RU2262350C2 - Vaccine for prophylaxis and immunotherapy of human and animal diseases caused by pathogenic and opportunistic gram-negative microorganisms of intestine group and their exotoxins and method for its preparing (variants), immunoglobulin preparation (variant) and method for its preparing, immunobiological preparation polycomponent vaccine - Google Patents

Abstract

FIELD: biotechnology, microbiology, medicine, veterinary science.

SUBSTANCE: for preparing vaccine toxigenic strains of S. dysenteriae R-forms are grown, cells are subjected for lysis by treatment with chloroform, mixture is centrifuged and prepared supernatant is treated with saturated monobasic carboxylic acid or their derivatives and pH value is brought about to 3.0-5.0. Mixture if centrifuged and precipitate containing corpuscular antigens and shigellosis exotoxin are obtained. Precipitate is dissolved in buffer and pH value is brought about to 7.5-9.0. Then formalin is added in the amount 0.4-0.8% of the solution volume, or benzoic acid or benzoic acid salts are added in the amount 0.07-4.0% of the solution volume, or a mixture consisting of formalin and benzoic acid or benzoic acid salt solutions in the amount 0.10.3% and 0.03-2.5% of the solution volume, respectively. The solution is kept at temperature 30-60°C for 2 h - 60 days to provide the conversion of exotoxin to anatoxin and vaccine is prepared. Another variant of the claimed invention involves additional treatment with formalin or benzoic acid or benzoic acid salts to provide conversion of exotoxin to anatoxin, vaccine is prepared followed by its bagging and corking. For preparing immunoglobulin preparation animals are immunized with vaccine prepared by abovementioned methods followed by taking off blood, milk and/or colostrums, and/or blood, immunoglobulin fraction is prepared, sterilized, bagged and corked. This preparation is a component of the immunobiological preparation. The immunobiological preparation comprises the immunoglobulin preparation and at least one component taken among the following row: human and/or animal immunoglobulin preparations, lactoferrin, enzymes, inhibitors of proteolytic enzymes, human and/or animal normoflora preparations, yeast, vitamins, vitamin-like substances, human and/or animal proteins of acute phase, human and/or animal cytokines, higher plants components, lower plants components, components of natural origin products, apiculture products, enterosorbents, antibiotics, antibacterial chemopreparations, sulfanilamide drugs, antibacterial, anti-tuberculosis, antiviral preparations, antifungal antibiotics, synthetic antifungal preparations, stimulators of metabolic processes, antioxidants, mineral supplements, carbohydrates, lipids, replaceable and/or essential amino acids, organic acids, alkaloids, glycosides, taste supplements, aromatic supplements, base for suppositories, base for ointment formulations, technological additives for tableting, or their mixture. Invention provides preparing preparations eliciting antigenic activity with respect to broad species of pathogenic and opportunistic gram-negative microorganisms of intestine group and their exotoxins and therefore eliciting with prophylactic and curative effect with respect to diseases causing with these microorganisms.

EFFECT: improved preparing method, valuable properties of vaccine.

13 cl, 112 ex

Description

The group of inventions relates to biotechnology, medicine, veterinary medicine and can be widely used in the production of vaccines against pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group, immunoglobulin and immunobiological preparations, in treatment, in prevention, as well as in the manufacture of food products, animal feed and dietary supplements.

It is known that the pathogenesis of diarrhea accompanying the course of coli infection, dysentery, salmonellosis, and diarrhea caused by protein and other gram-negative microorganisms of the intestinal group, is based on the action of exotoxins (neurotoxins), which have enterotoxic activity and possess immunogenic antigenicity, that are pathogenic between themselves and with cholerogen. Differences in the pathogenetic manifestation of these diseases largely depend on the level of exotoxin release by these microorganisms (Gabidullin Z.G., Zhukova S.L., Jezepchuk Yu.V., Bondarenko V.M. // ZhMEI. - 1989. - N12. - p. 14-16; Dzhavets E., Melnik J.L., Eidelberg E.A. Guidelines for Medical Microbiology.Lane from English.- M .: Medicine, 1982. - v.2. - 384 p .; Ivanov NR, Ermolov VI Immune preparations of milk and their use in acute intestinal infections in children - Saratov: Publishing house of the Saratov University, 1985. - 120 p .; Kamzolkina NB, Karavaeva L.T. , Japaridze M.N. Profile the tics of especially dangerous infections. - M., 1977. - p. 51; Korotaev A.I., Babichev S.A. Medical Microbiology, Immunology and Virology.- St. Petersburg: Special Literature. - 1998. - 592 p. ; Medical Microbiology. / Under the editorship of A.M. Korolyuk and V. B. Sboychakov. - St. Petersburg. - 1999. - Part 1. - 272 p .; Fedorov Yu.N. Brylin A.P. , Dostaleva M.I. // Agricultural Biology. - 1988. - N5. - pp. 117-124; Acres S. //J.Dairy Sci. - 1985. - vol. 68. - N1. p.229-256; Barber C., Eyian E., Keyelar Y. // Rev. Immunol. - 1969. - vol. 33. - p.101; Climents J., Finkelstein R. // Infect. Immun. - 1978. - vol. 22. - N3. p.709-713; Mulezyh M., Adamus G., Witnowska D., Romanowska E. //Arch.Immunol.Ther.Exp. - 1981. - vol. 29. - p.85; Mulles W., Schlecht S., Westphal O. // Zbl. Bakt.Hyg.I.Abt.Orig.A. - 1980 .-- vol. 64. - p.248; O, Brien A.D., Chen M.E., Holmes R.K. ct al. // Lancet. - 1984. - vol. 14. p.77-78 Supotnitsky M.V. - Microorganisms, toxins and epidemics. - M.: University book. - 2000. - 376 p.).

It has been experimentally proved that immunity against enteropathogenic Escherichia coli, Shigella and other gram-negative microorganisms is largely determined by the presence of antitoxic antibodies, primarily to the thermolabile component of exotoxin (Vartanyan Yu.P., Severtseva MK, Stanislavsky E.S.// ZHMEI. - 1980. - N8. - p. 87-90; Emody L., Ketyi I., Kuch B., Pacsa S. // Acta Sci Hung. - 1979. - vol.26. - N3. - p. 223-241; Klipstein F., Engert F., Sort H. // Infect. Immun. - 1980. - vol. 28. - N1. - p.163-170). The severity of antitoxic immunity depends on many reasons, in particular on the immunogenicity of toxoids. For example, Shigella exotoxins of different species are immunogenetically related, but only S.dysenteriae 1 exotoxin has the most powerful cytotoxic properties, and the toxoid obtained from it has pronounced immunogenicity. On the one hand, this is due to the fact that the exotoxin of the species S.dysenteriae (Shiga toxin is an old, but still used name) has a higher molecular weight, on the other hand, the exotoxins produced by S. Sonnei and S. Flexneri are produced in 1000 or more times less. Therefore, both cross-reacting and specific antibodies are more pronounced after vaccination of animals with Shiga-toxoid (Supotnitsky M.V. Microorganisms, toxins and epidemics. - M.: University book. - 2000. - 376 p .; Golubeva IV, Kilesso V.A., Kiseleva B.S. et al. Enterobacteria: a guide for doctors. - M .: Medicine, 1985. - 321 p .; Kamzolkina NB, Epstein-Litvak R.V., Kokorina T. A. // BEBIM., - 1971. - N2. - p. 70; Timakov V.D., Petrovskaya V.G., Bondarenko V.M. Biological and genetic characteristics of bacteria of the genus Shigella. - M .: Medicine. - 1980 .-- 296 p .; Keusch GT, lacewicz M. // J.Infect. Dis. - 197 7. - vol. 135. - p. 511; O, Brien A.D., Thompson M.K., Gemski P., Doctor B.P. et al. // Infect.Immun. - 1971. - vol.15. - p.796).

A comparative immunochemical analysis of the corpuscular antigens of gram-negative bacteria of the intestinal group, especially enterobacteria, also shows a significant antigenic relationship of these microorganisms. The formation of chromosomal hybrids between E. Coli and bacteria of the genera Salmonella and Shigella indicates a very high degree of genetic homology of these bacteria, which confirms the comparison of chromosomal maps. Of particular note is the kinship between E. Coli and Shigella. The genomes of these microorganisms are related by 85-90%. The high degree of similarity of the genomes also determines the high similarity of the antigenic properties of the Shigella and E. Coli strains, which is expressed in the presence of cross-reactions with diagnostic agglutinating sera. It is important to note that the antigenic composition of most representatives of pathogenic microorganisms of the intestinal group of humans and animals is approximately the same (Golubeva I.V., Kilesso V.A., Kiseleva B.S. et al. - Enterobacteria: a guide for doctors. - M .: Medicine , 1985. - 321 p .; Korotaev AM, Babichev SA Medical Microbiology, Immunology and Virology. - St. Petersburg: Special Literature. - 1998. - 592 p .; Medical Microbiology. // Ed. A. M. Korolyuk and V. B. Sboychakov. - St. Petersburg. - 1999. - Part 1. - 272 p .; Medical Microbiology. // Edited by V.I. Pokrovsky and O.K. Poz Deeva. - M.: GEOTAR Medicine, 1999. - 1200 p .; Mikhailov N.V. // Mater. International Symposium dedicated to the year of L. Pasteur. - St. Petersburg, 1995. - p.66; Steinier R., Edelberg E., Ingram J. The World of Microbes (Translated from English) - Moscow: Mir, 1979. - v. 3. - 486 p.).

