Method of treating radiation, chemical and/or biological damage to organism and method of obtaining globulins for treatment of radiation, chemical and/or biological damage to organism
SUBSTANCE: group of inventions relates to veterinary and medicine, in particular to obtaining and application of biopreparations for immunotherapy of ecopathologies. The group of inventions includes obtaining a protein antigen from a mixture of anatoxins from three enteropathogenic virulent strains of a pathogen of escherichiosis of calves E.coli No. 378, 379, 380 by their growing on Hottinger medium with further addition of 0.4-0.5% formalin, following thermostating for 10-12 days and cooling the mixture of anatoxins with a sterile solution of aluminium hydroxide, after that obtaining radioantigen from E.coli "PL-6" by growing cultures on a meat infusion agar with further washing away a biomass by a physiological solution and irradiation of the obtained suspension with a concentration of 1.2·1010 m.c./cm3 on a gamma-installation in a dose of 140-150 Gr with following thermostating and extraction of radiotoxin with 70% acidified with 0.05% hydrochloric acid to pH 5.5 ethanol, following evaporation of the exractant to the initial volume, after that, obtaining protein-cadmium radioantigen first by preliminary dechlorination of cadmium chloride, obtaining cadmium hydroxide, addition into the obtained 2.7% solution of the antigen of 0.77% solution of cadmium hydroxide in a ratio of 1:1 with following thermostating at a temperature of 37°C for 30 minutes, evaporation and dissolution of the residue to 12.8% concentration, further obtaining protein-cadmium radioantigen by an addition of 1.2% solution of a mixture of three anatoxins of E.coli and 12.8% solution of cadmium radioantigen in a ratio of 1:9, conjugation of components at room temperature for 8-10 hours, standardisation by a dry substance to 10% concentration and pouring into vials, and storage at a temperature of 4-6°C. The group of inventions also relates to a method of treating radiation, chemical and/or bacteriological damage to an organism by introduction of 10% solution of complex protein-cadmium radioantigen.
EFFECT: application of the group of inventions is effective in treatment of radiation, chemical and/or biological damage.
4 cl, 6 ex
The invention relates to veterinary science and medicine, in particular for the production and use of medicines for ekstraimmunnaya therapy for radiation-chemical-biological damage to the body or each of the lesions.
There is a method of treatment of radiation injuries of the body, introducing a radioprotective serum mammal subcutaneously at a dose of 100-125 mg/kg of body weight of young and 200-250 mg/kg adults within 10 days after irradiation, and a method of producing drug for the treatment of radiation injuries of the body by 2-fold the exposure of the animal donor on gamma-install, blood collection, separation of serum and subcutaneous insertion (patent RU №2169572, A61K 35/28, publ. 27.06.2001,).
The disadvantage of this invention is the limited application of the drug - protects against radiation injuries of the body.
Known associated vaccine for prophylaxis of mixed infectious diarrhea of newborn piglets, ethnological agents which are Rota-and coronaviruses and enteropathogenic strains of Escherichia coli containing antigens To 88, 99 and 987 R (patent RU №2137499, A61K 39/295, publ. 20.09.1999,).
The disadvantage of this vaccine is also limited use - only for the prevention of infectious and viral diseases of animals.
Known for the manual treatment of radiation-cadmium destruction of the body by radiation and cadmium by subcutaneous three times the introduction of radiation treatment-and-prophylactic immunoglobulin at a dose of 50 mg/kg and the introduction in the diet bentonite is based on 2% of its mass (see abstract. LR Faturahman "Combined lesions of animals Gama-radiation and cadmium and application of therapies". - Kazan, 2008. - 23 S.).
The disadvantage of this method is the absence of radiation effects - the method does not protect animals from destruction of the organism biological agents, in particular from pathogen escherichiosis.
The objective of the invention is the creation of a unified agent with a broad spectrum of therapeutic action as by radiation, chemical and/or biological damage to the body, and each lesion separately or in combination.
The problem is solved in that a method of treatment of radiation chemical and/or biological damage to the body provides for the introduction of a 10%aqueous solution of complex protein-cadmium radioantenna consisting of a mixture of anatoxina three virulent strains of E. coli No. 378, 379, 380 and protein-cadmium radioantenna, which is 4 times subcutaneously injected animals receive hyperimmune anticigarette, then separated from her globulins, which is injected once subcutaneously at a dose of 20-25 mg/kg for protein affected animals. And 4-fold subcutaneous injection of a 10%aqueous solution of complex protein-cadmium radioantenna conducted at intervals of 2 weeks - 1-I, 2-I, 3-I injection, then every 4 weeks 4-I injection at a dose of 1 mg protein-cadmium radioantenna + 500 ál complete adjuvant's adjuvant for the 1st and 2nd injection in a dose of 1 mg protein-cadmium radioantenna + 500 μl of physiological saline for 3-rd and 4-th injection, and after 7 days of taking blood samples and the resulting antisera are tested in indirect variant of ELISA for the presence of anticariogenic, anticaries and antirationalistic polyclonal antibodies, and then one of them secrete globulins by vysalivaniya ammonium sulfate.
