Food, cosmetic and pharmaceutical formulation with an immunomodulatory and protective anti-viral effect

Abstract

Field of application: the invention relates to biotechnology, can be used in the food industry, medicine and cosmetology in the prevention and/or treatment of diseases associated with immunodeficiencies and respiratory viral infections. The proposed food, cosmetic and pharmaceutical composition with an immunomodulating and protective antiviral effect additionally has the ability to completely prevent infection by a number of viruses, including influenza, rotavirus and coronavirus, as well as stimulate the immune system more than individual probiotics.

Claims

1. A food, cosmetic or pharmaceutical composition comprising: a. a mixture of probiotics with an immunomodulating effect, comprising modified spores of the probiotic bacterium B. coagulans, the surface of which is covalently chemically modified by acylation with dicarboxylic and polycarboxylic acid anhydrides or halides to a degree of up to 15% by weight of the spores; b. papaverine or a raw material containing papaverine; c. plant extracts comprising glycosides, amino acids, and vitamins; and d. at least two components selected from gibberellin, para-aminobenzoic acid, carnitine, and an auxin.

2. The composition according to claim 1, further comprising probiotic fungus S. boulardii in an amount of from 1 to 10 lg CFU/ml per dose of the composition.

3. The composition according to claim 1, wherein the plant extracts with glycosides includes a licorice extract in an amount of from 0.05% to 0.5% by weight of the composition.

4. The composition according to claim 1, wherein the plant extracts with glycosides includes a Stevia extract in an amount of from 0.05% to 0.5% by weight of the composition.

5. The composition according to claim 1, wherein the plant extracts with glycosides includes a spirulina extract in an amount of from 0.05% to 5% by weight of the composition.

6. The composition according to claim 1, wherein the plant extracts with glycosides includes an alfalfa extract in an amount of from 0.05% to 5% by weight of the composition.

7. The composition according to claim 1, wherein the vitamins include cyanocobalamin.

8. The composition according to claim 1, wherein the vitamins include riboflavin.

9. The composition according to claim 1, wherein the vitamins include thiamine.

10. The composition according to claim 1, wherein the vitamins include folic acid.

11. The composition according to claim 1, wherein the plant extracts further comprise rutin.

12. The composition according to claim 1, wherein the vitamins include a mixture of cyanocobalamin, riboflavin, thiamine, folic acid, and rutin.

13. The composition according to claim 1, wherein the amino acids include arginine.

14. The composition according to claim 1, wherein the amino acids include taurine.

15. The composition according to claim 1, wherein the amino acids include lysine.

16. The composition according to claim 1, wherein the amino acids include a mixture of arginine, taurine and lysine.

17. The composition according to claim 1, wherein the modified spores of the probiotic bacterium B. coagulans are present in an amount of from 1 to 10 lg CFU/ml per dose of the composition.

18. The composition according to claim 1, wherein the raw material containing papaverine includes crushed ripe poppy seeds.

19. The composition according to claim 1, wherein the raw material containing papaverine includes an extract of mature poppy seeds.

20. The composition according to claim 1, wherein the compounds of the benzylisoquinoline group include papaverine.

21. The composition according to claim 1, comprising gibberellin in an amount of 0.01-0.05% by weight of the composition.

22. The composition according to claim 1, comprising para-aminobenzoic acid in an amount of 0.01-0.5% by weight of the product.

23. The composition according to claim 1, comprising carnitine in an amount of 0.1-1.0% by weight of the composition.

24. The composition according to claim 1, comprising the auxin in an amount of 0.01-0.05% by weight of the composition.

Description

EXAMPLES

(1) The following examples describe some embodiments of the current invention. However, it should be taken in to consideration that these examples are for illustration purposes only and do not in any way limit the scope of the invention.

Example 1. Obtaining Covalent Chemically Modified Spores of B. coagulans (KCMS)

(2) Any Probiotic Strain of B. coagulans can be Used to Obtain Spores.

(3) 1. Inoculate the culture, for example, strains of Bacillus coagulans MTCC 5856, Bacillus coagulans ATCC 31284, Bacillus coagulans NBRC 3887, Bacillus coagulans ATCC 7050 or any other in 1.0 liter of broth yeast extract with glucose and acetate (HiMedia, Mumbai, India) or MRS broth nutrient medium containing 0.5% Tween-80, or in an environment based on corn extract powder and incubated at 37° C. for 24-48 hours;

(4) 2. After incubation and accumulation of bacterial biomass in the first stage the temperature reduced of the cultivated medium to 5° C. and carry out incubation at this temperature for 24-48 hours. At the same time, there is complete sporulation of all bacteria in the nutrient medium due to the temperature-negative bacteria conditions.

(5) 3. The nutrient medium is centrifuged at 20 g for 2 hours, the spore precipitate is washed twice with distilled water. A spore suspension is obtained in a ratio of dry sediment to water of 1:100. As a result of the suspension containing purified spores, residual nitrogen is determined and converted to protein by any classical method known to a person skilled in the area.

(6) 4. Dry succinic anhydride is added to the spore suspension in an amount up to 300% by weight of dry spores (for example, 300 mg of succinic anhydride is added per 100 mg of dried spores) and the solution is stirred until the precipitate of succinic anhydride is completely dissolved. Modification of more than 300% of the dry mass of spores leads to a loss in spores of viability and ability to germinate, they completely die. Instead of succinic anhydride, other carboxylic and polycarboxylic acid anhydrides and halides may be used. Instead of the acylation process with succinic anhydride, an alkylation process using haloalkanes or halogen derivatives of carboxylic and polycarboxylic acids can be used. Modification should lead to an increase in the total negative charge of the spore surface. This enhances the adhesion of spores to the intestinal wall, enhances the phagocytosis of such spores by macrophages and causes a more active immune response in shorter terms.

(7) 5. Next, the modified spore solution is lyophilized or dried on a tray at a temperature of up to 40° C. and then used to obtain the composition according to the claimed invention.

