US20150104529A1 - Cosmetic composition containing fermented ginseng berry pleurotus ferulae product - Google Patents

Cosmetic composition containing fermented ginseng berry pleurotus ferulae product Download PDF

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US20150104529A1
US20150104529A1 US14/250,136 US201414250136A US2015104529A1 US 20150104529 A1 US20150104529 A1 US 20150104529A1 US 201414250136 A US201414250136 A US 201414250136A US 2015104529 A1 US2015104529 A1 US 2015104529A1
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ginseng berry
cosmetic composition
product
fermented ginseng
skin
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Kyung Rok Lee
Il Hong
Do Gyeong Lee
Sung Min Park
Jung No Lee
Nu Rim Lee
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AMI COSMETIC Co Ltd
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AMI COSMETIC Co Ltd
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Assigned to AMI COSMETIC CO., LTD. reassignment AMI COSMETIC CO., LTD. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: HONG, IL, LEE, DO GYEONG, LEE, JUNG NO, LEE, KYUNG ROK, LEE, NU RIM, PARK, SUNG MIN
Publication of US20150104529A1 publication Critical patent/US20150104529A1/en
Priority to US15/016,495 priority Critical patent/US10350156B2/en
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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
    • A61K8/18—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
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • 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
    • A61Q19/007—Preparations for dry 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
    • A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
    • C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
    • 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
    • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
    • A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
    • 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 cosmetic composition containing, as an active ingredient, a fermented ginseng berry Pleurotus ferulae product. More specifically, the present invention relates to a cosmetic composition containing, as an active ingredient, a fermented product prepared by fermenting a ginseng berry extract with Pleurotus ferulae mycelium.
  • Aging of the skin is classified into two types, i.e., intrinsic (chronological) aging and photoaging [Gilchrest B A: J. Am. Acad. Dermatol., 21, 610-613 (1989)].
  • Intrinsic aging naturally occurs due to decrease in physiologic functions with age [Braverman I M, et al.: J. Invest. Dermatol., 78, 434-443 (1982)].
  • Photoaging means change associated with appearance and functions of the skin, caused by repeated exposure of the skin to solar radiation [Ridder G M et al.: J. Am. Acad. Dermatol., 25, 751-760 (1991)].
  • aging of the skin may be caused by ultraviolet radiation, stress, disease conditions, environmental factors, wounds and activation of active oxygen species with age. As such conditions worsen, antioxidant defense mechanisms present in vivo are destructed, cells and tissues are damaged, and adult diseases and aging are facilitated. More specifically, oxidation of lipids, proteins, polysaccharides, nucleic acids and the like, which are primary components of the skin, and destruction of skin cells and tissues thus causes aging of the skin.
  • oxidation of proteins involves cleavage of collagen, hyaluronic acid, elastin, proteoglycan, fibronetin and the like, which are connective tissues of the skin, results in over-inflammation, causes damage to elasticity of the skin and, in serious cases, brings about mutations caused by DNA modification, emergence of cancers and deterioration in immune function.
  • MMP matrix metalloproteinase
  • MMPs matrix metalloproteinases
  • melanin is produced through conversion of tyrosine into dopa, dopaquinone and then dopachrome by actions of tyrosinase present in pigment cells.
  • Melanin is present in the skin, which protects the body from ultraviolet radiation and has an essential function on control of hormone secretion in vivo.
  • over-production of melanin is known to create spots, freckles and the like, accelerate aging of the skin and play an important role in inducing skin cancers. As such, research and development to prevent melanin over-production is actively underway.
  • Ascorbic acid Japanese Patent Publication Sho. 4-9320
  • hydroquinone Japanese Patent Publication Sho.
  • human skin is an organ which serves as a protective membrane which protects human from external environments, and functions to prevent loss of biogenic substances such as water and electrolytes and prevent harmful substances from invading into the human.
  • Skin is broadly divided into the epidermis, the dermis and subcutaneous fat.
  • the epidermis is composed of keratinocytes and melanocytes.