The problem is solved and that created a method of obtaining globulin for the treatment of radiation, chemical and/or biological destruction of the body, providing first receiving protein antigen from a mixture of anatoxina of three enteropathogenic virulent strains of the pathogen escherichiosis calves E. coli No. 378, 379, 380 by growing them on the environment Hottinger followed by the addition of 0.4-0.5%formalin, further temperature control at 37°C for 10-12 days and depositing a mixture of anatoxina sterile solution of hydroxide of aluminum, then get radioantenna from E. coli "SQ-6" by growing the culture on mesopatamia agar for 48 hours, followed by washout of biomass saline solution and irradiating the resulting suspension with a concentration of 1.2-1010M.K./cm3on gamma-installation doses is 140-150 Gr with further temperature control within 3.5-4.5 h at 37°C and extraction of radiotoxins 70%acidified 0,05% hydrochloric acid to pH 5.5 ethanol, followed by evaporation of the solvent on a rotary evaporator at a temperature of 20°C up to the original volume, and then obtaining protein-cadmium radioantenna first pre-dechlorination of cadmium chloride, cadmium hydroxide, and then received a 2.7%solution of radioantenna add 0,77%solution of cadmium hydroxide in the ratio of 1:1 with further temperature control at 37°C for 30 min, evaporation and dissolving the precipitate to 12.8%concentration, further obtaining protein-cadmium radioantenna by adding 1.2 percent-aqueous solution mixture of three anatoxina E. coli and 12.8%-aqueous solution of cadmium radioantenna in the ratio of 1:9, further conjugation of the components at room temperature for 8-10 hours, standardization of dry matter up to 10%concentration and further Packed in vials and store at a temperature of 4-6°C. And dechlorination of cadmium chloride is carried out by alkaline hydrolysis using, for example, sodium hydroxide in the ratio of 1:2, respectively, is heated to a temperature of 40-45°C, the hydrolysis of lead during the night, the supernatant liquid containing dissolved Sodium chloride, decanted, and received the Zadok the cadmium hydroxide diluted to 1%concentration and is used to produce protein-cadmium radioantenna.
Obtained according to the described technology globulins provide effective treatment of injuries of the body, caused when a comprehensive radiological, chemical and/or biological effects, and their separate effects.
Technology of production of globulin for the treatment of radiation, chemical and/or biological destruction and treatment of radiation, chemical and/or biological destruction of the body consists of 4 stages.
At stage 1 prepare the protein part of the complex of antigen esherihioznae toxoid (EAT). To obtain use 3 virulent (enteropathogenic) strain of the pathogen escherichiosis calves: E. coli No. 378, 379 and 380, which are grown on the environment Hottinger for 48 h at 37°C followed by the addition of formalin (0.4-0.5 percent), the temperature control at 37°C for 10-12 days and depositing a mixture of anatoxina sterile solution of aluminium hydroxide (GOA) at the rate of 1% on the entire amount.
After settling toxoid within 2-3 days, the supernatant decanted, the precipitate + a mixture of anatoxins in the form of a gel is used as a protein carrier heptenophos part of the conjugated antigen (cadmium-plated derived radioantenna).
In the second stage tehnologicheskogo the loop gain microbial radiotoxic (RT).
As a donor antigen used vaccine strain of E. coli DPS-6, which are sown on solid nutrient medium (MPA) and grown under aerobic conditions for 48 h at 37°C, then the biomass is washed off with saline pH of 7.2, washed three times by centrifugation, centrifugal resuspended in physiological solution at a concentration of 1.2-1010M.K./cm3and use to highlight RT. For this purpose, the microbial suspension with the specified density is irradiated with gamma set to "Researcher" in a dose of 140-150 Gr with subsequent temperature control culture for 4 h at 37°C. after the specified exposure of microbial biomass precipitated by centrifugation and the precipitate is added 96% ethanol in the ratio 1:5 and extracted at room temperature for 2 hours and Then the extract was centrifuged at 8000 rpm for 30 min, the supernatant is poured.
The obtained extract was evaporated on a vacuum rotary evaporator to original volume and neutralized with 0.1 n KOH to a pH of 7.4, diluted with saline to 2.7%concentration (27 mg/ml) and used in future work as one of the components (radioantenna) politicheskogo complex.
At the third stage receives cadmium derived radioantenna - quinoid radiotoxins. At the same time to reduce the toxicity of the complex and is Tigana previously conducted dechlorination of cadmium chloride (CdCl 2) by alkaline hydrolysis using NaCl in the reaction:
To do this, prepare aqueous solutions of 1-molar solution CdCl2(182 g/mol) and 2 molar NaOH solution (149,12 g/mol), which are in the ratio 1:2 (for example, 100 ml CaCl2and 200 ml of NaOH) is drained, bring the pH to 7.7 to 8.0, heated to 35-40°C and hydrolysis are during the night, and then the supernatant liquid containing dissolved Sodium chloride, decanted and the precipitate - cadmium hydroxide (Cd(OH)2) diluted to a concentration of 1% solution (10 mg/ml) and used to produce derived radioantenna.
Radioaktiven representing the chemical composition of the oxidized quinone (o-quinone), which is quite highly reactive active radical free (broken) double bond in the oxidation process that can easily attach ions organic (protein, amino acids, enzymes and inorganic (metals and other chemical agents) substances. In turn, the cadmium hydroxide is a weak electrolyte in aqueous solutions it is hydrolyzed ions (Cd+, OH) and active Cd+easily attaches to the oxidized o-quinone at the place of rupture of the double bond, forming a cadmium derived quinone C6H10OCd.
To obtain cadmium derived radioantenna (CPRA) to a specific volume (e.g., 5 ml) of 2.7%solution of radioantenna add equal volume of 0.77%-aqueous solution of cadmium hydroxide (Cd(OH) 2) and the mixture thermostatic when shuttlebay for 30 min at 37°C. Then the solvent is evaporated in a vacuum centrifuge. The residue was dissolved in minimum amount of water and added to the protein solution (toxoid, E. coli)used as a carrier heptenophos part of polyanthea - cadmium radioantenna. The amount of cadmium radiotoxins is 10-fold molar excess relative to the amount of protein: to 50 ml of a 1.2%solution toxoid E. coli add 50 ml of 12.8%-aqueous solution of cadmium radioantenna.