Example 2. Obtaining a Composition Based on Covalent Chemically Modified Spores of B. Coagulans (KCMSB) for Inclusion in Food, Cosmetic and Pharmaceutical Products

(8) By using any mixer for solid compositions, we will conduct the manufacturing of the composition according to the invention. Dry KCMSB with an activity of 1-50 lg CFU/g is added to the mixer, dry lyophilized powder S. boulardii is added in an amount of 1 to 50 lg CFU/g, licorice extract is added in an amount of 0.05% to 0.5% of the total the weight of the composition, add Stevia extract in an amount of from 0.05% to 0.5% of the total weight of the composition, add spirulina extract in an amount of from 0.05% to 5% of the total weight of the composition, add alfalfa extract in an amount of from 0.05% to 5% by weight of the product, add crushed mature poppy seeds or poppy extract or papaverine in an amount of from 0.05% to 5% by weight of the composition, add gibberellin in an amount of 0.01-0.05% by weight of the composition, add carnitine in an amount of 0.1-1, 0% by weight of the composition. Also, in addition to gibberellin and carnitine, the composition may include para-aminobenzoic acid in an amount of 0.01-0.5% by weight of the composition and auxin or its derivative in an amount of 0.01-0.05% by weight of the composition. The mixed composition is mixed in a mixer until it smooth and used in the future to obtain food and pharmaceutical compositions, toothpaste and cosmetic creams.

(9) Determination of Germination Rate of B. coagulans (BC) Spores

(10) KCMSB in the form of a suspension in the amount of 0.1 g/1 ml of distilled water is applied to the surface of six Petri dishes with Lacto medium. A similar procedure is carried out with different KCMSB compositions (presented in Table 1), 6 Petri dishes per sample. Unmodified spores are applied to the last group of Petri dishes. Every 3 hours, a smear sample is taken, stained according to Gram and the presence and quantity of colony forming units—vegetative sprouted forms of Bacillus coagulans (BC)—are determined. The results are presented in table 2.

(11) TABLE-US-00002 TABLE 2 Results of determining the rate of KCMSB germination in comparison with unmodified Bacillus coagulans spores The time of 1 g CFU/ml Group appearance of the 72 hours No. The name and composition spores first colonies later 1. Lactovit-Forte (unmodified spores) 72 ± 5  15 ± 5  2. KCMSB (option with poppy seed 36 ± 2* 85 ± 12* extract) 3. KCMSB (papaverine variant) 36 ± 6* 90 ± 16* 4. KCMSB (poppy extract variant, 24 ± 4* 90 ± 15* gibberellin, carnitine)) 5. KCMSB (modified disputes only) 48 ± 3* 65 ± 10* *P < 0.01, the difference is significant against the control of group 1

(12) As can be seen from table 2, the germination of vegetative forms of BC from all HCMSB compositions is statistically significantly faster than the control. At 72 hours of incubation in the control, only 15 lg CFU/ml of CD is observed, whereas in the experimental groups this indicator is 6 to 9 times higher. This indicates that the components of HCMSB compositions stimulate germination of CD spores and stimulate further growth of germinated bacteria. The absence of benzylisoquinolines (papaverine) in the composition leads to less effective stimulation of CD growth, although it is higher than the control (65±10 lg CFU/ml)

Example 3. Obtaining Food, Cosmetic and Pharmaceutical Products Based on the Composition of Example 2

(13) Food Composition

(14) Initially, the core of the product is the composition of Example 2. The amount of which varies in the product from 0.1% to 30% by weight of the product. The introduction of less than 0.1% does not allow the composition to show its antiviral and immunomodulating properties, more than 30% of the activity of the product does not increase, which is not rational.

(15) The composition of the invention may be a food composition, a bioactive food supplement, or a pharmaceutical composition. Food composition means any type of product, drink or confectionery. The food composition, for example, may be a beverage, briquetted cereal concentrate, chewing gum, chocolate, or a dairy product such as a dairy product made by fermentation of a plant product.

(16) Also, the proposed composition can be introduced into cold and hot drinks, sweet foam in aerosol cans, ice cream, cookies, yogurts, cakes, sweets, kielbasa, sausages, dry mixes for feeding children, beer drinks and low alcohol drinks, toothpastes, creams. “Bioactive food supplement” means a food product whose purpose is to supplement a normal diet.

(17) A bioactive nutritional supplement is a concentrated source of nutrients or other substances having a nutritional or physiological effect, alone or in combination. The bioactive nutritional supplement is supplied to the market in a dosage form, namely in such dosage forms as a capsule, lozenge, tablet, pill and other similar forms, sachets with powder, ampoules with liquid, vials equipped with a dropper, and other similar forms of liquid or powder preparations, which are intended for use in measured units of a small amount. The composition may also include amino acids: glycine, arginine, lysine, taurine, phenylalanine or mixtures thereof and any other amino acids.

(18) The composition may also include vitamins: ascorbic acid, thiamine, riboflavin, rutin, folic acid, cyanocobalamin, or a mixture thereof, as well as any other vitamin. The composition may also contain extracts of green or black tea, or any other plant extract used in the food industry. The composition according to the invention, for example, is suitable for daily consumption of cells obtained by culturing a probiotic strain in an amount of from 1-10.sup.8 CFU to 1-10.sup.11 CFU, preferably in an amount of from 1-10.sup.9 CFU to 1-10.sup.10 CFU, when intended for human consumption.

(19) The composition according to the invention, for example, is suitable for daily use of cells obtained by culturing a probiotic strain in an amount of from 1-10.sup.5 CFU to 1-10.sup.11 CFU, preferably in an amount of 1-10.sup.6 CFU to 1-10.sup.10 CFU, when it is intended veterinary applications. The term CFU denotes a colony forming unit.

(20) Probiotics are preferably administered as an oral composition containing metabolically active ones, i.e. probiotic organisms, living and/or lyophilized or non-viable, heat-killed, irradiated or lysed, in the form of spores or covalently chemically modified spores.

(21) The probiotic composition described herein can be administered orally in the form of a tablet, capsule or powder. Additionally, the probiotic composition may be administered orally as a food or nutritious product, such as milk or whey-based fermented milk product or as a pharmaceutical product.

(22) In accordance with one embodiment of the invention, the composition is an edible product, such as a dairy product, a drink, juice, soup or baby food. A probiotic may optionally be combined with at least one suitable prebiotic compound.

(23) Prebiotic “is usually a non-digestible carbohydrate, such as an oligo- or polysaccharide or alcohol sugar, which is not destroyed or absorbed in the upper part of the digestive tract. Known commercially available prebiotics include inulin, fructo-oligosaccharides, oligofructose or galacto-oligosaccharides.

(24) The term “edible product” is intended to encompass all consumed products, especially food products, and may be solid, jelly-like, or liquid. This term covers both finished products and products that are produced by using the probiotic composition as a starter only or in combination with conventional starter cultures or other probiotics. Food products may, for example, be products of the dairy industry or the beverage industry. Alternatively, it may be a natural product.

(25) Initially, the core of the product is the composition of Example 2. The amount of which varies in the product from 0.1% to 30% by weight of the product. The introduction of less than 0.1% does not allow the composition to show its antiviral and immunomodulating properties, more than 30% of the activity of the product does not increase, which is not rational.

