Composition for preventing adhesion of pathogenic microorganisms to human integuments

Abstract

The inventions relate to the field of hygiene and cosmetology, and more particularly to a composition for preventing the adhesion of pathogenic microorganisms to human body cells, comprising a mixture of glycerin, a bromelain enzyme, a cellulase enzyme, and a structuring polymer, and the use of said composition for preparing a cosmetic and / or hygiene product.

Description

[0001] A COMPOSITION THAT PREVENTS ADHESION OF PATHOGENIC MICROORGANISMS TO HUMAN INTEGUMENTS, AND COMPOSITIONS BASED THEREON

[0002] Field of technology to which the invention relates

[0003] The invention relates to hygiene and cosmetology. More specifically, the invention relates to a composition comprising a mixture of glycerin, bromelain, cellulase, and a structure-forming polymer that prevents the adhesion of pathogenic microorganisms to human skin, either alone or when added to hygiene or cosmetic products. The composition also helps stabilize the consumer properties of products and significantly extend their shelf life.

[0004] State of the art

[0005] Adhesion is the most important and essential factor initiating the infectious process (IP). Without adhesion, IP does not develop. All bacteria have a pronounced ability to adhere to organic and inorganic surfaces. For infections that begin in the mucous membranes, overcoming colonization resistance is the first and most essential factor. Preventing microbial adhesion to susceptible host cells is essential for blocking IP at an early stage of its development, which is taken into account when developing prophylactic medications.

[0006] Pathogenic microbes are capable of producing virulence factors (pathogenicity) that facilitate their colonization of the host organism, damaging and destroying its cells. Furthermore, these compounds help the infectious agent evade the host's defense mechanisms.

[0007] Adhesins facilitate bacterial colonization of host tissues. Adhesion molecules ensure the attachment of microbial cells to eukaryotic cells, which is necessary for the penetration of pathogenic bacteria into host tissues.

[0008] Adhesins are specialized macromolecular complexes of microbial cells, including bacterial fimbriae or cell wall surface structures that facilitate the fixation and attachment of pathogens to specific surfaces. Adherent bacteria form cooperative structures with other bacteria much more readily than free bacteria. These communities offer ample opportunity for plasmid exchange. When the number of adherent cells is high, their surfaces form biofilms. Biofilm formation is a multistage process involving cell adhesion, establishment of adhesive contacts, and the development of a multilayered structure within the glycocalyx. Bacteria surrounded by the glycocalyx are inaccessible to immune effects. Bacterial cells periodically leave the biofilm, creating the possibility of new foci of infection (N.V. Seregina, T.V. Chestnova, V.A. Zherebtsova, V.A. Khromushin, "Review of the biophysical characteristics of microbial adhesion")., Bulletin of new medical technologies - 2008 - T. XV, No. 3 - p. 175).

[0009] Bacterial infection often begins with nonspecific cell aggregation based on hydrophobic interactions between bacterial and host cell structures. In contrast, adhesion is a highly specific binding. Some bacterial adhesins are specific not only to the host but also to tissue, and direct the infecting agent to a specific site within the host organism. Most bacterial adhesins are proteins that recognize hydrocarbon receptor structures on eukaryotic cells, such as epithelial cells, phagocytic cells, or erythrocytes. In addition, bacterial adhesins are capable of binding to extracellular matrix glycoproteins such as fibrolactin, laminin, or integrins (Modern Microbiology: Prokaryotes: in 2 volumes: Vol. 2. Translated from English. / Ed. by J. Lengeler, G. Drews, G. Schlegel. - Moscow: Mir, 2005).

[0010] Acne is a multifactorial, polyetiologic disease of the sebaceous glands that affects almost all adolescents to a greater or lesser degree. Adults also face the problem. Various breakouts associated with this condition periodically appear in 40% of men and 54% of women over 25 years of age. In 12% of women and 3% of men, the condition becomes persistent and resistant to treatment. The resulting deep scars and pigmentation are difficult to treat, often requiring cosmetic surgery. As acne progresses, closed comedones stop draining sebum and P. acnes waste products, transforming into inflammatory papulopustular lesions or, gradually increasing in size, becoming open comedones. Therefore, closed comedones resemble a ticking time bomb, ready to explode at any time in an inflammatory reaction.Thus, comedones form due to epithelial cell adhesion, decreased desmosomal disintegration, and an imbalance of intercellular lipids, especially ceramides. P. acnes are lipophilic bacteria that grow under strictly anaerobic conditions. They are found in all areas of the skin with numerous sebaceous glands (face, chest, back, etc.), hydrolyzing the triglycerides of sebum secretion. P. acnes peaks during puberty, occurring on a surface area of ​​1 cm. 2 healthy skin contains approximately 600 microorganisms, while in patients with acne vulgaris, per 1 cm 2 The number of P. acnes can reach 85,000. With an excess of P. acnes, for example, in comedones, their number many times exceeds the content of aerobic bacteria (cocci), which contributes to the transformation of comedones into inflammatory elements.

