WO2023018077A1 - Souche de microccocus cohnii et son utilisation pour améliorer l'état de la peau - Google Patents
Souche de microccocus cohnii et son utilisation pour améliorer l'état de la peau Download PDFInfo
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- WO2023018077A1 WO2023018077A1 PCT/KR2022/011230 KR2022011230W WO2023018077A1 WO 2023018077 A1 WO2023018077 A1 WO 2023018077A1 KR 2022011230 W KR2022011230 W KR 2022011230W WO 2023018077 A1 WO2023018077 A1 WO 2023018077A1
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- skin
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- micrococcus
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Classifications
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- 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
- C12N1/205—Bacterial isolates
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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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
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- 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
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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- A—HUMAN NECESSITIES
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- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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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
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- 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
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
Definitions
- the ecosystem of the skin provides various types of habitats for microorganisms, and a wide range of microorganisms live there.
- the human host forms a symbiotic relationship with them, and they 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.
- 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 retaining 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 differentiation process are constantly eliminated from the skin tissue.
- Probiotics is a collective term for microorganisms that have a beneficial effect on the human body, and refers to microorganisms that benefit the body. Most probiotics known to date are known as lactic acid bacteria. Probiotics have been reported to produce effective effects through various beneficial actions on the human body, but studies on the interaction between skin flora and skin are insufficient.
- One aspect is to provide novel Micrococcus cohnii strains.
- Another aspect is to provide a lysate of the strain, a culture medium, or an extract of the culture medium.
- Another aspect is to provide a composition
- a composition comprising an extract of the Micrococcus cohnii strain, its lysate, culture medium, or culture medium.
- Another aspect provides a method for preventing, ameliorating, or treating a condition in a subject in need thereof comprising administering to a subject in need thereof an effective amount of a composition described above.
- compositions comprising a Micrococcus cohnii strain belonging to the Micrococcus sp., a lysate thereof, a culture medium, or an extract of the culture medium for the manufacture of a cosmetic for improving skin conditions. to provide.
- compositions comprising a Micrococcus cohnii strain belonging to the Micrococcus sp., a lysate thereof, a culture medium, or an extract of the culture medium for the preparation of a therapeutic agent for skin inflammatory diseases. do.
- One aspect provides Micrococcus cohnii strains.
- the strain may be isolated.
- isolated means something that is artificially separated and available that does not exist in nature.
- the strain may be isolated from human skin.
- the strain may be isolated from human keratinocytes.
- the strain may be a strain isolated by a method of inoculating human keratinocytes into R2A (Reasoner's 2A) medium (Becton Dickinson, Cockeysville, MD) and culturing the cultured colony in pure isolation.
- the strain may be a strain belonging to the Micrococcus sp.
- the strain comprises a 16S rRNA having at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9% sequence identity with SEQ ID NO: 1 it may be
- the strain may contain the 16S rRNA of SEQ ID NO: 1.
- sequence identity refers to the degree of identity of amino acid residues or bases between sequences after aligning both sequences in a specific comparison region so as to match each other as much as possible. Sequence identity can be confirmed according to methods known in the art. The percent of sequence identity can be determined using a known sequence comparison program, examples of which include BLASTN (NCBI), CLC Main Workbench (CLC bio), MegAlignTM (DNASTAR Inc), and the like.
- the Micrococcus cohnii strain may be a strain deposited under accession number KCCM12996P.
- the strain deposited with the accession number may be Micrococcus conii MC-2 strain.
- the strain may have a skin condition improving effect.
- the skin condition improvement may be one or more of skin aging prevention, skin wrinkle improvement, skin inflammation improvement, atopy improvement, itching inhibition, and skin whitening.
- Another aspect provides a lysate of the strain, a culture medium, or an extract of the culture medium.
- lysate may be used interchangeably with “lysate” and may refer to a product obtained by disrupting the cell wall of a strain by chemical or physical force.
- the homogenate may include the homogenate itself, a concentrate or a lyophilized product thereof.