A major problem for medicine is zoonoses - infections common to humans and animals (Makarov V.V., Vorobyov A.A. // ZhMEI. - 1999. - N4. - p.111-115). Up to 70% of all diarrheal diseases in children under 5 years of age have a nutritional etiology. For example, the incidence of salmonellosis in some European countries reaches 300 cases per 100 thousand people with a mortality rate of 3%. In recent years, in many developed countries (USA, Canada, Great Britain, Japan, etc.), colibacteriosis of animals has been under the scrutiny of veterinary and medical specialists, as well as WHO, since the Escherichia producing Shiga-like began to play an important role in human infectious pathology toxin or verotoxin (VTEC), in particular the serovar E. Coli 0157: H7 (ECO). In Russia, the situation is even more acute (Golubeva I.V., Kilesso V.A., Kiseleva B.S. et al. - Enterobacteria: a guide for doctors. - M .: Medicine, 1985. - 321 p .; Golutva N .K., Hazenson LB, Safonova N.V. // Mater. International Sympos. Dedicated to the year of Pasteur. - St. Petersburg. - 1995. - p. 53; Kulikovsky A.V., Panin A .N., Sosnina V.V. // Veterinary medicine. - 1997. - N3. - p.25-27; Makarov V.V., Vorobev A.A. // ZhMEI. - 1999. - N4. - p. 111-115; Medical Microbiology. // Under the editorship of V.I. Pokrovsky and O.K. Pozdeev. - M.: GEOTAR Medicine, 1999. - 1200 p.; Sergeevnin V.I. // Epidemiology and Infectious Diseases. - 1974.- N4. - p. 32-34; Steinier R., Edelberg E., Ingram J. The World of Microbes. Translated from English - M .: Mir, 1979. - v. 3. - 486 p .; WHO Consultation on Selected Emerging Foodbome Dis. // WHO / CDS / VPH. - 1995; Zoonoses. // WHO Veterinary Public Health Unit. Publications and Documents. - 1995).

As follows from the analysis, there are currently no domestic and imported vaccines containing shigellosis toxoids, in addition, the domestic industry does not produce shigella vaccines (Medunitsyn N.V. Vaccinology. - M., Triada - X, 1999, 272 p. .).

In earlier sources, a dysenteric vaccine from S. Flexneri or S. Sonne and a divacin from them are mentioned (Guidance on the vaccine and serum case. - // Under the editorship of PN Burgasov. M .: "Medicine", 1978, 440 pp. .). However, these vaccines did not find application in practical medicine, as due to the lack of toxoid in them after vaccination, people do not produce antitoxic antibodies, in addition, there is a low immunogenicity of the drugs.

Thus, the vaccine and the method for its preparation, which would contain shigellosis toxoid, as well as toxoids used to treat and prevent a wide range of diseases caused by opportunistic and pathogenic gram-negative microorganisms of the intestinal group, are not known from the prior art.

In veterinary medicine, a multispecific hyperimmune serum has been developed against rotavirus, corona, herpes viruses and E. Coli (K 99, A 20) for local protection and immunotherapy of mixed forms of diarrhea of newborn calves (Patent RU 2043772, 09/20/95; Patent RU 2054289, 20.02. 96; Patent RU 2086236, 08/10/97). This drug does not have antitoxic activity and does not contain antibodies to shigella.

In the United States a patented method for producing an immune preparation obtained from milk and colostrum of immune cows and possessing activity against enterobacteria, pathogenic for humans and farm animals (US Patent N4402938, class A 61 K 39/00, 1983; German application N3924420, class A 61 K 35/20, 1991).

In our country, two oral immunoglobulin preparations obtained from cow immune colostrum are used in medical practice - the so-called immune lactoglobulins (a name that has become established in Russia): against opportunistic bacteria and salmonella (Medunitsyn N.V. Vaccinology. - M., "Triad-X", 1999, 272 pp .; Soboleva S.V. // Abstract of dissertation in the form of a scientific report ... Doctor of Medical Science - Rostov-on-Don, 1991. - 38 pp.) and against Escherichia coli and Proteus (Medunitsyn N.V. Vaccinology. - M., "Triad-X", 1999, 272 pp .; Soboleva S.V. // Abstract of thesis in the form of a scientific report yes ... doctor of medical science. - Rostov-on-Don, 1991. - 38 p .; Soboleva S.V., Ermolov V.I., Valentsev V.E. et al. // Sat. questions of immunology, allergology and molecular biology ". - Krasnodar, 1987. - P. 63. - p.119-121; Soboleva S.V., Mineeva L.D., Rabinovich V.D., Fedorova T.A. / / J. Microbiol. - 1989. - N8. - p. 7-12).

As shown by the analysis of the prior art in relation to immunoglobulin preparations, there is no information in the literature about immunoglobulin preparations containing antibodies to any toxins of pathogenic gram-negative microorganisms of the intestinal group and shigella. All known immune immunoglobulin preparations contain antibodies only to the cell wall of microorganisms.

The closest technical solution in relation to the immunoglobulin preparation and the method for its preparation will be the anti-lipoprotein lactoglobulin, which is an immunoglobulin preparation obtained by immunizing cows, collecting colostrum, obtaining immune serum (Ivanov N.R., Ermolov V.I. Immune milk preparations and their use in acute intestinal infections in children (Saratov, Publishing House, Sarat., 1985, p. 35-44). The disadvantage of this immunoglobulin preparation is the low specific activity, the absence of antitoxic antibodies and antibodies to shigella, which is confirmed experimentally in living models.

The objective of the claimed invention is the creation and preparation of a highly effective and low toxic vaccine for the immunoprophylaxis and immunotherapy of diseases of humans and animals caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins, the creation of immunoglobulin preparations with therapeutic and prophylactic effects against diseases of humans and animals, caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their ek zotoxins, the creation of immunobiological preparations with therapeutic and prophylactic effects against a wide range of diseases of the internal and external organs of humans and animals caused by various pathogenic and conditionally pathogenic gram-negative and gram-positive microorganisms and their toxins, viruses, fungi, protozoa, helminths.

This object is achieved in that in order to obtain a vaccine for the immunoprophylaxis and immunotherapy of diseases of humans and animals caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins, toxigenic strains of the R-form S.dysenteriae are grown, the cells are lysed by treatment with chloroform, centrifuged then the resulting supernatant is treated with saturated monobasic carboxylic acids and their derivatives, for example trichloroacetic acid, caprylic acid to pH 3.0-5.0, centrifuged to obtain a precipitate containing corpuscular antigens and shigellous exotoxin, dissolve the precipitate in a buffer to obtain a solution of pH 7.5-9.0, add formalin in an amount of 0.4-0.8% of the amount of solution, or benzoic acid, or salt of benzoic acid in an amount of 0.07-4.0% of the amount of solution, or a mixture thereof, while the amount of formalin is 0.1-0.3% of the amount of solution, the amount of benzoic acid or its salts 0.03-2.5% of the amount of the solution, withstand the solution at a temperature of 30-60 ° C for 2 hours - 60 days to the transition of exotoxin to toxoid to obtain a vaccine. Before packaging, the vaccine can be freeze dried to obtain granules and powders. Before packaging, an ointment base or suppository base can be added to the vaccine.

Another objective is achieved by the fact that to obtain a vaccine for immunoprophylaxis and immunotherapy of diseases of humans and animals caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins, toxigenic strains of the R form of S.dysenteriae are grown, cell lysis is performed by treatment with chloroform , centrifuged, then the resulting supernatant is treated with saturated monobasic carboxylic acids and their derivatives, for example trichloroacetic acid, capri hydrochloric acid, to a pH of 3.0-5.0, centrifuged to obtain a precipitate containing corpuscular antigens and shigellosis exotoxin, dissolve the precipitate in a buffer to obtain a solution of pH 7.5-9.0, add formalin in an amount of 0.4-0 , 8% of the amount of solution, or benzoic acid, or salts of benzoic acid in an amount of 0.07-4.0% of the amount of solution, or a mixture thereof, while the amount of formalin leaves 0.1-0.3% of the amount of solution, the amount of benzoic acid or its salts 0.03-2.5% of the amount of the solution, withstand the solution at a temperature of 30-60 ° C for 2 hours — 60 days, then additional processing is carried out by adjusting the amount of free formaldehyde to 0.18-0.25% of the amount of solution, or free benzoic acid, or its salts in an amount of 0.07-4.0% of the amount of solution, or their mixture, while the amount of free formaldehyde is 0.037-0.12% of the amount of solution, and the amount of free benzoic acid or its salts 0.03-2.5% of the amount of solution is kept at 30-60 ° C for 2 hours - 30 days before the transition of exotoxin to toxoid to obtain a vaccine, then carry out bulk ku and capping. In this case, before packaging, the final product can be subjected to freeze drying to obtain granules or powders.

Before packaging, an ointment base or suppository base can be added to the vaccine.

The vaccine for immunoprophylaxis and immunotherapy of diseases of humans and animals caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins, when growing S.dyscnteriae on casein broth, is subjected to trichloroacetic acid treatment of the supernatant, the vaccine obtained by the above methods is an antigens and shinuella fraction contains protein in an amount of 10-15 mg / ml, nucleic acids 0.2-0.5 mg / ml, lipids - traces. When a microbial culture is grown on other nutrient media, the composition of the vaccine, as shown by experiments, can vary in composition.

This vaccine can be used for immunoprophylaxis and immunotherapy of diseases of humans and animals caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins.

To obtain an immunoglobulin preparation, animals are immunized with a vaccine for the immunoprophylaxis of diseases in humans and animals caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins obtained by the above methods, and then the animals are collected immune milk and / or colostrum and / or blood receive immunoglobulin fraction. After sterilization, drying can be carried out.

The immunoglobulin preparation obtained by the above method contains a fraction of immunoglobulins G, A and M in the ratio (10-50) :( 0.01-5) :( 1-10). As studies have shown, the ratio of immunoglobulins in a preparation is completely dependent on the raw materials used and can vary greatly depending on the breed of animals, their age, lactation time, season, diet, nutritional composition, past illnesses, health conditions, genetic anomalies, sometimes found in ruminants of congenital natural IgA immunodeficiency.

An immunoglobulin preparation may additionally contain at least one component selected from the range: flavoring agents, aromatic additives, suppository base, ointment base, and tabletting processing aids.