[0011] Unlike most streptoderma and staphyloderma, P. acnes persists in inflammatory lesions for a long time, despite intensive treatment. Literature data indicate that the slower healing of inflammatory lesions can be explained by the presence of inflammation stimulators synthesized by P. acnes, the pronounced resistance of P. acnes to destruction by neutrophils and monocytes, and the persistence of P. acnes (phagocytosed bacteria remain viable for a long time, despite antimicrobial therapy) (Acne in the practice of a dermatocosmetologist. A.V. Mayorova, V.S. Shapovalov, S.N. Akhtyamova. Moscow: OOO "Firma KLAVEL", 2005)

[0012] Dental plaque is an example of a specialized biofilm formed by several types of bacteria through specific cooperation and adhesion to the tooth surface. The tooth surface is a hub for the development of a microbial community comprising representatives of approximately 300 taxa. This habitat is characterized by a constant flow of saliva and an abundant supply of easily degradable substrates, making the oral cavity a complex system, a kind of continuous cultivation. The clean, sterile surface of the tooth in the oral cavity is initially coated with saliva, forming an initial film containing organic polymers such as proteins, glycoproteins, and polysaccharides. These serve as a substrate for bacteria of the genus Streptococcus, typically S. oralis, S. mutans, S. mitis, and S. sangius, which are the first to colonize this surface.The cells of these bacteria carry adhesion molecules on their surface that specifically interact with the surface carbohydrates of other bacteria, enhancing their deposition. Actinomyces naeslundii predominates among secondary colonizers of the tooth surface, and strict anaerobes such as Veillonella atypica and Provotella loescheii can also predominate, with the latter being capable of promoting the adherence of other anaerobes, such as Actinomyces israeli and Capnocytophaga gingivalis. Thick layers of plaque may harbor other strictly anaerobic bacteria, such as Fusobacterium nucleatum. Their presence indicates the development of a pathological process—tooth decay—which occurs primarily as a result of microbial production of lactic acid in the presence of abundant sucrose.Dental plaque is in contact with the oral mucosa, which can be inhabited, along with other microorganisms, by spirochetes (Treponema), which are capable of penetrating the mucosa and developing in it (Modern Microbiology: Prokaryotes: in 2 volumes: Vol. 2. Translated from English / Edited by I. Lengeler, G. Drews, G. Schlegel. - M .: Mir, 2005).

[0013] Anti-adhesion therapy has advantages over traditional antibacterial chemotherapy because bacteria do not develop resistance to anti-adhesion drugs. This is because anti-adhesion drugs only inhibit binding to host cells without affecting the viability of microorganisms. Furthermore, anti-adhesion agents are effective against both antibiotic-sensitive and antibiotic-resistant bacterial strains.

[0014] Thus, anti-adhesive compounds can be useful when used in cosmetic and hygiene products applied to various surfaces of the human body.

[0015] There are certain issues that complicate the use of anti-adhesive agents. It is known that most pathogenic bacteria simultaneously express several different types of adhesins on their surface during infection. The adhesion process, in addition to adhesins, can be determined by other factors, such as the hydrophobicity and lipophilicity of the cell surface, as well as the strength of mechanical interactions (G. G. Kharseeva, A. Yu. Mironov, A. A. Alieva, "Suppression of Bacterial Adhesion: Modern Approaches, Problems, and Prospects," Advances in Modern Biology, 2019, Vol. 139, No. 5, pp. 506-515).

[0016] Disclosure of the essence of the invention

[0017] The aim of the present invention is to create a composition that prevents the adhesion of various pathogenic microorganisms to human skin.

[0018] The task was solved by creating a composition comprising a mixture of glycerin, bromelain, cellulase, and a structure-forming polymer that prevents enzyme aggregation. The resulting composition demonstrates a universal anti-adhesive effect against various pathogenic microorganisms on the human body.

[0019] The composition can be used either independently or as part of a cosmetic and / or hygienic product.

[0020] The created composition allows for the stability of the consumer properties of hygiene and / or cosmetic products by reducing their bacterial contamination, significantly extending their shelf life. Most importantly, the composition helps maintain the stability of opened products whose application requires hand contact. Contamination of the product with hand bacteria reduces its effectiveness and shortens its shelf life.

[0021] Typically, the solution to this problem involves adding large quantities of preservatives, which can have toxic, carcinogenic, and other negative effects on the body.