The conjugation reaction is carried out in over night at room temperature.
To confirm the stability of the conjugated antigen in solution, measuring the molar ratio of cadmium, radioantenna and protein in the conjugate conduct spectral analysis by measuring optical densities of native components antigens (CdCl2o-quinone and toxoid) and the conjugated antigen on the spectrophotometer.
To measure the molar ratio of cadmium derived o-quinone (radioantenna) and protein using the method of Bradford (Bradford reagent). For this kumassi brilliant blue G-250 (100 mg) is dissolved in 50 ml of 95% ethanol. Then add 100 ml 85% (mol. about.) phosphoric acid, dilute with water to 1 L. Use the sample containing 5 μg of protein in 1 ml. In the cell we use the t 0.5 ml of reagent dye (Bradford reagent), then make 1 μl of sample, mix well. After 10 minutes measure the optical density at 595 nm.
The protein concentration is determined according to a calibration curve constructed using endotoxin of E. coli. Then the spectrum of the conjugate to determine the molecular concentration CdCl2, o-quinone, and then determine the molecular ratio CdCl2, o-quinone and protein in the conjugate. The lack of trianthema complex absorption spectra characteristic of the individual components constituting a single trencheny complex, indicates the stability of the conjugate.
Obtained with the aforementioned method trencheny complex - protein-cadmium radioaktiven (BKRA), representing a 10%solution of antigens containing 1 ml 100 mg antigenic substances, of which the share of toxoid have 10 mg, the share of radioantenna - 78 mg and at a fraction of the cadmium - 12 mg, the following 4-th stage is used as an immunizing agent to obtain antisera. To exclude toxic effects BKRA on the body of the source (10%) solution of antigen diluted in 50 times, thus obtaining 0,2%working solution, 1 ml of which contains 2 mg of antigenic substances, of which, according to the above mixing ratio, the share of ecotoxins protein is necessary to 0.2, mg, radioantenna - 1.56 mg is cadmium - 0,24 mg
To determine the antigenic (antitelomerase) activity of conjugated antigen (BKRA) use of white mice, which are subjected to immunization by 4-fold scheme. The first immunization is carried out by subcutaneous injection of 1 mg of conjugate in 500 ál of surfactants, 2nd 2 weeks 1 mg conjugate in 500 μl of surfactant, 3rd 2 weeks 1 mg conjugate in 500 μl saline, 4th after 4 weeks of 1 mg conjugate in 500 μl of saline.
To test antisera week after the 4th immunization, the animals are selected based on 50 μl of blood from the jugular vein.
Given that the optimal method of determining the quantity of antibody is enzyme-linked immunosorbent assay (ELISA), and more affordable and easy option - indirect competitive method, when testing antisyware use the latest version of the test system.
For carrying out the indirect competitive ELISA analysis prepared immunological tablets. To do this, prepare working conjugate solution BKRA with a concentration of 10 μg/ml In each well of the tablet make 50 μl of the obtained solution and incubated in a shaker-incubator for 1 h at 37°C, 500 rpm Washed tablet 3 times with phosphate-saline buffer solution (FSB) pH to 7.2 with 0.05% of Tween-20. Then, to prevent nonspecific adsorption of reagents, contribute to each well of the tablet 100 μl of a 2%aqueous solution ekzotik the ina (toxoid) E. coli in the FSB and incubated for 30 min at 37°C, 500 rpm After that, the tablet is washed 5 times with a solution of the FSB (pH of 7.2) with 0.05% of Tween-20.
In the finished tablets make 50 μl of serial two-fold dilutions received hyperimmune antisera (from 1:2 to 1:1024). Each dilution bring in 2 replications (i.e., 2 wells for each dilution).
Incubated tablet 1 h at 37°C, 500 rpm in a shaker-incubator. Then washed tablet 3 times with a solution of the FSB (pH of 7.2) with 0.05% of Tween-20 and make the solution artemisinin antibodies conjugated with horseradish peroxidase, the FSB with a 1% exotoxin E. coli. Incubated 1 h at 37°C, 500 rpm in a shaker-incubator and washed tablet 5 times with a solution of the FSB (pH of 7.2) with 0.05% of Tween-20. Make a substrate for horseradish peroxidase (3,3', 5,5' -tetramethylbenzidine). Incubate 10 min at room temperature, then stop the color reaction by adding an equal volume (50 ál) of 1M sulphuric acid (H2SO4). In tablet spectrophotometer record the absorbance at 450 nm, the titers obtained sera depending on the time of immunization is 1:64-1:500.
To confirm the specificity of the obtained antisera against the free toxoid, E. coli, radiotoxic and Ecotoxicity - cadmium chloride they are examined by the method of competitive ELISA, which examine the interaction of serum-free anti-Christ. s, components of the conjugated antigen (BKRA). This serum at selected breeding contribute to the wells of the prepared microplate with different dilutions of free exotoxin (toxoid) E. coli, radiotoxins (radioantenna) and cadmium chloride. They inhibit the interaction of sera with conjugate BKRA, sorbed on the solid phase (i.e. antibodies interact with free exotoxin E. coli, radiotoxins and cadmium chloride), as judged by the change in optical density at 492 nm. The decrease in optical density from 0.7 to 0.3 (adding toxoid, E. coli) from 0.5 to 0.05 (when adding radioantenna) and from 0.45 to 0.12 (adding CdCl2) indicates the presence in the tested sera specific anticariogenic, antirationalistic and anticaries antibodies, which are able to interact with free immunotoxic agents - exotoxin E. coli, radiotoxins and ecotoxicants - CdCl2in the body, to neutralize them, providing, thus, antitoxic effect as a stand-alone and combined entering them in the body.