An immunobiological preparation contains an immunoglobulin preparation and at least one component selected from the series: human and / or animal immunoglobulin preparations, lactoferrin, enzymes, proteolytic enzyme inhibitors, human and / or animal normoflora preparations, yeast, vitamins, vitamin-like substances, acute phase proteins human and / or animal, human and / or animal cytokines, components of higher plants, components of lower plants, components of products of natural origin, beekeeping products, enteroso bents, antibiotics, antimicrobial chemotherapeutic agents, sulfa drugs, antimicrobial and antiparasitic drugs of natural origin, antituberculosis drugs, antiviral drugs, antifungal antibiotics, synthetic antifungal agents, metabolic stimulants, antioxidants, mineral additives, carbohydrates, lipids, essential or organic acids, alkaloids, glycosides, flavorings, aromatic additives, suppository base, again for ointment form, processing aids for tabletting or a mixture thereof.

In this case, human and / or animal immunoglobulin preparations are selected from the series: immunoglobulin fraction, immunoglobulin G, immunoglobulin A, immunoglobulin M, immunoglobulin T, fragments of immunoglobulins.

Enzymes are selected from the following series: lactoperoxidase, lysozyme, trypsin, chymotrypsin, chymopsin, terrilithin, terridekase, ribonuclease, deoxyribunclease, collagenase, asperase, lysoamidase, profesim, caripazin, pepsin, natural gastric juice, pepsimra, pepsimra solomon, pepsidra solomon, pepsilazin , nygedase, panzinom forte, pancurmen, festal, digestal, enzistal, mesim forte, lidase, ronidase, cytochrome C, penicillinase.

Proteolysis inhibitors are selected from the range: pantripine, ingitrile, contracal, gordox.

Normoflora preparations of humans and / or animals are selected from the series: bifidobacteria, lactobacilli, streptococci, strains of apatogenic Escherichia coli, saccharomycetes.

Yeast selected from a number: brewer's yeast, baker's yeast.

Vitamins are selected from the series: A, group B, D, K, P, PP, U, E, calcium pantothenate, panthenol.

Vitamin-like substances are selected from the series: methyl methionine sulfonium chloride, calcium pangamate, choline chloride, lipoic acid, lipamide, orotic acid, pangamic acid, para-aminobenzoic acid, inositol, carnitine.

Proteins of the acute phase of humans and / or animals are selected from the series: haptoglobin, α-fetoprotein, orozomcoid, ceruloplasmin, C-reactive protein, α1-antitrypsin, α1-antichymotrypsin, Gc-globulin.

Cytokines are selected from the series: interferons (IF), interleukins (IL), tumor necrosis factors (TNF), transforming growth factor β (TGFβ), colony stimulating factors (CSF), leukemia suppression factor (LIF), CD95.

The fractions, which are infusions and / or extracts, and / or decoctions, and / or extracts, and / or powder of higher plants, are selected from the series: aloe, medicinal marshmallow, orange, Aralia manchurian, common apricot, American agave, Indian adenosma, calamus marsh, aromatic ayovan, prickly acantopanax, white acacia, Alpinia officinalis, spikelet amaranth, real pineapple, andrographis paniculata, asphodelian anamarrena, Chinese angelica, common anise, edible peanuts, blackberry, cannabis, lamb prickly artichoke, woolly astragalus, ovate atractylodes, marsh rosemary, common badass, true star anise, common basil, cultivated banana, Amur velvet, ram, common barberry, hanging birch, immortelle, boletus rosacea, rosemary roselacea, rosemacea rosacea rosacea , lingonberry, ivy buds, elderberry black, initial letter, medicinal borage, Valerian officinalis, vanilla, blue cornflower, three-leafed shift, common loosestrife, medicinal verbena, Veronica officinalis, cultivated grapes, sleeping pills, cherries, sickle beetle, curly thistle, Medicinal teasel, elm rusty, gardenia jasmiformes, open harpophytum open, fragrant cloves, crowned hedgehog crowned, meadow geranium, blue-headed gibiscus, Syrian syrup, highlander bird, gentian gentian, sowing peas, panicle hydrangea, adonis, goryanka, pomegranate, grapefruit, sowing buckwheat, common pear, guarana kupana, heart-shaped guttuinia, de vyaslil high, Chinese dereza, fluffy dioscorea, medicinal sweet clover, common oak, oregano, officinalis officinalis, blueberry blackberry, Canadian yellow root, ginseng, Altai honeysuckle, tall zantoxilum, pepper zanthoxylum, middle asterisk, St. John's wort, wild strawberry, wild strawberry, wild strawberry, wild strawberry, forest strawberry common, Centaury common, scented bison, white willow, medicinal ginger, emperor cylindrical, common fig, hyssop officinalis, Seneg’s source, yohimbe, real cocoa, calendula vein, common viburnum, capers prickly, white cabbage, cardamom real, potato, cassia holly, horse chestnut, medicinal dogwood, meadow clover, seedlings, black cohosh daursky, strawberries, marsh cranberries, brilliant cola, condurango, coriander seeds, coriander seed, real cinnamon, catnip, coffee tree, dioecious nettle, medicinal blood meal, buckthorn laxative, ordinary corn, turmeric long, meadowsweet, spiky lavender, noble laurel, cinquefoil and goose, spreading quinoa, safflower-like levzea, sowing flax, capillary lesdesa, capillary lesdesa, common hazel, lemon, Chinese schisandra, chilebeaceae linden, hearty linden, large burdock, walnut-bearing lotus, bulbous luciferous, luciferous medicinal poppy samoseyka, raspberry ordinary, mallow forest, mango syzyo, Chinese mandarin, European olive, mother-and-machiha ordinary, mate, madder dye, lemon balm medicinal, miroballan, myrrh tree, juniper ordinary venous, Momordika Kokhinkhinskaya, Morinda officinalis, sowing carrots, seaweed, peppermint, buckthorn, oat, dandelion, walnut, nutmeg, digitalis, hay fenugreek, papaya, passiflora male-red meat bag, pasta allspice, water pepper, cava pepper, chilli pepper, cubebe pepper, black pepper, Javanese pepper, common peach, garden parsley, tansy ordinary, trigeminal pinella, dodging peony, Siberian fir, Indian plumeha, Japanese ika, large plantain, common sunflower, bitter wormwood, bitter oranges, kidney tea, hazel leaf psoralea, parsnip, lobed pueraria, heartwort, pfaffia, soft wheat, creeping wheatgrass, spotted milk thistle, rhubarb, radish , planting rice, pink Rhodiola, rye, medicinal rosemary, pharmacy chamomile, sundew, round rowan, mountain ash, Chinese sassaparil, common beets, celery odorous, senna, creeping selena, homemade sida, black dyna, red currant, golden currant, common daisy, licorice naked, malt, hill hodgepodge, burdock saussure, Japanese sophora, cultivated soybean, asparagus officinalis, plowed stalnik, cornflower stem, stevia, tuberous stamen, dried stalk, gingerbread , tanning sum, Indian tamarind, prickly tartar, thorns, thermopsis, bearberry ordinary, Jerusalem artichoke, black poplar, mulberry tree, ordinary caraway seeds, pumpkin, yarrow, garden dill, unabi, unkari hair thawed, ordinary haricot, fennel, common physalis, hanging forsythia, field horsetail, ordinary hop, horseradish, Indian chrysanthemum, Asian centella, chicory ordinary, thyme ordinary, plantain, three-part sequence, bird cherry, common blueberry, garlic, Chistyk is alphabetical, salvia officinalis, common chandra, saffron, rosehip cinnamon, Scutellaria baicalens, garden spinach, curly sorrel, eucalyptus, Eucommia beetle, prickly Eleutherococcus, elsholzi patrena, embliki officinalis, Echinacea purpurea, yucca, apple tree, European ash, barley.

The fractions, which are infusions, and / or extracts, and / or decoctions, and / or extracts, and / or powder of lower plants, are selected from the series: porcini mushroom, varnished fungus mushroom, real mushroom mushroom, tea comb (tea mushroom), sugar kelp (seaweed), matake, pakhima, reishi, spirulina, bubbly fucus, chlorella, Icelandic cetraria, black birch mushroom - chaga, shiitake.

The fractions, which are infusions, and / or extracts, and / or decoctions, and / or extracts, and / or powder of products of natural origin, are selected from the series: shark cartilage, bezoar stone, beeswax, farm animals muscle powder, gelatin, bile , squid, bone tissue, crabs, shrimp, animal blood, mussels, mollusks, condensed colostrum, salmon milk, skimmed milk powder, condensed milk, whey, sea urchins, ants, pantogematogen, antlers, pantocrine, bear and deer genitals, mollusk shells, fish powder, internal organs of pike, scolopendra, scorpion, chitin, cartilage of farm animals, earthworms, tortoiseshell, black-tailed snake, egg. Beekeeping products are selected from the range: propolis, honey, royal jelly (apilak), plant pollen.

Enterosorbents are selected from the range: activated carbon preparations, SKN enterosorbent, carbolong, polyphepan, zeolites, gummivit, kaolin, kieselguhr, montmorillonite.

Antibiotics are selected from the series: penicillin group preparations, cephalosporin groups, cefamycins, carbapenems, monobactams, tetracyclines, aminoglycoside antibiotics, macrolides antibiotics, azalides, lincomycin group preparations, chloramphenicol, ristamycin polyginsin, fusidin spectinomycin, preparations of the streptomycin group, rifampicin group, cycloserine group, florimycin group, rifabutin, nystatin, levorin, amphotericin, amphoglucamine, mycoheptin, primycin.

Antimicrobial chemotherapeutic agents are selected from the series: 8-hydroxyquinoline derivatives, naphthyridine derivatives, quinolones, fluoroquinolones, quinoxaline derivatives, nitrofuran derivatives.