[0022] The created composition is based on natural enzymes, which help preserve the product's consumer properties and increase its stability without increasing its toxic effects. On the contrary, they possess their own beneficial properties, such as anti-edematous and anti-inflammatory effects, active restoration of damaged skin, and a soothing effect.

[0023] The enzymes bromelain and cellulase, used together in the composition, enhance each other’s effects.

[0024] The composition contains the enzyme bromelain at a concentration of 1-5% by weight. The amount of this enzyme in the composition may be 1, 2, 3, 4, or 5%, or 1-2%, 1-3%, or 1-4% by weight.

[0025] Preferred concentrations are 2-4%, most preferably 3-4% by weight. The bromelain enzyme can have an activity of 800-2500 gelatin units / g. In more detail, bromelain may have an activity of 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400 or 2500 gelatin units / g or 800-1000, 800-1000, 800-1200, 800-1400, 800-1500, 800-1600, 800-1700, 800-1800, 800-1900, 800-2000, 800-2100, 800-2200, 800-2400, 900-1100, 900-1300, 900-1500, 900-1700. 900-1900, 900-2100, 900-2300, 1000-1200, 1000-1400, 1000-1600, 1000-1800, 1000-2000, 1000-2200, 1100-2400, 1100-1300, 1100-1500, 1100-1700, 1100-1900; 1100-2100, 1200-1400, 1200-1600, 1200-1800, 1200-2000, 1200-2000, 1200-2400, 1300-1500, 1300-1700, 1300-1900, 1300-2100, 1300-2300, 1300-2500, 1400-1800, 1400-2000, 1400-2200, 1400-2400, 1500-1900, 1500-2100, 1500-2300, 1600- 2000, 1600-2200, 1600-2400, 1800-2400 gelatin units / g.The composition also contains the enzyme cellulase in a concentration of 0.02-0.5% by weight.

[0026] The amount of cellulase enzyme in the composition may be 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4 or 0.5% by weight or 0.02-0.4%, 0.02-0.3%, 0.02-0.2%, 0.02-0.1%, 0.02-0.09%, 0.02-0.07%, 0.02-0.05%, 0.03-0.5%, 0.03-0.4%, 0.03-0.3%, 0.03-0.2%, 0.03-0.1%, 0.03-0.09%, 0.05-0.4%, 0.05-0.3%, 0.05-0.2%, 0.05-0.5%, 0.07-0.5%, 0.07-0.4%, 0.07-0.3%, 0.07-0.2%, 0.07-0.1%, 0.1-0.5%, 0.1-0.4%, 0.1-0.3%, 0.1-0.2% by weight.

[0027] Preferred concentrations are 0.05-0.3%.

[0028] The enzyme cellulase can have an activity of 1000-10000 U / g. In more detail, the cellulase enzyme can have an activity of 1000-2000, 1000-3000, 1000-4000, 1000-5000, 1000-6000, 1000-7000, 1000-8000, 1000-9000, 2000-3000, 2000-4000, 2000-5000, 2000-6000, 2000-7000, 2000-8000, 2000-9000, 3000-4000, 3000-5000, 3000-6000, 3000-7000, 3000-8000, 3000-9000, 4000-5000, 4000-6000, 4000-7000, 4000-8000, 4000-9000, 5000-6000, 5000-7000, 5000-8000, 5000-9000, 6000-7000, 6000-8000, 6000-9000, 7000-8000, 7000-9000, 8000-9000 units / year.

[0029] Polymers of acrylic acid (polyacrylates) can be used as a structure-forming polymer (preventing sedimentation and promoting enzyme stabilization) in an amount of 0.2-1.5% by weight. The amount of structure-forming polymer can be 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4% or 1.5% by weight or 0.2-1.3%, 0.2-1.1% 0.2-0.9%, 0.2-0.7%, 0.3-1.5%, 0.3-1.4%, 0.3-1.2%, 0.3-1.1%, 0.3-1.0%, 0.3-0.9%, 0.3-0.7%, 0.4-1.5%, 0.4-1.4%, 0, 4-1,3%, 0, 4-1, 2%, 0,4-1, 1%, 0, 4-1,0%, 0,4-0, 9%, 0,5-1, 5%, 0,5-1, 4%, 0,5- 1,3%, 0, 5-1, 2%, 0,5-1, 1%, 0, 5-1,0%, 0, 5-0, 9%, 0,6-1, 5%, 0,6-1, 4%, 0, 6-1, 3%, 0,6-1, 2%, 0,6- 1, 1%, 0, 6-1, 0%, 0, 7-1, 5%, 0, 7-1, 4%, 0, 7-1, 3%, 0,7-1, 2%, 0.8-1.5%, 0.8-1.4%, 0.8-1.3%, 0.9-1.5%, 0.9-1.4%, 1.0-1.5%, 1.0-1.4% by weight.

