WO2024025173A1 - Composition cosmétique comprenant une souche de staphylococcus sp., une souche de streptococcus sp., une souche de cutibacterium sp., une souche d'enhydrobacter sp. et un mélange de fragments d'adn - Google Patents

Composition cosmétique comprenant une souche de staphylococcus sp., une souche de streptococcus sp., une souche de cutibacterium sp., une souche d'enhydrobacter sp. et un mélange de fragments d'adn Download PDF

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WO2024025173A1
WO2024025173A1 PCT/KR2023/009049 KR2023009049W WO2024025173A1 WO 2024025173 A1 WO2024025173 A1 WO 2024025173A1 KR 2023009049 W KR2023009049 W KR 2023009049W WO 2024025173 A1 WO2024025173 A1 WO 2024025173A1
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strain
culture medium
culture
lysate
extract
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PCT/KR2023/009049
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English (en)
Korean (ko)
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조형우
이동걸
강승현
백승걸
강소민
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코스맥스 주식회사
주식회사 파마리서치
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Publication of WO2024025173A1 publication Critical patent/WO2024025173A1/fr

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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
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/60—Sugars; Derivatives thereof
    • 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
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin

Definitions

  • Microorganisms living on human skin are known to maintain a symbiotic relationship with their human hosts and have various effects on skin aging by interacting with humans. These microorganisms and various information, including their genes, are called skin microbiome.
  • NGS next generation sequencing
  • Human skin which is the habitat of these microorganisms, comes in contact with the external environment and serves as a collection site for various microorganisms (e.g., fungi, bacteria, viruses, and small larvae).
  • various microorganisms e.g., fungi, bacteria, viruses, and small larvae.
  • the microorganisms prepare a habitat by adapting according to the selection of physical and chemical functions.
  • the epidermis forms the skin barrier and plays an important role in blocking the penetration of microorganisms and toxins and maintaining moisture.
  • the uppermost layer of the epidermis is composed of the stratum corneum.
  • the epidermis has a form called a ‘brick and mortar structure’. Skin tissue goes through a continuous self-recovery process, and scales that have completed the differentiation process constantly repeat the process of being shed from the skin tissue. It has been revealed that microorganisms influence these changes in the skin.
  • MMPs matrix metalloproteinases
  • photoaging is considered the biggest aging factor of the skin, and accordingly, many products to prevent photoaging are being released.
  • photoaging prevention technology using natural materials is limited in use and various problems arise when verifying efficacy. There are limitations in many cases.
  • One aspect is a Staphylococcus sp. strain, its lysate, culture, extract of the culture, or a mixture thereof; Streptococcus sp. strain, its lysate, culture, extract of the culture, or mixtures thereof; Cutibacterium sp. strain, its lysate, culture, extract of the culture, or mixtures thereof; Enhydrobacter sp. strain, its lysate, culture medium, extract of the culture medium, or mixtures thereof; and a composition for improving skin condition comprising a DNA fragment mixture as an active ingredient.
  • Another aspect provides a method of preventing, ameliorating, or treating a condition in a subject comprising treating or administering an effective amount of the composition described above to the subject in need thereof.
  • Another aspect includes the above-mentioned four strains, their lysate, culture medium, extract of the culture medium, or mixtures thereof for use in preparing a composition for improving skin condition; and the use of DNA fragment mixtures.
  • One aspect is a Staphylococcus sp. strain, its lysate, culture, extract of the culture, or a mixture thereof; Streptococcus sp. strain, its lysate, culture, extract of the culture, or mixtures thereof; Cutibacterium sp. strain, its lysate, culture, extract of the culture, or mixtures thereof; Enhydrobacter sp. strain, its lysate, culture medium, extract of the culture medium, or mixtures thereof; and a DNA fragment mixture as an active ingredient.
  • lysate may be used interchangeably with “lysate” and may refer to a product obtained by breaking the cell wall of a strain by chemical or physical force.
  • the lysate may include the lysate itself, its concentrate, or its freeze-dried product.
