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KR101861043B1 - Composition for Improving Skin Wrinkles Using an Extract of Penthorum chinense - Google Patents

Composition for Improving Skin Wrinkles Using an Extract of Penthorum chinense Download PDF

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KR101861043B1
KR101861043B1 KR1020170106611A KR20170106611A KR101861043B1 KR 101861043 B1 KR101861043 B1 KR 101861043B1 KR 1020170106611 A KR1020170106611 A KR 1020170106611A KR 20170106611 A KR20170106611 A KR 20170106611A KR 101861043 B1 KR101861043 B1 KR 101861043B1
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composition
extract
skin
food
present
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Korean (ko)
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김민진
김상철
신수영
안은정
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국립낙동강생물자원관
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/56Materials from animals other than mammals
    • A61K35/618Molluscs, e.g. fresh-water molluscs, oysters, clams, squids, octopus, cuttlefish, snails or slugs
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/98Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
    • A61K8/987Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of species other than mammals or birds
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • A61Q19/08Anti-ageing preparations
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00Function of food ingredients
    • A23V2200/30Foods, ingredients or supplements having a functional effect on health
    • A23V2200/318Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2250/00Food ingredients
    • A23V2250/20Natural extracts
    • A23V2250/204Animal extracts
    • A23V2250/2042Marine animal, fish extracts

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
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  • Gerontology & Geriatric Medicine (AREA)
  • Dermatology (AREA)
  • Marine Sciences & Fisheries (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Cosmetics (AREA)

Abstract

The present invention relates to a method for inhibiting the activity of elastase and inhibiting the expression of collagenase (MMP-1), and a method of inhibiting the expression of collagenase (MMP-1) by using a human fibroblast cell, CCD-986SK cell, A composition for improving wrinkles is disclosed.

Description

Technical Field [0001] The present invention relates to a composition for improving skin wrinkles,

The present invention relates to a pest The present invention relates to a composition for improving skin wrinkles.

The skin occupies about 16% of the human body and is directly in contact with the external environment and plays an important role in protecting the human body from external harmful factors such as temperature, humidity and ultraviolet rays. The skin consists of epidermis, dermis and subcutaneous tissue under keratin skin. The epidermis is composed of keratinocytes mainly composed of keratinocytes, a thin protective layer composed of melanocytes producing melanin and secreted melanocytes, immune cells such as langerhans cells and Merkel's copuscles To prevent external stimuli and pathogens from infestation, to control body temperature, to maintain moisture and lipid components. The dermis is a connective tissue beneath the epidermis, mostly composed of a macromolecular network called the extracellular matrix. This extracellular epilepsy is made up of fibroblasts and consists of fibrous proteins such as collagen, elastin and polysaccharides such as hyaluronic acid.

As the age increases, skin cells are damaged due to various pollutants, strong ultraviolet rays, stress and nutritional deficiency, and cell proliferation is not smooth, resulting in wrinkles, elastic loss and keratinization (Gilchrest BA, J. Am. Acad. Dermatol, 21: 610, 1989). Such skin aging is divided into endogenous aging and extrinsic aging. Endogenous aging is a natural aging phenomenon caused by age-related decline in the physiological function of the skin, resulting in reduced elasticity, rough skin, deep wrinkles, and pigmentation. Exogenous aging refers to aging caused by external factors such as ultraviolet light, reactive oxygen, and stress. When the aging of the skin progresses due to various factors as described above, reduction of keratinocyte and fibroblast cleavage, reduction of hyaluronic acid, decrease of collagen synthesis, increase of collagenase (MMP-1 etc.) production, increase of signal transmission of melanin production Resulting in reduction of moisturizing, generation of wrinkles and reduction of skin elasticity.

Collagen is the most important protein in maintaining skin hydration and maintaining skin flexibility and elasticity, accounting for 90% of the dermis (Journal of the American Academy of Dermatology, 2001, 17 (4): 610-613). Collagen is synthesized by fibroblasts and degraded by collagenase. When exposed to the sun or ultraviolet rays, the action of fibroblasts and cell number decrease, collagen synthesis decreases and collagenase activity, which increases collagenase activity, increases skin moisture loss and reduces skin elasticity and elasticity.

Therefore, a substance capable of inhibiting the expression or activity of the elastase, which degrades the collagenase or elastin, may have an effect of improving the wrinkles of the skin.

