WO2022145960A1 - Souche de weissella cibaria et son utilisation - Google Patents

Souche de weissella cibaria et son utilisation Download PDF

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WO2022145960A1
WO2022145960A1 PCT/KR2021/020031 KR2021020031W WO2022145960A1 WO 2022145960 A1 WO2022145960 A1 WO 2022145960A1 KR 2021020031 W KR2021020031 W KR 2021020031W WO 2022145960 A1 WO2022145960 A1 WO 2022145960A1
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Prior art keywords
strain
skin
composition
oral
culture medium
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PCT/KR2021/020031
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English (en)
Korean (ko)
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WO2022145960A9 (fr
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조형우
백채윤
이동걸
김미선
김민지
강승현
박명삼
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코스맥스 주식회사
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Publication of WO2022145960A1 publication Critical patent/WO2022145960A1/fr
Publication of WO2022145960A9 publication Critical patent/WO2022145960A9/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q11/00—Preparations for care of the teeth, of the oral cavity or of dentures; Dentifrices, e.g. toothpastes; Mouth rinses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales

Definitions

  • the skin ecosystem provides various types of habitat for microorganisms, and a wide range of microorganisms live there. They form a symbiotic relationship with the human host, and are known to have many positive effects on the host.
  • the skin constitutes various types of habitats, such as depressions and specialized crevices, and helps a wide range of microorganisms to grow. Basically, the skin forms a physical barrier and helps to defend against potential hazards and toxic substances from the outside.
  • the skin becomes a connection point with the external environment and is also a gathering place for various microorganisms (fungi, bacteria, viruses and small larvae). In accordance with the selection of physical and chemical functions, microorganisms adapt to specialized niches to prepare habitats.
  • the skin is cold, acidic, and remains dry. Structurally, the epidermis forms a skin barrier, blocks the penetration of microorganisms and toxins, and plays an important role in maintaining moisture.
  • the uppermost layer of the epidermis is composed of the stratum corneum.
  • the epidermis has a so-called 'brick and mortar structure', and the skin tissue undergoes a continuous self-healing process, and the squames that have undergone the final differentiation process repeat the process of constantly being removed from the skin tissue.
  • the lactic acid bacteria are used in the manufacturing process of fermented foods such as kimchi or yogurt, and the lactic acid bacteria used in food include Enterococcus genus, Lactobacillus genus, Lactococcus genus, Leukonostok ( Leuconostoc ) genus, Streptococcus genus, Weissella genus, and the like.
  • these lactic acid bacteria have a problem in that when the lactic acid bacteria are administered to the oral cavity, they are swallowed directly into the esophagus while being diluted with saliva like metal salts or disinfectants, making it difficult to remain in the oral cavity.
  • Lactobacillus acidophilus Lactobacillus acidophilus
  • Lactobacillus casei Lactobacillus casei
  • Lactobacillus salivarius Lactobacillus salivarius
  • strong acids cause dental caries, there is a problem in that oral hygiene is not good if these bacteria are administered to the oral cavity for a long time. Therefore, there is a need for the development of microorganisms for reducing bad breath.
  • One aspect is to provide a CXO-1 strain belonging to the Weissella sp .
  • Another aspect is to provide a lysate or culture solution of the strain.
  • Another aspect is to provide a cosmetic composition
  • a cosmetic composition comprising the strain, a lysate thereof, a culture solution, an extract of the culture solution, or a mixture thereof.
  • Another aspect is to provide a pharmaceutical composition for the treatment of wounds comprising the strain, a lysate thereof, a culture solution, an extract of the culture solution, or a mixture thereof.
  • Another aspect is to provide an oral composition
  • an oral composition comprising the strain, a lysate thereof, a culture solution, an extract of the culture solution, or a mixture thereof.
  • Another aspect is to provide a pharmaceutical composition for the prevention or treatment of oral diseases comprising the above strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof.
  • Another aspect is to provide a method of treating a wound comprising administering to a subject a composition comprising the strain, a lysate thereof, a culture solution, an extract of the culture solution, or a mixture thereof.
  • Another aspect is to provide a method of treating oral diseases comprising administering to an individual a composition comprising the strain, a lysate thereof, a culture solution, an extract of the culture solution, or a mixture thereof.
  • Another aspect is to provide a wound treatment, prevention or treatment of oral diseases of the composition comprising the strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof.
  • the Weissella cibaria strain may be a strain deposited with accession number KCCM12889P.
  • the Weissellacibaria strain may be a strain comprising 16s rRNA of SEQ ID NO: 1.
