RU2123345C1 - Agent for disbacteriosis treatment - Google Patents

Abstract

FIELD: medicine, gastroenterology. SUBSTANCE: an agent has a lyophilized mixture of bifidobacteria biomass, lysozyme and sodium nucleinate at the following ratio of component per a single dose: bifidobacteria biomass 10⁵-10⁸ CFU (colony forming units); lysozyme 5-20 mg, and sodium nucleinate 50-250 mg. Obtained preparation has a complex biological effect. EFFECT: enhanced effectiveness of an agent, decreased treatment period, stable curative effect. 3 tbl, 3 ex

Description

 The invention relates to medicine and can be used in gastroenterology, pediatrics, gerontology, as well as in obstetrics and gynecology, oncology, surgery and other fields of clinical medicine.

 The use of bacterial and certain immunomodulating biological products for the treatment and prevention of dysbiosis and intestinal dysfunctions is known. The most widely used are eubiotics from representatives of the intestinal normoflora, for example, bifidumbacterin, primadofilus, lactobacterin, colibacterin, bificol, acipol, etc. [Dorofeychuk V.G., Beyer L.V., Sazonova N.E. et al. Treatment and prevention of dysbiosis in children. Methodical recommendations, M., 1986; Dysbacteriosis and eubiotics. Thes. doc. Vseros. scientific and practical. Conf., M., 1996], the principle of which is based on the phenomenon of microbial antagonism. Regulation of microbiocenosis of the mucous membranes is carried out due to the direct effect on the microflora of production strains of eubiotics by filling in the deficit of its corresponding representatives, crowding out pathogenic and excess of conditionally pathogenic microorganisms, as well as mediated mobilization of local immunity and metabolic processes by decay products of bacterial cells, for example, muramyl dipeptide [MDP V.M., Uchaykin V.F., Murashova A.O., Abramov N.A. Dysbiosis. Modern opportunities for prevention and treatment. M., 1995; Shcherbakov I.T. Pathomorphology of the mucous membrane of the gastrointestinal tract in acute bacterial, viral intestinal infections and chronic colitis: Dis. Doct. honey. Sciences, M., 1996]. Bacterial drugs in most cases provide clinical improvement. However, their disadvantage is the need for prolonged use to achieve a stable bacteriological effect and slow correction of local immunity, especially in debilitated patients. In addition, with significant violations of the immune system, some eubiotics themselves can cause infectious complications [Tyurin M.V. Lactic acid bacteria as potential causative agents of human infectious diseases // Antibiotics and microecology of humans and animals. M., 1988, p. 175 - 178], to promote the spread of antibiotic resistance, to oppress other representatives of the normobiocenosis [Lesnyak SV, Evtukhova L.N., Shimchuk L.F. About the properties of drugs from normal human microflora // Antibiotics and microecology of humans and animals. M., 1988, p. 136 - 140].

 In the treatment of dysbacteriosis and intestinal dysfunctions, immune preparations are also used, for example, directed immunoglobulins (lactoglobulin anti-lipoprotein, complex immune preparation CIP, etc.) [Soboleva SV Centered directed lactoglobulins (microbiological aspects of the development and clinical use of drugs): Dis. Doct. honey. Sciences, Rostov-on-Don, 1991; Ozernaya A.S., Korshunov V.M., Uchaykin V.F. The use of bacterial and immunoglobulin preparations for the treatment of acute intestinal infections in young children // Actual problems of chemotherapy of bacterial infections: Abstract. doc. All-Union conf., 1991, v. 3, p. 471]. Their effect is based on the specific activity of the antibodies contained in them (immunoglobulins of classes G and M) in relation to the corresponding microorganisms and stimulation of nonspecific resistance of the body. Their disadvantages include the high cost, the difficulty of obtaining environmentally friendly raw materials, as well as the relatively high reactogenicity in sensitized patients, associated with the risk of adverse effects on the mucous membranes of excess exogenous IgG class immunoglobulins and products of the rapid death of microorganisms. In addition, the use of immunoglobulin preparations requires additional administration of eubiotics [Ladodo KS, Scherbakova E.G. etc. The use of biologically active additives to baby food. Method. recommendations. M., 1988].