  • a keratinocyte layer which constitutes the outermost layer of the skin directly contacts external environments and thus should have high physical or chemical resistance or superior barrier property to materials, prevent release (loss) of water to the outside from the human, and maintain flexibility due to presence of a proper amount of water therein.
  • TEWL trans-epidermal water loss
  • a hydrophilic substance capable of retaining water called “natural moisturizing factor (NMF)”, present in the keratinocyte layer is known to play an important role in moisturizing the skin.
  • NMF natural moisturizing factor
  • a normal keratinocyte layer maintains a water content of about 10 to about 30%, the skin becomes smooth and soft and normally exerts the function of protecting the body.
  • the water content of the keratinocyte layer is 10% or less, the skin becomes rough, losses its body protection function and is aged.
  • a scaling phenomenon in which scale-like keratinocytes are peeled off the surface of the skin due to weak adhesion between keratinocytes occurs.
  • the dryness of the skin is due to the fact that the dry skin has a water content of the keratinocyte layer lower than that of normal skin.
  • even healthy skin has bad conditions due to lack of water caused by exposure to harsh external environments, i.e., wind, cold weather, sunlight, washing or shaving.
  • cosmetics containing components similar to sebum, NMF components or moisturizers such as polyols were used.
  • glycerin or sorbitol having three or more hydroxyl groups (OH groups) as water-soluble polyols, exhibit excellent moisturizing effect, but renders discomfort upon use due to severe stickiness, and propylene glycol, 1,3-butylene glycol and the like having two hydroxyl groups (OH groups) cause side effects to the skin.
  • the water retention property of the keratinocyte layer may be controlled by natural moisturizing factors (NMFs) composed of amino acids, lactic acid, urea and inorganic salts.
  • NMFs natural moisturizing factors
  • an extract of ginseng berries prepared by fermentation with Pleurotus ferulae mycelium exhibits anti-oxidation, anti-inflammation, collagen synthesis promotion, skin wrinkle care, whitening, moisturizing, skin barrier function and atopy alleviation effects.
  • the present invention was completed based on this discovery.
  • Patent Document Korean Patent No. 10-0887631 (registered on Mar. 2, 2009) discloses a composition for external application to the skin containing a ginseng berry extract and a cosmetic containing the ginseng berry extract as an active ingredient.
  • the present invention has been made in view of the above problems, and it is an object of the present invention to develop and provide a cosmetic composition using a ginseng berry which is applicable to skin cosmetics and has considerably safety due to non-harmness to the human body.
  • a cosmetic composition containing, as an active ingredient, a fermented ginseng berry Pleurotus ferulae product obtained by fermenting a ginseng berry extract with Pleurotus ferulae.
  • the cosmetic composition of the present invention is for example used for any one selected from anti-oxidation, anti-inflammation, skin wrinkle care, skin whitening, skin moisturizing and atopy alleviation.
  • the ginseng berry extract is for example extracted using any one extraction solvent selected from the group consisting of water, C1-C4 anhydrous or aqueous lower alcohol, acetone, ethyl acetate, butyl acetate and 1,3-butylene glycol.
  • the fermented ginseng berry Pleurotus ferulae product is preferably present in an amount of 0.0001 to 100.0% by weight, with respect to the total weight of the cosmetic composition.
