Composition for the improvement, prevention, or treatment of vaginitis or vulvitis, containing Lactobacillus salivarius HHuMin-U

Abstract

This invention provides a composition containing Lactobacillus salivarius HHuMin-U (KCCM13001P) for use in improving, preventing, or treating vaginitis or vulvitis. [Solution] A pharmaceutical composition for the prevention or treatment of vaginitis or vulvitis is provided, containing Lactobacillus salivarius HHuMin-U (KCCM13001P) as an active ingredient.

Description

[Technical Field]

[0001] This invention relates to a composition for the improvement, prevention, or treatment of vaginitis or vulvitis, comprising Lactobacillus salivarius HHuMin-U. [Background technology]

[0002] Vaginal and vulvar inflammation is the most common reason women visit a gynecologist. Common and representative types of vaginal and vulvar inflammation include bacterial vaginosis, trichomoniasis, and candidiasis, as well as herpes simplex virus infections that cause vulvar ulcers and vulvar warts.

[0003] Vaginitis is an inflammation of the vaginal mucosa caused by bacterial infection. Common causative bacteria include Neisseria gonorrhoeae, Streptococcus, Staphylococcus, Escherichia coli, Candida, and Trichomanus. It can also occur as a symptom of infectious diseases such as tuberculosis, syphilis, diphtheria, typhoid fever, and measles. Common symptoms include increased vaginal discharge, which is often cloudy white or yellowish-white, sometimes purulent or bloody, and usually has a foul odor. The vulva is constantly moist, itchy, and burning. If the vulva becomes inflamed due to the discharge, urination can be painful. While it can sometimes be cured with antibiotic vaginal suppositories, it is crucial to receive proper treatment under a doctor's supervision for early and complete recovery. Furthermore, maintaining good hygiene is paramount for prevention.

[0004] Vulvitis is an inflammation of the vulva, the external genitalia of women, which are the visible parts of the female reproductive organs. The normally split area is called the vulvar cleft, and it includes the labia majora, labia minora, clitoris, vaginal opening, and urethral opening. The main causes of vulvitis can be divided into infectious and non-infectious causes. The most common infectious cause is bacterial infection, which mostly occurs due to an imbalance in the normal bacterial flora of the vagina and is often associated with bacterial vaginosis. Fungal infections are also common, often caused by the yeast Candida albicans, and are characterized by itching, redness, and abnormal discharge. Herpesvirus and human papillomavirus infections can also induce vulvitis, and sexually transmitted infections such as Chlamydia, Neisseria gonorrhoeae, and Trichomonas are known to cause inflammation of the vulva. Non-infectious vulvitis can be caused by allergic reactions, with chemicals found in cleansers, perfumes, sanitary napkins, and condoms irritating the vulva and inducing inflammation. Hormonal changes are also a significant cause; especially during menopause, estrogen levels decrease, making the vulvar skin thinner and drier, thus increasing sensitivity to irritation. Physical irritation can also be a contributing factor, with tight clothing and friction irritating the vulva.

[0005] The most common symptoms include itching, burning, and vulvar pain, along with redness and swelling of the vulvar skin, and abnormal vaginal discharge.

[0006] Under normal conditions, a woman's vagina maintains a slightly acidic environment due to the many lactic acid bacteria inside, which helps to inhibit the invasion of pathogenic microorganisms and the growth of fungi.

[0007] This slightly acidic environment within the vagina can change when external factors, such as excessive vaginal douching, long-term use of antibiotics or contraceptives, or disruption of hormone balance due to pregnancy, childbirth, or menopause, cause changes in the amount of beneficial bacteria, such as lactic acid bacteria, that normally exist in the vagina.

[0008] While sulfonamides, antibiotics, and chemically synthesized disinfectants can be used to treat vaginitis, the use of these drugs is fatal not only to harmful bacteria but also to beneficial lactic acid bacteria, and in particular, the use of antibiotics can induce antibiotic resistance. Furthermore, chemically synthesized disinfectants (povidone-iodine) may cause hypersensitivity reactions to iodine, as well as itching, burning, and dermatitis due to mucosal damage, and there is a problem that elevated iodine levels can induce hypothyroidism.

[0009] Therefore, there is a need for a method that is highly effective in improving and treating vaginitis and vulvitis, and that does not affect the gut microbiota, thereby preventing secondary vaginitis and vulvitis induced by imbalances in the gut microbiota. [Prior art documents] [Patent Documents]

[0010] [Patent Document 1] Korean Patent No. 10-1860513 [Patent Document 2] Korean Patent No. 10-2337950 [Overview of the project] [Problems that the invention aims to solve]

[0011] The present invention aims to provide a composition containing Lactobacillus salivarius HHuMin-U (KCCM13001P) for use in improving, preventing, or treating vaginitis or vulvitis. [Means for solving the problem]

[0012] The present invention provides a pharmaceutical composition for the prevention or treatment of vaginitis or vulvitis, comprising Lactobacillus salivarius HHuMin-U (KCCM13001P) as an active ingredient.

[0013] At this time, as an example, the strain can be any one or more selected from viable cells of Lactobacillus salivarius HHuMin-U (KCCM13001P) strain, its disrupted product, its culture solution, its culture, its extract, and its dead cells.

[0014] At this time, as an example, the vaginitis or vulvitis can be any one selected from bacterial vaginitis, trichomonas vaginitis, candidal vaginitis, atrophic vaginitis, herpes virus infection, and vulvar warts.

[0015] The present invention provides a health functional food for improving vaginitis or vulvitis containing Lactobacillus salivarius HHuMin-U (KCCM13001P).

[0016] The present invention provides a skin external preparation for improving vaginitis or vulvitis containing Lactobacillus salivarius HHuMin-U (KCCM13001P).

