WO2020246766A1 - Composition pour prévenir ou atténuer le vieillissement cutané, contenant un extrait de rhodiola sachalinensis fermenté à l'aide de bovista plumbea - Google Patents

Composition pour prévenir ou atténuer le vieillissement cutané, contenant un extrait de rhodiola sachalinensis fermenté à l'aide de bovista plumbea Download PDF

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WO2020246766A1
WO2020246766A1 PCT/KR2020/007128 KR2020007128W WO2020246766A1 WO 2020246766 A1 WO2020246766 A1 WO 2020246766A1 KR 2020007128 W KR2020007128 W KR 2020007128W WO 2020246766 A1 WO2020246766 A1 WO 2020246766A1
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composition
extract
honggyeongcheon
fermented
skin
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PCT/KR2020/007128
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Korean (ko)
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권미진
조영호
윤영민
이현민
경기열
서태석
손영수
김관수
이주운
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(주)피에스에이
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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
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/41—Crassulaceae (Stonecrop family)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0014—Skin, i.e. galenical aspects of topical compositions
    • 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
    • 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
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/10—Preparation or pretreatment of starting material
    • A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
    • 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
    • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/52—Stabilizers
    • A61K2800/522—Antioxidants; Radical scavengers
    • 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
    • A61K2800/74—Biological properties of particular ingredients
    • A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
    • A61K2800/782—Enzyme inhibitors; Enzyme antagonists
    • 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
    • A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine

Definitions

  • the present invention relates to a composition for preventing or improving skin aging containing fermented honggyeongcheon extract, and more specifically, fermenting honggyeongcheon extract fermented liquid obtained after fermenting honggyeongcheon ( Rhodiola Sachalinensis ) extract using glutinous rice cake mushroom ( Bovista plumbea ) as an active ingredient It relates to a composition for preventing or improving skin aging containing as.
  • a typical external aging factor is ultraviolet rays, and the most prominent external aging phenomena are skin pigmentation and wrinkle formation (HW Daniell, Ann Intern Med, 1971; 75(6):873-880, GL Grove et al., J Am Acad Dermatol, 1989; 21(3):631-637, CE Griffiths et al., Arch Dermatol, 1992; 128: 347-351).
  • Free radicals are known to cause destruction of connective tissue formation such as skin collagen, inhibition of cell membrane function, promotion of DNA mutation, modification of protein action, energy transfer between cells, and modification of molecules related to metabolism (RM Lavker and AM Kligman, J Invest Dermatol, 1988; 90: 325-330).
  • the fact that free radicals are involved in aging means that antioxidants or various chemical scavengers that inactivate free radicals can be used to delay the aging process.
  • the skin is the primary barrier of the human body and protects the internal organs from changes in temperature and humidity, ultraviolet rays, pollutants, and external stimuli, and plays an important role in maintaining biological homeostasis such as body temperature control.
  • excessive stimulation from the outside, stress, nutritional deficiencies, hormonal abnormalities in the body, genetic diseases, etc. reduce the normal function of the skin, cause skin pigmentation, or promote skin aging phenomena such as loss of elasticity, keratinization, and wrinkle formation. Accordingly, efforts have been made to effectively suppress skin aging by maintaining skin-specific functions and skin-whitening active substances that can reduce hyperpigmentation of the skin.
  • the main causes of wrinkle formation in the epidermis are keratosis due to a decrease in turn-over of keratinocytes, a decrease in water-containing capacity, and keratin hardening, and the main factors in the hyperpigmentation of the skin are melanin pigment synthesis abnormalities and It is due to abnormal distribution.
  • the main function of melanin is to absorb UV light energy and protect cells from UV damage.
  • the production of melanin is converted into dopaquinone by the action of tyrosinase in melanocytes, followed by the action of enzymes and spontaneous oxidation reactions.
  • abnormally low production of melanin causes skin lesions such as vitiligo.