  • FIG. 1 is a graph showing comparison in free radical scavenging activity at different concentrations (0.001%, 0.002%, 0.005%, 0.01%) between a ginseng berry extract obtained in Preparation Example 1 and fermented ginseng berry products obtained in Example 1 and Comparative Examples 1 to 4, wherein ‘EGCG’ represents epigallocatechin gallate, ‘a’ represents a ginseng berry extract of Preparation Example 1, ‘b’ represents a fermented ginseng berry Pleurotus ferulae product of Example 1, ‘c’ represents a fermented ginseng berry Tricholoma matsutake product of Comparative Example 1, ‘d’ represents a fermented ginseng berry Sarcodon aspratus product of Comparative Example 2, ‘e’ represents a fermented ginseng berry Phellinus linteus product of Comparative Example 3, and ‘f’ represents a fermented ginseng berry Ganoderma lucidum product of Comparative Example 4;
  • EGCG represents
  • FIG. 2 is a graph showing comparison in 5-lipoxygenase inhibition activity at different concentrations (0.001%, 0.002%, 0.005%, 0.01%) between the ginseng berry extract obtained in Preparation Example 1 and the fermented ginseng berry products obtained in Example 1 and Comparative Examples 1 to 4, wherein ‘NDGA’ represents nordihydroguaiaretic acid, ‘a’ represents a ginseng berry extract of Preparation Example 1, ‘b’ represents a fermented ginseng berry Pleurotus ferulae product of Example 1, ‘c’ represents a fermented ginseng berry Tricholoma matsutake product of Comparative Example 1, ‘d’ represents a fermented ginseng berry Sarcodon aspratus product of Comparative Example 2, ‘e’ represents a fermented ginseng berry Phellinus linteus product of Comparative Example 3, and ‘f’ represents a fermented ginseng berry Ganoderma lucidum product of Comparative Example 4;
  • FIG. 3 is a graph showing comparison in collagen synthesis at different concentrations (0.001%, 0.002%, 0.005%, 0.01%) between the ginseng berry extract obtained in Preparation Example 1 and the fermented ginseng berry products obtained in Example 1 and Comparative Examples 1 to 4, wherein ‘L-AA’ represents L-ascorbic acid, ‘a’ represents a ginseng berry extract of Preparation Example 1, ‘b’ represents a fermented ginseng berry Pleurotus ferulae product of Example 1, ‘c’ represents a fermented ginseng berry Tricholoma matsutake product of Comparative Example 1, ‘d’ represents a fermented ginseng berry Sarcodon aspratus product of Comparative Example 2, ‘e’ represents a fermented ginseng berry Phellinus linteus product of Comparative Example 3, and ‘f’ represents a fermented ginseng berry Ganoderma lucidum product of Comparative Example 4;
  • FIG. 4 is a graph showing comparison in MMP-1 inhibition activity at different concentrations (0.001%, 0.002%, 0.005%, 0.01%) between the ginseng berry extract obtained in Preparation Example 1 and the fermented ginseng berry products obtained in Example 1 and Comparative Examples 1 to 4, wherein ‘retinol’ represents retinol, ‘a’ represents a ginseng berry extract of Preparation Example 1, ‘b’ represents a fermented ginseng berry Pleurotus ferulae product of Example 1, ‘c’ represents a fermented ginseng berry Tricholoma matsutake product of Comparative Example 1, ‘d’ represents a fermented ginseng berry Sarcodon aspratus product of Comparative Example 2, ‘e’ represents a fermented ginseng berry Phellinus linteus product of Comparative Example 3, and ‘f’ represents a fermented ginseng berry Ganoderma lucidum product of Comparative Example 4; and
  • FIG. 5 is a graph showing comparison in inhibition of tyrosinase activity at different concentrations (0.001%, 0.002%, 0.005%, 0.01%) between the ginseng berry extract obtained in Preparation Example 1 and the fermented ginseng berry products obtained in Example 1 and Comparative Examples 1 to 4, wherein ‘Kojic acid’ represents kojic acid, ‘a’ represents a ginseng berry extract of Preparation Example 1, ‘b’ represents a fermented ginseng berry Pleurotus ferulae product of Example 1, ‘c’ represents a fermented ginseng berry Tricholoma matsutake product of Comparative Example 1, ‘d’ represents a fermented ginseng berry Sarcodon aspratus product of Comparative Example 2, ‘e’ represents a fermented ginseng berry Phellinus linteus product of Comparative Example 3, and ‘f’ represents a fermented ginseng berry Ganoderma lucidum product of Comparative Example 4.
  • Panax ginseng C. A. Meyter used for the present invention which is called “ginseng berry” is the fruit of the 4 years or more old ginseng plant harvested for only about one week in the middle of July.
