Complex preparation-probiotic in immobilized and lyophilized form and method for its obtaining

FIELD: medicine, veterinary science, agriculture, biotechnology.

SUBSTANCE: the present innovation refers to preparations of medicinal and veterinary indication and methods for their obtaining. The suggested complex preparation-probiotic includes a carrier being a porous aluminum oxide-based enterosorbent SUMS-1 and cells of bacteria-eubiotics with nutritive and protective media immobilized upon the mentioned carrier-enterosorbent. The cells of bacteria-eubiotics with nutritive medium are being a lyophilized concentrate of the consortium of either bifidobacteria or lactobacteria, or their mixture at titer being 108-1010 CFU/g, and as a porous carbon-modified enterosorbent SUMS-1 it contains enterosorbent SUMS-1 additionally treated with the solution of metal salt at the following ratio of the preparation components in lyophilized form, weight%: cells with nutritive medium of the consortium of bifido-, or lactobacteria-eubiotics, or their mixture with titer of 108-1010 CFU/g 1.0-10.0; protective medium 0.1-10.0; enterosorbent SUMS-1 additionally treated with the solution of metal salt - the rest up to 100%. Method for obtaining the suggested preparation deals with treating a porous enterosorbent SUMS-1 with the solution of metal salt at volumetric ratio being 1:1, not more and its washing against a reagent with 2-3-fold volumes of distilled water followed by mixing enetrosorbent and bacteria-eubiotics with nutritive and protective media at the ratio of 1:1, not more to keep the mixture till complete immobilization of bacterial cells onto enterosorbent with its subsequent freezing and freeze drying before sterilization. As bacteria one should apply the concentrate of the consortium of bifidobacteria, or lactobacteria, or their mixture at titer being 108-1010 CFU/g. The suggested complex preparation-probiotic provides higher efficiency of sorption and desorption of bacterial cells with enterosorbent SUMS-1 that shows its clinical efficiency and higher colonization activity of bacteria-eubiotics.

EFFECT: higher efficiency.

9 cl, 5 ex, 2 tbl

 

The invention relates to preparations for medical and veterinary use and methods for their preparation and can be used in biotechnology, medicine and agriculture.

For the prevention and treatment of many pathological conditions caused by disorders of endoecology, widespread chelators. These are different types of active carbons, for example, karbacher, carblog, microsorb, polifan etc. (Sanov P.L. Pharmaceutical chemistry. - M.: Medicine, 1978, p.132-133; the patent of the Russian Federation N2029546, IPC6AC 9/16, publ. 27.02.95,), natural materials and their analogues (crystal microcellulose, zeolites, chitosans, pectins etc), synthetic mineral sorbents and other materials that are used to cleanse the body of toxins, radionuclides, heavy metals and toxins. Chelators reduce the effects of Exo - and endotoxemia.

However, chelators do not eliminate the causes of the effects of Exo - and endotoxemia, as a rule, they are not selective, working primarily on an excess of substrate, can have a negative effect due to sorption necessary for the body metabolites (vitamins and hormones) and do not provide active normalization of microflora.

The function of the normal microflora in the body are vital and very extensive: synthesizing, regulatory, defensibility the existing, detoxification, enzyme, protective and immunostimulirutuyu. For normalization of microflora of widespread drugs of live bacterial cells of bifidobacteria (int. application (WO) No. 85/03848), MKI AS 9/12, publ. 12.09.85 g), lactic acid bacteria (EPO application No. 0154614, MKI AS 9/123, publ. 11.09.85,), colibacteria etc.

The disadvantages of these drugs are relatively short shelf life if they are in liquid form, or high cost, if they are made in the dry form, as well as poor resistance to inactivating factors of the external environment and the gastrointestinal tract of the body when applied.

Known integrated product that includes a porous inorganic carrier and immobilized therein cells of microorganisms (application Germany No. 3410650, MKI 12N 11/14, publ. 03.10.85 year). As carriers used stalactite substrate with a dual pore structure. Substrates are through macropores that allow for the free exchange of liquid and gas from the inner part of the carrier in the environment. The size of the macropores is in the range of the values of the cells of microorganisms.

The disadvantage of this drug is that it is not intended for use as a medical or veterinary drug in the absence of data on the impact of stalactite media caused by nanam microorganisms on the microbiocenosis of the gastrointestinal tract of the human or animal.

Known complex pharmaceutical preparation comprising a pharmaceutically acceptable carrier and immobilized microorganisms on it (int. application (WO) No. 91/09608, MKI AC 35/74, publ. 11.07.91 year). The media does not impair the viability of the microorganisms. As microorganisms use the time duration L1A strain of Streptococcus lactis. The product is designed to combat pathogenic intestinal microflora of humans and animals, causing diarrhea and other gastrointestinal disorders.

The disadvantage of the drug is that it uses the time duration L1A strain of Streptococcus lactis it mainly affects pathogenic intestinal flora of humans and animals, i.e. exhibits antagonistic properties, but little is involved in the restoration of the microflora of the gastrointestinal tract. In addition, there are no data about antacid properties of the medium, which indicates a weak protection carrier immobilized cells of Streptococcus lactis under the action of the internal environment of an organism with low pH values.

Known complex bacterial preparation comprising a carrier and immobilized thereon microbial cells at a concentration of 1 mm22,1×103of-1.0×105(patent RF №2017486, IPC6AC 31/00, publ. 15.08.94 year). As the carrier used activated charcoal in powder form with a particle size less than 30 microns, and as microorganisms COI is lsout Lactobacillus and Escherichia coli.

The drawback of this drug is that the activated carbon used as the carrier, is temporily sorbent with a predominance of micropores and a high specific surface. Consequently, the microbial cells are located on the surface of the particles of sorbent and inadequately protected under adverse environmental effects. Activated carbons are actively absorb low molecular weight biologically active substances, vitamins and intestinal gases that degrade the performance of the peristalsis. They quickly become clogged and work mostly in the upper regions of the gastrointestinal tract. In addition, the coals do not have antacid properties, which is not conducive to the survival of covered cells under the action of environments with low pH values. The drug does not involve the use of multiple strains of organisms of different species, for example a mixture of bifidobacteria and lactobacilli, which greatly reduces the effectiveness of the product especially when restoring microflora of the gastrointestinal tract.