Sulfanilamide preparations are selected from the following series: streptocide, norsulfazole, sulfazine, sulfargin, sulfadimezin, etazole, sulfacyl, urosulfan, sulfapyridazine, sulfamonomethoxin, sulfadimethoxine, sulfalene, sulfalene-meglumine, cotrimoxazole, algimaf, sulfazalazalazalazalazalazalazalazalazalazalazalazalazalazalazalazalazalazalazalazal

Antimicrobial and antiparasitic drugs of natural origin are selected from the series: sodium usnin, novimanin, sanguirytrin, umcalor, eucalymin, chlorophyllipt, arsnarin, baliz-2, ekteritsid, tomitsid, marigold flowers, tincture of Japanese sophora, allilchap.

Antituberculosis drugs are selected from the series: isonicotinic acid hydrazide, its derivatives and analogues, para-aminosalicylic acid derivatives, preparations of the ethambutol group, pyrazinamide group, thioacetazone group, solutizone group.

Antiviral drugs of various groups are selected from the following series: intferon, induferon inducers, acyclovir, ganciclovir, ribamidine, zidovudine, idoxuridine, lamivudine, didanosine, valacyclovir, famciclovir, ribavirin, indinavirin, dendinfinidin, remindinidin, remantina, remantina, remantina, remantina, remantine , tebrofen, riodoxol, frorenal, flacoside, alpizarin, helepin, megosin, gossypol.

Antifungal antibiotics selected from the series: griseofulvin, nystatin, levorin, levorin sodium salt, amphotericin B, amphoglucamine, mycoheptin.

Synthetic antifungal drugs are selected from the series: clotrimazole, ketokenazole, econazole, miconazole, mycozolone, isoconazole, fluconazole, bifonazole, amorolfine, itraconazole, terbinafine, naphthifin, decamine, mycoseptin, hinofinfin, nitrofinfin, nitrofinfin, nitrofinfin, nitrofinfin, nitrofinfin, nitrofinfin, nitrofinfin, nitrofinfin, nitrofinfin, nitrofin, nitrofin, nitrofin, nitrofin, nitrofin, nitrofin, nitrofin, nitrofen.

Drugs that stimulate metabolic processes are selected from the series: methyluracil, pentoxyl, potassium orotate, leucogen, adenosine triphosphoric acid, phosphaden, etaden, phosphocreatine, riboxin, dalargin, semax, dipromonium, carnitine chloride, mildronate, atsemin, polyethylene oxide, polyethylene oxide, catalytic, , sargramostim, molgramostim, lenograstim.

Antioxidants are selected from the range: vitamins A and E, dibunol, superoxide dismutase, emoxipin, mexidol, ubinone.

Mineral additives are selected from the series: calcium, potassium, sodium, iron, zinc, copper, manganese, cobalt, chromium, molybdenum, phosphorus, sulfur, magnesium, chlorine, fluorine, silicon, bromine, silver, boron, vanadium, germanium preparations.

Carbohydrates are selected from the series: glucose, fructose, lactose, sucrose, maltose, inulin, starch, dietary fiber, pectin, gums, mucus.

Lipids are selected from the range: saturated saturated and unsaturated acids, almond oil, peach oil, sea buckthorn oil, rosehip oil, castor oil, corn oil, cottonseed oil, peanut oil, bran food oil, sunflower oil, pumpkin seed oil, phospholipids, sterols, waxes.

Replaceable and / or irreplaceable amino acids are selected from the range: arginine, alanine, cysteine, cystine, glutamic acid, glutamine, glycine, histidine, leucine, lysine, methionine, ornithine, proline, phenylalanine, tyrosine, valine.

Organic acids are selected from the series: malic, citric, hydroxycitric, tartronic, tartaric, oxalic, salicylic, formic, acetic, benzoic, hydroxybenzoic, lipoic, protocatechonic, pyrocatechonic, gallic, succinic, malonic, hydroxyboric, nicotine.

Alkaloids are selected from the series: caffeine, salsolin, berberine, palmatine, tyroricin, columbamine, capsaicin, viburnin, amyrin, theobromine, tsofillin, xanthine, silymarin, stahydrin.

Glycosides are selected from the series: steroidal saponins, triterpene saponins, antraglycosides, bitter glycosides (iridoids), cardiac glycosides.

The immunobiological preparation can be a lyophilized powder, granules, capsules.

The immunobiological preparation may additionally contain pharmaceutical components for tabletting.

The immunobiological preparation may additionally contain pharmaceutical components for the preparation of suppositories.

The immunobiological preparation may additionally contain pharmaceutical components for the preparation of ointment forms.

An immunoglobulin preparation can be used for the prevention and treatment of human and animal diseases caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins.

An immunobiological preparation can be used to treat and prevent a wide range of diseases of internal and external organs of humans and animals caused by various pathogenic and conditionally pathogenic gram-negative and gram-positive microorganisms and their toxins, viruses, fungi, protozoa, and helminths.

A multicomponent vaccine may contain at least two components, while one of the components contains a vaccine for immunoprophylaxis and immunotherapy of diseases of humans and animals caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins obtained by the above methods.

Evidence of the effectiveness of the vaccine for the prevention of diseases of humans and animals caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins against pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins are laboratory tests on animals, cell cultures and hemagglutination reactions as the vaccine itself and immunoglobulin preparations obtained from colostrum, milk and blood immunized with this vaccine domestic and agricultural animals, as well as clinical trials of an anti-shigellosis immune immunoglobulin preparation (anti-shigellosis lactoglobulin) obtained from colostrum of immune cows.

When immunized with a vaccine for the immunoprophylaxis and immunotherapy of diseases of humans and animals caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins, we have obtained antibodies to a wide range of conditionally pathogenic and pathogenic gram-negative microorganisms of the intestinal group and their exotoxins.

Experiments have been carried out in comparison with checking the antitoxic and antibacterial protection of anti-Shigella drugs (immunoglobulin preparations of goat and cow), lactoglobulins (preparations of immune immunoglobulin preparations) of anti-lipoprotein and against opportunistic bacteria Salmonella (at the request of GISK named after L.A. Tarasovich), , as well as comparing anti-shigellosis drugs with normal human gamma globulin.

Experiments on the neutralization of Shigella exotoxin with anti-Shigella drugs and immunoglobulin G isolated from it were performed on mice and HeLa cell culture. When setting on mice, the intraperitoneal method and the test of swelling of the mouse paws were used.

At 2.4-2.5 LD _50, immune anti-shigellosis drugs on average 344 times higher in antitoxic activity than non-immune milk preparations, and at 3.5-4.4 LD ₅₀ - on average 578 times. Comparing the IgG isolated from the immune and non-immune preparation of goat milk, we observe a difference in antitoxic activity at 2.5-3.1 LD ₅₀ 520 times, at 3.9-4.2 LD ₅₀ 627 times. Anti-shigellosis preparations exceeded the anti-lipoprotein lactoglobulin by 50-400 times in antitoxic activity, the difference between the first and lactoglobulin against opportunistic bacteria and salmonella was 166-367 times or more. Anti-Shigella drugs exceeded the activity of non-immune lactoglobulin 140-366 times. If we take the level of antitoxic activity of a non-immune preparation as zero, then we can say that lactoglobulin against opportunistic bacteria and Salmonella does not have antitoxic activity, and anti-lipoprotein also does not have these antibodies or contains very few of them.

In the test of mouse paw edema, an anti-shigellosis drug completely prevented the development of paw edema at a dose of 0.093-0.187 mg, and IgG isolated from it - 0.062-0.125 mg, while the administration of an non-immune preparation and obtained IgG at a dose of 1 mg showed a complete absence of a positive effect , i.e. swelling of the legs persisted. It seemed impossible to experiment with the introduction into the paw of a mouse weighing 10 g more than 1 mg of non-immune preparations.

When checking the antitoxic activity of the anti-shigellosis immunoglobulin on HeLa cells, it was determined that for the neutralization of CTD ₁₀₀ , 128-198 times less immune preparation than non-immune is required. Moreover, when compared with the antitoxic intraperitoneal test in mice, the cytotoxic test on HeLa cells gives more uniform results from experiment to experiment.

The antibacterial protective activity of anti-shigellosis drugs was tested in mice using the intraperitoneal method with respect to S.sonnei, S. flexneri 2a, S.dyscnteriae 1, S.typhi and Ps.aeruginosa.

For preparations obtained from colostrum of immune and non-immune goats, the difference in protective activity upon infection with 3-4 LD ₅₀ of S.sonnei culture averaged 71.4 times. The protective activity index of coliprotein lactoglobulins against opportunistic bacteria and salmonella was lower at 13.6–20.0 compared to the anti-shigellosis drug. When mice were infected with S.dyscnteriae 1, the difference in protective activity between anti-shigellosis and non-immune preparations averaged 96.2 times, between anti-shigellosis and anti-lipoprotein - 96 times, anti-shigellosis and lactoglobulin against opportunistic bacteria and salmonella - more than 76 times.

Protivoshigelleznye immunoglobulin preparations according sredneeffektivnym protective dose against S.flexneri 2a in mice nonimmune superior drug with 3,2 LD ₅₀ in 27-66 times, 4,2-5,6 LD ₅₀ - 62-75 times with 11 , 5-19.0 LD ₅₀ - 73.8-289 times, the activity of anti-schigellosis drugs is higher than the anti-lipoprotein one at 3.2-5.6 LD _{50 by} 17.7-36 times, at 11.5-13.0 LD ₅₀ - 94-108 times, between the anti-shigellosis lactoglobulin preparations and normal gamma globulin, the difference at 3.2-5.6 LD ₅₀ is 27.6-40 times, the activity of the anti-lipoprotein drug was lower than non-immune at 3.2 LD ₅₀ in 1 , 3 times and exceeded it at 11.5-13.0 LD ₅₀ 2.6-3 times, between the anti-lipid and gamma globulin values fluctuated in one direction or another 1.2-2 times.

Multiple superiority in the protective activity of anti-shigellosis immunoglobulin preparations compared with non-immune against S.typhi and Ps.aeruginosa was also experimentally shown.

