are presented in Table 38.

The results of the analysis of the microelement composition of the peptide complex isolated from the thymus are presented in Table 39.

The vitamin composition of the peptide complex isolated from the thymus is characterized by a significant content of vitamins PP, B ₁ as well as the presence of vitamins B ₂ , A and E (see Table 40).

Table 37 Name of amino acid The amino acid content, nmol / mg Asp 398 Thr 241 Ser 288 Glu 605 Pro 233 Gly 477 Ala 451 Val 245 Ile 182 Leu 341 Tyr one hundred Phe 119 His 132 Lys 422 Arg 356

The amino acid content shown in Table 37 corresponds to the total amount of amino acids in the peptide complex equal to 55-65 wt.%.

Table 38 Name
product The content of mineral substances, mcg / 20 mg Sa Mg Fe P TO Na S Peptide complex isolated from thymus 45.19 4.01 4.22 71.61 21.84 465.11 133.48

The mineral content shown in Table 38 corresponds to the total amount of mineral substances in the peptide complex, equal to 3.0-5.0 wt.%.

Table 39 Name
product The content of trace elements, mcg / 20 mg Al Mn Cu With Mo Peptide complex isolated from thymus 0.2459 0.0074 0,0399 0.0030 0.0036

The content of trace elements shown in Table 39 corresponds to the total number of trace elements in the peptide complex, equal to 0.001-0.003 wt.%.

Table 40 Name
product The content of vitamins, mcg / 20 mg B ₁ In ₂ PP FROM BUT E Peptide complex isolated from thymus 0.09 0.007 1,788 traces 0,022 1,000

The vitamin content shown in Table 40 corresponds to the total amount of vitamins in the peptide complex equal to 0.010-0.030 wt.%.

An analysis of the chemical composition of the target product indicates the absence of carbohydrates in it and its low energy value, which allows it to be used in dietary nutrition (see Table 41).

Table 41 Name
product Energy The content of components, mg / 20 mg cal / 20 mg value Squirrels Fats Carbohydrates Ash Peptide complex isolated from thymus 39.65 6.44 1.22 traces 1,02

The information on the chemical composition of the product obtained by the proposed method, indicating the presence of physiological concentrations of amino acids, minerals, trace elements and vitamins, demonstrates the chemical composition of the tissue from which the extraction was carried out, and confirms the content of the listed substances in a biologically bound form.

Example 14. Tissue-specific activity of a peptide complex isolated from thymus

To study the tissue-specific activity of the peptide complex isolated by the proposed method from the thymus, the effect of the target product on the growth of the organotypic thymus culture of mature rats was studied. The method is described in detail in example 2.

The experiments were performed on 32 fragments of the thymus of the Wistar rats weighing 150-200 g. The fragments of goiter were placed in a nutrient medium and cultured in Petri dishes in an incubator at (37 ± 0.5) ° С for 2 days. A peptide complex isolated from the thymus was added to the experimental medium at concentrations from 0.01 to 20 ng / ml.

When using the peptide complex isolated from the thymus at concentrations of 0.05, 0.1 and 10 ng / ml, a significant increase in the explant PI by 20-30% was observed, compared with the reference PI values.

Table 42 shows the effect of the peptide complex isolated from thymus on the development of thymus explants.

Table 42 Indicator Thymus peptide complex concentration, ng / ml 0.01 0.05 0.1 0.5 1,0 2.0 10.0 20,0 PI,% in relation to control 7 26 * 26 * 27 * twenty* 27 * thirty* 16 • p <0.05 compared with the control.

Thus, with respect to the thymus tissue, the peptide complex isolated from the thymus has a tissue-specific effect, which is manifested in stimulating the growth of explants, which is confirmed in experiments on rats, which allows us to consider its use as a means for correcting the functions of the immune system.

The above examples convincingly confirm that the obtained target product is a peptide complex with a molecular weight of peptide components from 1000 to 12000 Da, containing in a biologically related form amino acids, trace elements, minerals and vitamins specific to organs and tissues, of which it is highlighted.

