RU2287335C1 - Preparation "bacstitatin" for treating diseases of gastrointestinal tract - Google Patents

Abstract

FIELD: biotechnology, medicine.

SUBSTANCE: the suggested preparation "Bacstitatin" contains, weight%: sterilized culture Bacillus subtilis metabolites-containing liquid 0.1-2.0; zeolite 68-85; soybean hydrolyzate 15-30; calcium stearate 0.5-5.0. The preparation has demonstrated itself as a remedy of multiple clinical efficiency; moreover, it has been recommended in therapeutic schemes at treating patients with intestinal dysbacteriosis of different genesis (in case of chronic diseases of digestive tract, after acute intestinal infections, at the background and after the intake of antibiotics, after carrying out chemotherapy, at the background of prolonged hormonal therapy, under conditions of chronic stress states and at irrational dietotherapy). Application of the preparation considerably decreases manifestation degree of dyspeptic disorders, improves intestinal digestion, efficiently harmonizes the composition of intestinal microbiocenosis, has got immunomodeling action, positively affects psychological state in patients and favors improved quality of life in such patients.

EFFECT: higher efficiency of therapy.

3 cl, 5 dwg, 6 ex, 8 tbl

Description

The invention relates to medicine, namely to preparations for the treatment of diseases of the gastrointestinal tract (GIT) based on probiotics (living microorganisms or substances of microbial origin, which have a positive effect on the physiological, biochemical and immune responses of the host through stabilization and optimization functions of its normal microflora), and can be used in applied biotechnology, food industry, veterinary medicine and related industries.

To solve similar problems, it is currently proposed to administer orally preparations containing individual colonies of microorganisms, in particular biological preparations based on lactic acid microorganisms (EP 0097484, 1984, class A 23 C 1/05; EP 0043962, 1980, class A 23 C 19/086; US 4289888, 1979, CL A 23 C 9/123).

Preparations, as a rule, contain bacterial cells, residues of culture fluid (QOL), fats, proteins, lactose, gum and other additives. The composition of the compositions is determined by the characteristics of the microorganisms used and the nature of the task.

So, drugs based on acidophilic microorganisms are known (RU 2064269, class A 23 C 9 / 12,1994; SU 1227145, class A 23 C 9/123, 1983; RU 2080795, class A 23 C 9/12, 1997 ) containing bacterial cells and residues of culture fluid (QL) with some additives that improve its performance (gelatin, sucrose, skim milk), as well as water.

The disadvantage of such drugs is their relatively low biological activity associated with the process of deactivation of living microorganisms during their passage through the upper gastrointestinal tract (stomach, duodenum), as well as the lack of stability of their biological properties during prolonged storage, which leads to a decrease in the effectiveness of therapeutic and preventive procedures .

More stable during storage are probiotics containing spore-forming bacteria as an active principle. In particular, the probiotic Bactisubtil (Marion Merrell / France /) and its analogue Flonivin BS (Galenika / Yugoslavia /) containing the culture of the Bacillus cereus strain IP 5832 (collection of the Pasteur Institute (Paris)) were widely used. The basis of Enterohermine (firm "Sanofi Winthrop". Italy) is the culture of you.cereus. Also known is the biological product Cereobiogen ("Xing Jian", China), the active principle of which is the strain Vas.cereus DM-423 (V.V. Smirnov, S.R. Reznik, I. B. Sorokulova, V. A. Vyunitskaya. questions of the creation and use of bacterial preparations for the correction of warm-blooded microflora // Microbiol. Zhurn., 1992, v. 54, No. 6, p. 82-93).

A disadvantage of known probiotics is the limited spectrum of action, as they are directed against a narrow group of pathogens.

Known therapeutic and biological product Bactisporin for the treatment of a wide range of diseases, containing freeze-dried biomass of the strain Bacillus subtilis ZN and a protective medium based on sucrose-gelatin mixture or lactose (RU No. 2130316, A 61 K 35/66, 1999).

However, Bactisporin is not effective against a number of pathogens of acute intestinal infections, for example, caused by bacteria of the genus Pseudomonas spp., Streptococcus spp., Enterococcus spp.

Known drug Biosporin for the prevention and treatment of human gastrointestinal diseases, containing living cells of strains of Bacillus subtilis 3 (VKPM No. B-2335) and Bacillus licheniformis 31 (VKPM No. B-2336), as well as a filler (RU No. 1722502, A 61 K 35/74, 1992).

Biosporin is characterized by high antagonistic activity to a wide range of pathogenic and conditionally pathogenic microorganisms, however, the strains included in its composition are sensitive to antibiotics, therefore, the drug is not recommended for use in treatment with antibiotics.

