RU2041715C1 - Biologically active remedy, method for its producing, preparation containing the mentioned remedy and method for applying the preparation - Google Patents

Abstract

FIELD: medicine. SUBSTANCE: remedy based on water-soluble hydrolysis products represented by certain organic compounds has thermoresistant low molecular weight morphoplasm and cell glycocalyx components modified in embryogenesis and/or proliferation and/or pathologic processes preceding or accompanying the tissue restoration and/or repair present among hydrolysis products. Method for producing the biologically active remedy involves comminuting washed animal tissue raw materials, hydrolyzing the obtained ingredients, their filtering and separating supernatant as the end product. Animal tissue with modified morphoplasm and cell glycocalyx components is used as raw materials homogenized and deprived of undamaged animal tissue elements with supernatant, containing the ingredients with modified morphoplasm and cell glycocalyx components, having been separated before being exposed to hydrolysis. The preparation for normalizing functional state of human or animal organism has biologically active remedy produced on the base of animal tissue elements hydrolysis products and filler thermoresistant low molecular weight morphoplasm and cell glycocalyx components in the amount of 0.001- 85.0% by mass with the rest completed by the filler. The method for normalizing physiological state of human or animal organism involves administering the preparation based on the animal tissue elements hydrolysis products being a composition of the biologically active remedy containing the modified morphoplasm and cell glycocalyx components among hydrolysis products and filler in the amount of 0.001-85.0 and 15.0-99.999% by mass, respectively. EFFECT: enhanced immunomodulating effectiveness. 31 cl, 12 tbl

Description

 The group of inventions relates to the pharmaceutical industry, in particular to the field of obtaining biologically active agents with immunomodulatory effects, for use in medicine and veterinary medicine with the goal of normalizing the physiological state of the human and animal body, in particular, to increase the resistance and productivity of animals.

 The biologically active agent in accordance with the present invention relates to substances of animal origin and includes organic compounds.

 Various pathological disorders in the human and animal body cause deviations from the norm of the parameters of the physiological state. In these cases, the therapeutic effect on the body, including the preventive effect, consists in stabilization, that is, maintaining these parameters in the norm or (if they deviate from the norm) in such an effect on the body in which these parameters are normalized.

 The group of inventions relates to a biologically active agent, a method for its preparation and a preparation based on a biologically active agent used to normalize the physiological state of the body.

 Normalization of the physiological state of the human and animal organisms can be carried out in two ways: firstly, by directly affecting the pathology and, secondly, by indirectly affecting the systems, organs and tissues of the body, which are responsible for maintaining normal parameters of homeostasis.

 The introduction of hormonal drugs, mediators, growth factors, chemotherapy, and the like, provides a direct effect on target cells through reception. For example, in the treatment of diabetes, insulin is administered to supplement the functional deficiency of the pancreas; for violations of the functional activity of the nervous system, neurotransmitters are introduced, which contribute to the normalization of the threshold sensitivity of neurons and, accordingly, to the improvement of transmission of nerve impulses; in cases of impaired functional activity of the body’s immune system, immuno-mediators are used that correlate the immune response, and so on.

 With an indirect effect on the body, the treatment of diseases is provided by mobilizing the internal resources of the body. So, during the processes associated with the regeneration of organs and tissues, with their rehabilitation or restoration of their functions, as well as in inflammatory and infectious processes, they stimulate the body's immune system. Due to this, they provide an improvement in the immunological and, as a consequence, the general condition of the body.

 A biologically active agent based on germline tissue is known, having an immunomodulating property, restoring the physiological state of the body and regulating cell metabolism (FR-A-2413912). The specified biologically active agent (hereinafter referred to as BPS) is made on the basis of the ingredients of crushed and homogenized animal tissue isolated from the embryo, mixed with physiological serum.

 Due to the presence of an immunomodulating factor in the composition of this agent, it has a relatively high immunomodulating property, due to which its use ensures normalization of the physiological state of the body, including treatment, and an increase in animal productivity by 5-15% compared with the control group.

 The effectiveness of the known ALS is still insufficient due to the fact that, along with immunomodulating factors, it contains immunosuppressants that prevent the full manifestation of the immunomodulating effect. The immunomodulatory property of this agent, in fact, is the total result, which is ensured by the simultaneous presence of immunostimulating factors and depressants in it. Thus, in fact, the specific activity of known ALS, depending on the ratio of concentrations of immunostimulating factors and depressants, is relatively small due to the significant concentration of depressants in its composition.

 Another disadvantage of this ALS is the increased content of high molecular weight compounds that are immunogens based on it, leading to the appearance of adverse immune reactions (allergies, delayed hypersensitivity, and the like).

 To a large extent, the disadvantages of the above ALS depend on the method of its preparation, in which the technological process ends with the operation of grinding and homogenizing the raw material from animal tissue, which leads to the mandatory presence of a significant amount of depressants and high molecular weight compounds in its composition.

 BAS is also known, having an immunomodulating property, for normalizing the physiological state of the body, made on the basis of the ingredients of crushed and homogenized tissue, isolated in the form of an extract of the germinal tissues of animal organs (EP-A-0249563).

 The method of its production consists in grinding and homogenizing the raw material from animal tissue, the subsequent removal of insoluble compounds and the use of extracted solutes as the target product (EP-A-0249563).

 The specific activity of the agents obtained by the described method, determined by the resulting ratio of the effects of immunostimulation and immuno-inhibition, is also insufficient. This is because in the method for producing the known BAS in the form of an extract, only soluble proteins are used, and structural proteins are removed, which significantly reduces the content of immunostimulating factors in the composition of the agent.

 In addition, the known ALS also contains high molecular weight compounds that cause adverse immune reactions of the body.

 BAS is also known based on the ingredients of crushed and homogenized animal tissue, obtained in the form of an extract of thermostable tissue components of immunocompetent organs (thymus, spleen, lymph nodes) or hormone-producing tissues (placenta) to normalize the physiological state of the body, including to increase the resistance and productivity of farm animals (SU-A-1442064).

 Known ALS is designed to regulate the immune T-system, and practically does not cause adverse immune reactions when it is used (allergies, delayed hypersensitivity, and so on). Its specific activity in comparison with the already considered ALS is higher due to the higher content of immunostimulating factors.

 However, the use of this tool to normalize the physiological state of the body is limited due to its mediator and / or hormonal nature, which does not allow overdoses due to the danger of disturbing the parameters of the body's homeostasis.

 It is safer to use BAS to normalize the physiological state of the body, obtained in the form of an extract of thermostable components of crushed animal tissue, in which mediator or hormone-producing activity is not so significant (SU-A-139404). This ALS has the same advantages as previously considered ALS, however, it does not exhibit mediator and / or hormonal activity, although it somewhat loses in its immunomodulating activity. These properties of the known BAS are related to the method of its preparation, which consists in grinding animal tissue, its double heat treatment and packaging under aseptic conditions (SU-A-139404).

 Another UAS with an immunomodulatory property, to normalize the physiological state of the body, including for healing injured areas of the body, is based on water-soluble products of hydrolysis of ingredients of crushed raw material from animal tissue, preferably taken from underdeveloped animals (US-A-4455302). At the same time, it includes mineral substances, soluble organic compounds, such as polypeptides, amino acids and mineral substances obtained as a result of tissue hydrolysis.

 This tool is highly effective in relation to normalizing the parameters of the physiological state of the body. This is because the known ALS is characterized by a high ratio of the effects of immunostimulation and immuno-inhibition due to the higher content of immunostimulating factors in its composition.

 However, the effectiveness of the described ALS is still insufficient, due to the content in its composition of a relatively large number of ballast substances and immunosuppressants. In addition, the known ALS can also cause side effects that occur during its use (allergies and others). A disadvantage of the known BAS is that the composition of the target product obtained is inconsistent and varies from pariah to batch, which makes standardization difficult.

 The properties of the known ALS, primarily its immunomodulatory activity, are directly related to the method of its preparation.

In the known method for producing BAS, having an immunomodulating property, to normalize the physiological state of the body, hydrolysis of the ingredients of the washed ground raw material from animal tissue is carried out, followed by centrifugation, filtration of the hydrolysis products and isolation of the target product in the form of a powder or gel. The hydrolysis is carried out with a dilute solution of a weak organic acid at pH 3,6-6,5 and at a temperature of about 68 ^{° C,} and used as a raw material the legs immature poultry no more than 9 weeks of age (US-A- 4455302).

 The raw materials used, as well as the lack of operations before hydrolysis to homogenize and purify undestructed tissue elements with the release of a supernatant containing ingredients with morphoplasma and glycocalyx components of the cells, does not allow to obtain the final product with a high (more than 30%) concentration of these components, and also lower the content of immunosuppressants, neutral and ballast compounds in it.

 A medicament is known for normalizing the physiological state of a human and animal organism based on the products of hydrolysis of animal tissue ingredients represented by organic compounds, which are applied to the affected areas of the body (US-A-4455302).

 In addition to these shortcomings, the disadvantage of the known drug based on ALS is that stimulation of the body's immune system is possible only by local application to the injured areas of the body. Therefore, the effectiveness of such a drug is relatively small.

 These disadvantages are partially eliminated in the known preparation having an immunomodulatory effect to normalize the physiological state of the human and animal body, containing the active principle, which is a biologically active agent based on the products of hydrolysis of animal tissue ingredients, and a filler that is introduced into the body by intramuscular injection in a dose 0.75-1.0 ml / kg body weight with an interval of 3 days and, thus, is used non-locally (SU-A-904707).

 A disadvantage of the known method of using the drug is that it is injected into the body only, as well as the fact that its use causes side effects due to the presence of immunogens that cause allergies, delayed-type hypersensitivity and more.

 The use of raw animal tissue containing a relatively large number of immunosuppressants, and the inability to improve the composition of ALS, on the basis of which the drug is performed using operations performed in the known method, significantly reduces the effectiveness of the known ALS and a drug based on it.

 A significant disadvantage of the known drug is that to ensure sufficient effectiveness, it contains a large number of various biologically active substances, which on the one hand causes the risk of overdose, the occurrence of side effects, and on the other increases the cost of the drug.

 A known method of normalizing the physiological state of humans and animals in diseases caused mainly by pathologically altered and / or foreign cells, microorganisms and / or products of their vital activity, which provides for the introduction into the body of a drug based on the products of hydrolysis of animal tissue ingredients (SU-A-904707).

 The known method is ineffective due to the presence of the disadvantages of the drug described above, including the fact that its use requires the introduction of large doses containing up to 10 mg of biologically active agent per 1 kg of body weight, which causes the risk of overdose and side effects.

 The above disadvantages are eliminated in the proposed biologically active agent having an immunomodulating property, in the proposed method for producing this agent and a preparation based on this agent, as well as in the method of its use for normalizing the physiological state of the body.

 The basis of the invention is the creation of a biologically active agent with an immunomodulating property, a method for its preparation and a drug based on this agent, which would provide high biological activity without manifestations of toxicity and various side effects, as well as the possibility of obtaining the target product of a given composition and with desired properties .

 This is due to the fact that a biologically active agent with an immunomodulating property, based on the water-soluble products of hydrolysis of ingredients of ground animal tissue represented by organic compounds, contains thermostable, low molecular weight components of morphoplasm and glycocalyx of cells modified by embryogenesis and / or proliferation as part of hydrolysis products and / or cell differentiation, and / or pathology of previous and / or concomitant tissue regeneration and / or repair.

 It should be borne in mind that the term morphoplasma is used here in a broad sense and covers the structured part of the cytoplasm of cells, represented by cellular organs, membranes, the cytoskeleton, and similar cellular structures (Yasus Kagawa. Biomembranes. M. Higher School 1985, p. 13.15 ), and the term glycocalyx is used to refer to a hydrocarbon-containing layer adjacent to the outer side of the plasma membrane of a cell (Alberts B. Bray D. Lewis J. Molecular cell biology. M. Mir, 1986, p. 97).

The problem is solved by the fact that in the modified components morfoplazmy and glycocalyx of cells proposed BAS comprises thermostable at a temperature T _n ≅ ¹²⁰ ° C components. Moreover, the relative content of thermostable components among the modified components is 30-100% by weight.

 In addition, as part of the modified components of morphoplasm and glycocalyx of cells, the proposed ALS contains low molecular weight components with a molecular mass of M М 1.0 kDa. Moreover, the relative content of low molecular weight components among the modified components of the morphoplasm and glycocalyx of the cells is 30-100 wt.

The problem is also solved by the fact that the product contains in the composition of hydrolysis products with modified components of morphoplasm and glycocalyx of cells organic compounds in the following ratio, wt. Polypeptides Up to 26.0 Peptides 10.0-22.5 Amino acids 32.0-60.1 Carbohydrates 10.5-28.7
Lipids, nucleotides
and other organic compounds 2.0-11.0
In this case, the product contains carbohydrates in the composition of organic compounds in the following ratio, wt. consisting of:
High molecular weight
compounds having
_uv molecular mass M> 10.0 kDa up to 2.0
Low molecular weight
unions having
the molecular mass M _un 0.8-10.0 kDa 2.5-11.5
Free monomers M _mind <0.8 kDa 6.0-17.0
In the proposed BAS, the ratio of the mass contents of carbohydrates and peptides in the composition of organic compounds with a molecular mass of M _un 0.8-10.0 kDa lies in the range of G 0.25-0.60.

The problem is solved by the fact that the claimed tool contains in the composition of hydrolysis products with modified components of morphoplasm and glycocalyx of cells organic compounds in the following ratio, wt. Polypeptides 16.3-24.7 Peptides 11.9-17.1 Amino acids 32.0-48.2 Carbohydrates 14.6-20.8
Lipids, nucleotides
and other organic compounds 5.1-9.3.