 To correct local immunity of the gastrointestinal tract (GIT), stimulate nonspecific resistance, improve the adhesive properties of bifidobacteria and lactobacilli with dysbiosis and intestinal dysfunctions, especially in first-year-old children who are breast-fed, lysozyme is used, which can be considered "ideal" , physiological immunomodulator. Lysozyme refers to the factors of natural defense of living organisms and has not only a mild immunomodulating effect, but also a wide range of other valuable general biological properties. It increases protifoinfection and antitoxic resistance, participates in the digestion and regulation of many of the most important body systems, has antimicrobial, anti-inflammatory, hyposensitizing, antianemic and many other effects [Bukharin O.V., Vasiliev N. V. Lysozyme and its role in biology and medicine . Tomsk, 1974 .; Shcherbakova E.G., Rastunova G.A. New in the mechanisms of action of lysozyme // New in transfusiology, 1995, no. 10, p. 51-60]. Chicken egg protein lysozyme (an injection drug and lysozyme hydrochloride) used in medical practice is similar in its antigenic structure to endogenous lysozyme and has the same mechanisms of excretion and regulation of the level in blood and tissues, which minimizes its side effect. Lysozyme is used for dysbacteriosis and intestinal dysfunctions to enrich baby food as an additional tool that increases the effect of bifidobacteria and lactobacilli, as well as immune lactoglobulins [Ladodo KS, Shcherbakova E.G. etc. The use of biologically active additives to baby food. Guidelines. M., 1988 .; Shcherbakova E. G. A systematic approach to the treatment and prevention of intestinal microecology disorders. // Successes of theoretical and clinical medicine. Mater. I scientific RMAPO Sessions, M., 1995, p. 76].

As a prototype, a means for treating and preventing dysbacteriosis and intestinal dysfunctions was taken - bifilis (VIGEL), which is a lyophilized mixture of living biomass of bifidobacteria (B. bifidum strain 1) and lysozyme in the following ratio of components per dose: biomass of bifidobacteria - 10 ⁶ -10 ⁸ colony forming units (CFU), lysozyme - 5 to 20 mg [patent RU 2071339 C1, cl. 6 A 61 K 35/74, 38/43; publ. 01/10/97, Bull. N 1].

 Biphilis provides a rapid improvement in clinical and bacteriological parameters in patients with dysbiosis and intestinal dysfunctions, an improvement in the morphological and functional state of the gastrointestinal tract and the general condition of patients; with prophylactic use, it significantly reduces the frequency of dysbacterioses of various genesis and increases the anti-infectious resistance of the body, especially in newborns and young children deprived of breast milk, in weakened patients [Shcherbakova E.G., Korovina N.A., Bolshakova A.M. and others. Integrated probiotic bifilis in the treatment and prevention of dysbiosis in children // Scientific and practical aspects of improving the quality of products of baby and herodietic nutrition .: Proceedings of the 1st Int. Conf., M., Pishchepromizdat, 1997, p. 78 - 84].

The drug is not effective enough in a number of categories of patients:
- not fast enough provides restoration of a reduced content of functionally full-fledged E. coli and elimination of Escherichia with hemolytic activity or poorly expressed enzymatic properties (ICS);
- does not always provide correction of immunological parameters in patients with significant disorders of the immune system, characterized by leuko-, neutro- and lymphopenia, deficiency and imbalance of individual subpopulations of T-lymphocytes, violations of the B-cell link of humoral immunity, etc.