  • excessive synthesis of melanin by ultraviolet rays causes damage to the skin, as well as skin blackening, blemishes, and freckles, and furthermore, it may cause skin cancer. do.
  • Rhodiola Sachalinensis A Bor is a perennial medicinal plant belonging to the genus Pseudomonas phyllumaceae stone flowers and is distributed in the alpine regions of western and northern Asia. It is a plant with special adaptability that can survive at 1,700-2,300m above sea level, with low temperature, dryness, low oxygen, strong ultraviolet rays, and a large temperature difference between day and night. It is a plant with special adaptability to poor environmental factors (physical, chemical, biological factors). It is reported that it improves adaptive capacity and regulates physiological functions of organisms to maintain normal life activities, and has high antioxidant activity.
  • Rhododendron is used for edible and medicinal purposes, and has been used as a sedative, antipyretic, and astringent in the private sector.
  • oriental medicine it is effective in tonic medicine, especially for senile heart atrophy and sound stomach. It is used as a medicine to treat weakness and postpartum.
  • As a pharmacological ingredient it is reported that there are 20 kinds of amino acids and 18 kinds of trace elements along with the main four ingredients such as salidroside, rosavin, rosin and rosarin. Has become.
  • the Korea Ministry of Food and Drug Safety recognizes Honggyeongcheon as a health functional food ingredient with the effect of improving fatigue due to stress.
  • a cosmetic composition (Korean Patent No. 10-1661545) containing a fermented extract of honggyeongcheon with whitening activity, fermented by fermenting honggyeongcheon with mycelium of Cordyceps sinensis , Paecilomyces japonica . It relates to a method for preparing a cosmetic composition for skin whitening or wrinkle improvement comprising an extract as an active ingredient, and the fermentation mycelium used for biotransformation is different from the present invention, and there is a large difference in activity according thereto.
  • an object of the present invention is to provide a composition capable of improving skin moisturization, improving skin elasticity, improving skin wrinkles, or preventing or improving skin photoaging.
  • the present invention provides a composition for preventing or improving skin aging, containing as an active ingredient a fermented honggyeongcheon extract obtained after fermenting honggyeongcheon extract using glutinous rice cake mushroom.
  • the present invention also includes the steps of: (a) extracting honggyeongcheon roots using a low-carbon alcohol solvent; And (b) fermenting the extract obtained in step (a) using glutinous rice cake mushroom ( Bovista plumbea ).
  • the honggyeongcheon extract obtained by simply extracting honggyeongcheon as a solvent exhibits effective efficacy in vitro, but when it is applied to skin cosmetics or quasi-drugs, the effect is not substantially satisfactory.
  • the present inventors confirmed that the main ingredient showing efficacy in the honggyeongcheon extract was a polyphenol compound, and as a result of confirming the content of the polyphenolic compound after fermenting the honggyeongcheon extract with various mushroom mycelium, Honggyeongcheon fermented with glutinous rice cake mushroom mycelium It was confirmed that the extract fermentation broth had the most excellent effect of increasing the total phenol content and total flavonoid content compared to the fermented broth of honggyeongcheon extract fermented with honggyeongcheon extract or other types of mushroom mycelium.
  • the fermented broth of honggyeongcheon extract of the present invention exhibits the effect of scavenging free radicals, promoting collagen biosynthesis, inhibiting the expression and activity of collagenase (MMP-1), and producing melanin pigment. Since the inhibitory effect is excellent, it can be usefully used as an active ingredient of a composition for preventing or improving skin aging.
  • the present invention provides a composition for preventing or improving skin aging, containing as an active ingredient a fermented solution of honggyeongcheon extract obtained after fermenting honggyeongcheon extract using glutinous rice cake mushroom.
  • “Honggyeongcheon ( Rhodiola Sachalinensis ) of the present invention belongs to the Pseudomonas plant family Dolphinaceae, of course, including subspecies and varieties thereof.
  • the honggyeongcheon of the present invention is obvious in the art and is the same as the prevention and improvement object of the present invention Or, it includes a cohort herb that can be used for a similar purpose.