Mice experiments were also performed comparing the protective activity of immunoglobulin preparations obtained by immunizing cattle with two types of vaccines. The first vaccine was obtained by the above method, the second, a comprehensive vaccine preparation, included the first vaccine and vaccine preparations obtained from Shigella strains isolated during various outbreaks of dysentery. The second component of the last vaccine preparation was a washout of the culture mass from Petri dishes. Animal experiments have shown that immunization of cattle with a second vaccine, as a rule, did not lead to an increase in the number of antibodies to the components of the Shigella cell wall and, of course, did not increase the antitoxic activity of immunoglobulin preparations. It is important to consider that living models are the most sensitive and accurate indicator of the activity of drugs in relation to various microorganisms.

In the cross-reactions of diffusion precipitation and immune electrophoresis, the antigenic relationship of cholerogen, toxogen-toxoid with exotoxins Shigella, Salmonella, Escherichia, Proteus was established. By cross-absorption of precipitates from serum to a vaccine containing cholerogen-toxoid and vaccines containing toxoids obtained from enterobacteria exotoxins, it was found that the antigenic proximity of these drugs is associated with both exotoxins and intergeneric antigens contained in them. Cross-reactions between Shigella Sonne and pseudomonads, cholera vibrios and various enterobacteria have shown that the antigenic relationship in gram-negative bacteria goes far beyond the limits of intergeneric bonds.

Clinical trials were conducted on 50 children with Sonne and Flexner 2a dysentery, salmonellosis, and coli infection caused by enterotoxigenic Escherichia coli. Assessment of the clinical efficacy of anti-shigellosis lactoglobulin included determining the timing of the disappearance of general toxic manifestations and intestinal disorders, as well as bacteriological debridement. The sanitizing effect in bacteriologically confirmed intestinal infections was obtained both in patients with a clinically severe form of the disease and in bacteriostatic agents. Stool normalization occurred in all patients, moreover, in a shorter time than with traditional methods of treatment. In no case of the use of anti-shigellosis lactoglobulin did not require the continuation of etiotropic therapy. Good tolerability of the drug, the absence of adverse reactions during its use and exacerbations of skin allergic manifestations, even in children with a weighed allergic history, were noted.

Formalin is the most popular reagent for vaccines and toxoids.

It was established that when the substances causing its detoxification are introduced into the vaccine being prepared, two processes occur: the detoxification itself and the stabilization of the structure of antigen molecules with the formation of hard bonds. In this case, chemical reactions of various complexity occur between formaldehyde and radicals and amino groups of the protein. It is still believed that for neutralization with formalin the most favorable conditions are at pH 7.2-7.3 (established traditional values).

However, to date, our Shigella vaccine, when treated with formalin in 70% of cases, has maintained a high level of toxicity or has lost immunogenicity.

The first stage of the experiment was to maintain a vaccine for immunoprophylaxis and immunotherapy of human and animal diseases caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins at 37 ° C for a year. As the final result, a high level of free (unbound) formaldehyde was maintained (0.074-0.12% of the solution amount), but the vaccine did not detoxify.

Then, the analysis of the vaccine found that at pH 7.2-7.3, the precipitate obtained in the production process of our vaccine preparation does not completely dissolve (the process is not going to the end).

If, at a pH of 7.2–7.3, only the dissolved protein is left in the solution or the pH is increased to 7.5–9.0 with a subsequent decrease to the initial level, then in any case, during storage, the protein becomes destabilized and a precipitate will precipitate. It was determined that precipitation at this stage leads to a decrease in the yield of exotoxin as a drug and, upon subsequent processing of the resulting vaccine, to a decrease in the yield of toxoid and a decrease in immunogenicity.

It was also found that the introduction of formalin at a pH of 7.2-7.3 to detoxify our vaccine preparation leads to a) additional protein destabilization and precipitation, b) a decrease in the yield of the final drug, and c) a decrease in the immunogenicity of the vaccine.

According to our research, only at pH 7.5–9.0 does: a) complete dissolution of the precipitate, b) the maximum stabilization of the protein molecules of our vaccine, c) the maximum yield of exotoxin-endotoxin as a drug, d) the maximum yield of toxoid and e) the highest rates of immunogenicity.

It was also determined that when using formalin, complete stabilization of the protein occurs only at a pH of 7.5–9.0 and a reagent of 0.4–0.8% of the amount of solution. Higher values of the reagent give a decrease in the pH of the solution and do not ensure the receipt of the claimed vaccine.

It was possible to remove the residual toxicity of the Shigella vaccine while maintaining immunogenicity only after: a) dosed addition of formalin, b) the pH of the solution was 7.5-9.0, and c) the amount of formalin 0.4-0.8% of the amount of solution. For example, initially an amount of formalin of 0.4-0.8% of the amount of solution is added, and after 2 months (keeping in an thermostat at 37 ° C), toxicity in mice is determined. If it is high, and the amount of free formaldehyde is 0.074% of the amount of the solution, then the latter is adjusted to 0.18-0.22% of the amount of solution (the amount of formalin is 0.5-0.6% of the amount of solution) and the solution is kept at 30- 60 ° C for the final conversion of exotoxin to toxoid.

As a rule, after such a procedure, the vaccine for immunoprophylaxis and immunotherapy of human and animal diseases caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins is neutralized to the necessary level while maintaining its immunogenicity. The preparation of the drug by the other aforementioned methods results in low immunogenicity of the vaccine.

It is known that the properties of any protein, in particular stability in solution at different pH values, binding to various chemicals, and its strength are determined by the presence and ratio of certain amino acids and their relative position. The maximum stability of Shigella exotoxin and toxoid, as we determined, is observed at pH 7.5-9.0.

Based on the results of studies, we can only assume that between the processes of detoxification and stabilization of the Shigella vaccine protein, there is competition for formaldehyde molecules, and soon after adding the latter to the solution. In the exotoxin molecule, strictly defined active protein groups are responsible for toxicity. The stabilization of exotoxin molecules occurs due to the fact that a significant part of the reagent displaces water from the hydration shell of the protein (according to our studies, this process is maximally expressed in a shigellosis vaccine at pH 7.5-9.0) and is associated with a weak chemical bond (otherwise formalin would not be determined). But the strength of this connection is quite sufficient to retain formaldehyde and not give it to those active protein groups that are responsible for toxicity. It does not make sense to begin the neutralization of the latter without their preliminary 100% stabilization. The detoxification process takes place in two stages. The first stage is faster (several days) and reversible, the second is extended in time and ends with the creation of stable methylene bridges. To complete the second stage and you need additional, after a certain period of time, the addition of formalin. A single increase in the concentration of formalin above the planned leads to an uneven distribution of it in the vaccine during detoxification and, ultimately, to destabilize the protein and reduce immunogenicity.

Thus, as follows from the above, it was only experimentally that we were able to find out that in order to obtain a completely neutralized highly immunogenic Shigellosis vaccine, it is first necessary to stabilize the protein to the maximum extent possible (pH of the solution is 7.5-9.0 and the amount of formalin is 0.4-0.8 % of the amount of solution), carry out preliminary detoxification of the vaccine, and then additionally introduce formalin and maintain the drug at 30-60 ° C in order to finally transfer exotoxin to toxoid.

A comprehensive study of the results of these studies led to the idea of using other substances to detoxify shigellosis exotoxin.

Recent studies conducted by us have shown that, along with formalin, benzoic acid and its salts gave similar results. Adding to the resulting vaccine benzoic acid and its salts in an amount of 0.07-4.0% of the amount of solution or their mixture in an amount of 0.1-0.3% and 0.03-2.5% of the amount of solution, and mixtures of formalin and benzoic acid or its salts in an amount of 0.1-0.3% and 0.03-2.5% of the amount of the solution, respectively, keeping the solution at a temperature of 30-60 ° C for 2 hours - 60 days led to detoxification of the drug. Immunization of animals with vaccines obtained using benzoic acid and its salts showed their high immunogenic activity.

Prior to this, benzoic acid and its salts were not used to detoxify microbial toxins and vaccines in order to obtain medical and veterinary drugs. These compounds were mainly used in the food industry as a preservative and in medicine as a local antiseptic. In nature, benzoic acid and its salts are found in cranberries.

The claimed group of inventions is united by a single inventive concept, which is aimed at obtaining various drugs for the treatment and prevention of human and animal diseases caused by gram-negative conditionally pathogenic and pathogenic microorganisms of the intestinal group and their exotoxins.

The technical result of the claimed group of inventions is to obtain a vaccine preparation containing shigellosis toxoid and S.dysenteriae corpuscular antigens, including components of microbial cells that are most closely related in antigenicity between various pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and which gives prophylactic and human vaccination therapeutic effect on various pathogenic and conditionally pathogenic gram-negative microorganisms of the intestine the second group; when immunizing farm animals with their vaccine from their milk, colostrum and blood serum, obtaining immunoglobulin preparations that have antibody activity against a wide range of pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins and, therefore, have a prophylactic and therapeutic effect against diseases, caused by the above microorganisms. An immunobiological preparation can be used to treat and prevent a wide range of diseases of the internal and external organs of humans and animals caused by various pathogenic and conditionally pathogenic gram-negative and gram-positive microorganisms and their toxins, viruses, fungi, protozoa, and helminths.

Examples

Example No. 1. A method of obtaining a vaccine for immunoprophylaxis and immunotherapy of human and animal diseases caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins. Reactor-grown culture mass of the S.dysenteriae R-form on casein broth is treated with chloroform to dissolve the cell membrane. Then centrifugation is carried out, the supernatant is taken, trichloroacetic acid is added and the pH is adjusted to 3.6. After this, centrifugation is carried out, the resulting precipitate is dissolved in buffer, the pH of the resulting solution is adjusted to 8.3. In order to obtain anatoxin, formalin is added to the resulting preparation in an amount of 0.5% of the solution at a vaccine pH of 8.3 and the resulting solution is kept at a temperature of 37 ° C in a thermostat for 1-2 months until the exotoxin is completely neutralized. If the toxicity persists, then the vaccine is sent for further processing by adjusting the concentration of free formaldehyde to 0.18% of the amount of the solution at the pH of the vaccine solution 8.3 and keeping it in the thermostat for an additional 1-2 months. Moreover, the resulting vaccine contains protein - 10-15 mg / ml, nucleic acids - 0.2-0.5 mg / ml, lipids - traces. Then produce packaging and corking the resulting vaccine.

