WO2022145960A9 - Souche de weissella cibaria et son utilisation - Google Patents

Souche de weissella cibaria et son utilisation Download PDF

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WO2022145960A9
WO2022145960A9 PCT/KR2021/020031 KR2021020031W WO2022145960A9 WO 2022145960 A9 WO2022145960 A9 WO 2022145960A9 KR 2021020031 W KR2021020031 W KR 2021020031W WO 2022145960 A9 WO2022145960 A9 WO 2022145960A9
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Prior art keywords
strain
culture medium
skin
oral
composition
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PCT/KR2021/020031
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English (en)
Korean (ko)
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WO2022145960A1 (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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    • 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
    • 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
    • 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 ecosystem of the skin provides various types of habitats for microorganisms, and a wide range of microorganisms live there. They form a symbiotic relationship with the host, human, and are known to have many positive effects on the host.
  • the skin constitutes various types of habitats, such as indentations 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 risk factors and toxic substances from the outside.
  • the skin becomes a point of contact with the external environment and is also a gathering place for various microorganisms (fungi, bacteria, viruses, and small larvae). According to the selection of physical and chemical functions, microorganisms adapt to specialized niches to prepare habitats.
  • the skin is cold, has an acidic nature, and remains dry. Structurally, the epidermis forms the 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 form called 'brick and mortar structure', and the skin tissue undergoes a continuous self-healing process, and the scales that have passed through the end of the differentiation process are constantly eliminated from the skin tissue.
  • these metal salts or disinfectants inhibit not only anaerobic bacteria that cause bad breath, but also other microorganisms in the oral cavity, and since these substances are not swallowed into the esophagus, they do not reach the back of the tongue, which is an important place in the occurrence of bad breath, but mainly in the front of the mouth. Gargle and spit it out of your mouth.
  • these substances are diluted by saliva and swallowed into the esophagus together with saliva, and the effect in the oral cavity is short, about 20 minutes to 2 hours, after which microorganisms proliferate again in the oral cavity, resulting in bad breath.
  • Lactic acid bacteria refers to bacteria that produce lactic acid as a final metabolite by fermenting carbohydrates, and lactic acid bacteria exist in the oral cavity and digestive tract of humans and animals.
  • the lactic acid bacteria are used in the manufacturing process of fermented foods such as kimchi or yogurt.
  • Lactic acid bacteria used in food include Enterococcus genus, Lactobacillus genus, Lactococcus genus, Leuconostoc ( Leuconostoc ), Streptococcus ( Streptococcus ), Weissella ( Weissella ), 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 diluted by saliva, such as metal salts or disinfectants, and are swallowed directly into the esophagus, making it difficult to remain in the oral cavity.
  • lactic acid bacteria such as Lactobacillus acidophilus , Lactobacillus casei , and Lactobacillus salivarius can produce strong acids to inhibit the growth of anaerobic bacteria in a test tube, but are In the oral cavity, the buffering action of saliva neutralizes the strong acid produced by lactic acid bacteria, making it difficult to inhibit the growth of other bacteria.
  • strong acids cause dental caries, long-term administration of these bacteria to the oral cavity is not good for oral hygiene. Therefore, there is a need to develop microorganisms to reduce bad breath.
  • One aspect is to provide a Weissella cibaria CXO-1 strain belonging to the Weissella sp .
  • Another aspect is to provide a lysate or culture medium of the strain.
  • Another aspect is to provide a cosmetic composition
  • a cosmetic composition comprising the strain, a lysate thereof, a culture medium, an extract of the culture medium, or a mixture thereof.
  • Another aspect is to provide a pharmaceutical composition for wound healing comprising the strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof.
  • Another aspect is to provide an oral composition
  • an oral composition comprising the strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof.
  • Another aspect is to provide 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.
  • Another aspect is to provide a wound treatment method comprising the step of administering to a subject a composition containing the strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof.