Thus, the antisera obtained by immunization of animals with complex polyanthemos (BKRA)are polyclonal antibodies with different immunological competence with the ability to join the chemical bond with ecotoxicants (radiotaxi, exotoxins, heavy metals), which is the basis for testing them as a means ekstraimmunnaya therapy, with the defeat of the organism by physical agents (radiation), chemical (ecotoxicants - CdCl2) and biological (E. coli) nature.
For standardization immunochemical structure of antibodies and increase their therapeutic activity of antisera isolated globulin in accordance with generally accepted immunochemistry methods (see kN. "Experimental immunochemistry". Ed. by N.V. Kolcheva. - M.: Medicine. -1968. - 665), globulin precipitate was diluted to 10%concentration (100 mg/ml), Packed in sterile glass bottles of 500 cm3, sterilized and stored in a refrigerator at 4-6°C.
Received 10%solution of globulins administered irradiated suffering from colibacillosis and affected cadmium chloride animal once subcutaneously in doses of 20-25 mg/kg) protein (0.4 ml/kg).
A method of obtaining a conjugated antigen and multifunctional therapeutic globulin is illustrated by the following examples.
Example 1. To obtain esherihioznae toxoid virulent strains of enterotoxigenic E. coli No. 378, 379, 380 were seeded into a two-liter flask with the environment of Hottinger (pH 7.8), were grown for 2, 3, 4, 5, 6, 7, 8, 9 days at a temperature of 35, 36, 37, 38, 39°C. Then the culture was added formalin to a concentration of 0,1, 0,2, 0,3, 0,4, 0,5, 0,6 and 0.7%, withstood thermostate at 35, 36, 37, 38°C for 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 days. The resulting toxoid besieged 5, 7, 9, 10, 12, 14%-tion sterile solution of aluminium hydroxide (GOA) based 0,5, 0,7, 0,9, 1,0, 1,1, 1,2% on the whole volume. The results of the experiments showed that the most high biological activity has a mixture of anatoxina obtained from three strains of E. coli No. 378, 379 and 380, grown on the medium Hottinger for 24 h followed by the addition of formalin rate of 0.4-0.5% and a temperature of the cultures at 37°C for 10-12 days, and deposition of a mixture of aluminum at the rate of 1 vol.%. Any changes incubation of cultures (temperature, exposure), the concentration of formalin and the number of strains-producers resulted in reduced biological activity of anatoxina.
Example 2. To obtain the second component of radioantenna (oxidized o-quinone), a strain of E. coli "SQ-6" were sown on solid nutrient medium (MPA) and were grown at 37°C for 24, 48, 72 hours and Then the biomass was washed with saline, centrifuged at 5000, 6000, 7000, 8000, 9000, 10000 rpm, repeating the procedure 1, 2, 3, 4, 5 times. Sediment resuspendable in physiological solution at a ratio of 1.0·1010, 1,1·1010, 1,2·1010, 1,3·1010, 1,4·1010M.K./cm3and were irradiated with gamma set to "Researcher" at doses of 50, 100, 200, 250, 300 Gy at the dose rate of 2,064 CL. After irradiation, the culture in which was aktivirovalo at 35, 36, 37, 38, 39°C for 1, 2, 3, 4, 5, 6 h and perform the extraction of radiotoxins using 50, 60, 70, 80, 90%-aqueous acidified(0,01, 0,02, 0,03, 0,04, 0,05 0,06, 0,07% HCl) to pH 5,1, 5,2, 5,3, 5,4, 5,5, 5,6, 5,7 ethanol, repeating this operation 2, 3, 4, 5 times. The obtained extracts were evaporated on a rotary evaporator at 10, 15, 20, 25°C up to the original volume and neutralized 0,05, 0,07, 0,1, 0,2, 0,3 N. KOH to pH 6,4, 6,5, 6,7, 6,8, 7,0, 7,1 and 7.2. The results of the experiments showed that the most active drugs radiotoxins (radioantenna) obtained when E. coli is grown "SQ-6" at 37°C for 24 h, the irradiation of the microbe suspension with a concentration of 1.2-1010M.K./cm3at a dose of 140-150 Gr, temperature control within 3.5 to 4.5 at 37°C, the ethanol extraction of the acidified culture (0,05 HCl) to pH 5.5 and 5.6, the evaporation of the extractant at 37°C. Any changes in these parameters in the direction of decreasing or increasing the lead to reduced biological activity of radiotoxins.
Example 3. To obtain cadmium radioantenna (quinoid RT) to reduce the toxicity of cadmium chloride and enhance its reactivity initially conducted its dechlorination by hydrolysis CdCl2a concentrated solution of alkali (NaOH). This was prepared saturated (40, 50, 60%) solutions of alkali and CdCl2and mixed them in a molar ratio (g/mol) CdCl2+NaOH 0,5:0,5, 1:1, 1:1,5, 1:2, 1:2,5, 1:3. The hydrolysis was conducted at 20, 25, 30, 35, 4°C for 10, 15, 20, 25, 30 hours of exposure. After these exposures adosados removed. It is established that the maximum yield of cadmium hydroxide which was achieved when the molar ratio of components (CdCl2+NaOH) 1:2 with 10-12 hours of exposure and temperature of 35°C.
