Biological preparation balis for preventing and treating infectious diseases

FIELD: medicine.

SUBSTANCE: invention refers to biotechnology, namely to biological preparations. The biological preparation for preventing and treating infectious diseases and dysbioses of various aetiology containing a living microbial mass of Bacillus subtilis VKPM B-8611 and Bacillus licheniformis VKPM B-8610 strains, filtered living cultures of Bacillus subtilis VKPM B-8611 and Bacillus licheniformis VKPM B-8610 strains and a lectin-binding substance of a culture fluid supernatant of the living microbial mass of Lactobacillus fermentum 90 TS-4(21) strain of molecular weight min. 20 kDa with certain relation of the ingredients.

EFFECT: biological product higher clinical effectiveness in infectious diseases and dysbioses of various aetiology, tuberculosis infection, including XDR TV caused by polyresistant tuberculosis mycobacteria.

3 cl, 1 dwg, 7 tbl, 4 ex

 

The invention relates to biotechnology and for the creation of new probiotic preparations on the basis of bacteria of the genus Bacillus, which can be used for the prevention and treatment of infectious diseases of humans and dysbiosis of various etiologies.

It is known that a major problem in the treatment of tuberculosis is multiresistance of bacteria to antibiotics, that creates an insurmountable obstacle to reducing the incidence of TB extensively drug-resistant (so-called XDR-TB or XDR-TB). When XDR-TB, in addition to drug resistance, characteristic of multiresistentsuse adds resistance to all fluoroquinolones and at least one of three injectable second-line drugs (capreomycin, kanamycin or amikacin) (who Report no WHO/HTM/TB/2010.3). Currently, there are strains of Mycobacterium tuberculosis resistant to three or four or five or even six antibiotics, including antibiotics reserve, and therefore the question of search of drugs, allowing to solve the problem.

Described treatment-and-prophylactic probiotic agent (EN 2314819 C1, Lalic and others, 20.01.2008)biomass bacteria spores of the genus Bacillus. Contains the bacterial strain Bacillus subtilis VKPM B-7048, and/or the bacterial strain Bacillus subtilis VKPM B-7092, and/or bacterial strain Bacillus licheniformis VKPM B-7038. Ensure the leads to a significant expansion of the range and increase the biological activity of therapeutic-prophylactic probiotic means however, the possibility of recovery of sensitivity to antibiotics under the action of the tool.

Well-known drug (RU 2403260 C2, SPODSBJERG and others, 10.11.2010, WO 2006/097110 from 21.09.2006 contents and objectives), the producer of which is a Bacillus licheniformis, which according to the chemical structure relates to the peptides. This drug has a broad antimicrobial activity against infectious pathogens of both prokaryotes and eukaryotes. However, it is reported that treatment of drug-resistant strains of Mycobacterium tuberculosis in a joint application with antibiotics.

The present invention is a drug for the treatment of infectious diseases and especially chronic tuberculosis XDR TV, caused by Mycobacterium extensively drug-resistant.

The biological product for the prevention and treatment of infectious diseases, mainly tuberculosis and associated candidiasis and dysbiosis of various etiologies, including live microbial mass from strains of Bacillus subtilis VKPM B-8611 and Bacillus licheniformis VKPM B-8610, characterized in that it further comprises a filtrate of live cultures mentioned strains of Bacillus subtilis and Bacillus licheniformis and legislatively substance from the supernatant culture fluid of live microbial mass strain Lactobacillus fermentum 90 TS-4(21) in the following components:

live microbial mass strain of Bacillus subtilis VKPM B-8611 - not less than 5×108CFU;

live microbial mass strain of Bacillus licheniformis VKPM B-8610 - not less than 1×107CFU;

the filtrate live microbial mass strain of Bacillus subtilis VKPM B-8611, dried and 2.5-4.5 mg;

the filtrate live microbial mass strain of Bacillus licheniformis VKPM B-8610, dried and 2.5-4.5 mg;

legislatively substance from the supernatant culture fluid of live microbial mass strain Lactobacillus fermentum 90 TS-4(21) with a molecular mass of more than 20 kDa, dried - 5,5-7,5 mg

Infectious diseases include chronic tuberculosis, intestinal infections, including hemorrhagic colitis caused by E. coli O-157, nosocomial surgical infections, Candida, dysbiosis. Chronic tuberculosis may be caused by mycobacteria XDR TV with extensively drug-resistant.

The technical result of the invention is to increase the effectiveness of antibiotic treatment of TB, including XDR TV, due to multidrug-resistant Mycobacterium tuberculosis.