The closest of the analogues in technical essence and the claimed effect is the probiotic "Irilis" (RU No. 2264454, A 61 K 35/66, 2005), containing live microorganisms Bacillus subtilis VKPM No. B - 8611, Bacillus licheniformis VKPM No. B - 8610, and also auxiliary additives - solvent and filler. It uses protein, organic polymer, milk, whey, albumin, sucrose, lactose, calcium stearate, glucose, sodium bicarbonate, aluminum hydroxide, methyl cellulose, talc, confectionery fat, paraffin, lanolin, cocoa butter, aluminum hydroxide gel, dextran as a filler polyglucin, starch and / or polyvinylpyrrolidone.

The disadvantage of the drug is the use of living microorganisms in its composition, which complicates or precludes its use in complex therapy with many antibiotics, as well as the presence of a negative effect on the normal microflora of the gastrointestinal tract.

The problem solved by the authors was the creation of a more effective and stable drug for the treatment of gastrointestinal diseases by introducing substances into the body that inhibit the development of pathogenic microflora and stimulate the development of beneficial microorganisms.

The technical result of the invention is to increase the effectiveness of the prevention and treatment of infectious diseases of various etiologies and dysbiosis, including in the framework of complex treatment using antibiotics by replacing live microorganisms Bacillus subtilis with their metabolites, introducing a zeolite complex with a protective medium into the composition, which can provide protection of metabolites from the action of enzymes of the gastrointestinal tract.

This problem was solved by creating a preparation that includes sterilized culture fluid (SCF) containing metabolites obtained from the cultivation and sterilization of strains of Bacillus subtilis, the introduction of which in the gastrointestinal tract inhibits pathogenic microflora, zeolite, a nutritional supplement - hydrolyzed soy flour and calcium stearate in the following ratio of ingredients (wt.%):

SCR containing metabolites of Bacillus subtilis 0.1-2.0 zeolite 68-85 soy flour hydrolyzate (fuels and lubricants) 15-30 calcium stearate (SC) 0.5-5.

The action of the drug is based on the fact that during its transit through the gastrointestinal tract in a given area, the protective capsule is destroyed and the probiotic components immobilized on zeolite particles are released into the intestinal cavity. At the same time, micellar structures are formed around zeolite particles, which are gradually released from the porous surface of the zeolite during movement along the gastrointestinal tract. On the one hand, this allows maintaining the activity of the biological components of the probiotic in the gastrointestinal tract for at least a day, which is necessary to restore and stimulate the functional activity of the normal intestinal microflora. On the other hand, the effect of the gradual release of the drug components from the zeolite surface leads to the appearance of open surfaces of its porous structure, which ensures the inclusion of ion exchange mechanisms and selective sorption of toxic compounds. This is especially important for general detoxification of the body.

The role and importance of the individual ingredients included in the drug can be determined as follows.

It is known that the metabolites of some microorganisms, in particular Escherichia coli (E. coli), obtained from cells of microorganisms placed in distilled water for 5-9 days (RU 1638156, 1991, class C 12 N 1/00), can inhibit growth pathogenic microflora and stimulate the development of normal microflora of the stomach. The authors found that the metabolites of Bacillus subtilis have a similar effect. The best results were obtained using metabolites of such strains of Bacillus subtilis as Bacillus subtilis 534 for these purposes; VKPM No. 8-2335 (3) TPI 13 - 06/07/21 - DEP / VGNKI; VKPM No. B4759 (3); p VMV105, TPI 9; rVMV 105. The obtained results are shown in table 1.

Table 1 The influence of the nature of metabolites on the development of microorganisms (zone of growth inhibition of test strains), mm: Test culture Metabolite Strain In subt
534 VKPM No. B-2335 (3) TPI 13 - 07.06.21 - DEP / VGNKI VKPM No. B-4759 (3) / rVMV105 TPI9 / rVMV105 Salmonella paratyphi 2606 18 ± 2 Salmonella stenly 5266 15 ± 2 16 ± 1 Salmonella typhimurium 12 ± 1 11 ± 1 13 ± 2 11 ± 1 13 ± 2 Staphylococcus aureus 1479 18 ± 1 19 ± 1 22 ± 3 19 ± 1 21 ± 1 Shigella sonnei 659 12 ± 2 11 ± 1 13 ± 1 11 ± 1 13 ± 1 Pseudomonas aeruginosa 32 7 ± 1 Proteid vulgaris 491 25 ± 2 Klebsiella pneumoniae 5055 16 ± 3 Citrobacter freundii 381 22 ± 2 Candida albicans 690 17 ± 1 19 ± 1 18 ± 4 19 ± 1 20 ± 2. Campilobacter jejuni 418 20 ± 4

At the same time, it was found that VKPM strains No. 8-2335 (3) TPI 13 - 06/07/21 - DEPT / VGNKI; p VMV 105 TPI 9, subt. 534, when ingested, are capable of producing 2 × 10 ⁵ ME α2-interferon. Thus, it could be expected that, when ingested, the metabolites of these strains will be able to contribute to the healing of microflora in their area of residence.