In addition, the claimed tool contains in the composition of hydrolysis products with modified components of morphoplasm and glycocalyx of cells organic compounds in the following ratio, wt. Polypeptides 5.5-12.7 Peptides 13.6-21.0 Amino acids 37.1-52.9 Carbohydrates 16.1-25.1
Lipids, nucleotides
and other organic compounds 5.3-11.0
The inventive tool also contains in the composition of hydrolysis products with modified components of morphoplasm and glycocalyx of cells organic compounds in the following ratio, wt. Polypeptides Less than 0.3 Peptides 14.9-22.5 Amino acids 43.5-60.1 Carbohydrates 15.5-28.7
Lipids, nucleotides
and other organic compounds 5.0-9.2
The problem is also solved by the fact that in the method of producing BAS, having an immunomodulating property, in which the washed raw material is crushed from animal tissue, the obtained ingredients are hydrolyzed, followed by separation of the hydrolysis products, their filtration and isolation of the supernatant as the target product, they are used as raw materials animal tissue modified during embryogenesis and / or proliferation and / or cell differentiation processes and / or pathology preceding and / or accompanying the regenerator tissue and / or tissue repair, and before hydrolysis, it is homogenized and purified from undamaged tissue elements with the release of a supernatant containing ingredients with components of morphoplasm and glycocalyx cells.

 As raw materials, tissue is used that is selected from the range of: embryonic tissue, tissue of differentiating thymus thymocytes, spermatogonin and spermatocytes of testes, placental tissue, intestinal epithelial tissue, bone marrow cells, regenerating skin tissue, tissue of regenerating liver, tissue of regenerating limbs of amphibians, pathologically altered tissue taken to the stage of regeneration.

 In this case, the cleaning of non-destroyed elements of the tissue after homogenization in physiological and / or buffer solution is carried out by filtration and / or centrifugation at an acceleration of 150-250 g for 10-15 minutes.

 In addition, the grinding of raw materials from animal tissue is carried out before homogenization and lead to damage to the integrity of the cells, followed by extraction of their contents by additional washing with filtration and / or centrifugation at an acceleration of 300-15000 g for 10-20 minutes with the extraction of extracted crushed tissue.

 The problem is solved in that in the composition of the ingredients subjected to hydrolysis, the content of the components of morphoplasm and glycocalyx of cells is 30-60% of the total mass of organic compounds.

Hydrolysis is carried out at a temperature of T _p 4-45 ^about C. In this case, hydrolysis can be carried out in the presence of a preservative.

This object is achieved by the fact that the ingredients of hydrolysis products are subjected to heat treatment at a temperature T _n of 60-120 ^o C to complete denaturation of thermolabile compounds and filtered and / or centrifuged at acceleration of over 1500 g for 20-40 min, the supernatant release comprising thermostable components as the target product.

 In addition, the hydrolysis products of the ingredients are subjected to fractionation by dialysis through a semipermeable film, and / or ultrafiltration, and / or gel filtration to isolate a fraction containing low molecular weight components of morphoplasm and glycocalyx of cells with a molecular mass of M ≅ 10.0 kDa as the target product.

 The problem is solved by the fact that the well-known drug for normalizing the physiological state of the human and animal body, containing a biologically active agent based on the products of hydrolysis of animal tissue ingredients, and a filler, as a biologically active agent, contains thermostable, low molecular weight components of morphoplasm and glycocalyx in the composition of hydrolysis cells modified during embryogenesis and / or proliferation and / or cell differentiation and / or pathology processes, precedence your and / or concomitant regeneration and / or repair of tissue, with its content in the preparation of 0.001-85.0 wt. the rest is filler.

The problem is also solved by the fact that in the preparation a biologically active agent contains organic compounds in the composition of hydrolysis products with modified components of morphoplasm and glycocalyx of cells in the following ratio, wt. Polypeptides Up to 26.0 Peptides 10.0-22.5 Amino acids 32.0-60.1 Carbohydrates 10.5-28.7
Lipids, nucleotides
and other organic compounds 2.0-11.0
In addition, the task can be solved by the fact that BAS contains organic compounds in the ratio, wt. Polypeptides 16.3-24.7 Peptides 11.9-17.1 Amino acids 32.0-48.2 Carbohydrates 14.6-20.8
Lipids, nucleotides
and other organic compounds 5.1-9.3
The problem set in the invention can also be solved by the fact that BAS contains organic compounds in the ratio, wt. Polypeptides 5.5-12.7 Peptides 13.6-21.0 Amino acids 37.1-52.9 Carbohydrates 16.1-25.1
Lipids, nucleotides
and other organic compounds 5.3-11.0
In the proposed drug can also be used biologically active agent containing in its composition, wt. Polypeptides Less than 0.3 Peptides 14.9-22.5 Amino acids 43.5-60.1 Carbohydrates 15.5-28.7
Lipids, nucleotides
and other organic compounds 5.0-9.2
The preparation as a filler may contain a physiological solution of sodium chloride, and / or saline buffer solution, and / or carbohydrate compounds, and / or fats of mineral, animal, vegetable or synthetic origin and mixtures thereof.

 The drug may additionally contain a preservative.

 The problem is solved by the fact that in the known method of normalizing the physiological state of humans and animals in diseases caused mainly by pathologically altered and / or foreign cells, microorganisms and / or products of their vital activity, which involves introducing into the body a preparation based on the products of hydrolysis of animal tissue ingredients, as a preparation, a composition of 0.001-85.0 wt. a biologically active agent containing, as part of hydrolysis products, components of morphoplasm and glycocalyx of cells modified during embryogenesis and / or proliferation and / or differentiation of cells and / or pathology preceding and / or accompanying tissue regeneration and / or repair, and 15 0-99.999 wt. filler, and the introduction of the drug into the body is carried out by intramuscular injection at a dose of 0.005-0.5 mg of the drug per kg of body weight with an interval between injections of 1-7 days and / or inhalation at a dose of 0.012-0.12 mg of the drug per kg of body weight with an interval between inhalations of 4-48 hours, and / or sublingual administration at a dose of 0.012-0.12 mg of agent per kg of body weight with an interval between administrations of 4-48 hours, and / or drinking at a dose of 0.02-1.0 mg funds per kg of body weight with an interval between drinking 1-10 days and / or integrated administration in pills at a dose of 0.02-1.0 mg per kg body weight with an interval between administrations of 1-10 days, and / or intranasal administration in a daily dose of 0.001-0.05 mg of drug per kg of body weight with an interval between administrations of 2-24 hours, and / or application to the mucous or wound surface in the form compresses, suppositories, powders, ointments and gels with intervals between applications of 1-10 days until normalization of physiological state parameters.

 In this case, the normalization of the physiological state of humans and animals is carried out by stimulating the immune system, mainly by activating the reticuloendothelial system. T-, B-systems, neutrophils and / or natural killer populations.

 It has been established that the practical effectiveness of ALS obtained by the proposed method lies in its high biological activity, and above all, in an increased immunomodulating effect (property).

 To a large extent, this determines the strategy for influencing the proposed ALS on the body during normalization of homeostasis parameters.

It is directed by activating the immune system, and above all, cells of the macrophage-granulocyte series, ALS and a drug based on it provide:
1) the impact is not on a specific pathology, but on improving the general status of the physiological state of the body, mobilizing internal reserves aimed at normalizing all parameters of homeostasis, including this pathology. At the same time, concomitant or pathological deviations from the norm without side effects are also corrected. Normalizing the changed parameters, the proposed ALS, unlike many other drugs, does not worsen the remaining parameters of the body's homeostasis;
2) a complex effect that allows you to expand the scope of ALS for a number of diseases, the control of which lies within the competence of the body's immune system;
3) a degree of impact effectiveness aimed at normalizing the parameters of homeostasis, which correlates with the degree of deviation from the norm. In the absence of deviations or normalization of parameters during treatment, the activity of the cells of the immune system stimulated by the drug normalizes within a few days after the administration of ALS without any consequences for the body and, thus, the effect of ALS is terminated. In this case, the feedback principle is realized: the smaller the deviation, the smaller the effect of the effect, and vice versa.

 These results are achieved due to the special properties of ALS and a drug based on it, which are absent in analogues, the manifestation of which is ensured by the presence in the ALS of hydrolysis products containing morphoplasma and glycocalyx components of cells, modified during embryogenesis, and / or proliferation and / or differentiation, and / or pathology preceding and / or concomitant tissue regeneration and / or repair.

 It was found that the increased content of modified components of morphoplasm and glycocalyx of cells is provided by the use of animal tissues taken at the stage of the above mentioned processes of embryogenesis and / or proliferation and / or differentiation and / or pathological processes that preceded and / or accompanied regeneration and / or tissue repair.

These fabrics are:
differentiating thymus thymus cells;
spermatogonia and spermatocytes of the testes, bone marrow cells, intestinal epithelial tissue;
fetal tissue, placental tissue;
tissue of the regenerating limbs of amphibians, regenerating the liver after partial hepatectomy or after being infected with toxic agents, for example, carbon tetrachloride (CCl ₄ ), regenerating skin tissue, as well as tissue with pathology in it, for example, with gangrene, chemical or thermal damage.

 The use of the described animal tissues as raw materials in the proposed method provides BAS with the expected properties.

 It was also established that a characteristic feature of the above types of tissues is the presence in them of modified components of morphoplasm and glycocalyx of cells, which makes it possible to differentiate them from intact or resting cells of adult tissue. This feature of the modified components of the morphoplasm and glycocalyx of the cells of these tissues allows you to use them in the proposed ALS and a preparation based on it, with the introduction of which into the body of an animal or person it is possible to stimulate a directed response of the body to pathology. A key role in this process is played by the body's immune system.

 Material carriers of information in the above processes and / or abnormalities in these tissues are specific components that are part of the structure of the morphoplasm and glycocalyx of cells, which undergoes a modification when the physiological status of cells changes. The proposed ALS and a drug based on it contain modified components of the morphoplasm and glycocalyx of the cells, which allows using them to stimulate the above response of the body.

 Performing before the hydrolysis of the process of washing the crushed tissue from the components of the tissue and blood extract in combination with homogenization, after which they are cleansed of the undamaged tissue elements, provides in the proposed ALS and the preparation based on it a high concentration of the components of the morphoplasm and glycocalyx of cells (30-60%), as well as a significant decrease in the content of immunosuppressants, neutral (in relation to specific activity) and ballast compounds that make up the remainder of the mass in the target product. This enhances the directed immune response of the body.

Response of the organism is enhanced significantly when administered the proposed agent which contains in its composition components morfoplazmy and glycocalyx of cells in the form of thermostable components (stable at the heating temperature T _n ≅ ¹²⁰ ° C) as a low-molecular components with a molecular mass M ≅ 10.0 kDa .

 The proposed ALS is used as an active principle in a preparation in which various excipients are used depending on the purpose.

 For internal use, the drug is prescribed in the form of injections and the excipient is a physiological solution of sodium chloride or buffered saline. For external use, in addition to the listed solutions, gel components include carbohydrate compounds of animal and plant origin, for example, honey, juices, plant flesh and the like, or ointments of mineral, animal, plant or synthetic fats. and mixtures thereof.

 In this case, an unobvious dependence between the effectiveness of the drug and the content of ALS was found. This made it possible to significantly reduce the content of the active ingredient in the drug while maintaining the necessary effectiveness of the drug and thereby prevent the possibility of overdose, the occurrence of side effects, as well as significantly reduce its cost.

 In addition, it was found that the new properties of the drug described above make it possible to use it to normalize the physiological state of the body when it is introduced into the body in almost any dose, namely from 0.001 to several grams per 1 kg of body weight.

 These properties are illustrated below in the examples of specific implementations of the invention, in particular in examples 1,2,4,8,9,12,16, 18-25 and others.

 In the above method, due to the combined factors of using the new drug, its specific dosages, the frequency of its introduction into the body, as well as the methods of introducing it into the human and animal body, the treatment of diseases caused mainly by pathologically altered and / or foreign cells, microorganisms and / or their products is provided. life, as illustrated below in the examples of specific use of the invention, in particular in examples 3,4,7,8,11-13, 15-17 and others.

 According to the invention, a biologically active agent having an immunomodulatory property is obtained as follows for normalizing the physiological state of an organism.

 The raw material is animal tissue modified during embryogenesis and / or proliferation and / or differentiation and / or pathology preceding and / or concomitant tissue regeneration and / or repair. The specified animal tissue is washed and crushed, after which it is homogenized in physiological and / or buffer solution until the cells are completely destroyed, and the homogenized mixture is cleaned of intact tissue elements by filtration and / or centrifugation with the release of ingredients containing morphoplasma and glycocalyx components of cells, and subjected to hydrolysis. The products of hydrolysis after centrifugation and / or filtration are used as the target product to normalize the physiological state of the body, in particular for external use (inhalation, application, intranasally, as part of an ointment, and so on).

 The effectiveness of the target product depends on its content of modified components of morphoplasm and glycocalyx of cells. Increases in their content are achieved by the selection of raw materials with increased intensity of the processes of modification in weaving and the selection of optimal conditions for the implementation of the processing method.

 The choice of raw materials is determined by the fact that the components of the morphoplasm and glycocalyx of the cells react quite specifically to changes in the physiological state of cells by changing their structural organization, which lead to a change in the immunogenicity of the tissue, and thereby cause corresponding reactions of the body's immune system. It was established that the immunogenicity of the modified structure is provided at the stage of the above processes (embryogenesis and the like).