  • Another aspect is to provide a method for treating oral diseases comprising the step of administering to a subject a composition comprising the strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof.
  • Another aspect is to provide a composition containing the strain, its lysate, culture medium, culture medium extract or a mixture thereof for wound treatment, prevention or treatment of oral diseases.
  • the bicellar Cibaira ( Weissellacibaria )
  • the strain may be a strain deposited with accession number KCCM12889P.
  • the Weissella cibira ( Weissellacibaria ) strain may be a strain containing the 16s rRNA of SEQ ID NO: 1.
  • the strain increases the expression of anti-aging and anti-wrinkle factors, such as COL1A1 or reduces the expression of MMP-1, and hyaluronan synthase 3 (Hyaluronan synthase 3), a skin moisturizing and skin barrier-related factor. 3, HSA3) or filaggrin expression was confirmed to increase.
  • the strain according to one aspect may have a skin beauty improvement effect, for example, skin aging inhibition, skin barrier enhancement, skin wrinkle inhibition, or skin moisturizing effect.
  • Another aspect provides a lysate of the strain, a culture medium, or an extract of the culture medium. Details of the strain are as described above.
  • the term "culture medium” may be used interchangeably with “culture supernatant”, “conditioned culture medium” or “conditioned medium”, and may supply nutrients so that the Visella cibaira strain can grow and survive in vitro. It may mean the entire medium including the strain, its metabolites, extra nutrients, etc. obtained by culturing the strain for a certain period of time in a medium that can be used.
  • the culture solution may mean a culture solution obtained by removing the cells from the cell culture solution obtained by culturing the strain.
  • the liquid from which the cells are removed from the culture solution is also called “supernatant". It can be obtained by removing the precipitate and taking only the upper liquid.
  • the "cell” refers to the “strain” itself of the present invention, and includes the “strain” itself separated and selected from skin samples, etc., or the “strain” separated from the culture solution by culturing the "strain".
  • the cells can be obtained by centrifuging the culture solution and taking the part that has sunk in the lower layer, or it can be obtained by leaving it for a certain period of time and then removing the upper liquid as it sinks to the lower layer of the culture medium by gravity.
  • the culture solution may include a culture solution itself obtained by culturing the strain, a concentrate thereof, a lyophilized product, or a culture supernatant obtained by removing the strain from the culture solution, a concentrate thereof, or a lyophilized product.
  • the culture medium is obtained by culturing the Vicella cibira strain in a medium (eg, R2A medium or TSA medium) at any temperature above 10 ° C or below 40 ° C for a certain period of time, for example, 4 to 50 hours can
  • the culture supernatant of the strain can be obtained by centrifuging or filtering the strain culture medium to remove the strain.
  • the concentrate may be obtained by concentrating the supernatant obtained after filtering the strain culture medium itself, or the culture medium using a centrifugal separation or filter.
  • lysate may mean a product obtained by disrupting the cell wall of the strain itself by chemical or physical force.
  • culture medium extract refers to an extract obtained from the culture medium or a concentrate thereof, and includes an extract, a diluent or concentrate of the extract, a dried product obtained by drying the extract, or a crude or purified product thereof, or a fraction obtained by fractionating the same. can do.
  • Another aspect provides the use of the strain, a lysate of the strain, a culture medium, or an extract of the culture medium. Specifically, it provides a composition containing the extract of the Visella cibira strain, its lysate, culture medium or its culture medium.
  • the use of the strain may include improvement of skin conditions, improvement of skin beauty, prevention, improvement or treatment of skin diseases.
  • the skin condition improvement or skin beauty improvement may be suppression or improvement of skin aging, anti-wrinkle, skin elasticity, skin regeneration, skin barrier strengthening, skin moisturizing or wound improvement.