- when used in therapeutic doses, it significantly improves, but does not completely eliminate the damaging effect on the liver tissue of oral antibiotics with hepatotoxicity;
- with simultaneous use with oral antibiotics, it reduces, but does not completely prevent the violation of indicators of homeostasis and immunobiological reactivity, for example, increased functional activity of acute phase proteins and granulocytic elastase, inhibition of phagocytic cell metabolism.

 The objective of the invention is to provide a highly effective agent for the treatment of dysbiosis, which allows to reduce treatment time, to obtain a more persistent and pronounced therapeutic effect.

The essence of the invention lies in the fact that the tool for the treatment of dysbiosis, including a lyophilized mixture of biomass of bifidobacteria and lysozyme, additionally contains sodium nucleinate in the following ratio of components per dose: biomass of bifidobacteria 10 ⁵ - 10 ⁸ CFU, lysozyme 5 - 20 mg, sodium nucleinate 50 - 250 mg.

Using the invention allows to obtain the following technical result:
- reduce by 1.5 - 2 times the treatment of dysbiosis compared with the use of bifilis in patients with severe disorders of the immune system, a significant deficiency of Escherichia coli and / or a high content of Escherichia with hemolytic and weakly expressed enzymatic properties (ICS);
- accelerate the normalization of the intestinal mucosa, its digestive, immune and barrier functions;
- reduce the risk of liver damage and provide a significant improvement in its condition under various toxic effects, especially in acute drug hepatitis;
- reduce by 2-3 times the frequency and severity of manifestations of exudative diathesis and food allergies, especially in children;
- reduce the frequency and severity of the course of infectious and destructive complications (including endogenous infection) in patients with dysbacteriosis against the background of immune disorders;
- improve tolerance and increase the effectiveness of antibiotic therapy, reduce course doses of antibiotics and the need for their prescription, for example, with selective decontamination;
- increase the effectiveness of antianemic therapy.

The technical result obtained is achieved due to the fast and stable normalization of the anaerobic and aerobic components of the intestinal microbiocenosis, protection of the liver and intestinal mucosa tissue, normalization of the cellular factors of the local gastrointestinal immunity, stimulation of the body's immunobiological reactivity (phagocyte stimulation, increased decreased lysozyme activity and antitoxic resistance), correction mechanisms of regulation of proteolysis - reduction of the initially high functional activity of the most important plasma protea noses - granulocytic elastase and correction of the activity of its main inhibitor, the protein of the acute phase of inflammation - and _1- proteinase inhibitor (a _1- PI), which takes part in controlling the mechanisms of adaptation and implementation of the inflammatory process.

For the first time, the authors established new mechanisms of action of the combination of lysozyme hydrochloride with bifidobacteria and sodium nucleinate as part of a single drug, causing the appearance of a new super-cumulative effect that exceeds the effect of separate use of individual components: correction of the functional activity of granulocytic elastase, and _1- PI and their ratios; stimulation of oxygen metabolism of phagocytic cells and serum lysozyme level; prevention of the development of acute toxic drug-induced hepatitis. In addition, the combination of sodium nucleate with bifilis or lysozyme has a more pronounced antianemic effect, obviously due to the enhancement of the indicated effect of each drug, and also due to the normalization of the content and functional activity of E. coli, which is a producer of vitamin K, a deficiency of which, especially in case of impaired function liver, leads to hemorrhagic complications and the development of iron deficiency anemia.

 It is known that bifilis as a physiologically balanced system of natural complementary gastrointestinal protection factors provides correction of intestinal microbiocenosis, increasing the content of bifidobacteria, promoting the multiplication of lactobacilli and elimination of a number of enteropathogenic and conditionally pathogenic microorganisms; improving the immunobiological reactivity of the body due to stimulation of cellular factors of local immunity, lowering the level of sensitization, increasing the level of endogenous lysozyme in the contents of the gastrointestinal tract, digestive glands and blood serum, improving the antiproteinase activity of blood plasma [patent RU N 2071339 C1, Bull. N 1, 01/10/97]. The use of biphilis and lysozyme included in its composition creates the conditions for stopping inflammatory processes, reducing the damaging effects of antibiotics and other aggressive and toxic factors on the cells of the mucous membranes, including cellular factors of local intestinal immunity, as well as on liver tissue, and has an antianemic effect blood loss, stimulates regeneration processes.