(26) In the context of the present invention, “physiologically acceptable diluent or excipent” means any medium or additive that does not affect the effectiveness of the biological activity of the active component (in this case, yeast or bacterial cells), and which is not too toxic to the patient or subject at the administered concentrations.

(27) A physiologically acceptable diluent or excipient may be a diluent or excipient suitable for administration to humans and/or animals (in particular mammals). The pharmaceutical compositions of the present invention may be administered using any effective combination of dosage and route of administration in order to obtain the desired therapeutic effect. Exact amount, intended for administration, may vary from patient to patient, depending on the age, weight and general health of the patient, the origin and severity of diarrhea, etc. The method of administration (oral or parenteral) may be selected depending on the origin of diarrhea and/or depending on the age and/or health of the patient.

(28) By way of example, the invention relates to the pharmaceutical composition as defined above for daily use of cells obtained by culturing said probiotic strain in an amount of 1-108 CFU to 1-10.sup.11 CFU, preferably in an amount of 1-10.sup.9 CFU to 10.sup.10 CFU, in when it is intended for human consumption.

(29) The present invention also relates to the pharmaceutical composition described above, which is intended for daily use of cells obtained by culturing said probiotic strain in an amount of 1-10.sup.5 CFU to 1-10.sup.11 CFU, preferably in an amount of 1-10.sup.6 CFU to 1-10.sup.10 CFU, when it is intended for use in veterinary medicine.

(30) The composition of the pharmaceutical composition according to the present invention may vary depending on the route of administration and dosage in which the composition will be used. After being formulated with at least one physiologically acceptable diluent or excipient, the pharmaceutical composition of the invention may be in any form suitable for administration to a mammal, in particular humans, for example, in the form of lozenges, tablets, sugar coated tablets, capsules, syrup, ointments, injectable solutions, suppositories, etc.

(31) A person skilled in the art knows how to select the most suitable diluents and excipients for preparing this type of composition. Thus, for example, excipients such as water, 2,3-butanediol, Ringer's solution, isotonic sodium chloride solutions, synthetic mono- or diglycerides and oleic acid are often used in the preparation of injectable forms.

(32) Liquid compositions, including emulsions, microemulsions, solutions, suspensions, syrups, elixirs, etc., can be formulated in the presence of solvents, solubilizers, emulsifying agents, oils, fatty acids and other additives such as suspending agents, preservatives, sweeteners, flavoring agents, viscosity-modifying agents, coloring agents, etc.

(33) Solid compositions intended for administration by oral route can be formulated in the presence of an inert excipient such as sodium citrate, and optionally such additives as binders, wetting agents, disintegrants, absorption enhancers, lubricants, etc. In certain embodiments, the pharmaceutical composition of the present invention is formulated to immediately release the active (active) component or active components, in particular yeast cells or bacterial cells obtained by culturing a probiotic strain. Alternatively, a pharmaceutical composition may be provided for the purpose of prolonged or targeted release of the active component or active components or to protect the active component or active components, for example, from intragastric acidity and enzymes. For this purpose, coatings that are resistant to pH and/or gastric enzymes, coatings that are sensitive to pH and/or enzyme activity, or bioadhesive coatings that adhere to the walls of the stomach or intestines, or encapsulation systems can be used.

(34) In addition, the pharmaceutical compositions of the present invention may contain at least one additional active pharmaceutical ingredient (i.e., in addition to yeast or bacterial cells obtained by culturing a probiotic strain). “Active pharmaceutical ingredient” means any compound or substance whose administration has a therapeutic or beneficial effect on the health condition or general condition of the patient or subject to which it is administered.

(35) Thus, the active pharmaceutical ingredient may be active in preventing or treating diarrhea by administering a pharmaceutical composition; may be active against a condition or symptom associated with diarrhea (e.g., fever, vomiting, or abdominal cramps); or may increase the availability and/or activity of the active component or active components of the pharmaceutical composition.

(36) Examples of active pharmaceutical ingredients that may be present in the composition of the present invention include, without limitation, anti-inflammatory drugs, antibiotics, antipyretics, anti-emetics, antihistamines, vitamins, antispasmodics, etc. In an advantageous embodiment, the other active pharmaceutical ingredient is not a Bacillus coagulans strain or cells obtained by culturing such a strain, preferably it is not a bacterial strain or cells obtained by culturing such a strain, more preferably it is not a probiotic strain or cells obtained by culturing such a strain. An example of a pharmaceutical composition according to the present invention is a composition containing yeast or bacterial cells obtained by culturing a probiotic strain as the sole active pharmaceutical ingredient. Unless otherwise defined, all technical and scientific terms used in the description have the same meaning as is generally understood by one of ordinary skill in the art to which the invention pertains. Moreover, all publications, patent applications, all patents and all other references made in the description are included in the description by reference.

(37) In an advantageous embodiment, the other active pharmaceutical ingredient is not a Bacillus coagulans strain or cells obtained by culturing such a strain, preferably it is not a bacterial strain or cells obtained by culturing such a strain, more preferably it is not a probiotic strain or cells obtained by culturing such a strain. An example of a pharmaceutical composition according to the present invention is a composition containing yeast or bacterial cells obtained by culturing a probiotic strain as the sole active pharmaceutical ingredient. Unless otherwise defined, all technical and scientific terms used in the description have the same meaning as is generally understood by one of ordinary skill in the art to which the invention pertains. Moreover, all publications, patent applications, all patents and all other references made in the description are included in the description by reference.

(38) TABLE-US-00003 TABLE 3 Products in which the inclusion of our composition is promising   Additional specific New consumer p/p Product qualities qualities 1. Cold drink Activator of metabolism It prevents intestinal (carbonated, and energy of the body upsets (including non-carbonated) from viral rota- and 2. Hot drink (coffee, tea) Activator of metabolism coronavirus and energy of the body enteritis), protects 3. Briquetted Grain Concentrate against colds 4. Chewing gum Prevents disturbance of the (including flu), microflora of the oral cavity activates the immune and caries system, reduces or 5. Chocolate removes allergic 6. Milk product reactions, improves 7. Fermented milk product the absorption of 8. Sweet foam in aerosol cans basic food 9. Ice cream components 10. Cookies (proteins, 11. Candies carbohydrates, 12. Yoghurts vitamins, fats). It 13. Cakes activates the body's 14. Dry food mixes for children Increases weight gain, metabolism due to prevents bowel disease the stimulation of 15. Dry protein mixes for athletes Increases weight gain, tissue respiration and prevents bowel disease due to the synergism of the components. 16. Sausages of different kind (kielbasa, sardelka, sosiska) 17. Beer drinks 18. Mild alcohol drinks Compensates for immunodeficiency caused by alcohol by the immunomodulatory effect of the composition 19. Toothpastes Prevents tooth decay, activates local immunity in the oral cavity, protects against tonsillitis, scarlet fever, stomatitis (including viral), supports general immunity in tone 20. Cosmetic creams Normalizes skin turgor and local immunity, promotes wound healing after sunburn. 21. Bioadditives (capsule, lozenge, Enhances weight gain (for Components for tablet, pill and other similar athletes and for patients in preventing viral and forms, sachets of powder, the rehabilitation phase), microbial pathologies ampoule with liquid, bottles improves food absorption against the equipped with a dropper) background of immunodeficiency and a means for the comprehensive correction of pathologies caused by impaired immune functions. They can be promising as a Components supplements of sports nutrition with properties to stimulate the body's metabolism and energy (endurance).