Known probiotic drug in immobilized form, including the media, representing the sorbent, and cells eubiotics, immobilized on the specified carrier, and the cells eubiotics use in conjunction with a nutrient medium and in use as a sorbent is a material with antacid properties, developed meso - and macroporous structure and volume of macropores is not less than 0.01 cm3/g in powder form with a particle size of not more than 0.1 mm, or in the form of granules of size (0.1 to 5.0 mm), or in the form of tablets with the following proportions of the components of the drug, wt. %: cells eubiotics with a nutrient medium and a titer of 108-108CFU/ml to 1.0-50,0; media-sorbent - the rest is up to 100%.

As the sorbent using a porous material based on aluminum oxide with hydrophilic-hydrophobic toponimia surface, such as enterosorbent type SMS-1 modified with carbon, or enterosorbent type SIAL, modified silicone polymer. As eubiotics use bifidobacteria or lactobacilli or their mixture (patent RF №2118535, IPC6AC 35/74, publ. 10.09.98,).

However, used in the specified drug sorbents have insufficient efficiency of sorption and desorption of bacteria-eubiotics, and has a low colonization activity included preparation of bacteria-eubiotics.

A method of obtaining complex preparation of bacteria-eubiotics (RF patent No. 2067114, IPC AC 35/74, publ. 27.09.96 year), which includes the cultivation in submerged cultivation of culture strains of bifidobacteria, and Streptococcus, mixing the biomass with a protective medium of the following composition (wt.%): sakh the rose 25-30; gelatin 2-3; milk powder 3-5; MSG 0.5 to 1 and contact-sorption dehydration of the target product, they have to be cooled to minus 8-10°With a sorbent, such as alumina, with a residual moisture of less than 1% in mass ratio of 1:10-12, respectively, and the final drying is carried out for 18-20 hours at a temperature of 0-5°in a confined space.

However, used in a specified way sorbent has insufficient efficiency of sorption and desorption of bacteria-eubiotics, and has a low colonization activity of the bacteria-eubiotics.

The closest technical solution (prototype) is a probiotic drug in immobilized and dried form (RF patent No. 2164801, IPC AC 35/74, publ. 10.04.01). The preparation includes the media, representing the sorbent, and cells of probiotics, immobilized on the specified carrier, and the cells of probiotics used in conjunction with a nutrient medium and in use as a sorbent material with antacid properties, advanced meso - and macroporous structure and volume of macropores is not less than 0.01 cm3/g in powder form with a particle size of not more than 0.1 mm, or in the form of granules of size (0.1 to 5.0 mm), or in the form of tablets according to the invention the preparation additionally contains a protective environment, and the cells of probiotics with a nutrient medium, not only is jut a dry concentrate of bifidobacteria in milk-based with title 10 8-1010CFU/g and amino acid content (mg/100 g dry biomass): glutamic acid - 15,0-68,0; glutamine - 8,0-20,0; leucine - 10,0-50,0; arginine - 20,0-30,0; cysteine - 50,0-80,0 in the following ratio of components of the drug in dry form wt.%: cells eubiotics with a nutrient medium and a titer of 108-1010CFU/g - 0,1-10,0; protective environment - 0,1-10,0; media-sorbent - the rest is up to 100%. As the protective environment of the use of gelatin, sucrose, ascorbic acid and sodium chloride at the following content, wt.%: gelatin - 8,5-11,5; ascorbic acid - 0,8-1,2; sodium chloride - 1,7-3,0; sucrose - the rest is up to 100%. As the sorbent using a porous material based on aluminum oxide with hydrophilic-hydrophobic toponimia surface, such as enterosorbent type SUMS-1-modified carbon. The drug as a bacteria-probiotics further comprises dry yeast viscous strains of acidophilic bacteria Lactobacillus acidophilus type AB, the ratio of which with a dry concentrate of bifidobacteria is 1:(8-9), respectively. As of bifidobacteria in the dry concentrate is used Bifidum bacteria bifidum No. 1 or No. 791, or two-3 or Bifidobacterium longum 379, or Bifidobacterium adolescentis MC-42, or a mixture of bacterial cells of the indicated strains, taken in equal parts by weight.

The closest way (prototype) is a way of receiving the Oia complex preparation of probiotic in immobilized and dried form, contained in the description to the patent of Russian Federation №2164801, IPC AC 35/74, publ. 10.04.01, including washing and sterilization of porous enterosorbent SUMS-1 modified carbon, and cooling to room temperature, the mixing of the enteric bacteria-eubiotics with nourishing and protective environments in the ratio of no greater than 1:1 and keeping the mixture for at least 1 hour under stirring at a temperature of +4°to complete immobilization of bacterial cells on the enterosorbent, washing the obtained complex preparation in isotonic saline solution followed by freezing and freeze drying.

However, used in the specified drug and the method of its production sorbents also have insufficient efficiency of sorption and desorption of bacteria-eubiotics. In addition, there is a low colonization activity included preparation of bacteria-eubiotics.

The technical result of the claimed technical solution is the creation of such a complex preparation of the probiotic with the method of its production, which would provide a higher efficiency of absorption and desorption of bacterial cells enterosorbent SMS-1 to determine its clinical efficacy, and higher colonization activity of the bacteria-eubiotics.

This technical result is achieved that, to mplexes drug-probiotic in immobilized and dried form, including the media, representing a porous enterosorbent SUMS-1 on the basis of aluminum oxide modified with carbon with a hydrophilic-hydrophobic toponimia surface, and cells of bacteria-eubiotics with nourishing and protective environments, immobilized on the specified media-enterosorbent, according to the invention the cells of bacteria-eubiotics with a nutrient medium are dried concentrate consortium of bifidobacteria or lactobacilli, or their mixture with title 108-1010CFU/g, and as modified porous carbon enterosorbent SUMS-1 it contains enterosorbent SUMS-1, optionally treated with a solution of metal salt in accordance with the method according to PP and 6 of the claims in the following ratio of components of the drug in dried form, wt.%:

The drug content of the microbial biomass is less than 0.1 wt.% has a low biotic dominated firmly bound to the sorbent cells, i.e. acts primarily as enterosorbent. The number of microbial cells introduced into the drug, does not exceed 15 wt.%, consequently all biomass of bacteria of the drug is in the protected state. This enterosorbent SUMS-1 in the product retains its basic properties, which provide the binding and excretion of intestinal toxins, products of incomplete metabolism and pathogenic microorganisms.