After that, the vaccine preparation is used to immunize humans and animals for the purpose of immunoprophylaxis and immunotherapy of intestinal infectious diseases caused by gram-negative pathogenic and opportunistic microorganisms of the intestinal group and their exotoxins, as well as vaccination of farm animals in order to obtain immune raw materials for the production of immunoglobulin preparations.

Example No. 2. A method of obtaining a vaccine for immunoprophylaxis and immunotherapy of human and animal diseases caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins. Reactor-grown culture mass of the S.dysenteriae R-form on casein broth is treated with chloroform to dissolve the cell membrane. Then centrifugation is carried out, the supernatant is taken and the pH is adjusted to 3.2 with trichloroacetic acid. After that, centrifugation is carried out in order to obtain a precipitate containing corpuscular antigens and shigellosis exotoxin, dissolve the precipitate in a buffer, and adjust the pH of the resulting solution to 8.6. In order to obtain anatoxin, benzoic acid is added to the resulting preparation in an amount of 1.0% of the solution at a vaccine solution pH of 8.6 and the resulting solution is kept in an thermostat at a temperature of 37 ° C for 1-2 months until the exotoxin is completely neutralized. Moreover, the resulting vaccine contains protein - 10-15 mg / ml, nucleic acids - 0.2-0.5 mg / ml, lipids - traces. Then produce packaging and capping or packaging and lyophilization of the resulting vaccine.

After that, the vaccine preparation is used to immunize humans and animals for the purpose of immunoprophylaxis and immunotherapy of intestinal infectious diseases caused by gram-negative pathogenic and conditionally pathogenic microorganisms of the intestinal group and their exotoxins, as well as vaccination of farm animals to obtain immunoglobulin preparations.

Example No. 3. A method of obtaining a vaccine for immunoprophylaxis and immunotherapy of human and animal diseases caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins. The vaccine is obtained according to example No. 1. Before packaging, the vaccine is freeze dried to obtain powders.

Example No. 4. A method of obtaining an immunoglobulin preparation. After immunization of farm animals with a vaccine for immunoprophylaxis and immunotherapy of human and animal diseases caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins obtained according to Example No. 1, blood sampling is performed to obtain an immunoglobulin preparation from blood serum.

In order to obtain the drug, 1 M acetic acid is added to the blood serum of cattle to a solution pH of up to 4.3. At a temperature of 22 ° C with vigorous stirring, caprylic acid is added at the rate of 3.0 ml per 100 ml of solution. After stopping the reagent addition, stirring is continued for 1-2 hours. Then, centrifugation is carried out, after which the precipitate is removed, and the immunoglobulins remain in solution, the pH of the solution is adjusted to 7.01 M sodium hydroxide with vigorous stirring, and the resulting preparation is dialyzed against physiological sodium chloride solution. The immunoglobulin preparation obtained in this way contains the fraction of immunoglobulins G, A and M in a ratio of (45: 1: 4) and has antibody activity against a wide range of pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins and, therefore, has preventive and therapeutic effect in relation to the listed microorganisms.

Example No. 5. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum (IgG: IgA: IgM) (50: 1: 5) - 0.25 g Vitamin A - 1650 ME a-Tocopherol acetate (vitamin E) - 0.005 g Ascorbic Acid (Vitamin C) - 0.05 g Fructose - 0.2 g Technological components for tabletting (talc and calcium stearate) - to tablet mass   0.55 g

The drug gave positive results when tested on 10 volunteers with dysentery.

The immunoglobulin fraction isolated from the blood serum of cows immunized with a vaccine for immunoprophylaxis and immunotherapy of human and animal diseases caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins gave pronounced antitoxic and antibacterial effects. Due to the presence of antitoxic antibodies in the preparation, treatment was carried out with less pronounced intoxication than with traditional methods of treatment. After the end of the course of treatment, there was no violation of the normal intestinal microflora observed during treatment with antibiotics and chemotherapy.

The introduction of vitamins A and E into the tablet for the purpose of antioxidant therapy reduces the level of peroxidic compounds, which increases during the disease, increases the reparative processes in the intestinal mucosa, which has a positive effect on well-being and led to a faster recovery. In addition, vitamin A restores the activity of vitamin E in the body. Vitamin C is added to increase tissue regeneration and reduce intoxication.

Example No. 6. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 0.1 g Immunoglobulin fraction human blood serum - 0.1 g Bifidobacteria - 0.007 g Vitamin A - 1650 ME α-Tocopherol Acetate (Vitamin E) - 0.005 g Ascorbic Acid (Vitamin C) - 0.05 g Pyridoxine Hydrochloride (Vitamin B6) - 0.005 g Potassium dihydrogen phosphate (KH ₂ PO ₄ ) - 0.0052 g Skimmed milk powder - 0.2 g Technological components for tabletting (talc and calcium stearate) - to the mass of tablets   0.55 g

Example No. 7. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.2 g α-Tocopherol Acetate (Vitamin E) - 0.01 g Vitamin A - 1650 ME Bifidobacteria - 0.1 g Lactobacilli - 0.1 g Fructose - 0.05 g Technological components for tabletting - to the mass of tablets   0.55 g

The drug gave positive results when tested on 12 volunteers with dysentery and coli infection.

The immunoglobulin fraction of colostrum of cows vaccinated with a vaccine for the immunoprophylaxis and immunotherapy of human and animal diseases caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins gave a pronounced antitoxic and antibacterial effect. Due to the presence of antitoxic antibodies in the preparation, treatment was carried out with less pronounced intoxication than with traditional methods of treatment. Moreover, an increase in the dose of the drug led to a more rapid disappearance of general toxic manifestations, intestinal disorders and bacteriological debridement. After the end of the course of treatment, there was no violation of the normal intestinal microflora observed during treatment with antibiotics and chemotherapy.

The introduction of bifidobacteria and lactobacilli into the tablet led to a more rapid normalization of intestinal normoflora than with traditional methods of treatment. Antioxidant therapy with vitamins A and E, as you know, reduces the level of peroxidic compounds that increase during the disease, increases the reparative processes in the intestinal mucosa, which positively affected the well-being of patients and led to a faster recovery.

Example No. 8. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction, derived from colostrum of a cow, immunized vaccine for immunoprophylaxis and immunotherapy diseases of humans and animals, caused by pathogenic and conditionally pathogenic gram-negative gut microorganisms and their exotoxins - 0.2 g Cow immunoglobulin G with antirotoviral activity - 0.2 g α-Tocopherol Acetate (Vitamin E) - 0.01 g Vitamin A - 1650 ME Lactoferrin - 0.1 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 9. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 0.4 g Lactoperoxidase - 0.05 g Lactoferrin - 0.05 g Bifidobacteria - 0.02 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 10. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 0.4 g Brewer's yeast - 0.1 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 11. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction goat colostrum - 0.4 g Lactoperoxidase - 0.05 g Lysozyme - 0.05 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 12. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum sheep - 0.4 g Baker's yeast - 0.05 g Activated carbon - 0.07 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 13. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction ram blood serum - 0.4 g Activated carbon - 0.1 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 14. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 0.4 g Orozomucoid - 0.05 g Fructose - 0.05 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 15. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction goat colostrum - 0.4 g Haptoglobin - 0.1 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 16. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.4 g Tetracycline hydrochloride - 0.1 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 17. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 0.25 g Sulfadimezin - 0.25 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 18. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 0.4 g Chloramphenicol - 0.05 g Nystatin - 0.05 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 19. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 0.4 g Cycloserine - 0.1 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 20. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum sheep - 0.48 g Furazolidone - 0.02 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 21. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction ram blood serum - 0.4 g Ribamidide - 0.1 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 22. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.4 g Calcium lactate - 0.1 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 23. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 0.2 g Human immunoglobulin G - 0.2 g Methylmethionine sulfonium chloride - 0.02 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 24. A vaccine preparation in tablet form. To obtain a vaccine preparation, the following components are used:

Immunization vaccine and immunotherapy of diseases humans and animals caused by pathogenic and conditionally pathogenic gram-negative microorganisms intestinal group and their exotoxins - 0.45 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 25. A multicomponent vaccine preparation in tablet form. To obtain a multicomponent vaccine preparation, the following components are used:

Immunization vaccine and immunotherapy of diseases human and animal caused pathogenic and conditionally pathogenic gram negative gut microorganisms groups and their exotoxins - 0.2 g Rotovirus vaccine - 0.25 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 26. A multicomponent vaccine preparation in tablet form. To obtain a multicomponent vaccine preparation, the following components are used:

Immunization vaccine and immunotherapy of diseases human and animal caused pathogenic and conditionally pathogenic gram-negative microorganisms intestinal group and their exotoxins - 0.45 g Typhoid Vi antigen - 0.05 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 27. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.5 g Methylmethionine sulfonium chloride - 0.01 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 28. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 0.4 g Peppermint - 0.1 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 29. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.2 g Serum Immunoglobulin G blood sheep - 0.2 g Tea mushroom - 0.1 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 30. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 0.4 g Pantocrine - 0.1 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 31. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 0.25 g Human immunoglobulin A - 0.2 g Arachidonic acid - 0.025 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 32. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.5 g Lipoic acid - 0.005 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 33. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.5 g Arginine - 0.005 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 34 An immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 0.5 g Caffeine - 0.01 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 35. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 0.45 g Panaxoside - 0.05 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 36. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.45 g Griseofulvin - 0.05 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 37. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 0.45 g Clotrimazole - 0.05 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 38. Immunobiological preparation in tablet form. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.3 g Methyluracil - 0.2 g Technological components for tabletting - to tablet mass   0.55 g

Example No. 39. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.1 g Lactobacilli - 10 ⁶ CFU α-Tocopherol Acetate (Vitamin E) - 0.01 g

The basis for the suppository (emulsifier T-2,

 fat for chocolate, paraffin) - to mass suppository 1.4 g

The drug gave positive results in the treatment of vaginal infection caused by gram-negative pathogenic enterobacteria.