  • the term “culture” may be used interchangeably with “culture supernatant,” “conditioned culture,” or “conditioned medium,” and may be used interchangeably with the genus Staphylococcus, Streptococcus, Cutibacterium, and/or Inhydro. It may refer to the entire medium containing the strain, its metabolites, extra nutrients, etc. obtained by culturing the strain for a certain period of time in a medium that can supply nutrients so that the Bacter strain can grow and survive in vitro. Additionally, the culture medium may refer to a culture medium obtained by removing the bacterial cells from the bacterial culture medium obtained by culturing the strain.
  • the liquid from which the bacteria have been removed from the culture medium is also called “supernatant", and the supernatant is obtained by leaving the culture solution still for a certain period of time and taking only the upper liquid excluding the part that has settled in the lower layer, or by removing the bacteria through filtration, or by removing the culture liquid. It can be obtained by centrifuging to remove the sediment at the bottom and taking only the liquid at the top.
  • the "bacteria” refers to the strain itself of the present invention, and includes the strain itself separated and selected from a sample, etc., or a strain isolated from the culture medium by culturing the strain.
  • the bacterial cells can be obtained by centrifuging the culture medium and taking the part that has sunk to the lower layer. Alternatively, since they sink to the lower layer of the culture medium by gravity, they can be obtained by leaving them still for a certain period of time and then removing the upper liquid.
  • the culture medium includes strains of the genus Staphylococcus, Streptococcus, Cutibacterium, and/or Inhydrobacter in a medium (e.g., R2A, TSB, TSA, BHI or RCM medium) at an appropriate temperature (e.g. : It may be obtained by culturing for a certain period of time (e.g., 4 to 120 hours) at any temperature above 10°C or below 40°C.
  • a medium e.g., R2A, TSB, TSA, BHI or RCM medium
  • an appropriate temperature e.g. : It may be obtained by culturing for a certain period of time (e.g., 4 to 120 hours) at any temperature above 10°C or below 40°C.
  • the culture medium may include the culture medium itself, its concentrate, or freeze-dried product obtained by cultivating the strain, or the culture supernatant obtained by removing the strain from the culture medium, its concentrate, or freeze-dried product.
  • the culture supernatant of the strain may be obtained by removing the strain by centrifuging or filtering the strain culture fluid.
  • the concentrate may be obtained by concentrating the strain culture itself or the supernatant obtained after centrifuging or filtering the culture.
  • culture extract refers to an extract from the culture medium or its concentrate, and may include extracts, diluted or concentrated extracts, dried products obtained by drying the extracts, crude or purified products thereof, and fractions thereof. You can.
  • the strain, its lysate, culture medium, or extract of the culture medium may be biologically pure.
  • the strain may be obtained by pure culture.
  • a mixture of strains and DNA fragments of the genus Staphylococcus, Streptococcus, Cutibacterium, and Inhydrobacter increase the expression of genetic markers COL1A1 and elastin (ELN), which are reduced by ultraviolet rays, and increase the expression of genes related to skin moisturization. It was confirmed that the expression of factors (eg, HAS3) and skin barrier-related factors (eg, ABCA12) was increased. Therefore, the composition can be used to improve skin condition, improve skin beauty, prevent, improve or treat skin diseases, etc.
  • factors eg, HAS3
  • skin barrier-related factors eg, ABCA12
  • the improvement of skin condition or skin beauty may be preventing skin aging (anti-aging), improving skin wrinkles (anti-wrinkle), improving skin elasticity, moisturizing skin, or strengthening the skin barrier.
  • skin aging refers to both tangible and intangible changes that occur in the skin as one ages. Skin aging includes, for example, a thinning of the epidermis, a decrease in the number of cells or blood vessels in the dermis, a decrease in the ability to repair DNA damage, an increase in the cell turnover cycle, a decrease in the ability to heal wounds, a weakening of the skin barrier function, a decrease in the moisture retention ability of the epidermis, These include reduced sweat secretion, reduced sebum secretion, reduced vitamin D production, reduced ability to defend against physical damage, reduced ability to eliminate chemicals, reduced immune response, weakened sensory function, and reduced body temperature regulation.
  • the prevention of skin aging may mean improving skin aging caused by extrinsic or endogenous factors.
  • the exogenous factors refer to various external factors, such as ultraviolet rays (light).