[0017] The present invention provides a cleaning agent composition for improving vaginitis or vulvitis containing Lactobacillus salivarius HHuMin-U (KCCM13001P).

Effects of the Invention

[0018] The composition according to the present invention exhibits excellent antibacterial activity against the causative bacteria of vaginitis and vulvitis, and is very effective in improving and treating vaginitis and external inflammation.

Brief Description of the Drawings

[0019] [Figure 1] It is a diagram showing the results of confirming the antibacterial activity of Lactobacillus salivarius HHuMin-U strain according to the present invention against Candida albicans strain. [Figure 2]This figure shows the results of evaluating the effect of the Lactobacillus salivarius HHuMin-U strain according to the present invention on the adhesion strength of Candida albicans strains to vaginal epithelial cells. [Figure 3] This figure shows the results of evaluating the effect of the Lactobacillus salivarius HHuMin-U strain according to the present invention on the inhibition of vaginal epithelial cell death ability by Candida albicans strains. [Figure 4] This figure shows the results of evaluating the effect of the Lactobacillus salivarius HHuMin-U strain according to the present invention on the growth of Candida albicans strains in vaginal epithelial cells. [Figure 5] This figure shows the results of evaluating the effect of the Lactobacillus salivarius HHuMin-U strain according to the present invention on the expression of antimicrobial peptides in vaginal epithelial cells (upper panel: mRNA, lower panel: protein). [Figure 6] This figure shows the results of evaluating the effect of the Lactobacillus salivarius HHuMin-U strain according to the present invention on the expression of immunoactive substances in vaginal epithelial cells (upper panel: mRNA, lower panel: protein). [Figure 7] This figure shows the results of evaluating the effects of oral administration of the Lactobacillus salivarius HHuMin-U strain according to the present invention on the improvement and treatment of vaginitis (A: change in body weight, B: measurement of C. albicans CFU in vaginal douche, C: PAS staining of vaginal tissue). [Figure 8] This figure shows the results of an evaluation of the effects of vaginal administration of the Lactobacillus salivarius HHuMin-U strain according to the present invention on the improvement and treatment of vaginitis. [Modes for carrying out the invention]

[0020] In the following, in order to avoid clutter with redundant information, we have attempted to omit descriptions of repetitive content. In other words, the content of the invention is not limited to the content described below, but should be interpreted based on the overall content of the invention.

[0021] In one embodiment of the present invention, the Lactobacillus salivarius HHuMin-U (KCCM13001P) strain was found to have excellent antibacterial activity against strains that induce vaginitis and vulvitis, thereby completing the present invention.

[0022] Furthermore, the bacterial strain according to the present invention inhibits the adhesion of bacterial strains that induce vaginitis and vulvitis to human vaginal epithelial cells, reduces the degree of death of human vaginal epithelial cells induced by these bacterial strains, and exhibits the effect of increasing the expression of antimicrobial peptides and immunoactive substances in these cells.

[0023] Furthermore, the bacterial strain according to the present invention effectively killed vaginitis and vulvitis-inducing bacterial strains in animal experiments by oral or vaginal administration, effectively suppressing infection of vaginal tissue by vaginitis and vulvitis-inducing bacterial strains.

[0024] Therefore, the present invention provides a pharmaceutical composition for the prevention or treatment of vaginitis or vulvitis, a health functional food for improving the same, a topical skin preparation, or a cleansing agent composition containing Lactobacillus salivarius HHuMin-U (KCCM13001P) as an active ingredient.

[0025] Here, the aforementioned strain may be one or more selected from, for example, live Lactobacillus salivarius HHuMin-U (KCCM13001P) strain, its crushed product, its culture medium, its culture, its extract, and its dead cells.

[0026] The term "the crushed material" used in this invention refers to the crushed material of a bacterial strain, and specifically refers to a bacterial strain that has been crushed by physical force or chemical treatment.

[0027] In this invention, the term "culture medium" refers to the culture medium in which the bacterial strain has been cultured, and a standard lactic acid bacteria culture medium can be used for culturing the bacterial strain. Furthermore, in this invention, "culture medium" is interpreted to include a concentrated form of the culture medium.

[0028] In this invention, the term "culture" refers to a powder produced by drying or volatilizing the culture solution in which a bacterial strain has been cultured.

[0029] In this invention, the term "extract" refers to the extract obtained by adding an extraction solvent to a bacterial strain.

[0030] In this invention, the term "extract" refers to a powder produced by drying or volatilizing the extraction solvent from an extract obtained by adding an extraction solvent to a bacterial strain.

[0031] The term "dead bacterial cells" as used in this invention refers to bacterial strains that have been killed, and the killing of the bacterial strain means killing by applying heat, applying pressure, grinding with a bead, or by enzymatic and chemical treatment, but it is interpreted to include so-called "dead bacterial fragments" in which the bacterial cells have been crushed by the killing treatment.

[0032] In this case, the vaginitis or vulvitis may be, for example, one of the following selected from bacterial vaginosis, trichomoniasis, candidiasis, atrophic vaginitis, herpes simplex virus infection, and vulvar warts.

[0033] The Lactobacillus salivarius HHuMin-U (KCCM13001P) according to the present invention is a strain disclosed in Korean Patent No. 10-2337950 (December 7, 2021). This strain, Lactobacillus salivarius HHuMIN-U, was internationally deposited by Bifido Company Limited on June 1, 2021, with the Korean Culture Center of Microorganisms (Yurim B / D 45, Hongjenae-2ga-gil, Seodaemun-gu, SEOUL, 03641, Republic of Korea), the international depositary under the Budapest Convention, and was assigned accession number: KCCM13001P.