Currently, the effects of retinoic acid and retinol (J Invest Dermatol 98: 248-260, 1992; Elaine S et al., J Invest Dermatol 96: 975-978, 1991) on retinoic acid and retinol Ginsenoside Rg3 (Kim SW et al., J Soc Cosmet Scientists Korea 30: 221-225, 2004), which is extracted from ginseng, has been proved to be effective in the treatment of dihydroepiandrosterone (Shin MH et al., J Invest Dermatol 124: 315-323, , Wrinkle improvement of a variety of substances such as flavonoids (Francesco B et al., Int J Pharm 145: 87-94, 1996), Sorbus alba extract (Korean Patent No. 101008833), Yeast Extract (Korean Patent No. 100825450) Skin protection effects were discovered. However, some substances have limited use because they cause skin irritation and have safety problems. Accordingly, the search for a substance having an excellent wrinkle-improving activity effect among natural products having relatively high safety has been actively researched.

Octopus Bridge ( Penthorum chinense ) is a perennial herb that is distributed in Korea, China, and Japan. It is an aquatic plant that lives in wetlands such as nails and ditches. The shape of the fruit attached to the upper part of the stem is called 測 測?? Because it resembles the leg of the octopus. All parts of flowers, leaves and stems of Octopus legs have been used as medicinal materials and have been used for promoting blood circulation and used for amenorrhea, edema, uterine bleeding, cataract, bruise, etc. (Korean Medical Dictionary, Korean Medical Dictionary Compilation Committee, 2001). However, there is a lack of scientific evidence on the efficacy of octopus legs. In particular, no research has been conducted on the effect of wrinkles on skin cells.

The present invention discloses octopus leg extract having elastase inhibitory activity and inhibiting collagenase expression.

It is an object of the present invention to provide a composition for improving skin wrinkles using an Octopus legs extract.

Other and further objects of the present invention will be described below.

The present invention is based on the finding that the octopus leg extract inhibits the activity of elastase and inhibits the expression of collagenase (MMP-1) and inhibits the expression of human fibroblast cell, CCD-986SK Cells were found to show no specific cytotoxicity.

In view of the foregoing, the composition for improving skin wrinkles of the present invention is characterized by containing octopus leg extract as an active ingredient.

In the present specification, the term "octopus legs extract" means water, a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.) ), Methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, N, N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), 1,3-butylene glycol, An extract obtained by leaching using these mixed solvents, an extract obtained by using a supercritical extraction solvent such as carbon dioxide or pentane, or a fraction obtained by fractionating the extract, and the extraction method is a method in which the polarity, extraction degree, It is possible to apply any method such as cold rolling, refluxing, heating, ultrasonic irradiation, supercritical extraction, etc. In the case of the fractionated extract, a fraction obtained by suspending the extract in a specific solvent and mixing and leaving with a solvent having a different polarity, the crude extract is adsorbed on a column packed with silica gel, and then a hydrophobic solvent, a hydrophilic solvent, Quot; means fractions obtained as a mobile phase. Also, the meaning of the extract includes a concentrated liquid extract or a solid extract in which the extraction solvent is removed by a method such as freeze drying, vacuum drying, hot air drying, spray drying and the like. Preferably means an extract obtained by using water, a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.) or a mixed solvent thereof as an extraction solvent.

In the present specification, the term " active ingredient "alone means an ingredient which exhibits the desired activity or which can exhibit activity together with a carrier which is not itself active.

The composition of the present invention may contain, in addition to the active ingredient, an addition of similar activities such as enhancement or enhancement of skin wrinkle reducing effect or skin irritation inhibiting activity, skin protecting activity (skin whitening, skin damage suppression by ultraviolet rays, May further comprise any compound or natural extract known to be safe in the art and known to have a corresponding activity in order to improve the dosage, ingestion, and ease of use. These compounds or extracts include the national pharmacopoeia (Korean pharmacopoeia), the national health functional food circulation in Korea (in Korea, the health functional food standard and specification), the functional cosmetics circulation in each country (in Korea, Compounds or extracts listed in the official documents such as "Functional Cosmetics Standards and Test Methods"), compounds or extracts approved by the respective countries in accordance with the laws of the respective countries (in Korea, "Pharmaceutical Affairs Law") regulating the manufacture and sale of pharmaceuticals In accordance with the laws of each country that regulate the manufacture and sale of compounds or extracts and functional cosmetics that are recognized as functioning in accordance with the laws of the respective countries ("Act on Health Functional Foods" in Korea) that regulate the manufacture and sale of functional foods &Quot;).≪ / RTI > Examples of skin whitening ingredients include arbutin, niacinamide, ascorbyl glucoside, alpha-bisabolol and oil soluble licorice (Glycyrrhiza) extract. Examples of the skin wrinkle improving agent include retinol, retinyl palmitate , Adenosine, polyethoxylated amide, and the like. Examples of the ultraviolet protecting component include drometrizol, drometrizol trisiloxane, dipalayyltriolate, Dimethicyldecibenzalmalonate, diethylhexylbutadimido Triazone, and pine bark extract, phosphatidylserine, finger root extract powder, and complex extracts of red ginseng and mountain juice. Examples of the moisturizing component include AP collagen enzyme digesting peptide, collactive collagen peptide, N-acetyl glucosamine, konjac potato extract, dandelion extract, rice bran extract, corn germ extract, low molecular weight collagen peptide, ground extract powder, phosphatidylserine, hyaluronic acid Examples of functional ingredients for improving 'irritable skin condition' include L. sakei Probio 65, gamma linolenic acid-containing oil, L. plantarum CJLP133, and probiotic ATP. Such compounds or natural extracts may be included in the compositions of the present invention in combination with one or more of their active ingredients.