  • the endogenous factors are also referred to as chronological factors and mainly refer to factors that occur due to the passage of time.
  • the skin aging is specifically not only a symptom of premature aging 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, etc.
  • wrinkles include stimulation caused by changes in internal and external factors that change the components that make up skin tissue, causing wrinkles.
  • the aging may be photoaging.
  • photoaging is a phenomenon caused by external environmental factors, the most representative factor being ultraviolet rays. Ultraviolet rays cause damage to biological components such as activation of proteolytic enzymes, chain cutting of matrix proteins, and abnormal cross-linking, and repetition of this mechanism causes skin aging that is evident in appearance.
  • wrinkle refers to a state in which the skin loses its elasticity and becomes loose, for example, the skin may be folded.
  • prevention or improvement of skin wrinkles may refer to any action that prevents or improves wrinkles by suppressing the expression of factors related to wrinkles, or increases the total amount of collagen.
  • skin moisturizing may refer to any action that maintains skin moisture or prevents moisture loss.
  • skin barrier strengthening may refer to any action that improves the function of the skin barrier, which is located on the outermost layer of the skin and prevents moisture and nutrition loss.
  • the “skin disease” may be a disease caused by damage to the skin barrier function, skin aging, skin wound, skin scar, or skin inflammation.
  • prevention includes suppressing the occurrence of a disease.
  • treatment includes inhibiting, alleviating, or eliminating the development of a disease.
  • improvement may refer to any action that results in at least reducing the severity of a parameter associated with the alleviation or treatment of a condition, such as symptoms.
  • the damage to the skin barrier function may mean any change that appears in the skin due to decreased or damaged skin barrier function. For example, it may include increased skin wrinkles, dryness, dermatitis, atopic dermatitis, allergic dermatitis, acne, etc.
  • the Staphylococcus sp. strains include, for example, Staphylococcus epidermidis , Staphylococcus hominis , Staphylococcus warneri , Staphylococcus caprae , It may be Staphylococcus capitis , etc.
  • the Staphylococcus genus strain may be Staphylococcus epidermidis (Accession Number: KCCM12559P).
  • the Staphylococcus genus strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof may be included in an amount of 5 to 20% by weight based on the total weight of the composition.
  • the Staphylococcus genus strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof is, for example, 5 to 20% by weight, 5 to 18% by weight, 5 to 16% by weight, 7 to 7% by weight, based on the total weight of the composition. It may be included at 20% by weight, 7 to 17% by weight, 8 to 15% by weight, or 10 to 13% by weight.
  • Streptococcus genus ( Streptococcus sp.) strains are, for example, Streptococcus thermophiles, Streptococcus mitis , Streptococcus infantis , It may be Streptococcus pneumoniae , etc.
  • the Streptococcus genus strain may be Streptococcus thermophiles (Accession number: KCCM12004P).
  • the Streptococcus genus strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof may be included in an amount of 5 to 30% by weight based on the total weight of the composition.
  • the Streptococcus genus strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof is contained in an amount of 5 to 30% by weight, 5 to 28% by weight, 5 to 25% by weight, 5 to 20% by weight, 8%, based on the total weight of the composition. It may be included in an amount of from 30% by weight, 8 to 24% by weight, 8 to 18% by weight, 10 to 27% by weight, 10 to 20% by weight, or 10 to 15% by weight.
  • the Cutibacterium sp. strain may be, for example, Cutibacterium acnes , Cutibacterium granulosum , etc.
  • the strain of the Cutibacterium genus may be Cutibacterium acnes (Accession Number: KCCM12680P).
  • the Cutibacterium genus strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof may be included in an amount of 50 to 90% by weight based on the total weight of the composition.
  • the Cutibacterium genus strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof is, for example, 50 to 90% by weight, 50 to 80% by weight, 50 to 70% by weight, 55 to 55% by weight, based on the weight of the species of the composition. It may be included at 90% by weight, 55 to 85% by weight, 55 to 75% by weight, 60 to 90% by weight, 60 to 80% by weight, or 70 to 75% by weight.
  • the Enhydrobacter sp. strain may be, for example, Enhydrobacter aerosaccus (Accession number: KCCM13073P).