  • the strain increases the expression of anti-aging and anti-wrinkle factors, for example, COL1A1 or decreases the expression of MMP-1, and hyaluronan synthase 3 (Hyaluronan synthase), which is a factor related to skin moisturizing and skin barrier 3, HSA3) was confirmed to increase the expression of or filaggrin.
  • HSA3 hyaluronan synthase 3
  • the strain according to an aspect may have a skin beauty improvement effect, for example, skin aging inhibition, skin barrier strengthening, skin wrinkle inhibition, or skin moisturizing effect.
  • Another aspect provides a lysate of the strain, a culture solution, or an extract of the culture solution. Specific details of the strain are the same as described above.
  • culture medium may be used interchangeably with “culture supernatant”, “conditioned culture medium” or “conditioned medium”, and may be used to provide nutrients so that the B. sibira strain can grow and survive in vitro. It may mean the entire medium including the strain obtained by culturing the strain for a certain period of time, its metabolites, and extra nutrients in a medium that can.
  • the culture solution may refer to a culture solution obtained by culturing the strain in which the cells are removed from the culture solution.
  • the liquid from which the cells have been removed among the above culture solution is also called “supernatant”, and the culture solution is left for a certain period of time to take only the liquid from the upper layer except for the part that has sunk to the lower layer, remove the cells through filtration, or centrifuge the culture solution to the lower part It can be obtained by removing the precipitation of and taking only the liquid at the top.
  • the "cell” of the present invention refers to the strain itself of the present invention, and includes the strain isolated and selected from a skin sample, or the strain isolated from the culture solution by culturing the strain. The cells can be obtained by centrifuging the culture medium to take the part that has sunk to the lower layer, or by gravity sink to the lower layer of the culture solution, leaving it still for a certain period of time and then removing the upper liquid It can be obtained.
  • the culture medium may include the culture medium itself obtained by culturing the strain, its concentrate, or a freeze-dried product or a culture supernatant obtained by removing the strain from the culture medium, its concentrate or freeze-dried product.
  • the culture medium is obtained by culturing the Weissella sibaira strain in a medium (eg, R2A medium or TSA medium) at any temperature of more than 10° C. or less than 40° C. for a certain period of time, for example, 4 to 50 hours.
  • a medium eg, R2A medium or TSA medium
  • the culture supernatant of the strain can be obtained by centrifuging or filtering the strain culture solution to remove the strain.
  • the concentrate may be obtained by concentrating the supernatant obtained after filtering the strain culture solution itself, or the culture solution using a centrifuge or filter.
  • the culture medium and culture conditions for culturing the Weissella sibaira strain may be appropriately selected or modified by those of ordinary skill in the art.
  • lysate may refer to a product obtained by disrupting the cell wall of the strain itself by chemical or physical force.
  • culture solution extract means extraction from the culture solution or its concentrate, and includes an extract, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or these prepared or purified products, and fractions obtained by fractionation thereof. can do.
  • Another aspect provides the use of the strain, a lysate of the strain, a culture solution, or an extract of the culture solution. Specifically, it provides a composition comprising a Weissella sibaira strain, a lysate thereof, a culture solution or an extract of the culture solution thereof.
  • Uses of the strain may include improving skin condition, improving skin beauty, preventing, improving or treating skin diseases.
  • the skin condition improvement or skin beauty improvement may be skin aging inhibition or improvement, anti-wrinkle, skin elasticity, skin regeneration, skin barrier strengthening, skin moisturizing or wound improvement.
  • skin aging refers to the tangible and intangible changes that appear on the skin with age, for example, a phenomenon in which the thickness of the epidermis becomes thin, the number of cells or blood vessels in the dermis, DNA damage repair ability, cell replacement cycle , wound recovery, skin barrier function, epidermal moisture retention, sweat secretion, sebum secretion, vitamin D production, physical damage defense, chemical removal ability, immune response, sensory function, and decreased body temperature control.
  • the strain or its culture medium may be for improving skin aging caused by extrinsic factors or intrinsic factors.
  • the extrinsic factor refers to various external factors, such as ultraviolet light (light), and the intrinsic factor is also referred to as a chronological factor and refers to a factor mainly caused by the passage of time. That is, the skin aging specifically refers to not only the premature aging symptoms induced by external stimuli such as ultraviolet rays, pollution, cigarette smoke, chemical substances, etc., but also a natural aging phenomenon that occurs as the proliferation of skin cells decreases with aging. It is a concept that includes all of wrinkles, elasticity reduction, skin sagging and dryness. In addition, wrinkles include those that cause wrinkles by changing the components constituting the skin tissue by stimulation by changes in internal and external factors.