As animal tissue use, in particular:
differentiating thymus thymocytes (cell differentiation process);
spermatogonia and spermatocytes of the testes, bone marrow cells, intestinal epithelial tissue (proliferation and differentiation of cells);
embryonic tissue, placental tissue (processes of polyperation, differentiation, embryogenesis);
tissue of the regenerating limbs of amphibians, the regenerating liver of rats after partial hepatectomy or after being struck by carbon tetrachloride (CCl ₄ ), regenerating skin tissue, as well as tissue with pathology in it, for example, with gangrene, chemical or thermal damage (a combination of various combinations of the above processes) .

 The use of the described animal tissues as raw materials in the manufacturing method provides a biologically active agent with the expected properties.

 It has been established that a characteristic feature of the above types of tissues is a change in the structure of the components of morphoplasm and glycocalyx of tissue cells in them, which makes it possible to differentiate them from intact or resting cells of adult tissue. This feature of the modified components of the morphoplasm and glycocalyx of the cells of these tissues makes it possible to use them in ALS, when introduced into the body of an animal or human, it is possible to stimulate a directed response of the organism to pathology. A key role in this process is played by the body's immune system.

 It is preferable to use animal tissue taken at the stage of increment of the modification process. This stage is determined empirically by the nature of the change in immunogenicity. So, in the case of using embryonic tissue, this period is the first half of embryogenesis. The regenerating liver of rats is used at 4-12 hours after partial hepatectomy, and in the case of using regenerative limbs of amphibians at the stage of blastema formation.

 An increase in the relative content of the components of the morphoplasm and glycocalyx of the cells is achieved due to the implementation of such a regime of the claimed method, one of the indispensable conditions for the implementation of which is the removal of intact tissue elements, represented mainly by connective tissue.

 Purification from undisturbed tissue elements is provided during centrifugation in the acceleration range of 150-250 g for 10-15 minutes, which is explained by the fact that at higher accelerations and longer process duration, cells of morphoplasm and glycocalyx are precipitated, and at accelerations less than 150 g cleaning is ineffective.

 Purification from non-destroyed elements is also provided by filtration on a large-pore filter, for example through several layers of gauze, through a cotton cloth (calico) or through nylon mesh.

 To enhance the immunomodulatory properties of the target product before homogenization, the tissue is crushed to damage the integrity of the cells, followed by extraction of their contents with additional washing with physiological saline and / or organic acid solution by filtration and / or centrifugation under acceleration of 300-15000 g for 10-20 min and extracted crushed tissue, which is subjected to homogenization.

 In this case, on the one hand, they achieve the removal of a significant part of the immunosuppressants present in the blood serum and tissue extract, and on the other hand, increase the relative content of the components of morphoplasm and glycocalyx of cells, which is essential for tissues with a low content.

 Additional washing of the used ground animal tissue before homogenization by centrifugation at 300-15000 g for 10-20 minutes allows you to most effectively separate the extracted tissue from the extract under industrial conditions. A similar effect is achieved by filtration on a large-pore filter, for example, through several layers of gauze, through cotton fabric (calico) or through nylon mesh.

 The relative content of the components of the morphoplasm and glycocalyx of the cells in the tissue ingredients isolated for hydrolysis is determined by the content of proteins and carbohydrates in the cell structures freed from the intact tissue elements.

 The effectiveness of the target product significantly increases when using components of morphoplasm and glycocalyx of cells in the form of low molecular weight components. This allows, on the one hand, to avoid the manifestation of undesirable side effects, and on the other hand, to obtain a broad-action drug without limiting the species-specificity of animals and meeting the requirements of medicine. To this end, tissue ingredients containing modified components of morphoplasm and glycocalyx of cells were subjected to hydrolysis, splitting high molecular weight structural formations of morphoplasm and glycocalyx of cells into products containing components with a lower molecular weight. The effectiveness of the hydrolysis products will depend on the specificity of the hydrolysis, which largely depends on the temperature-time characteristics.

It is found that the immunomodulatory properties of the proposed agent, in particular an increase of activity in vitro of macrophages of rat peritoneal exudate maximum in the case of hydrolysis in the operating temperature range of T ₌ 4-45 ^° C

Deviations from it are undesirable, since:
at T _p <4 ^{° C} significantly increases the duration of the hydrolysis process that reduces the economic efficiency;
_p at T> ⁴⁵ ° C, the reduction in the activity of enzymes ensuring hydrolysis of raw material. and accelerate the decay of active components. As a result, its quality is reduced.

 The duration of hydrolysis is determined by reaching the maximum level of immunostimulating effect (properties) of the product, in particular, by activating macrophages of peritoneal exudate according to the HCT test.

 Prolonged hydrolysis is carried out in the presence of a preservative.

 The increase in the relative content (relative concentration) of low molecular weight components in the hydrolysis products with respect to the content of high molecular weight thermolabile compounds is provided by heat treatment of the hydrolyzate until the thermolabile compounds are completely denatured, which are then removed as a precipitate by centrifugation with acceleration of at least 1500 g for 20-40 min and / or by filtration. The supernatant containing thermostable components of morphoplasm and glycocalyx cells is used as the target product.

The thermal treatment of the hydrolysis products is carried out in the temperature range T _n = 60-120 ^{° C,} since the yield of this range leads to a decrease in specific activity means.

Thus, at a temperature T _n <60 ^{° C,} not all proteins undergo denaturation, resulting in the final product the content of thermolabile compounds will be increased. Due to this, the specific activity of the agent will be reduced. When the temperature T _H> ¹²⁰ ° C causes an abrupt increase in rate of structural changes in peptide and carbohydrate components of desired product, which also reduces its quality.

 After heat treatment, the selection of thermostable components is carried out by centrifugation at an acceleration of at least 1500 g for 20-40 minutes, since with a decrease in this value, the degree of removal of denatured thermolabile compounds is significantly reduced, and holding it for 20-40 minutes is the most optimal.

 Coagulated denatured thermolabile compounds can be separated from the supernatant by filtration through a paper filter.

 The target product containing the thermolabile components of morphoplasm and glycocalyx of the cells has greater specific activity and allows to solve a wider range of tasks. It can be used as an injection drug in both veterinary medicine and in medicine.

 A further increase in the specific activity of ALS is achieved by isolating low molecular weight components with a molecular mass of M ≅ 10.0 kDa from hydrolysis products by dialysis through a semi-impermeable film, and / or ultrafiltration, and / or gel filtration, which completely removes high molecular weight compounds that can cause side effects.

 The value of efficiency achieved in this way makes it possible to widely use the obtained target product in medicine.

 The proposed ALS as an active principle is used for the preparation of a medicinal product, where its concentration depends on the purpose of use. In preparations for feeding and feeding animals, the concentration of ALS is low and increases in preparations for injection use, as well as in ointments, gels and powders.

 Depending on the purpose, various excipients are used in the preparations. So, in preparations for injection, inhalation, applications and intranasal use, a physiological solution of sodium chloride or saline buffer solution is used. Carbohydrate compounds are used as filler in preparations intended for feeding bees (for example, 10% sugar syrup) or for preparing gels (for example, honey, juices and pulp of plants). For ointments, fats of mineral, animal, vegetable or synthetic origin and mixtures thereof are used as filler.

PRI me R 1. To obtain ALS was used embryonic tissue taken at the stage of the first half of pregnancy of cows. Muscle and skin tissue, liver, spleen, pancreas, kidneys, lungs, and heart were cut from embryos after disinfection in a weak solution of potassium permanganate (KMn ₇ O ₄ ), which were subjected to rough grinding in a meat grinder.

Then 10 kg of minced meat was mixed with 30 kg of saline and homogenized on a homogenizer with knife blenders until a homogeneous mass was obtained. Purification of the mixture of non-disintegrated tissue elements carried by decantation, and the resulting supernatant with a tissue ingredients containing components morfoplazmy and glycocalyx of cells, when subjected to autolysis _TH 45 ^{° C} for 48 hours.

 The obtained hydrolyzate was filtered on a paper filter, and the filtrate containing the components of morphoplasm and glycocalyx of the cells was used as the target product, the composition of the organic compounds of which and the specific activity caused by it are presented in Tables 1 and 2.

 The target product containing ALS in the amount of 1.0 wt. and the rest is a filler, mixed with 9 volumes of cucumber pear pulp and used as a medicine in the form of a gel mask for the face for cosmetic purposes. During prolonged storage, a preservative (nipagin and nipazole in a mass ratio of 5: 1) was added to the gel at a final concentration of 0.1 wt. The gel was applied to the face for 2-3 hours. The procedure was repeated 1-2 times a week. The therapeutic effect was manifested after 4-10 sessions and it was controlled by the condition of the skin.

PRI me R 2. Embryonic tissue obtained in the same manner as in example 1, was subjected to grinding. Then 10 kg of minced meat was mixed with 30 kg of saline and homogenized until a homogeneous mass was obtained. To remove undamaged tissue elements, including connective tissue elements, the homogenized mass was centrifuged at an acceleration of 250 g for 10 min. After centrifugation, the precipitate was removed, and the supernatant was filtered through cotton (calico) to remove flotated fat. In the filtrate of the thus purified homogenate containing 30 wt. components of morphoplasm and glycocalyx cells, a preservative (chinosol) was added to a final concentration of 0.08 wt. After mixing, the homogenate was subjected to hydrolysis at a temperature of T _p 4 ^about C for 6 weeks.

 The obtained hydrolyzate was centrifuged at an acceleration of 15,000 g for 20 min, and the supernatant with the components of the morphoplasm and glycocalyx of the cells was isolated as the target product, the composition of the organic compounds of which and the specific activity caused by them are presented in Tables 1 and 2.

The target product containing ALS in the amount of 1.0 wt. and the rest was used as a medicine to increase the safety of weakened young fur-bearing animals, as well as poultry chickens in the form of inhalation in a special chamber where the animals were kept for 15-20 minutes. In the chamber, the drug was sprayed aerosol at the rate of 0.5 ml per 1 m ³ for 1 min. The procedure was repeated every 7 days. The control of the effect was carried out by reducing the death rate of young animals, which decreased by 30-40%
PRI me R 3. The embryonic tissue obtained in the same manner as in example 1 was subjected to grinding. The resulting stuffing was poured with a double volume of saline and mixed. The mixture was centrifuged at an acceleration of 1500 g for 20 minutes. The precipitate was recovered, mixed again in a double volume of physiological saline and centrifuged under the same conditions. Forcemeat washing operation was carried out 3 times. Then, 10 kg of washed minced meat was mixed with 30 kg of a centimolar phosphate saline buffer solution (0.01 M PBS) pH 7.2 containing a centimolar disubstituted sodium phosphate and a fifteen centimolar sodium chloride solution (0.01 M Na ₂ HPO ₄ and 0.15 M NaCl), and subjected to homogenization in a colloidal mill to obtain a homogeneous mass. Undestructed tissue elements were removed as a precipitate after centrifugation at an acceleration of 150 g for 15 minutes. The supernatant after removal of the flotated fat by filtration through a cotton cloth (calico) was treated with a preservative with a final concentration of 0.08 wt. After mixing, a homogenate containing 60 wt. components of morphoplasm and glycocalyx cells, were subjected to hydrolysis at a temperature of T _p 4 ^about C for 6 weeks.

 The obtained hydrolyzate was centrifuged at an acceleration of 1500 g for 40 min with the isolation of a supernatant containing the components of morphoplasm and glycocalyx of cells as the target product, the composition of the organic compounds of which and the specific activity caused by them are presented in Tables 1 and 2.

 The target product containing ALS in the amount of 1.0 wt. and the rest of the filler was used as a medicine for the treatment of calves suffering from parainfluenza by injection, which was administered intramuscularly at a dose of 0.05 ml / kg of body every third day. The therapeutic effect was manifested after 1-5 injections and it was controlled by body temperature and the presence of the virus in the blood.

PRI me R 4. Coarsely ground in a meat grinder, obtained in the same way as in example 1, was poured with a double volume of 3% acetic acid and mixed. The mixture was filtered through cotton under vacuum on a suction filter. The precipitate was removed, again mixed in a double volume of physiological saline and centrifuged at 300 g for 20 minutes. 10 kg of washed minced meat was mixed with 30 kg of a centimolar phosphate saline buffer solution (0.01 M PBS) pH 7.2 containing a centimolar bisubstituted sodium phosphate and a fifteen centimolar sodium chloride solution (0.01 M Na ₂ HPO ₄ and 0.15 M NaCl ) and subjected to homogenization in a colloidal mill to obtain a homogeneous mass. To remove undamaged tissue elements, the homogenized mass was filtered through cotton fabric (calico). In the obtained filtrate was added a preservative in a final concentration of 0.08 wt. After mixing, a homogenate containing 50 wt. components of morphoplasm and glycocalyx cells, were subjected to hydrolysis at a temperature of T _p 20 ^about C for 8 days. Further operations were carried out as in example 3.

 The content of organic compounds in the target product and indicators of its specific activity, determined experimentally, are given in tables 1 and 2.

 The target product containing ALS in the amount of 1.0 wt. and the rest of the filler was used as a medicine for the treatment of dermatoses in the form of an ointment, which was prepared by mixing lanolin, petroleum jelly and the target ALS product in a ratio of 1: 3: 1 mass parts. The ointment was rubbed into the skin daily. The therapeutic effect was manifested after 4-12 days and it was controlled by the condition of the skin.