As a consortium of bacteria it contains strains of bacteria: Bifidobacterium bifidum 8-3, Bifidobacterium longum TWO-13 and Bifidobacterium adolescentis TH-13 in a ratio of 8:1:1.

As a consortium of lactic acid bacteria it contains strains of lactobacilli Lactobacillus acidophilus 57S, Lactobacillus plantarum P-75, Lactobacillus casei C6 in the ratio of 4:1,0:1,0.

The mixture consortia of bifidobacteria and lactobacilli has a ratio of from 1:1 to 3:1.

This technical result is also achieved by the fact that in the production method of complex preparation of probiotic in immobilized and dried form, including washing and sterilization of porous enterosorbent SUMS-1 modified carbon, and cooling to room temperature, the mixing of the enteric bacteria-eubiotics with nourishing and protective environments is in the ratio of no greater than 1:1 and keeping the mixture for at least 1 hour under stirring at a temperature not exceeding +4° With up to full immobilization of bacterial cells on the enterosorbent, washing the obtained complex preparation in isotonic saline solution followed by freezing and freeze drying, according to the invention before sterilization enterosorbent SMS-1 is treated with a metal salt solution in a volume ratio of not more than 1:1 and washed from the reagent 2-3-fold volume of distilled water, and bacteria-eubiotics use concentrate consortium of bifidobacteria or lactobacilli, or their mixture with title 108-1010CFU/g

As a solution of a metal salt use 3%solution of magnesium sulfate.

As a consortium of bifidobacteria using strains of bacteria: Bifidobacterium bifidum 8-3, Bifidobacterium longum TWO-13, Bifidobacterium adolescentis TH-13 in a ratio of 8:1:1, and as a consortium of lactobacilli used strains of lactobacilli Lactobacillus acidophilus 57S, Lactobacillus plantarum P-75 and Lactobacillus casei C6 in the ratio of 4:1,0:1,0. The mixture consortia of bifidobacteria and lactobacilli has a ratio of from 1:1 to 3:1.

The strain of bifidus bacteria: Bifidobacterium bifidum 8-3 deposited in Vniigenetika in PMBC under No 5799. Strain Bifidobacterium longum TWO-13 deposited in Vniigenetika in PMBC under No 5894. Strain Bifidobacterium adolescentis TH-13 deposited in Vniigenetika in PMBC under No 2944.

The strain of lactic acid bacteria Lactobacillus acidophilus 57S deposited in Vniigenetika in PMBC No. 583. Lactobacillus plantarum P-75 deposited in Vniigenetika in PMBC under No 3962. Lactobacillus casei C6 deposited in Vniigenetika in ACIM # 5724 In.

Introduction in the preparation of the dry microbial biomass biometra 108-1010CFU/g provides optimal biological activity. If the initial microbial biomass has a titer of less than 108CFU/g, then the product has insufficient biological activity and the introduction of the gastrointestinal tract was weak therapeutic effect. The titer of the dry microbial biomass more than 1010CFU/ml significantly increases the cost of the drug without a significant increase in treatment-and-prophylactic effect.

The basis of liquid bacterial suspension for drying are cells eubiotics in the dairy nutrient medium with a high titer of cells (109-1010CFU/ml) - liquid concentrate of bifidobacteria. Dairy nutrient medium with cells eubiotics contains increased amounts of protein (not less than 12% SOM of solids of milk). This allows you to increase the antibiotic properties of bacteria-probiotics in the product. In addition, an increased amount of protein increases antacid properties of the drug. A high concentration of cells in a liquid bacterial suspension allows them to use the drug in dry form at a concentration of 1,0-10,0% with titer of 1 8-1010CFU/ml

Media-sorbent is a mechanically durable powder particles or granules, or pellets, made of aluminum oxide, the surface of which is modified carbon upon receipt of sorbent type SMS-1 black. Sorbents have a specific surface area of up to 300 m2/g and a well-developed structure of meso - and macropores. The size of the macropores of the adsorbent commensurate with the size of microbial cells, that allows you to fill in macropores these cells. Regulation of the percentage carbon content of the sorbent and subsequent redox his treatments provides the media with the specified hydrophilic-hydrophobic and other topochemical characteristics. Sorbents type SMS-1 to a lesser extent than other similar materials, extract the vitamins and hormones from the body and do not violate peristalsis, which makes it possible to use long courses. The presence of the sorbent aluminum oxide gives its surface a certain buffer antacid properties, which contributes to a better preservation of the activity and survival of immobilized thereon microbial cells, for example, with the passage of the drug through the stomach (removes negative effect of gastric juice on microbial cells) or other unfavourable conditions (low pH).

Macro is Arista structure of the carrier-sorbent, the cells of which are filled with microbial cells, also contributes to the protection of these immobilized cells from the inactivating factors in the external environment. Moreover, microbial cells, located in the pores and cells located close to the outer surface of the carrier-sorbent, differ in their desorption abilities, and such heterogeneity properties of the cells provides a prolonged effect of the drug and promotes colonization not only the upper but also the lower intestine. Porous media, as enterosorbent, relieves the effects of local (and through him, and General toxicity, which contributes to the survival of bacteria of the preparation and of the intestinal microflora, and also reduces the load on the detoxification organs of the human or animal. In addition, when using the drug as a microbial cells consortium of bifidobacteria, or consortium of lactobacilli, or a mixture of consortia of bifido - and lactobacteria is ensured not only the displacement of the gastrointestinal tract pathogens (due to the antagonistic properties of these bacteria), but also the recovery of severely reduced intestinal microflora when dysbiosis and other diseases.