- the term "culture medium” may be used interchangeably with “culture supernatant”, “conditioned culture medium” or “conditioned medium”, and may supply nutrients so that Micrococcus conii strains can grow and survive in vitro. It may mean the entire medium including the strain obtained by culturing the strain in a medium for a certain period of time, its metabolites, extra nutrients, and the like.
- 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", and the culture solution is left still for a certain period of time to take only the liquid of the upper layer except for the part sunk in the lower layer, remove the cells through filtration, or centrifuge the culture solution to 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 isolated and selected from a sample or the strain isolated 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 to the lower layer, or it can be obtained by leaving it for a certain period of time and then removing the upper liquid because it sinks to the lower layer of the culture medium by gravity.
- the culture solution may include the culture solution itself obtained by culturing the strain, its concentrate or lyophilisate, or the culture supernatant obtained by removing the strain from the culture solution, its concentrate or lyophilisate.
- the culture medium and culture conditions for culturing the Micrococcus conii strain can be appropriately selected or modified by those skilled in the art.
- the culture medium obtained by culturing in a medium eg, R2A medium, RCM medium or TSA medium
- a medium eg, R2A medium, RCM medium or TSA medium
- culture broth extract refers to an extract obtained from the culture medium or a concentrate thereof, and may include an extract, a dilution 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.
- the strain, a lysate thereof, a culture medium, or an extract of the culture medium may be biologically pure.
- the strain may be obtained by pure culture.
- Another aspect provides the use of a Micrococcus conii strain, a lysate, culture, or extract of the strain. Specifically, it provides a composition comprising a Micrococcus conii strain, a lysate thereof, a culture medium, or an extract of the culture medium.
- the Micrococcus conii strain may be the Micrococcus conii strain according to the above aspect.
- the strain may be isolated from human skin.
- the strain has at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9% sequence identity with SEQ ID NO: 1 It may include 16S rRNA having
- the strain may be one containing the 16S rRNA of SEQ ID NO: 1.
- the strain may be a strain deposited with accession number KCCM12996P.
- the use of the strain may include improving skin conditions, improving skin beauty, and preventing, improving or treating skin diseases.
- the skin condition improvement or skin beauty improvement may be one or more of skin aging prevention, skin wrinkle improvement, skin inflammation improvement, atopy improvement, itching inhibition, and skin whitening.
- This strain increases COL1A1 expression; decrease MMP-1 expression; reduce IL-6 expression; It may be inhibiting melanin production.
- skin aging refers to both tangible and intangible changes that appear in the skin with age, such as thinning of the epidermis, decrease in the number of cells or blood vessels in the dermis, decrease in DNA damage repair ability, cell replacement Cycle reduction, wound healing ability, skin barrier function, epidermal moisture retention function, vitamin D production, physical damage defense function, chemical substance removal ability, immune response, sensory function, body temperature control, etc. says
- the skin aging may be 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 not only premature aging symptoms induced by external stimuli such as ultraviolet rays, pollution, cigarette smoke, chemicals, etc., but also a natural aging phenomenon that occurs as the proliferation of skin cells decreases with age. 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.
- improvement of inflammation may be used interchangeably with “inhibition of inflammation” and “anti-inflammation”, and may refer to any action in which NO production is suppressed by alleviating the immune response.
- skin disease may be a disease or skin inflammation due to skin barrier function impairment.
- prevention includes inhibiting the occurrence of a disease.
- treatment includes the inhibition, alleviation, or elimination of the development of a disease.
- the skin inflammatory disease may be any one selected from the group consisting of dermatitis, atopic dermatitis, pruritus (pruritus), eczematous skin disease, dry eczema, erythema, urticaria, psoriasis, drug rash, and acne.
- skin whitening may mean not only brightening the skin tone by inhibiting the synthesis of melanin pigment, but also improving skin hyperpigmentation such as spots or freckles caused by ultraviolet rays, hormones, or heredity.
- composition according to one aspect increases the expression of COL1A1 or reduces the expression of MMP-1 by including the Micrococcus conii strain, a lysate thereof, a culture medium or an extract of the culture medium, thereby exhibiting effects of preventing skin aging and improving skin wrinkles.