 Sodium Nucleinate is a sodium salt of low molecular weight yeast RNA. The potency of the drug includes its ability to restore the altered functions of immunocompetent and other cells of the macroorganism, enhance tissue regeneration and wound healing, increase the body's ability to neutralize toxins (tachyphylaxis), and normalize disturbed metabolic processes in the body. In this regard, the drug is used with great success as an immunomodulator for a wide range of diseases, such as pneumonia, colitis of various etiologies, ulcerative processes of the stomach, duodenum and colon, trophic ulcers, etc. Along with this, sodium nucleinate causes significant changes in microbial populations, characterized by accelerated reproduction of primarily saprophytic bacteria, representatives of normal intestinal microflora, increases the expression of adhesion molecules - key structures in the process of engraftment of normal flora in the human intestine [Zemskov V.M., Karaulov A. V., Zemskov A.M., Nazaretyan V.G. Immunomodulators in the treatment of pulmonary pathology. M., 1995, 320 pp.].

 Sodium nucleinate was not previously used to treat dysbiosis.

With the combined use of sodium nucleinate with bifilis or with lysozyme, a fundamentally new balanced protective system is formed that provides:
- faster and more persistent improvement and normalization of both the anaerobic and aerobic components of the intestinal microbiocenosis, primarily E. coli with full enzyme properties, improving their adhesion in the intestine, which leads to crowding out and elimination of E. coli with hemolyzing and reduced enzymatic properties, and also other microorganisms with pathogenic potencies;
- prevention of drug damage to ultrastructures and preservation of the physiological asynchronous mode of functioning of Paneta cells - specialized structural elements of the mucous membrane of the small intestine, producing endogenous lysozyme and IgA immunoglobulins;
- a significant reduction or leveling of the morphofunctional manifestations of acute toxic drug-induced hepatitis (normalization of the morphofunctional state of the liver tissue and the cellular composition of the liver lobules);
- pronounced stimulation of the functional activity of phagocytic cells in terms of their oxygen metabolism (spontaneous HCT test, spontaneous and induced chemiluminescence);
- correction of the functional activity of granulocytic elastase, proteins of the acute phase of inflammation (a _1- PI, and _2- macroglobulin) and lysozyme in serum;
- decrease in the degree of sensitization;
- increase of anti-infectious, antiviral, antitoxic resistance of the body;
- pronounced antianemic effect.

 The agent for treating dysbiosis is a dry microbial mass of live bifidobacteria (Bifidobacterium bifidum strain 1), lyophilized in a culture medium supplemented with lysozyme, for example lysozyme hydrochloride, sodium nucleinate and sucrose-milk medium. The drug is supposed to be released in the form of a porous or porous-crystalline mass of different shades of beige or gray, containing 5 doses of the drug in glass bottles with a capacity of 10 - 12 ml. It is possible to release the product in the form of an gro powder for use as a medicinal raw material.

One dose of the drug contains from 10 ⁵ to 10 ⁸ CFU of live bifidobacteria, 5 to 20 mg of lysozyme, for example lysozyme hydrochloride, and 50 to 250 mg of sodium nucleinate. The drug is highly soluble in water and aqueous solutions; when dissolved in water at the rate of 1 ml per dose, after 3-4 times shaking, a homogeneous yellowish cloudy suspension with a characteristic taste and smell is formed.