 PRI me R 5. The relative content of the components of morphoplasm and glycocalyx of cells as a percentage (by weight) to the components of connective tissue and extract in the tissue ingredients isolated for hydrolysis, was determined by the content of proteins and carbohydrates in cellular structures freed from intact fabric elements.

 For this purpose, the homogenate, purified as in Examples 1-4, from undestructed tissue elements, was subjected to ultracentrifugation at an acceleration of 100,000 g for 60 minutes.

 The ratio of the amount of protein in the whole sediment and in the whole supernatant indicates the ratio of the components of the morphoplasm and glycocalyx of the cells to the proteins of the extract.

 Determination of protein content in sediment and supernatant was carried out after ultracentrifugation of the homogenate. After removal of the supernatant, 4.9 ml of physiological saline was added to 0.1 ml of the precipitate, and after suspension 0.1 ml was taken to determine the amount of protein photometrically from the color of the solution according to the SDS-Lowry method (UKLaemmli. Nature, 1970, v.227, 5259, p. 680-685).

 To determine the amount of protein in the supernatant, 0.9 ml of physiological saline was added to 0.1 ml of the supernatant, and after stirring, 0.1 ml was taken to determine the amount of protein by SDS Lowry.

 The content of connective tissue components was determined by amino acid analysis by the presence of oxylysine and oxyproline. Unlike the known ones, the proposed ALS did not contain amino acid data during the test.

 The content of glycocalyx components of cells was determined by the amount of carbohydrate in the homogenate, which are part of compounds with a molecular weight of more than 0.8 kDa, according to the anthrone reaction after removal of glycogen granules from the homogenate (S. Seifter, S. Dayton, C. Hovic Arch. Biochemistry and Biophysical , 1950, 25, 1. R. 191-200).

 For this purpose, 20 ml of tissue homogenate was layered in ultracentrifuge tubes with a capacity of 30 ml per 5 ml of sucrose with a density of d 1.40 and subjected to ultracentrifugation at an acceleration of 100,000 g for 60 minutes. The glycogen deposited on the bottom of the tube was removed, and the elements of morphoplasm and glycocalyx of the cells were taken at the interphase and after washing from sucrose by centrifugation in physiological saline was used for analysis.

 When determining the amount of carbohydrates in the studied samples, the total carbohydrate content after hydrolysis and the content of free carbohydrates in the form of monomers were taken into account. The difference in the values of this value in the measurements determined the amount of free carbohydrates in the composition of various compounds.

To carry out general hydrolysis, 1.0 ml of the test sample was taken and mixed with 1 ml of a tetra-normal hydrochloric acid solution (4.0 NHCl). Hydrolysis was performed at 105 ^{° C} for 2 hours after which the hydrolyzate was evaporated in the exsiccator with dry sodium hydroxide (NaOH) under vacuum. The dried sample was dissolved in 1.0 ml of distilled water and used for analysis.

Determination of the number of free carbohydrates in a BAS directly or after acid hydrolysis was performed as follows: 1 ml samples were mixed with 1 ml of distilled water, then under constant stirring at -5 ^{° C} was added 4 ml of 0.2% anthrone reagent prepared in 95% sulfuric acid (H ₂ SO ₄ ). The resulting mixture was boiled at a temperature of 95 ^about C for 10 min and after cooling, the optical density was determined by a wavelength of 620 nm. Glucose at a concentration of 20 μg / ml was used as a standard.

In the protein analysis of ALS obtained in examples 1 and 2, the content of the components of the morphoplasm and glycocalyx of the cells was 37.6 7.4% and for ALS in examples 3 and 4 51.8 ± 8.2%
Hereinafter in the text and in the tables below, the value of experimentally determined values is expressed by their average values with the deviation limits from the average, calculated taking into account the Student criterion.

EXAMPLE Example 6 The hydrolyzate supernatant, obtained in Example 3 was subjected to heat treatment at heating temperatures (heat treatment): T _N1 = 55 ^{° C,} T 60 ^o C _H2, T _H3 95 ^o C, T _H4 ¹²⁰ ° C and T _n5 135 ^about C.

Heating was carried out until the thermolabile compounds were completely denatured as part of the hydrolysis products, then they were centrifuged at an acceleration of more than 1500 g for 30 min with the release of a supernatant containing thermostable components of morphoplasm and cell glycocalyx as the target product. In each case, the obtained target product had its own immunomodulating effect (property), the values of which are shown in Table 3, which shows the distribution of the values of the specific activity of the proposed BAS depending on the selected range of heating temperature of the hydrolyzate and outside this range. When this is the optimum heating temperature T _H ^of 95 C., at which produced the desired product provide maximum gain and figotsitarnoy enzymatic activity, which was respectively 72.8% and 64.3% Three T _n 55 ^C values increase in specific activity compared to control is an average of 61.1% for the HCT test and 52.3% for phagocytosis. At T _n > 120 ^° C, an abrupt increase in the rate of structural changes in peptides and hydrocarbon-containing compounds occurs, leading to structural changes in the components of the target product, which is reflected in a decrease in the specific activity of ALS. Thus, at T _n 135 ° ^C an increase in specific activity of the present BAA compared to the control is an average of 57.3% by NBT-test and 43.0% by phagocytosis. The maximum value of BAS specific activity compared to the control is provided in the temperature range ^of 60-120 _N T C at which the increased activity of macrophages peritone- cial exudate of rats (in vitro) is on average 72.8% to enhance the enzymatic activity (HCT-test ), and by enhancing phagocytic activity (by E. Coli) 64.3%
Example 7. The supernatant of the hydrolyzate obtained in Example 3 was subjected to heat treatment by boiling at a heating temperature of Т _н 95 ^о С until the thermostable compounds were completely denatured and, after cooling, they were centrifuged at an acceleration of 1500 g for 40 minutes. The precipitate in the form of denatured compounds was removed, and the supernatant was used as the target product, which contained thermostable components in an amount of 60 wt. according to peptides in relation to the products of hydrolysis of water-soluble compounds before heat treatment.

 The content of organic compounds in the hydrolysis products and the specific activity indicators of the obtained agent, determined experimentally, are shown in Tables 1 and 2.

 The target product containing ALS in the amount of 0.5 wt. and the rest of the filler was used as a drug for the treatment of mastitis in cows in the form of injections, which were carried out intramuscularly at a dose of 0.05 ml / kg body weight every third day. The therapeutic effect was manifested after 1-4 injections and was contacted by the condition of the udder and the number of purulent discharge from the nipples.

EXAMPLE Example 8 The hydrolyzate supernatant, obtained in Example 2 was subjected to a heat treatment at ^about T _p 95 C until complete denaturation of thermolabile compounds and, after cooling, centrifuged at 50,000 g for 20 min. The precipitate was removed, and the supernatant was used as the target product, which contained thermostable components in an amount of 30 wt. according to peptides in relation to the products of hydrolysis of water-soluble compounds before heat treatment. The content of organic compounds in the hydrolysis products and the specific activity indicators of the obtained agent, determined experimentally, are shown in Tables 1 and 2.

The resulting target product containing ALS in the amount of 0.5 wt. and the rest of the excipient was used as a medicine for the prevention of calves with parainfluenza in the form of injections, which were administered intramuscularly at a dose of 0.05 ml / kg body weight twice with an interval of 14 days. The effectiveness of the drug was recorded by the number of sick animals within a month, which in the experimental group did not exceed 10% whereas in the control it reached 22-28%
PRI me R 9. The hydrolyzate obtained in example 4 was subjected to heat treatment at a temperature of T _p 95 ^about With until complete denaturation of thermolabile compounds and after cooling was filtered through filter paper on a suction filter. The filtrate containing thermostable components in an amount of 45% of the peptides with respect to the hydrolysis products before the heat treatment of water-soluble compounds was used as the target product.

 The content of organic compounds in the hydrolysis products and the specific activity indicators of the obtained agent, determined experimentally, are shown in Tables 1 and 2.

 The target product was freeze-dried and used sublingually as a drug for the treatment of mastitis in women. The procedure was performed daily. The therapeutic effect was manifested after 1-4 days and was monitored by the state of the chest (by the presence of seals, purulent discharge) and a blood test.

 Example 10. The supernatant of the hydrolyzate obtained in example 3 was subjected to fractionation on an Amicon ultrafiltration unit using ultrafilters transmitting compounds with a molecular weight of less than or equal to: 50.0 kDa; 10.0 kDa; 5.0 kDa; 1.0 kDa; 0.8 kDa and 0.5 kDa.

The biological activity of the obtained fractions containing compounds with a molecular weight of: 1) M ₁ > 50.0 kDa; 2) 10.0 kDa <M ₂ <50.0 kDa; 3) M ₃ <10.0 kDa; 4) M ₄ <5.0 kDa; 5) M ₅ <1.0 kDa; 6) M ₆ <0.8 kDa and 7) M ₇ <0.5 kDa, were analyzed by their ability to activate macrophages of rat peritoneal exudate according to the HCT test. The research results are summarized in table 4.

Analysis of the data in table 4 allows us to draw the following conclusions:
1) the distribution of specific activity by fractions has a pronounced maximum, which falls on the molecular weight range from 0.8 to 10.0 kDa;
2) a decrease in specific activity at a molecular mass of M> 10.0 kDa is associated with an increase in the influence of inhibitors in the target product and a decrease in the concentration of the active substance;
3) a decrease in specific activity at a molecular mass of M <0.8 kDa is associated with a decrease in the concentration of the active substance, presented in the form of oligomeric compounds with M> 0.8 kDa.

 PRI me R 11. The supernatant of the hydrolyzate obtained in example 3 was subjected to ultrafiltration on hollow fibers VPU-15-PA (TU 6-12-151-87) apparatus AP-2.0. The resulting ultrafiltrate containing low molecular weight components of morphoplasm and glycocalyx of cells with M <10.0 kDa was used as the target product, where low molecular weight components with M ≅ 10.0 kDa were 60 wt. peptides in relation to the products of hydrolysis of water-soluble compounds before ultrafiltration.

 The content of organic compounds in the hydrolysis products and the specific activity indicators of the obtained agent, determined experimentally, are shown in Tables 1 and 2.

 The target product containing ALS in the amount of 0.1 wt. and the rest of the excipient was used as a drug for the treatment of experimental rat hepatitis in the form of injections, which were administered intramuscularly at a dose of 0.05 ml / kg body weight every other day (the nature and results of the treatment are described in more detail in examples 34 and 35).

 Example 12. The supernatant of the hydrolyzate obtained in Example 2 was dialyzed for 16-24 hours through a cellophane film against physiological saline, 1 kg of which contained 0.8 g of preservative. The resulting dialysate was used as the target product, which contained low molecular weight components with M ≅ 10.0 kDa in an amount of 30 wt. peptides in relation to the products of hydrolysis of water-soluble compounds before dialysis. The content of organic compounds in the hydrolysis products and the specific activity indices of the obtained ALS, determined experimentally, are shown in Tables 1 and 2.

 The target product containing ALS in the amount of 0.1 wt. and the rest of the excipient was used as a medicine in the form of injections to calves to normalize the parameters of homeostasis. Injections of the drug to calves with abnormalities were carried out intramuscularly at a dose of 0.05 ml / kg body weight every 1-2 months. The effect was controlled by monthly weighing of animals, the results of which showed a 45–70% increase in average daily gain compared to the control group of untreated animals.

 PRI me R 13. The supernatant of the hydrolyzate obtained in example 7 was subjected to gel filtration in Sephadax G-25 on a column with a diameter of d 26 mm and a length of h 600 mm, balanced with decimolar (0.01 M) phosphate buffer solution pH 7 , 2 containing fifteen centimolar sodium chloride (0.15 M NaCl). The fractionation of the hydrolyzate supernatant by molecular weight was carried out by eluting the column with the same buffer solution and the chromatographic profile of the eluate was controlled spectrometrically at a wavelength of 280 nm. The fraction of low molecular weight components of the hydrolyzate containing the components of morphoplasm and glycocalyx of cells was eluted as the target product. The content of organic compounds in the hydrolysis products and the specific activity indicators of the obtained agent, determined experimentally, are shown in Tables 1 and 2.

 The target product containing ALS in the amount of 0.1 wt. and the rest of the excipient was used as a medicine for the treatment of calves suffering from bronchopneumonia or acute respiratory diseases (ARI), in the form of injections, which were administered intramuscularly at a dose of 0.05 ml / kg body weight every third day. The therapeutic effect, which was manifested after 1-7 injections, was controlled by body temperature.

 PRI me R 14. The dependence of the effectiveness of the claimed ALS on the content of modified components of morphoplasm and glycocalyx of cells in it was determined using raw materials from animal tissues in which embryogenesis and / or proliferation and / or cell differentiation and / or pathology took place preceding and / or concomitant tissue regeneration and / or repair. The resulting tissue was processed in the same way as in example 7. The properties of the target product obtained in this case, that is, using the tissue of each of these types of modifications of cell structures, namely morphoplasm and glycocalyx of cells, are summarized in table 5.

 Analysis of Table 5 shows that the presence of an immunomodulating effect in the proposed ALS and a significant increase in this effect depend on the raw materials used, from which hydrolysis products containing modified components of morphoplasm and glycocalyx of cells were obtained.

So, spermatogonia and spermatocytes of rat testes, bone marrow cells, rat intestinal epithelial tissue were used as raw materials from animal tissue taken precisely at the stage of cell proliferation and differentiation, and rat embryonic tissue was used as raw material with embryogenesis processes. The liver tissue of rats after partial hepatectomy was used as a raw material with pathological processes prior to regeneration, and the liver tissue of rats after subcutaneous injection of carbon tetrachloride (CCl ₄ ) was used as a raw material with pathology associated with regeneration of the tissue of the regenerating limbs of axolotl.

