WO2023282508A1 - 효모유래 세포외소포 및 용해물을 함유하는 화장료 조성물 - Google Patents

효모유래 세포외소포 및 용해물을 함유하는 화장료 조성물 Download PDF

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WO2023282508A1
WO2023282508A1 PCT/KR2022/008942 KR2022008942W WO2023282508A1 WO 2023282508 A1 WO2023282508 A1 WO 2023282508A1 KR 2022008942 W KR2022008942 W KR 2022008942W WO 2023282508 A1 WO2023282508 A1 WO 2023282508A1
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Prior art keywords
cosmetic composition
skin
extracellular vesicles
active ingredient
extracellular
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PCT/KR2022/008942
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English (en)
French (fr)
Korean (ko)
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조일지
황순석
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주식회사래디안
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Publication of WO2023282508A1 publication Critical patent/WO2023282508A1/ko

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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
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9728—Fungi, e.g. yeasts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
    • A61K8/11—Encapsulated compositions
    • 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
    • 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
    • A61Q19/08—Anti-ageing preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
    • A61K2800/82—Preparation or application process involves sonication or ultrasonication

Definitions

  • the present invention relates to a cosmetic composition
  • a cosmetic composition comprising an active ingredient isolated from at least one of extracellular vesicles derived from Saccharomyces cerevisiae and lysates thereof.
  • Prostaglandin E2 Prostaglandin E2
  • COX-2 an enzyme that produces inflammatory cytokines
  • MMP muscle growth factor
  • a lot of free radicals are generated, and the antioxidant defense system of the skin is damaged by these free radicals, which accelerates skin aging by increasing wrinkles and relaxing the skin. Therefore, in order to reduce skin wrinkles and maintain elasticity, it is necessary to protect the skin by inhibiting generation of reactive oxygen species and free radicals, inhibiting inflammatory reactions, and promoting skin regeneration from wounds.
  • Substances known to be effective in improving skin wrinkles include adenosine and retinoic acid, but adenosine has little clinical efficacy, retinoic acid cannot be used for women of childbearing age, and has side effects such as erythema. Accordingly, functional cosmetics for skin anti-aging using active ingredients of plants or natural products have recently been attracting attention. However, functional cosmetics containing plant extracts as active ingredients have problems in that a foreign body sensation may occur in the process of evaporation of the solution after application to the skin, and the duration of the effect is short.
  • Extracellular vesicles are classified into exosomes, microvesicles, and outer membrane vesicles according to their origin, size, and production process, but there is not much difference in appearance, and they are secreted outside the cell. It has a common feature of being a vesicle, so it is now collectively referred to as “extracellular vesicle”.
  • Exosomes are vesicles with a size of tens to hundreds of nanometers composed of a double-phospholipid membrane identical to the structure of a cell membrane, and contain proteins, nucleic acids (mRNA, miRNA, etc.) called exosome cargo inside. It is known that the exosome cargo contains a wide range of signaling factors, and these signaling factors are specific to cell types and differently regulated depending on the environment of the secreting cell.
  • Exosomes are intercellular signaling mediators secreted by cells, and various cell signals transmitted through them regulate cell behavior including activation, growth, migration, differentiation, dedifferentiation, apoptosis, and necrosis of target cells. It is known. Exosomes contain specific genetic materials and bioactive factors depending on the nature and state of the derived cell. In the case of proliferating stem cell-derived exosomes, cell behaviors such as cell migration, proliferation, and differentiation are regulated, and characteristics of stem cells related to tissue regeneration are reflected (Nature Review Immunology 2002 (2) 569-579). Due to these characteristics, studies have been conducted to utilize extracellular vesicles for purposes such as diagnosis/treatment (Patent Document 2).
  • Exosomes are known to be released not only from stem cells and cancer cells, but also from cells of various organisms such as plants, bacteria, fungi, and algae.
  • exosomes can be isolated from yeast cultures or fermented products in addition to mesenchymal stem cell cultures and fibroblast cultures.
  • fermented cosmetics are recently gaining popularity, and the types of yeast used in cosmetics are also increasing.
  • yeast used as raw materials for cosmetics There are about hundreds of types of yeast used as raw materials for cosmetics, and among fermented cosmetic raw materials, the best consumer response is Galactomyces Ferment Filtrate.
  • Galactomyces is a yeast found in the process of researching various yeasts, noting that the skin of the hands of craftsmen who make rice wine is elastic and clean compared to their age.
  • Galactomyces fermentation filtrate is rich in vitamins, minerals, amino acids, organic acids, enzymes, and yeast peptides.
  • Galactomyces fermentation filtrate may cause side effects such as skin trouble, reddening of the skin, or dermatitis in the case of people with skin sensitive to fermented ingredients (Patent Document 1).
  • Patent Document 1 Korean Patent Publication No. 10-2020-0028290
  • Patent Document 2 Korean Patent Publication No. 10-2019-0050286