The immunoglobulin fraction of colostrum of cows vaccinated with a vaccine for immunoprophylaxis and immunotherapy of human and animal diseases caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins gave a pronounced sanitizing effect. Moreover, unlike traditional methods of treatment with antibiotics and chemotherapy, the products of the death of microorganisms enter the bloodstream minimally, and the irritation of the mucosa disappears faster.

Lactobacilli and vitamin E contributed to faster reparative processes of the mucosa and the restoration of the internal environment of the vagina.

Example No. 40. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 0.05 g Immunoglobulin fraction human blood serum - 0.05 g α-Tocopherol Acetate (Vitamin E) - 0.01 g Chloramphenicol - 0.02 g The basis for the suppository - to mass (cocoa butter, gelatin mass) suppository 1.4 g

Example No. 41. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.1 g Human immunoglobulin G - 0.05 g Chamomile flower extract - 0.02 g The basis for the suppository - to mass (cocoa butter, lanolin) suppository 1.4 g

The drug gave positive results in the treatment of vaginal infection caused by gram-positive bacteria, gram-negative enterobacteria and mixed infection.

The immunoglobulin fraction of colostrum of cows immunized with a vaccine for the immunoprophylaxis and immunotherapy of human and animal diseases caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins, and human IgG isolated from blood serum or waste from gamma-globulin production, gave a good sanitizing effect.

Chamomile flower extract has anti-inflammatory and antiseptic properties, weakens allergic reactions, enhances the regeneration of the mucosa.

Example No. 42. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 0.15 g Furazolidone - 0.02 g The basis for the suppository - to mass (butyrol) suppository 1.4 g

Example No. 43. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 0.18 g Fructose - 0.01 g Belladonna extract - 0.01 g

The basis for the suppository

(gelatin, glycerin, water) - to mass suppository 1.4 g

Example No. 44. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction milk obtained from a cow, immunized vaccine for immunoprophylaxis and immunotherapy diseases of humans and animals, caused by pathogenic and conditionally pathogenic gram-negative gut microorganisms and their exotoxins - 0.1 g Human immunoglobulin G with antirotoviral activity - 0.1 g Suppository base - to mass   suppository 1.4 g

Example No. 45. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 0.18 g Lactoperoxidase - 0.02 g Suppository base - to the mass of the suppository   1.4 g

Example No. 46. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.18 g Brewer's yeast - 0.02 g Suppository base - to mass   suppository 1.4 g

Example No. 47. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.085 g Immunoglobulin fraction goat colostrum - 0 085 g Methylmethionine sulfonium chloride - 0.01 g Orozomucoid - 0.02 g Suppository base - to mass   suppository 1.4 g

Example No. 48. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.17 g Human interferon - 0.03 g Suppository base - to mass   suppository 1.4 g

Example No. 49. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 0.15 g Kombuka tea - 0.05 g Suppository base - to mass   suppository 1.4 g

Example No. 50. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction bull serum - 0.15 g Skim milk of a person - 0.05 g Suppository base - to mass   suppository 1.4 g

Example No. 51. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 0.05 g Immunoglobulin fraction pig blood serum - 0.05 g Royal jelly - 0.1 g Suppository base - to mass   suppository 1.4 g

Example No. 52. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.05 g Immunoglobulin G goat milk - 0.05 g Activated carbon - 0.1 g Suppository base - to mass   suppository 1.4 g

Example No. 53. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction

goat colostrum - 0.1 g Chloramphenicol - 0.1 g Suppository base - to mass   suppository 1.4 g

Example No. 54. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 0.15 g Norsulfazole - 0.05 g Suppository base - to mass   suppository 1.4 g

Example No. 55. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.15 g Chlorchinaldol - 0.05 g Suppository base - to mass   suppository 1.4 g

Example No. 56. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction goat milk - 0.1 g Chlorophyllipt - 0.1 g Suppository base - to mass   suppository 1.4 g

Example No. 57. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.1 g Ethionamide - 0.1 g Suppository base - to mass   suppository 1.4 g

Example No. 58. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.15 g Ganciclovir - 0.03 g Suppository base - to mass   suppository 1.4 g

Example No. 59. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 0.19 g Ubinon - 0.01 g Suppository base - to mass   suppository 1.4 g

Example No. 60. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 0.1 g Ferrous iron lactate - 0.1 g Suppository base - to mass   suppository 1.4 g

Example No. 61. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 0.15 g Fructose - 0.05 g Suppository base - to mass   suppository 1.4 g

Example No. 62. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction buffalo milk - 0.15 g Peanut butter - 0.05 g Suppository base - to mass   suppository 1.4 g

Example No. 63. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 0.09 g Deer immunoglobulin G - 0.09 g Arginine - 0.02 g Suppository base - to mass   suppository 1.4 g

Example No. 64. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction sheep blood serum - 0.15 g Lipoic acid (sodium salt) - 0.05 g The basis for the suppository - to mass suppository 1.4 g

Example No. 65. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction

bull serum - 0.15 g Caffeine - 0.05 g The basis for the suppository - to mass   suppository 1.4 g

Example No. 66. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction goat blood serum - 0.15 g Panaxoside - 0.05 g The basis for the suppository - to mass   suppository 1.4 g

Example No. 67. Vaccine preparation in the form of a suppository. To obtain a vaccine preparation, the following components are used:

Immunization vaccine and immunotherapy of human diseases and animals caused pathogenic and conditionally pathogenic gram-negative microorganisms intestinal group and their exotoxins - 0.2 g Suppository base - to mass   suppository 1.4 g

Example No. 68. Multicomponent vaccine preparation in the form of a suppository. To obtain a multicomponent vaccine preparation, the following components are used:

Immunization vaccine and immunotherapy of human diseases and animals caused by pathogenic and opportunistic gram-negative microorganisms intestinal group and their exotoxins - 0.1 g Rotovirus vaccine - 0.1 g Suppository base - to mass   suppository 1.4 g

Example No. 69. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.15 g Griseofulvin - 0.05 g Suppository base - to mass   suppository 1.4 g

Example No. 70. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 0.15 g Clotrimazole - 0.05 g Suppository base - to mass   suppository 1.4 g

Example No. 71. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction bull serum - 0.1 g Reogluman - 0.1 g Suppository base - to mass   suppository 1.4 g

Example No. 72. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 0.1 g Enterodesus - 0.1 g Suppository base - to mass   suppository 1.4 g

Example No. 73. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 0.1 g Wolek - 0.1 g Suppository base - to mass   suppository 1.4 g

Example No. 74. Immunobiological preparation in the form of a suppository. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 0.1 g Methyluracil - 0.1 g Suppository base - to mass   suppository 1.4 g

Example No. 75. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 5.0 g Immunoglobulin fraction sheep blood serum - 5.0 g Oil and fat base (emulsifier T-2, creamy - 90.0 g oil, solvent)

Example No. 76. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 5.0 g Immunoglobulin fraction human blood serum - 5.0 g Oil and fat base - 90.0 g (petroleum jelly, petroleum jelly)

Example No. 77. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 5.0 g Human immunoglobulin G - 5.0 g Oil and fat base (lanolin, petroleum jelly, water) - 90.0 g

Example No. 78. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 15.0 g Oil and fat base - 85.0 g (petroleum jelly, starch)

Example No. 79. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 5.0 g Lactoferrin - 5.0 g Oil and fat base - 90.0 g (petroleum jelly, water)

The drug gave positive results in the healing of wounds of various origins.

The immunoglobulin fraction obtained from colostrum of cows immunized with a vaccine for the immunoprophylaxis and immunotherapy of human and animal diseases caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins contains antibodies to exotoxins and corpuscular antigens of gram-negative microorganisms, as well as significant spectrum antibodies to gram-positive microorganisms.

Lactoferrin has a pronounced bacteriostatic effect.

Example number 80. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 5.0 g Lysozyme - 5.0 g Oil and fat base - 90.0 g (butter, petroleum jelly, paraffin, water)

Example No. 81. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 9.0 g Bifidobacteria - 10 ⁷ CFU Oil and fat base - 90.0 g

Example No. 82. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction goat milk - 5.0 g Brewer's yeast - 5.0 g Oil and fat base - 90.0 g

Example No.83. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction ram blood serum - 5.0 g Vitamin A - 1.0 g α-tocopherol acetate - 2.0 g Riboflavin (Vitamin B ₂ ) - 2.0 g Oil and fat base - 90.0 g

Example No. 84. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 5.0 g Methylmethionine sulfonium chloride - 5.0 g Oil and fat base - 90.0 g

Example No. 85. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 5.0 g Orozomucoid - 5.0 g Oil and fat base (butter, petroleum jelly, paraffin, water)                             - 90.0 g

The drug gave positive results in the treatment of various wounds, wound infections and burns.

The immunoglobulin fraction obtained from the blood serum of cows immunized with a vaccine for immunoprophylaxis and immunotherapy of diseases of humans and animals caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins contains antibodies to exotoxins of gram-negative microorganisms, as well as a significant spectrum of natural microorganisms to grams .

Orozomucoid has a pronounced wound healing effect.