  • skin aging refers to both tangible and intangible changes that appear in the skin with age, such as thinning of the epidermis, number of cells or blood vessels in the dermis, DNA damage repair ability, cell replacement cycle , wound healing, skin barrier function, epidermal moisture retention, sweat secretion, sebum secretion, vitamin D production, physical damage defense, chemical substance elimination ability, immune response, sensory function, and decrease in body temperature control.
  • the strain or its culture medium may be used to improve skin aging caused by extrinsic factors or endogenous factors.
  • the extrinsic factor refers to various external factors, such as ultraviolet rays (light), and the endogenous 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 is specifically a natural aging phenomenon that occurs as the proliferation of skin cells decreases with age as well as premature aging symptoms induced by external stimuli such as ultraviolet rays, pollution, cigarette smoke, chemicals, etc. It is a concept that includes wrinkles, loss of elasticity, skin sagging and dryness. In addition, wrinkles include stimulation caused by changes in internal and external factors that cause wrinkles by changing components constituting skin tissue.
  • the aging may be photoaging.
  • photoaging is a phenomenon caused by external environmental factors, and the most representative factor is ultraviolet rays.
  • Ultraviolet rays cause damage to biocomponents such as activation of proteolytic enzymes, chain scission of matrix proteins, and abnormal cross-linking, and repetition of these mechanisms results in apparent skin aging.
  • wrinkle refers to a state in which elasticity of the skin is lost and loosened, and for example, the skin may be folded.
  • prevention or improvement of skin wrinkles may mean any action of preventing or improving wrinkles by suppressing the expression of wrinkle-related factors or increasing the total amount of collagen.
  • the “strengthen the skin barrier” may refer to any action that enhances the function of the skin barrier, which is located at the outermost layer of the skin and prevents loss of moisture and nutrients.
  • damaged skin barrier function may refer to all changes in the skin due to a decrease or damage to the function of the skin barrier. Examples include increased skin wrinkling, dryness, dermatitis, atopic dermatitis, allergic dermatitis, acne, and the like.
  • the "moisturizing skin” may refer to any action that maintains skin moisture or prevents moisture loss.
  • wound is also referred to as “wound”, and means damage to a living body.
  • the loss may be caused by external factors such as physical damage, stimulation by radiation, chemicals, etc., proliferation of microorganisms, or internal factors such as stress.
  • the wounds may include all kinds of wounds such as cut wounds, puncture wounds, crack wounds, acne wounds, fissure wounds, thrush wounds, and wound wounds.
  • Wound improvement may mean any action that fades or reduces the extent of a wound on the face or body, and "wound healing” may mean a series of biological reactions to recover a wound.
  • the strain may be used together with other strains belonging to the genus Bycella having a skin improvement effect to exhibit a synergistic effect.
  • Strains belonging to the genus Bisella are, for example, Bisella confuse ( Weissella confuse ), It may be Weissella uvarum and the like.
  • the composition is 0.001% to 80% by weight, for example, 0.01% to 60% by weight, 0.01% to 40% by weight, 0.01% to 30% by weight, 0.01% to 20% by weight, 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%, 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 a strain, a lysate thereof, a culture medium, or an extract of a culture medium thereof.
  • the composition may be a cosmetic composition.
  • the cosmetic composition may have, for example, softening lotion, nutrient lotion, massage cream, nutrient cream, essence, pack, gel, ampoule, or skin-adhesive cosmetic formulation.
  • ingredients included in the cosmetic composition may include ingredients commonly used in cosmetic compositions other than the composition as active ingredients, for example, conventional adjuvants and carriers such as stabilizers, solubilizers, vitamins, pigments and flavors. can include
  • the composition may be a composition for external application for skin.
  • the external skin preparation may be a cream, gel, ointment, skin emulsifier, skin suspension, transdermal delivery patch, drug-containing bandage, lotion, or a combination thereof.
  • the external skin preparation is a component usually used in external preparations for skin such as cosmetics or pharmaceuticals, for example, water-based components, oil-based components, powder components, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, and fragrances. , colorants, various skin nutrients, or combinations thereof and may be suitably blended as needed.