An agent for treating dysbiosis is made as follows. First, a uterine culture of bifidobacteria (Bifidobactetium bifidum strain 1) is grown in Blauroc medium or aminopeptide medium and then this culture is used to grow the main microbial biomass. For this, the uterine culture of bifidobacteria is seeded in a sterile reactor vessel containing cultivation medium (modified Blaurock medium or aminopeptide medium) at the rate of 6-10% of the volume of the nutrient medium. Cultivation is carried out by a deep method at 37.5 ± 5 ^o C for 16 to 24 hours, which allows to obtain biomass containing bifidobacteria 10 ⁵ -10 ⁸ CFU / ml Then the biomass is transferred into sterile containers, lysozyme, for example, lysozyme hydrochloride, sodium nucleate and a drying medium (sucrose-gelatin-milk medium) is added sequentially with constant stirring, based on the content of 0.05 - 2.0% in the final suspension of lysozyme (0, 5 - 20 mg / ml), sodium nucleinate 0.5 - 25.0% (5 - 250 mg / ml), sucrose 5 - 10% (50 - 100 mg / ml), gelatin 1 - 1.5% (10 - 15 mg / ml) and skim milk 20 - 25% (200 - 250 mg / ml). The suspension is poured into glass bottles with a capacity of 12 ml, frozen at a temperature of 40 - 45 ^o C for a period of not less than 48 hours, and then lyophilized in a vacuum dryer. Vials with the dried preparation are closed with hermetically sterile rubber stoppers and rolled around with metal caps. The whole process is carried out under aseptic conditions with phased control of the content of live bifidobacteria and the absence of extraneous microflora.

Example 1. To prepare the drug, an ampoule with a lyophilized culture of a production strain of bifidobacteria was dissolved in 5 ml of sterile distilled water and introduced into a 1-liter vial containing 100 ml of modified semi-liquid Blaurock medium. The suspension was incubated in an anaerostat at 38 ^o C for 24 hours, after which it was shaken, samples were taken to determine the microbial purity and content of live bifidobacteria. The resulting uterine culture was diluted with modified Blaurock medium to a concentration of bifidobacteria of 10 ⁶ CFU / ml, 100 ml of bacterial suspension were inoculated into a laboratory reactor, to which 1000 ml of modified aminopeptide medium was previously added and cultured for 20 h at 38 ^° C, after which they were selected samples for counting live bifidobacteria. The biomass was transferred into a separate sterile bottle and lysozyme hydrochloride, sodium nucleate and drying medium were added based on the final concentration of ingredients: 0.05% (5 mg / ml) lysozyme hydrochloride, 0.5% (50 mg / ml) sodium nucleinate, 5% (50 mg / ml) sucrose, 1% (10 mg / ml) gelatin, 20% (200 mg / ml) skim milk. After thorough mixing, the suspension was poured by an automatic dispenser into sterile F-10 glass vials placed in cassettes, which were loaded into a low-temperature chest with a temperature of about 45 ^° C for 48 hours. Then, the drug cassettes were transferred to a PG-5 vacuum dryer and lyophilized. The vials were closed with sterile rubber stoppers and circled in metal caps.

In this laboratory series of the drug, the content of lysozyme, sodium nucleinate and bifidobacteria corresponded to the calculated one and amounted to 5 mg, 50 mg, and 10 ⁵ CFU per dose, respectively.

Example 2. Obtaining a uterine culture of bifidobacteria was carried out as described in example 1. The inoculum dose of the uterine culture of bifidobacteria was 10 ⁹ CFU / ml when inoculated in a laboratory reactor with 1000 ml of amine peptide medium previously added to it. After 18 hours of cultivation at 38 ^° C, the biomass was transferred into a separate sterile bottle, after which a sample was taken to determine the microbial contamination and the content of bifidobacteria. Then, lysozyme hydrochloride, sodium nucleate and drying medium were added to the biomass with constant stirring based on the final concentration of ingredients: 20 mg / ml lysozyme hydrochloride, 250 mg / ml sodium nucleinate, 50 mg / ml sucrose, 10 mg / ml gelatin, 200 mg / ml skim milk. After thorough mixing, the suspension was poured by an automatic dispenser into 5 ml sterile vials, and further processing was carried out as described in Example 1.