Example 4. To obtain cadmium radioantenna to 10, 20, 30, 40, 50 ml 1,0, 1,5, 2,0, 2,5, 2,7, 2,9, 3,0%-aqueous solution of radioantenna added 10, 20, 30, 40, 50 ml 0,50, 0,60, 0,65, 0,70, 0,73, 0,77, 0,80, 0,85%-nye solutions of cadmium hydroxide (Cd(OH)2) and the mixture is incubated in a thermostat at 25, 30, 35, 37, 39, 40°C for 10, 15, 20, 25, 30, 35 minutes
The results of the spectral analysis showed that the maximum yield of the final product - cadmium radioantenna is achieved when the mixing ratio 3:1, i.e. when added to 50 ml of a 2.7%solution of 0.77%-aqueous solution of cadmium hydroxide, which was the ratio of 3:1. Any changes to the specified ratio led to a decrease in the yield of the final product.
Example 5. To obtain a conjugated protein-cadmium radioantenna (BKRA) was prepared 0,03-, 0,06-, 0,07-, 0,08-, 0,09-, 0,1-, 0,2-, 0,3-molar solutions altobella (toxoid, E. coli) and 0,5-, 0,6-, 0,7-, 0,8-, 0,9-, 1,0-, 2,0-, 3,0- molar solutions of cadmium radioantenna and 50 ml solutions of each dilution altobella were added to 50 ml of solutions of cadmium radioantenna, the conjugation reaction was carried out for 18, 19, 20, 21, 22 23, 24, 25 hours at 20, 22, 24, 26, 28, 30, 32, 34, 36°C.
After these exposures were determined by the stability of the conjugated antigen and the optimum molar ratio of components in polietilene method of spectral analysis by measuring optical densities of native components antigens (CdCl2, o-quinone and toxoid) and the conjugated antigen on a spectrophotometer (for example, SF-46). As an indicator of the stability of the obtained antigen complex was the lack of trianthema complex absorption spectra characteristic of the individual components of polyanthea, and the molar ratio CdCl2, o-quinone and altobella E. coli in polietilene was determined by measuring the optical density of solutions with different molar ratios of the individual components of the conjugate. It was established that the values of optical densities of polyanthea and individual components (Akshabulak-toxoid E. coli: radioaktiven: cadmium) coincided with molar ratios of 1:7,8:1,2, respectively.
Example 6. To determine the antigenicity (antitelomerase) activity obtained protein-cadmium radioantenna (BKRA) conducted immunization of mice. Used 2 groups of experimental animals, each of which consisted of 5 animals. The first group received polianthes NR is drybrushing, the second subcutaneously. Used 4-fold scheme of immunization with varying doses (0.25, and 0.50, and 1.0, 1.5 mg antigen) and interval (2, 4, 6 weeks) between the introduction of the conjugate.
The results of the testing of antisera live version of ELISA showed that the highest antibody titer was detected in the animals subjected to 2 scheme immunization (2nd group), suggesting a 4-fold subcutaneous administration of antigen with an interval of 2 (1-I, 2-I, 3-I injection) and 4 weeks (4th injection) in doses of 1 mg BKRA+500 ál complete adjuvant's adjuvant (PAF) (1-I injection, 1 mg BKRA+500 μl incomplete adjuvant's adjuvant (NAF) (2-I injection, 1 mg BKRA+500 μl of physiological solution (3-I and 4-I injection, respectively).
Example 7. For the evaluation of radioprotective, antisilicone and anticaries activity polyglobulia experiments put on irradiated at a dose of 7.7 G, inoculated cadmium chloride at a dose 50,8 mg/kg and infected with a pathogen escherichiosis 1·109M.K. white mice over 24 h before and 24 h after irradiation, a dose of CdCl2and infection with a pathogen escherichiosis was administered subcutaneously once globulins in doses of 0.12 (6 mg/kg)0,25 (12 mg/kg)0,5 (25 mg/kg), 1.0 mg (50 mg/kg). Before applying the original drug (10%solution globulin) were diluted in 2, 4 and 8 times, and thus, 5, 2.5 and 1.25 percent by concentration of the drug.
It is established that the maximum radioprotective (vigeveno the ü 66.6% of irradiated animals), antisilicone (survival 59,1% is infected with the causative agent of escherichiosis animals) and antidote (survival 61.9% of the inoculated CdCl2animals) occurred at a single subcutaneous injection globulin at a dose of 25 mg/kg Change in dose reduction resulted in reduced therapeutic effect, and the increase is to increase the consumption of the drug. Therefore, the optimal therapeutic dose is 25 mg/kg for protein.
Thus, conjugated designed polyantigenic the drug produces polyclonal antibodies - globulins with polyfunctional curative properties when ecopathology caused by agents of radiation, chemical and/or biological nature.
1. The method of obtaining globulin for the treatment of radiation, chemical and/or biological destruction of the body, providing first receiving protein antigen from a mixture of anatoxina of three enteropathogenic virulent strains of the pathogen escherichiosis calves E. coli No. 378, 379, 380 by growing them on the environment Hottinger followed by the addition of 0.4-0.5%formalin, further temperature control at 37°C for 10-12 days and cooled mixture of anatoxina sterile solution of hydroxide of aluminum, then get radioantenna from E. coli "SQ-6" by growing crops on meats is peptone agar for 48 hours subsequent washout of biomass saline solution and irradiating the resulting suspension with a concentration of 1.2·10 10M.K./cm3gamma-installing dose of 140-150 Gr with further temperature for 3.5-4.5 hours at 37°C and extraction of radiotoxins 70%acidified 0,05%hydrochloric acid to pH 5.5 ethanol, followed by evaporation of the solvent on a rotary evaporator at a temperature of 20°C up to the original volume, and then obtaining protein-cadmium radioantenna first pre-dechlorination of cadmium chloride, cadmium hydroxide, and then received a 2.7%solution of radioantenna add 0,77%solution of cadmium hydroxide in the ratio of 1:1 with further temperature control at 37°C for 30 min, evaporation and dissolution of the precipitate to 12.8%concentration, further obtaining protein-cadmium radioantenna by adding 1.2 percent-aqueous solution mixture of three anatoxina E. coli and 12.8%-aqueous solution of cadmium radioantenna in the ratio of 1:9, conjugation of the components at room temperature for 8-10 hours, standardization of dry matter up to 10%concentration and filling vials, storage at a temperature of 4-6°C.