Included in the biological strains of Bacillus subtilis VKPM B-8611 and Bacillus licheniformis VKPM B-8610 deposited in the Federal GOSNIIGENETIKA (FGUPGosNIIgenetika), the date of the international Deposit 12.08.2004). Live cultures of both strains are characterized by a wide spectrum of antagonistic activity, high proteolytic activity, with what osobnosti to the production of bacteriocins, enzymes lysozyme, amylase, peptides. Bacterial filtrates of live cultures of both strains containing the above extracellular products of their activity, showed unexpected activity against pathogenic mycobacteria of the tuberculosis disease. In addition to the antagonistic action - the ability to retard the growth of bacteria, changing the sensitivity of pathogens to antibiotics, increasing activity spectrum antibiotic, restoring sensitivity to 1, 2, 3, 4 or 5 antibiotics. It should be noted that a common problem for TB patients receiving prolonged antibiotic therapy is the development of dysbiosis and candidiasis, as an extreme degree of dysbiosis. This drug is a multicomponent probiotic. It contains added legislatively complex, which blocks the adhesion of Candida to epithelial cells of the terminal ecological niches of the microorganism, thereby prevents the development of candidiasis. To prepare legislative substance, part of the probiotic drug used producer probiotic drug Lactobacterin. He was selected variant Lactobacillus fermentum 90-TS-4 (21) (clone 3), agglutinates in the presence of concanavalin A. immunology and have high ability to Express legislative component in the environment is Multivitamine. This glycoprotein complex prevents the colonization of yeast-like fungi of the genus Candida on the epithelium (see EN 2367686 C1, Anokhin and others, 20.09.2009). Under the action of the probiotic composition Balys about 80% of drug-resistant pathogenic mycobacteria restore its sensitivity to antibiotics, strains of XDR TV increased sensitivity to 1-5 antibiotics.

Strains of B.subtilis 07 (VKPM B-8611) and B.licheniformis 09 (VKPM B-8610) isolated from healthy wheat plants deposited in Russian national collection of industrial microorganisms and are characterized by the following properties.

Bacillus subtilis VKPM B-8611 - gram-positive aerobic spore-forming Bacillus size of 2.7 to 0.6×0,8-0,7 μm, arranged singly or in the form of chains. Cell mobile form in aerobic conditions the spores oval, which are located in the cell Central. When sporoobrazovanie cells do not swell.

On the environment Gause No. 2, wort-agar, environment Gromyko strain grows abundantly, forms a Mat folded colony of flesh-colored with rugged edges, easy to remove the loop from the agar.

In the cytoplasm of strain after growth on glucose agar not detected include poly-β-hydroxybutyric acid. On BCH culture forms a film.

Does not grow in anaerobic conditions, not hydrolyzes urea, does not form gas from nitrate under anaerobic conditions is s, does not produce argininosuccinate. Culture forms a catalase. Gives a positive reaction Voges-Proskauer, grows in the presence of 7% NaCl. Hydrolyzes starch and casein, thins gelatine. Ferments glucose, arabinose, xylose, mannitol with the formation of acid without gas. Reduces nitrates, discolor methylene blue. Has no coagulates and lecithinase activity, has high proteolytic and amylase activity.

Bacillus licheniformis VKPM B-8610 - gram-positive spore-forming bacilli, the amount of 2,6-0,7 x 0,5-06 μm. Cells are motile, peritricha, primarily in the form of chains. Spores oval, located in the Central cell. Cells when sporoobrazovanie not inflated. After growth on glucose agar in the protoplasm are not detected include poly-β-hydroxybutyric acid.

On the IPA forms colonies with a dull rough surface, opaque, tightly attached to the agar often on the surface is detected mucus. On the BCH is formed wrinkled film, sometimes with cream shade.

Culture produces catalase, characterized by the ability to grow on agar under anaerobic conditions. Gives a positive reaction Voges-Proskauer, grows at 7% NaCl. Hydrolyzes starch, casein, not hydrolyzes urea. Gelatin liquefies slowly. Ferments glucose, arabinose, xylose, man is it with the formation of acid without gas. Reduces nitrates in the anaerobic conditions of the nitrate forms a gas. Produces argininosuccinate, lysozyme. Has no coagulates and lecithinase activity.

The basic properties of the strains Century licheniformis VKPM B-8610 and B. subtilis VKPM B-8611 presented in table 1.

Strains of B.subtilis VKPM B-8611 and B.licheniformis VKPM B-8610 characterized by high antagonistic activity against wide spectrum of pathogenic and conditionally pathogenic microorganisms and resistance to several antibiotics (Table 2, 3). These properties allow you to apply Balys simultaneously with antibiotics, with the strengthening of the common antibacterial effect.

As part of the proposed biological Balys these strains can be used in various combinations, for example in equal proportions (titer cells): biomass of strain .subtilis VKPM B-8611 in titer of 1·109CFU/ml and biomass of strain B.licheniformis in titer of 1·109CFU/ml or in any ratio in the range of (1-100):(100-1), for example, the biomass of the strain .subtilis VKPM B-8611 in the titer of 5 x 109CFU/ml and biomass of strain B.licheniformis VKPM B-8610 in titer of 1·109CFU/ml, or biomass of strain .subtilis in titer of 1·1010CFU/ml and biomass of strain B.licheniformis PMBC No-8610 in title 5·109CFU/ml, or biomass of strain .subtilis VKPM B-8611 in titer of 1·109CFU/ml and biomass of strain B.licheniformis VKPM B-8610 in titer of 1·107CFU/ml, etc.

To obtain sterile filtrate the living cultures of the strains subtilis VKPM B-8611 and B.licheniformis VKPM B-8610 cultivated separately at constant shuttlebay at 37°C. Biomass is filtered through sterilizing filters with a preliminary centrifugation (g=9859,6 m/s2). The pH was adjusted to 6.0±0,05.