  • honggyeongcheon extract fermentation broth refers to a fermentation broth obtained after fermenting the extract of honggyeongcheon ( Rhodiola Sachalinensis ) using glutinous rice cake mushroom ( Bovista plumbea ).
  • the honggyeongcheon extract according to the present invention can be extracted from the roots of honggyeongcheon using an ultrasonic extractor using a low-carbon alcohol solvent.
  • the rhododendron roots may be dried roots generally sold on the market or powders thereof.
  • the roots of honggyeongcheon can be minced and dried to make them directly.
  • methanol, ethanol, propanediol, and a mixed solvent thereof may be used, and ethanol is preferably used.
  • ethanol is used as an extraction solvent to obtain a rhododendron extract, even if water is added to the extract, the elution or precipitation of the active ingredient does not occur, so there is an advantage that various subsequent processes can be applied.
  • the primary extract obtained by extracting the honggyeongcheon roots using a low-carbon alcohol solvent as described above may be further fractionated using an organic solvent having a different polarity.
  • a further purified fraction may be obtained by using various chromatography and reverse osmosis-membrane separation. Therefore, in the present invention, "honggyeongcheon extract” is a concept including all extracts, fractions and purified products obtained in each step of extraction, fractionation, or purification, and their dilutions, concentrates or dried products.
  • the glutinous rice cake mushroom used in the present invention ( Bovista plumbea ) is 2-4cm in size, somewhat spherical, and mainly grows in Europe and California. It is used for food, but its efficacy is not yet known in detail.
  • the glutinous rice cake mushroom belongs to the wrinkle mushroom family, and subspecies and varieties thereof are of course included.
  • the fermented solution of honggyeongcheon extract produced by fermentation with glutinous rice cake mushrooms as described above may be contained in a range of 0.001 to 50% by weight based on the total weight of the composition of the present invention. If it is less than 0.001% by weight, it is difficult to expect a substantial skin moisturization improvement, skin elasticity improvement, skin wrinkle improvement, or skin photoaging improvement effect of the composition, and if it exceeds 50% by weight, it is not economical because there is little synergistic effect due to excessive administration.
  • the composition of the present invention is used to improve skin wrinkles or skin photoaging.
  • the fermented broth of honggyeongcheon extract which is an active ingredient in the composition of the present invention, exhibits excellent anti-wrinkle and skin photoaging improvement effects by promoting collagen biosynthesis and inhibiting the expression or activity of collagenase (MMP-1) while having low cytotoxicity. Examples 5 and 6).
  • the composition of the present invention has the use of skin whitening or skin photoaging improvement.
  • whitening refers to an action of improving skin troubles by preventing blackening of the skin caused by deposition of melanin pigment.
  • the fermented broth of honggyeongcheon extract which is an active ingredient in the composition of the present invention, exhibits a whitening effect through inhibition of melanin production, and has high safety, so there is little side effects such as inducing skin irritation (Experimental Example 4).
  • skin aging prevention or improvement is interpreted as including skin moisturizing, skin elasticity improvement, wrinkle prevention, wrinkle removal, skin blackening prevention, skin pigmentation improvement, and skin aging inhibition through antioxidant action.
  • the composition of the present invention is a cosmetic composition.
  • the cosmetic composition of the present invention may include ingredients commonly used in cosmetic compositions in addition to the fermented solution of the honggyeongcheon extract as an active ingredient.
  • ingredients commonly used in cosmetic compositions for example, purified water, moisturizers, oils, surfactants, higher alcohols, thickeners, chelating agents, pigments, fatty acids, antioxidants, preservatives, waxes, pH regulators, and may include conventional adjuvants and carriers such as fragrances, but are limited thereto. no.
  • glycerin propylene glycol, butylene glycol, hexylene glycol, diglycerin, sorbitol, betaine, glyceres-26, and the like may be used.