- composition according to one aspect reduces the expression of inflammatory cytokines (eg, IL-6) by including the Micrococcus conii strain, its lysate, culture medium or extract of the culture medium, thereby improving skin inflammation, or skin inflammatory disease, atopy. , It can prevent, improve or treat skin diseases such as itching.
- inflammatory cytokines eg, IL-6
- composition according to one aspect may exhibit a skin whitening effect by suppressing melanin production by including the Micrococcus conii strain, a lysate thereof, a culture medium, or an extract of the culture medium.
- the strain may exhibit a synergistic effect when used together with other strains having a skin condition improving effect, for example, a strain belonging to the genus Micrococcus.
- 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 the strain, a lysate thereof, a culture medium, or an extract of the culture medium.
- the term "included as an active ingredient” means that the strain of the present specification, its lysate, culture medium, or extract of the culture medium is added to the extent that the above-mentioned effects can be exhibited, and drug delivery and stabilization, etc. It means that it is formulated in various forms by adding various components as subcomponents for the purpose.
- the composition may be in a liquid or dry state.
- the composition may be in the form of a dry powder.
- a drying method for preparing the composition in a dry state may use a method commonly used in the art, and is not particularly limited.
- Non-limiting examples of the drying method include an air drying method, a natural drying method, a spray drying method, a freeze drying method, and the like. These methods may be used alone or at least two methods may be used together.
- the composition may contain an additive in an effective amount sufficient to reduce deterioration of the strain, its lysate, culture medium or extract of the culture medium.
- the additive may be, for example, a binder, but is not limited thereto.
- the composition may further include a cosmetically, pharmaceutically or food-acceptable carrier.
- the composition may be formulated with a carrier and provided as cosmetics, drugs, food additives, and the like.
- the composition may be a cosmetic composition.
- the cosmetic composition may further include ingredients commonly used in cosmetic compositions, functional additives, etc. in addition to the active ingredients disclosed herein, such as antioxidants, stabilizers, solubilizers, surfactants, dispersants, emulsifiers, and preservatives. , conventional adjuvants such as vitamins, pigments, flavors, and the like, and carriers.
- the cosmetic composition is not particularly limited to a specific formulation, and the formulation may be appropriately selected according to the purpose.
- the cosmetic composition may have, for example, a solubilizing formulation, an emulsifying formulation, or a dispersion formulation.
- the cosmetic composition may have, for example, a softening lotion, a nutrient lotion, a massage cream, a nutrient cream, an essence, a pack, a gel, an ampoule, or a skin-adhesive cosmetic formulation.
- 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.
- metal sequestering agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid
- drugs such as caffeine, tannin, bellapamil, licorice extract, glabridin, hot water extract of calin fruit, various herbal medicines, tocopherol acetate, glycylitnic acid, tranexamic acid and its derivatives or salts
- Vitamin C magnesium ascorbyl phosphate, ascorbic acid glucoside, arbutin, kojic acid
- Sugars such as glucose, fructose, and 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, mannitol, 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 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, liquids, tablets, capsules, dry syrups, and the like.
- Parenteral dosage forms may be injections, ointments, and the like.
- the composition may be a health functional food composition.
- it may be formulated in the form of a conventional health functional food known in the art.
- the health functional food composition may be used alone, or in combination with other foods or food ingredients, and may be appropriately used according to conventional methods.
- the mixing amount of the active ingredient may be appropriately determined depending on the purpose of use (prevention, health or therapeutic treatment).
- 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; and polysaccharides such as dextrins and cyclodextrins; It is a sugar alcohol, 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 food composition is also used in 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, alcohols, carbonated beverages carbonation agent, or a combination thereof.
- the 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, ameliorating, or treating a condition in a subject in need thereof comprising administering to a subject in need thereof an effective amount of a composition described above.
- the condition of the subject may be a condition related to skin or a condition related to skin inflammation.
- administering introducing
- implanting are used interchangeably herein, and according to one embodiment, a subject by a method or route that results in at least partial localization of a composition to a desired site. It can mean the placement of a composition according to one embodiment into.
- Administration may be administered by a method known in the art. Administration may 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. can 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 requiring skin improvement, for example, a subject requiring skin aging prevention, skin wrinkle improvement, skin inflammation improvement, atopy improvement, itching inhibition, or skin whitening effect.