The following are examples of formulations of the complex preparation of bacteria-eubiotics in immobilized and dried form.

Example 1. Probiotic drug "Ecoflon" in the form of powder with particle size (40-100 microns) (wt.%):

cells with a nutrient medium of the consortium bifidobacteria,
or Lactobacillus-eubiotics, or their mixture
with a titer of 108-1010CFU/g1,0-10,0
protective environment0,1-10,0
enterosorbent SUMS-1, further processed
a solution of the metal saltthe rest is up to 100%.
cells with a nutrient medium of the consortium Lactobacillus
with a titer of 1010CFU/g1,0
protective environment, including:
gelatin0,085
ascorbic acid0,008
sodium chloride0,017
sucrose0,89
enterosorbent SUMS-1, further processed
3%solution of magnesium sulfatethe rest is up to 100%.

Example 2. Probiotic drug "Ecoflon" in the form of powder with particle size (40-100 microns) (wt.%):

cells with a nutrient medium of the consortium of bifidobacteria
with a titer of 109CFU/g7,0
protective environment, including:
gelatin0,85
ascorbic acid0,08
sodium chloride0,17
sucrose8,9
enterosorbent SUMS-1, additionally is processed
3%solution of magnesium sulfatethe rest is up to 100%.

Example 3. Probiotic drug "Ecoflon" in the form of granules of size (0,5-5,0 mm) (wt.%):

cells with a nutrient medium and
the titer of 108CFU/g of a mixture of consortia
bifidobacteria and lactobacilli in
the ratio of 1:110,0
protective environment, including:
gelatin0,575
ascorbic acid0,06
sodium chloride0,15
sucrose4,215
enterosorbent SUMS-1, further processed
3%solution of magnesium sulfatethe rest is up to 100%.

Example 4. Probiotic drug "Ecoflon" in the form of granules of size (0,5-5,0 mm) (wt.%):

cells with a nutrient medium and a titer of 109CFU/g
a mixture of consortia of bifidobacteria and lactobacilli in
the ratio of 3:15,0
protective environment, including the
gelatin0,85
ascorbic acid0,08
sodium chloride0,17
sucrose8,9
enterosorbent SUMS-1, further processed
3%solution of magnesium sulfatethe rest is up to 100%.

Example 5. Technology of production of ready-made forms of the drug "Ecoflon".

According to a special technology to prepare the source sorbents type SMS-1 (RF patent No. 2026733), in which the aluminum oxide modified carbon with desired porous structure, chemical nature of the surface and the size of the particles or granules. Next, measuring the number of enterosorbent SMS-1 is poured into the cylindrical container, pour distilled water in a volume ratio of 1:1 and mix thoroughly. Washed not less than 3 times with distilled water until complete disappearance of the carbon film and mist. Next enterosorbent SMS-1 is treated with a solution of metal salt, namely 3%solution of magnesium sulfate in a volume ratio of 1:1. Washed from sulfate ions 2-3-fold volume of distilled water. After processing of the enteric SUMS-1 3%-s ' solution of magnesium sulfate effectiveness of biosorption increased 1.4 times, and the effectiveness of bioresorption determining the clinical effect of the drug, equal to 1.

Sterilization SUMS-1 lead by dry heat in an oven scattering layer is not more than 1.5-2.0 cm at a temperature of 160±2)°C for 2 hours. The control of sterility SUMS-1 determined microbiologically. Control of sterilization process SMS-1 is carried out using chemical tests.

The process of immobilization SMS-1 is carried out in aseptic conditions. For immobilization take equal amounts (1:1) the number of enterosorbent SUMS-1 and culture consortia of microorganisms. Used for immobilization consortium of bacteria (with a titer of not less than 1010CFU/ml), or a consortium of lactobacilli (titer not less than 108CFU/ml), or their mixture in a volume ratio of 9:1, respectively, in accordance with examples 1-3. The biomass of bacteria are produced on a dairy nutrient medium containing high amount of protein (not less than 12% SOM of solids of milk) by standard techniques.

A mixture of enterosorbent SUMS-1 cells bacteria-eubiotics thoroughly stirred for 5 min and placed on a shaker for 1 hour at intensity of 100 oscillations per minute at a temperature of 6±2°C. the Process of immobilization may lead in a glass container on the roller installation. To determine the quality of immobilization of the original titer of bifidobacteria and lactobacilli in concentrate bifido - and lactobacteria of Wichita is t the result of biotechnologia. The process is considered complete if the solution was not more than 50% of the original quantity of bifidobacteria and lactobacilli.

Next product once washed in isotonic NaCl. A cloudy solution sterile drained, take samples and carry out the control process complete immobilization on MUK 4.2.577-96 (Methods of microbiological control of products for children, therapeutic foods and their components, p and p). The process is considered complete if the solution was not more than 50% of the original quantity of bifidobacteria and lactobacilli.

The washed product is frozen in a low temperature refrigerator type Kelvinator" at a temperature of minus (40-45)°for (24-48) hours. Cassettes with pre-frozen drug overload in the drying chamber when the temperature of the plates minus (20-30)°and a vacuum chamber. The finished product "Ecoflon" in the dry immobilized form Packed in vials and tightly stoppered. Mass fraction of moisture is not more than 3.0 wt.%.

Table 1 shows the adsorption capacity SUMS-1 after surface modification (measured by adsorption of methylene blue according FS 42-3283-96).

Table 1

Data on the adsorption capacity of the enteric SUMS-1 after treatment with metal salts
№ p/pName saltsThe adsorption capacitythe pH of the sorbent
1SUMS-1 net0,017±0.002 g8,2
2SUMS-1+AlCl3(0,36%)0,018±0.002 g6,1
3SUMS-1+AlCl3(0,75%)0,018±0.002 g5,8
4SUMS-1+AlCl3(3%)0,012±0.001 g3,26
5SUMS-1+MgSO4(1%)0,017±0.001 g6,7
6SUMS-1+MgSO4·6N2O (3%)0,024±0.002 g6,5
7SUMS-1+MgSO4·6N2About (5%)0,027±0.003 g6,3
8SUMS-1+MgCl2(1%)0,015±0.002 g6,5
9SUMS-1+MgCl2(3%)0,018±0.002 g6,6
10SUMS-1+NiCl2(3%)0,016±0.002 g5,9
11SUMS-1+ZnCl2(3%)0,019±0.001 g5,5

The treated surface SUMS-1 was evaluated in the biological tests:

1) the effectiveness of biosorption (regarding the number of adsorbed cells to the total number of bacteria in the initial suspension),

2) effektivnosti bioresorption (the ratio of the number of bacteria desorbed from the surface SUM-1, the amount of sorbed bacteria).