  • Ginseng berry extracts have been widely utilized in a variety of applications to date. There is no research demonstrating the fact discovered by the present invention that the extract of ginseng berries fermented with Pleurotus ferulae exhibits considerably superior anti-oxidation, anti-inflammation, collagen synthesis facilitation, skin wrinkle care, whitening, moisturizing, skin barrier improvement and atopy alleviation effects, as compared to ginseng berry extracts.
  • Pleurotus ferulae Lenzi is a kind of mushroom which belongs to pleurototaceae and pleurotus.
  • Pleurotus ferulae is the name given to call to grow in glassroots called “ ferula asafetida ” in Xinjiang, China and has been artificially cultivated since 1983.
  • Pleurotus ferulae is known to have been first found in the Madonic Mountain, the Sicily Island, Italy in 1963.
  • Pleurotus ferulae is known to be a nutrient repository.
  • Pleurotus ferulae having a delicate and soft pine fragrance had been used in China from old times due to potent medical efficacies including treatment of stomach and kidney disorders, cough drop, removal of inflammation and prevention of diseases of obstetrics and gynecology.
  • the present inventors fermented ginseng berries with a variety of mushrooms as raw materials.
  • the present inventors discovered that the ginseng berry fermented with Pleurotus ferulae mycelium exhibited superior anti-oxidation, anti-inflammation, collagen synthesis facilitation, skin wrinkle care, whitening, moisturizing, skin barrier improvement and atopy alleviation effects.
  • the present invention was completed based on this discovery.
  • the ginseng berry extract may be prepared using a common solvent in accordance with an ordinary method well-known in the art, that is, under ordinary temperature and pressure conditions.
  • the ginseng berry extract which is the active ingredient of the composition of the present invention may be prepared by extracting ginseng berries using a solvent selected from the group consisting of water, C1-C4 anhydrous or aqueous lower alcohol, acetone, ethyl acetate, butyl acetate and 1,3-butylene glycol, and optionally mixing the extract.
  • the solvent is preferably alcohol, more preferably ethanol, even more preferably, 70% ethanol.
  • the extraction solvent is preferably added in an amount of 1 to 15-fold, more preferably 5 to 10-fold, most preferably 7-fold, of a dry weight of the ginseng berry.
  • the fermented ginseng berry Pleurotus ferulae product according to the present invention may include an extract obtained by the extraction method described above and an extract obtained by a subsequent common purification process.
  • the fermented ginseng berry Pleurotus ferulae product includes fragments obtained by further performing a variety of purification processes such as separation using ultrafiltration membranes having a constant cut-off value and separation using a variety of chromatography (manufactured for separation according to size, charges, and hydrophobicity or hydrophilicity).
  • the fermented ginseng berry Pleurotus ferulae product according to the present invention may include a fermented product prepared in the form of a powder, obtained by a further process such as distillation under reduced pressure, and lyophilization or spray-drying.
  • the content of the fermented ginseng berry Pleurotus ferulae product in the cosmetic composition of the present invention is preferably 0.0001 to 100.0% by weight, more preferably 0.001 to 90.0% by weight, most preferably 0.1 to 85.0% by weight, with respect to the total weight of the cosmetic composition.
  • the fermented ginseng berry Pleurotus ferulae product according to the present invention superior anti-oxidation effect (see Experimental Example 1), superior anti-inflammatory effect (see Experimental Example 3), superior collagen synthesis effect (see Experimental Example 4), superior wrinkle alleviation effect (see Experimental Example 5), and superior whitening effect (see Experimental Example 6).
  • the cosmetic composition prepared using the fermented ginseng berry Pleurotus ferulae product having the effects described above, as an active ingredient exhibits superior skin moisturizing improvement effect (see Experimental Example 7), superior skin barrier improvement effect (see Experimental Example 8), and superior atopy alleviation effect (see Experimental Example 9).
  • the cosmetic composition according to the present invention may have any one formulation selected from the group consisting of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansings, oils, powder foundations, emulsion foundations, wax foundations and sprays, but the present invention is not necessarily limited thereto.
  • composition of the present invention may further include additives commonly used in cosmetics, such as hydrophilic or lyphophilic gelling agents, hydrophilic or lyphophilic activators, preservatives, antioxidants, solvents, flavoring agents, fillers, blockers, pigments, deodorant agents and dyes.