  • the aging may be photoaging.
  • photoaging is a phenomenon induced by external environmental factors, and the most representative factor is ultraviolet rays. Ultraviolet rays cause damage to biological components such as activation of proteolytic enzymes, chain cleavage of matrix proteins, and abnormal cross-linking, and repetition of these mechanisms leads to apparent skin aging.
  • wrinkle refers to a state in which the elasticity of the skin is lost and loosened, and for example, the skin may be folded.
  • skin wrinkle prevention or improvement may refer to any action of preventing or improving wrinkles by inhibiting the expression of factors related to wrinkles, or increasing the total amount of collagen.
  • skin barrier strengthening may refer to any action that enhances the function of the skin barrier that is located at the outermost part of the skin and prevents loss of moisture and nutrients.
  • damage to the skin barrier function may refer to any change that appears in the skin due to the deterioration or damage of the function of the skin barrier. For example, it may include increased skin wrinkles, dryness, dermatitis, atopic dermatitis, allergic dermatitis, acne, and the like.
  • skin moisturizing may refer to any action of maintaining skin moisture or preventing moisture loss.
  • wound is also called “wound” and means damage to the living body.
  • the loss may be due to external factors such as, for example, physical damage, radiation, stimulation by chemical substances, etc., proliferation of microorganisms, or internal tolerance such as stress.
  • the wounds may include all kinds of wounds, such as cleavage, puncture, cleavage, acne, fissure, bronchial, and school spear.
  • Wound improvement may refer to any action that lightens or reduces the extent of a wound on the face or body
  • wound healing may refer to a series of biological reactions to repair the wound.
  • the strain may be used together with other strains belonging to the genus Weissella having a skin improvement effect to show a synergistic effect.
  • the strain belonging to the genus Weissella is, for example, Weissella confuse ( Weissella confuse ), Weissella ubarum ) and the like.
  • the composition comprises 0.001 wt% to 80 wt%, for example, 0.01 wt% to 60 wt%, 0.01 wt% to 40 wt%, 0.01 wt% to 30 wt%, 0.01 wt% to 20 wt%, based on the total weight of the composition %, 0.01% to 10%, 0.01% to 5%, 0.05% to 60%, 0.05% to 40%, 0.05% to 30%, 0.05% to 20% by weight, 0.05% to 10% by weight, 0.05% to 5% by weight, 0.1% to 60% by weight, 0.1% to 40% by weight, 0.1% to 30% by weight, 0.1% to 20% by weight, 0.1% by weight % to 10% by weight, or 0.1% to 5% by weight of the strain, a lysate thereof, a culture medium, or an extract of the culture medium thereof.
  • the composition may be a cosmetic composition.
  • the cosmetic composition may have a cosmetic formulation of, for example, a softening lotion, a nourishing lotion, a massage cream, a nourishing cream, an essence, a pack, a gel, an ampoule, or a skin adhesion type.
  • Components included in the cosmetic composition may include components commonly used in cosmetic compositions in addition to the composition as an active ingredient, for example, conventional adjuvants and carriers such as stabilizers, solubilizers, vitamins, pigments and fragrances. may include
  • the composition may be a composition for external application to the skin.
  • the external preparation for skin may be a cream, gel, ointment, skin emulsifier, skin suspension, transdermally delivered patch, drug-containing bandage, lotion, or a combination thereof.
  • the external preparation for skin is a component usually used in external preparations for skin such as cosmetics or pharmaceuticals, for example, an aqueous component, an oily component, a powder component, alcohol, a moisturizer, a thickener, an ultraviolet absorber, a whitening agent, a preservative, an antioxidant, a surfactant, a fragrance , colorant, various skin nutrients, or a combination thereof may be appropriately formulated as needed.
  • the external preparation for skin includes metal-blocking agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid, caffeine, tannin, belapamil, licorice extract, glablidine, and kaline.
  • metal-blocking agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid, caffeine, tannin, belapamil, licorice extract, glablidine, and kaline.
  • Fruit hot water extracts, various herbal medicines, tocopherol acetate, glitylittic acid, tranexamic acid and derivatives or salts thereof, vitamin C, magnesium ascorbate phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, Sugars, such as trehalose, etc. can be mix
  • the composition may be a pharmaceutical composition.
  • the pharmaceutical composition may further include a pharmaceutically acceptable diluent or carrier.
  • the diluent may be lactose, corn starch, soybean oil, microcrystalline cellulose, or mannitol, and the lubricant may be magnesium stearate, talc, or a combination thereof.