  • the external skin preparations include metal sequestering agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid, caffeine, tannin, bellapamil, licorice extract, glabridin, and calin.
  • Hot-water extracts of fruits, various herbal medicines, tocopherol acetate, glycyrrhizic acid, tranexamic acid and its derivatives or salts and other drugs, vitamin C, magnesium ascorbate phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, Sugars, such as trehalose, etc. can also be mix
  • the composition may be a pharmaceutical composition.
  • the pharmaceutical composition may additionally include a pharmaceutically acceptable diluent or carrier.
  • the diluent may be lactose, corn starch, soybean oil, microcrystalline cellulose, or mannitol, and magnesium stearate, talc, or a combination thereof as a lubricant.
  • 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 calcium carboxymethylcellulose, sodium starch glycolate, 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 for oral or parenteral administration.
  • Oral dosage forms may be granules, powders, solutions, tablets, capsules, dry syrups, or combinations thereof.
  • Parenteral dosage forms may be injections.
  • the composition may be a health functional food composition.
  • the health functional food composition may be used alone or in combination with the strain or its culture medium or other food or food component, 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 (prevention, 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.
  • beverage compositions may contain various flavoring agents or natural carbohydrates as additional components, like conventional beverages.
  • 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 thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used.
  • the health food composition may also contain nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonated beverages. carbonation agent used, or a combination thereof.
  • the health functional food composition may also contain natural fruit juice, fruit juice beverages, fruit flesh for preparing vegetable beverages, or a combination thereof.
  • Another aspect provides a method for preventing, improving or treating a skin condition in a subject comprising treating or administering to a subject in need thereof an effective amount of the composition. Details of the skin condition are as described above.
  • Another aspect provides a pharmaceutical composition for treating wounds containing the strain, its lysate, culture medium, an extract of the culture medium, or a mixture thereof as an active ingredient.
  • Another aspect provides a composition for external application 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 wound treatment method comprising administering an effective amount of the strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof to a subject in need thereof. Details of the wound are as described above.
  • administering As used herein, the terms “administering,” “introducing,” and “implanting” are used interchangeably and according to one embodiment into a subject by a method or route that results in at least partial localization of a composition to a desired site. It may mean the arrangement of the composition according to one embodiment of.
  • Administration may be administered by a method known in the art.
  • Administration can be administered directly to a subject by any means, for example, by routes such as intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal or subcutaneous administration.
  • routes such as intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal or subcutaneous administration.
  • the administration may be administered 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 a subject in need of skin beauty improvement, for example, skin moisturizing, skin barrier strengthening, and skin wrinkle improvement effects.
  • the administration is 0.1 mg to 1,000 mg per day of the composition according to one embodiment, 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 can Dosage can be appropriately adjusted in consideration of these factors.
  • the number of administrations can be once a day or twice or more within the range of clinically acceptable side effects, and the administration site can be administered to one or more than two sites, daily or every 2 to 5 days, total
  • the number of administration days may be administered from 1 day to 30 days per treatment. If necessary, the same treatment can be repeated after a titration period.
  • the same dosage per kg as for humans is used, or the above dosage is converted by the volume ratio (eg, average value) of the organ (heart, etc.) between the target animal and the human.
  • a single 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 reduce 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, in order to prevent skin aging and maintain skin health from external environmental changes such as air dryness, ultraviolet rays and various pollutants, to strengthen the skin barrier to prevent loss of moisture in skin tissue while promoting moisture retention in skin tissue stratum corneum can be formed.
  • 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, its lysate, culture medium, an extract of the culture medium, or a mixture thereof as an active ingredient. Details of the strain are as described above.
  • the strain reduces the concentration of hydrogen sulfide, methylmercaptan, and dimethylsulfide, which are volatile sulfur compounds (VSC) generated by bad breath-inducing microorganisms, and reduces the amount of oral microorganisms.