The composition of the present invention may contain any amount (effective amount) of the active ingredient as long as it can exhibit wrinkle-improving activity and the like which is intended to be treated depending on the purpose of use, formulation, compounding purpose and the like. And will be determined within the range of from 0.001% by weight to 15% by weight, based on the total weight of the composition. Herein, "effective amount" means a dermatologic / pharmacological effect, such as a wrinkle-reducing effect, when the composition of the present invention is administered to a mammal, preferably a human, Refers to the amount of active ingredient contained in the composition of the present invention. Such effective amounts can be determined experimentally within the ordinary skill of those skilled in the art.

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In a specific embodiment, the composition of the present invention can be identified as a food composition.

The food composition of the present invention can be produced in any form and can be used in various forms such as beverages such as tea, juice, carbonated drink, ionic drink, processed oil such as milk and yogurt, gum, rice cake, Korean confectionery, A food, a health food, a food, a tablet, a capsule, a ring, a granule, a liquid, a powder, a slice, a paste, a syrup, a gel, a jelly and a bar.

In addition, the food composition of the present invention may be classified into any product category as long as it meets the laws and regulations on the time of manufacture and distribution in the legal and functional category. For example, it is a health functional food according to the 「Health Functional Food Act」 in Korea, or a food functional food according to the Korean Food Sanitation Law (Food Standards and Specifications) , Special-purpose food, and the like.

The food composition of the present invention may contain food additives in addition to the active ingredients thereof. Food additives are generally understood to be substances that are added to foods and mixed or infiltrated into food in the manufacture, processing or preservation of food, and their safety must be ensured since they are ingested daily with food and for long periods of time. In food additives according to the laws of the respective countries ("Food Sanitation Act" in Korea) regulating the manufacture and distribution of food, food additives with safety are specified in terms of ingredient or function. In the Food Additives Code of Korea (Food Additives Standards and Standards), the food additives are classified into chemical compounds, natural additives and mixed preparations in terms of ingredients. Such food additives are classified into sweeteners, flavors Preservatives, emulsifiers, acidulants, and thickeners.

The sweetener is used for imparting a sweet taste suitable for foods, and both natural and synthetic sweeteners can be used in the composition of the present invention. Preferably, natural sweeteners are used. Examples of natural sweeteners include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose and maltose.

Flavors may be used to enhance taste or flavor, both natural and synthetic. Preferably, a natural one is used. When using natural ones, the purpose of nutritional fortification can be performed in addition to the flavor. Examples of natural flavoring agents include those obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, and the like, or those obtained from green tea leaves, Asiatica, Daegu, Cinnamon, Chrysanthemum leaves and Jasmine. Also, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, banks and the like can be used. The natural flavoring agent may be a liquid concentrate or a solid form of extract. Synthetic flavors may be used depending on the case, and synthetic flavors such as esters, alcohols, aldehydes, terpenes and the like may be used.

As the preservative, calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate and EDTA (ethylenediaminetetraacetic acid) can be used. As the emulsifier, acacia gum, carboxymethyl cellulose, Pectin and the like. As the acidulant, math, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid and the like can be used. The acidulant may be added so that the food composition has a proper acidity for the purpose of inhibiting the growth of microorganisms other than the purpose of enhancing the taste.

Examples of the thickening agent include suspending agents, sedimentation agents, gel-forming agents, bulking agents and the like.