The effectiveness of bioresorption is an important indicator, which largely determines the clinical effectiveness of IKE. The results of the biological tests are shown in table. 2.

0,9009
Table 2

Data biological testing of enterosorbent SUMS after treatment with metal salts
Surface modificationThe efficiency. desorption LgT4/LgT3The efficiency. sorption LgT1/LgT3Source title T1 (CFU/ml)Sorption T3=(T1-T2)Desorption T4
T3T2
Immobilization of the consortium of bifidobacteria
AlCl3- 0,01%0,87141,0011·10104·1089,6·1095·108
AlCl3- 0,36%0,87811,00041·10107·107to 9.93·1096·108
AlCl3- 0,75%1,0001·10101·1089,99·1091·109
MgSO4·6N2O - 3%1,00001,0151·10103·1097·1097·109
MgSO4·6N2O - 3%0,93441,0041·10101·1099·1092·109
Immobilization mixture consortium of bifido - and lactobacteria
MgSO4·6N2O - 3%0,90681,0071·10101,6·1098,4·1091·109
Immobilization of the consortium Lactobacillus
MgSO4·6N2O - 3%0,78391,0056·1086·1075,4·1087·106
Note:

T1 (CFU/ml) - the original title of the biomass of bacteria;

T2 (CFU/ml) titer biomass of bacteria, the remainder of th is immobilization on enterosorbent;

T3=T1-T2 - titer (CFU/ml) bacterial cells immobilized on the sorbent;

T4 - titer (CFU/ml) bacterial cells after desorption, determining the clinical effect of the drug action

Thus, the results of studies of the processed surface SUMS-1 salts of metals option number 6 (table 1) processing MgSO4·6H2About (3%) is the most optimal. After processing of the enteric SUMS-1 3%-s ' solution of magnesium sulfate effectiveness of biosorption increased 1.4 times (the ratio of sorbed cells to the total number of bacteria in the initial suspension), and the effectiveness of bioresorption determining the clinical effect of the integrated product (the ratio of the number of bacteria desorbed from the surface SUM-1 to the amount adsorbed on the bacteria), equal to 1, which indicates that almost all bacteria can decorrelates in the gastrointestinal tract.

The results of preclinical studies

Studies have confirmed that the integrated product "Ecoflon" non-toxic for laboratory animals, restores normalizes intestines, eliminates conditionally pathogenic and pathogenic microflora, does not cause any adverse and unexpected Nigel the positive reactions, stimulates the process of adaptation to adverse environmental factors. Key findings:

1) morphometric characteristics of animals in the experimental groups increased in line with population standard;

2) laboratory blood tests remained within the physiological norm;

3) histological examination of the organs showed no toxic effects;

4) confirmed the absence of damaging effect on the structure of the intestinal epithelium from the drug;

5) the anatomic autopsy confirmed the organs in norm. The obtained results indicate that the integrated product "Ecoflon" successfully passed preclinical trials.

The results of clinical studies integrated product "ECOFLON"

All patients (45 people - the experimental group and 25 - man comparison group) had a chronic disease: atopic dermatitis - in 44.4% of cases; acute allergies - in 11.1% of cases; secondary chronic pyelonephritis - in 22.2% of cases, diabetes in 11.1% of cases; bronchial asthma in 11.1% of cases.

After experimental treatment complex preparation "Ecoflon" the results were as follows:

1) product "Ecoflor helps to reduce abdominal baleog the syndrome, manifestations of dyspeptic symptoms, normalization of motor function of the intestine, the regression of skin lesions;

2) the use of "Ecoflora helps to reduce subjective clinical (malaise, weakness, appetite disorder) and objective laboratory criteria (normalization of leukocyte index of intoxication and haematological toxicity, mild syndrome of cytolysis and normalization of total protein of blood serum, as well as reducing the severity of bladder syndrome), reflecting the syndrome of endogenous intoxication;

3) when use of the drug "Ecoflon" in 62.2% of the observed full recovery of the functions of digestion and absorption in the intestine, the remaining patients, a trend toward normalization of indices;

4) the drug "Ecoflon" helps to restore the intestinal microflora: Escherichia coli, bifidoflora; reduction of coccoid forms of Escherichia coli with weakly expressed enzymatic properties of lactose-negative enterobacteria;

5) if the drug is not identified side effects and complications.

Thus, the drug has successfully passed clinical trials, is harmless and highly effective means for the recovery of normalizes bowel and edema syndrome of endogenous intoxication.

1. Comprehensive probiotic drug in the immobile is organized and dried form, including the media, representing a porous enterosorbent SUMS-1 on the basis of aluminum oxide modified with carbon with a hydrophilic-hydrophobic toponimia surface, and cells of bacteria-eubiotics with nourishing and protective environments, immobilized on the specified enterosorbent, characterized in that the cells of bacteria-eubiotics with a nutrient medium are dried concentrate consortium of bifidobacteria or lactobacilli, or their mixture with title 108-1010CFU/g, and as modified porous carbon enterosorbent SUMS-1 it contains enterosorbent SUMS-1, optionally treated with a solution of metal salt in the following ratio of components of the drug in dry form, wt.%:

cells with a nutrient medium of the consortium bifidobacteria,
or Lactobacillus-eubiotics, or their mixture
with a titer of 108-1010CFU/g1,0-10,0
protective environment0,1-10,0
enterosorbent SUMS-1, further processed
a solution of the metal saltrest

2. Comprehensive probiotic drug according to claim 1, characterized in that as to what sortium bifidobacteria it contains strains of bacteria: Bifidobacterium bifidum 8-3, Bifidobacterium longum TWO-13 and Bifidobacterium adolescentis TH-13 in a ratio of 8:1:1.