  • the carrier may be an excipient, a disintegrant, a binder, a lubricant, or a combination thereof.
  • the excipient may be microcrystalline cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof.
  • the disintegrant may be carboxymethyl cellulose calcium, sodium starch glycolate, anhydrous calcium monohydrogen phosphate, or a combination thereof.
  • the binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof.
  • the lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.
  • the pharmaceutical composition may be formulated as an oral or parenteral dosage form.
  • Oral dosage forms may be granules, powders, solutions, tablets, capsules, dry syrups, or a combination thereof.
  • the parenteral dosage form may be an injection.
  • the composition may be a health functional food composition.
  • the health functional food composition may be used alone or in combination with other foods or food ingredients of the strain or its culture, and may be appropriately used according to a conventional method.
  • the mixing amount of the active ingredient may be appropriately determined depending on the purpose of use (prophylactic, health or therapeutic treatment).
  • the composition of the present specification may be added in an amount of 15 parts by weight or less based on the raw material.
  • the beverage composition may contain various flavoring agents or natural carbohydrates as an additional component like a conventional beverage.
  • the natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol.
  • sweetener natural sweeteners such as taumartin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like can be used.
  • the health food composition may also be added to nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonated beverages carbonation agent used, or a combination thereof.
  • the health functional food composition may also contain natural fruit juice, fruit juice beverage, fruit flesh for the production of vegetable beverage, or a combination thereof.
  • Another aspect provides a method of preventing, ameliorating or treating a skin condition in a subject comprising treating or administering to the subject in need thereof an effective amount of the composition.
  • Specific details of the skin condition are the same as described above.
  • Another aspect provides a pharmaceutical composition for wound treatment comprising the strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof as an active ingredient.
  • Another aspect provides an external composition for wound treatment comprising the strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof as an active ingredient.
  • Another aspect provides a method for treating a wound comprising administering to a subject in need thereof an effective amount of the strain, a lysate thereof, a culture solution, an extract of the culture solution, or a mixture thereof. The specific details of the wound are the same as described above.
  • administering As used herein, the terms “administering”, “introducing”, and “implanting” are used interchangeably and into a subject by a method or route that results in at least partial localization of a composition according to one embodiment to a desired site. It may mean the arrangement of the composition according to one embodiment of the.
  • Administration may be administered by methods known in the art. Administration may be administered directly to a subject by any means, for example, intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal or subcutaneous administration. can The administration may be systemically or locally.
  • the subject may be a mammal, such as a human, cow, horse, pig, dog, sheep, goat, or cat.
  • the subject may be an individual in need of skin beauty improvement, for example, skin moisturizing, skin barrier strengthening, skin wrinkle improvement effect.
  • the administration is 0.1 mg to 1,000 mg, for example, 0.1 mg to 500 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 1 mg to 1,000 mg, 1 mg to 500 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 5 mg to 1,000 mg, 5 mg to 500 mg, 5 mg to 100 mg, 5 mg to 50 mg , 5 mg to 25 mg, 10 mg to 1,000 mg, 10 mg to 500 mg, 10 mg to 100 mg, 10 mg to 50 mg, or 10 mg to 25 mg may be administered.
  • the dosage may be prescribed in various ways depending on factors such as formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate and reaction sensitivity, and those skilled in the art
  • the dosage may be appropriately adjusted in consideration of these factors.
  • the number of administration may be once a day or twice or more within the range of clinically acceptable side effects, and may be administered to one or two or more places for the site of administration, and total daily or at intervals of 2 to 5 days
  • the number of days of administration may range from 1 to 30 days per treatment. If necessary, the same treatment can be repeated after a titration period.
  • the dose is the same as that of a human per kg, or the above dose is converted, for example, by the volume ratio (for example, average value) of the target animal and the organ (heart, etc.) of the human One dose can be administered.
  • the strain according to one aspect can increase the expression of skin moisturizing and skin barrier-related hyaluronic acid synthase 3 (Hyaluronan synthase 3, HAS3, filagrin) and regenerate damaged cells.
  • the strain can increase the expression of COL1A1 reduced by ultraviolet light, and decrease the increased expression of MMP-1. That is, the increase in markers such as HAS3 and filaggrin means that filaggrin, a precursor of NMF, and intercellular lipids are normalized in keratinocytes through the formation of a normal stratum corneum. Therefore, to prevent skin aging and maintain skin health from external environmental changes such as atmospheric drying, ultraviolet rays and various pollutants, to strengthen the skin barrier to prevent moisture loss in the skin tissue while promoting the maintenance of moisture in the skin tissue It can form a stratum corneum.