  • the Inhydrobacter genus strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof may be included in an amount of 0.5 to 10% by weight based on the total weight of the composition.
  • the Inhydrobacter genus strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof is, for example, 0.5 to 10% by weight, 0.5 to 8% by weight, 0.5 to 5% by weight, 1 to 1% by weight, based on the total weight of the composition. It may be included at 7% by weight, 1 to 6% by weight, 2 to 9% by weight, or 2 to 5% by weight.
  • DNA fragment mixture is a form in which DNA corresponding to a biopolymer composed of phosphoric acid, four types of bases, and deoxyribose is extracted and exists in a mixture of DNA fragments having various molecular weights.
  • the DNA fragment mixture may be hydrolyzed.
  • the DNA fragment mixture may be, for example, polydeoxyribonucleotide (PDRN), polynucleotide, etc.
  • the polydeoxyribonucleotide (PDRN) is a mixture of short deoxyribonucleotides, a low molecular weight DNA complex made by fractionating DNA chains to a certain size. It's kind of like that.
  • the polynucleotide refers to a DNA or RNA strand of a certain length or more, which is a polymer of nucleotides in which nucleotide monomers are connected in a chain shape by covalent bonds.
  • the polynucleotide may have a relatively longer nucleic acid length or a higher molecular weight than a polydeoxyribonucleotide, but is not limited thereto.
  • the molecular weight of the DNA fragment mixture may be 50 to 10,000 kDa.
  • the molecular weight of the DNA fragment mixture may be, for example, 50 to 10,000 kDa, 50 to 5,000 kDa, 50 to 3,000 kDa, 50 to 1,500 kDa, 1,000 to 10,000 kDa, 1,000 to 5,000 kDa, or 5,000 to 10,000 kDa.
  • the DNA fragment mixture may be a mixture of DNA fragments obtained by extraction from testes or semen of a fish, and the fish may be a salmonid, specifically salmon or trout.
  • the DNA fragment mixture may be included in an amount of 0.01 to 1% by weight based on the total weight of the composition.
  • the DNA fragment mixture may be included, for example, at 0.01 to 1% by weight, 0.01 to 0.7% by weight, 0.03 to 0.7% by weight, 0.1 to 0.8% by weight, or 0.3 to 0.6% by weight, based on the total weight of the composition. At this time, if the content of the DNA fragment mixture is less than or exceeds the above range, there is a problem that the skin condition improvement effect cannot be sufficiently exhibited.
  • the composition may have synergistic activity in improving skin condition by containing strains of the genus Staphylococcus, Streptococcus, Cutibacterium, and Inhydrobacter and a mixture of DNA fragments in a specific weight ratio.
  • the mixture of strains and DNA fragments of the Staphylococcus genus, Streptococcus genus, Cutibacterium genus, and Inhydrobacter genus is, for example, 1:1 to 2:5 to 7:0.1 to 0.5:0.01 to 0.06, 1:1 It may be mixed at a weight ratio of 1.5:5 to 6.5:0.1 to 0.3:0.01 to 0.05, or 1:1 to 1.3:5.5 to 7:0.2 to 0.5:0.03 to 0.05.
  • the composition according to one aspect includes a Staphylococcus sp. strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof; Streptococcus sp. strain, its lysate, culture, extract of the culture, or mixtures thereof; Cutibacterium sp. strain, its lysate, culture, extract of the culture, or mixtures thereof; Enhydrobacter sp. strain, its lysate, culture medium, extract of the culture medium, or mixtures thereof; and a DNA fragment mixture in a specific content and weight ratio can exert a synergistic effect in improving skin condition.
  • the term "included as an active ingredient” refers to the strain of the present specification, its lysate, culture medium, or extract of the culture medium to the extent that it can exhibit the above-mentioned effects; and/or DNA fragment mixture is added, and includes formulation in various forms by adding various ingredients as sub-ingredients for drug delivery and stabilization.
  • the composition may be a cosmetic composition.
  • the cosmetic composition may further include ingredients commonly used in cosmetic compositions, functional additives, etc., such as antioxidants, stabilizers, solubilizers, surfactants, dispersants, emulsifiers, and preservatives. , conventional auxiliaries such as vitamins, pigments, fragrances, etc., and carriers.