The food composition of the present invention may contain physiologically active substances or minerals which are known in the art and which are stable as a food additive in addition to the above-mentioned food additives in order to supplement and supplement functional and nutritional properties.

Examples of such physiologically active substances include catechins contained in green tea and the like, vitamins such as vitamin B1, vitamin C, vitamin E and vitamin B12, tocopherol, dibenzoyl thiamine, etc. Examples of minerals include calcium preparations such as calcium citrate, magnesium stearate , Iron preparations such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, zinc and the like.

The food composition of the present invention may contain an appropriate amount of the above-mentioned food additives according to the product type so as to achieve the purpose of addition thereof.

With regard to other food additives that may be included in the food composition of the present invention, reference may be made to the Food Code of the respective countries or the Food Additives Code.

In another specific embodiment, the composition of the present invention can be identified as a pharmaceutical composition.

The pharmaceutical composition of the present invention may be prepared into oral formulations or parenteral formulations according to the route of administration by conventional methods known in the art, including pharmaceutically acceptable carriers in addition to the active ingredient. Where the route of administration may be any suitable route including local routes, oral routes, intravenous routes, intramuscular routes, and direct absorption through mucosal tissues, and combinations of two or more routes may be used. An example of a combination of two or more routes is a combination of two or more formulations of the drug according to the route of administration, for example, one drug is administered intravenously and another drug is administered via a local route.

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration and formulation, and specific reference may be made to the pharmacopoeia of each country, including the "Korean Pharmacopoeia ".

When the pharmaceutical composition of the present invention is prepared into an oral formulation, it may be formulated into powder, granules, tablets, pills, sugar tablets, capsules, solutions, gels, syrups, suspensions, wafers And the like. Examples of suitable carriers include starches such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol and xylitol, corn starch, potato starch and wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Hydroxypropylmethylcellulose and the like; polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol Serol, and the like. In case of formulation, suitable binders, lubricants, disintegrants, coloring agents, diluents and the like may be included as needed. Examples of suitable binders include starch, magnesium aluminum silicate, starch pellets, gelatin, methyl cellulose, sodium carboxymethyl cellulose, polyvinyl pyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax and the like. Examples of the disintegrating agent include starch, methylcellulose, magnesium stearate, magnesium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, Agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt, and the like. Examples of the diluent include lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine and the like.

When the pharmaceutical composition of the present invention is prepared into a parenteral dosage form, it may be formulated in the form of an injection, transdermal drug delivery, nasal aspirate and suppository together with a suitable carrier according to methods known in the art. As the carrier suitable for injection preparation, aqueous isotonic solutions or suspensions may be used. Specifically, PBS (phosphate buffered saline) containing triethanolamine, sterile water for injection, and isotonic solution such as 5% dextrose may be used . When formulated with a transdermal preparation, it can be formulated in the form of ointments, creams, lotions, gels, external liquids, pastes, liniments, and air-lozenges. Nasal inhalers may be formulated in the form of aerosol sprays using suitable propellants such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc., and when formulated as a suppository, witepsol, tween 61, polyethylene glycols, cacao butter, laurin, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, and sorbitan fatty acid esters.

The formulation of pharmaceutical compositions is well known in the art and can be found, for example, in Remington ' s Pharmaceutical Sciences (19th ed., 1995). This document is considered part of this specification.

The preferred dosage of the pharmaceutical composition of the present invention is 0.001 mg / kg to 10 g / kg per day, preferably 0.001 mg / kg to 1 g / day, depending on the patient's condition, body weight, sex, age, / kg < / RTI > The administration can be carried out once or several times a day. Such dosages should in no way be construed as limiting the scope of the invention.

In another specific embodiment, the composition of the present invention can be identified as a cosmetic composition.

Even when the composition of the present invention is identified as a cosmetic composition, the cosmetic composition may be classified into any product category according to the use of the cosmetic composition. Specifically, the cosmetic composition may contain functional cosmetic products having applications such as improvement of skin troubles and improvement of atopic dermatitis, General cosmetics, and the like. Specific examples of the product form include a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing oil, powder foundation, emulsion foundation, wax Foundation, spray, and the like. In a specific product form, it may be a form of flexible lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.

The cosmetic composition of the present invention may contain, in addition to its active ingredient, conventional additives such as stabilizers, solubilizing agents, surfactants, vitamins, colorants and antioxidants, and carriers commonly used in cosmetic compositions.