  • additives commonly used in cosmetics, such as hydrophilic or lyphophilic gelling agents, hydrophilic or lyphophilic activators, preservatives, antioxidants, solvents, flavoring agents, fillers, blockers, pigments, deodorant agents and dyes.
  • additives may be present in amounts commonly used in the art and may be for example 0.0001 to 100.0% by weight with respect to the total weight of the cosmetic composition.
  • the additives and contents thereof are selected such that preferred features of the cosmetic composition according to the present invention are not impaired.
  • Ginseng berries were washed with distilled water, dried, thoroughly crushed and passed through a 100 mesh sieve.
  • the crushed ginseng berry granules were added to 70% ethanol such that a concentration of the resulting mixture was adjusted to 150 g/L, extracted under reflux for 5 hours three times and then macerated. Then, the resulting product was filtered through Whatman #3 filter paper, concentrated under reduced pressure at 50° C. or less and lyophilized.
  • Ethanol was added to the fermented ginseng berry product obtained by fermentation and then primary removal of culture bacteria so as to obtain a final 70% (V/V) aqueous ethanol solution, extracted under reflux for 5 hours three times, macerated and filtered through Whatman #3 filter paper. After filtering, the resulting extract was concentrated in a concentrator under reduced pressure at 50° C. or less and lyophilized.
  • Comparative Example 1 the ginseng berry extract obtained in Preparation Example 1 was fermented with Tricholoma matsutake mycelium to prepare a fermented ginseng berry Tricholoma matsutake product.
  • the present experiment was performed in the same manner as in Example 1 except that Tricholoma matsutake was used, instead of Pleurotus ferulae.
  • Comparative Example 1 the ginseng berry extract obtained in Preparation Example 1 was fermented with Sarcodon aspratus mycelium to prepare a fermented ginseng berry Sarcodon aspratus product.
  • the present experiment was performed in the same manner as in Example 1 except that Sarcodon aspratus was used, instead of Pleurotus ferulae.
  • Comparative Example 1 the ginseng berry extract obtained in Preparation Example 1 was fermented with Phellinus linteus mycelium to prepare a fermented ginseng berry Phellinus linteus product.
  • the present experiment was performed in the same manner as in Example 1 except that Phellinus linteus was used, instead of Pleurotus ferulae.
  • Comparative Example 1 the ginseng berry extract obtained in Preparation Example 1 was fermented with Ganoderma lucidum mycelium to prepare a fermented ginseng berry Ganoderma lucidum product.
  • the present experiment was performed in the same manner as in Example 1 except that Ganoderma lucidum was used, instead of Pleurotus ferulae.
  • a powder of the ginseng berry extract obtained in Preparation Example 1 and powders of fermented ginseng berry products obtained in Example 1 and Comparative Examples 1 to 4 were suspended in distilled water and free radical scavenging activities thereof were evaluated at different concentrations (0.001%, 0.002%, 0.005%, 0.01%).
  • the DPPH (2,2-diphenyl-1-picrylhydrazyl) method is used to measure variation in color from absorbance at 540 nm when scavenging DPPH (2,2-Diphenyl-1-picrylhydrazyl) which is a radical whose inhibitors are stable).
  • Samples used for this experiment are shown in the following Table 1 and free radical scavenging activity was measured using the following Equation 1.
  • Epigallocatechin gallate (EGCG) was used as a control group.
  • Epigallocatechin gallate (EGCG, epigallocatechin-3-gallate) is a polyphenol extracted from green tea leaves, which has been found to be a potent antioxidant.
  • the fermented Pleurotus ferulae product (Example 1) exhibited the highest free radical scavenging activity. As free radical scavenging activity increases, antioxidative activity increases. Accordingly, the fermented ginseng berry Pleurotus ferulae product exhibits antioxidative activity higher than the ginseng berry extract.