  • the hydrogen sulfide (H 2 S) may cause bad breath due to diseases related to oral contamination and mental and physiological causes.
  • the methyl mercaptan (CH 3 SH) may cause bad breath due to oral disease (periodontal disease).
  • dimethyl sulfide ((CH 3 ) 2 S) may cause transient bad breath due to food metabolism.
  • halitosis is a problem experienced in daily life, 90% of which is derived from periodontitis, tongue formation, poor oral hygiene, and improper prosthesis in the oral cavity, and the remaining 10% is caused by gastrointestinal diseases, carcinoma, diabetes, liver failure, renal failure and stem from the same systemic disease. More than 600 species of bacteria live in the oral cavity, including disease-causing bacteria. Oral bacteria, bacteria living in plaque and on the tongue can cause bad breath by producing an unpleasant odor.
  • the strain according to one aspect may have an effect of improving oral pollution and various diseases, or bad breath caused by food metabolism, reducing plaque, reducing calculus, reducing oral dryness, 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, an extract of the culture medium, or a mixture thereof as an active ingredient.
  • Another aspect provides a method of treating oral disease comprising administering an effective amount of the composition to a subject in need thereof.
  • 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. Details of the strain are as described above.
  • the oral disease is, for example, invasive periodontitis, juvenile periodontitis, acute or chronic periodontitis, alveolar bone destruction, chronic or refractory periodontitis, gingivitis, ulcerative gingivitis, acute perironditis, acute massive 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 after 2 weeks of using the composition containing the strain, representative oral microorganisms Tannerella Forsythia , Fusobacterium nucleatum , Prevotella nigrescens bacteria ( Prevotella nigrescens ), Streptococcus mitis bacteria ( Streptococcus mitis ) and Streptococcus mutans bacteria ( Streptococcus mutans ) were significantly reduced compared to before use, and it was confirmed that no adverse reactions were observed due to long-term use.
  • the strain according to one aspect is not only excellent in inhibiting oral microorganisms, but also has excellent stability in the oral cavity of the human body, and does not affect the resistance of bacteria in the oral cavity even when used for a long time, preventing or preventing oral diseases such as periodontitis, gingivitis, dental caries, etc. effective in treatment
  • Visella cibira strain according to one aspect, it can be usefully used for the prevention, improvement or treatment of skin-related conditions. In addition, it can be usefully used for preventing, improving or treating oral diseases by reducing sulfur compounds that cause bad breath and inhibiting oral microorganisms.
  • 1 is a graph showing the effect of a strain according to one embodiment on the expression of HAS3.
  • Figure 2 is a graph showing the effect of the strain on the expression of filaggrin according to one embodiment.
  • Figure 3 is a graph showing the effect of the strain on skin regeneration or wound recovery according to one embodiment.
  • Figure 4 is a photograph showing the effect of the strain according to one embodiment on skin regeneration or wound recovery.
  • Figure 5 is a graph showing the effect of the strain on the expression of MMP-1 according to one embodiment.
  • Figure 6a is a graph comparing changes in the concentration of hydrogen sulfide before, immediately after, and after 2 weeks of use of a composition according to one aspect.
  • 6B is a graph comparing changes in the concentration of methylmercaptan before, immediately after, and after 2 weeks of use of a composition according to one aspect.
  • Figure 6c is a graph comparing the change in the concentration of dimethyl sulfide before, immediately after, and after 2 weeks of use of a composition according to one aspect.
  • Figure 7a is a graph comparing the amount of Actinomycetemcomitans bacteria ( Aa ) and gingivalis bacteria ( Pg ) before and after 2 weeks of use of the composition according to one aspect.
  • Figure 7b is a graph comparing the amount of bacteria Forsythia ( Tf ) and bacteria Denticola ( Td ) before and after 2 weeks of use of the composition according to one aspect.