 Simultaneously with the obtained experimental data, Table 5 also compares for comparison the information on the immunomodulating effect obtained by using adult male rats as muscle and liver intact tissue as raw materials.

 An analysis of the data in Table 5 shows that the immunomodulating effect of ALS when using tissues taken at the stage of embryogenesis, proliferation, and pathology that preceded and / or accompanied regeneration and / or repair was significantly higher than that of known agents. using raw materials from animal tissue in which these processes were absent.

 In addition, we can also conclude that of the tissues in which these processes are observed, the regenerating liver tissue, embryonic tissue, and regenerating tissue of the amphibian limbs have the greatest immunomodulating effect.

 PRI me R 15. As a raw material used regenerating rat liver at 4-12 hours after hepatectomy performed according to the known method (GM Higgins, RMA Anderson Experimental pathology of the liver. Arch. Pathology, 1931-N 12-p. 186).

 Liver tissue was finely cut and mixed with a three-fold volume of physiological saline, after which it was centrifuged at an acceleration of 15,000 g for 10 min, and the resulting precipitate was mixed with a three-fold volume of saline and centrifuged under those conditions. The washing operation of the crushed liver tissue was repeated 3 times and then further stages of obtaining the target product were performed in the same way as in examples 3.7 and 11.

The specific activity of the obtained target products was determined experimentally by activating the repair ability of rat liver hepatocytes after being affected by carbon tetrachloride (CCl ₄ ), in which the activity of Na, K-ATPase was restored, respectively, by 32.7 ± 3.1% 43.6 ± 3, 9% and 85.4 ± 7.8% compared with the control group of animals to which the proposed tool was not administered.

The activity of aminotransferases in the blood was restored, respectively, by 37.4 ± 2.8% 47.2 ± 3.4% and 98.3 ± 1.7%, and the concentration of potassium in liver hepatocytes by 22.6 ± 1.8% 28 , 9 ± 2.5% and 59.1 ± 4.7%
The target product containing ALS in the amount of 0.1 wt. and the rest of the filler was used as a drug for the treatment of hepatitis calves in the form of injections, which were administered intramuscularly at a dose of 0.05 ml / kg body weight every 3rd day. The therapeutic effect was manifested after 7-14 injections and it was controlled by the activity of aminotransferases, the concentration of bilirubin and nitrogen in the blood.

 PRI me R 16. As a raw material used the seed testicles of bulls obtained after isolation of lidase from them by extraction of 3% acetic acid tissue homogenate.

 The obtained seed cake, which is a waste of lidase production, was re-homogenized in a triple volume of physiological saline on knife homogenizers and centrifuged at 250 g for 15 minutes to remove undamaged tissue elements. Using a decinormal aqueous solution of caustic soda (0.1 N NaOH), the acidity of the medium was adjusted to pH 7.4 and a preservative of 0.8 g / l was added to the homogenate containing ingredients with morphoplasma and glycocalyx components of cells modified during proliferation and differentiation of cells, which, after mixing, was subjected to hydrolysis and subsequent processing in the same way as in examples 3.7 and 11.

 The specific activity of the obtained target products was determined experimentally by the activation of macrophages of peritoneal exudate and rat blood neutrophils. When using the target product obtained in the same way as in Example 11, the enzymatic activity of macrophages increased by 108.3 ± 13.2% and the phagocytic activity by E. coli by 92.2 ± 10.2%. For blood neutrophils, these indices increased by 99.2 ± 14.1% and 72.4 ± 8.3%, respectively.

 The target product containing ALS in the amount of 1.0 wt. and the rest is a filler, mixed with 50 volumes of water and used as a drug in the form of a drink to normalize the parameters of chicken homeostasis. Drinking chickens with deviations of the indicated parameters from the norm was carried out at a dose of 5 ml / kg body weight every 2-6 weeks. The effect was controlled by calculating the egg production, the results of which showed its increase by 15-30% compared with the control group of birds not treated with the drug.

PRI me R 17. As a raw material used placental tissue of cows, which was processed in the same way as in examples 3, 7 and 11, to obtain the target products, the specific activity of which was determined experimentally by the activation of macrophages of peritoneal exudate and rat blood neutrophils in comparison with the control. When using the target product obtained in the same way as in Example 11, the enzymatic activity of macrophages increased by 150.6 ± 17.4% and the phagocytic activity of macrophages increased by 109.3 ± 12.7%. For blood neutrophils, the corresponding indices were 137.6 ± 16.8% and 95.7 ± 11.3%
The target product containing ALS in the amount of 0.5 wt. and the rest of the filler was used as a drug for the treatment of cows with endometritis in the form of injections, which were administered intramuscularly at a dose of 0.05 ml / kg body weight every 3rd day. The therapeutic effect was manifested after 6-10 injections and was monitored according to the indicators of the inflammatory reaction, histological studies and purulent discharge.

 PRI me R 18. For the manufacture of the proposed BAS as a raw material used thymus cake obtained after isolation from the homonenate of insoluble tissue components, which are a waste product of thymosin.

 Thymus cake was treated in the same manner as in examples 3.7 and 11 to obtain the target products, the specific activity of which was determined experimentally by stimulating the T-lymphocyte blast transformation reaction, which increased by 165.6 ± 13.7% compared to the control at using the target product obtained in the same manner as in example 11.

 The target product containing ALS in the amount of 0.5 wt. and the rest of the excipient was used intranasally as a drug for the prevention of human infectious diseases. 2 drops of the drug were introduced into each nostril followed by massage for spreading along the mucous membrane. This procedure was repeated 2 times a day. The effect was controlled by the general condition of the body and body temperature.

PRI me R 19. A drug based on ALS obtained in the same way as described in examples 1-4, 7-9, 11-13 and 15-17, was prepared with the content of the specified ALS in the drug in an amount of 0.001 wt. The filler was a 10% sucrose solution. The drug was used to feed the bees. One-time top dressing in the amount of 1 liter of the drug per bee family provided, compared with the control, an increase in survival by 85-140%, an increase in the number of departures by 100-160%, an increase in honey collection by 27-64%
PRI me R 20. A drug based on ALS obtained in the same way as described in examples 1-4, 7-9, 11-13 and 15-17, was made after freeze drying the specified ALS with its content in the drug in the amount of 85.0 wt. The filler was an antiseptic. The drug in the form of powder from lyophilized powder was used to treat open wounds for burns, injuries and erosion of the mucosa. The use of the drug accelerated regenerative processes by 2-7 times in comparison with the control.

 PRI me R 21. A drug based on ALS obtained in the same way as described in examples 1-4, 7-9, 11-13 and 15-17, was made with the content of the specified ALS in the drug in an amount of 1.0 wt. The filler was nel from vegetable pulp such as cucumber, apple, and peach puree. The drug was used for cosmetic purposes. The therapeutic effect was manifested after 4-8 sessions and was controlled by skin elasticity.

 PRI me R 22. A drug based on ALS obtained in the same way as described in examples 1-6, 9-14, 16-21 and manufactured with the content of the specified ALS in the drug in an amount of 1.0 wt. The filler was a gel based on honey (15 parts), onion juice (3 parts), garlic juice (1 part) and burdock oil (1 part). The drug was used as a mask to strengthen the hairline. The gel was applied on the head for 2-3 hours. The procedure was repeated 1 time per week. The therapeutic effect was manifested after 5-12 sessions and was controlled by the state of the hairline.

 PRI me R 23. A drug based on ALS obtained in the same way as described in examples 1-6, 9-14, 16-21 was made with the content of the specified ALS in the drug in an amount of 0.1 wt. The filler was lanolin in a mixture with spermaceti in a ratio of 1: 3. The drug was used as an ointment to restore the skin after dermatosis. The use of the drug contributed to the acceleration of recovery processes by 2-5 times compared with the control.

 PRI me R 24. A drug based on ALS obtained in the same way as described in examples 1-6, 9-14, 16-21 was made with the content of the specified ALS in the drug in an amount of 0.1 wt. The filler was cocoa butter. The drug was used in the form of suppositories for the treatment of endometritis. The therapeutic effect when using the drug was manifested on days 14-21, which was monitored according to the indicators of the inflammatory reaction, histological studies, blood analysis and secretions.

 PRI me R 25. A drug based on ALS obtained in the same way as described in examples 1-6, 9-14, 16-21 was made with the content of the specified ALS in the drug in an amount of 1.0 wt. The filler was a mixture of lanolin with petroleum jelly in a ratio of 1: 3. The drug was used in the form of an ointment for the treatment of mucosal erosion and contributed to the acceleration of recovery processes 3-5 times in comparison with the control.

PRI me R 26. Amino acid analysis of the composition of ALS obtained in the same way as in examples 1-18, was carried out in three directions, which were determined:
total amino acid composition;
amino acid composition of peptides and free amino acids in the absence of acid-insoluble proteins;
composition of free amino acids.

Total amino acid analysis of BAS, obtained as in Examples 1-18, was performed by means of hydrolysis in 6 N hydrochloric acid (6 N HCl) at ¹¹⁰ ° C for 24 hours under vacuum. At the end of hydrolysis, the samples were evaporated in vacuo over dry sodium hydroxide (NaOH). The dry residue was diluted in bicarbonate (0.2 N) sodium citrate buffer solution, pH 2.2, and applied to the ion exchange columns of amino acid analyzers.

 To remove acid-insoluble proteins, 0.5 ml of 3% sulfosalicylic acid was added to 0.5 ml of the sample, left for 1 h during which the composition was periodically stirred. The denatured proteins were removed by centrifugation at an acceleration of 8000 g for 15-20 minutes. The resulting supernatant was subjected to general amino acid analysis.

 The content of free amino acids in BAS, obtained in the same way as in examples 1-18, was determined after equilibration of the samples with a bicancinormal (0.2 N) sodium citrate buffer solution at pH 2.2 and after filtration through a paper desalted filter was applied to ion-exchange columns of analyzers amino acids.

 The elution of amino acids from the columns was carried out with sodium citrate buffer solutions with a pH of 3.25; pH 4.25 and pH 5.28. Fractionated amino acids were treated with ninhydrin reagent and the optical density was determined on a flow photometer at a wavelength of 440 and 570 nm.

PRI me R 27. The content of total lipids in ALS obtained in the same way as in examples 1-18, was determined as follows. 0.5 ml of ALS was mixed with 1.5 ml of concentrated sulfuric acid (H ₂ SO ₄₎ and heated at 95 ^{° C} for 15 min. After cooling, 1.5 ml of Hemapol phosphovaniline reagent was added to 0.1 ml of the hydrolyzate according to the known method for determining total lipids by specific color (JMKnight, S. Anderson, JMRawle. Clinical Chemistry 1972, 18, p. 199). The optical density of the solution was determined at a wavelength of 530 nm.

PRI me R 28. The analysis of the nucleotide composition of ALS obtained in the same way as in examples 1-18, was carried out as follows. Acid-soluble purine and pyrimidine nucleotides were extracted from 1.5-5.5 ml of the sample with 5% cooled perchloric acid (HClO ₄ ). Samples after 20 min of cold extraction were centrifuged at an acceleration of 2500 g for 10 min, the precipitate was washed three times with 5% perchloric acid (HClO ₄ ), supernatants were combined and used to isolate acid-soluble nucleotides. Acid soluble fraction was hydrolyzed monomolar chloric acid (1 M HClO ₄₎ at ¹⁰⁰ ° C for 1 hour to piimidinovyh monophosphates and purine bases. The solutions were neutralized with potassium hydroxide (KOH) and the hydrolysis products were separated on a DAUEX 50x4 cation exchanger using stepwise elution: distillate (Н ₂ О) for uric acid and uridine monophosphate (UMF): two-decimolar perchloric acid (0.2 M HClO ₄ ) for xanthine and monophosphate (CMF); tetdecimolar perchloric acid (0.4 M HClO ₄ ) for hypoxanthine; unimolar perchloric acid (1.0 M HClO ₄ ) for guanine; bipolar perchloric acid (2.0 M HClO ₄ ) for adenine.

 Nucleotides were identified by chromatographic mobility on a cation exchanger in comparison with standards, as well as by absorption spectra in the region of 200-300 nm. The number of nucleotides was determined using molar extinction coefficients.

 PRI me R 29. The immunomodulatory activity (properties) of ALS obtained in the same way as in examples 1-18, was determined by the activation of macrophages of peritoneal exudate of Wister rats in vitro, by enhancing the enzymatic activity in the restoration of nitro-blue tetrazolium in diformazan (HCT test) and in enhancing phagocytic activity to E. coli.

20 ml of a colorless Hanks solution containing 20 u / ml heparin was injected into the abdominal cavity of decaritated animals using a needle with a syringe. After 2-3 minutes of abdominal massage, an incision was made and the injected liquid was aspirated from the cavity with the same syringe, which after filtration through a nylon filter was centrifuged at an acceleration of 150-200 g for 15 minutes. The resulting precipitate was suspended in half the volume of a fresh portion of Hanks solution and centrifuged again under the same conditions. Washing of the isolated cells was carried out 3-4 times, after which the concentration of cells in the suspension was adjusted to 80x10 ⁶ cells / ml. The content of macrophages in the tissue exudate of all cells was 25-35%
The enzymatic activity of macrophages of rat peritoneal exudate was determined spectrophotometrically to restore nitro-blue tetrazolium to formazin. The precipitate of rat peritoneal exudate cells was diluted with a colorless Hanks solution to a concentration of 80x10 ⁶ cells / ml. To 50 μl of the cell suspension, 50 μl of a sample with a pH of 7.4 and a peptide concentration of 1.0 mg / ml was added. The mixture was incubated at a tempertature ^{37 °} C water bath with constant shaking for 30 min, after which was added 50 .mu.l of saturated ^at 20 ° C a solution of nitro-blue tetrazolium firm Reanal (Hungary) and incubation was continued for another 30 min. Then 3 ml of acetone was added and centrifuged at an acceleration of 2000 g for 20 minutes. The supernatant acetone extract was photometric at a wavelength of 515 nm.