  • the strain promotes the proliferation of damaged fibroblasts and extracellular matrix to regenerate the skin and restore the epithelial layer, so it is useful for skin regeneration or wound healing.
  • Another aspect provides an oral composition
  • an oral composition comprising the strain, a lysate thereof, a culture solution, an extract of the culture solution, or a mixture thereof as an active ingredient.
  • Specific details of the strain are the same as described above.
  • the strain reduces the concentrations of volatile sulfur compounds (VSC), methyl mercaptan and dimethyl sulfide, which are volatile sulfur compounds (VSC) generated by bad breath-inducing microorganisms, and it was confirmed that the amount of oral microorganisms was reduced.
  • the hydrogen sulfide (Hydrogen sulfide, H 2 S) may cause bad breath due to oral contamination and diseases related to mental and physiological causes.
  • the methyl mercaptan (Methyl mercaptan, CH 3 SH) may cause bad breath due to oral diseases (periodontal disease).
  • dimethyl sulfide Dimethyl sulfide, (CH 3 ) 2 S) may cause transient bad breath due to food metabolism.
  • halitosis is a problem experienced in daily life, and 90% of it is derived from periodontitis, tongue discoloration, poor oral hygiene, and inadequate prosthetics in the oral cavity, and the remaining 10% is gastrointestinal disease, carcinoma, diabetes, liver failure, renal failure and from the same systemic disease. More than 600 types of bacteria live in the oral cavity, including disease-causing bacteria.
  • the strain according to an aspect may have an effect of improving oral contamination and various diseases, or bad breath caused by food metabolism, reducing plaque, reducing tartar, reducing dry mouth, or inhibiting oral bacteria.
  • the oral composition may be formulated as a mouthwash, mouthwash, toothpaste, oral spray, denture cleaner, toothpaste whitening agent, and the like.
  • Another aspect provides a pharmaceutical composition for preventing or treating oral diseases comprising the strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof as an active ingredient.
  • Another aspect provides a method of treating an oral disease comprising administering to a subject in need thereof an effective amount of the composition.
  • Another aspect provides a health functional food composition for preventing or improving oral diseases comprising the strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof as an active ingredient. Specific details of the strain are the same as described above.
  • the oral disease is, for example, invasive periodontitis, juvenile periodontitis, acute or chronic periodontitis, alveolar bone destruction, chronic or intractable periodontitis, gingivitis, ulcerative gingivitis, acute paronychia, acute lumpy gingivitis, hormonal periodontitis, complex infection, apical It may be a disease, periodontal abscess, dental caries, apical abscess, pulpitis, and the like.
  • composition containing the strain in one embodiment, after 2 weeks of use of the composition containing the strain, representative oral microorganisms Tannerella forsythia , Fusobacterium nucleatum , Prevotella nigrescens ( Prevotella ) nigrescens ), Streptococcus mitis and Streptococcus mutans were significantly reduced compared to before use, and it was confirmed that no adverse reactions due to long-term use were observed.
  • the strain according to one aspect has excellent oral microbial inhibitory effect as well as excellent stability for the human oral cavity, so even if used for a long period of time, it does not affect the resistance of bacteria in the oral cavity. effective in treatment.
  • Weissella sibaira strain according to an aspect, it can be usefully used for preventing, improving or treating skin-related conditions. In addition, it can be usefully used for the prevention, improvement or treatment of oral diseases by reducing the sulfur compounds causing bad breath and inhibiting oral microorganisms.
  • 1 is a graph showing the effect of a strain according to an embodiment on the expression of HAS3.
  • FIG. 2 is a graph showing the effect of a strain according to an embodiment on the expression of filaggrin.
  • FIG. 3 is a graph showing the effect of a strain according to an embodiment on skin regeneration or wound recovery.
  • FIG. 4 is a photograph showing the effect of a strain according to an embodiment on skin regeneration or wound recovery.
  • [Correction by Rule 91 24.02.2022] 5 is a graph showing the effect of a strain according to an embodiment on the expression of MMP-1.
  • 6A is a graph comparing changes in the concentration of hydrogen sulfide before, immediately after, and after 2 weeks of use of the composition according to an aspect.
  • 6B is a graph comparing changes in the concentration of methyl mercaptan before, immediately after, and 2 weeks after use of the composition according to an aspect.
  • 6c is a graph comparing changes in the concentration of dimethyl sulfide before, immediately after, and after 2 weeks of use of the composition according to an aspect.