The target product is extracellular products of vital activity of strains of B. subtilis VKPM B-8611 and B.licheniformis VKPM B-8610. The filtrates live cultures subjected to freeze-drying and the resulting substance stored at a temperature not above 20°C. the Substance is stable, does not change in the process of freeze-drying and during storage for 2 years.

In the manufacture of the drug used dried substance, but it is possible and the use of the effluent in liquid form. As part of the proposed drug Balys this substance can be used in an amount of 3 mg/ml

To obtain legislatively substance as a producer used Lactobacillus fermentum strain 90-NS-4(21) (strain deposited in Russian national collection of industrial microorganisms under registration number In-7573). After sieving strain L. fermentum 90-TS-4(21) were sampled colonies on the basis of agglutination with Kona on the glass at a concentration of at least 1.75·10-3mg/ml

Selected variant strain L. fermentum 90-TS-4 (21) were cultured on liquid medium MPC-1 in a 1000 ml in CO2-conditions for 15-17 hours at a temperature of 37(±0,05)°C and constant stirring.

After 15 to 17 hours of culture cells using zentrifugenbau the Oia (14-16 min, 3000 rpm (g=9859,6 m/s2)) was separated from the culture fluid.

Supernatant culture fluid was transferred to the cell company AMICON (USA) (2000 ml) and was freed from low molecular weight components and the residual amount of bacterial cells using sterilizing membranes brand "Mile-long" with a pore diameter of 0.02 mm.

Purified culture fluid was concentrated at the cell company AMICON (USA) (2000 ml) using membranes "Diablo" stamps UM-20 to volume of 100 ml at the same time the supernatant culture fluid washed three times with distilled water from lactic acid, bringing the pH to a value of 6.0±0,05.

The resulting concentrate of the culture fluid of strain L. fermentum 90 TS-4 (21) were incubated for 24 hours at room temperature with Kona-separate. After days adosados was removed, and con-sepharose washed three times with phosphate buffer (pH of 8.2). Next was added saline solution (pH 3.0) and incubated for 3 hours at room temperature. Collected adosados and brought it up to a pH of 7.2 with 0.1m NaOH. The presence legislatively substance controlled using a 0.1% solution of Koh in the reaction of calcarenitic.

The target product is anti-adhesive component on the basis of legislatively structures strain L. fermentum 90 TS-4(21) has the following characteristics:

the component containing the 150,0 mg/ml protein, aggregative newsto is constant at pH values above 6.0;

the molecular mass determined using polyacrylamide gel electrophoresis, is from 25 to 30 kDa;

properties legislatively substance is stable and does not change during storage and during lyophilization.

Legislatively substance has a modulating effect on the adhesive activity of the test cultures of yeast fungi of the species C. albicans and conditionally-pathogenic strains of the species E. coli (see EN 2367686).

Modulating the property fraction, devoid of the ability to respond with Kona, and faction, capable of reacting with Kona concentrate cultural liquid (QL) L.fermentum strain 90 TS-4 (21) in the test of inhibition of adhesion of test-cultures of epithelial cells of the vagina (EV), showed the following (see Table 4). Chromatographically purified fraction legislatively active substance inhibits the adhesion of vaginal isolate of yeast fungi of the species C. albicans strain 04.703 B in epithelial cells and to a lesser extent affects the adhesion of oral isolates of yeast fungi of the species C. albicans. Adhesion of E. coli strain K 12 fraction has no effect, but substantially blocks the adhesion of E.coli strain 89-1449 epithelial cells of the vagina. At the same time when removing legislative component is enhanced adhesive activity part of the test cultures of E. coli to epithelial cells of the vagina. Adhesion vaginal isolate of drozhzhepodobny the fungi species .albicans strain 04.703 in this model system was significantly decreased. These data confirm that you have received legislative component with a strong modulatory activity against high adhesive strains of yeast-like fungi species .albicans and other opportunistic pathogens.

When composing multicomponent probiotic preparation Balys ratio of live strains of VKPM B-8611 and .licheniformis VKPM B-8610 can be changed at specific intervals, as described above. The filtrates of live cultures of strains VKPM B-8611 and .licheniformis VKPM B-8610 (extracellular substance) were added in amounts of about 3 mg Legislatively substance Lactobacillus fermentum 90 TS-4 (21) was added in an amount of about 6 mg biologic Balys also contains a protective environment for preservation of bacteria during processing, the final prescription form and subsequent storage. As a protective environment it may contain, for example, Saharsa gelatin Wednesday, milk powder, gelatos, lactose, sucrose, etc. biological Balys may further comprise a solvent. The solvent may be used, for example, distilled or boiled water, saline, etc.

The biological Balys can optionally also contain a filler, usually used in the manufacture of different prescription forms. When formvariables it may contain, for example, dextrans, polyglucin, starch, polyvinyl pyrrolidone, sucrose, lactose, calcium stearate, glucose, sodium bicarbonate, aluminium hydroxide, methylcellulose, talc, etc. While getting a suppository or candles as filler it contains, for example, confectionery fat, paraffin, lanolin, cocoa butter, gel, aluminum hydroxide, etc.