(39) Toothpastes and Cosmetic Creams

(40) Initially, the core of the product is the composition of Example 2. The amount of which varies in the product from 0.1% to 30% by weight of the product. The introduction of less than 0.1% does not allow the composition to show its antiviral and immunomodulating properties. However, more than 30% of the activity of the product does not increase, which is not rational. In some embodiments, probiotics release profiles for microorganisms may be controlled, such as, but not limited to, Bacillus coagulans, Bacillus subtilis, Bacillus laterosporus, Bacillus laevolacticus, Sporolactobacillus inulinus, Lactobacus lactobacillus bacillus acidophilus, Lactobacus bacillus bacillus acidophilus Lactobacillus fermentum, Lactococcus lactis, Pedioccocus acidilacti, Pedioccocus pentosaceus, Pedioccocus urinae, Leuconostoc mesenieroides, Bacillus coagulans, Bacillus subtilis, Bacillus laterosporus, Bacillus laevolacticacus, Spor. Breath fresheners are also known under the following trademarks: Retsyn™, Actizol™ and Nutrazin™. Examples of odor control compositions are also described in U.S. Pat. No. 5,300,305 to Stapler et al., And in Patent Publications No. US 2003/0215417 and 2004/0081713, which are incorporated herein by reference for all purposes.

(41) Dental care ingredients (also known as oral care ingredients) may include, but are not limited to, tooth whiteners, stain removers, oral cleansers, whitening agents, desensitizing agents, tooth remineralizing agents, antibacterial agents, anticariotic agents, buffering agents to neutralize plaque acids, surfactants and anti-tartar agents. Non-limiting examples of such ingredients may include hydrolytic agents, including proteolytic enzymes, abrasives such as hydrated silicon dioxide, calcium carbonate, sodium bicarbonate and alumina, other stain removing active ingredients, such as surfactants containing but not limited to anionic surfactants such as sodium stearate, sodium palmitate, sulfonated butyl oleate, sodium oleate, fumaric acid salts, glycerin, hydroxylated lecithin, sodium lauryl sulfate, and chelating agents such as polyphosphates, which are commonly used as anti-calculus ingredients.

(42) In some embodiments, dentifrice ingredients may also include tetrasodium pyrophosphate and sodium tripolyphosphate, sodium bicarbonate, sodium acid pyrophosphate, sodium tripolyphosphate, xylitol, sodium hexametaphosphate. In some embodiments, peroxides are provided, such as carbamide peroxide, calcium peroxide, magnesium peroxide, sodium peroxide, hydrogen peroxide and peroxydisphosphate. In some embodiments, potassium nitrate and potassium citrate are provided.

(43) Other examples may include casein-glycomacropeptide, calcium casein peptone derivative—calcium phosphate, casein phosphopeptides, casein phosphopeptide—amorphous calcium phosphate (CPP-ACP), and amorphous calcium phosphate. Other further examples may include papain, krillase, pepsin, trypsin, lysozyme, dextranase, mutanase, glycoamylase, amylase, glucose oxidase, and combinations thereof.

(44) Additional examples may include surface active substances, such as sodium stearate, sodium ricinoleate and sodium lauryl sulfate, as a surface active substances for use in some embodiments to achieve enhanced prophylactic action, and to make dental care ingredients more acceptable in cosmetic dentistry. Surface active substances may be used preferably as detergents that impart detergent and foaming properties to the composition.

(45) Suitable examples of surfactants are water-soluble salts of higher aliphatic monoglyceride monosulfates monosulfates, such as the sodium salt of monosulfonated fatty acid monoglycerides of hydrogenated coconut oil, higher alkyl sulfates, such as lauryl sulfate sodium alkylaryl sulfonates such as sodium dodecylbenzenesulfonate, higher alkyl sulfoacetates, sodium lauryl sulfoacetate, esters of higher aliphatic acids and 1,2-dihydroxypropanesulfonate, and substantially saturated higher aliphatic acylamides of lower amino aliphatic acids such as having from 12 to 16 carbon atoms in fragments of an aliphatic acid, alkyl or acyl radicals, and the like. Examples of the latter amides mentioned are N-lauroylsarcosine, and the sodium, potassium and ethanolammonium salts of N-lauroyl, N-myristoyl or N-palmitoylsarcosine.

(46) In addition to surfactants, dentifrice ingredients may include antibacterial agents, such but not limited to triclosan, chlorhexidine, zinc citrate, silver nitrate, copper, limonene, and cetylpyridinium chloride. In some embodiments, additional anticariotic agents may include fluoride ions or fluorine-containing components, such as inorganic fluoride salts.

(47) In addition to surfactants, dentifrice ingredients may include antibacterial agents, such but not limited to triclosan, chlorhexidine, zinc citrate, silver nitrate, copper, limonene, and cetylpyridinium chloride. In some discussion, additional anticariotic agents may include fluoride ions or fluorine-containing components, such as inorganic fluoride salts. In some embodiments, a fluorine-containing compound can also be included as an ingredient, which has a beneficial effect on oral care and hygiene, for example, to reduce the solubility of enamel in acid and protect teeth from decay. Examples thereof include sodium fluoride, stannous fluoride, potassium fluoride, potassium stannous fluoride (SnF2-KF), sodium hexafluorostannate, stannous chlorofluoride, sodium fluorozirconate and sodium monofluorophosphate. In some embodiments, urea is included.

(48) Pharmaceutical Compositions

(49) Various methods for preparing a patentable pharmaceutical composition (PFC) can be used in addition to the methods described in Examples 1 and 2.