  • the oils include mineral oil, cyclomethicone, squalane, octyldodecyl myristate, olive oil, macadamia nut oil, glyceryl octanoate, castor oil, ethylhexyl isononanoate, dimethicone, cyclopentasiloxane, Sunflower seed oil or the like may be used.
  • the surfactants include sorbitan sesquinoleate, polysorbate 60, glyceryl stearate, lipophilic glyceryl stearate, sorbitan oleate, sorbitan stearate, die-cetyl phosphate, sorbitan stearate/sugar Crosscocoate, glyceryl stearate/polyethylene glycol-100 stearate, ceteareth-6 olivate, rachidyl alcohol/behenyl alcohol/arachidyl glucoside, polypropylene glycol-26-butes-26/ Polyethylene glycol-40 hydrogenated castor oil or the like may be used.
  • xanthan gum carbomer, magnesium aluminum silicate, cellulose gum, dextrin palmitate, and the like may be used.
  • disodium EDTA As the chelating agent, disodium EDTA, tetrasodium EDTA, and the like may be used.
  • Blue No. 1, Red No. 40, Yellow No. 4, and Yellow No. 5 may be used as the pigment.
  • fatty acid stearyl acid, myristic acid, palmitic acid, isostearic acid, lauric acid, and the like may be used.
  • Tocopheryl acetate, butylated hydroxytoluene, and the like may be used as the antioxidant.
  • methylparaben, butylparaben, propylparaben, phenoxyethanol, imidazolidinylurea, chlorphenesin, and the like may be used.
  • wax carnauba wax, candenilla wax, beeswax, or brazing wax may be used.
  • Triethanolamine, citric acid, and the like may be used as the pH adjusting agent.
  • fragrance natural fragrances, combination fragrances, and the like may be used.
  • the cosmetic composition of the present invention can be prepared in any formulation conventionally prepared in the art.
  • it may be formulated as a solution, a suspension, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant-containing cleansing, an oil, a powder foundation, an emulsion foundation, a wax foundation, and a spray, etc. no.
  • it may be prepared in the form of a flexible lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, cleansing cream, cleansing water, pack, spray or powder.
  • the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide may be used as carrier components.
  • animal oil vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide
  • talc a paste, cream or gel
  • animal oil vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide
  • lactose When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as a carrier component.
  • lactose talc
  • silica aluminum hydroxide
  • calcium silicate or polyamide powder
  • a carrier component in particular, in the case of a spray, additionally chlorofluorohydrocarbon, propane , Butane or dimethyl ether.
  • a solvent, a solubilizing agent or an emulsifying agent is used as a carrier component, and examples are water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.
  • the formulation of the present invention is a suspension
  • a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, and crystallites Sex cellulose, aluminum metahydroxide, bentonite, agar or tracant, and the like
  • a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester
  • crystallites Sex cellulose aluminum metahydroxide, bentonite, agar or tracant, and the like
  • the carrier components include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide.
  • Ether sulfates, alkylamidobetaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters may be used.
  • the composition of the present invention may be prepared as a pharmaceutical composition.
  • the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier.
  • Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are commonly used in formulation, and are lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium dioxide, alginate, gelatin, silicic acid. Calcium, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, mineral oil, etc. It is not.
  • the pharmaceutical composition of the present invention may further include a emulsifying agent, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, and a preservative.
  • composition of the present invention may be administered orally or parenterally, preferably parenterally, more preferably, applied by topical administration by application.
  • the appropriate dosage of the pharmaceutical composition of the present invention is prescribed in various ways depending on factors such as the formulation method, the mode of administration, the patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate and response sensitivity. Can be.
  • the dosage of the pharmaceutical composition of the present invention is in the range of 0.001 to 100 mg/kg on an adult basis.
  • the pharmaceutical composition of the present invention is prepared in unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person having ordinary knowledge in the technical field to which the present invention belongs. Or it can be made by incorporating it into a multi-dose container.