  • ‘a’ represents a ginseng berry extract of Preparation Example 1
  • ‘b’ represents a fermented ginseng berry Pleurotus ferulae product of Example 1
  • ‘c’ represents a fermented ginseng berry Tricholoma matsutake product of Comparative Example 1
  • ‘d’ represents a fermented ginseng berry Sarcodon aspratus product of Comparative Example 2
  • ‘e’ represents a fermented ginseng berry Phellinus linteus product of Comparative Example 3
  • ‘f’ represents a fermented ginseng berry Ganoderma lucidum product of Comparative Example 4.
  • the ginseng berry extract powder obtained in Preparation Example 1 and fermented ginseng berry product powders obtained in Example 1 and Comparative Examples 1 to 4 were suspended in distilled water to obtain different concentrations (0.001%, 0.002%, 0.005%, 0.01%) of suspensions and cell survival rates of the suspensions were measured in accordance with the following method.
  • Cytotoxicty was measured by a Mosmann method which measures cell survival rate using a MTT ⁇ 3-(4,5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide ⁇ reagent.
  • HDF was seeded at a concentration of 1 ⁇ 10 4 cells/well on a 96-well plate and cultured under conditions of 37° C. and 5% CO 2 for 24 hours. After a culture medium was removed, the HDF was cultured in a medium treated with the sample at different concentrations for 24 hours, and the medium was removed and washed twice with phosphate buffered saline (PBS). MTT was dissolved at a concentration of 5 mg/mL in PBS, 50 L of the resulting MTT solution was added to HDF and HDF was cultured under the conditions of 37° C. and 5% CO 2 for 2 hours. 100 L of dimethyl sulfoxide (DMSO) was added to each well, followed by stirring for 10 minutes. Then, absorbance at 40 nm was measured.
  • DMSO dimethyl sulfoxide
  • the ginseng berry extract obtained in Preparation Example 1 and the fermented ginseng berry products obtained in Example 1 and Comparative Examples 1 to 4 were suspended in distilled water and anti-inflammatory effects thereof were evaluated by performing a 5-lipoxygenase inhibitory activity test under different concentration conditions (0.001%, 0.002%, 0.005%, 0.01%).
  • Lipoxygenase is an enzyme which produces various chemical mediators involved in inflammatory or allergic reaction in the body. Inhibitors of lipoxygenase cause inhibition of production of a variety of chemical mediators and are thus effective in alleviating allergies. That is, as 5-lipoxygenase inhibitory capacity increases, anti-inflammatory effect increases. In the present experimental example, activity of lipoxygenase may be indicated by peroxide production measured using substrates and enzymes.
  • the present experiment was performed using samples shown in the following Table 2 and 5-lipoxygenase inhibitory activity was measured using Equation 1.
  • the control group was nordihydroguaiaretic acid (NDGA) which is a potent antioxidant of fats and oils.
  • the fermented ginseng berry products of the present invention (Example 1 and Comparative Examples 1 to 4) exhibited superior 5-lipoxygenase inhibitory capacity to the ginseng berry extract (Preparation Example 1).
  • the fermented Pleurotus ferulae product (Example 1) exhibited the highest 5-lipoxygenase inhibitory capacity and the fermented ginseng berry Pleurotus ferulae product exhibited higher anti-inflammatory effects than the ginseng berry extract.
  • ‘a’ represents a ginseng berry extract of Preparation Example 1
  • ‘b’ represents a fermented ginseng berry Pleurotus ferulae product of Example 1
  • ‘c’ represents a fermented ginseng berry Tricholoma matsutake product of Comparative Example 1
  • ‘d’ represents a fermented ginseng berry Sarcodon aspratus product of Comparative Example 2
  • ‘e’ represents a fermented ginseng berry Phellinus linteus product of Comparative Example 3
  • ‘f’ represents a fermented ginseng berry Ganoderma lucidum product of Comparative Example 4.
  • the ginseng berry extract obtained in Preparation Example 1 and the fermented ginseng berry product powders obtained in Example 1 and Comparative Examples 1 to 4 were suspended in distilled water and a collagen biosynthesis increase at different concentrations (0.001%, 0.002%, 0.005%, 0.01%) was evaluated.