(50) The PFC composition can be given orally or can be administered by intravascular, subcutaneous, intraperitoneal injection, in the form of an aerosol, by ocular route of administration, into the bladder, topically, and so on. For example, inhalation methods are well known in the art. The dose of the therapeutic composition will vary widely depending on the particular antimicrobial PFC administered, the nature of the disease, frequency of administration, route of administration, clearance of the agent used from the host, and the like. The initial dose may be higher with subsequent lower maintenance doses.

(51) The dose can be administered with a frequency of once a week or once every two weeks, or divided into smaller doses and administered once or several times a day, twice a week, and so on to maintain an effective dose level. In many cases, a higher dose will be needed for oral administration than for intravenous administration. PFCs can be included in many therapeutic compositions. More specifically, the PFCs of the present invention can be incorporated into pharmaceutical compositions in combination with suitable pharmaceutically acceptable carriers or diluents, and can be incorporated into preparations in solid, semi-solid, liquid or gaseous forms, such as capsules, powders, granules, ointments, creams, foams, solutions, suppositories, injections, forms for inhalation applications, gels, microspheres, lotions and aerosols. As such, the administration of the compounds can be carried out in various ways, including oral, buccal, rectal, parenteral, intraperitoneal, intradermal, transdermal, intratracheal administration and so on.

(52) The PFCs of the invention can be distributed systemically after administration or can be localized using an implant or other composition that holds the active dose at the site of implantation. The PFCs of the present invention can be administered alone, in combination with each other, or they can be used in combination with other known compounds (e.g. clopidogrel, anti-inflammatory agents, and so on).

(53) In pharmaceutical dosage forms, the compounds may be administered in the form of their pharmaceutically acceptable salts. The following methods and excipients are given by way of example only and are not in any way limiting. For preparations for oral administration, our compositions can be used alone or in combination with suitable additives for the manufacture of tablets, powders, granules or capsules, for example, with conventional additives such as lactose, mannitol, corn starch or potato starch; with binding agents such as crystalline cellulose, cellulose derivatives, gum arabic, corn starch or gelatins; with disintegrants such as corn starch, potato starch or sodium carboxymethyl cellulose; with lubricants, such as talc or magnesium stearate; and, if desired, with diluents, buffering agents, wetting agents, preservatives and flavoring agents. PFC should be included in the composition for injection by dissolving, suspending or emulsifying them in an aqueous or non-aqueous solvent such as vegetable or other similar oils synthetic aliphatic acid glycerides, esters of higher aliphatic acids or propylene glycol; and, if desired, with conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers and preservatives.

(54) PFCs can be used in an aerosol composition for inhalation administration. The compounds of the present invention can be incorporated into suitable pressure propellants such as dichlorodifluoromethane, propane, nitrogen and the like. In addition, PFCs can be incorporated into suppositories by mixing with a variety of bases, such as emulsifying bases or water-soluble bases. The compounds of the present invention can be administered rectally using a suppository.

(55) A suppository may contain excipients, such as cocoa butter, carboax, and polyethylene glycols, which melt at body temperature but are solid at room temperature. Standard dosage forms for oral or rectal administration, such as syrups, elixirs and suspensions, where each unit dose, for example, a teaspoon, tablespoon, tablet or suppository, can contain a predetermined amount of a composition containing one or more components of the composition of the present invention.

(56) Similarly, unit dosage forms for injection or intravenous administration may contain mixtures of the components of the present invention in a composition in the form of a solution in sterile water, normal saline, or another pharmaceutically acceptable carrier. Implants for sustained release of compositions are well known in this type scientific technique. Implants are made in the form of microspheres, plates, and possible other options, with biodegradable or non-biodegradable polymers.

(57) For example, lactic and/or glycolic acid polymers form a degradable polymer that is well tolerated by the host. An implant containing the PFC according to the invention is positioned close to the focus of the pathology, so that the local concentration of the active agent is increased compared to other areas of the body. As used herein, the term “unit dosage form” refers to physically discrete units suitable for use as single doses for human and animal subjects. Each unit containing a predetermined amount of compounds of the present invention, which, according to calculations, is sufficient to provide the desired effect, together with a pharmaceutically acceptable diluent, carrier or excipient.

(58) The descriptions of the unit dosage forms of the present invention depend on the particular mixture used, the effect to be achieved, and the pharmacodynamics of the mixture used in the host. Pharmaceutically acceptable excipients, such as excipients, adjuvants, carriers or diluents, are generally available. In addition, pharmaceutically acceptable excipients are generally available, such as pH adjusting agents and buffering agents, tonicity agents, stabilizers, wetting agents and the like. Typical doses for systemic administration range from 0.1 pg to 1000 milligrams per kg of subject body weight per administration. A typical dose may be one tablet for administration from two to six times a day, or one capsule or sustained-release tablet for administration once a day with a proportionally higher content of the active ingredient.

(59) The effect of prolonged release may be due to the materials of which the capsule is made, which dissolve at different pH values, capsules that provide a slow release under the influence of osmotic pressure or by any other known controlled release method, including based on covalent modification of bacterial spores. Those skilled in the art will appreciate that dose levels may vary depending on the particular structure or mixture, the severity of the symptoms, and the subject's predisposition to side effects. Some of the specific structures and compositions are more potent than others. Preferred doses of this composition can be readily determined by those skilled in the art in a variety of ways.

(60) The preferred method is to measure the physiological activity of PFC. One of the methods of interest is the use of liposomes as a vehicle for delivery. Liposomes fuse with the cells of the target region and ensure the delivery of liposome contents into the cells. The contact of the liposomes with the cells is maintained for a time sufficient for fusion using various methods of maintaining contact, such as isolation, binding agents and the like.

(61) In one aspect of the invention, liposomes are designed to produce an aerosol for pulmonary administration. Liposomes can be made with purified proteins or peptides that mediate membrane fusion, such as Sendai virus or influenza virus and so on. Lipids can be in any useful combination of known liposome forming lipids, including cationic or zwitterionic lipids, such as phosphatidylcholine.

(62) The remaining lipids will usually be neutral or acidic lipids, such as cholesterol, phosphatidylserine, phosphatidylglycerol and the like. To obtain liposomes, the method described by Kato et al. (1991) J. Biol. Chem. 266: 3361 is utilized. Generally speaking, lipids and a composition for incorporation into liposomes containing peptides are mixed in a suitable aqueous medium, in a suitably salty medium, where the total solids content will be in the range of about 110 wt. %. After vigorous stirring for short periods of approximately 5-60 seconds, the tube is placed in a warm water bath at approximately 25-40° C. and this cycle is repeated approximately 5-10 times. The composition is then sonicated for a suitable period of time, typically approximately 1-10 seconds, and optionally further mixed with a vortex mixer. Then the volume is increased by adding an aqueous medium, usually increasing the volume by about 1-2 times, followed by shaking and cooling. The method allows to include supramolecular structures with high total molecular weight, such as modified microbial spores, in liposomes.