  • the formulation may be in the form of a solution, suspension, syrup, or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, powder, granule, tablet or capsule, and may additionally include a dispersant or a stabilizer.
  • the pharmaceutical composition of the present invention when it is an external preparation for skin, it may be prepared in one formulation selected from the group consisting of cream, gel, ointment, liquid, powder, and spray.
  • composition of the present invention can be prepared as a food composition.
  • the composition of the present invention when prepared as a food composition, in addition to the fermentation broth of honggyeongcheon extract as an active ingredient, it may include ingredients that are commonly added during food production.
  • it may include proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents.
  • carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose, sucrose, and oligosaccharides; And common sugars such as polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and fondhritol.
  • the flavoring agent natural flavoring agents and synthetic flavoring agents may be used.
  • the food composition of the present invention when the food composition of the present invention is prepared as a drink, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice in addition to the fermented solution obtained after fermenting Honggyeongcheon extract using glutinous rice cake mushroom as an active ingredient of the present invention , Duchung extract, jujube extract, licorice extract, etc. may be additionally included.
  • the rhododendron root may use a powder generally sold on the market.
  • the roots of honggyeongcheon may be dried until the moisture content is 8 to 15% by weight, and then pulverized.
  • methanol, ethanol, propanediol, and a mixed solvent thereof may be used as the low-carbon alcohol solvent, and ethanol is preferably used.
  • the extract of honggyeongcheon may be fermented by mixing the mycelium of glutinous rice cake mushroom ( Bovista plumbea ) or the entire culture thereof, followed by main culture.
  • the pH of the culture medium is 5.0-7.0
  • composition containing the fermentation broth of honggyeongcheon extract of the present invention as an active ingredient promotes collagen synthesis and inhibits the expression and activity of MMP-1, a collagen-degrading enzyme, thereby preventing skin aging and wrinkle formation, and has an inhibitory effect on melanin pigment production. As it is excellent, it is effective in improving skin whitening and skin blackening caused by photoaging.
  • Glutinous rice cake mushroom mycelium was cultured on a slope in a test tube containing potato dextrose agar medium, stored at 4°C, and subcultured every 1 month before use.
  • the mycelium passaged in the slope medium was homogenized aseptically, and inoculated so that 5% (v/v) mycelium of glutinous rice cake mushrooms was inoculated in the mixed medium obtained in Comparative Example 1 with the extract of honggyeongcheon root extract and yeast-wort.
  • glutinous rice cake mushroom mycelium was removed from the culture broth, and then a fermented broth of Honggyeongcheon extract was obtained.
  • This experimental example measured the scavenging effect on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical by slightly modifying the method used by Yoshida et al. (1989) to measure the antioxidant effect of the material obtained in Example 1. . Since DPPH is a stable free radical and exhibits purple color, it measures the degree to which purple is removed as the sample scavenges the radicals.
  • DPPH was dissolved in methanol at a concentration of 1 ⁇ 10 -4 M, and 0.15 ml of this solution was put into a test tube. 0.15 ml of a sample of each concentration was added thereto, and after reacting at room temperature for 20 minutes, absorbance was measured at 565 nm.
  • vitamin C was used instead of the material of the examples.
  • the free radical scavenging effect of each sample was expressed as SC 50 by calculating the concentration at which 50% of the radicals were scavenging. At this time, as a negative control, the reaction absorbance of the solution without sample treatment was used.
  • the positive control vitamin C had an SC50 value of 1.78, whereas the fermented broth of honggyeongcheon extract of the present invention was slightly higher at 12.6. However, it is included in the concentration of DPPH radical scavenging activity, which is very potent compared to the activity of extracts other than a single substance.
  • This experimental example was used by modifying the method of Roberta et al. in order to measure the ABTS radical scavenging activity of the material obtained in Example 1.
  • 7mM ABTS and 2.45mM potassium sulfate were added to distilled water to produce ABTS cations for 16 hours at room temperature. At this time, it was prepared by diluting so that the value of absorbance at 734 nm was 0.7 or less.