[0034] As one way to achieve the above objective, the present invention provides a pharmaceutical composition for the prevention or treatment of vaginitis or vulvitis, comprising Lactobacillus salivarius HHuMin-U (KCCM13001P) as an active ingredient.

[0035] As used in this invention, the term "prevention" means all actions that suppress or delay the onset of diseases affected by the administration of the pharmaceutical composition according to the present invention, namely vaginitis and vulvitis.

[0036] Furthermore, the term "treatment" as used in this invention means all actions by which the symptoms of vaginitis and vulvitis are improved or beneficially altered by the administration of the pharmaceutical composition according to the present invention.

[0037] The bacterial strain according to the present invention may contain one or more active ingredients that exhibit the same or similar functions as the above-mentioned components.

[0038] The pharmaceutical composition according to the present invention may further contain a pharmaceutically acceptable carrier in addition to the bacterial strain according to the present invention.

[0039] The type of carrier that can be used in the present invention is not particularly limited, and any carrier commonly used in the art can be used. Non-limiting examples of such carriers include lactose, dextrose, sucrose, sorbitol, mannitol, saline solution, sterile water, Ringer's solution, buffered saline, albumin injection solution, xylitol, erythritol, maltitol, maltodextrin, glycerol, and ethanol. These can be used individually or in combination of two or more.

[0040] Furthermore, the pharmaceutical compositions according to the present invention may be used with the addition of other pharmaceutically acceptable additives such as antioxidants, excipients, diluents, buffers, or bacteriostatic agents, if necessary, and may be used with the additional addition of surfactants, binders, fillers, bulking agents, wetting agents, disintegrants, dispersants, or lubricants.

[0041] In the pharmaceutical composition according to the present invention, the bacterial strain according to the present invention may be included in an amount of 0.00001% to 99.99% by weight, preferably 0.1% to 90% by weight, more preferably 0.1% to 70% by weight, and even more preferably 0.1% to 50% by weight, based on the total weight of the pharmaceutical composition, but is not limited thereto, and can be varied in various ways depending on the condition of the subject, the specific type of disease, the degree of progression, etc. If necessary, it may also be included in the total content of the pharmaceutical composition.

[0042] In other words, the pharmaceutically effective amount and effective dosage of the pharmaceutical composition according to the present invention can vary depending on the formulation method, administration method, administration time and / or route of administration of the pharmaceutical composition, and can vary due to many factors, including the type and degree of reaction to be achieved by the administration of the pharmaceutical composition, the type of individual to be administered, age, weight, general health status, symptoms and severity of disease, sex, diet, excretion, drugs used simultaneously or at different times in the same individual, components of other compositions, and similar factors well known in the pharmaceutical field. Furthermore, a person with ordinary skill in the art can easily determine and prescribe a dosage that is effective for the intended treatment. For example, the daily dosage of the pharmaceutical composition according to the present invention is about 0.01 mg / kg to 1,000 mg / kg, preferably 0.1 mg / kg to 100 mg / kg, and can be administered once a day or divided into several doses per day.

[0043] The pharmaceutical composition according to the present invention may be administered once a day or in several divided doses per day. The pharmaceutical composition according to the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents. Taking all of the above factors into consideration, it is possible to administer an amount that provides the maximum effect with the minimum amount without side effects, and this can be easily determined by those skilled in the art.

[0044] The pharmaceutical compositions according to the present invention can be used in combination with various methods, such as hormone therapy and drug therapy, for the prevention or treatment of vaginitis and vulvitis.

[0045] As used in this invention, the term "administration" means introducing the pharmaceutical composition according to the present invention to a patient by some appropriate method, and the route and method of administration of the pharmaceutical composition according to the present invention may be independent of each other. Furthermore, any route and method of administration can be followed without special restrictions, as long as the pharmaceutical composition reaches the target site.

[0046] The aforementioned pharmaceutical composition can be administered orally or parenterally, and can be formulated into a variety of appropriate dosage forms for oral or parenteral administration.

[0047] Non-limiting examples of oral formulations using the pharmaceutical composition according to the present invention include oily suspensions, lozenges, tablets, water-soluble suspensions, prepared powders, granules, emulsions, hard capsules, soft capsules, syrups, or elixirs.

[0048] To formulate the pharmaceutical composition according to the present invention for oral administration, binders such as sorbitol, mannitol, starch, amylopectin, cellulose, lactose, saccharose, or gelatin; lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate, or polyethylene glycol wax; excipients such as dicalcium phosphate; disintegrants such as corn starch or sweet potato starch can be used, and fragrances, syrups, and sweeteners can also be used. Furthermore, in the case of capsules, in addition to the substances mentioned above, liquid carriers such as fatty oils can be additionally used.

[0049] Methods for parenteral administration of the pharmaceutical composition according to the present invention include intramuscular administration, transdermal administration, intravenous administration, intraperitoneal administration, or subcutaneous administration. Methods of applying the composition to the diseased area, spraying, or inhaling are also available, but are not limited to these.

[0050] Non-limiting examples of parenteral formulations using the pharmaceutical composition according to the present invention include injection solutions, suppositories, ointments, topical powders, oils, respiratory inhalation powders, spray aerosols, creams, and the like.

[0051] To formulate the pharmaceutical composition according to the present invention for parenteral administration, sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, topical preparations, etc., can be used. As the non-aqueous solvent and suspension, vegetable oils such as olive oil, injectable esters such as propylene glycol, polyethylene glycol, and ethyl oleate may be used.

[0052] When formulating the pharmaceutical composition according to the present invention into an injectable solution, the pharmaceutical composition according to the present invention can be prepared as a solution or suspension by mixing it with water together with a stabilizer or buffer, and this can be formulated for unit administration in ampoules or vials.