(63) Compositions with Other Active Agents

(64) For use in the methods under consideration, the PFCs of the invention can be formulated with other pharmaceutically active agents, in particular other antimicrobial and antiviral agents. Other agents of interest also include a wide range of antianginal, antihypertensive, thrombolytic, anti-inflammatory agents known in the art. Classes of drugs for the treatment of viral and microbial diarrhea, influenza and its complications are presented in standardized treatment protocols for this pathology and can be combined with patented PFC. Cytokines, such as interferon gamma, tumor necrosis factor alpha, interleukin 12, and so on, may also be included in the PFC composition of the invention. The present invention is further described by the following examples, which should not be construed as limiting the scope of the invention.

Example 4. Protective Antiviral Activity of the Composition Obtained in Example 2, for Example, the Prevention of Mortality in Mice Infected with the H1N1 Influenza Virus (California 2011)

(65) Tests were carried out on 10 groups of young white laboratory mice, 10 in each group. To create immunodeficiency, all animals were injected intraperitoneally with cyclophosphamide once at a dose of 100 mg/kg 2 times a week: in group A, animals were orally given 0.2 ml of water instead of the CMSB composition daily 1 time per day for 10 days. This group acted as a positive control. The second group was administered KCMSB (option with S. boulardii) at a dose of 0.2 ml per animal orally. The third to sixth groups introduced different variants of the KCMSB. The seventh group was administered Lactovit forte (BK with vitamins), the eighth group was administered Enterol (S. boulardii), the ninth group took KCMSB without S. boulardii, without benzyl isoquinolines and without germination activators (gibberellin and others). The tenth group orally received the standard antiviral drug remantadine at a dose of 75 mg/kg daily 1 time per day. After a week of taking cyclophosphamide, the animals developed immunodeficiency at the cellular link. After that, all animals were infected by inoculation of the H1N1 influenza virus (California 2011) at a dose of 10 ED90/mouse intranasally in 0.1 ml. The main criterion for the effectiveness of an anti-influenza drug is the mortality rate of animals after inoculation of the virus. In control group No. 9, the mortality rate was 90%. The research results are presented in table 4.

(66) TABLE-US-00004 TABLE 4 The results of a study of the preventive antiviral effect against the H1N1 influenza virus of various compositions based on KCMSB The Percentage number of of surviving surviving animals animals The name and composition after the after Group of different experiment, experiment, No. variants n = 10 % 1 Control-Normal Saline 1 10 2 KCMSB (variant with S. Boulardii 10 100 and poppy seed extract) 3 KCMSB (papaverine variant) 10 100 4 KCMSB (variant of poppy seed 9 90 extract)) 5 KCMSB (Maca variant, 9 90 S. boulardii, gibberellin, carnitine) 6 All components of 3 30 KHMSB without S. Boulardii and without modified spores of B. coagulans, without probiotics, only plant extracts, papaverine and plant growth activators) 7 Lactovit Forte 3 30 8 Enterol 3 30 9 KCMSB (modified disputes only) 10 Rimantadine 5 50 P < 0.01, the difference is significant for all groups 2-10 against the control of group 1

(67) As can be seen from table 4, the most effective compositions were variants of the invention, containing in one case papaverine and in another papaverine and S. boulardii. In both cases, all infected animals survived. Close results with 90% survival were observed in groups of 4-5 different embodiments of the compositions. A composition that does not include probiotics (including modified spores) had a weak prophylactic anti-influenza activity (Group 6) and protected only 30% of the animals from death. At the same time, the pure spores of S. boulardii from the Lactovit-Forte preparation had significantly less prophylactic antiviral activity (only 30% of the animals survived). The same picture was observed when using Enterol containing S. boulardii. The anti-influenza drug Remantadin was able to protect only 50% of the animals. Thus, for the manifestation of the maximum preventive antiviral activity against the background of chemical immunodeficiency, it is necessary to use the composition of KCMSB with all components of the prescription. Without probiotic components, in particular modified BC spores, the composition loses antiviral prophylactic activity. The presence of S. boulardii in the composition also prevented the death of animals in general. The mechanisms of this prophylactic effect are still not clear.

Example 5. Protective Antiviral Activity of KCMSB Against Rotaviruses

(68) Rotavirus infection is an infectious disease caused by rotaviruses. It is the most common cause of diarrhea in children. This disease is characterized by an acute onset, mild symptoms of gastroenteritis or enteritis, a frequent combination of intestinal and respiratory syndromes in the initial period of the disease. Rotavirus infection is often incorrectly called intestinal flu, although rotavirus has nothing to do with influenza viruses.

(69) In the experiment, the rotavirus subserotype strain IMI B117 was used (Kharkov, Ukraine). This strain causes typical diarrhea in rats with loose stools, an increase in body temperature, a change in behavior (decreased mobility), a drop in appetite, and a deterioration in the appearance of hair (they become ruffled). To fix these characteristics, each rat was placed in a small cage, the bottom of which was covered with white paper. The degree of diarrhea was fixed by the degree of contamination of the paper (in points from 1 to 10). The time of onset of signs of viral diarrhea was also recorded. The first stage of the experiment was the creation of an immunodeficiency background to increase the percentage of animals with viral diarrhea. For this, each animal was injected twice a week with 100 mg/kg of cyclospamide intravenously. Such a dose is able to suppress the cellular component of immunity and block the initial antiviral response of the body. After this week, the animals were given different HCBS oral formulations once a week, once a day, at a dose of 0.5 ml (0.2 g dry composition). Each composition was given to one group of rats, in each group of rats there were 6 animals. There were 10 groups in total. Table 5 presents the distribution of groups and the results of a study of the prophylactic antidiarrheal effect caused by rotavirus B. Each animal was injected with 5 ED90 of the virus in a culture medium in milk (1:1) orally.