Claims (9)

1. The method of obtaining funds for maintenance therapy with tissue-specific activity, characterized in that the organs of calves not older than 12 months of age or pigs are crushed, add a 3% solution of acetic acid at 20 ± 5 ° C, the extraction is carried out with constant stirring, after 30 min, add a 1% solution of zinc chloride, cool with constant stirring to 7-16 ° C, then mix for 1 hour after every 4 hours of settling for 48 hours, the extract is separated from the ballast substances by separation, add to the extract and the ketone is in a volume ratio of 1: 5, kept at 3-5 ° C for 4 h, the precipitate formed is washed with a double volume of acetone cooled to 7-16 ° C, the washed precipitate is wiped through a metal sieve, the obtained target product, which is a peptide complex with the content of low molecular weight peptide fraction from 70 to 90%, dried at 18 ± 2 ° C. 2. The method according to claim 1, characterized in that the maximum absorption of the target product in the ultraviolet spectrum is 270 ± 5 nm. 3. An agent for maintenance therapy with tissue-specific activity, characterized in that it is obtained by the method according to claim 1 from liver tissue and is a peptide complex that supports liver function, with a molecular weight of the peptide components included in it ranging from 1000 to 12000 Yes , which contains amino acids, minerals, trace elements and vitamins in biologically related form with the following components: Amino acids 120-702 nmol / mg Minerals 1.77-490.45 mcg / 20 mg Trace elements 0.0027-0.4157 mcg / 20 mg Vitamins 0.019-3.012 mcg / 20 mg 4. An agent for maintenance therapy with tissue-specific activity, characterized in that it is obtained by the method according to claim 1 from pancreatic tissue and is a peptide complex with a molecular weight of its peptide components in the range from 1000 to 12000 Yes, supporting pancreatic function gland, contains amino acids, minerals, trace elements and vitamins in biologically related form with the following components: Amino acids 81-505 nmol / mg Minerals 1.01-1921.41 mcg / 20 mg Trace elements 0.0190-0.3211 mcg / 20 mg Vitamins 0.011-1.255 mcg / 20 mg 5. An agent for maintenance therapy, having tissue-specific activity, characterized in that it is obtained by the method according to claim 1 from thyroid tissue and is a peptide complex with a molecular weight of its peptide components ranging from 1000 to 12000 Yes, supporting thyroid function gland, contains amino acids, minerals, trace elements and vitamins in biologically related form with the following components: Amino acids 126-891 nmol / mg Minerals 1.02-277.18 mcg / 20 mg Trace elements 0.0102-0.5080 mcg / 20 mg Vitamins 0.02-0.19 nmol / mg 6. An agent for maintenance therapy with tissue-specific activity, characterized in that it is obtained by the method according to claim 1 from blood vessels and is a peptide complex with a molecular weight of its peptide components ranging from 1000 to 12000 Yes, which supports the function of blood vessels amino acids, trace elements and vitamins in biologically bound form with the following components: Amino acids 200-999 nmol / mg Minerals 1.71-444.45 mcg / 20 mg Trace elements 0.011-0.9812 mcg / 20 mg Vitamins 0.002-0.33 mcg / 20 mg 7. An agent for maintenance therapy, having tissue-specific activity, characterized in that it is obtained by the method according to claim 1 from cartilage tissue and is a peptide complex with a molecular weight of its peptide components ranging from 1000 to 12000 Yes, supporting the function of cartilage tissue , contains amino acids, minerals, trace elements and vitamins in biologically bound form with the following components: Amino acids 111-602 nmol / mg Minerals 1.33-1222.44 mcg / 20 mg Trace elements 0.0010-0.5926 mcg / 20 mg Vitamins 0.007-0.34 mcg / 20 mg 8. An agent for maintenance therapy with tissue-specific activity, characterized in that it is obtained by the method according to claim 1 from brain tissue and is a peptide complex with a molecular weight of its peptide components ranging from 1000 to 12000 Yes, supporting the function of the brain the brain contains amino acids, minerals, trace elements and vitamins in biologically related form with the following components: Amino acids 101-602 nmol / mg Minerals 1.0-674.25 mcg / 20 mg Trace elements 0.0028-0.2844 mcg / 20 mg Vitamins 0.005-0.07 mcg / 20 mg 9. An agent for maintenance therapy, having tissue-specific activity, characterized in that it is obtained by the method according to claim 1 from the thymus and is a peptide complex with a molecular weight of its peptide components in the range from 1000 to 12000 Yes, supporting the function of the immune system, contains amino acids, minerals, trace elements and vitamins in biologically bound form with the following components: Amino acids 100-605 nmol / mg Minerals 4.01-465.11 mcg / 20 mg Trace elements 0.0030-0.2459 mcg / 20 mg Vitamins 0.007-1.788 mcg / 20 mg