Example No. 86. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 5.0 g Human intferon leukocyte - 5 million ME Oil and fat base - 90.0 g

Example No. 87. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 5.0 g Aloe extract - 5.0 g Oil and fat base - 90.0 g

Example No. 88. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 5.0 g Sugar Laminaria Extract - 5.0 g Oil and fat base - 90.0 g

Example No. 89. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 5.0 g Salmon Fish Milk Extract - 5.0 g Oil and fat base - 90.0 g

Example No. 90. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 5.0 g Propolyus - 5.0 g Oil and fat base - 90.0 g

Example No. 91. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction bull serum - 5.0 g Chloramphenicol - 5.0 g Oil and fat base - 90.0 g

Example No. 92. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 5.0 g Streptocide - 5.0 g Oil and fat base - 90.0 g

Example No. 93. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 5.0 g Nitroxoline - 5.0 g Oil and fat base - 90.0 g

Example No. 94. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 5.0 g Sodium Usninat - 5.0 g Oil and fat base - 90.0 g

Example No. 95. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 3.0 g Streptomycin sulfate - 5.0 g Oil and fat base - 90.0 g

Example No. 96. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 5.0 g Acyclovir - 5.0 g Oil and fat base - 90.0 g

Example No. 97. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 5.0 g Griseofulvin - 5.0 g Oil and fat base - 90.0 g

Example No. 98. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 5.0 g Clotrimazole - 5.0 g Oil and fat base - 90.0 g

Example No. 99. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 3.0 g Ubinon - 5.0 g Oil and fat base - 90.0 g

Example No. 100. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 5.0 g Calcium lactate - 5.0 g Oil and fat base - 90.0 g

Example No. 101. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 5.0 g Pectin - 5.0 g Oil and fat base - 90.0 g

Example No. 102. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 5.0 g Sea buckthorn oil - 10.0 g Oil and fat base - 85.0 g

Example No. 103. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 5.0 g Arginine - 5.0 g Oil and fat base - 90.0 g

Example No. 104. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction bull serum - 5.0 g Lipoic acid - 5.0 g Oil and fat base - 90.0 g

Example No. 105. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 5.0 g Viburnin - 5.0 g Oil and fat base - 90.0 g

Example No. 106. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 5.0 g Eleutheroside - 5.0 g Oil and fat base - 90.0 g

Example No. 107. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction colostrum cow - 5.0 g Reogluman - 5.0 g Oil and fat base - 90.0 g

Example No. 108. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 5.0 g Enterodesus - 5.0 g Oil and fat base - 90.0 g

Example No. 109. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow's milk - 5.0 g Wolek - 5.0 g Oil and fat base - 90.0 g

Example No. 110. Vaccine preparation in the form of ointments. To obtain a vaccine preparation, the following components are used:

Immunization vaccine and immunotherapy of diseases human and animal caused pathogenic and conditionally pathogenic gram-negative microorganisms intestinal group and their exotoxins     - 10.0 g Oil and fat base - 90.0 g

Example No. 111. A multicomponent vaccine preparation in the form of ointments. To obtain a multicomponent vaccine preparation, the following components are used:

Immunization vaccine and immunotherapy of diseases humans and animals caused by pathogenic and opportunistic gram-negative microorganisms intestinal group and their exotoxins     - 5.0 g Staphylococcal vaccine - 3.0 g Oil and fat base - 90.0 g

Example No. 112. Immunobiological preparation in the form of ointments. To obtain an immunobiological preparation, the following components are used:

Immunoglobulin fraction cow serum - 5.0 g Methyluracil - 5.0 g Oil and fat base - 90.0 g

A vaccine preparation obtained from S.dysenteriae R-form strains and including shigellous toxoid and corpuscular antigens has high immunogenicity. Due to the presence of antigens common to gram-negative microorganisms in the vaccine, a wide range of antibodies to pathogenic microorganisms and their exotoxins are produced during immunization of animals, which is confirmed in vitro and in vivo. The ineffectiveness of commercial immune immunoglobulin preparations - anti-lipoprotein lactoglobulins and against opportunistic bacteria and salmonella against shigella and shigellosis exotoxin, as well as the possibility of using these lactoglobulin drugs instead of placebo in the in vivo study of antibacterial and antitoxic activity of anti-shigella immunoglobulins was shown.

It is proposed to use this vaccine preparation for the prevention of diseases caused by pathogenic and conditionally pathogenic gram-negative microorganisms, especially the intestinal group, as well as for the production of immunoglobulin preparations of a wide range of antibacterial and antitoxic activity from colostrum, milk and blood of immune animals.

An original method is described for the neutralization of a semi-finished shigellosis vaccine consisting of exotoxin and corpuscular antigens in order to obtain a vaccine preparation using benzoic acid and its salts, as well as when used together with formalin.

Variants of immunobiological preparations are proposed for the combination of this vaccine with other vaccines, as well as immunoglobulins and immunoglobulin preparations with various biologically active substances and preparations of animal, plant and synthetic origin.

The claimed group of inventions is united by a single inventive concept, which is aimed at immunoprophylaxis and immunotherapy of human and animal diseases caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins.

Claims (13)

1. A method of obtaining a vaccine for immunoprophylaxis and immunotherapy of human and animal diseases caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins, characterized in that it involves the cultivation of S.dysenteriae R-forms, cell lysis by treatment with chloroform, centrifugation with obtaining a supernatant, then the supernatant is treated with saturated monobasic carboxylic acids or their derivatives to pH 3.0-5.0, centrifugation to obtain a precipitate, soda containing corpuscular antigens and shigellosis exotoxin, dissolving the precipitate in buffer and adjusting the pH to 7.5–9.0, adding formalin in an amount of 0.4–0.8% of the amount of solution, or benzoic acid, or its salts in an amount of 0, 07-4.0% of the amount of solution, or a mixture thereof, while the amount of formalin is 0.1-0.3% of the amount of solution, the amount of benzoic acid or its salts 0.03-2.5% of the amount of solution, keeping the solution at a temperature of 30-60 ° C for 2 hours - 60 days before the transition of exotoxin to toxoid to obtain a vaccine, then t packaging and corking. 2. The method according to claim 1, characterized in that before packaging the final product is subjected to freeze drying to obtain granules or powders. 3. The method according to claim 1, characterized in that before packaging the ointment base or suppository base is administered into the vaccine. 4. A method of obtaining a vaccine for immunoprophylaxis and immunotherapy of human and animal diseases caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins, characterized in that it involves the cultivation of S.dysenteriae R-forms, cell lysis by treatment with chloroform, centrifugation with obtaining a supernatant, then the supernatant is treated with saturated monobasic carboxylic acids or their derivatives to pH 3.0-5.0, centrifugation to obtain a precipitate, soda containing corpuscular antigens and shigellosis exotoxin, dissolving the precipitate in buffer and adjusting the pH to 7.5-9.0, adding formalin in an amount of 0.4-0.8% of the amount of solution or benzoic acid, or its salts in an amount of 0.07 -4.0% of the amount of solution, or a mixture thereof, while the amount of formalin is 0.1-0.3% of the amount of solution, and the amount of benzoic acid or its salts 0.03-2.5% of the amount of solution, aging solution at a temperature of 30-60 ° C for 2 hours - 60 days, after which additional formalin treatment is carried out until I free formaldehyde in a solution of 0.18-0.25% of the amount of the solution, or benzoic acid, or its salts or their mixture, to the content of free benzoic acid or salts of benzoic acid in the solution of 0.07-4.0% of the amount of solution the amount of free formaldehyde in the solution is 0.037-0.12% of the amount of the solution, and the amount of free benzoic acid or its salts in the solution is 0.03-2.5% of the amount of the solution, keeping at 30-60 ° C for 2 hours - 30 days before the transition of exotoxin to toxoid to obtain a vaccine, then p sfasovku and capping. 5. The method according to claim 4, characterized in that before packaging the ointment base or suppository base is administered into the vaccine. 6. The method according to claim 4, characterized in that before packaging the final product is subjected to freeze drying to obtain granules or powders. 7. A vaccine for immunoprophylaxis and immunotherapy of diseases of humans and animals caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins, characterized in that obtained by the method according to any one of claims 1 to 6. 8. A method of producing an immunoglobulin preparation for the prevention and treatment of diseases of humans and animals caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins, characterized in that it provides immunization of animals with a vaccine for immunoprophylaxis and immunotherapy of human and animal diseases caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins obtained by the method according to any one of claims 1 to 6, a fence at animals of immune milk, and / or colostrum, and / or blood, obtaining an immunoglobulin fraction, sterilization, packaging and capping. 9. A method of producing an immunoglobulin preparation according to claim 8, characterized in that after sterilization, drying is carried out. 10. An immunoglobulin preparation for the prevention and treatment of diseases of humans and animals caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins, characterized in that it is obtained by the method according to any one of claims 8 to 9. 11. The immunoglobulin preparation according to claim 10, characterized in that it further comprises at least one component selected from the series: flavoring agents, aromatic additives, a base for suppositories, a base for ointment forms, processing aids for tableting. 12. An immunobiological preparation for the prevention and treatment of diseases of humans and animals caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and exotoxins, characterized in that it contains an immunoglobulin preparation obtained by the method of claim 8, and at least two components, selected from the series: lactoferrin, enzymes, proteolytic enzyme inhibitors, human and / or animal normoflora preparations, yeast, vitamins, vitamin-like substances, human acute phase proteins and / or here, human and / or animal cytokines, components of higher plants, components of lower plants, components of natural products, beekeeping products, enterosorbents, antibiotics, antimicrobial chemotherapy drugs, sulfa drugs, antimicrobial and antiparasitic drugs of natural origin, anti-tuberculosis drugs, antiviral drugs, antifungal antibiotics , synthetic antifungal drugs, metabolic stimulants, antioxidants, mineral d formulations, carbohydrates, lipids, essential and / or essential amino acids, organic acids, alkaloids, glycosides, flavorings, aromatic additives, base for suppositories, base for ointment forms, technological additives for tabletting. 13. A multicomponent vaccine for the prevention and treatment of diseases of humans and animals caused by pathogenic and conditionally pathogenic gram-negative microorganisms of the intestinal group and their exotoxins, characterized in that it contains, at least as one of the components, a vaccine obtained by the method according to any one of claims .1-6.