(70) TABLE-US-00005 TABLE 5 The results of a study of the preventive antiviral effect against rotavirus B of different compositions based on KCMSB The number of The time animals with that viral % Group The name and composition of diarrhea (8-10 points diarrhea efficiency No. the solution *), (%), n = 6 lasted, days ** 1. Control is Normal Saline 6 9 — 2. Cycloferon 5 (17) 7 (22) 30 3. KCMSB (variant with S. 0 0 100 boulardii and poppy seed extract) 4. KCMSB (papaverine variant) 0 0 100 5. KCMSB (variant of poppy 0 0 100 seed extract)) 6. KCMSB (option poppy 0 0 100 extract, S. boulardii, gibberellin, carnitine) 7. All KCMSB without S. 5 (17) 3 (67) 75 Boulardii and without modified spores of B. coagulans, without probiotics, only plant extracts, papaverine and plant growth activators) 8. Lactovit Forte 3 (50) 5 (44) 72 9. Enterol 3 (50) 5 (44) 72 10. KCMSB (modified disputes 3 (50) 5 (44) 72 only) * 1 point-single spots on paper in cells, 10 points-massive diarrhea with continuous contamination of the paper litter, as well as a set of clinical signs of viral diarrhea. ** percentage of efficiency-the arithmetic average of the sum of the percent of sick animals and the percentage of reduction in the duration of the disease in relation to the control. P < 0.01, the difference is significant for all groups 2-10 against the control of group 1 (Chi-square method)

(71) As can be seen from table 5, none of the animals that received KCMSB containing covalently modified spores of BC together with benzoquinoline alkaloids, with plant extracts and activators of germination of spores did not get sick with viral enteritis and did not show any signs of diarrhea or signs of viral infection. The KCMSB component without probiotics was less effective and accounted for 75% of efficiency, while Lactovit-Forte and Enterol preparations showed only 72% of effectiveness. The least effective was cycloferon (30%), administered orally at a dose of 30 mg/kg body weight every other day. Thus, there is a synergistic activity of all the components included in the composition. The exclusion from the composition of either covalently chemically modified spores of BC or other components of the composition leads to a loss of effectiveness. In addition, attention should be paid to the higher degree of protection of animals with KCMSB compositions against Lactovit-Forte pharmaceutical preparations (BC and vitamins B12, B9) and Enterol (S. boulardii).

Example 6. Protective Antiviral Activity of KCMSB Against Coronaviruses

(72) Respiratory coronavirus is represented by strains OC38, OC43, and the cause of enteritis is most often strain 229 E. The name is explained by the presence of a special ring on the surface of the viral particle: peplomers in the form of club-shaped spike-like outgrowths resembling a corona are present on the lipid membrane or supercapsid. Coronavirus can provoke damage to: the respiratory system; gastrointestinal tract; nervous system. The primary reproduction in the mucous membrane of the nasopharynx and respiratory tract, as a result, there is a profuse runny nose, and in children—bronchitis and pneumonia. In most cases, people encounter diarrhea and enteritis caused by coronaviruses

(73) Alphacoronavirus 1 strain IMI K105 (Kharkov, Ukraine) was used in the experiment. This strain causes typical diarrhea in rats with loose stools, an increase in body temperature, a change in behavior (decreased mobility), a drop in appetite, and a deterioration in the appearance of hair (they become ruffled). To fix these characteristics, each rat was placed in a small cage, the bottom of which was covered with white paper. The degree of diarrhea was fixed by the degree of contamination of the paper (in points from 1 to 10). The time of onset of signs of viral diarrhea was also recorded. The first stage of the experiment was the creation of an immunodeficiency background to increase the percentage of animals with viral diarrhea. For this, each animal was injected twice a week with 100 mg/kg of cyclophosphamide intravenously. Such a dose is able to suppress the cellular component of immunity and block the initial antiviral response of the body. After this week, the animals were given different HCBS formulations orally once a week, at a dose of 0.5 ml (0.2 g dry composition). Each composition was given to one group of rats, in each group of rats there were 6 animals. There were 10 groups in total. Table 6 presents the distribution of groups and the results of a study of the prophylactic antidiarrheal effect caused by coronavirus. Each animal was administered 1 ED90 of the virus in a culture medium orally

(74) TABLE-US-00006 TABLE 6 Results of preventive antiviral action study for coronaviruses of different compositions based on KCMSB The number of The time animals with that viral % Group The name and composition of diarrhea (8-10 diarrhea efficiency No. the components points *), (%), n = 6 lasted, days ** 1. Control is physical. Solution 6 9 — 2. Cycloferon 5 (17) 7 (22) 30 3. KCMSB (version with S. 0 0 100 Boulardii and poppy seed extract) 4. KCMSB (papaverine variant) 0 0 100 5. KCMSB (variant of poppy 0 0 100 seed extract)) 6. KCMSB (variant of poppy, S. 0 0 100 boulardii, gibberellin, carnitine) 7. All KCMSB components 4 (33) 4 (55) 71 without S. Boulardii and without modified spores of B. coagulans, without probiotics, only plant extracts, papaverine and plant growth activators) 8. Lactovit Forte 2 (67) 5 (44) 55 9. Enterol 2 (67) 5 (44) 55 10. KCMSB (modified disputes 2 (67) 5 (44) 55 only) * 1 point-single spots on paper in cells. 10 points-massive diarrhea with continuous contamination of the paper litter, as well as a set of clinical signs of viral diarrhea. ** percentage of efficiency-the arithmetic average of the sum of the percent of sick animals and the percentage of reduction in the duration of the disease in relation to the control. P < 0.01, the difference is significant for all groups 2-10 against the control of group 1 (Chi-square method)

(75) As can be seen from table 6, none of the animals that received KCMSB containing covalently modified spores of BC together with benzoquinoline alkaloids, plant extracts and activators of germination of spores did not get sick with viral enteritis caused by coronavirus, and did not show any signs of diarrhea or viral infection. The KCMSB component without probiotics was less effective and accounted for 75% of efficiency, while Lactovit-Forte and Enterol preparations showed only 71% of effectiveness. The least effective was cycloferon (30%), administered orally at a dose of 30 mg/kg body weight every other day. Thus, there is a synergistic activity of all the components included in the composition. The exclusion from the composition of either covalently chemically modified spores of BC or other components of the composition leads to a significant decrease in the effectiveness of prophylaxis. In addition, attention should be paid to the higher degree of protection of animals with KCMSB compositions against Lactovit-Forte pharmaceutical preparations (BC and vitamins B12, B9) and Enterol (S. Boulardii).

Example 7. Immunomodulatory Activity of KCMSB in Comparison with the Drug Lactovit-Forte

(76) To determine the effect of KCMSB preparations on the immunity of mice, animals were grouped according to the scheme described in example 3, except for the group with remantadine. There were 9 groups in total. Cyclophosphamide (CFA) was administered to all animals once at a dose of 250 mg/kg to induce immunodeficiency as shown in Example 3. After a week, they began to give samples of KCMSB orally at a dose of 0.2 ml (the dose contained 0.1 dry composition KCMSB).

(77) Hsp70, NO, and Cytokine Synthesis Assay.