3. Comprehensive probiotic drug according to claim 1, characterized in that as a consortium of lactic acid bacteria it contains strains of lactobacilli Lactobacillus acidophilus 57S, Lactobacillus plantarum P-75, Lactobacillus casei Sat in the ratio of 4:1,0:1,0.

4. Comprehensive probiotic drug according to claim 1, characterized in that the mixture of consortia of bifidobacteria and lactobacilli has a ratio of from 1:1 to 3:1.

5. The method of obtaining complex preparation of probiotic in immobilized and dried form, including washing and sterilization of porous enterosorbent SUMS-1 modified carbon, and cooling to room temperature, the mixing of the enteric bacteria-eubiotics with nourishing and protective environments in the ratio of no greater than 1:1 and keeping the mixture for at least 1 hour under stirring at a temperature not exceeding +4°to complete immobilization of bacterial cells on the enterosorbent, washing the obtained complex preparation in isotonic saline solution followed by freezing and freeze drying, characterized in that before sterilization enterosorbent SMS-1 is treated with a metal salt solution in a volume ratio of not more than 1:1 and washed from the reagent 2-3 fold volume of distilled water, and bacteria-eubiotics use concentrate konso is consortiums of bifidobacteria, or lactobacilli, or their mixture with title 108-1010CFU/g

6. The method according to claim 5, characterized in that as a solution of a metal salt use 3%solution of magnesium sulfate.

7. The method according to claim 5, characterized in that as a consortium of bifidobacteria using strains of bacteria: Bifidobacterium bifidum 8-3, Bifidobacterium longum TWO-13, Bifidobacterium adolescentis TH-13 in a ratio of 8:1:1.

8. The method according to claim 5, characterized in that as the consortium's use of lactobacilli strains of lactobacilli Lactobacillus acidophilus 57S, Lactobacillus plantarum P-75 and Lactobacillus casei Sat in the ratio of 4:1,0:1,0.

9. The method according to claim 5, characterized in that the mixture of consortia of bifidobacteria and lactobacilli has a ratio of from 1:1 to 3:1.



 

Same patents:

FIELD: medicine, surgery.

SUBSTANCE: the present innovation refers to treating osseous cavity in patients with chronic post-traumatic osteomyelitis, so, after necrsequestrectomy one should irrigate an osseous cavity with aqueous solution of sodium chloride, then it is necessary to treat the cavity with 3%-hydrogen peroxide solution with subsequent removal of foam with 0.02%-chlorohexidine bigluconate aqueous solution. Then twice during 1 min it is necessary to treat osseous cavity with 0.9%-ozonized sodium chloride solution at ozone concentration being 9000-10000 mgl/l. After that, one should introduce the mixture of anti-infectious chemotherapeutic preparations along with ultrasound cavitation for 3 min at frequency of 40 kHz followed by treating the cavity with the flow of nitrogen monoxide for 5 min at concentration being 5000 mg/cu. m. The innovation enables to increase penetrating capacity of medicinal preparations due to matching the desired mode of cavitary treatment.

EFFECT: higher efficiency of antiseptic treatment.

1 ex

FIELD: biotechnology, molecular biology.

SUBSTANCE: invention proposes a polynucleotide VEGI-192a encoding polypeptide that inhibits growth of human vascular endothelial cells. Invention describes expressing vector comprising polynucleotide and E. coli cell-host comprising vector. Invention discloses polypeptide encoded by polynucleotide and fused protein based on indicated polypeptide. Invention describes polynucleotide encoding fused protein and expressing vector based on indicated polynucleotide. Invention discloses a pharmaceutical composition used for inhibition of angiogenesis based on polypeptide-inhibitor of growth of human vascular endothelial cells and polynucleotide encoding its. Invention describes therapeutic methods for inhibition of angiogenesis and suppression of tumor growth based on this composition. Invention describes an antibody raised to polypeptide that inhibits growth of human vascular endothelial cells. Using this invention provides novel forms of inhibitor of human growth of vascular endothelial cells and can be used in medicine.

EFFECT: valuable biological and medicinal properties of inhibitor.

27 cl, 27 dwg, 13 tbl, 34 ex

FIELD: medicine.

SUBSTANCE: method involves opening pyo-inflammatory focus. Rexod is introduced in addition to basic therapy. Rexod is introduced during the operation or immediately after the operation and then every day once a day intravenously in bolus dose. The drug is introduced during 4-5 days in physiologic saline or in 5% glucose solution at a dose of 0.2 mcg/kg of patient body weight.

EFFECT: enhanced effectiveness of treatment; improved general health state within a short period; reduced oxidation stress; prevented secondary active oxygen forms production.

3 tbl

FIELD: medicine.

SUBSTANCE: disclosed is application of Ximedon (N-(β-oxyethyl)-4,6-dimethyl-1,2-dihydro-2-oxopyrimidine) for treatment and prophylaxis of various intoxication.

EFFECT: preparation of large spectrum of action and inducing effect comparable with Phenobarbital effect.

2 tbl, 13 ex

FIELD: medicine.

SUBSTANCE: method involves treating root canals with instruments and preparation containing calcium hydroxide is temporarily placed into the canals several times changing the portion every 2-5 weeks for fresh one. Before introducing fresh portion, the root canals are treated with antiseptic solution in combination with their ultrasonic treatment.

EFFECT: enhanced effectiveness of treatment; stopped inflammatory responses in periodontium tissues; prevented reinfection.

8 cl

FIELD: medicine, pulmonology.

SUBSTANCE: it is necessary to study initial values of functional reserve ability (FRA) of pulmonary-capillary circulation in %, average pressure value in pulmonary artery (AvPPA) in mm mercury column and daily variability of peak volumetric expiration rate (ΔPVRexp.) in % to calculate the following equation: D=+1.376·FRA-2.087·AvPPA-1.023·ΔPVRexp. At D value being above -25.71 one should predict instable flow of bronchial asthma. The innovation enables to carry out integral evaluation of functional state of pulmonary microcirculation, pressure in pulmonary artery and reactivity of respiratory tract.