In this laboratory series, the proposed drug had a single dose of bifidobacteria 10 ⁸ CFU / ml, lysozyme hydrochloride 20 mg, sodium nucleinate 250 mg.

 The new drug is intended for use in adults and children of all age groups for the treatment of intestinal dysbiosis of various origins and intestinal dysfunctions, characterized mainly by a deficiency of bifidobacteria and full-fledged E. coli, postantibiotic and toxic liver damage, iron deficiency anemia, and immune system disorders. The drug can be used against a background of antibiotic therapy.

 Mode of application. Before use, add freshly boiled and cooled to room temperature water to the bottle before use, leave for 2-3 minutes, then mix by shaking. The resulting suspension containing 5 doses of the drug should be taken 20-30 minutes before a meal. If it is necessary to use a smaller dose of the preparation (2.5 doses per dose 2 times a day), dry biomass in a vial should be previously carefully divided in half with a very clean sharp cutting object, transfer one half of the preparation to a separate clean dish and dilute in 2-3 teaspoons tablespoons of water for a single dose. The remaining part is tightly closed with a dry rubber stopper from the same bottle and left in the refrigerator until the next dose.

 When applied topically, the drug should be dissolved in the same way, moisten a sterile swab with a suspension of 2-2.5 doses per one manipulation and carefully treat the wound surface or area of the nipple and areola with this swab 20-30 minutes before the baby is applied to the breast.

 The effectiveness of the developed drug was confirmed on an experimental model of dysbiosis and acute drug hepatitis induced in rats with high doses of the tetracycline antibiotic.

 Example 3. In experiments on 30 Wistar rats with an initial body weight of 180-200 g, tetracycline was administered orally daily at a dose of 750 mg / kg on an empty stomach for 7 days. Along with the antibiotic (immediately after it), the animals were also orally administered the test drug (drug I) or bifilis (drug II) at a dose of 2.5 per rat.

 Tetracycline had a significant damaging effect on the mucous membrane of all parts of the intestine. A day after its cancellation, the animals observed the presence of dystrophic and microcirculatory changes in the mucous membrane (CO) of the duodenum, small and large intestine, which manifested itself in a violation of the structure of the villous apparatus of enterocytes, local damage to the integrity of the epithelial layer, swelling of the submucous layer of the own plate of the colon of the colon, which morphometrically noted the suppression of lymphopoiesis, changes in the number and ratios of immunocompetent cells, an increase in the number of eosinophils. Using electron microscopy, a restructuring of the cytophysiology of exocrinocytes of the intestinal glands, Panet cells, responsible for the production of endogenous lysozyme and IgA immunoglobulins, was revealed.

 The content of colibacillus significantly decreased in the contents of rat colon after administration of tetracycline, although the number of bifidobacteria, which are not known to be the predominant representatives of the normal flora of these animals, remained at the control level.

 Under the influence of a high dose of tetracycline, pronounced manifestations of its hepatotoxic effect developed, which were characterized by a morphological picture of acute toxic hepatitis: impaired microcirculation of the hepatic lobules, dystrophic changes in hepatocytes with emptying the nuclei (the “stamped core” phenomenon) and vacuolization of the cytoplasm according to the type of fat cell density, age in the periportal sections of the hepatic lobules due to an increase in the number of lymphocytes and the appearance of a significant The number of missing in normal fibroblasts, fibrocytes, eosinophilic granulocytes, which indicates intense fibrotic process with the phenomena of sensitization.