- 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 two or more times within the range of clinically acceptable side effects, and administration can be performed at one or two or more sites, daily or at intervals of 2 to 5 days, for a total of
- 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 novel Micrococcus conii strain since it has effects of preventing skin aging, improving skin wrinkles, improving skin inflammation, improving atopy, inhibiting itching, and skin whitening, it can be applied in various ways for skin improvement, prevention, or treatment. can
- 1 is a result showing the relative expression level of COL1A1 in fibroblasts.
- 3 is a result showing the relative expression level of IL-6 in keratinocytes.
- Figure 4 is a result showing the melanin content (%) in mouse pigment cells.
- Micrococcus conii strains were isolated and identified from the skin of healthy women.
- samples obtained by washing the skin of healthy women with sterile distilled water that is, human epidermal keratinocytes
- R2A Reasoner's 2A
- Becton Dickinson Cockeysville, MD.
- 100 colonies formed after culturing in an incubator at 28° C. for 48 hours were cultured in pure isolation and re-cultured in an incubator at 28° C. for 48 hours.
- the cultured colony was subjected to 16S rRNA gene sequence identification. Specifically, PCR was performed using primers designed to amplify in response to bacteria only. 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. After the PCR reaction, the DNA sequences of the isolated and cultured species were determined using ABI-3730XL (ABI, USA).
- MC-2 microorganism Micrococcus cohnii strain
- COL1A1 Collagen, type I, alpha 1
- MMP-1 Mestrix metalloproteinase-1
- a human dermal fibroblast (Hs68) cell line (Human dermal fibroblast, Hs68) was dispensed in 3.5 ⁇ 10 5 in a 6-well plate, and then cultured for 24 hours in an incubator at 37° C. and 5% CO 2 conditions. Thereafter, the medium was removed, DPBS (Dulbecco's phosphate-buffered saline) was added, and 12 mJ/cm 2 of UVB was irradiated or not irradiated. Immediately after UVB irradiation, DPBS was removed and replaced with FBS-free medium, and then treated with 1% (w/w) of the culture medium of the MC-2 strain and further cultured for 24 hours. As a negative control group, UV treatment and strain culture solution non-treatment groups were used.
- cDNA was synthesized (C1000 Thermal Cycler, Bio-Rad, USA).
- cybergreen SYBR Green supermix, Applied Biosystems, USA
- a real-time PCR machine (Step One Plus, Applied Biosystems, USA) was used.
- Real-time polymerase chain reaction was performed at 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.
- Example 1 In order to analyze whether the culture solution of the MC-2 strain isolated in Example 1 has anti-inflammatory, anti-atopic and anti-itching activities, the expression of the inflammatory cytokine IL-6 was confirmed.
- human keratinocyte HaCaT cells were cultured in DMEM medium (Dulbecco's modified Eagle's Medium, Gibco 1210-0038) containing 10% fetal bovin serum. % CO 2 was performed in an incubator. The medium was exchanged every 3 to 4 days, and subculture was performed when the cells proliferated excessively. Thereafter, the cells were dispensed at 5 ⁇ 10 5 /well and washed with phosphate buffered saline solution (PBS) after 24 hours of culture. Cells were seeded into each well containing DMEM medium without FBS, and inflammation was induced in the keratinocyte cell line by adding 10 ⁇ g/ml of Poly I:C and 10 ng/ml of IL-4.
- DMEM medium Dulbecco's modified Eagle's Medium, Gibco 1210-003082
- PBS phosphate buffered saline solution
- the culture medium of the above strain was treated at a concentration of 1% (w/w), and further cultured for 4 hours.
- 1 ⁇ M of dexamethasone was used as a positive control group.
- the expression level of IL-6 was analyzed in the same manner as in Experimental Example 1, except that the IL-6 primer was used.
- the expression level of the gene was finally analyzed through correction for the ⁇ -actin gene, and the results are shown in FIG. 3 .
- MC-2 strain can be usefully used for the prevention, improvement, or treatment of skin inflammatory diseases, atopy, and itching.