2. The method according to claim 1, characterized in that the dechlorination of cadmium chloride is carried out by alkaline hydrolysis using, for example, sodium hydroxide in the ratio of 1:2, respectively, is heated to a temperature of 40-45°C, hydrolysis lead to mortgage the e night, the supernatant liquid containing dissolved Sodium chloride, decanted, and the resulting precipitate the cadmium hydroxide diluted to 1%concentration and is used to produce protein-cadmium radioantenna.
3. The treatment for radiation, chemical and/or biological destruction of the body, introducing a 10%solution of complex protein-cadmium radioantenna obtained by the method according to claims 1, 2, which is four times subcutaneously injected animals receive hyperimmune anticigarette, then separated from her globulins, which is injected once subcutaneously at a dose of 20-25 mg/kg for protein affected animals.
4. The method according to claim 3, characterized in that the four subcutaneous injections of 10%aqueous solution of complex protein-cadmium radioantenna conducted at intervals of 2 weeks (1st, 2nd, 3rd injection), then with an interval of 4 weeks - 4 injection, at a dose of 1 mg protein-cadmium radioantenna + 500 ál complete adjuvant's adjuvant for the 1st and 2nd injection in a dose of 1 mg protein-cadmium radioantenna + 500 μl of physiological saline for 3-rd and 4-th injection, and after 7 days after the last injection of antigen take blood samples and the resulting antisera are tested in indirect variant of ELISA for the presence of anticariogenic, anticaries and antirationalistic polyclonal antibodies, and then one of them secrete globulins way in which slyvania ammonium sulfate.
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to medicine and particularly to preparations for a therapy of oncological diseases, treatment of allergy, prevention and health improvement. The invention represents a method for preparing fraction 4 (ADP-f4) and fraction (ADP-f5) adaptogenic Dorogov's preparations, characterised by the fact that a primary product is a fraction 2 antiseptic Dorogov's stimulator (ADP-f2) to be thermally treated.
EFFECT: implementing the given invention provides the more effective prevention and eliciting anti-stress reactions for the purpose of improving the functional state of organism and increasing a resistance in an activation therapy.
SUBSTANCE: invention refers to medicine and veterinary science and aims at increasing the radioresistance in mammals. In the period of 12 hours to 20-30 minutes before a radiation exposure and immediately thereafter, 0.11% oil preparation of chlorophyll is introduced into the mammals. Linseed oil is used as an oil base. The preparation is introduced in a dose of 11-12 mg/kg intramuscularly.
EFFECT: method enables increasing the radioresistance and survival rate in the mammals.
1 tbl, 1 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: composition for treating oxidative stress comprises ball-shaped lipoic acid or one of salts thereof, and at least one lipophilic medium. The lipoic acid balls represent particles consisting of an inert core (a nucleus) coated with lipoic acid which is coated with a first layer of an isolating polymer, and with a second polymer layer resistant (stable) at gastric pH. What is also described is a preparation for treating oxidative stress with an unified dose containing the above composition. The preparation is presented in the form of a soft gelatin capsule.
EFFECT: compositions according to the invention are stable in the lipophilic medium.
22 cl, 15 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions refers to medicine, particularly toxicology and radiology, to drug preparations based on antioxidant proteins and methods of using them. The pharmaceutical composition for treating toxic conditions wherein the therapeutic effect is ensured by the action of antioxidant, antimicrobial, antitoxic human lacroferrin protein on the human body contains non-replicating nanoparticles of human adenovirus serotype 5 genome with inserted human lactoferrin expressing human lactoferrin in the therapeutically effective amount in the body, and an expression buffer with the particle content not less than 2.33×1011 of physical particles per ml of the expressing buffer. The method of therapy involves administering the composition in the therapeutically effective dose of 7×1011 of physical particles to 7×1013 of physical particles per ml of the expressing buffer per an individual; the composition is administered intravenously.
EFFECT: invention provides the stable therapeutic effect after the single administration of the composition.
17 cl, 14 ex, 4 dwg
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to medicine, and concerns a pharmaceutical composition for the therapy of acute toxic conditions. The composition for the therapy of acute toxic conditions contains a protein - human lactoferrin, and additionally contains non-replicating nanoparticles with an inserted gene of human lactoferrin, and an expressing buffer in the following proportions per a dose: human lactoferrin 50 to 100 mg; non-replicating nanoparticles - 7×1011 of physical particles; the expressing buffer - the rest, ml. Human lactoferrin is either donor breast milk lactoferrin, or any human lactoferrin.
EFFECT: invention provides fast onset and prolonged antitoxic action.
4 cl, 8 ex, 2 dwg
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to medicine and pharmacy and can be applied in production and application of solutions for intravenous introduction in treatment of conditions, associated with endogenous intoxication. Disintoxication solution contains components in the following ratio (wt %): sodium hypochlorite 0.04-0.08, sodium chloride 0.50-1.00, aminoethane sulfonic acid 0.02-1.20.
EFFECT: novel infusion solution is claimed.
4 ex, 1 tbl
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to pharmaceutical industry, namely an agent possessing anti-inflammatory action. The agent possessing anti-inflammatory action contains as active ingredient in the form of a complex prepared of a coelomic fluid of sea urchins containing peptides and amino acids at a certain ratio of the ingredients.
EFFECT: agent possesses a manifested anti-inflammatory effect.