The biological product may be encapsulated or immobilization on different types of media or sorbents, such as aerosol, cellulose, charcoal, carboxymethyl cellulose, hydroxyethyl cellulose, chitosan, etc.

The biological product may also be dried.

The biological Balys can be used for oral, or vaginal, rectal, or topical use in the form of an aqueous suspension. The mechanism of action of biological Balys-based adhesive and antagonistic activity of bacteria-probiotics. This effect is due to the production of various physiologically active substances, displacing pathogenic and conditionally pathogenic microorganisms from the digestive tract and has a stimulating effect on specific and nonspecific defenses of the host.

The invention is illustrated by the following examples.

Example 1. Obtaining liquid drug

The strains were cultured separately or napotnik or liquid nutrient media.

When stationary technology cultivation of strains spend on solid agar media under mattresses or in glass bottles thermostatted shaker at a temperature of from 22 to 38°C for 12-16 h to 7 days. After incubation wash the biomass grown on the surface of the medium, stabilizer - protective medium containing 5%solution of lactose, are brought together in a ratio of 10:1, is poured into vials. Receive a biological product with a biomass content of strain B. subtilis VKPM B-8611 in titer of 1·1010CFU/ml and biomass of strain Century licheniformis VKPM B-8610 - 1·109CFU/ml To the obtained bacterial composition add 2.5-4.5 mg/ml of the filtrate indicated strains and 5.5-7.5 mg legislatively substance Lactobacillus fermentum 90 TS-4 (21).

When industrial technology cultivation of strains is carried out in the reactor/fermenter with a nutrient medium for culturing at a temperature of 35-38°C for from 10 to 18 o'clock Process is considered complete, if the cell concentration is 4-5 billion/ml and the ratio of spores and vegetative cells of 1:1. After incubation separately grown cultures are brought together in a ratio of 1:1 and add saharso gelatin protective environment, poured into vials. Receive a biological product with a biomass content of strain .subtilis VKPM B-8611 in the titer of 5 x 109CFU/ml and biomass of strain .licheniformis VKPM B-8610 - 5·109The OE/ml To the obtained bacterial composition add 2.5-4.5 mg/ml of the filtrate indicated strains and 5.5-7.5 mg legislatively substance Lactobacillus fermentum 90 TS-4 (21).

Example 2. Receiving the drug in the form of a lyophilisate

When stationary technology cultivation of strains spend on solid agar media under mattresses or in glass bottles thermostatted shaker at a temperature of from 22 to 38°C for 12-16 h to 7 days. After incubation wash the biomass grown on the surface of the medium, a protective medium containing 10%glycerin solution to the bacterial composition add 2.5-4.5 mg/ml of the filtrate indicated strains and 5.5-7.5 mg legislatively substance Lactobacillus fermentum 90 TS-4 (21).

Poured into ampoules (vials) or in cassettes stainless steel, and then subjected to freezing and dehydration by vacuum-drying plant or spray dried method.

When industrial technology cultivation of strains is carried out in the reactor/fermenter with a nutrient medium for culturing at a temperature of 35-38°C for from 10 to 18 o'clock Process is considered complete, if the cell concentration is 4-5 billion/ml and the ratio of spores and vegetative cells of 1:1. After incubation separately grown cultures are brought together in a 2:1 ratio and add stabilizer is PR, to the obtained bacterial composition add 2.5-4.5 mg/ml of the filtrate indicated strains and 5.5-7.5 mg legislatively substance Lactobacillus fermentum 90 TS-4 (21).

Poured into ampoules (vials) or in cassettes stainless steel, and then subjected to freezing and dehydration by vacuum-drying plant or spray dried method.

Example 3. Receiving the drug in pill form

Cultures of strains .subtilis VKPM B-8611 and .licheniformis VKPM B-8610 after adding the components of the suspension, to the obtained bacterial composition add 2.5-4.5 mg/ml of the filtrate indicated strains and 5.5-7.5 mg legislatively substance Lactobacillus fermentum 90 TS-4 (21), dehydrated by vacuum drying or spray drying plant, combined with sugar granules, sliding substances (starch or calcium stearate and compressed on a rotary press.

Example 4. Receive preparation in the form of suppositories

Cultures of strains .subtilis VKPM B-8611 and .licheniformis VKPM B-8610 after adding the components of the suspension, to the obtained bacterial composition add 2.5-4.5 mg/ml of the filtrate indicated strains and 5.5-7.5 mg legislatively substance Lactobacillus fermentum 90 TS-4 (21); dehydrated by vacuum drying or spray drying plant, combined with fillers (konditerski fat, paraffin and other) and cast on a special installation suppositories.

Example 5. All obtained in examples 1 or 2, or 3, or 4 ways and forms of biological Balys check emissions on laboratory animals, specific antagonistic activity against the test cultures - representatives of different groups of pathogenic and conditionally pathogenic microorganisms and antibiotic resistance.