  • Human dermal fibroblasts were seeded on a 24-well plate at a concentration of 5 ⁇ 10 4 cells/well and cultured under the conditions of 37° C. and 5% CO 2 for 24 hours, and were further cultured in serum-free DMEMs (Dulbecco's Modified Eagle's Mediums) containing the ginseng berry extract obtained in Preparation Example 1 and the fermented ginseng berry products obtained in Example 1 and Comparative Examples 1 to 4 and in a serum-free DMEM medium not containing the fermented ginseng berry product and the ginseng berry extract, as a control group.
  • serum-free DMEMs Dulbecco's Modified Eagle's Mediums
  • the fermented ginseng berry products of the present invention (Example 1 and Comparative Examples 1 to 4) exhibited higher collagen synthesis at all the concentrations than the ginseng berry extract (Preparation Example 1).
  • the fermented Pleurotus ferulae product (Example 1) exhibited the highest collagen synthesis.
  • ‘a’ represents a ginseng berry extract of Preparation Example 1
  • ‘b’ represents a fermented ginseng berry Pleurotus ferulae product of Example 1
  • ‘c’ represents a fermented ginseng berry Tricholoma matsutake product of Comparative Example 1
  • ‘d’ represents a fermented ginseng berry Sarcodon aspratus product of Comparative Example 2
  • ‘e’ represents a fermented ginseng berry Phellinus linteus product of Comparative Example 3
  • ‘f’ represents a fermented ginseng berry Ganoderma lucidum product of Comparative Example 4.
  • the ginseng berry extract obtained in Preparation Example 1 and the fermented ginseng berry product powders obtained in Example 1 and Comparative Examples 1 to 4 were suspended in distilled water, and wrinkle alleviation effects at different concentrations (0.001%, 0.002%, 0.005%, 0.01%) were evaluated.
  • human dermal fibroblasts were irradiated with UVA at an energy of 5 J/cm 2 in a UV chamber.
  • Conditions of the dose of ultraviolet radiation and culture time which maximize expression of matrix metalloproteinase (MMP-1) in fibroblasts were established through preliminary testing.
  • a negative control group was wrapped with a foil and exposed to UVA for the same time.
  • UVA dose was measured using a UV radiometer.
  • the cells during irradiation of UVA were cultured in the medium previously used. After UVA irradiation, the cells were cultured in a fresh medium containing samples for 24 hours and 96-well plates were coated with the medium.
  • the cells were treated with primary antibodies ⁇ MMP-1 (Ab-5) monoclonal antibody, MMP-1 (Ab-3) monoclonal antibody ⁇ and reaction was conducted at 37° C. for 60 minutes.
  • the cells were reacted with anti-mouse IgG (whole mouse, alkaline phosphatase conjugated) as a secondary antibody for about 60 minutes, and were then reacted with an alkaline phosphatase substrate solution (1 mg/mL ⁇ -nitrophenyl phosphate in diethanolamine buffer solution) at room temperature for 30 minutes, and absorbance at 405 nm was measured using a microplate reader.
  • a control group was not treated with the sample.
  • the fermented ginseng berry products of the present invention (Example 1 and Comparative Examples 1 to 4) exhibited superior MMP-1 expression inhibitory capacity to the ginseng berry extract (Preparation Example 1).
  • the fermented Pleurotus ferulae product (Example 1) exhibited the highest MMP-1 exhibition inhibitory capacity.
  • MMP-1 exhibition inhibitory capacity increases, skin wrinkle alleviation effect increases.
  • the fermented ginseng berry Pleurotus ferulae product exhibited superior skin wrinkle alleviation effect to the ginseng berry extract.
  • ‘a’ represents a ginseng berry extract of Preparation Example 1
  • ‘b’ represents a fermented ginseng berry Pleurotus ferulae product of Example 1
  • ‘c’ represents a fermented ginseng berry Tricholoma matsutake product of Comparative Example 1
  • ‘d’ represents a fermented ginseng berry Sarcodon aspratus product of Comparative Example 2
  • ‘e’ represents a fermented ginseng berry Phellinus linteus product of Comparative Example 3
  • ‘f’ represents a fermented ginseng berry Ganoderma lucidum product of Comparative Example 4.