It was shown for the first time that these changes are accompanied by impaired regulation of plasma proteolysis and the functional activity of neutrophils. So, in serum, the activity of granulocytic elastase increased 1.6 times compared with the norm, which indicates a high degree of neutrophil degranulation, and the activity of a _1- PI formed in the liver increased by 2.42 times. In this case, inhibition of oxygen metabolism of peritoneal macrophages was observed (a twofold decrease in the spontaneous HCT test, spontaneous and zymosan-induced chemiluminescence), a 1.74-fold decrease in serum lysozyme level.

The use of drug I simultaneously with tetracycline exerted a pronounced protective effect, preventing the development of dystrophic and necrobiotic changes in the CO of the small and large intestines, while the villous apparatus of enterocytes normalized, and the cellular composition of the lymphoplasmacytic infiltrate of the intestine CO own plate was improved. The complete safety of ultrastructures and physiological asynchronous mode of functioning of Paneta cells was noted. Under the influence of the proposed drug, the functional activity of granulocyte elastase normalized, significantly decreasing 1.63 times compared with that when using tetracycline alone; at the same time, the activity of a _1- PI decreased 1.34 times. The positive dynamics of these indicators was accompanied by stimulation of non-specific resistance of the animal organism: the level of serum lysozyme increased on average 2.95 times compared with that with tetracycline and 1.69 times compared with the norm.

It should be noted that with the use of drug I, the indicators of nonspecific resistance did not differ much from those when used in adequate doses of lysozyme hydrochloride in combination with sodium nucleinate: the activity of a _1- PI was 9.45 ± 1.75 and 9.4 ± 1.48, respectively IE / ml, granulocytic elastase - 2.54 ± 0.31 and 2.88 ± 0.88 IE / ml and lysozyme - 1.27 ± 0.21 and 1.2 ± 0.25 μg / ml.

 The drug I almost completely prevented the development of acute drug toxic hepatitis. He ensured the normalization of key indicators of the cellular composition of the intralobular and paracentral zones of the hepatic lobules (cell density, number of endotheliocytes, lymphocytes and eosinophils); during its use, hepatocytes that had clearly contoured hyperchromic nuclei and normal glycogen content remained intact.

 Bacteriologically in the contents of the large intestine of rats, an increase of 2.5 times the titer of E. coli was detected without significant changes in the content of bifidobacteria and lactobacilli.

 The preparation II provided a slightly lower protective effect against liver tissue. Under its influence, at the end of the observation period (24 hours after the last injection of drugs), the morphometric indices of the cell composition of the lymphoplasmacytic infiltrate of the lamina propria of the small and large intestine did not fully normalize, although there was a significant improvement compared with the use of a single antibiotic. In some parts of the small intestine, microcirculatory disorders remained, while against the background of a significant improvement in the ultrastructure of the nucleus and the endoplasmic reticulum of Panet cells (compared with the use of a single antibiotic), it was approaching normal, but the ratio of cells with different electron density was not completely normalized. However, bacteriologically failed to note a significant increase in the content of Escherichia coli, which remained in the same range as when using only tetracycline.

 Under the influence of bifilisation (preparation II), a significant improvement in the state of liver tissue was noted, close to that with the use of drug I, but the latter provided a more pronounced decrease in the severity of fibrous processes in the liver tissue. So, in the periportal zones of the hepatic lobules in animals treated with bifilis or preparation I, the number of fibroblasts decreased by 2.6 and 4.2 times, respectively, of fibrocytes by 3.3 and 5.4 times compared with that when using tetracycline alone ( table 1).

In animals treated with preparation II, the activity of a _1- PI and granulocytic elastase decreased by 1.28 times and 1.33 times (p> 0.05), respectively, compared with the use of tetracycline alone, remaining above the physiological norm.