[0030] Preferred concentrations of structure-forming polymer are 0.3-1.2% by weight.

[0031] Acrylic acid polymers are selected from the group Ultrez 10, Ultrez 21, Carbopol 940, Carbopol 934, Carbopol 941, mARC-06, Arespol.

[0032] The polymer can act as a carrier.

[0033] The composition also contains glycerin up to 100% by weight.

[0034] The ratio of the components in the composition can be selected according to the description. The composition can be used alone or as part of a cosmetic and / or hygiene product. Such products can take the form of a body cream or gel, a wash-off mask for the face, neck, and décolleté, a cleansing gel for intimate hygiene, a cream mask for the body, hands, or feet, a toothpaste (water-based or anhydrous), a non-abrasive mouth gel, or a liquid oral care product.

[0035] The cosmetic product may be a wash-off product.

[0036] In the production of hygienic and / or cosmetic products, acceptable ingredients are used, selected from the group of surfactants, vegetable oils, emulsifiers, humectants, thickening structuring polymers, abrasive additives, flavor additives, preventive (anti-caryosis, mineral) additives, medicinal plant extracts, pH regulators, and preservatives.

[0037] Vegetable oils may be selected from almond oil, sunflower seed oil, olive oil, avocado oil, grape seed oil, apricot oil, peach oil, sesame oil, wheat germ oil, but are not limited to those listed.

[0038] Emulsifiers may be selected from the following: dimethylalkylammonium hectorite, benzyldimethylstearylammonium hectorite and aluminum magnesium silicate treated with distearyldimethylammonium chloride, poly(oxyethylene / oxypropylene) methylpolysiloxane copolymer, polyoxyethylene-methylpolysiloxane copolymer, methylpolysiloxane copolymer with a branched silicone chain, polyoxyethylene-methylpolysiloxane copolymer with a branched alkyl chain, polyoxyethylene-methylpolysiloxane copolymer with a branched alkyl and silicone chain, cross-linked polyoxyethylene-methylpolysiloxane copolymer, cross-linked polyoxyethylene-methylpolysiloxane copolymer containing an alkyl group, branched silicone modified with polyglycerol and branched silicone modified with polyglycerol containing an alkyl group, polyol esters with fatty acids include glycerol fatty acid esters, polyglycerol fatty acid esters, polyoxyethylene glycerol fatty acid esters,sorbitan fatty acid esters and polyoxyethylene sorbitan fatty acid esters,

[0039] Humectants can be selected from the following: sorbitol, xylitol, propylene glycol, pentylene glycol, amino acids, alpha hydroxy acids such as lactic, malic, tartaric, citric, honey, polysaccharides, panthenol, hydrolyzed proteins, urea, phospholipids. Thickening structuring polymers can be selected from the following: xanthan gum, sodium carrageenate, sodium hyaluronate, magnesium alginate, calcium alginate, magnesium stearate, calcium stearate, cetyl alcohol, lecithin, pectin, beeswax, candelilla wax, carnauba wax, orange peel wax, polyvinylpyrrolidone copolymers, copolymers based on N-vinyl lactam.

[0040] Surfactants can be selected from the following: anionic - sodium lauryl sulfate, sodium laureth sulfate, sodium coco-sulfate, sodium lauroyl sarcosinate; cationic - cetyltrimethylammonium bromide, methyl benzethonium and benzalkonium chloride; nonionic - decyl glucoside, lauryl glucoside, polysorbate; amphoteric - cocamidopropyl betaine, sodium cocoamphoacetate and others.

[0041] Abrasive additives may be selected from the following: silicon dioxide, hydroxyapatite, alkali or alkaline earth metal carbonate or bicarbonate, dicalcium phosphate dihydrate, crushed and standardized-size particles of crushed fruit pit shells, nut shells, coal, coffee beans, eggshells, crustacean chitin, hard jojoba wax granules, pumice, etc.

[0042] Flavoring additives can be selected from the following: fruit, berry, flower, confectionery, dairy, citrus, tea, nut, tobacco, vanilla, mint, etc.

[0043] Prophylactic additives can be selected from the following: triclosan, chlorhexidine, cetylpyridinium chloride, zinc citrate, alcohol, fluoride compounds, calcium glycerophosphate, magnesium glycerophosphate, etc.

[0044] pH regulators can be selected from the following: sodium hydroxide, potassium hydroxide, trisodium phosphate, sodium citrate, triethanolamine, trisamine (tri(-aminoethyl)amine, etc.