The biological Balys characterized by harmlessness. The definition of safety was performed on 2 groups of mice: 1-St group used for the study of drug Balys. In the 2nd group were investigated on the safety of the filtrates of live cultures. For each variant of experiment and control used at least 10 mice weighing 15-16, To determine the safety of the drug Balys the contents of the vial were dissolved in 0.5 ml of saline and was administered this dose orally to mice 1-th group. The drug is considered harmless if all mice remain alive for 5 days of observation, and none of them identified the disease. The 2 groups of mice were injected intraperitoneal injection of sterile filtrates from drug Balys, weekly for 4 weeks in a volume of 0.3 ml At the beginning and end of the experience from the tail vein of mice took a drop of blood for research leukogram by counting the cells in smears with staining by Romanovsky - the Institute at the end of the experiment was carried out weighing mice and compared with the initial weight, also conducted post-mortem examination of animals. Mouse 1-St group during 5 days of observation were alive, none of them identified diseases. Mouse 2-nd group within 30 days live, and gained weight 2.0 to 2.5, At postmortem examination revealed a slight increase in spleen, and other organs without visible changes. Morphological study of white blood cells gave the following results: in mice groups 1 and 2 (blood taken on the last day of experience) indicators leukogram did not differ from the original.

The biological Balys characterized by a broad spectrum of antagonistic activity against the test strains cultures of pathogenic and conditionally pathogenic microorganisms. The study carried out by the method of deferred antagonism. For this purpose the contents of the vial are dissolved in 1 ml of physiological solution. Received a suspension detect touch on diameter Petri dishes with agar medium Gause No. 2. Sowing incubated in a thermostat at 37°C for 72 h and Then grown culture podshuveyt stroke test microorganisms (500 millionth suspension subsistence crops in saline solution). Analysis performed after 18 hours incubation at 37°C largest zones of no growth of the test cultures.

Control of the growth of the test cultures is parallel virusiv is of them on plates with agar medium Gause No. 2 without the studied culture.

The optimal number of living cells in a single dose is 1-5×109. Further increase in the number of microbial cells does not change significantly antagonistic activity of the drug against the test cultures of microorganisms (see Table 3).

Legislatively substance has a modulating effect on the adhesive activity of the test cultures of yeast fungi species .albicans and conditionally-pathogenic strains of the species E. coli to epithelial cells of the vagina of healthy women. This statement is illustrated by the following results (see Table 4).

Spore probiotics have a set of properties that allows you to compete with pathogenic and conditionally pathogenic microorganisms. These properties include: antagonistic activity, the capacity for adhesion to epithelial cells, a certain level of resistance to hydrochloric acid, bile, etc. But still not studied the action of these drugs on the causative agents of tuberculosis infection. In recent years, along with increased morbidity and mortality from tuberculosis has increased the number of multi-drug resistant TB (MDR-TB) and strains with extensively drug-resistant (XDR-TB) Mycobacterium tuberculosis. The clinical picture of drug-resistant tuberculosis manifests itself in the patient when the population is Oia multidrug-resistant Mycobacterium far exceeds the population of bacilli, sensitive to drugs. When studying the antagonistic activity of the drug Balys used reference strains Academia (M.tuberculosis), Vallee (M. bovis), Ravenal (M. bovis), GIS (M.avium)obtained from gisk named after. Laurasia. As indicator strains were also used isolated from cattle and we identified cultural, biochemical and chemotaxonomic methods strains .bovis (No. 3, Ryazan) - see Table 5, 6. In addition, 20 patients with tuberculosis were isolated and identified 20 drug-resistant strains of M. tuberculosis (Table 7). Method to study the antagonistic activity of probiotic strains against Mycobacterium tuberculosis involved the use of culture filtrates of liquids, obtained through sterilizing filters (see Lazovsky A.L., and others, the Effect of probiotics on pathogenic mycobacteria // Problems of tuberculosis and lung diseases. Publishing house "Medicine", M., 2006. No. 7. - 25-27). Analysis was performed within 25-30 days incubation at 37°C index block growth (IBR) of mycobacteria. IBR was determined by calculating the ratio of the number of colonies grown on the control medium without probiotic, to the number of colonies grown on the medium with a probiotic. The maximum IBR considered the number 10.

Effect spore probiotics on medicinal Chu is stateliest mycobacteria studied on 20 clinical strains isolated from TB patients people. Used the above processing methods spore probiotics. The drug sensitivity was determined in accordance with Order No. 109 from 21.09.03. "On improving TB control in the Russian Federation". The results are presented in the figure. In this experiment, we used the following drugs in different concentrations: 1, 2 - streptomycin, 10 and 25 µg/ml, respectively; 3, 4 - isoniazid 1 and 10 μg/ml; 5 - kanamycin 30 µg/ml; 6 - ethambutol 2 µg/ml; 7 - rifampicin 40 μg/ml; 8 - ethionamide 30 µg/ml; 9 - ofloxacin 2 µg/ml; 10 - capreomycin, 30 μg/ml; 11 - PASK (paraaminosalicilovaya acid) 1 mg/ml

All reference strains studied probiotics showed antagonistic activity. As shown in Table 5-7, spore probiotics inhibited the growth. M.tuberculosis. Index block growth (IBR) in 20 strains of Mycobacterium tuberculosis ranged from 1 to 10.