  • the cosmetic composition is not particularly limited to a specific formulation, and the formulation may be appropriately selected depending on the purpose.
  • the cosmetic composition may have, for example, a solubilized formulation, an emulsified formulation, or a dispersed formulation.
  • the cosmetic composition includes, for example, lotion, cream, essence, cleansing foam, cleansing water, pack, ampoule, body lotion, body oil, body gel, shampoo, rinse, hair conditioner, hair gel, foundation, lipstick, mascara, and makeup. It may have a base or skin-adhesive cosmetic formulation.
  • the skin-adhesive cosmetic formulation may be, for example, a mask pack, but is not limited thereto.
  • the composition may be a composition for external application to the 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 skin external preparations include ingredients commonly used in external skin preparations such as cosmetics and medicines, such as aqueous ingredients, oil-based ingredients, powder ingredients, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, and fragrances. , colorants, various skin nutrients, metal ion sequestrants, sugars, or combinations thereof may be appropriately mixed as needed.
  • 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, and may be used appropriately according to conventional methods.
  • the mixing amount of the active ingredient can 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 materials.
  • beverage compositions may contain various flavoring agents or natural carbohydrates as additional ingredients like ordinary beverages.
  • the natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; and polysaccharides such as dextrins and cyclodextrins; Sugar alcohols such as xylitol, sorbitol, and erythritol.
  • a sweetener natural sweeteners such as thaumatin and stevia extract or synthetic sweeteners such as saccharin and aspartame can be used.
  • the health food composition also contains 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. It may contain the carbonating agent used, or a combination thereof.
  • the health functional food composition may also contain pulp for the production of natural fruit juice, fruit juice beverage, vegetable beverage, or a combination thereof.
  • Another aspect provides a method of preventing, ameliorating, or treating a condition in a subject comprising treating or administering an effective amount of the composition described above to the subject in need thereof.
  • the condition of the subject may be a condition related to skin-related conditions, and specifically, may be a disease caused by damage to the skin barrier function or skin aging.
  • administering As used herein, the terms “administering,” “introducing,” and “implanting” are used interchangeably and refer to an individual by a method or route that results in at least partial localization of the composition according to one embodiment to the desired site. It may refer to the arrangement of the composition according to one embodiment into the inner space.
  • Administration may be by methods known in the art.
  • the administration method such as administration route and administration frequency, can be appropriately selected by a person skilled in the art.
  • Administration may be administered directly to the subject by any means, such as, for example, intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal or subcutaneous administration. You can.
  • the administration may be administered systemically or locally.
  • the administration may include application to the skin.
  • the subject may be a mammal, such as a human, cow, horse, pig, dog, sheep, goat, or cat.
  • the subject may be an entity that requires improvement of skin condition, for example, an entity that requires anti-aging skin, improvement of skin wrinkles, enhancement of skin elasticity, skin moisturization, strengthening of skin barrier, and inhibition of skin inflammation.
  • the administration of the composition according to one embodiment 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 mg per day.
  • the dosage may be prescribed in various ways depending on factors such as formulation method, administration method, patient's age, weight, gender, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity, and those skilled in the art will Taking these factors into consideration, the dosage can be adjusted appropriately.
  • the frequency of administration can be once a day or two or more times within the range of clinically acceptable side effects.
  • the administration site it can be administered at one or two or more locations, and the total number of administration days can be from 1 to 30 days for one treatment, either daily or at intervals of 2 to 5 days. If necessary, the same treatment can be repeated after an appropriate period.
  • the dosage per kg is the same as for humans, or the above dosage is converted into, for example, the volume ratio (e.g., average value) of the organs (heart, etc.) between the target animal and human.
  • One dose can be administered.
  • Another aspect includes the above-mentioned four strains, their lysate, culture medium, extract of the culture medium, or mixtures thereof for use in preparing a composition for improving skin condition; and uses of DNA fragment mixtures.
  • Redundant content is omitted in consideration of the complexity of the present specification, and terms not otherwise defined in the present specification have meanings commonly used in the technical field to which the present invention pertains.