 As a control of comparison of enzymatic activity, a test with a preservative in physiological saline was used.

 The phagocytic activity of rat peritoneal exudate cells was determined by their ability to capture E. coli.

To 50 μl of the cell suspension was added 50 μl of a sample with a pH of 7.4 and a peptide concentration of 0.05 mg / ml. This mixture was incubated at ³⁷ ° C in water bath with permanent shaking for 30 minutes. After incubation the mixture was suspended in 10 ml colorless Hank's solution at ⁴ ° C and centrifuged at 150-200 g for 15 min. The precipitate of rat peritoneal exudate cells was diluted with Hanks solution to a concentration of 2.5x10 ⁶ cells / ml. The resulting slurry in an amount of 0.2 ml was applied to a glass measuring 18x18 mm and incubated at ³⁷ ° C for 30 minutes in an atmosphere with 5% -th carbon dioxide (CO ₂₎ for attachment to glass macrophages.

After incubation, the glass was rinsed 3 times in beakers with Hanks medium to remove non-adherent cells. 0.2 ml of E. coli suspension was added to the remaining cells on the glass at a concentration of 20x10 ⁶ cells / ml and incubated for 45 minutes under the same conditions. Subsequently, the glass was again rinsed three times, fixed with methanol and stained according to Romanovsky-Giemsa.

The phagocytosis index (IF) was determined by the formula:
IF (0 + 2.5n + 6T + 10R) / number of cells (1) where 0 is the number of cells that are not phagocytic in E. coli;
n the number of cells that have absorbed 1-4 cells;
T is the number of cells that have absorbed 5-7 cells;
p is the number of cells that have absorbed 8-9 cells;
R is the number of cells that have absorbed 10 or more E. coli cells. Identification of macrophages was performed by staining of nonspecific esterase.

 PRI me R 30. The immunomodulatory activity (properties) of ALS obtained in the same way as in examples 1-18, was determined by the activation of neutrophils of blood of Wistar rats in vitro experiments to enhance the enzymatic activity in the restoration of nitro-blue tetrazolium in formazan (HCT test) and the enhancement of phagocytic activity to E. coli.

 Blood from rat caudal artery was immediately collected into test tubes with a colorless Hanks solution on heparin and mixed. To the resulting mixture was added 6% T-500 dextran in a 3: 1 ratio and the tubes were refrigerated for one hour. After erythrocyte sedimentation, the supernatant of leukocytes was pipetted and centrifuged at an acceleration of 250 g for 15 minutes. Red blood cells were removed from the leukocyte mass by hemolysis by suspending the leukocyte pellet in distilled water, then a Hanks solution was added and the mixture was centrifuged. The white blood cell sediment was resuspended in fresh Hanks solution and centrifuged under the same conditions. The leukocyte washing operation was carried out 3 times.

 Determination of the enzymatic activity of blood neutrophils was performed spectrophotometrically to restore nitro-blue tetrazolium to formazan.

The rat blood leukocyte sediment was diluted with a colorless Hanks solution to a concentration of 80x10 ⁶ cells / ml. To 100 μl of the cell suspension was added 300 μl of BAS obtained according to examples 1-26, the pH of which was adjusted to 7.4 and the concentration of peptides to 1.0 mg / ml. This mixture was incubated at ³⁷ ° C in water bath with permanent shaking for 30 minutes after which was added 400 .mu.l of a solution of nitro-blue tetrazolium firm Reanal (Hungary) and further operations were carried out in the same sequence as that in Example 29.

 The phagocytic activity of rat blood leukocytes was determined by their ability to capture E. coli.

To 50 μl of a leukocyte suspension, 50 μl of ALS with a pH of 7.4 and a peptide concentration of 0.05 mg / ml was added. The mixture was incubated, and then added 10 ml of a colorless Hank's solution at ⁴ ° C and centrifuged at 150-200 g for 15 min. Leukocyte pellet was diluted in 0.3 ml Hanks solution and added 0.05 ml E.coli suspension at a concentration 500h10 ⁶ cells / ml and then incubated for 30 min at 37 ^C. After incubation, smears made in which the amount of phagocytic after dyeing cells and phagocytosis index (IF) were calculated as described in example 29.

 PRI me R 31. The immunomodulatory activity (properties) of ALS obtained in the same way as in examples 1-18, was determined in experiments in vivo in mice of the BALB / C line. The animals were given two subcutaneous injections of the drug with an interval of two days at a dose of 0.5 ml / kg of animal weight at a peptide concentration of 1.0 mg / ml. The analysis of immunomodulatory properties was performed two days after the last injection. In each experiment, at least 10 animals were used. The control was animals treated according to the indicated scheme with a 0.08% preservative dissolved in physiological saline.

Phytohemagglutinin (PHA), and E. coli lipopolysaccharide (LPS) was used as a B-cell mitogen to determine the reaction of blast transformation of lymphocytes as a T-cell mitogen. Lymphocytes isolated from spleen, were incubated with mitogens data for 72 hours at ³⁷ ° C in an atmosphere with 5% -th carbon dioxide (CO _2). The level of cell proliferation was evaluated radiometrically by incorporation of ³ H-thymidine. For this, ³ H-thymidine (3.7 × 10 ⁴ Bq / sample) was added to the cultures and incubated for 3 hours. Then the samples were applied to millipore filters, washed three times with 199 medium, then with a 5% solution of trichloroacetic acid (TCA) and ethanol . The filters were dried, placed in bottles with ZhS-106 scintillation liquid, and the radioactivity of the samples was determined using a Mark-111 counter.

 PRI me R 32. The immunomodulatory efficacy (properties) of ALS obtained in the same way as in examples 1-18, was determined by stimulation of the natural killer activity of splenocytes of mice in experiments in vivo. The animals were given two subcutaneous injections of the drug for two days at a dose of 0.05 ml / kg of the animal's weight at a concentration of peptides of 1.0 mg / ml in them. An analysis of immunomodulatory properties was performed the day after the last injection. In each experiment, at least 10 animals were used. The control was animals treated according to the indicated scheme with a 0.08% preservative dissolved in physiological saline.

× 100,(2) где максимальное высвобождение обозначает радиоактивность, при которой все клетки лизированы. В качестве контроля для определения усиления естественной киллерной активности предлагаемыми БАС применяли консервант, растворенный в физиологическом растворе.The state of the natural killer activity of splenocytes was determined on lymphoma cells of YAC-12 mice labeled with the ⁵¹ Cr chromium isotope. The incubation was carried out for 18 h at a ratio of target cells and effector cells of 1:25 (1x10 ⁵ : 2.5x10 ⁶ cells). The specific release of the chromium isotope ⁵¹ Cr, corresponding to the cytotoxic activity of effector cells, was calculated by the following formula:
⁵¹ Cr

× 100, (2) where maximum release refers to radioactivity in which all cells are lysed. As a control, to determine the enhancement of natural killer activity, the proposed ALS used a preservative dissolved in physiological saline.

 PRI me R 33. Immunomodulating effectiveness (properties) of ALS obtained in the same way as in examples 1-18, was determined by the functional activity of the thymus of mice in experiments in vivo under conditions similar to example 31.

 The effect was evaluated by the titer of thymic serum factor (TSF). The method is based on the ability of thymus hormones to restore the sensitivity of spontaneous rosette-forming spleen cells of adult thimectomized mice to antithymocyte serum.

 The blood serum of the experimental mice was passed through an Amicon CENTRIFLO CF-50A system ultrafilter. 0.1 ml of whole and serially diluted 199 serum was used in the reaction. The control was Wednesday 199.

The spleen was taken from mice 10-14 days after thimectomy. Cells were extracted from it by dissolving the tissue with preparative needles in medium 199. After filtering through a nylon sieve, they were washed twice with medium 199 by centrifugation at an acceleration of 4000 g. The pellet was resuspended in medium to a concentration of 4x10 ⁷ cells / ml, and thereby a slurry in a volume of 0.1 ml, which was added to tubes containing serum and the medium 199. The tubes are mixed by pipetting and incubated at ³⁷ ° C for 60 min.

After incubation, 0.1 ml of anti-thymocyte serum and complement of guinea pig in a 1:10 dilution were added to all tubes. The suspension was incubated at 37 ° C for another 30 minutes. Then, 0.1 ml of a sheep erythrocyte suspension (12 x 107 cells / ml) was added to the mixture, mixed, centrifuged at 250 g for 5 minutes and incubated for 60 minutes at 4-8 ° C. Test tube pellets were resuspended for 5 minutes. Counting rosettes was carried out in the camera Goryaeva. A lymphocyte that connected four or more red blood cells was taken for the outlet. The TSF titer was considered the last dilution of serum, causing a 50% reduction in the number of rosette-forming cells (ROCK) in relation to the control. The results were expressed as the logarithm of the titer at base 2, that is, in the form of log A, where A is the titer value. complement guinea pig in a 1:10 dilution. The suspension was incubated at a temperature of 37 ^about With another 30 minutes To the mixture was added 0.1 ml of a suspension of sheep red blood cells (12x10 ⁷ cells / ml), stirred, centrifuged at 250 g for 5 minutes and incubated for 60 minutes at 4-8 ^C. The pellet was resuspended in 5 min Counting rosettes was carried out in the camera Goryaeva. A lymphocyte that connected four or more red blood cells was taken for the outlet. The TSF titer was considered the last dilution of serum, causing a 50% reduction in the number of rosette-forming cells (ROCK) in relation to the control. The results were expressed as the logarithm of the titer at base 2, that is, in the form of log A, where A is the titer value.

PRI me R 34. The effectiveness of ALS obtained in the same way as in examples 1-18, which manifests itself in enhancing the repair ability of rat liver hepatocytes in vivo after subcutaneous injection of carbon tetrachloride (CCl ₄ ), was determined by restoring the activity of Na, K-ATPase as follows.

Male rats weighing 230-250 g of the Wistar line were injected subcutaneously daily for 4 days with carbon tetrachloride (CCl ₄ ) in a mixture with vegetable oil in a ratio of 1: 1 at a dose of 4.0 ml / kg of animal weight. 4 hours after the first injection of carbon tetrachloride (CCl ₄ ), the experimental animals received the first of 4 subcutaneous injections of the agent with an interval of one day at a dose of 0.05 ml / kg of animal body weight. The effectiveness of the drug was recorded the day after the last injection. Moreover, as a control of comparison, we used the liver of animals similarly affected by carbon tetrachloride (CCl ₄ ), but without the use of the studied ALS, as well as the liver of intact animals.

To determine the activity of Na, K-ATPase, hepatocyte cell membrane fractions were used. To this end, 15 g of the chilled liver of three rats was homogenized in 150 g of a one millimolar borate buffer solution (1 mM Na ₂ B ₄ O ₇ ) with a pH of 7.5 in the presence of a five-decimillimolar calcium chloride solution (0.5 mM CaCl ₂ ) using a manual glass homogenizer with teflon pestle. The homogenized mixture was centrifuged at 150 g at ⁴ ° C for 10-12 min. The supernatant was subjected to repeated centrifugation at an acceleration of 100,000 g for 20 minutes, and the resulting precipitate was washed 3 times in 130 ml of buffer under the same centrifugation conditions. At the end of washing, the precipitate was resuspended in the buffer solution, adjusting the protein concentration according to the SDS Lowry method to 3.0 mg / ml, which was used in the analyzes.

 All operations to obtain the membrane fraction were performed in the cold.

The activity of Na, K-ATPase activity of hepatocyte plasmatic membranes was determined in a standard medium consisting of: shestidesyatishestimillimolyarnogo sodium chloride (66 mM NaCl), tridtsatichety- rehmillimolyarnogo potassium chloride (34 mM KCl), pyatimillimolyarnogo magnesium chloride (5 mM MgCl _2), dvadtsatipyatimillimolyarnogo (25 mM) Tris-HCl, to which, before the start of the experiment, five-millimolar adenosine triphosphoric acid (5 mM ATP) was added.

To 1.8 ml of the standard medium was added 0.2 ml of a suspension of membrane fraction with protein concentration of 3.0 mg / ml and incubated at ³⁷ ° C for 15 minutes, after which the reaction was stopped by adding 0.6 ml of 50% Trichloroacetic acid (TCA) solution. The mixture was centrifuged at an acceleration of 300-500 g for 15 minutes. To 1.0 ml of the obtained supernatant was added 4 ml of a two-decimolar (0.2 M) acetate buffer solution with a pH of 4.0. 0.5 ml of a 1% solution of molybdatammonium in a 1% solution of sulfuric acid (H ₂ SO ₄ ), 0.5 ml of ascorbic acid in a 0.16% solution of copper sulfate (CuSO ₄ ) and, after stirring, were incubated 10 min at 25 ^C. The optical density was determined spectro- metrically at a wavelength of 700 nm.