[0053] When formulating the pharmaceutical composition according to the present invention into an aerosol, it can be combined with additives such as propellants so that the dispersed concentrate or wet powder is dispersed.

[0054] As another way to achieve the above objective, the present invention provides a health functional food for improving vaginitis or vulvitis containing Lactobacillus salivarius HHuMin-U (KCCM13001P).

[0055] The aforementioned "health functional foods" refer to foods manufactured and processed using raw materials or ingredients that have beneficial functional properties for the human body, as defined in Act No. 6727 on Health Functional Foods. "Functionality" means ingesting a food for the purpose of regulating nutrients in relation to the structure and function of the human body, or obtaining beneficial effects for health purposes, such as physiological effects.

[0056] In the health functional food according to the present invention, the content of the bacterial strain according to the present invention can be varied depending on the condition of the recipient, the type of disease, the degree of progression, etc. If necessary, it may also be included in the total content of the food.

[0057] In the functional health food according to the present invention, the bacterial strain according to the present invention is preferably contained in an amount of 0.00001% to 50% by weight relative to the functional health food. If it is less than 0.00001% by weight, the effect will be negligible, and if it exceeds 50% by weight, the increase in effect will be negligible compared to the amount used, making it uneconomical.

[0058] The health functional foods according to the present invention may, for example, be probiotics, synbiotics, and postbiotic compositions. They may also be manufactured in any one form selected from noodles, gums, dairy products, ice cream, meats, grains, caffeine beverages, general beverages, chocolate, bread, snacks, confectionery, candy, pizza, jelly, sauces, alcoholic beverages, alcoholic drinks, vitamin complexes, and other health supplements, but are not necessarily limited to these.

[0059] When using the health functional food according to the present invention as a food additive, it can be added as is or used together with other foods or food components, and can be used appropriately by conventional methods.

[0060] The health functional food according to the present invention may typically contain additional ingredients that can improve odor, taste, appearance, etc. For example, it may contain biotin, folate, pantothenic acid, vitamins A, C, D, E, B1, B2, B6, B12, niacin, etc. It may also contain minerals such as chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), zinc (Zn), iron (Fe), and calcium (Ca). It may also contain amino acids such as cysteine, valine, lysine, and tryptophan. In addition, food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), colorants (tar dyes, etc.), color fixatives (sodium nitrite, etc.), bleaching agents (sodium sulfite), disinfectants (bleaching powder and high-grade bleaching powder, sodium hypochlorite, etc.), leavening agents (alum, potassium bitartrate, etc.), fortifiers, emulsifiers, thickeners, coating agents, antioxidants [butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), etc.], seasonings (MSG, monosodium glutamate, etc.), sweeteners (dulcin, cyclamic acid, saccharin, sodium, etc.), flavorings (vanillin, lactones, etc.), gum bases, antifoaming agents, solvents, and improvers may be added. The aforementioned additives may be selected and used in appropriate amounts depending on the type of food.

[0061] As yet another mode for achieving the above objective, the present invention provides a topical skin preparation for improving vaginitis or vulvitis, comprising Lactobacillus salivarius HHuMin-U (KCCM13001P).

[0062] In this invention, the term "improvement" means all actions in which vaginitis and vulvitis are improved or beneficially altered by the administration of the composition of this invention.

[0063] Examples of topical skin preparations according to the present invention include, but are not limited to, creams, gels, patches, sprays, ointments, plasters, lotions, liniments, pastes, and cataplasms.

[0064] Other ingredients that may be added include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, disinfectants, antioxidants, plant extracts, pH adjusters, alcohols, colorants, fragrances, blood circulation promoters, cooling agents, antiperspirants, and purified water. Examples of oils and fats include ester-based oils and fats, hydrocarbon-based oils and fats, silicone-based oils and fats, fluorinated oils and fats, animal fats and fats, and vegetable oils and fats.

[0065] Ester-based oils include glyceryl tri-2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, butyl stearate, ethyl linolenate, isopropyl linolenate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldodecyl myristate, isocetyl isostearate, diethyl sebacate, diisopropyl adipate, and neo Isoalkyl pentanoate, caprylic / capric triglyceride, trimethylolpropane tri-2-ethylhexanoate, trimethylolpropane triisostearate, pentaerythritol tetra-2-ethylhexanoate, cetyl caprylate, decyl laurate, hexyl laurate, decyl myristate, myristyl myristate, cetyl myristate, stearyl stearate, decyl oleate, cetyl ricinoleate, isostearyl laurate, isotridecyl myristate, isocetyl palmitate, octyl stearate, stearin Isocetyl acid, isodecyl oleate, octyldodecyl oleate, octyldodecyl linolenate, isopropyl isostearate, cetostearyl 2-ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprate, di(caprylic / capric acid)propylene glycol, propylene glycol dicaprylate, neopentyl glycol dicaprate, neopentyl glycol dioctanoate, glyceryl tricaprylate, tri Glyceryl undecylate, glyceryl triisopalmitate, glyceryl triisostearate, octyldodecyl neopentanoate, isostearyl octanoate, octyl isononanoate, hexyldecyl neodecanoate, octyldodecyl neodecanoate, isocetyl isostearate, isostearyl isostearate, octyldecyl isostearate, polyglycerin oleate, polyglycerin isostearate, triisocetyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate,Examples of ester compounds include cetyl lactate, octyldecyl lactate, triethyl citrate, acetyltriethyl citrate, acetyltributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di-2-ethylhexyl succinate, diisobutyl adipate, diisopropyl sebacate, dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, phytosteryl isostearate, phytosteryl oleate, isocetyl 12-stearoylhydroxystearate, stearyl 12-stearoylhydroxystearate, and isostearyl 12-stearoylhydroxystearate.