  • 50 ul of each concentration sample was added to 950 ul of the ABTS solution and reacted for 10 minutes, and the absorbance was measured at 734 nm.
  • BHA was used instead of the material of the examples.
  • the free radical scavenging effect of each sample was expressed as SC 50 by calculating the concentration at which 50% of the radicals were scavenging. At this time, as a negative control, the reaction absorbance of the solution without sample treatment was used.
  • the BHA which is a positive control, had an SC50 value of 1.52, whereas the fermented broth of honggyeongcheon extract of the present invention was 3.75. This was found to be a somewhat higher concentration than the positive control BHA, but, like the DPPH radical scavenging activity, it is included in the very strong ABTS radical scavenging activity concentration compared to the activity of other extracts other than a single substance.
  • mice melanocytes were used to observe the inhibitory effect on melanin pigment production of the samples obtained in Example 1. That is, Melan A cells were dispensed into a 6-well plate at 2 ⁇ 10 5 /well, and the culture solution was removed for 18 hours under conditions of 37°C and 5% CO 2 incubator. At this time, the positive control arbutin and the sample were each added and cultured. After incubation for 48 hours, the medium was removed, and the bottom of the 6-well was washed with PBS (phosphate buffered saline) evenly, and the cells were collected in a micro tube by treatment with trypsin-EDTA.
  • PBS phosphate buffered saline
  • the recovered cells were added with 200 ⁇ L of 1 N NaOH and left in a water bath at 80° C. for 1 hour to dissolve the melanin in the cells, and then left on ice for 1 hour, and the melanin content was measured at 405 nm using ELISA. Each data is expressed by correcting the amount of protein.
  • DMEM Dulbecco's Modified Eagle's Media
  • fetal calf serum 2.5% fetal calf serum and human fibroblasts (5000 cells/well)
  • fetal calf serum 2.5% fetal calf serum
  • human fibroblasts 5000 cells/well
  • the collected cell culture was measured for collagen synthesis using a collagen protein measuring instrument (Catalog N.: MK 101, Takara Shuzo Co. Ltd, Japan).
  • the collected cell culture solution was put into a 96 hole plate incubator uniformly coated with the primary collagen antibody, and an antigen-antibody reaction was performed for 3 hours. After 3 hours, the secondary collagen antibody to which the chromophore was bound was placed in a 96 hole plate incubator and reacted for 15 minutes. After 15 minutes, the color development inducing substance was added to develop color at room temperature, and 1M sulfuric acid was added to stop the color development. The color of the reaction color is yellow, and its intensity varies depending on the degree of reaction. The absorbance was measured at 450 nm using a yellowish 96 hole plate incubator, and the degree of collagen synthesis was calculated by Equation 1 below. At this time, the reaction absorbance of the cell culture solution not treated with the sample was used as a control.
  • Collagen synthesis effect (%) [(reaction absorbance of control group-reaction absorbance of cell culture solution of sample)/reaction absorbance of control group] ⁇ 100
  • UVA was irradiated to human dermal fibroblasts using an ultraviolet chamber with an energy of 6.3 J/cm 2 , exchanged for a medium to which a sample was added, and cultured for 24 hours, and then the medium was recovered and coated on a 96 hole plate incubator.
  • the primary antibody (MMP-1(Ab-5) monoclonal antibody) was treated and reacted at 37° C. for 90 minutes, and then 3% BSA was treated for 1 hour to block unbound sites.
  • alkaline phosphatase substrate solution (1 mg/ml p-nitrophenyl phosphate in diethanolamine buffer) was added and reacted at room temperature for 30 minutes. Absorbance was measured at 405 nm using an absorbance photometer. The degree of inhibition of MMP-1 activity was calculated by Equation 2 below. At this time, the reaction absorbance of the cell culture solution not treated with the sample was used as a control.