  • Figure 7c is a graph comparing the amounts of nucleatum bacteria ( Fn ) and intermedia bacteria ( Pi ) before and after 2 weeks of use of the composition according to one aspect.
  • Figure 7d is a graph comparing the amount of nigrescens bacteria ( Pn ) and mytis bacteria ( Smi ) before and after 2 weeks of use of the composition according to one aspect.
  • Figure 7e is a graph comparing the amount of mutans bacteria ( Smu ) and casei bacteria ( Lc ) before and after 2 weeks of use of the composition according to one aspect.
  • Figure 7f is a graph comparing the total amount of microorganisms before and after 2 weeks of use of the composition according to one aspect.
  • PCR amplification was carried out in 30 cycles of 95 ° C for 1 minute, 55 ° C for 1 minute, and 75 ° C for 1 minute and 30 seconds, and finally treated at 72 ° C for 8 minutes and 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 colony was compared and analyzed with other strains registered in the BLAST program provided on the website of the National Center for Biotechnology Information (NCBI), and homology of 97% or less was obtained.
  • CXO-1 novel microbial strain of Weissella cibaria having 97% or less homology
  • the selected CXO-1 strain was deposited with the Korea Microorganism Conservation Center on December 3, 2020 and was given the accession number KCCM12889P, and the CXO-1 strain has a 16s rRNA sequence of SEQ ID NO: 1 (complementary DNA).
  • Example 1 The effect of the culture solution of the CXO-1 strain isolated in Example 1 on skin barrier strengthening and skin moisturizing activity was analyzed. Specifically, human keratinocyte HaCaT cells were cultured in DMEM medium (Dulbecco's modified Eagle's Medium, Gibco 1210-0038) containing 10% fetal bovine serum, and all cultures were carried out at 37 ° C 5 % CO 2 was performed in an incubator. The cultured cell line was treated with the culture medium (1%, 10% (w/w)) of the CXO-1 strain and further cultured for 24 hours.
  • DMEM medium Dulbecco's modified Eagle's Medium
  • 1 is a graph showing the effect of a strain according to one embodiment on the expression of HAS3.
  • Figure 2 is a graph showing the effect of the strain on the expression of filaggrin according to one embodiment.
  • 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.
  • it is excellent in moisturizing the skin and is effective in preventing or treating diseases related to moisturizing.
  • cell migration assay was performed. Specifically, Hs68 human fibroblasts were seeded in a 96-well ImageLockTM plate (No. 4379; Essen BioScience) at 2 ⁇ 10 4 cells/well. Experiments were conducted at 90% cell confluent. On the second day, the 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 of each well of the plate were scratched using the washed WoundMaker. Then, the medium was removed and washed with PBS.
  • each test substance was treated.
  • the degree of wound healing was measured at 2-hour intervals for 8 hours and graphed.
  • the degree of wound healing was visually confirmed by checking a photograph of cells measured under a microscope in the device.
  • Figure 3 is a graph showing the effect of the strain on skin regeneration or wound recovery according to one embodiment.
  • Figure 4 is a photograph showing the effect of the strain according to one embodiment on skin regeneration or wound recovery.
  • the strain according to one embodiment promotes skin regeneration by activating cells of a 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 solution isolated in Example 1 on factors related to aging skin aging and wrinkle formation by ultraviolet (UV) irradiation was analyzed.
  • human dermal fibroblast cell line Human dermal fibroblast, Hs68
  • Human dermal fibroblast, Hs68 was dispensed in a 6-well plate at 3.5x10 5 cells/well, and then cultured for 24 hours in an incubator at 37°C and 5% CO 2 conditions. Thereafter, the medium was removed, DPBS was added, and 20 mJ/cm 2 of UVB was irradiated or not irradiated. After removing DPBS immediately after UVB irradiation and changing to a medium without FBS, the culture medium of the CXO-1 strain was treated at different concentrations (1% (w/w), 10% (w/w)) and added for 24 hours cultured.