  • composition according to one aspect increases the expression of collagen, elastin, HAS3, and ABCA12, and can be usefully used to improve skin conditions such as skin aging, skin moisturizing, skin barrier strengthening, skin wrinkles, or skin elasticity.
  • Figure 1 is a result showing the effect of a composition according to one aspect on COL1A1 expression.
  • Figure 2 is a result showing the effect of a composition according to one aspect on ELN expression.
  • Figure 3 is a result showing the effect of a composition according to one aspect on HAS3 expression.
  • Figure 4 is a result showing the effect of a composition according to one aspect on ABCA12 expression.
  • Staphylococcus epidermidis strain culture medium was prepared. Specifically, Staphylococcus epidermidis strain (Accession number: KCCM12559P) was received from the Korean Culture Center of Microorganisms (KCCM) and grown at 150 rpm/min in R2A (Reasoner's 2A) medium. It was suspended and cultured for enrichment. The temperature was maintained at about 30°C during the culturing period, and after culturing for 72 hours, the culture medium was filtered using a 0.45 ⁇ m filter.
  • Staphylococcus epidermidis strain accesion number: KCCM12559P
  • R2A Reasoner's 2A
  • Streptococcus thermophiles strain (Accession number: KCCM12004P) and TSB (Tryptic Soy Broth) medium were used, except that Streptococcus thermophiles strain culture medium was prepared in the same manner as in Preparation Example 1 . did.
  • Cutibacterium acnes strain (Accession number: KCCM12680P) was prepared in the same manner as in Preparation Example 1 except that it was stationary cultured using RCM (Reinforced Clostridial Medium) medium. Culture medium was prepared.
  • An Enhydrobacter aerosaccus strain culture medium was prepared in the same manner as in Preparation Example 3, except that the Enhydrobacter aerosaccus strain (accession number: KCCM13073P) was used.
  • Example 1 Preparation of a composition comprising strain culture medium and DNA fragment mixture
  • a composition containing the ingredients and contents shown in Table 1 below was prepared. Specifically, a composition was prepared by mixing the culture media of Preparation Examples 1 to 4 with hydrolyzed DNA (manufactured by Pharma Research Co., Ltd.) with a molecular weight of 50 to 1,500 kDa.
  • the hydrolyzed DNA is a mixture of DNA fragments in a white or off-white powder. Unless otherwise stated herein, content is in weight percent.
  • composition was prepared in the same manner as Example 1, except that it included the ingredients and contents shown in Table 2 below.
  • Comparative Example 1 has the same mixing ratio of the strain culture medium as Example 1, but differs in that it contains sterilized distilled water instead of hydrolyzed DNA.
  • composition was prepared in the same manner as Example 1, except that it included the ingredients and contents shown in Table 3 below.
  • Comparative Example 2 differs from Example 1 in the mixing ratio of the strain culture medium.
  • the skin aging, wrinkle improvement, and skin elasticity enhancing activities of the composition according to one aspect were confirmed.
  • Human dermal fibroblast (Hs68) cell line was distributed at 3.5x10 5 in a 6-well plate and cultured in an incubator at 37°C and 5% CO 2 for 24 hours. Afterwards, the medium was removed, DPBS (Dulbecco's phosphate-buffered saline) was added, and 20 mJ/cm 2 UVB was irradiated or not. Immediately after UVB irradiation, DPBS was removed and replaced with medium without FBS fetal bovin serum, and then treated with the composition prepared in the above Examples and Comparative Examples and further cultured for 24 hours. As a negative control group, a group untreated with ultraviolet rays and strain culture was used.
  • DPBS Dens phosphate-buffered saline
  • cDNA was synthesized (C1000 Thermal Cycler, Bio-Rad, USA). Using the synthesized cDNA as a template, Cybergreen (SYBR Green supermix, Applied Biosystems, USA) and primers (primers for COL1A1 or primers for ELN) were added to a real-time PCR machine (Step One Plus, AppliedBiosystems, USA). Real-time polymerase chain reaction was performed. The expression level of the gene was finally analyzed through correction for the ⁇ -actin gene.
  • Figure 1 is a result showing the effect of a composition according to one aspect on COL1A1 expression.
  • Figure 2 is a result showing the effect of a composition according to one aspect on ELN expression.