The Bacillus subtilis CGS, obtained by deep growing this microorganism, contains a unique set of biologically active components produced in the course of life. Among them, various natural antibacterial substances (bacteriocins, lysozyme, catalases) are widely represented, which selectively inhibit the growth and reproduction of pathogenic and conditionally pathogenic microorganisms in the intestine, without affecting the symbiotic microflora. In addition, microorganisms produce various enzymes and coenzymes, amino acids, polypeptides, prebiotic components that contribute to the improvement of microecological conditions in the intestine, affecting metabolic processes and having an immunomodulatory effect.

Table 2 presents the results of chromatographic analysis to determine the content of certain substances in the culture fluid after biomass removal using the example of Bacillus subtilis strain VKPM No. B-2335 (3) 3.

table 2 The composition of the sterilized culture fluid B. subtilis VKPM No. B-2335 (3) Name of the analyzed indicators SKZh Bacillus subtilis VKPM No. B-2335 (3) 3 The residual concentration (BAC) of the producer microbe, CFU / ml 0 Protein Content *, μg / cm ^-3 541.5 Proteolytic activity (PA), units PA, dm ^-3 8.7 Amylolytic activity (AA), units AA, dm ^-3 42.7 The total concentration of total amino acids, μg / cm ^-3 568.8 The total concentration of free amino acids, μg / cm ^-3 60.8 The concentration of hexosamines, mcg / cm ^-3 104.9 Glucosamine concentration, mcg / cm ^-3 95.6 The concentration of diaminopimelic acid, μg / cm ^-3 19.6 Free glucosamine concentration, mcg / cm ^-3 6.5 Adenine / Xa ** concentration, μg / cm ^-3 12.2 / 3.2 Guanine / Xg ** concentration, μg / cm ^-3 0 / 16.3 Thymine concentration / Xt **, μg / cm ^-3 8.2 / 0 The concentration of uracil / Xy **, mcg / cm ^-3 11.4 / 8.6 Cytosine / Chz ** concentration, μg / cm ^-3 0 / 12.1 Notes:
* protein content was determined by the Lowry method;
** Xa, Xg, Xm, Xy, Xc - unidentified derivatives of adenine, guanine, thymine, uracil and cytosine in terms of the basic substance.

The results of chromatographic analysis showed a significant content of nitrogen bases and their derivatives in the SCR, which possess immunomodulating properties and increase the functional activity of a single macrophage system of the body at various stages of the infectious process.

The use of SCL in concentrations lower than 0.1% leads to a significant decrease in the effectiveness of the drug (see example 3), an increase in its concentration of more than 2% is technically difficult. In addition, in this case, not all of the metabolite is sorbed on the surface of the zeolite, which does not allow to achieve significant advantages, apparently, due to the intensification of the degradation of metabolites during their transportation to the site of exposure.

As a zeolite, we used zeolite made from the natural mineral of the clinoptilolite series of the Kholinsky deposit (Buryatia), according to TU 2163-12763074-97, which is approved for use without restrictions in food and medical practice (Pylev L.N., Conclusion of the Commission on carcinogenic factors in the Ministry of Health RF, 1998). The zeolite particles are about 100 μm in size and have an oval crystal shape, which eliminates the formation of intestinal microtraumas and allows it to be used as a safe enterosorbent and a source of microelements (Avramenko V.A., Vasilevsky V.A. New sorbents based on modified zeolites and their application in ecology, agriculture and medicine // Zeolites of Primorye. Abstracts of the scientific and practical conference. Vladivostok, 1994, p.16-19; Panichev AM, Gulkov AN Natural minerals and causal medicine of the future. Vladivostok, Izdate Fate of the Far Eastern State Technical University, 2001, 216 pp .; Pylev L.N., Vasilyeva L.A., Valamina I.E. Analysis of the biological aggressiveness of zeolites of various deposits of the Russian Federation // Natural minerals in the service of man. Materials of a scientific and practical conference. Novosibirsk, 1999 p. 68-70).

Zeolite provides the transportation of metabolites in the optimal mode and the gradual release of biologically active substances immobilized on it, which allows maintaining the level of activity of the drug for at least a day. At the same time, it provides the binding and elimination of low molecular weight toxins (methane, hydrogen sulfide, ammonia, etc.), heavy metals and radionuclides. In addition, passing through the gastrointestinal tract, the zeolite is involved in selective ion exchange (removes or reduces the negative effect of aluminum ions on the body, interacts synergistically with magnesium and fluorine, and is an additional source of trace elements). Zeolite, as a source of silicon, takes part in reactions providing collagen synthesis, gives elasticity to fibrous tissues; participates in the inhibition of succinate dehydrogenase, esterase, hyaluronidase, accelerates the synthesis of irolin, glycosaminoclicans; is of particular importance for the formation of the structure of the skin, hair, nails.