EFFECT: higher efficiency and accuracy of prediction.

2 ex

FIELD: pharmaceutical chemistry.

SUBSTANCE: invention provides water-soluble camptotecine derivatives showing excellent therapeutical activity and retarded release suitable for cancer chemotherapy. High-molecular weight camptotecine derivative has structure wherein carboxylic acid group of block copolymer of polyethylene glycol and polymer containing carboxylic acid group in side chain and being acidic amino acid polymer is linked to phenol hydroxyl group of phenolic camptotecines through ester linkage. Also claimed is high-molecular weight camptotecine derivative of general formula (I): . Preparation method comprises combining carboxylic acid group of above-defined block copolymer with phenol hydroxyl group of phenolic camptotecines through ester linkage in presence of condensation agent. Anticancer drug includes above-defined camptotecine derivative and optionally pharmaceutically acceptable carrier.

EFFECT: augmented choice of anticancer drugs.

9 cl, 2 dwg, 2 tbl, 5 ex

FIELD: medicine, pharmacy.

SUBSTANCE: invention relates to a method for treatment of nicotine dependence in a person using a medicinal formulation with controlled release (MFCR) comprising 5,8,14-triazatetracyclo[10.3.02,11.04,9]hexadeca-2(11),3,5,7,9-pentaene or its pharmaceutically acceptable salt taken in the effective dose, and to a method for reducing adverse effect of this active component. Method involves using MFCR that comprises agents of this active component for its administration to a patient at the rate less about 6 mg/h resulting to administration of at least about 0.1 mg of compound or its salt for 24 h. The initial administration of MFCR results to plasma maximal concentration (Cmax) of active component in average from 10% to 80% of the corresponding Cmax determined for the equal dose of active component in form of bolus with immediate release, and to increasing time of value Cmax in plasma (Tmax) in initial administration in average by 50% relatively to the corresponding Tmax value determined for the equal dose of active component in form of bolus of immediate release. Also, MFCR releases active component in vireo at the rate less 6 mg/h in the dissolving testing using device USP-2 (USA Pharmacopoeia) in order to provide the dissolving time of 50 wt.-% of active component from 1 to 15 h. Indicated agents comprise a tablet with matrix, multi-particles, covered multi-particles or tablet with a cover. Invention provides carrying out the effective treatment of nicotine dependence and without symptoms of adverse effects of active component, in particle, without nausea.

EFFECT: improved method of treatment.

7 cl, 19 tbl, 13 ex

FIELD: medicine; physiotherapy.

SUBSTANCE: method can be used for doing medicinal procedures in hospitals, sanatoria and resorts. Method is based upon preparation and usage of fluid containing biological active matters made of antlers of maral. Patient's body is subject to influence of vapor at temperature of 30°-85°C. Initially water vapor is applied during 5-30 minutes. Vapor has active matters produced when vapor makes contact with herbs or herbs collection and/or aromatic oil. Then patient's body is subject to influence of vapor during 5-30 minutes, which vapor contains fluid in form of aerosol, which aerosol contains biologically active matters produced from antlers of maral. For taking procedure, vapor cabin is used, which cabin is made of two tightly joined sections. Sections are connected together along one side of joint. One section is motionless and it is connected with aroma-phyto-vapor-former by vapor duct. Cabin is provided with opening intended for patient's head. One part of opening is made in part of top horizontal surface of cabin, which part forms movable section together with part of side surface of cabin. Cabin is also provided with seat for patient. Cabin is made in form of barrel provided with vertical axis. Movable section is connected with motionless section along top side of joint and it is mounted for hoist and it keeps to vertical position by means of gas elevator. Container for biological fluid, containing biologically active matters achieved from antler of maral, is connected with device for high-dispersion spraying of fluid. Fluid communicates with cabin by means of pipeline.

EFFECT: improved efficiency of procedures due to usage of biologically active matters.

9 cl, 3 dwg

FIELD: biotechnology, immunology, biochemistry, medicine.

SUBSTANCE: invention proposes peptide concatemer inducing production of antibodies against apolipoprotein B-100 that inhibit lipase effect and inhibit binding LDL with LDL receptors. This concatemer consists of amino acid sequence of peptide repeating four times. Amino acid sequence is given in the invention description. Also, invention describes a concatemer-base vaccine used in treatment and prophylaxis of obesity and a method for preparing concatemer in E. coli cells using a vector. Invention discloses a polynucleotide encoding concatemer and expressing vector comprising the indicated polynucleotide. Using the invention provides inhibition of obesity.

EFFECT: valuable medicinal properties of concatemer and vaccine.

7 cl, 16 dwg, 1 tbl, 6 ex

FIELD: biotechnology, microbiology, medicine.

SUBSTANCE: invention relates to novel probiotic strains of lactobacilli: Lactobacillus acidophilus DSM 14869, Lactobacillus paracasei DSM 14870, Lactobacillus acidophilus DSM 15525, Lactobacillus plantarum DSM 15524, Lactobacillus sp. DSM 11526 and the strain Lactobacillus sp. DSM 15527. Strains shows similar favorable properties used separately or in combination as probiotics or in common with prebiotic as symbiotic. Also, invention relates to pharmaceutical compositions, dairy foodstuffs, functional foodstuffs, nutrient supplements and agents for personal hygiene comprising strains in separately or in combination, and to using strain for prophylaxis or treatment of vaginal infections, urogenital infections and gastrointestinal diseases.

EFFECT: valuable properties of strains.

43 cl, 12 ex

FIELD: medicine, immunology, in particular method for prophylaxis of post-vaccinal complications in infants.

SUBSTANCE: claimed method includes determination of immunoglobulin A level in infants of one or more years old, and if said level is less than 0.25 g/l vaccination is carried out and ribomunil as administered daily for 12 days.

EFFECT: decreased complication frequency in postvaccinal period.

3 ex, 1 tbl

FIELD: medicine.