[0045] Herbal extracts can be selected from the following: sage extract, calendula extract, green tea extract, rosemary extract, thyme extract, coconut extract, walnut extract, echinacea extract, lavender extract, lemon extract, etc.

[0046] Preservatives can be selected from the following: parabens and their salts, sodium benzoate, potassium sorbate, benzoic acid, etc.

[0047] The composition content in hygienic and / or cosmetic products may be 5-50% by weight. For example, 5-10%, 10-20%, 20-30%, 30-40%, 40-50%, 10-30%, 10-40%, 20-40%, 20-50%, 30-50%. Brief description of the drawings

[0048] Fig. 1 Gel not containing composition.

[0049] Fig. 2 Gel containing the claimed composition.

[0050] Implementation of the invention

[0051] The examples given below are not limiting, but are intended to illustrate practical embodiments of the invention.

[0052] Examples:

[0053] Specific variants of hygienic and cosmetic products according to the invention are given.

[0054] Example 1: Wash-off mask for face, neck and décolleté.

[0055] To prepare a wash-off mask for the face, neck, and décolleté, a composition was prepared: the enzyme bromelain with an activity of 1200 gelatin units / g in an amount of 1%, the enzyme cellulase with an activity of 3000 units / g in an amount of 0.2%, the carbomer Carbopol 934 in an amount of 1%, and glycerin to 100%. Next, the wash-off mask was prepared in accordance with Table 1.

[0056] Table 1.

[0057] 1 2 3

[0058] No. Ingredients % % %

[0059] 1 Composition: 6 10 20

[0060] 2 pH buffers Necessary Necessary Necessary quantity to quantity to quantity to obtain obtaining obtaining pH 7 pH 6 pH 5.5

[0061] 4 Extracts of medicinal 2 3 1

[0062] plants

[0063] 5 Water Up to 100 Up to 100 Up to 100

[0064]

[0065] Example 2: Intimate hygiene gel.

[0066] To prepare an intimate hygiene gel, a composition was prepared: the enzyme bromelain with an activity of 2200 gelatin units / g in an amount of 2%, the enzyme cellulase with an activity of 9000 units / g in an amount of 0.3%, carbomer Ultrez 21 in an amount of 1.2%, and glycerin to 100%. Next, the intimate hygiene gel was prepared in accordance with Table 2. Table 2.

[0067] 1 2 3

[0068] No. Ingredients % % %

[0069] 1 Composition: 12 18 30

[0070] 2 Cocamidopropyl betaine 15 7 12

[0071] 3 Sodium cocoamphoacetate ___ 10 10

[0072] 4 Pentylene glycol ___ 1 3 Polysorbate-20 2 1.2 0.8

[0073] 5 Xylitol 1 3 ___

[0074] 6 Extracts of medicinal plants 0.2 1 0.8

[0075] 7 pH - buffers pH 5 pH 6 pH 7

[0076] 8 Preservatives 0.5 0.8 1

[0077] 9 Aromatic compositions 0.2 0.3 0.5

[0078] 10 Water Up to 100 Up to 100 Up to 100

[0079]

[0080] Example 3: Toothpaste.

[0081] To prepare the toothpaste, a composition was prepared: the enzyme bromelain with an activity of 800 gelatin units / g in an amount of 4%, the enzyme cellulase with an activity of 9000 units / g in an amount of 0.05%, carbomer mARC-Ob in an amount of 0.3%, and glycerin up to 100%. Then, the toothpastes were prepared in accordance with Table 3.

[0082] Table 3.

[0083] 1 2 3

[0084] No. Ingredients % % %

[0085] 1 Composition 35 42 49

[0086] 2 Silicon dioxide 16 12 8

[0087] 3 Xylitol 10 8 4

[0088] 4 Xanthan gum 1.5 1.2 1.0

[0089] 5 Sodium alginate 1.5 ___ ___

[0090] 6 Sodium lauryl sulfate 1 1.2 1.4

[0091] 7 Alkylamidopropyl betaine 0.3 0.2 ___

[0092] 8 Polysorbate-20 0.2 ___ 0.5

[0093] 9 Sodium monofluorophosphate 0.8 1.1 ___

[0094] 10 Calcium glycerophosphate ___ 0.2 0.8

[0095]

[0096] And Aromatic composition 0.5 0.8 1.2 12 Preservatives 1.2 1.0 0.8

[0097] 13 pH buffers Necessary Necessary Necessary quantity to quantity to quantity to obtain obtaining obtaining pH 7.5 pH 7 pH 6.5

[0098] 14 Sorbitol 20 16 12

[0099] 15 Water 13.5 16.3 21.3

[0100]

[0101] Preparation of the composition:

[0102] 1. The required amount of glycerin is placed in a container with a stirrer and heated while stirring to a temperature of 60-80°C.