The zeolite content in the preparation should ensure the sorption of the entire metabolite. A significant decrease in the concentration of zeolite leads to the loss of some metabolites and a decrease in the effectiveness of the drug. The zeolite content in a concentration of more than 85% leads to the dilution of the drug with an inactive ingredient and also reduce the effectiveness of the drug.

The soy flour hydrolyzate (GSM) in this case is, on the one hand, part of the protective environment of metabolites, which is largely responsible for the strength of their sorption on the zeolite surface, and on the other hand, is a source of amino acids that provides the nutritional needs of normal intestinal microflora and cells of the macroorganism. The main component is soy oligosaccharide (SOE), which has bifidogenic properties. It is a mixture of sucrose (44%), stachylose (23%), raffinose (7%) and monosaccharides.

Its use in a concentration of less than 15% does not provide complete protection of metabolites; its use in a concentration of more than 25% complicates the desorption of metabolites from the surface, which reduces the effectiveness of the drug and can only be recommended if a longer exposure effect is necessary with a slightly lower effectiveness.

Calcium stearate (SC) acts as a builder (aerosil). At the same time, it has an anti-stress, antioxidant effect, the presence of calcium salt improves the condition of the skeletal system, improves the activity of the nervous system.

The best results were obtained when using the composition of the following composition: sterilized culture fluid Bacillus subtilis containing metabolites, 1 wt.%. Zeolite - 78 wt.%, Soy flour hydrolyzate - 20 wt.%, Calcium stearate - 1 wt.%, Which received the code name "Bactistatin".

The drug can be produced in the form of a powder, emulsion, capsule. The latter form is the most convenient for use.

The effectiveness of the use of the drug "Bactistatin" is shown in Fig.1-5. Figure 1 shows the dynamics of changes in the composition of the microflora of the colon in patients with celiac disease before and after treatment with Bactistatin. Figure 2 shows the dynamics of changes in indicators of personal anxiety in patients with celiac disease before and after treatment with "Bactistatin". Figure 3 shows the dynamics of changes in the indicators of situational anxiety in patients with celiac disease before and after treatment with "Bactistatin". Figures 4 and 5 show the dynamics of changes in the level of depression in patients with celiac disease before (Fig. 4) and after (Fig. 5) treatment with "Bactistatin".

The drug is prepared as follows. Microorganisms Bacillus subtilis are grown by the method of deep cultivation. Then the culture fluid (QOL) with microorganisms is subjected to centrifugation and sterilization. The obtained SCL containing producer metabolites is mixed with fuel and lubricants, SC and zeolite. (Alternatively, a mixture of QOL with zeolite is sterilized.) The resulting mixture is lyophilized, in which the biologically active components of the preparation are immobilized on zeolite particles. Subsequent packaging of the composition in gelatin capsules provides protection of the components of the drug from the effects of factors causing their degradation.

Patients were prescribed the drug "Bactistatin" in a dose of 2 capsules with meals 2 times a day for at least 3 weeks. Dose selection (from 2 to 3 capsules once) and the duration of the course of treatment can be carried out individually, depending on the severity of clinical symptoms and the severity of dysbiotic disorders. Indications and contraindications for the use of the drug Supplement "Bactistatin."

Indications: 1. Dysbacteriosis of various genesis (in case of chronic diseases of the digestive tract - irritable bowel syndrome, gastritis, duodenitis, peptic ulcer, celiac enteropathy, fermentopathy, biliary tract dysfunctions); after acute intestinal infections; after taking antibiotics after chemotherapy with long-term hormonal therapy; against the background of chronic stressful conditions; against the background of irrational diet therapy.

2. Immunodeficiency conditions.

3. Prevention of bisbiotic disorders in infectious and viral diseases.

4. Non-alcoholic and alcoholic steatohepatosis, steatohepatitis.

5. Allergic diseases (popinosis, vasomotor rhinitis, chronic urticaria, Quincke's edema).

6. Dermatological diseases (neurodermatitis, eczema, psoriasis).

7. Prevention of complications after surgical interventions in abdominal surgery, traumatology, gynecology.

When prescribing antibiotics, Bactistatin is used from the first day of antibiotic therapy and at least two weeks after its completion in order to prevent the development of antibiotic-associated diarrhea and intestinal dysbiosis.

When staying in conditions of environmental and professional stress (adverse environmental and climatic conditions), Bactistatin is prescribed throughout the entire adaptation period.