 Drug I significantly stimulated the oxygen metabolism of rat peritoneal macrophages against the background of a high dose of tetracycline, while bifilis (drug II) provided only the normalization of this integral indicator of the functional activity of phagocytes. Thus, the preparation I, when used, had a significant stimulating effect on macrophages: the spontaneous HCT test was on average twice as high as when using one antibiotic, and significantly exceeded the normal level; the indicator of stimulated chemiluminescence was higher than after a course of tetracycline alone by 4.3 times and exceeded the physiological level by 2.5 times, although spontaneous chemiluminescence reached only physiological values. After the use of preparation II, the values of the spontaneous HCT test, spontaneous and induced chemiluminescence increased, respectively, 1.56, 1.89 and 1.98 times compared with the use of tetracycline alone. Bifilisation stimulation was detected only according to the data of the spontaneous NBT test, which on average exceeded the norm by 1.76 times (p <0.05), and the induced chemiluminescence index, although normalized, was 2.19 times lower than when using the drug I (table. 2).

 Thus, an experimental study showed that the use of the proposed drug simultaneously with a broad-spectrum antibiotic tetracycline can reduce the toxic effect of the antibiotic not only on the mucous membrane of the digestive tract and cells producing endogenous lysozyme and other factors of local protection of the digestive tract, but also prevent the development of acute toxic drug hepatitis and fibrous changes in the liver tissue, to ensure the correction of the mechanisms of regulation of plasma prote olysis, stimulate factors of nonspecific anti-infectious resistance - oxygen metabolism of phagocytic cells, the level of endogenous lysozyme, prevent a sharp decrease in the content of Escherichia coli in the contents of the intestine induced by an antibiotic.

 Clinical and laboratory observations showed that the proposed tool provides effective correction of both anaerobic and aerobic components of the intestinal microbiocenosis in patients with dysbiosis, characterized by quantitative and qualitative disorders of the colic component of the intestinal microflora in combination with a deficiency of bifidobacteria and lactobacilli.

 Example 4. Patient K., 7 years old. Diagnosis: stage III dysbiosis. The girl suffers from intestinal dysfunctions (constipation, flatulence, abdominal pain, poor appetite, poor weight gain) and food allergies for 4 years. This exacerbation occurred after pneumonia and antibiotic treatment in the hospital. Biotherapy was carried out by a combination of bifilis (5 doses) with sodium nucleinate (500 mg) 2 times a day for 7 days. Clinical improvement occurred on the 3rd day of biotherapy and was characterized by normalization of the frequency and nature of the stool, a decrease in flatulence and abdominal pain, an improvement in appetite, an increase in the reduced number of red blood cells and their color index in peripheral blood. The clinical manifestations of intestinal dysfunction were completely stopped on the 5th day. Bacteriologically after the course of treatment, the content of bifidobacteria was normalized in the intestinal microflora, the total number of E. coli increased from 15 to 100 million / g, the number of coliform bacteria of CRS decreased from 95% to 65%, and the last indicator continued to decrease after the end of treatment: after 1 month. the content of E. coli CRS was 30% (Table 3). A persistent recovery was achieved when observed for 3 months.

 Thus, the use of the proposed agent had a good bacteriological effect, providing quick and lasting correction of intestinal microbiocenosis due to the restoration of the number of E. coli with full enzyme properties, elimination of Escherichia with hemolysing and reduced enzymatic properties, normalization of the number of bifidobacteria and lactobacilli.

 The proposed drug has a complex biological effect aimed at accelerating the normalization of the aerobic and anaerobic components of the microbiocenosis, mucous membrane and cellular factors of the local immunity of the digestive tract, improving the morphological and functional state of the liver tissue and increasing its resistance to drug and other toxic effects, correction of the mechanisms of regulation of proteolysis, stimulation of immunobiological reactivity and increase the adaptive capacity of the body.

Claims (1)

An agent for the treatment of dysbiosis, including a lyophilized mixture of biomass of bifidobacteria and lysozyme, characterized in that it additionally contains sodium nucleate in the following ratio of components per dose:
Bifidobacteria biomass, CFU - 10 ⁵ - 10 ⁸
Lysozyme, mg - 5 - 20
Sodium Nucleinate, mg - 50 - 250Sh

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