[0103] 2. Slowly add the polymer carbomer to the heated glycerin while stirring and stir until a homogeneous gel is formed.

[0104] 3. Cool the gel to 20-30°C and, while stirring, add the cellulase and bromelain enzymes sequentially. Stir until a homogeneous, opalescent gel forms.

[0105] 4. The composition obtained in this way can be stored for a long time and used for the preparation of cosmetic products.

[0106] Preparation of a cosmetic product containing a composition in the form of a toothpaste:

[0107] 1. Place the required amount of water, xylitol, and sorbitol in a container with a stirrer. While stirring, add preservatives, preventative additives (calcium glycerophosphate or sodium monofluorophosphate), and pH buffers. Heat to a temperature of 40-60°C and stir until a clear or opalescent solution is formed.

[0108] 2. Add sodium alginate and / or xanthan gum and mix until a homogeneous gel is formed.

[0109] 3. Add silicon dioxide to the resulting gel, stir until a homogeneous mass is formed, and reduce the temperature to 30-40°C.

[0110] 4. Add the composition, mix and vacuum until a homogeneous mass is obtained.

[0111] 5. Add the aromatic composition and surfactants (sodium lauryl sulfate, alkylamidopropyl betaine, polysorbate-20), mix until a homogeneous product is obtained.

[0112] Determination of anti-adhesive activity against the test strain Propionibacterium acne / Cutibacterium acnes.

[0113] To evaluate the anti-adhesive activity of the proposed compositions, the following studies were conducted using a human embryonic skin fibroblast cell culture. The human embryonic skin fibroblast cell culture model is versatile and suitable for studying the adhesive properties of microorganisms and their formation of microbial biofilm, as well as for assessing the antitoxic and anti-adhesive effects of antiseptics and other agents. This model allows for the evaluation of the specific activity of specific agents simultaneously against a large number of pathogens on wound surfaces and other surfaces, under conditions as close as possible to those found in the macroorganism.

[0114] Afinogenova, A.G. Microbiological aspects of the development and use of antiseptics and antiseptic agents for the prevention and treatment of wound infections. Abstract of a dissertation for the degree of Doctor of Biological Sciences. Moscow, 2011. Methodological recommendations No. 96 / 247. Testing of drugs for external use in human skin cell culture.

[0115] Example 4:

[0116] 1. To evaluate the anti-adhesive activity of the complex against the test strain Propionibacterium acne / Cutibacterium acnes, formulations 1 and 3 (wash-off mask) were prepared in accordance with the Table of Examples 1. Laboratory samples of formulations 1 and 3 were tested as a 10% solution on Eagle's medium. Test culture: Propionibacterium acne / Cutibacterium acnes, the study was carried out under anaerobic conditions. Cell culture of human embryonic muscle fibroblasts. The cells were co-incubated with solutions of formulations 1 and 3 at various dilutions (from a 10% solution) for 3 minutes and 15 minutes to evaluate the toxicity of the test formulations on the cell culture. With an exposure of 3 minutes, sample dilutions of 0.2% and below were safe for the cells. With an exposure of 15 minutes, sample dilutions of 0.12% and below were safe for the cells.At both exposures, during co-incubation of fibroblasts with Propionibacterium acne / Cutibacterium acnes, almost complete destruction of the monolayer (up to 90%) was observed. The adhesion index was 50 CFU per 1 eukaryotic cell. Co-incubation of fibroblasts with 0.2% dilutions of the compositions did not lead to the destruction of the cell monolayer after an exposure of 3 minutes, with 0.12% after an exposure of 15 minutes. During co-incubation of fibroblasts with the test strain and the appropriate dilution of the compositions, the intensity of the adhesion process of the test strain was assessed by the following parameters: 1) the adhesion index (AI) is expressed as the average number of bacterial cells per one eukaryotic cell, 2) the percentage of affected cells of the monolayer (PI%), 3) the contamination of 100 cells of the monolayer - the microbial load (ML) is determined by the formula ML = IA * PI%. The degree of microbial adhesion is determined by the microbial load indicator relative to the control, taken as 100%.

[0117] 2. When incubating cells with sample composition 1 at a dilution of 0.2% with the test strain Propionibacterium acne / Cutibacterium acnes with an exposure of 3 minutes, a destruction of the fibroblast monolayer of up to 10% was observed, with an adhesion index of 1-2 CFU per 1 eukaryotic cell.