Bactistatin is fully compatible with all components of eradication anti-Helicobacter pylori therapy and contributes to its effectiveness and safety.

There are no contraindications to the use of the drug, however, "Bactistatin" is not prescribed for individual intolerance to the components.

Example 1. Obtaining the claimed drug

A probiotic preparation is prepared as follows. Bacillus subtilis microorganisms are grown by deep cultivation in a fermenter having a volume of 130 l. After the cultivation process, centrifugation is carried out. The culture fluid (QL) is mixed with zeolite, previously ground to a particle size of 100 μm, and sterilized by autoclaving for 30 min at 80 ° C. Into the obtained sterile culture fluid (SCL), predetermined amounts of a soy flour hydrolyzate and calcium stearate are introduced. The resulting complex is lyophilized. The yield of the obtained product is 200 ± 5 kg. If necessary, the resulting product is Packed in gelatin capsules of 0.5 or 1 g.

Example 2. The study of the effects of the composition of the claimed drug on the inhibition of the growth of pathogenic and conditionally pathogenic microflora.

Table 3 shows the effects of a dose of 500 mg of the drug, introduced in MPA, on the inhibition of growth of Sh.sonnei and S.aureus 1479.

Table 3 Inhibition of the growth of microorganisms on MPA containing 500 mg of probiotic The composition of the drug, wt.% Zina of growth inhibition of Sh.sonnei after 24 hours, mm Zone of growth inhibition of S.aureus 1479 after 24 hours, mm SKZH zeolite Fuels and lubricants SC one 78 twenty one 16.7 24.2 0.1 85 fifteen 5 13.5 15.3 2 78 19.5 0.5 16,44 22.5 one 68 thirty one 15,2 16,2

The presented results show that when 500 mg of the drug is introduced into MPA in the following ratios of components: SKZh-1%, zeolite-78%, GSM-20% and SK-1%, maximum inhibition of Sh.sonnei growth (16.7 mm) is ensured S.aureus 1479 (24.2 mm). In this case, a more effective exit of antibacterial metabolites from the drug occurs, which contributes to the selective inhibition of the growth of pathogenic and conditionally pathogenic microorganisms. Further studies were conducted on the preparation of the above formulation, which received the code name "Bactistatin".

Example 3. The study of the effects of various doses of the drug "Bactistatin" on the growth of normal microflora.

The effect of the drug on the growth of normal microflora was studied on the example of Escherichia coli M-17 microorganisms. The daily culture of E. coli M-17 in a volume of 0.1 ml was introduced into tubes containing 10 ml of meat-peptone broth (MPB). Test tubes with BCH contained the following concentrations of the claimed drug - 0.1 mg / ml; 1.0 mg / ml; 500 mg / ml and 1000 mg / ml. A daily culture of E. coli M-17 was also added to the control series of tubes with BCH, but there was no probiotic concentration in these media. Crops were cultivated at + 37.0 ° С for 24 hours. After cultivation, 0.1 ml of MPB were sown on meat peptone agar (MPA), on which after 24 hours the grown colonies of microorganisms were counted and their concentrations calculated (CFU / ml). Microorganism concentrations were expressed as the natural logarithm (Ln C, CFU / ml). The results of the study are presented in tables 4 and 5.

Table 4 Concentrations of Escherichia coli M-17 (CFU / ml) in a liquid nutrient medium containing various doses of the probiotic "Bactistatin" The cultivation time of E. coli, M-17, hours The dose of the probiotic "Bactistatin", mg / ml Concentration of E. coli M-17, Ln C, CFU / ml, M ± m, δ 0 0 4.91 ± 0.07 (δ = 0.16) 24 0 10.3 ± 0.13 (δ = 0.33) 0 0.1 4.91 ± 0.07 (δ = 0.16) 24 0.1 11.09 ± 0.07 (δ = 0.19) 0 1,0 4.91 ± 0.07 (δ-0.16) 24 1,0 11.42 ± 0.08 (δ = 0.21) 0 500,0 4.91 ± 0.07 (δ ⁼ 0.16) 24 500,0 13.70 ± 0.24 (δ = 0.91) 0 1000,0 4.91 ± 0.07 (δ = 0.16) 24 1000,0 10.65 ± 10.14 (δ = 0.35) Note:
- Ln C is the natural logarithm of the concentration of E.coli M-17;
- M ± m is the average value and the error of the natural logarithm of the concentration of E. coli. M-17;
- δ is the standard deviation.