[0066] Examples of hydrocarbon-based oils and fats include squalene, liquid paraffin, α-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybutene, microcrystalline wax, and petrolatum.

[0067] Examples of silicone-based oils include polymethyl silicone, methylphenyl silicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane-methylcetyloxysiloxane copolymer, dimethylsiloxane-methylstearoxane copolymer, alkyl-modified silicone oil, and amino-modified silicone oil.

[0068] Examples of fluorinated oils and fats include perfluoropolyethers.

[0069] Examples of animal or vegetable oils include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, safflower oil, soybean oil, corn oil, rapeseed oil, apricot kernel oil, palm kernel oil, palm oil, castor oil, sunflower seed oil, grape seed oil, cottonseed oil, coconut oil, kukui nut oil, wheat germ oil, rice germ oil, shea butter, evening primrose oil, macadamia nut oil, meadowfoam oil, egg yolk oil, beef tallow, hemp oil, mink oil, orange roughy oil, jojoba oil, candelilla wax, carnauba wax, liquid lanolin, and hydrogenated castor oil.

[0070] Examples of humectants include water-soluble low-molecular-weight humectants, lipid-soluble molecular humectants, water-soluble polymers, and lipid-soluble polymers.

[0071] Examples of water-soluble low molecular weight humectants include serine, glutamine, sorbitol, mannitol, pyrrolidone sodium carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B (degree of polymerization n=2 or higher), polypropylene glycol (degree of polymerization n=2 or higher), polyglycerin B (degree of polymerization n=2 or higher), lactic acid, and lactate salts.

[0072] Examples of lipid-soluble low-molecular-weight humectants include cholesterol and cholesterol esters.

[0073] Examples of water-soluble polymers include carboxyvinyl polymer, polyaspartate, tragacanth, xanthan gum, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, water-soluble chitin, chitosan, and dextrin.

[0074] Examples of lipid-soluble polymers include polyvinylpyrrolidone-eicosene copolymers, polyvinylpyrrolidone-hexadecene copolymers, nitrocellulose, dextrin fatty acid esters, and high-molecular-weight silicones.

[0075] Examples of emollients include long-chain acyl glutamate cholesteryl esters, cholesteryl hydroxystearate, 12-hydroxystearic acid, stearic acid, rosinic acid, and lanolin fatty acid cholesteryl esters.

[0076] Examples of surfactants include nonionic surfactants, anionic surfactants, cationic surfactants, and positive surfactants.

[0077] Examples of nonionic surfactants include self-emulsifying glyceryl monostearate, propylene glycol fatty acid esters, glycerin fatty acid esters, polyglycerin fatty acid esters, sorbitan fatty acid esters, POE (polyoxyethylene) sorbitan fatty acid esters, POE sorbitan fatty acid esters, POE glycerin fatty acid esters, POE alkyl ethers, POE fatty acid esters, POE hydrogenated castor oil, POE castor oil, POE-POP (polyoxyethylene-polyoxypropylene) copolymers, POE-POP alkyl ethers, polyether-modified silicones, lauric acid alkanolamides, alkylamine oxides, and hydrogenated soybean phospholipids.

[0078] Examples of anionic surfactants include fatty acid soaps, α-acyl sulfonates, alkyl sulfonates, alkylallyl sulfonates, alkylnaphthalene sulfonates, alkyl sulfates, POE alkyl ether sulfates, alkylamide sulfates, alkyl phosphates, POE alkyl phosphates, alkylamide phosphates, alkylylalkyl taurates, N-acyl amino acid salts, POE alkyl ether carboxylates, alkyl sulfosuccinates, alkyl sulfoacetate sodium, acylated hydrolyzed collagen peptides, and perfluoroalkyl phosphate esters.

[0079] Examples of cationic surfactants include alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, benzalkonium chloride, diethylaminoethylamide stearate, dimethylaminopropylamide stearate, and quaternary ammonium salts of lanolin derivatives.

[0080] Examples of positively positive surfactants include carboxybetaine type, amidebetaine type, sulfobetaine type, hydroxysulfobetaine type, amidesulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, and amideamine type.

[0081] Examples of organic and inorganic pigments include inorganic pigments such as silicic acid, anhydrous silicic acid, magnesium silicate, talc, sericite, mica, kaolin, red iron oxide, clay, bentonite, titanium-coated mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, aluminum oxide, calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine, chromium oxide, chromium hydroxide, calamine, and composites thereof; organic pigments such as polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenolic resin, fluororesin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene-styrene copolymer, silk powder, cellulose, CI pigment yellow, CI pigment orange, and composite pigments of these inorganic and organic pigments.

[0082] Examples of organic powders include metal soaps such as calcium stearate; alkyl phosphate metal salts such as sodium zinc cetyl phosphate, zinc lauryl phosphate, and calcium lauryl phosphate; acyl amino acid polyvalent metal salts such as N-lauroyl-β-alanine calcium, N-lauroyl-β-alanine zinc, and N-lauroylglycine calcium; amide sulfonic acid polyvalent metal salts such as N-lauroyl-taurine calcium and N-palmitoyl-taurine calcium; N-acyl basic amino acids such as N-ε-lauroyl-L-lysine, N-ε-palmitoyl lysine, N-α-palmitoylornithine, N-α-lauroylarginine, and N-α-hydrogenated beef tallow fatty acid acylarginine; N-acyl polypeptides such as N-lauroyl glycylglycine; α-amino fatty acids such as α-aminocaprylic acid and α-aminolauric acid; and polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, divinylbenzene-styrene copolymer, and tetrafluoroethylene.