[0118] 3. When incubating cells with sample composition 3 at a dilution of 0.2% with the test strain Propionibacterium acne / Cutibacterium acnes with an exposure of 3 minutes, no destruction of the fibroblast monolayer was observed, nor was there any adhesion of microbial cells to fibroblasts.

[0119] 4. When cells were incubated with sample 1 at a 0.12% dilution with the Propionibacterium acne / Cutibacterium acnes test strain for 15 minutes, 80% of the fibroblast monolayer was disrupted. Thus, the remaining fibroblasts (20%) were infected with the test strain. The adhesion index was 20-25 CFU per eukaryotic cell. The microbial load was 500 CFU.

[0120] 5. When cells were incubated with sample 3 at a 0.12% dilution with the Propionibacterium acne / Cutibacterium acnes test strain for 15 minutes, no more than 10% of the fibroblast monolayer was disrupted. The adhesion index was 1-2 CFU per eukaryotic cell. The majority of microbial cells were located in the intercellular space. The percentage of damaged fibroblasts was 1%. The microbial load was 1 CFU.

[0121] Example 5: Determination of anti-adhesive activity against the test strain Staphylococcus aureus.

[0122] To evaluate the anti-adhesive activity of the complex against the test strain Staphylococcus aureus, formulations 1 and 3 (intimate hygiene gel) were prepared in accordance with the table of examples 2. Laboratory samples of formulations 1 and 3 were tested as a 10% solution on Eagle's medium. Test culture: Staphylococcus aureus ATCC. Human embryonic skin-muscle fibroblast cell culture. The cells were co-incubated with solutions of formulations 1 and 3 at various dilutions (from a 10% solution) for 3 minutes and 40 minutes to evaluate the toxicity of the tested formulations on the cell culture. With an exposure of 3 minutes, sample dilutions of 0.1% and below were safe for the cells. With an exposure of 30 minutes, sample dilutions of 0.06% and below were safe for the cells. With a 3-minute exposure during co-incubation of fibroblasts with S. aureus, minor damage to the monolayer (up to 10%) was noted, but pronounced adhesion of the test strain to the cells was observed.The adhesion index was 100-120 CFU per eukaryotic cell. After a 30-minute exposure, co-incubation of S. aureus with fibroblast culture resulted in 80% cell monolayer disruption. The adhesion index was approximately 60-70 CFU per eukaryotic cell. Co-incubation of fibroblasts with 0.1% dilutions of the compounds did not result in cell monolayer disruption after a 3-minute exposure, and 0.06% after a 30-minute exposure. During co-incubation of fibroblasts with the test strain and the corresponding dilution of the compositions, the intensity of the test strain adhesion process was assessed by the following parameters: 1) the adhesion index (AI) is expressed as the average number of bacterial cells on one eukaryotic cell, 2) the percentage of affected cells of the monolayer (PC%), 3) the contamination of 100 cells of the monolayer - the microbial load (ML) is determined by the formula ML = IA * PC%. The degree of microbial adhesion is determined by the microbial load indicator relative to the control, taken as 100%.

[0123] When incubating cells with a sample of composition 1 in a 0.1% dilution with the test strain S. aureus with an exposure of 3 minutes, a destruction of the monolayer of up to 10% was observed, with an adhesion index of 8-10 CFU per 1 eukaryotic cell.

[0124] When incubating cells with a sample of composition 3 at a dilution of 0.1% with the test strain S. aureus with an exposure of 3 minutes, no destruction of the fibroblast monolayer was observed, as well as no adhesion of microbial cells to fibroblasts.

[0125] When cells were incubated with sample 1 at a 0.06% dilution with the S. aureus test strain for 30 minutes, up to 50% of the fibroblast monolayer was disrupted. The adhesion index was 40-50 CFU per eukaryotic cell. The microbial load was 500 CFU.

[0126] When incubating cells with a sample of composition 3 at a dilution of 0.06% with the test strain S. aureus with an exposure of 30 minutes, a 5-10% destruction of the fibroblast monolayer was observed, as well as a lack of adhesion of microbial cells to fibroblasts.

[0127] The conducted tests demonstrate the universal nature of the excellent anti-adhesive action of the proposed composition in interrupting the adhesion of various pathogenic microorganisms.

[0128] To assess the shelf life of formulations containing the composition, a comparative assessment of the stability of laboratory products and regular products containing the bromelain enzyme (Comparison toothpaste containing silicon dioxide, xylitol, xanthan gum, sodium lauryl sulfate calcium glycerophosphate, 1.9% bromelain by weight, without cellulase and structure-forming polymer) was carried out. The determination of the shelf life of products containing the composition was carried out using the example of toothpaste samples manufactured in accordance with Example 3, Table 3, Sample 3.