  • the present invention is to provide a cosmetic composition for anti-aging, skin regeneration, skin elasticity improvement or skin wrinkle improvement containing exosomes derived from Saccharomyces cerevisiae and a lysate thereof as an active ingredient.
  • the culture solution contains various components such as components necessary for proliferation and metabolites emitted by cells, it contains both useful components and components harmful to the skin, To provide a cosmetic composition that produces excellent effects by separating only useful components.
  • the cosmetic composition according to the present invention preferably contains an active ingredient isolated from at least one of extracellular vesicles and lysates thereof derived from Saccharomyces cerevisiae.
  • yeast Saccharomyces cerevisiae RD-03 RI SC(C) (accession number: KCTC SD1359).
  • the cosmetic composition is for anti-aging, skin regeneration, skin elasticity improvement or skin wrinkle improvement.
  • the extracellular vesicles or lysates thereof are separated by ultracentrifugation or ultrafiltration.
  • the extracellular vesicles or a lysate thereof may be separated by an ultrafiltration membrane.
  • the size of the extracellular vesicles is 30 to 200 nm.
  • the cosmetic composition further comprises at least one additive selected from butylene glycol, hexanediol, ethylhexylglycerin and phenoxyethanol.
  • the cosmetic composition is a formulation selected from the group consisting of lotion, nutrient cream, massage cream, essence, pack, gel, lotion, ointment, patch, and spray.
  • the extracellular vesicles as a delivery vehicle for delivering the active ingredient of the extracellular vesicles or their lysate into the skin or into cells or extracellular vesicles.
  • the active ingredient is naturally encapsulated by injecting the active ingredient into extracellular vesicles during yeast culture using the extracellular vesicle delivery system, or ultrasonic waves after the active ingredient is introduced into extracellular vesicles during yeast culture
  • a method of artificially encapsulating in extracellular vesicles through treatment is preferred.
  • a cosmetic method for improving the condition of mammalian skin, except for treatment, using a cosmetic composition containing active ingredients isolated from at least one of extracellular vesicles derived from Saccharomyces cerevisiae and lysates thereof of the present invention can be used for
  • the cosmetic method includes (a) directly applying the cosmetic composition to the skin of a mammal, (b) contacting or attaching a patch, mask pack or mask sheet onto which the cosmetic composition is applied or deposited to the skin of a mammal. It is preferable to proceed sequentially through (a) and (b).
  • step (c) it is preferable to further include the step of removing the patch, mask pack or mask sheet from the skin of the mammal after step (b) and applying the cosmetic composition to the skin of the mammal.
  • the present invention is to provide a cosmetic composition for anti-aging, skin regeneration, skin elasticity improvement or skin wrinkle improvement containing only substantially useful ingredients from extracellular vesicles derived from Saccharomyces cerevisiae and lysates thereof as active ingredients. .
  • Example 1 shows a transmission electron microscope image of Example 2 according to the present invention.
  • the cosmetic composition according to the present invention preferably contains an active ingredient isolated from at least one of extracellular vesicles and lysates thereof derived from Saccharomyces cerevisiae.
  • a step when a step is said to be located “on” or “before” another step, this is not only the case where a step is in a direct time-series relationship with another step, but also the mixing step after each step As such, the order of the two steps may include the same rights as in the case of an indirect time-series relationship in which the time-series order may change.
  • the present invention provides a cosmetic composition
  • a cosmetic composition comprising an active ingredient isolated from at least one of extracellular vesicles and lysates thereof derived from Saccharomyces cerevisiae.
  • the cosmetic composition is for anti-aging, skin regeneration, skin elasticity improvement or skin wrinkle improvement.
  • yeast-derived extracellular vesicles and their lysates were obtained from yeast culture broth by two different separation methods. This was treated with human fibroblasts to confirm the culture conditions and extracellular vesicle separation method having the best collagen synthesis ability.
  • the ultracentrifugation method is mainly performed, but considering mass production, it is more preferable to use the ultrafiltration membrane used in the present invention, and if the process is established by setting the cutoff molecular weight of the ultrafiltration membrane in stages Subdivided commercialization, such as obtaining extracellular vesicles by particle size, will be possible.
  • the extracellular vesicles isolated in the present invention correspond to 30 to 200 nm, and yeast-derived extracellular vesicles can be regarded as naturally-derived nanostructures. Yeast-derived extracellular vesicles themselves can be used as products after confirming that they have the effect of preventing and improving skin wrinkles.
  • nano-sized structures are characterized by being easily combined with physiologically active substances that function to remove wrinkles or prevent aging.
  • Nanocosmetics are being developed by applying these nanostructures to cosmetics as a carrier, and are easily absorbed into the skin because their size is much smaller than the gap between skin cells. That is, the nanostructure can serve as a carrier that easily delivers cosmetic ingredients into the skin, into cells, or into extracellular vesicles.