When the formulation of the present invention is a paste, a cream or a gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent is used as a carrier component. Specifically, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid esters of sorbitan, and the like.

When the formulation of the present invention is a suspension, a carrier, such as water, a liquid diluent such as ethanol or propylene glycol, a suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component is selected from aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters.

The cosmetic composition of the present invention can be produced according to a method for producing a cosmetic composition which is usually carried out in the art, except that it contains the effective ingredient showing skin wrinkle improving activity.

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INDUSTRIAL APPLICABILITY As described above, the present invention can provide a composition for improving skin wrinkles using an octopus leg extract.

The composition for improving skin wrinkles of the present invention can be produced into foods, cosmetics, medicines and the like.

1 is an experimental result on the elastase inhibitory activity of octopus legs extract
Fig. 2 shows the cytotoxicity test results of octopus legum extract.
Fig. 3 shows experimental results on the inhibitory activity of collagenase (MMP-1) expression of Octopus legum extract.

Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these examples and experimental examples.

< Example > Experiment of skin wrinkle inhibition activity of Octopus legum extract

I. Materials and Methods

Preparation of sample

The octopus legs used in this experiment were sampled and identified in the vicinity of Gyeonggi Province and shaded. The shrubs were shredded and used as experimental material.

Extraction of sample

The extracts were collected by filtration (Whatman No.2) three times at room temperature for 24 hours with 70% ethanol at 10 times the weight of octopus leg or seed dry powder, concentrated by rotary vacuum evaporator (EYELA. Japan) And stored at -20 ° C until used.

Measurement of Elastase Inhibitory Activity

40 μl of each test solution was added to 40 μl of a solution of proporcine pancreas elastase (2.5 U / ml) dissolved in 50 mM Tris-HCl buffer (pH 8.6) 80 μl of N-succinyl- (L-Ala) 3-p-nitroanilide (0.5 mg / ml) was added to the reaction mixture at 37 ° C for 30 minutes. The amount of pnitroanilide produced from the substrate was measured at 410 nm. Elastase inhibitory activity was expressed as the absorbance reduction rate of the sample solution added and no added sample solution.

Inhibition rate (%) = 1- (absorbance of sample addition group / absorbance of sample addition group) x 100

Cell culture

The CCD-986SK used in this experiment was purchased from ATCC (American Type Culture Collection, USA) and used for the experiment. The medium was supplemented with Dulbecco's modified Eagle's medium (DMEM) (GIBCO Inc, NY, USA) containing 100 units / ml penicillin-streptomycin (GIBCO Inc, NY, USA) and 10% fetal bovine serum (FBS; Gibco, Grand Island, USA) USA) medium at 37 ° C in a 5% CO 2 incubator, and subculture was performed once every 3-4 days.

Cytotoxicity test (MTT assay)

MTT analysis was performed to determine the effect of the sample on cell growth. The living cells, which are rich in metabolism, reduce the water-soluble yellow 3- (4,5-dimethylthiaxo-2-yl) -2,5-diphenyl tetrazolium bromide by the dehydrogenase action of intracellular mitochondria, Forms soluble formazan.

The CCD-986SK cells were cultured in DMEM medium supplemented with 10% FBS at a concentration of 5.0 × 10 4 cells / well in a 96-well plate for 18 hours and then treated for 24 hours. Then, 50 μl of MTT solution was added and reacted for 4 hours. After the culture medium was completely removed, 200 μl of dimethylsulfoxide was added to completely dissolve the precipitate, and the absorbance at 540 nm was measured using a microplate reader. The average absorbance value of each sample group was determined and compared with the absorbance value of the control group, the cell growth rate was evaluated.

Measurement of MMP-1 expression inhibitory activity

Cells were inoculated into 24-well plates at a density of 5.0 × 10 4 cells / well using DMEM-subcultured fibroblasts and incubated for 18 h. The sample solution was added to each concentration and cultured for 48 h in serum-free medium. To increase the activity of MMP-1, TNF-α was added at a concentration of 50 ng / mL. Then, the amount of MMP-1 expressed in the supernatant obtained by centrifuging the cell culture medium was measured. All samples were stored frozen until quantification. The expression level of MMP-1 was quantitated using the pro-Matrix Metalloproteinase-1 kit (R & D system). The r2 value of the standard curve for the standard was 0.99 or more.

Ⅱ. Results and Discussion

Measurement of Elastase Inhibitory Activity

The elastase present in the neutrophil granulocyte of human body is known as the main enzyme of wrinkle formation by decomposing elastin which is insoluble elastic fiber protein of animal connective tissue and breaking the network structure of dermal tissue of skin. The effect of top seed and extract of octopus on elastase activity was examined.