2 cl, 2 tbl, 10 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions refers to a method for preparing a carbon sorbent with the antibacterial properties, and to the carbon sorbent with the antibacterial properties prepared by this method. The declared method involves impregnation of the carbon hemosorbent granules in an initiator solution in N-vinylpyrrolidone at pH 7.0-7.5 and a residual pressure of 15-20 mm Hg. The hemosorbent : initiator solution in N-vinylpyrrolidone ratio is 1:1.4-2.0. Then the temperature is raised to 65-75°C, kept at that temperature for 0.5-8 hours in an inert medium and washed in water from the residual monomer at room temperature.
EFFECT: carbon sorbent with the antibacterial properties prepared by the specified method represents the round granules, contains polyvinylpyrrolidone in an amount of 4,5-5,5% and is characterised by a specific surface adsorption of less than 50 m2/g and total pore volume of less than 0,30 cm3/g.
2 cl, 2 tbl, 1 dwg, 7 ex
SUBSTANCE: invention refers to medicine and aims at treating necrotic ulcers of oral mucosa in patients with multiple myeloma. Luminal-dependent chemiluminescence of oral fluid is conducted to determine fluorescence light sum and maximum flash. If the light sum value is within the range of 1.02 to 6.6 standard units, and the maximum flash value is 0.7 to 2.5 standard units, the toothpaste Lacalut fluor is used 2 times a day and solcoseryl dental adhesive paste 4 times a day after meals and before bedtime to relieve symptoms completely. The light sum value 8.3 to 13.5 standard units and the maximum flash value 3.4 to 4.2 standard units, the toothpaste Colgate total propolis is used 2 times a day and the film preparation Oblecol three times a day to relieve symptoms completely.
EFFECT: use of the invention reduces a length of epithelisation of necrotic ulcers of oral mucosa.
FIELD: medicine, pharmaceutics.
SUBSTANCE: what is presented is using water-soluble hybrid macromolecular compounds O-(3-(3,5-di-tert-butyl-4-hydroxyphenyl)propionyl-(1→6)-α-D-glucan and polyethylene glycol bis-3-(3,5-di-tert-butyl-4-hydroxyphenyl)-propionate as erythroprotective agents for phenylhydrazine poisoning. It is shown that the course intravenous introduction of the compounds has manifested erythroprotective action.
EFFECT: invention extends the range of agents applicable in integrated therapy of phenylhydrazine poisoning.
1 tbl, 3 ex
SUBSTANCE: cosmetological set comprises 25-50 biodegradable meso-sutures based on polydioxanone, attached at the sharp ends of the guide-needles of high strength steel with size of 25G-31G, and 0.75-2.5 ml of the preparation on the base of the hydrolyzate of placenta, which is a Laennec and/or Curacen, and also a method of lifting the soft tissues using the above set.
EFFECT: effective skin rejuvenation and removal of soft tissue defects with aesthetic long-lasting effect in patients of different age groups.
4 cl, 3 ex
SUBSTANCE: invention enables creating complex formulations of functional sports supplements of single foods with the specified concentrations of vitamins and mineral substances for sportsmen of various sports.
EFFECT: recovered physiological body saturation with vitamins and mineral substances on the basis of an algorithm for determining sportsmen's body saturation with these nutrients.
SUBSTANCE: compounds have the general formula: X1-X2-X3-Pro-X4-Pro-X5, where X1 is selected from H, Ac-, Palm-; X2 is selected from Trp, Tyr; X3 is selected from Trp, Tyr; X4 is selected from Lys, Orn, Dbu, Dpr, Arg; X5 is selected from-OH, -NH2, -OCH3, -OC2H5, -NH-C6H5. This compound can be represented in the form of retroisomer.
EFFECT: invention enables to block selectively the muscle nicotinic acetylcholine receptor with high efficiency.
12 cl, 7 dwg, 1 tbl, 5 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to the field of pharmaceutics. An extract of Fraxinus excelsior seeds, capable of activating PPAR-alpha, which contains nuzhenide GI3, oleoside methyl ester, excelside B, GI5, salidroside, in effective quantities. An application of the extract of Fraxinus excelsior seeds to obtain a medication for treating a state, in which the PPAR-alpha activation is useful, is described. Also described is a method of treating a subject with the state, in which the PPAR-alpha activation is useful. A method of obtaining the extract of Fraxinus excelsior seeds is disclosed.
EFFECT: application of the claimed extract makes it possible to treat states, in which the PPAR-alpha activation is useful in an efficient way.
14 cl, 11 dwg, 2 tbl, 13 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions relates to a composition for teeth cleaning and a method of its obtaining. The claimed composition for teeth cleaning contains in one phase an orally acceptable carrier; a source of fluorine ions; a source of bivalent tin ions; a source of zinc ions and, at least, one polyphosphate salt, selected from a group, which consists of inorganic polyphosphates, which have 3 or fewer atoms of phosphorus; and the general content of water in the composition for teeth cleaning constitutes less than approximately 10% of the composition mass. The method of obtaining the claimed composition includes mixing the source of bivalent tin ions with a water buffer system, adapted for chelation of bivalent tin ions in a premix, formed in this way; and combining the premix with the active components and the orally acceptable carrier.
EFFECT: combination of sources of bivalent tin, fluorine, zinc and the upper mentioned polyphosphate with a low content of water makes it possible to obtain the composition for teeth cleaning in the form of a one-phase system, which provides a possibility of an effective delivery of unstable in water active ingredients, which begin a reaction with each other in one phase.