[0083] UV absorbers include para-aminobenzoic acid, ethyl para-aminobenzoate, amyl para-aminobenzoate, octyl para-aminobenzoate, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butylphenyl salicylate, homomentyl salicylate, benzyl cinnamate, 2-ethoxyethyl para-methoxycinnamate, octyl para-methoxycinnamate, mono-2-ethylhexaneglyceryl dipara-methoxycinnamate, isopropyl para-methoxycinnamate, diisopropyl / diisopropyl cinnamic acid ester mixture, and urocanin. Examples include acids, ethyl urocanate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenone sulfonic acid and its salts, dihydroxymethoxybenzophenone, sodium dihydroxymethoxybenzophenone disulfonate, dihydroxybenzophenone, tetrahydroxybenzophenone, 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-trianilino-p-(carbo-2'-ethylhexyl-1'-oxy)-1,3,5-triazine, and 2-(2-hydroxy-5-methylphenyl)benzotriazole.

[0084] Examples of disinfectants include hinokitiol, triclosan, trichlorohydroxydiphenyl ether, chlorhexidine gluconate, phenoxyethanol, resorcinol, isopropylmethylphenol, azulene, salicylic acid, zinc pyrithione, benzalkonium chloride, photosensitizer 301, mononitroguaiacol sodium, and undecylenic acid.

[0085] Examples of antioxidants include butylhydroxyanisole, propyl gallate, and erythorbic acid.

[0086] Examples of pH adjusters include citric acid, sodium citrate, malic acid, sodium malate, fumaric acid, sodium fumarate, succinic acid, sodium succinate, sodium hydroxide, and sodium monohydrogen phosphate.

[0087] Examples of alcohols include higher alcohols such as cetyl alcohol.

[0088] Furthermore, other ingredients that can be added are not limited to those mentioned above, and any of the aforementioned ingredients can be added within a range that does not impair the purpose and effects of the present invention.

[0089] The topical skin preparation according to the present invention can vary considerably depending on many factors, including the activity of the specific active ingredient used, age, weight, general health, sex, diet, application time, drug formulation, and the severity of the specific disease being prevented or treated. Furthermore, the dosage of the topical skin preparation may vary depending on the patient's condition, weight, disease severity, drug form, route of application, and duration, but can be appropriately selected by those skilled in the art. Preferably, taking all of the above factors into consideration, the amount that provides the maximum effect with the minimum amount without side effects can be applied, preferably an effective dose of 1 to 10,000 μg / kg body weight / day, more preferably 10 to 1,000 mg / kg body weight / day, which can be administered repeatedly several times a day. The aforementioned dosage does not limit the scope of the present invention in any respect.

[0090] As another way to achieve the above objective, the present invention provides a cleansing composition for improving vaginitis or vulvitis, comprising Lactobacillus salivarius HHuMin-U (KCCM13001P).

[0091] In the present invention, the bacterial strain is preferably contained in an amount of 0.00001% to 30.0% by weight relative to the total weight of the cleansing agent composition, and more preferably in an amount of 0.01% to 10% by weight relative to the total weight of the cleansing agent composition. If the content of the bacterial strain is less than 0.00001% by weight, no improvement in vaginitis and vulvitis is observed, and if the content of the bacterial strain exceeds 30.0% by weight, no significant increase in effect due to increased content is observed.

[0092] On the other hand, the components included in the detergent composition of the present invention may include, as active ingredients, various components commonly used in detergent compositions, in addition to the bacterial strain of the present invention, such as antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances, and other common auxiliary agents and carriers.

[0093] The detergent composition of the present invention can be manufactured in any dosage form commonly produced in the industry, and can be, but is not limited to, solutions, suspensions, emulsions, pastes, gels, creams, soaps, surfactant-containing cleansers, oils, and sprays.

[0094] When the dosage form of the detergent composition of the present invention is a paste, cream, or gel, animal oils, vegetable oils, waxes, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide may be used as the carrier component.

[0095] When the dosage form of the detergent composition of the present invention is a solution or emulsion, a solvent, solubilizer, or emulsifier can be used as the carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, or fatty acid ester of sorbitan.

[0096] When the dosage form of the detergent composition of the present invention is a suspension, the carrier component may be a liquid diluent such as water, ethanol, or propylene glycol; a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, or polyoxyethylene sorbitan ester; or microcrystalline cellulose, aluminum methhydroxyl, bentonite, aga, or tracant.

[0097] When the dosage form of the detergent composition of the present invention is a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder can be used as the carrier component, and in particular when it is a spray, propellants such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether may be additionally included.

[0098] When the dosage form of the detergent composition of the present invention is a surfactant-containing cleanser, the carrier component may be an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinate monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamide betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

[0099] When the cleansing composition of the present invention is a soap, a surfactant-containing cleansing formulation, or a surfactant-free cleansing formulation, it can be applied to the skin and then wiped off, peeled off, or washed with water. Specific examples include, but are not limited to, liquid soap, powder soap, solid soap, and oil soap, and the surfactant-containing / non-containing cleansing formulations are cleansing cream, cleansing water, and cleansing gel.

[0100] The contents of the present invention will be described in more detail below through the following examples or experimental examples. However, the scope of the present invention is not limited to the following examples or experimental examples, but also includes equivalent variations of the technical idea thereto. [Examples]

[0101] [Example 1: Evaluation of the antimicrobial activity of Lactobacillus salivarius HHuMin-U strain against Candida albicans]

[0102] Candida albicans strain (KCTC2720), a representative strain that induces vaginitis and vulvitis, was cultured in YM medium under anaerobic conditions (85% N2, 10% H2, and 5% CO2).