  • Figure 7a is a graph comparing the amounts of bacteria Actinomycetem comitans ( Aa ) and gingivalis ( Pg ) before and 2 weeks after use of the composition according to an aspect.
  • Figure 7b is a graph comparing the amounts of bacteria forsythia ( Tf ) and bacteria denticola ( Td ) before and 2 weeks after use of the composition according to an aspect.
  • Figure 7c is a graph comparing the amounts of nucleatum bacteria ( Fn ) and intermediary bacteria ( Pi ) before and 2 weeks after use of the composition according to an aspect.
  • Figure 7d is a graph comparing the amount of bacteria nigrescens ( Pn ) and Mytisbacterium ( Smi ) before and 2 weeks after use of the composition according to an aspect.
  • Figure 7e is a graph comparing the amounts of bacteria mutans ( Smu ) and casei bacteria ( Lc ) before and 2 weeks after use of the composition according to an aspect.
  • 7f is a graph comparing the total amount of microorganisms before and 2 weeks after use of the composition according to an aspect.
  • FIG. 8 is a graph analyzing the results of a questionnaire evaluation regarding the efficacy and usability of a composition according to an aspect.
  • Samples collected from human oral saliva and kimchi were inoculated into a liquid or solid medium (Becton Dickinson, Cockeysville, MD) containing MRS. After inoculation, 100 colonies formed after culturing for 48 hours in an incubator at 28°C were purely separated and cultured, and cultured in an incubator at 28°C for 48 hours. The cultured colonies were subjected to 16s rRNA gene sequence identification.
  • the primers used were designed to amplify in response to bacteria only (27F, 5'-AGAGTTTGATCCTGGCTCAG-3'; 1492R, 5'-GGTTACCTTGTTACGACTT-3).
  • PCR amplification was carried out in 30 cycles of 95°C for 1 minute, 55°C for 1 minute, 75°C for 1 minute and 30 seconds, and finally, after treatment at 72°C for 8 minutes, it was stored at 4°C.
  • DNA sequences of the isolated and cultured species were determined using ABI-3730XL (ABI, USA). The nucleotide sequence of the 16S rRNA region determined from the isolated and cultured microbial colonies was compared with other strains registered in the BLAST program provided on the website of the National Center for Biotechnology Information (NCBI), and the homology of 97% or less was found.
  • NCBI National Center for Biotechnology Information
  • CXO-1 novel microorganism with 97% homology or less Weissella cibaria strain
  • the selected CXO-1 strain was deposited with the Korea Microbial Conservation Center on December 3, 2020 and was given an accession number KCCM12889P, and the CXO-1 strain has a 16s rRNA sequence of SEQ ID NO: 1 (complementary DNA).
  • HaCaT cells which are human keratinocytes, were cultured in DMEM medium (Dulbecco'smodified Eagle's Medium, Gibco 1210-0038) containing 10% fetal bovine serum, and all cultures were cultured at 37°C 5°C. % CO 2 in an incubator.
  • the cultured cell line was treated with the culture solution of the CXO-1 strain (1%, 10% (w/w)) and further cultured for 24 hours.
  • RNA was isolated from the cells of each sample using trizol (RNA iso, DAKARA, Japan), and RNA was quantified at 260 nm with nanodrops, and cDNA was synthesized in an amplifier using 2 ⁇ g of RNA each. (C1000 Thermal Cycler, Bio-Rad, USA).
  • the target gene, HAS3 (hyaluronic acid synthase), a factor related to skin barrier strengthening and moisturizing, and primers and cDNA for filaggrin were added together with a real-time PCR machine (Step One Plus, Real-time polymerase chain reaction was performed in Applied Biosystems, USA).
  • the expression level of the gene was finally analyzed through correction for the ⁇ -actin gene, and the results are shown in FIGS. 1 and 2 .
  • 1 is a graph showing the effect of a strain according to an embodiment on the expression of HAS3.
  • FIG. 2 is a graph showing the effect of a strain according to an embodiment on the expression of filaggrin.
  • the strain according to one embodiment restores the damaged skin barrier by significantly increasing the expression of HAS3 and filaggrin in a concentration-dependent manner, and strengthens the skin barrier to increase skin defense. can give In addition, it is excellent in moisturizing the skin and is effective in preventing or treating moisture-related diseases.