The study of drug susceptibility of isolates grown on media with a sterile filtrate showed that restoration of drug sensitivity was 80%, and 95% of the original cultures were resistant to 1, 2, 3, 4, 5, 6 or even 7 anti-TB drugs. 20% of strains were resistant to 7-8 antibiotics, i.e. treat XDR-TB and MDR-TB. 20 drug-resistant clinical strain is in mycobacteria in 16 (80%) recovered sensitivity to one, two, three, four or five drugs.

1. The biological product for the prevention and treatment of infectious disease and dysbiosis of various etiologies, including live microbial mass from strains of Bacillus subtilis VKPM B-8611 and Bacillus licheniformis VKPM B-8610, characterized in that it further comprises a filtrate of live cultures mentioned strains of Bacillus subtilis VKPM B-8611 and Bacillus licheniformis VKPM B-8610 and legislatively substance from the supernatant culture fluid of live microbial mass strain Lactobacillus fermentum 90 TS-4(21) with a molecular mass of more than 20 kDa in the following components of 1 unit of a biological product:
live microbial mass from a strain of Bacillus subtilis VKPM B-8611 - not less than 5×108CFU;
live microbial mass from a strain of Bacillus licheniformis VKPM B-8610 - not less than 2×108CFU;
the filtrate live microbial mass strain of Bacillus subtilis VKPM B-8611, dried and 2.5-4.5 mg;
the filtrate live microbial mass strain of Bacillus licheniformis VKPM B-8610, dried and 2.5-4.5 mg;
legislatively substance from the supernatant culture fluid of live microbial mass strain Lactobacillus fermentum 90 TS-4(21), lyophilized - 5,5-7, mg.

2. Biological preparation according to claim 1, intended for the treatment of infectious diseases: chronic tuberculosis, intestinal infections, nosocomial surgical infection, candidiasis and dysbiosis.

3. Biological preparation according to claim 2, intended for the treatment of chronic tuberculosis, caused by Mycobacterium XDR TV with extensively drug-resistant.



 

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3 cl, 6 tbl, 5 ex

FIELD: medicine.

SUBSTANCE: invention concerns medicine, namely to phthisiology and can be used for preventing and treating tuberculosis. A method involves the introduction of 150-200 ml of herbal tea containing root of inula, ginseng, eleutherococcus, peony, valerian, golden root, green oats, haws, juniper, leaves of motherwort and foxglove in the following proportions respectively 3:0.02:0.02:0.03:2:0.02:12:3:3:1:1 in the morning on an empty stomach. Then in the afternoon 10-30 minutes before meals 150-200 ml of herbal tea containing herb of Icelandic moss, great nettle, pimpernel, wormwood, bearberry, bryony, bottle brush, hops and root of inula in the following proportions respectively 2:3:1:1:2:1:2:1:2 is taken. In the evening 10-30 minutes before meals 150-200 ml of herbal tea containing herb of sage, bur marigold, origanum, thyme, leaves of coltsfoot, fruits of fennel and root of inula, hairgrass, marshmallow and parsley, corn silk in the following proportions respectively 1:2:1:1:3:1:3:2:3:2:3 is prescribed. The therapeutic course makes 21 days.

EFFECT: using the invention provides more effective treatment and prevention of tuberculosis.

4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to new substituted quinoline derivatives of general formula

, including any their stereochemical isomer forms, and to their pharmaceutically acceptable salts wherein p means an integer equal to 1; q means an integer equal to 1 or 2; R1 means halogen; R2 means C1-6alkyloxy; R3 means aryl; each R4 and R5 independently means C1-6alkyl; or R4 and R5 together with nitrogen atom whereto attached, form a radical selected from the group consisting of piperidino and piperazino with each radical to be substituted by 1 substitute selected from C1-6alkyl; R6 means aryl1; R10 means C1-6alkyl or aryl C1-6alkyl; Z means S or NR10; aryl and aryl1 represent phenyl. Also, the invention refers to a pharmaceutical composition based on the compound of formula (la), to the use of the compound of formula (la) and to a method for preparing it.

EFFECT: there are prepared new quinoline derivatives of formula (la), effective in treating a bacterial infection.

15 cl, 4 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to phthisiology, pulmonology, thoracic surgery. Method includes standard anti-tuberculosis treatment. Into pleural cavity, without extraction of pleural fluid introduced is mixture of medications, consisting of 10 thousand units of contrical, 5 thousand units of heparin and 30 mg of prednisolone at the background of standard antibacterial treatment. Introduction of medication mixture is carried out on 5-6 ml of pleural fluid. If treatment of pleurisy is started in due time, mixture is introduced single time, in case of total, chronic or recurring pleurisy mixture is introduced 2-3 times with 7-10 day intervals.

EFFECT: method ensures reduction of disease treatment terms, due to efficient elimination of inflammation and prevention of immunodeficient state caused by loss of protein substances, contained in pleural fluid, is economical and non-burdensome either for patient or for physician.

2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to phthisiology and pharmaceutical industry. A method for preparing a water-soluble chitosan para-aminosalicylate consists in preparing an aqueous solution of para-aminosalicylic acid (2.8 wt %) at temperature 90°C, adding low-molecular chitosan (low molecular weight 3.9 and 30 kDa) in amount 2.8 wt % in mixing with a mechanical mixer, dispersing a reaction mixture at temperature 85°C for 40 minutes. After termination of the process, the reaction mixture is vacuumised; a solid residue is dried in vacuum at temperature 30°C.