[0103] Candidate strains, including Lactobacillus salivarius HHuMin-U strain (hereinafter referred to as "HMU"), were cultured in a jar fermenter (Fermentec, Korea) using modified MRS medium at 37°C, pH 5.5, and 150 rpm for 15 hours under anaerobic conditions (85% N2, 10% H2, and 5% CO2). The cultures were then centrifuged at 7,000 rpm for 15 minutes, and the supernatant was separated. The separated supernatant was filtered using a 0.20 μm filter.

[0104] The supernatant was added to Candida albicans culture at a concentration of 3-5% (v / v), and the degree of inhibition was confirmed by observing the growth curve. As a control group, culture media without inoculation of Lactobacillus salivarius HHuMin-U strain was used.

[0105] The experimental results confirmed that Lactobacillus salivarius HHuMin-U strain (HMU) exhibited superior antibacterial activity against Candida albicans using numerous strains (Figure 1). Figure 1 shows the results of the antibacterial activity of Lactobacillus salivarius HHuMin-U strain according to the present invention against Candida albicans strains. "CH88" in Figure 1 refers to the Lactobacillus paracasei (L. paracasei) CH88 strain, and the "numbers" represent unidentified candidate strains.

[0106] [Example 2: Evaluation of therapeutic efficacy of human vaginal epithelial cell lines for vaginitis and vulvitis]

[0107] In all experiments described below, Lactobacillus salivarius HHuMin-U strain was cultured in a jar fermenter (Fermentec, Korea) using modified MRS medium at 37°C, pH 5.5, and 150 rpm for 15 hours under anaerobic conditions (85% N2, 10% H2, and 5% CO2). The culture was then centrifuged at 7,000 rpm for 15 minutes, the supernatant was removed, and the resulting pellet was freeze-dried with a cryoprotectant before being ground into a pulverized powder, which was used as the sample.

[0108] 1. Evaluation of the adhesion inhibitory ability of Candida albicans strains to human vaginal epithelial cells (Adhesion assay)

[0109] Human vaginal epithelial cell lines were dispensed into 6-well cell culture plates and cultured until a monolayer was formed, then heated in HMU 5×10⁶. 7 , 1 x 10 8After treating with CFU or PBS for 4 hours, Candida albicans strain 4 × 10 4 I received CFU (Chronic Fever-Related Mutual Aid) vaccination.

[0110] After culturing for 3 hours, the culture medium was removed, and any non-attached Candida albicans strains were removed by PBS washing. Vaginal epithelial cells were removed by adding 1 ml of distilled water to the washed wells and allowing them to react for 15 minutes. After continuous dilution while collecting all the solutions, the mixture was streaked onto Sabouraud dextrose agar plates, and the CFU of Candida albicans strains was measured.

[0111] The experimental results showed that treatment with the Lactobacillus salivarius HHuMin-U strain reduced the adhesion of Candida albicans strains to vaginal epithelial cells (Figure 2). Figure 2 shows the results of evaluating the effect of the Lactobacillus salivarius HHuMin-U strain according to the present invention on the adhesion of Candida albicans strains to vaginal epithelial cells.

[0112] 2. Evaluation of the ability of Candida albicans strains to kill human vaginal epithelial cells (LDH assay)

[0113] After dispensing human vaginal epithelial cell lines into 6-well cell culture plates, they were cultured until they became a monolayer, and then subjected to HMU 5×10⁶. 7 , 1 x 10 8 After treating with CFU or PBS for 4 hours, 2 x 10⁻¹⁰ Candida albicans strains 6 CFU was inoculated and cultured.

[0114] Subsequently, the culture medium was collected at 48 and 72 hours, and the LDH level of the medium was measured using the Cyquant LDH Cytotoxicity Assay Kit (Thermo Fisher Scientific) to confirm the relative degree of vaginal epithelial cell death.

[0115] The experimental results showed that treatment with the Lactobacillus salivarius HHuMin-U strain reduced the death of vaginal epithelial cells caused by Candida albicans strains (Figure 3). Figure 3 shows the results of evaluating the effect of the Lactobacillus salivarius HHuMin-U strain according to the present invention on the ability of Candida albicans strains to suppress the death of vaginal epithelial cells.

[0116] 3. Evaluation of the antibacterial activity of Candida albicans strains in human vaginal epithelial cells (Growth inhibition assay)

[0117] After dispensing human vaginal epithelial cell lines into 6-well cell culture plates, they were cultured until they became a monolayer, and then subjected to HMU 1×10⁶. 8 After treating with CFU or PBS for 6 hours, the culture medium was collected.

[0118] Subsequently, the collected culture medium was centrifuged at 3,000 rpm, and the supernatant was collected for use in the experiment. 150 μl of this supernatant was added to each 96-well plate, and then 4 × 10⁶ Candida albicans strains were added. 4 CFU was inoculated. This was cultured at 37°C, and the OD value was measured at 0, 12, and 24 hours to determine the degree of growth of the Candida albicans strain.

[0119] The experimental results confirmed that treatment with the Lactobacillus salivarius HHuMin-U strain suppressed the growth of Candida albicans strains in human vaginal epithelial cells (Figure 4). Figure 4 shows the results of evaluating the effect of the Lactobacillus salivarius HHuMin-U strain according to the present invention on the growth of Candida albicans strains in vaginal epithelial cells.

[0120] 4. Expression analysis of antimicrobial peptides and immunoactive substances

[0121] I. Antimicrobial peptide expression analysis (RT-qPCR)

[0122] After dispensing a human vaginal epithelial cell line into a 6-well cell culture plate, it was cultured until it became a monolayer, and HMU 5×10 7 、1×10 8 CFU or PBS was treated for 6 hours. Then, the medium was removed, washed twice with cold PBS, and mRNA was isolated. cDNA was synthesized from the isolated mRNA, and RT-qPCR was performed using this.