SUBSTANCE: method involves treating root canals with instruments and preparation containing calcium hydroxide is temporarily placed into the canals several times changing the portion every 2-5 weeks for fresh one. Before introducing fresh portion, the root canals are treated with antiseptic solution in combination with their ultrasonic treatment.

EFFECT: enhanced effectiveness of treatment; stopped inflammatory responses in periodontium tissues; prevented reinfection.

8 cl

FIELD: medicine, dermatology.

SUBSTANCE: invention proposes a method for treatment of cutaneous wound that involves using ribomunyl aqueous solution in the dose 0.001 ± 0.0005 mg. The preparation is administrated by intradermal route in 4-5 points, not less, around cutaneous wound every day for 3-5 days. Method provides accelerating healing of cutaneous wound and formation of the more gentle connective scar. Invention can be used in treatment of cutaneous wound.

EFFECT: improved method of treatment.

2 dwg, 1 tbl

FIELD: biotechnology, microbiology.

SUBSTANCE: invention relates to a curative-prophylactic agent comprising the following components: spore biomass of microorganism-probiotics of genus Bacillus immobilized on agar-agar with a titer value 108 CFU/g, not less, condensed maize extract and microalga Spirulina platensis as a source of proteins, carbohydrates, macroelements and trace elements, and special additives in the following quantitative content of components, wt.-%: microalga Spirulina platensis, 0.5-1.5; condensed maize extract, 50-75; special components, 9.0-17.0; spore biomass of microorganism-probiotics of genus Bacillus with a titer value 108 CFU/g immobilized on agar-agar, the balance, up to 100%. Biomass of microorganism spores of genus Bacillus comprises the strain Bacillus subtilis VKPM B-7048, and/or the strain Bacillus subtilis VKPM B-7092, and/or the strain Bacillus licheniformis VKPM B-7038. Invention provides expanding assortment and increasing biological activity of curative-prophylactic probiological agent, simplifying technology for its preparing and used based on using microorganism-probiotics in spore form and without using stabilizing agents for retaining their biological activity during steps for preparing the preparation, its storage and using.

EFFECT: improved preparing method, valuable properties of agent.

8 cl, 1 tbl, 10 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes the strain Bifidobacterium bifidum 79-31 that is isolated from bowel content of health nurseling baby and this strain is multiplied actively in nutrient medium with accumulation of industrial biomass for short (12-24 h) culturing period and with the high concentration of bifidobacteria. The strain Bifidobacterium bifidum 79-31 possesses avid-forming activity, antagonistic activity with respect to pathogenic and opportunistic microorganisms and able to ferment milk. Invention provides normalization of the human body microbiocenosis being among them digestive and urogenital tracts, cutaneous and mucosa integuments and expands assortment of similar agents also.

EFFECT: improved and valuable properties of strain.

2 tbl, 5 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes the strain Bifidobacterium longum GV-15 that is isolated form the bowel content of health adult man and this strain is multiplied in different nutrient media with accumulation of industrial biomass and with the high concentration of bifidobacteria. The strain Bifidobacterium longum GV-15 possesses acid-forming activity, antagonistic activity with respect to pathogenic and opportunistic microorganisms and able to ferment milk. Invention proposes normalization of the human body microbiocenosis being among them digestive and urogenital tracts, cutaneous and mucosa integuments and expands assortment of similar agents also.

EFFECT: improved and valuable properties of strain.

2 tbl, 5 ex

FIELD: biotechnology, microbiology, food processing, medicinal and cosmetic industry.

SUBSTANCE: invention proposes the strain Bifidobacterium bifidum 79-32 that is isolated from the bowel content of health nurseling baby, and this strain is multiplied actively in nutrient media with accumulation of industrial biomass for short (12-24 h) culturing period and with the high concentration of bifidobacteria. The strain Bifidobacterium bifidum 79-32 possesses acid-forming activity, antagonistic activity with respect to pathogenic and opportunistic microorganisms and able to ferment milk. Invention provides normalization of the human body microbiocenosis being among them digestive and urogenital tracts, and expands assortment of similar agents also.

EFFECT: improved and valuable properties of strain.

2 tbl, 5 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes the strain Bifidobacterium bifidum 79-30 that is isolated from the stomach content of health nurseling baby and this strain is multiplied actively in nutrient medium with accumulation of industrial biomass for short (12-24 h) culturing period and with the high concentration of bifidobacteria. The strain Bifidobacterium bifidum 79-30 possesses acid-forming activity, antagonistic activity with respect to pathogenic and opportunistic microorganisms and able to ferment milk. Invention provides normalization of the human body microbiocenosis being among them digestive and urogenital tracts, cutaneous and mucosa integuments and expands assortment of similar agents also.

EFFECT: improved and valuable properties of strain.

2 tbl, 5 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes the strain Bifidobacterium longum ABD-6 that is isolated from the stomach content of health adult man, and this strain is multiplied in different nutrient media with accumulation of industrial biomass with the high concentration of bifidobacteria. The strain Bifidobacterium longum ABD-6 shows acid-forming activity, antagonistic activity with respect to pathogenic and opportunistic microorganisms and able to ferment milk. Invention provides normalization of the human body microbiocenosis being among them digestive and urogenital tracts, cutaneous and mucosa integuments, and expends assortment of similar agents also.

EFFECT: improved and valuable properties of strain.

2 tbl, 5 ex

FIELD: biotechnology, microbiology, medicine.

SUBSTANCE: invention relates to the strain Lactobacillus paracasei CNCM I-2116 used for diarrhea prophylaxis causing by pathogenic microorganisms. Supernatant of this strain culture elicits ability to prevent colonization of intestine with pathogenic microorganisms causing diarrhea also and this strain is designated for preparing agent used for prophylaxis and/or treatment of disorders associated with diarrhea. Agent for oral administration represents therapeutically effective dose of the strain L. paracasei CNCM I-2116 or supernatant of its culture and acceptable foodstuff. Invention provides the enhanced viability of the strain in its applying and effectiveness in prophylaxis of adhesion to intestine cells and invasion to intestine cells of pathogenic microorganisms causing diarrhea.

EFFECT: valuable medicinal properties of strain.

5 cl, 8 dwg, 10 ex

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