  • nanostructures As a carrier and putting active ingredients into yeast-derived extracellular vesicles, it will be possible to derive more products.
  • a method of naturally encapsulating active ingredients by adding active ingredients through a transporter during yeast culture or a method of artificially encapsulating active ingredients by inducing structural changes in extracellular vesicles instantaneously through sonication can be used.
  • Yeast Sacharomyces cerevisiae RD-03 RI-SC(C) (KCTC SD1359) was inoculated into YM Broth and stirred for 8, 16, 24, 48 hours at 27°C and 37°C, followed by shaking incubation at 4°C , 4000 rpm, centrifuged under the conditions of 20 min, and the supernatant was filtered with 0.22 ⁇ m PES to prepare Comparative Examples 1 to 8.
  • Comparative Examples 9 to 16 were prepared by resuspending in a phosphate buffer.
  • Comparative Examples 17 to 24 were prepared by resuspending in distilled water and treating with sonication three times at 5-minute intervals.
  • Example 1 Yeast-derived extracellular vesicle solution using ultrafiltration
  • Yeast Sacharomyces cerevisiae RD-03 RI-SC(C) (KCTC SD1359) was inoculated into YM Broth, stirred and cultured at 27 ° C for 48 hours, centrifuged at 4 ° C, 4000 rpm, 20 min, and the upper layer The solution was filtered through 0.22 ⁇ m PES to prepare a yeast culture filtrate. The filtrate was removed by filtering with a 40 nm ultrafilter.
  • Example 1 was prepared by recovering EVs filtered through a 40 nm ultrafilter using 1/10 the amount of distilled water compared to the yeast culture filtrate and filtering through 0.22 ⁇ m PES.
  • Example 2 Yeast-derived extracellular vesicle lysate using ultrafiltration
  • Example 2 was prepared by sonicating the extracellular vesicle solution obtained in Example 1 three times at intervals of 5 minutes. Transmission electron microscopy images of these extracellular vesicle lysates are shown in FIG. 1 .
  • BCA assay was performed using samples of Comparative Examples 1 to 24.
  • Comparative Examples 1 to 8 in the form of yeast culture filtrate was the highest, and it was confirmed that the effect of the medium component was observed as the protein content tended to decrease as the culture time increased.
  • Comparative Examples 9 to 24 the protein content was less than 100 ⁇ g/ml, and it was confirmed that most proteins including medium components were removed during the separation of extracellular vesicles.
  • Comparative Example 12 and Comparative Example 20 had a high collagen synthesis ability, it was confirmed that Example 2 was higher.
  • Ergosterol also called provitamin D
  • provitamin D is one of the components contained in fungi such as yeast and mushrooms, and can be seen as a yeast EV membrane marker.
  • the presence or absence of ergosterol was confirmed using a thin layer chromatograph method.
  • Example 2 was lyophilized to remove the solvent, suspended in methanol, centrifuged, and 100 ⁇ l of the supernatant was added dropwise.
  • the Ergosterol standard was dissolved in methanol and dripped, developed in the developing solution, and it was confirmed whether a spot having the same Rf value as the standard was formed when irradiated with UV 254 nm.
  • Example 2 confirmed the presence of ergosterol by forming a spot having the same Rf value as that of the standard ergosterol product.
  • Example 2 was lyophilized, suspended in distilled water, loaded on a gel, and the protein pattern was analyzed using two protein markers of 10 to 180 kDa and 40 to 300 kDa.
  • Example 2 was confirmed to be composed of proteins of 60 to 75 kDa and 100 to 180 kDa.
  • the average particle size of Examples 1 and 2 was confirmed to be 150 ⁇ 220 nm, and the particle concentration was confirmed to be about 1 x 10 8 to 1 x 10 10 particles / ml.
  • the cosmetic composition may further include at least one additive selected from butylene glycol, hexanediol, ethylhexylglycerin, and phenoxyethanol. These additives are necessary for the formation of subsequent formulations, but are not limited thereto.
  • the cosmetic composition may be a formulation selected from the group consisting of softening lotion, nutrient lotion, nutrient cream, massage cream, essence, pack, gel, lotion, ointment, patch, and spray, but is not limited thereto.
  • a cosmetic composition containing an active ingredient isolated from at least one of extracellular vesicles derived from Saccharomyces cerevisiae and lysates thereof of the present invention improving the condition of mammalian skin except for treatment It can be used in cosmetic methods.
  • the cosmetic method includes (a) directly applying the cosmetic composition to the skin of a mammal, (b) contacting or attaching a patch, mask pack or mask sheet onto which the cosmetic composition is applied or deposited to the skin of a mammal. It is preferable to proceed sequentially through (a) and (b).
  • step (c) it is preferable to further include the step of removing the patch, mask pack or mask sheet from the skin of the mammal after step (b) and applying the cosmetic composition to the skin of the mammal.
  • the present invention relates to a cosmetic composition
  • a cosmetic composition comprising an active ingredient isolated from at least one of extracellular vesicles derived from Saccharomyces cerevisiae and lysates thereof.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
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PCT/KR2022/008942 2021-07-08 2022-06-23 효모유래 세포외소포 및 용해물을 함유하는 화장료 조성물 WO2023282508A1 (ko)

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KR1020210089480A KR20230008997A (ko) 2021-07-08 2021-07-08 효모유래 세포외소포 및 용해물을 함유하는 화장료 조성물
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CN116042422B (zh) * 2023-02-07 2024-03-26 广东丸美生物技术股份有限公司 一种酵母来源外泌体及其制备方法和应用

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