  • cybergreen SYBR Green supermix, Applied Biosystems, USA
  • a real-time PCR machine Step One Plus, Real-time polymerase chain reaction was performed at Applied Biosystems, USA.
  • Figure 5 is a graph showing the effect of the strain on the expression of COL1A1 according to one embodiment
  • Figure 5b is a graph showing the effect of the strain on the expression of MMP-1 according to one embodiment.
  • the strain according to one aspect recovers collagen destroyed by ultraviolet rays by ultraviolet rays and inhibits the production of MMP-1 increased by ultraviolet rays, thereby having an excellent effect in preventing skin aging such as reducing elasticity and suppressing wrinkle formation. .
  • VSC Volatile Sulfur Compound
  • 6a to 6c are graphs comparing concentration changes of volatile sulfur compounds (hydrogen sulfide, methyl mercaptan, and dimethyl sulfide) before, immediately after, and after 2 weeks of use of a composition according to one 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 was 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, respectively. After 2 weeks, it was confirmed that each concentration was reduced by more than 80%.
  • composition according to one aspect is effective in improving bad breath by significantly reducing the concentration of volatile sulfur compounds.
  • Example 2 After having the same subject as above use the composition containing the strain culture solution of Example 1 in the same way, a sample container was provided from the oral pathogenic microorganism test requesting institution (DENOMICS, Korea) before use and 2 weeks after use. After collecting saliva samples from volunteers, oral bacteria and total microorganisms in Table 2 were analyzed using OBD (Oral Bacteria Dignosis) 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 one aspect.
  • Figure 7f is a graph comparing the amount of total microorganisms before and after 2 weeks of use of a composition according to one 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 composition containing the strain culture solution of Example 1 was evaluated to have excellent effects on improving bad breath, reducing plaque and reducing dry mouth.
  • the composition was evaluated as being generally excellent in feeling of use because it was refreshing and non-irritating.
  • composition according to one aspect may provide an oral care product having functions such as improving bad breath, reducing plaque, and reducing oral dryness as well as providing an excellent feeling of use.
  • the 20 volunteers were allowed to use the composition containing the strain culture solution 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 question and answer. In the event of an adverse reaction, an adverse reaction report was prepared, and the relationship between the adverse reaction and the composition was determined by an expert.
  • the composition according to one aspect has an advantage of not only having an excellent effect of inhibiting oral microorganisms, but also having excellent stability in the oral cavity of the human body, so that the resistance of bacteria in the oral cavity is not affected even when used for a long period of time.

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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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WO2006022471A1 (fr) * 2004-08-25 2006-03-02 Jong Suk Oh Bacterie d'acide lactique arretant la mauvaise haleine
WO2014025938A1 (fr) * 2012-08-07 2014-02-13 TopGeniX, Inc. Composition topique comprenant des bactéries transformées exprimant un composé d'intérêt
KR101536672B1 (ko) * 2013-09-17 2015-07-24 충북대학교 산학협력단 신규한 와이셀라 시바리아 jw15 균주 및 이의 용도
JP6438668B2 (ja) * 2014-03-28 2018-12-19 株式会社ロッテ ワイセラ属乳酸菌
KR101693574B1 (ko) * 2014-09-23 2017-01-06 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
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
KR101869221B1 (ko) * 2018-04-06 2018-06-19 (주)앰틱스바이오 신규한 와이셀라 시바리아 균주 및 이의 용도
KR102356626B1 (ko) * 2018-12-10 2022-01-28 주식회사 엠디헬스케어 웨이셀라 속 세균 유래 나노소포 및 이의 용도
KR102404383B1 (ko) * 2020-05-27 2022-06-07 주식회사 피토메카 와이셀라 시바리아 세포 용해물을 포함하는 피부 노화 방지 또는 주름 개선용 화장료 조성물

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