Table 5 The significance of differences between the concentrations of Escherichia coli M-17 obtained by culturing microorganisms on the control (*) and experimental nutrient media (**) containing various doses of the probiotic "Bactistatin" The cultivation time of E. coli M-17, hours The dose of probiotic in a liquid nutrient medium, mg / ml Concentrations of E. coli, M-17 in a liquid nutrient medium, Ln C, M ± m, CFU / ml Fisher criteria (F) and significance level (P) The effectiveness of increasing concentrations of E. coli M-17 in experimental nutrient media relative to the control nutrient medium, (%) 24 0,0 10.3 ± 0.13 * - - 24 0.1 11.09 ± 0.07 ** F = 2.39
P = 0.15 (> 0.05) 7.67 24 1,0 11.42 ± 0.08 ** F = 1.96
P = 0.21 (> 0.05) 10.87 24 500 13.70 + 0.24 ** F = 0.123
P = 0.01 (<0.05) 33.01 24 1000 10.65 ± 0.14 ** F = 0.87
P = 0.43 (> 0.05) 3.39

где: С_о - опытные значения натурального логарифма концентрации E.coli M-17, полученные при культивировании микроорганизмов на жидкой питательной среде, содержащей различные дозы пробиотика «Бактистатин»; С_к - контрольные значения натурального логарифма концентрации микроорганизмов, полученные при культивировании микроорганизмов на жидкой питательной среде, не содержащей пробиотик «Бактистатин»; статистически значимые различия между опытными и контрольными значениями концентраций Е.coli M-17 подчеркнуты.Note: the effectiveness of the increase (%) in the concentration of E. coli M-17 in experimental nutrient media relative to the control nutrient medium was determined by the formula (1)

where: С _о - experimental values of the natural logarithm of the concentration of E.coli M-17 obtained by culturing microorganisms in a liquid nutrient medium containing various doses of the probiotic "Bactistatin"; With _to - control values of the natural logarithm of the concentration of microorganisms obtained during the cultivation of microorganisms in a liquid nutrient medium that does not contain the probiotic "Bactistatin"; statistically significant differences between the experimental and control concentrations of E. coli M-17 are underlined.

The presented results indicate that when a dose of 500 mg / ml is added to the BCH, there is a significant stimulation of the growth of E. coli M-17. Compared with the control values of the concentrations of microorganisms, the effect of increasing their number is 30%. This suggests that for effective stimulation and restoration of normal microflora of the gastrointestinal tract, the optimal dose of the claimed probiotic composition is in the range of 400-600 mg.

Example 4. Evaluation of the effectiveness of the use of biologically active additives "Bactistatin" in patients with erosive antral gastritis after a course of eradication therapy.

An open study of the effectiveness of the use of the drug “Bactistatin” included 30 patients with H. pylori-associated acid-dependent diseases of the stomach and duodenum. The history of peptic ulcer disease was 6 ± 2 years. In all patients, peptic ulcer was in the acute stage. The age of patients ranged from 20 to 50 years, the average age was 42 ± 4 years. Patients were divided into two groups of 20 people - the test group (patients received the drug at a dose of 2 capsules 2 times a day with Bactistatin food in the treatment regimen) and 10 - the control group (patients received only basic therapy).

The research results are presented in table 6. As can be seen from the table, "Bactistatin" is effective for stopping the phenomena of gastric and intestinal dyspepsia in patients with acid-dependent diseases of the upper gastrointestinal tract. The use of this drug in patients with erosive antral gastritis led to a clear positive clinical effect in most patients, manifested in the form of a decrease in symptoms of intestinal dyspepsia, developing against the background of eradication therapy.

During the treatment with Bactistatin, all patients noted good tolerability of the drug, no side effects were detected. Some of the patients after the course of treatment continue to take these drugs, noting a significant improvement in subjective sensations.

Table 6 The dynamics of the severity of clinical manifestations in patients with erosive antral gastritis in the main (test) and control groups before and during treatment. Symptom Patients of the test group Patients in the control group before treatment (%) 10 day of treatment (%) 30 day of treatment (%) before treatment (%) 10 day of treatment (%) 30 day of treatment (%) Stomach ache 90 60 0 90 60 0 Heaviness and discomfort in the abdomen. 90 ** fifty 10** 90 ** 60 40 ** Nausea 40 10 10 70 twenty 10 Heartburn 60 ** 0 ** 10 90 fifty twenty Vomiting twenty 0 0 10 0 0 Burping 90 * 40 * twenty 40 40 40 Dry mouth 40 thirty twenty 0 0 0 Bitterness in the mouth 60 * twenty* 0 twenty twenty 0 Rumbling 90 fifty* twenty* 90 90 70 Bloating 90 ** 10** 0 twenty 90 fifty Decreased appetite fifty* 10* twenty 45 40 25 Frequency of stool (once a day) 0.86 * 1.06 1.13 0.86 2.06 1.7 ** Bristol chair character 2.56 * 3 * 3.67 * 3,5 5.2 ** 4.01 Notes: * p <0.05 (differences are not statistically significant)
** p <0.01 (differences are statistically significant)

Example 5. Evaluation of the effectiveness of the use of "Bactistatin" for the treatment of patients with celiac disease with dysbiotic disorders of the colon of varying severity.