 The activity of Na, K-ATPase was calculated by the difference between the activities of total ATPase and Mg-ATPase. To determine the activity of Mg-ATPase, similar studies were carried out, however, instead of 0.6 ml of trichloroacetic acid (TCA), 0.3 ml of a 0.05% strophanthin solution was added to the mixture.

It was found that with toxic damage to the liver by carbon tetrachloride (CCl ₄ ), the activity of Na, K-ATPase decreases by 3.5 times, however, when using the proposed ALS, as in example 11, its activity is restored in the experiment by no less than 50% by compared to the norm.

PRI me R 35. The effectiveness of ALS obtained in the same way as in examples 1-18, which is manifested in enhancing the repair ability of rat liver hepatocytes (in vivo) after subcutaneous injection of carbon tetrachloride (CCl ₄ ), was determined by the restoration of activity in blood aminotransferases and potassium (K +) concentrations in rat liver hematocytes as follows.

 In animals treated as in example 34, blood aminotransferase activity was checked using a Hemapol test system according to a specific colorimetric photometric method (S. Reitman, S. Frankel. American Journal of Clicical Pathology. 1957-28-56) .

0.05 ml was added to 0.25 ml of a solution containing a decimolar solution (0.1 M) of L-aspartate, two-millimolar (0.002 M) 2-oxoglutarate and decimolar (0.1 M) phosphate buffer solution rat blood serum and incubated at a temperature of 37 ^about C for 60 minutes At the end of the incubation, 0.25 ml of millimolar (0.001 M) 2,4-dinitrophenylhydrazine dissolved in unimolar hydrochloric acid (1 M HCl) was added to the solution and the sample was left for 20 min at room temperature, after which 2.5 ml of four decimolar (0.4 M) sodium hydroxide (NaOH) was stirred and after 10 minutes the absorbance was measured at a wavelength of 520 nm. Instead of 0.05 ml of blood serum, 0.05 ml of physiological saline was added to the control solution.

Toxic liver damage with carbon tetrachloride (CCl ₄ ) is accompanied by the destruction of hepatocytes and, accordingly, the release of aminotransferases, and therefore their activity in the blood serum increases by 2 times. At the same time, the use of the proposed BAS, as in example 11, allows you to restore in the experiment such activity is not less than 59% compared with the norm.

The concentration of potassium (K ⁺ ) in the hepatocytes of the liver of experimental rats was determined by the standard method of flame photometry, according to Bricker, on a flame photometer PFM-1 (Brikker V.N. Determination of K, Na by flame photometry. Laboratory, 1961, 7, p. .3-6).

It was found that with toxic damage to the liver by carbon tetrachloride (CCl ₄ ), the activity of Na, K-ATPase decreases by 3.5 times, which is accompanied by a 40% decrease in the concentration of K ⁺ in hepatocytes of rat liver. At the same time, the use of ALS obtained in the same way as in example 11, allows you to restore the concentration of K ⁺ not less than 43%
The invention is not limited to the specific embodiments set forth. Various options may be practiced within the spirit and scope of the appended claims.

 Identification of the proposed UAS among other biologically active agents is possible directly by biochemical analysis of the composition and indirectly by the magnitude of the immunomodulating effect that UAS exhibits.

It was found that it is the content of the modified components of morphoplasm and glycocalyx of cells with a modified structure in the composition of ALS that provides a significant increase in the body's immune response compared to the control. This allows us to identify UAS by the following set of indicators:
a) to increase the activity of macrophages of rat peritoneal exudate, namely to increase the enzymatic activity of macrophages controlled by the NBT test by not less than 54% (when using the nitro-blue tetrosolium HCT in the experiments of the Hungarian company Reanal, and to increase phagocytic activity E.coli no less than 39%
b) to increase the activation of rat blood neutrophils, namely, to increase the enzymatic activity controlled by the NBT test by not less than 67% (when using the nitro-blue tetrosolium HCT in the experiments of the Hungarian company Reanal) and to increase the phagocytic activity of E. coli no less than 44%
c) to enhance the reaction of blast transformation of T-lymphocytes by not less than 42% and B-lymphocytes by not less than 50%
d) to increase the functional activity of the thymus according to the content in the blood of rats of thymic serum factor (TSF) by not less than 167%
e) to increase the natural killer activity of rat splenocytes by not less than 66%
e) by increasing the ability to stabilize the plasma membranes of rat liver hepatocytes after administration of carbon tetrachloride (CCl ₄ ):
to restore the activity of Na, K-ATPase by at least 24%
to restore transferase activity in rat blood by not less than 22%
to restore the ratio of potassium (K) and sodium (Na) in rat liver hepatocytes by not less than 13.8%
At the same time, control was used as a comparison base in the form of a preservative introduced into the body, dissolved in physiological saline.

 The system of indicators of immunomodulating and regenerative effects (properties) of the proposed BAS allows us to identify it by the manifested biological activity, namely, as an immunomodulator.

 For ALS and a drug based on it, there is a tendency to manifest an effect of stimulating the protective functions of the body’s immune system, aimed both at eliminating internal pathology (stimulating the activity of natural killers) and increasing the body’s overall resistance to infectious diseases, which is significantly higher than similar indicators in the above known ALS.

The immunomodulating properties of various biologically active agents have been experimentally determined in accordance with a set of indicators identifying the studied objects. The experimental results are summarized in table 6, which shows the quantitative indicators of the effects of known BAS N ₁ , N ₂ , N ₃ obtained by the technology described in EP-A-02499563, SU-A-139404 and US-A-4455302, respectively. and the proposed UAS, presented in the form of groups WS, TS and LM, namely, summarized by the results:
WS examples 1-4 (components of morphoplasm and glycocalyx cells);
TS examples 7-9 (thermostable components of morphoplasm and glycocalyx cells);
LM examples 11-13 (low molecular weight components of morphoplasm and glycocalyx of cells M ≅ 10.0 kDa).

 As a control object, a physiological solution of sodium chloride containing a preservative was used. The results are summarized in table. 6 and comparatively characterize the proposed and known ALS. The above results relate to the characteristics of the proposed UAS obtained in the form of a mixture of an unknown structure and the data necessary for its identification by the effect that determines its purpose.

 The analysis of the data in Table 6, along with the ability to identify the proposed tool, also allows you to evaluate the specific result provided by the proposed BAS, which consists in a significant improvement in the properties of biological activity due to the presence of hydrolysis products containing modified components of morphoplasm and glycocalyx of cells to a modified structure, the effect of which in known ALS not detected.

 The properties of ALS and a drug based on it are closely dependent on its composition.

 As shown by the results of biochemical analysis (Table 7), BAS contains mainly organic compounds such as polypeptides, peptides, amino acids, bound and free carbohydrates, lipids, nucleotides and other compounds. The data in Table 7 show the mass distribution of organic compounds in the composition of ALS when using cattle embryonic tissue as a raw material. In addition, Table 7 shows the results of obtaining various ALS groups (WS, TS and LM) sequentially by all the proposed methods. In addition to the results of the method that provides obtaining ALS-WC and in the implementation of which animal tissue is used as raw material, taken at the stage of the process of modifying the components of morphoplasm and glycocalyx of cells with the above processes, table 7 also includes the results of the method for obtaining the claimed ALS in the form thermostable components of BAS-ST and in the form of low molecular weight components of BAS-LM.

 The data in Table 7 allow us to carry out a comparative analysis of the results of the proposed and known methods for producing ALS, namely, a comparative analysis taking into account the fact that of the known ALS N3 (US-A-4455302), ALS N2 (SU-A-139404) and ALS N1 (EP-A-0249563) are the best in their characteristics.

 A comparative analysis of the specific activity of the proposed and known ALS shows (Tables 6 and 7) that their specific activity depends on the composition of the ingredients, the ratio of which significantly affects the specific activity.

 A characteristic feature of the biochemical composition of ALS is the presence of carbohydrate-containing components in it, the modification of which to a large extent determines its properties.

 In this case, the carbohydrate content in the most active fractions (fractions 3.4 and 5, Table 4) of low molecular weight components with a molecular weight of M 0.8-10.0 kDa (oligocarbohydrates, glycopeptides, glycolipids, nucleotides) are crucial in identifying and characterizing the proposed BASS.

 Thus, a comparative analysis of the specific activity and composition of the ingredients (Tables 6 and 7) shows that a higher activity of ALS in comparison with analogues correlates with a higher content of carbohydrates in the composition of components with a molecular mass of 0.8-10.0 kDa. For the proposed tool, these values correspond to the values: 6.8 ± 3.5% 7.5 ± 3.5% and 8.2 ± 3.3%, whereas for BAS analogues N1, N1 and N3 they are: not more than 0, 2% 2.7 ± 1.1% and 3.0 ± 0.5%, respectively. To an even greater extent, this dependence correlates with the specific content of carbohydrates in relation to other organic compounds of this fraction.

(3) где С_у, С_п концентрации соответственно углеводов и пептидов в компонентах с молекулярной массой М 0,8-10,0 кДа.In this regard, a significant indicator is the ratio of the content of carbohydrates and peptides in components with a molecular mass of 0.8-10 kDa M (Tables 2 and 7), which is determined by the formula: 3
G

(3) where C _y , C _{p are the} concentrations of carbohydrates and peptides, respectively, in components with a molecular weight of M of 0.8-10.0 kDa.

 For the proposed UAS, the value of this indicator lies in the range of G 0.25 ÷ 0.60, while for known UAS the value of this indicator does not exceed the value of G <0.25.

 Thus, the value of the indicator G generally characterizes the property of hydrolysis products and depends on the raw materials used, on the totality of the operations of the processing method for the specified raw materials and the modes of hydrolysis processes and is one of the characteristic indicators of the biochemical identification of the proposed BAS.

 The totality of the characteristics of a biologically active agent and the totality of the characteristics of the method for its preparation and the drug based on this agent are causally related to the specific activity of ALS.

 The inventions relating to a biologically active agent, a method for its preparation and a medicinal product based on ALS are based on the choice of feedstock, the choice of the conditions for the process of obtaining ALS and a drug with guaranteed high specific activity. Moreover, the selected technology for producing ALS provides reproducible results, that is, it allows to standardize the receipt of ALS with desired properties, namely with a given fixed immunomodulating activity that determines the pharmacological, therapeutic and general biological effects when using the invention.

 The resulting target products in general and particular cases of the implementation of the method for producing ALS are equivalent in nature to their biological effects, but differ in their specific activity. Therefore, the feasibility of using one or another specific ALS is determined by the requirements of the object for which they are intended, that is, the requirements of veterinary medicine and / or medicine. In this regard, in each specific case of using the target product, the choice of a particular ALS variant is determined by the ratio of the requirements for its immunomodulating activity and overall effectiveness with the cost (economy) of its production. In particular, the high efficiency of the agent containing low molecular weight components with a molecular mass of M ≅ 10.0 kDa of the proposed BAS-LM is fully manifested in an increase in the activity of macrophages, neutrophils, lymphocytes, and natural killers, which ensures an increase in regenerative processes (Table 6) . For farm animals, such effects are accompanied by an increase in the additional weight gain of the animals during intensive feeding (Table 8). In the treatment of respiratory and infectious diseases (parainfluenza), this is mainly not so significant, and it is economically feasible to use an agent containing water-soluble components of morphoplasm and glycocalyx of BAS-WS cells. In the treatment of inflammatory processes (mastitis), it is more advantageous to use an agent containing thermostable components of BAS-ST.

 When preparing preparations for external use in the form of ointments, suppositories, gels, compresses, intranasally or for feeding animals instead of BAS-LM, it is economically feasible to use BAS containing thermostable components of BAS-ST.

 At the same time, to obtain the maximum effect, it is more expedient to use a tool to stimulate the protective functions of the body’s immune system, aimed both at eliminating internal pathology (stimulating the activity of natural killers) and at increasing the body’s overall resistance to infectious diseases, which is significantly higher than those considered above known ALS.

 The use of ALS in infectious diseases of young cattle (parainfluenza, acute respiratory infections in calves and others) ensured the recovery of animals in 82-93% of cases. At the same time, when using known agents under the same conditions, recovery of animals was observed in no more than 70% of cases, and with spontaneous cure in the control group within 58-67% of cases.

 The use of the drug gave positive results when introduced into the body of animals in the treatment of inflammatory processes.

The use of ALS for prophylactic purposes reduced the level of calf disease by parainfluenza to 10%, while using known agents, the incidence rate of animals reached 12-17% and in the control group 22-28%
The use of the drug increased the resistance of animals to infectious diseases, thereby reducing the death rate of the livestock.

 Normalization of the general biological state of farm animals by the preparation contributed to the improvement of production indices and, in particular, to the restoration of additional weight gain without loss of meat quality, bringing these indices closer to the genetically programmed level during intensive feeding. This feature favorably distinguishes the proposed UAS and preparations based on it from hormonal and other growth stimulants, which, by increasing quantitative indicators, adversely affect animal homeostasis and reduce the quality of products.

The introduction of ALS and preparations based on it into the body of an animal having deviations of the parameters of homeostasis from the norm, provided intensive feeding increased an additional gain by an average of 37-100% from the gain in the control group not treated with the proposed ALS. At the same time, when using known BAS analogues (N1, 2, and 3) under the same conditions, the additional gain in comparison with the gain in the control group averaged 12-24%
Normalization of homeostasis parameters also contributes to an increase in: milk yield of cattle and small cattle; egg production of poultry; qualities of fur of fur animals; survival of young animals; honey collecting bees and the like.

 ALS and a drug based on it are non-toxic, did not cause side effects (allergies and the like).