EFFECT: preparation of medicines for tuberculosis.

3 ex, 9 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention relates to novel antibacterial chinolin derivatives of formula (1a), their stereochemically isomeric forms, N-oxides and pharmaceutically acceptable salts and solvates , where p equals 1; q equals 0, 1, 2, 3 or 4; R1 represents hydrogen, halogen, aryl or Het; R2 represents hydrogen or alkyloxy; R3 represents arylalkyl; each R4 and R5 independently represent hydrogen or alkyl; R7 represents hydrogen, alkyl or aryl; where aryl is selected from phenyl or naphtyl and is optionally substituted with 1, 2 or 3 substituents selected from hydroxy, halogen, cyano, nitro, amino, mono- or dialkylamino, alkyl, C2-6alkenyl, optionally substituted with phenyl, halogenalkyl, alkyloxy, halogenalkyloxy, carboxyl, alkyloxycarbonyl, aminocarbonyl, morpholinyl or mono- or dialkylaminocarbonyl; where Het is selected from furanyl, thienyl, pyridinyl, benzofuranyl, optionally substituted with 1, 2 or 3 substituents selected from halogen, hydroxyl, alkyl or alkyloxy.

EFFECT: compounds can be used for creation of preparations on their base for treatment of bacterial infection

21 cl, 4 ex, 3 tbl

FIELD: medicine.

SUBSTANCE: invention relates to veterinary science. An active agent for preparing a veterinary drug for treating or preventing Clostridium perfringens in poultry is presented by 3-0-acetyl-4"-0-isovaleryl-tylosin or its pharmaceutically acceptable acid addition salt.

EFFECT: preparing the veterinary drug for treating or preventing Clostridium perfringens in poultry.

10 tbl, 6 ex

FIELD: medicine.

SUBSTANCE: invention refers to method, and concerns a method for making a lyophilised antiviral agent consisting of dissolving a peptide in water, mixing the prepared solution with a number of structure-forming substances, sterilising and lyophilising including freezing out and drying. As a peptide, the peptide His-Gly-Val-Ser-Gly-Trp-Gly-Gln-His-Gly-Thr-His-Gly is used.

EFFECT: invention provides optimising the process enabling preparing the broad-spectrum lyophilised antiviral agent corresponding to the certified pharmacopeial description of the manufacturer.

3 cl, 1 ex, 3 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to peptide-based compounds containing three-member rings containing a heteroatom, which efficiently and selectively inhibit specific activity of N-terminal nucleophilic (Ntn) hydrolase, bonded with a proteasome. The peptide-based compounds contain epoxide and are functionalised at the N-end.

EFFECT: peptide-based compounds exhibit anti-inflammatory properties and cell proliferation inhibition, oral administration of said peptide-based proteasome inhibitors is possible owing to bioavailability thereof.

23 cl, 14 ex

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of general formula , where R1 denotes OH, OPO3H2 or OCOR5; R2 denotes H, OH or OPO3H2; A denotes N or CR6; R3 denotes fluorine; R4 denotes H, C1-3alkyl or C3-6cycloalkyl; R5 denotes an alanine residue; R6 denotes H, C1-6alkoxy group or halogen; and n=0 or 1; and to pharmaceutically acceptable salts of compounds of formula I. The invention also relates to a pharmaceutical composition having antibacterial activity, and to use of compounds of formula I to obtain a medicinal agent for preventing or treating bacterial infections.

EFFECT: compounds of formula I, having antibacterial activity.

14 cl, 3 dwg, 2 tbl, 14 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely dermatovenerology and is applicable for treating chronic infectious urethritis complicated by prostatitis. That is ensured by taking azithromycin 1.0 g (sumamed) once a day in 1st-7th-14th days combined with the complex therapy. The complex therapy involves rectal suppositories Dalargex, 1 suppository daily at bedtime No.10, prostate massage, rectal laser therapy and daily application of fluoroquinolone - gemifloxacin (factive) 1 tab. (320 mg) a day for 14 days.

EFFECT: method enables prolonged remission of the disease, higher life quality of the patients, recovers the reproductive function due to improved eradication of the opportunistic flora in the prostate secretion contents.

4 ex

FIELD: medicine.

SUBSTANCE: group of inventions refers to medicine and concerns recombinant viral vaccines. Substance of the inventions involves recombinant viral vaccines containing a recombinant poxvirus vector which expresses in vivo at least one heterological sequence of a nucleotide which codes an antigen and one 1H-imidazo [4,5-c] quinoline-4-amine derivative with said poxvirus vector being MVA. Particularly, the presented invention discloses the combined vaccines which combine the recombinant viral vectors MVA containing a nucleotide coding one or more HPV antigens which are able to intensify the immune response which is observed in in vivo under said recombinant viral vectors.

EFFECT: advantage of the group of inventions consists in intensifying the immune responses.

18 cl, 2 ex, 10 dwg

FIELD: medicine.