Celiac disease is a chronic progressive disease characterized by diffuse atrophy of the mucous membrane of the small intestine, which develops as a result of intolerance to the protein of cereal gluten. The severity of the disease is evaluated depending on the severity of malabsorption syndrome and the duration of the disease and the degree of change in intestinal microbiocenosis.

Under supervision were 7 people with celiac disease. All patients had a clinical diagnosis of celiac enteropathy previously based on the clinical course of the disease, morphometric examination of the duodenal mucosa, and immunological blood tests (determination of the level of antigliadin antibodies and antibodies to transglutaminase). The drug "Bactistatin" was prescribed for four weeks, 2 capsules 2 times a day. Analysis of feces for dysbiosis was performed before treatment and on 25-31 days after treatment. The results of the study of changes in microbiocenosis in patients with celiac disease before and after treatment are presented in table 7 and figure 1 (the concentration of the gastrointestinal microflora is presented as Lg CFU / g feces).

The study of microbiocenosis in patients with celiac disease revealed significant changes in the composition of the colon microflora, characterized by a significant decrease in indicators. In 71.4% of patients, the number of bifidoflora was reduced, in 28.6% of patients the number of bifidobacteria was at the level of 10 ⁴ -10 ⁵ cells / g (with a norm of 10 ⁸ -10 ¹⁰ cells / g). In 14.3% of patients, before the treatment of bifidobacteria, the feces were not determined. Lactobacilli were below normal in 100% of patients. In 42.9% of patients, there was a decrease in bacteroids in feces, in 14.3% of patients, bacteroids were not determined. Marked changes were observed in the qualitative and quantitative composition of E. coli: in 42.9% of patients, E. coli with normal enzymatic properties was not detected, in 42.8% of patients its quantity was reduced, only in 14.3% of patients the number of E. coli was sufficient (10 cells / g). 42.9% of the total number of E. coli was esirichia with altered enzymatic properties (normally no more than 10%). In 14.3% of patients, feces of the genus Candida were found in feces. An increase in the number of clostridia was noted in 14.3% of patients, the number of which reached 10 ⁸ .

Table 7 Changes in the composition of the colon microflora after treatment with Bactistatin in patients with celiac disease. The name of the group of microorganisms the number of patients with a modified indicator before treatment indicator change after treatment (number of patients (%) -Percentage is indicated on the number of changed indicators) increase in quantity quantity reduction did not change Bifidumbacteria 5 (71.4%) 4 (57.1%) 1 (14.3%) 0 Lactobacilli 7 (100%) 4 (57.1%) 1 (14.3%) 2 (28.6%) Bacteroids 3 (42.9%) 3 (42.9%) 0 0 E. coli with normal enzymatic activity 6 (85.7%) 6 (85.7%) 0 0 E. coli with reduced enzymatic activity 4 (57.1%) 1 (14.3%) 3 (42.9%) 0 Enterococci 5 (71.4%) 0 1 (14.3%) 3 (42.9%) Hemolytic microorganisms 1 (14.3%) 0 1 (14.3%) 0 Opportunistic bacteria 3 (42.9%) 1 (14.3%) 1 (14.3%) 1 (14.3%) Staph. aureus 0 0 0 0 Staph. saprophyticus, epidermidis 0 0 0 0 Candida mushrooms 1 (14.3%) 0 1 (14.3%) 0 Clostridium 1 (14.3%) 0 1 (14.3%) 0

On the background of taking the drug "Bactistatin", a significant improvement was noted in indicators of both the anaerobic flora and the aerobic component. An increase in bifidobacteria and lactobacilli was noted in 57.1% of patients, bacteroids increased in 42.9%; E. coli indicators improved - an increase in the number of Escherichia with normal enzymatic activity was observed in 85.7% of patients. In patients, the percentage of E. coli with altered properties decreased from 42.9% to 28.6%. After the therapy, the number of hemolytic organisms, opportunistic bacteria, yeast-like Candida and clostridia decreased by 14.3%.

Example 6. Assessment of changes in the dynamics of the psycho-emotional sphere in patients with celiac disease before and after the use of "Bactistatin"

In this group of patients with a diagnosis of celiac disease (7 people), an assessment of changes in psycho-emotional status was performed before and after the use of Bactistatin. In this case, the levels of anxiety and depression were determined.