  • Tyrosinase (EC 1. 14. 18. 1) as an enzyme was dissolved in a 0.05M phosphate buffer (pH 6.5) such that a concentration of 1,100 unit/ml (final 55 unit) was obtained, L-tyrosine (C 9 H 11 NO 3 , 181.19) as a substrate was dissolved in a 0.1M phosphate buffer (pH 6.5) such that a concentration of 1.5 mM (final 225 uM) was obtained, the resulting solutions were each diluted with 0.1M sodium phosphate buffer (pH 6.5), and reacted (Table 3) and further reacted at 37° C.
  • the fermented ginseng berry Pleurotus ferulae product exhibited superior tyrosinase inhibition effect (whitening effect) to the ginseng berry extract.
  • Tyrosinase (EC 1.14.18.1) is an important enzyme which performs multiple enzymatic functions in the melanosynthetic pathway. In general, as tyrosinase inhibitory capacity increases, effects associated with skin whitening and propagation inhibition of melanoma formed by malignant alteration of melanine cells increase.
  • ‘a’ represents a ginseng berry extract of Preparation Example 1
  • ‘b’ represents a fermented ginseng berry Pleurotus ferulae product of Example 1
  • ‘c’ represents a fermented ginseng berry Tricholoma matsutake product of Comparative Example 1
  • ‘d’ represents a fermented ginseng berry Sarcodon aspratus product of Comparative Example 2
  • ‘e’ represents a fermented ginseng berry Phellinus linteus product of Comparative Example 3
  • ‘f’ represents a fermented ginseng berry Ganoderma lucidum product of Comparative Example 4.
  • a cosmetic composition containing 30.0% by weight of the fermented ginseng berry Pleurotus ferulae product obtained in Example 1 was prepared under the composition conditions shown in the following Table 4 and was referred to as “Formulation Example 1”, and a cosmetic composition not containing the fermented ginseng berry Pleurotus ferulae product was prepared and referred to as “Formulation Comparative Example 1”.
  • Formulation Example 1 the cosmetic containing the fermented ginseng berry Pleurotus ferulae product according to the present invention exhibited a considerably high skin conductance increase, as compared to Formulation Comparative Example 1.
  • skin conductance increases in proportion to skin water content the cosmetic containing the fermented ginseng berry Pleurotus ferulae product according to the present invention maintained high skin water content, as compared to a cosmetic not containing the fermented ginseng berry Pleurotus ferulae product (not shown).
  • transepidemal water losses (TEWL) of humans to whom the cosmetic of Formulation Example 1 and the cosmetic of Formulation Comparative Example 1 were applied were measured using a Tewameter (TM300, Courage and Khazaka Electronic Co., Germany) and were then compared.
  • the samples were applied to the right and left sides of faces of 40 women in their 20s to 40s, transepidemal water loss of an area spaced 3 cm apart to the right and 1 cm apart downward from the eye site was measured three times using Tewameter (TM300, Courage and Khazaka Electronic Co., Germany) before application and 1 hour, 2 hours, 4 hours and 6 hours after application and an average of three transepidemal water loss values was calculated.
  • transepidemal water loss on the facial skin of subjects to whom the cream prepared in Formulation Example 1 was applied was decreased.
  • the following experiment was performed in order to confirm the effect of the cosmetic containing the fermented ginseng berry Pleurotus ferulae product according to the present invention on atopy alleviation.
  • the cream of Formulation Example 1 and the cream of Formulation Comparative Example 1 were continuously applied to the atopy skin site of 10 subjects and the remaining 10 subjects, respectively, twice per day over four weeks.
  • the fermented ginseng berry Pleurotus ferulae product according to the present invention exhibits considerably superior anti-oxidation, anti-inflammation, collagen synthesis facilitation, skin wrinkle care, whitening, moisturizing, skin barrier improvement and atopy alleviation effects, as compared to a ginseng berry extract.
  • the present invention also provides a cosmetic composition having superior functionality.

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