 Comparative data on the pharmacological, therapeutic and general biological effect of the use of the proposed ALS and the preparation based on it in animals are given in Table 8. An analysis of the data in this table shows that the biological activity of ALS significantly exceeds the biological activity of analogues.

 Thus, the proposed ALS and a drug based on it have a wide range of applications in veterinary medicine and medicine and can be used to normalize the physiological organism in the treatment of diseases that are controlled within the competence of the body’s immune system.

 According to the invention, the agent and the preparation based on it are used as follows. The parameters of homeostasis are determined, then, if they deviate from the norm, the immune system is stimulated by the introduction of ALS until the normalization of the indicated parameters of homeostasis.

 The indicated stimulation of the immune system is carried out by activating the reticuloendothelial system, T- and B-systems, neutrophils and / or the natural killer population of the body, which provides increased activity of macrophages of peritoneal exudate and blood neutrophils in vitro, increased stimulation of the in vivo response of lymphocyte blast transformation , increased stimulation of the functional activity of the thymus, increased stimulation of the natural killer activity of splenocytes in vivo, as well as increased reparative abilities of hepatocytes in the liver after administration of agents or partial hepatectomy and so on.

To this end, the method of introducing into the body of ALS and a drug based on it selects a number of them:
intramuscular injections at a dose of 0.005 0.5 ml / kg body weight with an interval between injections of 1-7 days;
inhalations in a dose of 0.012-0.12 ml / kg body weight with an interval between inhalations of 4-48 hours; sublingually at a dose of 0.012-0.12 ml / kg body weight with an interval between administrations of 4-48 hours;
drinking at a dose of 0.02-1.0 ml / kg body weight with an interval between drinking a 1-10 days;
integrally in pills at a dose of 0.02-1.0 ml / kg body weight with an interval of 1-10 days;
intranasally in a daily dose of 0.001-0.05 ml / kg body weight with an interval between instillations of 2-24 hours;
applications on the mucous or wound surface in the form of compresses, suppositories, powders, ointments, as well as gels;
and combinations thereof, to normalize the parameters of homeostasis.

 Information confirming the effectiveness of using the method of using BAS in the form of thermostable components of BAS-ST and a preparation based on it, in which physiological solution of sodium chloride was used as a filler, are presented in Tables 9-12. The tables show the dependence of the effect of normalizing the parameters of homeostasis on the above dose limits and the method of using the drug, which was controlled by the weight gain of the calves, which initially had a deviation from the norm in the parameters of homeostasis. In the table. 9-12 presents the results of the effectiveness of the drug with intramuscular injections with an interval of 3 days (table 9); with inhalation with an interval of 24 hours (table. 10); drinking out with an interval of 7 days (table 11); with intranasal use with an interval of 12 hours (table 12).

 Introduction to the body of ALS in the form of the proposed drug can stimulate: the cure of infectious diseases (parainfluenza, acute respiratory infections); cure for inflammatory diseases (mastitis, pneumonia, endometritis); increasing the resistance of animals to infectious and inflammatory processes; improving the general biological state of the body; increase in animal gain; functional activity of the liver tissue with hepatitis; liver regeneration after partial hepatectomy; healing of ulcers, wounds, skin during wounds and injuries; restoration of the blood formula with blood loss.

 The absence of side effects when using ALS and a drug based on it and data on the pharmacological, therapeutic and general biological effects on the animal’s body have determined the possibility of clinical testing of ALS and a drug based on it.

 When examining a group of patients, a positive effect on the physiological state of the body was determined, as well as an almost complete analogy to experimental preclinical studies in animals.

 A method of obtaining a biologically active agent can be implemented both in industrial and in laboratory conditions.

Claims (22)

 1. A biologically active agent with an immunomodulating property based on water-soluble hydrolysis products of ingredients of ground animal tissue represented by organic compounds, characterized in that it contains thermostable, low molecular weight components of morphoplasm and glycocalyx of cells modified by embryogenesis in the hydrolysis products and / or cell proliferation and / or differentiation and / or pathology preceding and / or concomitant tissue regeneration and / or repair and. 2. The tool according to p. 1, characterized in that it contains in the composition of the modified components of morphoplasm and glycocalyx of cells thermostable at T _n ≅ 120 ^o C components.  3. The tool according to p. 2, characterized in that it contains thermostable components in an amount of 30 to 100 wt.  4. The tool according to claim 1, characterized in that it contains low molecular weight components with a molecular mass of M ≅ 10.0 kDa as part of the modified components of the morphoplasm and glycocalyx of the cells.  5. The tool according to claim 2, characterized in that it contains low molecular weight components with a molecular mass of M ≅ 10.0 kDa as part of the modified components of the morphoplasm and glycocalyx of the cells.  6. The tool according to p. 4, characterized in that it contains low molecular weight components in an amount of 30 to 100 wt.  7. The tool according to p. 5, characterized in that it contains low molecular weight components in an amount of 30 to 100 wt.  8. The tool according to p. 1, or 2, or 3, or 4, or 5, or 6, or 7, characterized in that it contains organic compounds in the composition of hydrolysis with modified components of morphoplasm and glycocalyx of cells in the following ratio, wt. . Polypeptides Up to 26.0
Peptides 10.0 22.5
Amino acids 32.0 60.1
Carbohydrates 10.5 28.7
Lipids, nucleotides and other organic compounds 2.0 11.0
9. The tool according to p. 8, characterized in that it contains carbohydrates in the composition of organic compounds in the following ratio, wt. consisting of: high-molecular compounds having a molecular weight M _{in the y>} 10.0 kDa up to 2.0 molecular compounds having a molecular weight M _n of 0.8 _at 10.0 kDa 2.5 11.5 free monomers M _{y m} <0.8 kDa, 6.0 17.0
10. The compound according to claim. 1 or 2 or 3 or 4 or 5 or 6 or 7, characterized in that the ratio by weight of carbohydrate and peptide contents in the composition of the hydrolysis product with molecular weight M _n of 0.8 _at 10 , 0 kDa lies in the range of G 0.25 0.6.  11. The tool according to p. 1, or 2, or 3, or 4, or 5, or 6, characterized in that it contains in the composition of hydrolysis with modified components of morphoplasm and glycocalyx of cells organic compounds in the following ratio, wt. Polypeptides 16.3 24.7
Peptides 11.9 17.1
Amino acids 32.0 48.2
Carbohydrates 14.6 20.8
Lipids, nucleotides and other organic compounds 5.1 9.3
12. The tool according to p. 2, or 3, or 5, or 7, characterized in that it contains in the composition of hydrolysis with modified components of morphoplasm and glycocalyx of cells organic compounds in the following ratio, wt. Polypeptides 5.5 12.7
Peptides 13.6 21.0
Amino acids 37.1 52.9
Carbohydrates 16.1 25.1
Lipids, nucleotides and other organic compounds 5.3 11.0
13. The tool according to p. 4, or 5, or 6, or 7, characterized in that it contains in the composition of hydrolysis products with modified components of morphoplasm and glycocalyx of cells organic compounds in the following ratio, wt. Polypeptides Less Than 0.3
Peptides 14.9 22.5
Amino acids 43.5 60.1
Carbohydrates 15.5 28.7
Lipids, nucleotides and other organic compounds 5.0 9.2
14. A method of obtaining a biologically active agent having an immunomodulating property, in which the washed raw material is crushed, the hydrolysis of the ingredients is obtained, followed by separation of the hydrolysis products, their filtration and isolation of the supernatant as the target product, characterized in that the animal tissue is used as the raw material modified during embryogenesis and / or proliferation and / or cell differentiation processes and / or pathology preceding and / or concomitant reg neratsii and / or reparation of the tissue, and it is carried out before hydrolysis homogenization and purification from non-destroyed tissue elements with release supernatant containing ingredients with components morfoplazmy and glycocalyx of cells.  15. The method according to p. 14, characterized in that the raw materials use a tissue selected from: embryonic tissue, differentiating tissue of thymus thymocytes, spermatogonia and spermatocytes of the testes, placental tissue, intestinal epithelial tissue, bone marrow cells, regenerating skin tissue, tissue of the regenerating liver, tissue of the regenerating limbs of amphibians, pathologically altered tissue taken before the stage of regeneration, and their combination.  16. The method according to p. 14, characterized in that the cleaning of undamaged tissue elements after homogenization in physiological and / or buffer solution is carried out by filtration and / or centrifugation at an acceleration of 150 250 g for 10 15 minutes  17. The method according to p. 14, characterized in that the grinding of raw materials from animal tissue is carried out before homogenization and lead to damage to the integrity of the cells, followed by extraction of their contents by additional washing with filtration and / or centrifugation at an acceleration of 300-15000 g for 10 20 minutes with the release of extracted shredded tissue.  18. The method according to p. 14, or 15, or 16, or 17, characterized in that the composition of the ingredients subjected to hydrolysis, the components of the morphoplasm and glycocalyx of the cells is 30-60% of the total mass of organic compounds. 19. The method according to p. 14, characterized in that the hydrolysis is carried out at a temperature of T _p 4 45 ^o C.  20. The method, characterized in that the hydrolysis is carried out in the presence of a preservative. 21. The method according to p. 14, or 15, or 16, or 17 or 19, or 20, characterized in that the products of hydrolysis of the ingredients are subjected to heat treatment at a temperature of T _n 60 120 ^o C to completely denature the thermolabile compounds and filtered and / or centrifuged they are accelerated over 1500 g for 20 40 min with the release of a supernatant containing thermostable components as the target product.  22. The method according to p. 14, or 15, or 16, or 17, or 19, or 20, characterized in that the hydrolysis products of the ingredients are subjected to fractionation by dialysis through a semi-permeable film and / or ultrafiltration, and / or gel filtration to isolate the low molecular weight fraction components with a molecular weight of М≅ 10.0 kDa as the target product.  23. The method according to p. 21, characterized in that the products of hydrolysis of the ingredients are subjected to fractionation by dialysis through a semipermeable film and / or ultrafiltration, and / or gel filtration to isolate the fraction of low molecular weight components with a molecular weight of M≅ 10.0 kDa as the target product.  24. A preparation for normalizing the physiological state of the human and animal body, containing a biologically active agent based on the products of hydrolysis of animal tissue ingredients, and an excipient, characterized in that, as a biologically active agent, it contains thermostable, low molecular weight components of morphoplasm and glycocalyx of cells as a part of hydrolysis modified during embryogenesis and / or proliferation and / or cell differentiation processes and / or pathology preceding and / or concomitant s regeneration and / or tissue reperatsii when its content in the formulation in an amount of 0.001 85.0 wt. the rest is filler.  25. The drug according to p. 24, characterized in that the biologically active agent contains organic compounds in the ratio, wt. Polypeptides Up to 26.0
Peptides 10.0 22.5
Amino acids 32.0 60.1
Carbohydrates 10.5 28.7
Lipids, nucleotides and other organic compounds 2.0 11.0
26. The drug according to p. 25, characterized in that the biologically active agent contains organic compounds in the ratio, wt. Polypeptides 16.3 24.7
Peptides 11.9 17.1
Amino acids 32.0 48.2
Carbohydrates 14.6 20.8
Lipids, nucleotides and other organic compounds 5.1 9.3
27. The drug according to p. 25, characterized in that the biologically active agent contains organic compounds in the ratio, wt. Polypeptides 5.5 12.7
Peptides 13.6 21.0
Amino acids 37.1 52.9
Carbohydrates 16.1 25.1
Lipids, nucleotides and other organic compounds 5.3 11.0
28. The drug according to p. 25, characterized in that the biologically active agent contains organic compounds in the ratio, wt. Polypeptides Less Than 0.3
Peptides 14.9 22.5
Amino acids 43.5 60.1
Carbohydrates 15.5 28.7
Lipids, nucleotides and other organic compounds 5.0 9.2
29. The drug according to p. 25, or 26, or 27 or 28, characterized in that the filler contains a physiological solution of sodium chloride, and / or saline buffer solution, and / or carbohydrate compounds, and / or mineral, animal fats, vegetable or synthetic origin and mixtures thereof.  30. The drug according to p. 25, or 26, or 27, or 28, characterized in that it further comprises a preservative.  31. A method of normalizing the physiological state of humans and animals, comprising introducing into the body a preparation based on products of hydrolysis of animal tissue ingredients, characterized in that a composition of 0.001 to 85.0 wt. a biologically active agent containing, as part of hydrolysis products, morphoplasm and glycocalyx components of cells modified during embryogenesis and / or proliferation and / or differentiation of cells and / or pathology preceding and / or accompanying tissue regeneration and / or repair, and 15 0 99.999 wt. filler, and the introduction of the drug into the body is carried out by intramuscular injection of the said agent at a dose of 0.005 0.5 mg per 1 kg of body weight with an interval between injections of 1-7 days and / or inhalation at a dose of 0.012-0.12 mg of the agent per 1 kg of body weight body with an interval between inhalations of 4-48 hours, and / or sublingual administration at a dose of 0.012-0.12 mg of agent per 1 kg of body weight with an interval between injections of 4-48 hours, and / or drinking at a dose of 0.02-1, 0 mg of agent per 1 kg of body weight with an interval between drinking 1 10 days, and / or integrated administration in pills at a dose of 0.02 1.0 mg of agent per 1 kg of body weight with an interval between administrations of 1 10 days, and / or intranasal administration in a daily dose of 0.001 0.05 mg of the drug per 1 kg of body weight with an interval between administrations of 2 24 hours, and / or applications on the mucous or wound surface in in the form of compresses, suppositories, powders, ointments and gels with an interval between applications of 1 10 days, until normalization of the physiological state parameters.

Images