- mouse pigment B16-F10 (Murine Melanoma) cells ATCC were suspended in 2 ml of DMEM medium (Dulbecco's modified Eagle's Medium, Gibco 1210-0038) containing 10% FBS to obtain 1 x 10 5 cells/ After inoculation into a 6-well plate, the cells were cultured for 24 hours in a 37°C, 5% CO 2 incubator until 40 to 50% of the cells adhered to the bottom of the well. Thereafter, ⁇ -MSH was treated at a concentration of 100 nM to induce melanin production. Next, 1% (w/w) and 10% (w/w) of the culture medium of the MC-2 strain were treated and additionally cultured for 3 days.
- DMEM medium Dulbecco's modified Eagle's Medium
- the MC-2 strain can be usefully used for skin whitening.
- the Micrococcus conii MC-2 strain increased COL1A1 expression and decreased MMP-1 expression; reduce IL-6 expression; It was confirmed that the melanin production was inhibited. Accordingly, it was found that the Micrococcus conii MC-2 strain had anti-aging effects, improved skin wrinkles, improved skin inflammation, improved atopy, suppressed skin itching, and skin whitening effects.
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Abstract
La présente invention concerne : une souche de Microccocus cohnii MC -2 ; un lysat, une solution de culture et une solution d'extrait de celui-ci ; une composition le comprenant ; et son utilisation pour améliorer l'état de la peau. Une nouvelle souche de Microccocus cohnii selon un mode de réalisation a pour effet de prévenir le vieillissement de la peau, la réduction des rides de la peau, le soulagement de l'inflammation de la peau, le soulagement de la dermatite atopique, l'inhibition du prurit et le blanchiment de la peau, et peut ainsi être utilisé de diverses manières pour l'amélioration, la prévention ou le traitement de la peau.
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WO2018073278A1 (fr) * | 2016-10-18 | 2018-04-26 | Universite Paris Descartes | Composition cosmétique pour prévenir et/ou traiter une odeur corporelle |
KR102273232B1 (ko) * | 2020-05-08 | 2021-07-06 | 코스맥스 주식회사 | 마이크로코커스 리래 균주 및 그의 피부 상태 개선 용도 |
KR102273233B1 (ko) * | 2020-05-08 | 2021-07-06 | 코스맥스 주식회사 | 마이크로코커스 루테우스 균주 및 그의 피부 상태 개선 용도 |
-
2021
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2018073278A1 (fr) * | 2016-10-18 | 2018-04-26 | Universite Paris Descartes | Composition cosmétique pour prévenir et/ou traiter une odeur corporelle |
KR102273232B1 (ko) * | 2020-05-08 | 2021-07-06 | 코스맥스 주식회사 | 마이크로코커스 리래 균주 및 그의 피부 상태 개선 용도 |
KR102273233B1 (ko) * | 2020-05-08 | 2021-07-06 | 코스맥스 주식회사 | 마이크로코커스 루테우스 균주 및 그의 피부 상태 개선 용도 |
Non-Patent Citations (2)
Title |
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EL-MOGHEITH SAHAR ABD, EL-GENDY AHMED OSAMA , SULTAN SERAGELDEEN , EL-NESR KHALID A. : "Exploring the Antimicrobial and Hepatoprotective Effects of Kefir; A Probiotic Fermented Milk", JOURNAL OF PURE AND APPLIED MICROBIOLOGY, ORIENTAL SCIENTIFIC PUBLISHING COMPANY, IN, vol. 11, no. 2, 30 June 2017 (2017-06-30), IN , pages 759 - 772, XP093034691, ISSN: 0973-7510, DOI: 10.22207/JPAM.11.2.15 * |
RIESER GERNOT, SCHERER SIEGFRIED, WENNING MAREIKE: "Micrococcus cohnii sp. nov., isolated from the air in a medical practice", INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, SOCIETY FOR GENERAL MICROBIOLOGY, GB, vol. 63, no. Pt_1, 1 January 2013 (2013-01-01), GB , pages 80 - 85, XP093034690, ISSN: 1466-5026, DOI: 10.1099/ijs.0.036434-0 * |
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