[0123] As a result of the experiment, it was confirmed that the treatment with Lactobacillus salivarius HHuMin-U strain increased the expression of antimicrobial peptides in human vaginal epithelial cells (Figure 5). Figure 5 shows the results of evaluating the effect of the Lactobacillus salivarius HHuMin-U strain according to the present invention on the expression of antimicrobial peptides in vaginal epithelial cells (upper panel: mRNA, lower panel: protein).

[0124] II. Analysis of the expression of immunologically active substances (ELISA)

[0125] After dispensing a human vaginal epithelial cell line into a 6-well cell culture plate, it was cultured until it became a monolayer, and HMU 5×10 7 、1×10 8 CFU or PBS was treated for 6 hours. Then, centrifugation was performed at 13,000 rpm while recovering the medium, and the obtained supernatant was used for ELISA analysis.

[0126] As a result of the experiment, it was confirmed that the treatment with Lactobacillus salivarius HHuMin-U strain increased the expression of immunologically active substances in human vaginal epithelial cells (Figure  6). Figure 6 shows the results of evaluating the effect of the Lactobacillus salivarius HHuMin-U strain according to the present invention on the expression of immunologically active substances in vaginal epithelial cells (upper panel: mRNA, lower panel: protein).

[0127] [Example 3: Evaluation of the therapeutic efficacy of vaginitis and vulvitis in an animal model]

[0128] In all experiments described below, Lactobacillus salivarius HHuMin-U strain (hereinafter referred to as "HMU") was cultured in a jar fermenter (Fermentec, Korea) using modified MRS medium at 37°C, pH 5.5, and 150 rpm for 15 hours under anaerobic conditions (85% N2, 10% H2, and 5% CO2). The culture was then centrifuged at 7,000 rpm for 15 minutes, the supernatant was removed, and the pellet was freeze-dried with a cryoprotectant before being ground into a pulverized powder, which was used as the sample.

[0129] 1. Oral administration

[0130] Six-week-old female C57BL / 6 mice were acclimatized for one week before the experiment. Each group was treated with either PBS or HMU (1 × 10⁶). 7 , 1 x 10 8 CFU was administered orally at a dose of 100 μL each day for a total of 13 days. On day 8 of oral administration, 0.5 mg / 100 μL of β-estradiol dissolved in sesame oil was administered subcutaneously, and on day 11, 5 × 10⁻¹⁰ CFU was administered. 5 Infection was induced by inoculating 20 μL of PBS into the vagina.

[0131] The experiment was completed on day 14, three days after infection. The vagina was washed with 100 μL of PBS, and the washing solution was successively diluted and streaked onto Sabouraud dextrose agar. The CFU of Candida albicans strains was measured, and vaginal tissue was collected and analyzed. For the analysis of vaginal tissue, the tissue was fixed with 10% paraformaldehyde, a paraffin block was prepared, which was then made into a tissue slide and stained with periodic acid Schiff staining (PAS staining). This staining allows for the identification of Candida albicans strains that have infiltrated the tissue.

[0132] The experimental results showed a significant decrease in the CFU of Candida albicans strains in the vaginal douche, and infection was confirmed to be suppressed by tissue PAS staining (Figure 7). Figure 7 shows the results of evaluating the effect of oral administration of Lactobacillus salivarius HHuMin-U strain according to the present invention on the improvement and treatment of vaginitis (A: change in body weight, B: measurement of C. albicans CFU in vaginal douche, C: PAS staining of vaginal tissue).

[0133] 2. Vaginal administration

[0134] Six-week-old female C57BL / 6 mice were acclimatized for one week before the experiment. Each group was treated with either PBS or HMU (1 × 10⁶). 6 CFU was administered vaginally for 5 days. On the 3rd day of vaginal administration, 0.5 mg / 100 μL of β-estradiol dissolved in sesame oil was subcutaneously injected, and on the 6th day, 5 × 10⁻¹⁵ Candida albicans strains were administered. 5 Infection was induced by inoculating 20 μL of PBS into the vagina.

[0135] The experiment was concluded on the third day of infection. The vagina was washed with 100 μL of PBS, and the washing solution was successively diluted and then streaked onto Sabouraud dextrose agar to measure the CFU of Candida albicans strains.

[0136] The experimental results confirmed a significant decrease in the CFU of Candida albicans strains in the vaginal douche (Figure 8). Figure 8 shows the results of evaluating the effect of vaginal administration of Lactobacillus salivarius HHuMin-U strain according to the present invention on the improvement and treatment of vaginitis.

Claims

1. A pharmaceutical composition for the prevention or treatment of vaginitis or vulvitis, comprising Lactobacillus salivarius HHuMin-U (KCCM13001P) as the active ingredient.

2. The pharmaceutical composition according to claim 1, wherein the bacterial strain is one or more selected from live Lactobacillus salivarius HHuMin-U (KCCM13001P) strain, its crushed product, its culture medium, its culture, its extract, and its dead cells.

3. The pharmaceutical composition according to claim 1, wherein the vaginitis or vulvitis is one selected from bacterial vaginosis, trichomoniasis, candidiasis, atrophic vaginitis, herpes simplex virus infection, and vulvar warts.

4. A health supplement containing Lactobacillus salivarius HHuMin-U (KCCM13001P) for the improvement of vaginitis or vulvitis.

5. A topical skin preparation containing Lactobacillus salivarius HHuMin-U (KCCM13001P) for the improvement of vaginitis or vulvitis.

6. A cleansing composition for improving vaginitis or vulvitis, containing Lactobacillus salivarius HHuMin-U (KCCM13001P).

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