SUBSTANCE: invention relates to veterinary science. The preparation contains pumpkin sludge 84-86%, walnut concentrate 10-12%, the preparation Metrosept 2-3%, emulsifier T2 1-2%.

EFFECT: preparation is safe for animals, effective in treating inflammatory processes and wound surfaces, possesses prolonged action.

4 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, obstetrics, gynecology and includes estimation of puerperas' state, depending on which treatment tactics is selected. Severity of state is estimated by two groups of criteria of table No 3, presented in the description, by 1 point: absence of factors of risk of purulent-septic complications (PSC), subfebrile condition with single rise to 38°C, arrested by antibacterial therapy (ABT), absence of easily arrested intestine paresis after Cesarean section, the cervix of uterus is formed, presence of uterus involution in hysteroscopy at the background of treatment with endometrectomy or vacuum-aspiration, USE-data - increase and extension of uterus cavity by 0.5-1.0 cm, absence of deformation in the area of scar or deformation up to 0.5 cm, on uterus walls - linear echo-positive structures up to 0.2-0.3 cm thick, local sections of reduced myometrium echogenicity in the region of scars not larger than 1.5x1.5 cm, absence of infiltration, hematoma in the region of scars, improvement of laboratory indices in dynamics. Criteria of group 2 are estimated in 2 points: presence of PSC risk factors, long-lasting fever with resumption after finishing ABT, intestine paresis with absence of effect from intensive or repeated treatment courses, absence of tendency to formation of uterus cervix, stable uterus subinvolution, by USE data: extension of uterus cavity ≥1 cm, deformation in scar ≥0.5 cm, echopositive structures ≥0.4 cm, reduces echogenicity in scar zone ≥2.5×1.5 cm, hematoma or infiltrate in retrovesical space, in area of scars, absence of positive dynamics or change of laboratory indices to the worse. If state of puerpera is characterised by criteria of the first group to 7 points, it is estimated as uncomplicated form of disease, if - more than 7 points or at least by one criterion from group 2, as complicated form.

EFFECT: method ensures reliable justification of adequate tactics of patient management, strict control of their state in dynamics, reduction of frequency of complicated generalised forms of PSC, terms of staying in hospital.

2 ex, 3 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to novel substituted pyrimidine derivatives, having HIV replication inhibiting properties, or pharmaceutically acceptable salts thereof. In formula (1): R1 denotes hydrogen; R2 and R3 independently denote hydrogen; R7 and R8 denote C1-6alkyl; R4 denotes cyano; R9 denotes C1-6alkyl optionally substituted with cyano, C2-6alkenyl substituted with cyano, C2-6alkynyl optionally substituted with cyano; R5 denotes C1-6alkyl optionally substituted with Ar or Het; C2-6alkenyl optionally substituted with Ar or Het; C2-6alkynyl optionally substituted with Ar or Het; C3-7cycloalkyl; Ar; Het; R6 denotes H, Het; Y denotes -OR11, -NR12R13; R11 denotes hydrogen or C1-6alkyl optionally substituted with hydroxy, C1-6alkoxy or pyridyl; R12 denotes hydrogen or C1-6alkyl; R13 denotes hydrogen or C1-6alkyl; or R12 and R13 together with a nitrogen atom, which is substituted by said two substitutes, form a morpholinyl; imidazolyl; X denotes -NR1-; Het denotes 5- or 6-member completely saturated ring, where one or two ring members are heteroatoms, each independently selected from nitrogen and sulphur, and where the rest of the ring members are carbon atoms; and where any member of the heterocycle with a nitrogen heteroatom can optionally be substituted with C1-6alkyl; where the 5- or 6-member ring can optionally be annelated with a benzene or thiophene ring; each aryl independently denotes phenyl or phenyl substituted with one substitute selected from C1-6alkoxy.

EFFECT: high efficiency of using said compounds.

7 cl, 4 ex, 1 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to novel azoles of general formula 1A and 1B and pharmaceutically acceptable salts thereof, having activity on hepatitis C and hepatitis GBV-C virus. Said compounds have NS5A viral protein ligand properties and can be used as active components for a pharmaceutical composition and a medicinal agent for treating diseases caused by said viruses. In general formula 1A and 1B, the solid lines accompanied by dotted lines denote a single or double bond, wherein if one of them is a single bond, the other is a double bond; X and Y optionally assume different values and denote a nitrogen, oxygen or sulphur atom or a NH group; R1 and R2 optionally denote identical radicals 2.1-2.20, in which the asterisk (*) indicates site of the bond to azole fragments. Said fragments and values of A and B are given in the claim.

EFFECT: more value of the compounds.

10 cl, 1 tbl, 16 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to neurology, and can be used in treatment of muscle spasticity in patients with spinal cord injury. For this purpose botulotoxin is introduced into iliolumbar muscle under control of X-ray computer tomography. Introduction of preparation is performed by dorsal access into proximal part of iliolumbar muscle at the level of LII-LIII vertebrae, 2-3 cm more laterally than spinous process. Depth of introduction is 8-10 cm.

EFFECT: method makes it possible to ensure efficient relief of local spasticity of iliolumbar muscle with absence of botulotoxin effect on other groups of muscles due to accurate introduction of preparation into definite region of said muscle.

2 ex

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