[0129] The tests were conducted using the "accelerated aging" method in accordance with the recommendations of OFS.1.1.0009.15 "Shelf Life of Medicines." The "accelerated aging" method involves maintaining the test product (toothpaste) at temperatures and humidity levels exceeding its normal storage conditions. Elevated temperatures typically accelerate the physicochemical processes occurring within the product, leading to undesirable quality changes over time. Therefore, at elevated temperatures, the period of time during which the product's controlled quality parameters remain within acceptable limits (the experimental shelf life) is artificially shortened compared to the shelf life at the storage temperature. This significantly reduces the time required to establish the shelf life.

[0130] Laboratory toothpaste samples were packaged in polyethylene tubes with a protective membrane and placed in a climatic chamber at 60°C for 45, 60, and 74 days, simulating the toothpastes' shelf lives of 3, 4, and 5 years, respectively. After testing, the samples were tested for compliance with the technical requirements for finished products according to GOST 7983-99 "Toothpastes. General Specifications." The test results are presented in Table 4.

[0131] Table 4.

[0132] Show Sample salary in accordance with Sample salary comparison

[0133] ately example 3, table 3, sample 3.

[0134] Expired 45 days. 60 days. 74 days Expired 45 days. 60 days. 74 days.

[0135] External Homogeneous Homogeneous Homogeneous Homogeneous Violation Significant NII one ...

[0136]

[0137] and masses Color Light Light Light Light White Light Brown Dark yellow yellow brown brown yellow brown red red Taste Pleasant Pleasant Pleasant Sweet Pleasant Pleasant Sweet Unpleasant, sweet, sweet, sweet, sweet sweet insignificant sweet sweet note

[0138] aty aty aty of body aty aty of bitterness

[0139] note

[0140] bitterness

[0141] The smell of Mint Mint Mint Mint, Mint Mint Mint Unpleasant, not significantly rancid, medicinal, medicinal notes

[0142] skoy

[0143] note

[0144] pH 6.5 6.5 6.0 5.8 6.5 6.5 6.0 5.4 (20%

[0145] suspe

[0146] Nziya)

[0147]

[0148] Based on the results of this experiment, it can be noted that the shelf life of toothpaste containing the stated composition may be longer than that of regular toothpaste containing the bromelain enzyme, and is at least 5 years, while regular toothpaste can be stored without significant changes in quality for no more than 3 years.

[0149] Example 6: Study of microbial contamination of cosmetic gel.

[0150] An experiment was conducted to demonstrate the difference in microbial contamination between a cosmetic gel containing the claimed composition and a regular gel without the composition. A mixture of preservatives was used: sodium benzoate - 0.5%, potassium sorbate - 0.3%, and methylparaben - 0.15% by weight.

[0151] The following composition was used to prepare the gel: the enzyme bromelain with an activity of 1200 gelatin units / g in an amount of 2% by weight, the enzyme cellulase with an activity of 10,000 units / g in an amount of 0.02% by weight, carbomer Ultrez 21 in an amount of 0.25% by weight, glycerin up to 100%.

[0152] The content of this composition in the gel was 10% by weight.

[0153] The gels were stored in a glass container with a lid at room temperature for two months. The test results are shown in Figures 1 and 2.

[0154] Figure 1 shows the formation of a mold colony, while Figure 2 shows no microbial contamination of the surface.

Claims

CLAUSES OF THE INVENTION 1. A composition that prevents the adhesion of pathogenic microorganisms to human body cells, including a mixture of glycerin, bromelain enzyme, cellulase enzyme and a structure-forming polymer.

2. The composition according to claim 1, wherein the amount of bromelain is 1-5% by weight.

3. The composition according to paragraphs 1-2, in which the activity of the bromelain enzyme is 800-2500 gelatin units / g.

4. The composition according to claim 1, in which the amount of cellulase is 0.02-0.5% by weight.

5. The composition according to claim 4, wherein the activity of the cellulase enzyme is 1000-10000 U / g.

6. The composition according to claim 1, in which the amount of structure-forming polymer is 0.2-1.5% by weight.

7. The composition according to claim 1, wherein the structure-forming polymer is at least one polymer of acrylic acid.

8. The composition according to claim 1, in which glycerin is added up to 100% by weight.

9. Use of the composition according to paragraphs 1-8 for the preparation of a cosmetic and / or hygienic product.

10. Use according to paragraph 9, wherein the product may be in the form of a cream or gel for the body, a wash-off mask for the face, neck and décolleté, a washing gel for intimate hygiene, a cream mask for the body, hands or feet, toothpaste, a non-abrasive gel for the mouth, a liquid mouthwash.

11. The use according to claim 9, wherein the product may be a rinse-off product.

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