Before treatment, an increase in the levels of personal (ITT (ST-L)) and situational (ITT (ST-S)) anxiety was revealed in 71.4% and 28.6% of patients, respectively. Against the background of Bactistatin therapy, the levels of general personal and situational anxiety, emotional discomfort (ED), phobic component (FOB), social protection (SZ), asthenic component (ACT) and perspective assessment (OP) decreased. The dynamics data of indicators of personal and situational anxiety and their components are presented in table 8 and figure 2 (dynamics of indicators of personal anxiety in patients with celiac disease before and after treatment) and figure 3 (dynamics of indicators of situational anxiety in patients with celiac disease before and after treatment).

Table 8 Dynamics of the level of increased personal and situational anxiety before and after treatment with Bactistatin Personal anxiety: before treatment,% of patients after treatment,% of patients Total 71,4 42.9 Emotional discomfort 85.7 71,4 Asthenic component 85.7 57.1 Phobic component 57.1 42.9 Perspective assessment 71,4 57.1 Social protection 42.9 28.6 Situational Anxiety: Total 28.6 14.3 Emotional discomfort twenty 0 Asthenic component 60 54 Phobic component 34 twenty Perspective assessment 40 34 Social protection twenty 7

Data on the dynamics of the level of depression (UD) before and after treatment are presented in figures 4 and 5.

The level of depression (UDS) in solo celiac disease before treatment with Bactistatin was increased in 71.4%, with a moderate degree of depression was detected in 28.6%, mild - in 42.8%. After the treatment, a moderate degree of depression was observed in 14.3%, mild - in 14.3%. This means that against the background of treatment with Bactistatin, positive changes are noted in the form of a decrease in the levels of anxiety and depression. In addition, in patients receiving Bactistatin, based on the data of the SF-36 questionnaire, a significant improvement in the quality of life was recorded on almost all scales characterizing their physical, social and psychological state, which indicates an increase in their quality of life.

The absence of side effects of "Bactistatin" allows you to extrapolate the possibility of its appointment to all patients with intestinal dysbiosis, as well as healthy individuals with a preventive purpose.

Thus, “Bactistatin” proved to be a tool with multifaceted clinical efficacy; it is currently recommended in treatment regimens for the treatment of patients with intestinal dysbiosis of various genesis (for chronic diseases of the digestive tract, after acute intestinal infections, during and after taking antibiotics , after chemotherapy, against the background of prolonged hormonal therapy, in conditions of chronic stressful conditions, with irrational diet therapy).

The use of the drug significantly reduces the severity of dyspeptic disorders, improves intestinal digestion, effectively harmonizes the composition of the intestinal microbiocenosis, has an immunomodulating effect, positively affects the psychological status of patients and helps to improve their quality of life.

"Bactistatin" has no contraindications and does not cause side effects: "Bactistatin" is not prescribed for individual intolerance to the components. In most cases, other therapeutic and wellness products (antibiotics, vitamins, enzymes, microelements, etc.) are not needed when using the drug, as they are already contained in them and (or) are replaced by similar ones in their effect.

The data presented indicate that the new probiotic (synbiotic) "Bactistatin" has advantages over the known probiotics, synbiotic and prebiotics in the ability to correct intestinal microbiocenosis disorders. "Bactistatin", harmonizing the state of intestinal microbiocenosis, improves psychological status, has an immunomodulating effect, positively affects the clinical picture of the disease and provides a significant improvement in the quality of life of patients

The main advantages and distinguishing features of "Bactistatin" in comparison with other means of correcting dysbiosis in general can be represented as follows:

- contributes to the improvement of microecological parameters in the intestine through various mechanisms, thereby creating conditions for the growth of symbiotic microflora;

- has a pronounced selective inhibitory effect in relation to the conditionally pathogenic and pathogenic flora:

- has a pronounced immunomodulatory effect;

- normalizes digestion processes;

- has an antitoxic effect;

- maintains its activity at an optimal level during the day, due to the gradual release of biologically active components;

- does not contain live microbes;

- the shelf life of the drug is 2 years.

Claims (4)

1. A drug for the treatment of diseases of the gastrointestinal tract, characterized in that it contains a sterilized culture fluid containing metabolites of Bacillus subtilis, a nutritional supplement - soy flour hydrolyzate, zeolite and calcium stearate in the following ratio of ingredients, wt.%: Sterilized culture fluid containing metabolites of Bacillus subtilis 0.1-2.0 Zeolite 68-85 Hydrolyzed Soybean Meal 15-30 Calcium stearate 0.5-5.0 2. The drug according to claim 1, characterized in that it contains a sterilized culture fluid containing metabolites of Bacillus subtilis - 1.0 wt.%, Zeolite - 78 wt.%, Soy flour hydrolyzate - 20 wt.%, Calcium stearate - 1 wt.%. 3. The drug according to claim 1, characterized in that it is a powder. 4. The drug according to claim 1, characterized in that it is enclosed in an enteric-soluble capsule coated with an acid-protective layer.

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