(78) Isolation of murine peritoneal macrophages and spleen T-lymphocytes, measurement of TNF production was performed by enzyme-linked immunosorbent assay (ELISA) using the ELISA Kit (Mouse TNF-a ELISA Kit, PharMingen, BD Biosciences, San Jose, Calif., USA). NO Macrophages, 106 cells/ml, pH indicator with methylene red at 37° C. in a humidified atmosphere containing 5% CO2. The medium was enriched with 25 mM HEPES, 3% fetal bovine serum. Supernatants were collected after 21 hours and analyzed for N02. Griss reagent (containing 1% sulfanilamide/0.1% naphthylethylenediamine dihydrochloride and 2.5% H3PO4 (1:1)) used to evaluate N02, which served as an indicator of NO formation. After cultivation, 100 ml of each sample was placed in a 96-well plate with 100 ml of Griss reagent. Ten minutes later, the optical density was measured on a tablet spectrophotometer (Stat Fax 303 plus, Awareness technology inc., USA) at 590 nm.

(79) Determination of the Ability of Lymphocytes to Express IL-2

(80) Spleen lymphocytes, 1.5×106 cells/ml, were cultured for 72 hours in 24-well plates in 199 medium at 37° C. in a humidified atmosphere containing 5% CO2. The medium was supplemented with 25 mM HEPES, 5% fetal bovine serum and 5 mg/ml phytohemagglutinin (PHA). The concentration of IL-2 in the supernatant of cells stimulated PHA was measured by ELISA using polyclonal antibodies from rabbits against mouse IL-2 (BioRad, AbD Serotec, Raleigh, N.C., USA) and goat anti-rabbit IgG conjugated with Biotin (Agrisera Antibodies, Vannas, SWEDEN). After ABTS staining, the optical density was measured on a tablet spectrophotometer (Stat Fax 303 plus, Awareness technology inc., USA) at 405 nm. Statistical significance was calculated by t-student test in the modification of Bonferroni. Table 7 shows the results of the effects of KCMSB on mouse immunity.

(81) TABLE-US-00007 TABLE 7 The effect of KCMSB in comparison with the drug Lactovit-Forte on the mice immunity. Cells, tissues, lymphokines NO production Number of Il-2 products (ng/ml) T in mouse macrophages Number of Splenic T Blood Macrophages cells macrophages (in splenocytes Groups lymphocytes plasma (TNF-a,) (TNF-b) μg/ml millions) (million) Control healthy  76 ± 7   164 ± 10  16 ± 3  13 ± 2  10 ± 3  2.0 ± 0.1 126 ± 22 mice Immunodeficiency 12* ± 5*   32* ± 10* 3* ± 1* 8* ± 2  6* ± 2* 1.0* ± 0.1*  87* ± 12* Control after Cyclophosphamide KCMSB (option 87* ± 10* 178* ± 12  19 ± 5  14 ± 2  12 ± 4  1.3 ± 0.2 133 ± 20 with S. Boulardii and poppy seed extract) KCMSB (option 85* ± 10* 177* ± 12  18 ± 5  14 ± 2  13 ± 5  1.4 ± 0.3 135 ± 23 papaverine) KCMSB (variant 89* ± 11* 177* ± 14  19 ± 5  16 ± 3  12 ± 4  1.3 ± 0.2 129 ± 20 of poppy seed extract)) KCMSB (variant 87* ± 10* 179* ± 13  19 ± 5  16 ± 3  13 ± 5  1.4 ± 0.3 137 ± 23 of poppy, S. boulardii, gibberellin, carnitine) All components of 50* ± 10  100* ± 10  7* ± 4*  8 ± 2* 10 ± 4  1.2 ± 0.1 106 ± 15 KCMSB without S. Boulardii and without modified spores of B. coagulans, without probiotics, only plant extracts, papaverine and plant growth activators) Lactovit Forte 67* ± 9*  100* ± 9  12 ± 2  10 ± 1  10 ± 1  1.0 ± 0.1  90 ± 10 Enterol 69* ± 9*   98* ± 9  10 ± 2   9 ± 1  12 ± 1  1.2 ± 0.1  95 ± 12 KCMSB 68* ± 8  100* ± 10   8 ± 4*  9 ± 1* 11 ± 1  1.1 ± 0.1 100 ± 14 (modified spores only) *Statistically significantly different from both controls, P < 0.05.

(82) As can be seen from table 7, the best result of the restoration of immunity to almost normal levels was observed in all experimental groups (3-6), where high-grade KCMSB compositions were used against the background of CFA—immunodeficiency. Moreover, the level of splenic T lymphocytes was statistically significantly higher than in the control. The only inexplicable indicator that goes beyond the concept of immunosuppression is the increase in IL-2 production by splenocytes above the control in group 5, where induced immunosuppression was observed. We connect this fact with uneven effect of CFA on different parts of the immune system and their non-synchronous stimulation by the spore form of B. coagulans. Unlike the composition of group 3, group 7 (without probiotics), although it stimulated immunity against control group 2, did not even restore it to the level of control group 1. Lactovit-Forte and Enterol restored immunity almost to the norm, similarly to chemically modified spores of CD. Higher indicators of the effect on the immunity of KCMSB compositions No. 3-5 are explained by the synergistic action of the components: stimulation germination of spores with gibberellin and other activators, stimulation of division of microbial cells and fungi by papaverine and/or poppy extract, effect on the immunity of plant extracts of licorice, stevia and spirulina.

(83) The Main Advantages of Our Composition:

(84) 1. A powerful preventive effect against viral infections (influenza, coronavirus, rotavirus) comparable to the action of specific vaccines

(85) 2. Strong immunomodulatory effect

(86) 3. The rapid onset of the effect of activation of immunity after 8 hours (for unmodified spores of Bacillus coagulans 72 hours)

(87) 4. The possibility of introducing our composition into the composition of carbonated drinks, iced tea, ice cream, sweet foam in aerosol cans, creams, toothpastes, cosmetics. Modification of the surface of spores leads to an increase in their stability in the environment and to preservatives.

(88) 5. The ability to protect the product containing Bacillus coagulans spores from souring and spoilage for a long time, and the sour product will contain beneficial Bacillus coagulans bacteria that will not harm the body.

(89) 6. Products based on the proposed composition are able to stimulate the metabolism and energy of the body, which is very important in sports and during rehabilitation after serious illnesses, it can be useful in sports nutrition. This effect is caused by the synergism of spores and extracts: spirulina, stevia, alfalfa. For alfalfa, an anabolic effect and real benefits in sports training have been proven. These extracts are not included in the list of doping.