  • Example 2 In order to evaluate the skin regeneration and wound healing efficacy of the CXO-1 strain isolated in Example 1, a cell migration assay was performed. Specifically, Hs68 human fibroblasts were seeded in 96-well ImageLock plate (No. 4379; Essen BioScience) at 2 ⁇ 10 4 cells/well. Experiments were conducted at 90% cell confluent. On the second day, WoundMaker (Essen BioScience, USA) was washed with 45 ml of 70% ethanol solution for 5 minutes, and then washed with 45 ml of distilled water for 5 minutes. All cell layers in each well of the plate were scratched using the washed WoundMakerTM. Then, the medium was removed and washed with PBS.
  • WoundMaker Essen BioScience, USA
  • each test substance was treated.
  • the degree of wound recovery was measured and graphed at 2-hour intervals for 8 hours.
  • the degree of wound recovery was visually confirmed by checking the photograph of the cell measured with a microscope in the device.
  • FIG. 3 is a graph showing the effect of a strain according to an embodiment on skin regeneration or wound recovery.
  • FIG. 4 is a photograph showing the effect of a strain according to an embodiment on skin regeneration or wound recovery.
  • the strain according to one embodiment promotes skin regeneration by activating cells in the wound site, and thus can be used as a material for skin regeneration or wound healing.
  • Example 1 The effect of the CXO-1 strain culture medium isolated in Example 1 on factors related to aging skin aging and wrinkle generation by ultraviolet (UV) irradiation was analyzed.
  • a human dermal fibroblast (Hs68) cell line was seeded in a 6-well plate at 3.5x10 5 cells/well, and then cultured in an incubator at 37° C. and 5% CO 2 condition for 24 hours. After removing the medium and adding DPBS, 20 mJ/cm2 of UVB was irradiated or not. Immediately after UVB irradiation, after removing DPBS and replacing it with a medium without FBS, the culture solution of the CXO-1 strain was treated by concentration (1% (w/w), 10% (w/w)) and added for 24 hours cultured. As a negative control group, UV-treated and strain culture untreated groups were used.
  • Figure 5 is a graph showing the effect of the strain according to one embodiment on the expression of COL1A1
  • Figure 5b is a graph showing the effect of the strain according to one embodiment on the expression of MMP-1.
  • the strain according to one aspect has an excellent effect in preventing skin aging, such as reducing elasticity and inhibiting wrinkle production, by restoring collagen destroyed by ultraviolet rays by ultraviolet rays, and inhibiting the production of MMP-1 increased by ultraviolet rays .
  • VSC Volatile Sulfur Compound
  • 6A to 6C are graphs comparing changes in the concentration of volatile sulfur compounds (hydrogen sulfide, methyl mercaptan and dimethyl sulfide) before, immediately after, and after 2 weeks of use of the composition according to an aspect.
  • the concentration of volatile sulfur compounds which is a parameter for bad breath, significantly decreased immediately after use and 2 weeks after use compared to before use (p ⁇ 0.05).
  • the rate of change of volatile sulfur compounds is 89.41% and 87.80% for hydrogen sulfide, 68.46%, and 82.05% for methyl mercaptan, and 98.12% and 90.43% for dimethyl sulfide. After 2 weeks, it was confirmed that each concentration decreased by 80% or more.
  • composition according to an aspect is effective in improving bad breath by significantly reducing the concentration of volatile sulfur compounds.
  • the sample container was provided from the oral pathogenic microorganism test requesting institution (DENOMICS, Korea). After collecting a volunteer's saliva sample, changes in oral microorganisms and total amount of microorganisms in Table 2 below were analyzed using Oral Bacteria Dignosis (OBD) analysis service, and the results are shown in Table 3 below.
  • OBD Oral Bacteria Dignosis
  • 7A to 7E are graphs comparing the amount of microorganisms described in Table 2 before and after 2 weeks of use of a composition according to an aspect.
  • 7f is a graph comparing the total amount of microorganisms before and 2 weeks after use of the composition according to an aspect.
  • composition according to one aspect can prevent or treat oral diseases such as periodontitis, gingivitis, apical disease, dental caries, pulpitis, and the like, as well as prevent tartar formation by significantly reducing oral microorganisms.
  • oral diseases such as periodontitis, gingivitis, apical disease, dental caries, pulpitis, and the like, as well as prevent tartar formation by significantly reducing oral microorganisms.
  • the efficacy of improving bad breath, reducing plaque, and reducing dry mouth was evaluated according to the evaluation criteria 1 of Table 4 below, and the results were obtained. It is shown in Table 5 below.
  • usability such as freshness, stimulation, flavor and taste were evaluated, and the results are shown in Table 6 below.
  • FIG. 8 is a graph analyzing the results of a questionnaire evaluation regarding the efficacy and usability of a composition according to an aspect.
  • the composition containing the strain culture solution of Example 1 was evaluated to have excellent effects of improving bad breath, reducing plaque, and reducing dry mouth.