  • MMP-1 expression inhibition rate (%) [(reaction absorbance of control group-reaction absorbance of cell culture solution of sample)/reaction absorbance of control group] ⁇ 100
  • the fermented broth of honggyeongcheon extract of the present invention exhibited a higher inhibitory effect on the expression of collagenase than 100 ⁇ M of hydrogen peroxide (H 2 O 2 ) as a positive control.
  • a cosmetic composition containing the fermented liquid of honggyeongcheon extract obtained in Example 1 was prepared to evaluate the effect of improving elasticity recovery, moisturizing, and skin density in humans.
  • the cosmetic used in the efficacy evaluation is in the form of a cream, and its composition is as shown in [Table 7].
  • the b) phase recorded in [Table 7] was heated and stored at 70°C. After pre-emulsification by adding phase a) to the b) phase, uniformly emulsified with a homomixer, and then gradually cooled to cream (Example 2 ) was prepared
  • the experimenter (30-50 years old female) subject to 21 subjects, the experimenter directly applied the cream prepared in Example 2 twice a day for 2 consecutive months.
  • the effect of improving elasticity recovery, improving mid- to long-term moisturizing and improving skin density was measured by measuring the elasticity of the epidermis, the amount of moisture in the stratum corneum, and the density of the dermis before using the product, after 2 weeks, and after 4 weeks of use.
  • [Table 8] is a measurement of the skin elasticity of the experimenter using the cream prepared in Example 2.
  • the skin elasticity of the experimenter who applied the cream containing the fermented honggyeongcheon extract increased by 9.43% after 2 weeks of the test and 16.67% after 4 weeks of the test, resulting in a statistically significant level of epidermal elasticity compared to before the test. Increased. Therefore, it is believed that a cream containing fermented rhododendron root extract can help improve epidermal elasticity.
  • [Table 9] is a measurement of the moisture content of the stratum corneum of the experimenter using the cream prepared in Example 2.
  • the amount of water in the epidermis of the experimenter who applied the cream containing the fermented honggyeongcheon extract increased by 7.43% after 2 weeks of the test and 16.28% after 4 weeks of the test, which was statistically significant in the stratum corneum than before the test.
  • the moisture content increased. Therefore, it is thought that a cream containing fermented rhododendron root extract can help improve the moisture content in the stratum corneum.
  • [Table 10] is a measurement of the dermal density of the experimenter using the cream prepared in Example 2.
  • the dermal density of the experimenter who applied the cream containing the fermented honggyeongcheon extract increased 12.58% from 2 weeks after the test and 24/67% after 4 weeks of the test, to a statistically significant level compared to before the test.
  • the dermal density increased. Therefore, it is thought that a cream containing fermented rhododendron root extract can help improve skin density.

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Abstract

La présente invention concerne une composition pour prévenir ou atténuer le vieillissement cutané, contenant, en tant que principe actif, un liquide de fermentation d'extrait de Rhodiola Sachalinensis obtenu après la fermentation d'un extrait de Rhodiola Sachalinensis à l'aide de Bovista plumbea. Une composition de la présente invention peut prévenir le vieillissement cutané et la formation de rides en favorisant la biosynthèse de collagène et en inhibant l'expression et l'activité de MMP-1, qui est une collagénase, et présente un excellent effet d'inhibition de la production du pigment qu'est la mélanine, ce qui est efficace pour le blanchiment de la peau et pour l'atténuation de l'apparition de taches brunes sur la peau provoquée par le photovieillissement.
PCT/KR2020/007128 2019-06-03 2020-06-02 Composition pour prévenir ou atténuer le vieillissement cutané, contenant un extrait de rhodiola sachalinensis fermenté à l'aide de bovista plumbea WO2020246766A1 (fr)

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KR102512796B1 (ko) * 2021-02-08 2023-03-24 이주운 항산화성이 증진된 몰로키아 추출물 및 이의 용도
KR20230138679A (ko) 2022-03-24 2023-10-05 이주운 홍경천 발효물을 포함하는 인지기능 장애 또는 기억력 장애 예방 또는 개선용 조성물

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