28 cl, 2 dwg, 3 tbl, 5 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to the chemical-pharmaceutical and cosmetic industry and represents an antiperspirant composition, which includes a) an active component of an antiperspirant; b) vegetable oil in quantity 12-20 wt %, c) a primary gel-former, which includes C16-C18 saturated fatty acid in quantity 15-21 wt %, d) a secondary gel-former, selected from hydrogenated soybean oil, partially hydrogenated soybean oil, hydrocarbon of formula CnH2n+2, where n is equal to 20-100, and hydrocarbon by at least on 90% is linear, hydrogenated castor oil (castor wax) and fatty alcohol, where the composition represents a solid stick or a half-solid product with an index of peeling, constituting less than approximately 10%.
EFFECT: elaboration of the antiperspirant composition.
18 cl, 5 tbl, 1 ex
SUBSTANCE: invention is anti-cellulite liposomal agent comprising enclosed in liposomes of lecithin water-soluble extracts of southern sumac (Buxus sinica) and anise (Pimpinella anisum), enclosed in the inner phase of liposomes, and oil-soluble extract of pink pepper (Schinus terebinthifolius) enclosed in an outer casing of liposomes.
EFFECT: natural and induced lipolysis and inhibition of lipogenesis process after meal.
4 cl, 3 ex, 1 tbl, 2 dwg
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to the pharmaceutical industry, namely to a pharmaceutical composition for treatment of a diabetic ulcer. Application of the pharmaceutical composition in manufacturing a pharmaceutical preparation in the area ofan extremity or on a body surface, or in manufacturing a dressing material for treatment of the diabetic ulcer, where the pharmaceutical composition consists of an edible bee wax, prepared on the basis of the sesame oil extract of rhizome of Scutellaria - Huangqin (Huang Qin), rhizome of Coptis - Huanglian (Huang Lian), bark of Phellodendron- Huangbai (Huang Bai), earthworm and poppy capsule, taken in a specified ratio. The dressing material for treatment of the diabetic ulcer in the area of the extremity or on the body surface, where the said dressing material contains the pharmaceutical composition, consisting of: edible bee wax, prepared on the basis of sesame oil extract of rhizome of Scutellaria - Huangqin (Huang Qin), rhizome of Coptis - Huanglian (Huang Lian), bark of Phellodendron- Huangbai (Huang Bai), earthworm and poppy capsule, taken in a specified ratio. The first aid kit for treatment of the diabetic ulcer in the area of the extremity or on the body surface.
EFFECT: composition possesses an expressed healing activity in treatment of the diabetic ulcers.
10 cl, 20 dwg, 1 tbl, 13 ex
SUBSTANCE: present invention relates to a comb-shaped copolymer comprising: (A) one or more repeating units obtained from olefinically unsaturated cationic or cation-active comonomers; and (B) one or more repeating units of formula where Y is a moiety which forms part of the copolymer backbone and is obtained from a monomer selected from at least one of the following monomers: olefinically unsaturated cationic or cation-active comonomers, acrylamide monomers, one or more olefinically unsaturated hydrophilic monomers, one or more olefinically unsaturated monomers; Z is a moiety capable of forming an associate with another moiety Z or other moieties in the preparation in which the copolymer will be used, and is a hydrophobic moiety selected from alkyl, aryl, aralkyl, fluoroalkyl groups having 8-50 carbon atoms, an organosilicon group having 35-25 linked SiO moieties, and silane; and b is a bond or a moiety, linking the moiety Z with a moiety Y, and represents covalent bonds formed by at least one ester, carbonyl, amide, amine oxide, hydrocarbon, amino, ether, polyoxyalkylene groups, or a bond resulting from ionic salt bonds. The invention also describes a personal hygiene product and a composition for the personal hygiene product containing said comb-shaped copolymer.
EFFECT: obtaining a comb-shaped cationic copolymer, which provides a balance of the required properties when used in personal hygiene products and other cosmetic preparations in terms of the sensory perception thereof and the degree of deposition or retention of active ingredients.
25 cl, 13 tbl, 73 ex
SUBSTANCE: invention is a method of production of microemulsion cleaning agent of epicutaneous application, comprising mixing the nonionic surfactants with the oil components to homogeneous state, and adding water and polyols, characterised in that 40-60 wt % water is preliminary mixed with polyols, it is added into the mixture of nonionic surfactants with oil components, stirred to homogeneous state, and the remaining quantity of water is added, at that the process is carried out under vigorous stirring and at room temperature.
EFFECT: obtaining cleanser in the form of stable microemulsion that has the advantages of oil cleanser with high cleaning properties while simultaneous improving its permeability and moisture capacity, providing restoring elasticity and regenerative capacity of the skin without leaving an unpleasant oil film on the skin surface.
7 cl, 11 ex, 1 tbl
SUBSTANCE: invention relates to field of food industry, biotechnology and deals with antibacterial composition and strain of bacteriophage Escherichia coli, used for obtaining said composition. Characterised composition includes filtrate of Escherichia coli phage lysate, obtained with application of strain of bacteriophage Escherichia coli, deposited in collection of museum of microorganisms of Federal Budget Institution of Science "State Research Centre for Applied Microbiology and Biotechnology" of the Federal Service on Customers' Rights Protection and Human Well-being Surveillance (FBIS SRC AMB of Rospotrebnadzor) under number Ph 64, filtrate of Escherichia coli phage lysate, containing coli bacteriophage, filtrate of staphylococcus phage lysate, filtrate of salmonella phage lysate, filtrate of Listeria monocyctogenes phage lysate and target additives in amount 1.0÷95.0 wt % of composition weight.
EFFECT: claimed composition has required spectrum of specific activity due to inclusion of polyvalent bacteriophage and can be used in production of biologically active additives and food additives.
11 cl, 1 tbl, 13 ex