  • the composition was evaluated as being refreshing and having no irritation, so that the feeling of use was excellent overall.
  • composition according to an aspect may provide an oral care product having excellent feeling of use as well as functionalities such as improving bad breath, reducing plaque, and reducing dry mouth.
  • the 20 volunteers were allowed to use the composition containing the culture medium of Example 1 for 2 weeks, and then the test site of the volunteers was observed. Thereafter, the state of the test site was checked, recorded, and evaluated through Q&A. In the event of an adverse reaction, an adverse reaction report was prepared, and an expert judged the relationship between the adverse reaction and the composition.
  • the composition according to one aspect has an advantage in that it has excellent oral microbial inhibitory effect as well as excellent stability to the human oral cavity, so that even when used for a long period of time, it does not affect the resistance of bacteria in the oral cavity.

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Abstract

La présente invention concerne une souche de Weissella cibaria et son utilisation. La souche peut être utilement utilisée pour la prévention, l'amélioration ou le traitement des affections liées à la peau, et réduit les composés sulfurés provoquant l'halitose et inhibe les micro-organismes buccaux, et peut donc être utilement utilisée pour la prévention, l'amélioration ou le traitement des maladies buccales.
PCT/KR2021/020031 2020-12-28 2021-12-28 Souche de weissella cibaria et son utilisation WO2022145960A1 (fr)

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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20060056996A (ko) * 2004-08-25 2006-05-25 전남대학교산학협력단 구취를 억제하는 유산균
KR20150031922A (ko) * 2013-09-17 2015-03-25 충북대학교 산학협력단 신규한 와이셀라 시바리아 jw15 균주 및 이의 용도
KR20160035219A (ko) * 2014-09-23 2016-03-31 한국 한의학 연구원 틴달화 유산균 사균체를 유효성분으로 포함하는 피부 보습 또는 주름개선용 조성물
KR101667496B1 (ko) * 2015-10-15 2016-10-18 한국식품연구원 김치 유산균 와이셀라 시바리아(Weissella cibaria) WIKIM28을 유효성분으로 포함하는 아토피 피부염 치료용 약학 조성물
KR20180059728A (ko) * 2018-04-06 2018-06-05 (주)앰틱스바이오 신규한 와이셀라 시바리아 균주 및 이의 용도
KR20200070989A (ko) * 2018-12-10 2020-06-18 주식회사 엠디헬스케어 웨이셀라 속 세균 유래 나노소포 및 이의 용도

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20150065170A (ko) * 2012-08-07 2015-06-12 탑제닉스, 인크. 관심있는 화합물을 발현하는 형질전환된 박테리아를 포함하는 국소 조성물
JP6438668B2 (ja) 2014-03-28 2018-12-19 株式会社ロッテ ワイセラ属乳酸菌
KR101873393B1 (ko) * 2016-12-21 2018-07-02 창원대학교 산학협력단 신규한 웨이셀라 시바리아 ｂｃｎｕ ３００３ 균주
EP3428286A1 (fr) 2017-07-13 2019-01-16 Université de Bordeaux Test prédictifs de réponse anti-tnf alpha chez des patients atteints d'une maladie inflammatoire
KR102404383B1 (ko) * 2020-05-27 2022-06-07 주식회사 피토메카 와이셀라 시바리아 세포 용해물을 포함하는 피부 노화 방지 또는 주름 개선용 화장료 조성물

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20060056996A (ko) * 2004-08-25 2006-05-25 전남대학교산학협력단 구취를 억제하는 유산균
KR20150031922A (ko) * 2013-09-17 2015-03-25 충북대학교 산학협력단 신규한 와이셀라 시바리아 jw15 균주 및 이의 용도
KR20160035219A (ko) * 2014-09-23 2016-03-31 한국 한의학 연구원 틴달화 유산균 사균체를 유효성분으로 포함하는 피부 보습 또는 주름개선용 조성물
KR101667496B1 (ko) * 2015-10-15 2016-10-18 한국식품연구원 김치 유산균 와이셀라 시바리아(Weissella cibaria) WIKIM28을 유효성분으로 포함하는 아토피 피부염 치료용 약학 조성물
KR20180059728A (ko) * 2018-04-06 2018-06-05 (주)앰틱스바이오 신규한 와이셀라 시바리아 균주 및 이의 용도
KR20200070989A (ko) * 2018-12-10 2020-06-18 주식회사 엠디헬스케어 웨이셀라 속 세균 유래 나노소포 및 이의 용도

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