  • composition according to one aspect increases the expression of COL1A1 and elastin, which are reduced by ultraviolet rays, by including four types of strain cultures and a mixture of DNA fragments as active ingredients, thereby preventing skin aging (e.g. photoaging) (anti-aging) ), is effective in improving wrinkles (anti-wrinkle) and improving skin elasticity.
  • skin aging e.g. photoaging
  • anti-aging anti-aging
  • the skin moisturizing activity of the composition according to one aspect was confirmed.
  • HaCaT cells a human keratinocyte cell line, were cultured in DMEM medium (Dulbecco's modified Eagle's Medium, Gibco 1210-0038) containing 10% fetal bovine serum, and all cultures were performed at 37°C and 5% CO 2 It was performed in an incubator.
  • the cultured cell lines were treated with the compositions prepared in the Examples and Comparative Examples and further cultured for 24 hours.
  • HAS3 gene expression level was analyzed using the same method as Experimental Example 1-2, except that primers for HAS3 (hyaluronic acid synthase) were used.
  • Figure 3 is a result showing the effect of a composition according to one aspect on HAS3 expression.
  • composition according to one aspect significantly increases the expression of HAS3 by including four types of strain cultures and a mixture of DNA fragments as active ingredients, and can be usefully used to improve skin moisturization.
  • the skin barrier function improvement activity of the composition according to one aspect was confirmed.
  • HaCaT cells a human keratinocyte cell line, were treated with polyIC (polyinosinic:polycytidylic acid; Poly I:C), a synthetic ribonucleic acid that promotes interferon production, and then cultured for 24 hours to stimulate the HaCaT cell line. Thereafter, cells aged by stimulation were treated with 1% (w/w) of the composition prepared in the above Examples and Comparative Examples and further cultured for 24 hours.
  • polyIC polyinosinic:polycytidylic acid
  • Poly I:C polyinosinic:polycytidylic acid
  • a synthetic ribonucleic acid that promotes interferon production
  • ABCA12 ATP-binding cassette sub-family A member 12
  • Figure 4 is a result showing the effect of a composition according to one aspect on ABCA12 expression.
  • composition according to one aspect significantly increases the expression of ABCA12 by including four types of strain cultures and a mixture of DNA fragments as active ingredients, and can be usefully used to improve the skin barrier.
  • the composition according to one aspect includes a mixture of four types of strain cultures and DNA fragments, thereby (i) increasing the expression of COL1A1 and ELN reduced by ultraviolet rays, (ii) increasing the expression of HAS3 effect, (iii) has the effect of increasing the expression of ABCA12.
  • the effects of (i) to (iii) were significantly better than those of Comparative Example 1, which did not contain a mixture of DNA fragments, or Comparative Example 2, which had a different mixing ratio of the strain culture medium.

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Abstract

La présente invention concerne une composition cosmétique comprenant une souche de Staphylococcus sp., une souche de Streptococcus sp., une souche de Cutibacterium sp., un bouillon de culture d'une souche d'Enhydrobacter sp. et un mélange de fragments d'ADN. La composition peut être efficacement utilisée dans la prévention, l'amélioration ou le traitement d'états cutanés.
PCT/KR2023/009049 2022-07-27 2023-06-28 Composition cosmétique comprenant une souche de staphylococcus sp., une souche de streptococcus sp., une souche de cutibacterium sp., une souche d'enhydrobacter sp. et un mélange de fragments d'adn WO2024025173A1 (fr)

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KR1020220093390A KR102569532B1 (ko) 2022-07-27 2022-07-27 스타필로코커스 속, 스트렙토코커스 속, 쿠티박테리움 속, 인하이드로박터 균주 및 dna 단편 혼합물을 포함하는 화장료 조성물

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KR102273950B1 (ko) * 2020-05-08 2021-07-08 코스맥스 주식회사 큐티박테리움 아크네스 아종 아크네스 균주 및 그의 피부 상태 개선 용도
KR102415344B1 (ko) * 2021-05-04 2022-06-30 주식회사 파마리서치 피부투과도를 높인 dna 단편 혼합물을 함유하는 피부주름개선 조성물

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