As shown in Fig. 1, the results of confirming elastase inhibitory activity of the octopus legs and the seed extract at a concentration range of 400 / / mL or less showed that the octopus legumes and seed extracts exhibited elastase inhibitory activity in a concentration-dependent manner. Especially, in the case of Octopus legume seed extract, excellent elastase inhibitory effect similar to Oleanolic acid, which is known as a wrinkle remover at a concentration of 100 ㎍ / mL, was confirmed (FIG. 1).

Cytotoxicity Assessment

Human fibroblast cells, CCD-986SK cells, were cultured in a 24-well plate at 5.0 × 10 4 cells / well for 24 hours, and then treated with octopus legume seed and seed extracts at a concentration of 200 μg / mL or less for 24 hours MTT assay was used to confirm cell viability.

As shown in FIG. 2, the octopus legumes and seed extracts showed cell viability of 90-100% in the concentration range of 100 μg / mL and 200 μg / mL, respectively, indicating that they did not show cytotoxicity. Therefore, the following experiments were carried out at concentrations that did not show cytotoxicity for each of the extracts (Fig. 2).

Measurement of MMP-1 expression inhibitory activity

Collagen is one of the major components of skin connective tissue. Therefore, decomposition of collagen directly affects the elasticity of connective tissues and wrinkles and defects of the skin. Among the dozens of MMPs produced in the body, MMP-1 is a collagen-specific proteinase that inhibits the activity of MMP-1 and protects collagen to maintain the elasticity of the skin tissue and produce wrinkles It is known to be preventable. Therefore, we examined the inhibitory effect of octopus legum extract on MMP-1 activity.

As shown in Fig. 3, the octopus legume extract (12.5, 25, 50, and 100 μg / mL) and seed extracts (25, 50, 100, and 200 μg / mL) / mL) and cultured for 48 hours. The expression level of MMP-1 was measured using an ELISA kit.

It was confirmed that the expression of MMP-1 was induced by TNF-α, and that the seed extract inhibited MMP-1 expression in a concentration-dependent manner than that of Octopus leg (Fig. 3).

Claims (6)

A composition for improving skin wrinkles comprising an extract of Penthorum chinense as an active ingredient.
The method according to claim 1,
Wherein the octopus legs extract is obtained by extracting octopus legs outpox, leaf, stem, root, ground part, ground part or seed with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.
The method according to claim 1,
Wherein the octopus legs extract is obtained by extracting octopus legs outpowder or seeds with methanol.
4. The method according to any one of claims 1 to 3,
The composition for improving skin wrinkles is characterized in that the composition is a cosmetic composition.
4. The method according to any one of claims 1 to 3,
The composition for improving skin wrinkles is characterized in that the composition is a food composition.
4. The method according to any one of claims 1 to 3,
The composition for improving skin wrinkles is characterized by being a pharmaceutical composition.




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KR101993847B1 (en) * 2018-11-28 2019-06-27 (주)그린솔루션스 Cosmetic composition for improving skin wrinkles or skin moisturizing with native plants in DMZ
KR102044891B1 (en) * 2019-05-17 2019-11-14 (주)그린솔루션스 Cosmetic composition for improving skin wrinkles or skin moisturizing with native plants in DMZ
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KR102221408B1 (en) 2020-10-27 2021-03-02 한국콜마주식회사 Cosmetic composition comprising extracts of rubus coreanus miq. and extracts of penthorum chinense pursh for moisturizing the skin
KR102406286B1 (en) * 2021-12-23 2022-06-13 국립낙동강생물자원관 Antibacterial composition using extract of Penthorum chinense

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KR20200052029A (en) * 2018-11-06 2020-05-14 재단법인 경기도경제과학진흥원 cosmetic composition for promoting skin regeneration comprising the extract of Penthorun chinense Pursh
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KR101993847B1 (en) * 2018-11-28 2019-06-27 (주)그린솔루션스 Cosmetic composition for improving skin wrinkles or skin moisturizing with native plants in DMZ
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KR102221408B1 (en) 2020-10-27 2021-03-02 한국콜마주식회사 Cosmetic composition comprising extracts of rubus coreanus miq. and extracts of penthorum chinense pursh for moisturizing the skin
KR102406286B1 (en) * 2021-12-23 2022-06-13 국립낙동강생물자원관 Antibacterial composition using extract of Penthorum chinense

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