KR102305237B1 - Compositions for Improving Skin Wrinkles and Skin Whitening Using an Extract of Persicaria filiforme - Google Patents

Abstract

The present invention discloses a composition for skin wrinkle improvement and skin whitening using an extract showing elastase inhibitory activity and also showing tyrosinase inhibitory activity.

Description

Compositions for Improving Skin Wrinkles and Skin Whitening Using an Extract of Persicaria filiforme}

The present invention relates to a composition for skin wrinkle improvement and skin whitening using an extract of Persicaria filiforme.

The skin occupies about 16% of the human body and is in direct contact with the external environment, so it plays an important role in protecting the human body from external harmful factors such as temperature, humidity and ultraviolet rays. The skin is composed of the epidermis, dermis, and subcutaneous tissue under the keratin. The epidermis is a thin protective layer composed of keratinocytes, which are mainly made of keratin, melanocytes that produce and secrete melanin, immune cells such as Langerhans cells, and Merkel's copuscles. It prevents external stimuli and pathogens from entering, regulates body temperature, and maintains moisture and lipid components. The dermis is a connective tissue beneath the epidermis and is mostly composed of a network of macromolecules called extracellular matrix. This extracellular matrix is made from fibroblasts and consists of fibrous proteins such as collagen and elastin, and polysaccharides such as hyaluronic acid.

As we age, skin cells are damaged due to various contaminants, strong ultraviolet rays, stress, and nutritional deficiencies. Gilchrest BA, J. Am. Acad. Dermatol, 21, 610. 1989).

Collagen is the most important protein for maintaining skin moisture and maintaining skin flexibility and elasticity, and it occupies 90% of the dermal layer (J Am Acad Dermatol, 17(4):610-613. 2001) and gives skin strength and tension. It serves to protect the skin from external stimuli. In addition, elastin in the dermal layer accounts for about 2%, which is less than collagen, but plays a role in supporting collagen fibers. It has been reported in many recent studies that the deficiency, aggregation, and loss of elastin fibers play an important role in the mechanism of skin wrinkle formation. (Weihermann AC et al., Int J Cosmet Sci. 2017 Jun;39(3):241-247; Mora Huertas AC et al., Biochimie. 2016 Sep-Oct;128-129:163-73).

Collagen and elastin are synthesized in fibroblasts and are degraded by collagenase or elastase. As people age or are exposed to UV rays, the action and cell number of fibroblasts decreases, the amount of collagen or elastin synthesized decreases, and the action of collagenase or elastase that decomposes them increases. Once the elasticity of the skin is reduced by collagen and elastin, it is very difficult to recover. Accordingly, research for improving skin wrinkles by inhibiting elastase activity, which increases with age, is being actively conducted.

Currently, as a substance that inhibits skin wrinkle formation, retinoic acid and retinol (Simon C et al., J Invest Dermatol 98: 248-260. 1992; Elaine S et al., J Invest Dermatol 96: 975-978. 1991) Efficacy has been proven, dehydroepiandrosterone (Shin MH et al., J Invest Dermatol 124: 315-323.2005), ginsenoside Rg3 extracted from ginseng (Kim SW et al., J Soc Cosmet Scientists Korea 30: 221-225.2004), flavonoids ( Francesco B et al., Int J Pharm 145: 87-94.1996), red bean extract (Korean Patent No. 101008833), Yeongyo extract (Korean Patent No. 100825450), etc. Wrinkle improvement and skin protection effects were explored. became However, some substances have low stability or cause skin irritation, so there is a problem with safety, so their use is limited. Accordingly, the search for substances with excellent anti-wrinkle activity, safety and stability is continuing.

It is known that melanin is synthesized in melanocytes, which are color cells present in the basal layer of the skin (epidermis), and metastasizes to surrounding keratinocytes to represent the color of human skin. When the skin is exposed to sunlight, a chemical reaction occurs in melanocytes, which are pigment-forming cells, and melanin is produced. The main role of melanin is to protect the inside of the skin by removing free radicals and preventing the penetration of UV rays by active oxygen generated in the skin (Brain Research Bulletin 10 (6): 847 851, 1983; Mutation Research 571 (1): 121-2, 2005; Nature. 22;445(7130):843-50. 2007). However, overexpression of melanin greatly affects not only skin health but also aesthetic aspects such as skin blackening, pigmentation, melasma, freckles, and skin cancer (Pigment Cell Res 10, 127-138. 1997; Vet Dermatol. 2003 Apr; 14(2) ):57-65.).

The starting material for making melanin is tyrosine, a type of amino acid normally present in the human body. Tyrosine is oxidized by an enzyme called tyrosinase in melanocytes and converted to dihydroxy phenylalanine (DOPA), which is further oxidized to produce dopaquinone (DOPA-Quinone). And when dopaquinone meets cysteine, cysteinyldopa is produced, and as a result, reddish-brown pheomelanin is produced. The produced melanin is then loaded onto melanosomes and delivered to keratinocytes (Pigment Cell Res.12(1):4-12.1999.; Cell Mol Biol (Noisy-le-grand).45(7):981). -90.1999;An Bras Dermatol.88(1):76-3.2013).

Accordingly, the basic mechanisms of drugs formulated in whitening cosmetics for the purpose of preventing or alleviating pigmentation are inhibition of tyrosinase action, inhibition of tyrosinase production, inhibition of melanin production mediator, reduction of existing melanin and inhibition of strong oxidation, and excretion of melanin. promotion, and UV protection. Representative substances that inhibit the activity of tyrosinase include arbutin, kojic acid, and the like. Substances that directly destroy melanin include hydroquinone and mercury, but the use of mercury is prohibited as a toxic substance. Whitening agents that reduce the produced melanin include vitamin C, glutathione, magnesium ascrobic acid phosphate, etc. (J Cosmet Sci.65(6):365-75. 2014; Int J Cosmet Sci.37 ( 6):567-73.2015;Yakugaku Zasshi.128(8):1203-7.2008;Int J Cosmet Sci.27(3):147-53.2005;Acta Derm Venereol.63(3):209- 14.1983;Skin Pharmacol Appl Skin Physiol.13(3-4):143-9.2000;J Cosmet Laser Ther.15(2):107-13.2013.).

As interest in whitening is growing, development of whitening cosmetics with superior effects other than well-known substances is being actively developed. have. Therefore, recently, studies on natural whitening agents that do not affect cells and reduce melanin synthesis are being actively conducted.

The present invention discloses the skin wrinkle improvement activity and skin whitening activity of the extract

It is an object of the present invention to provide a composition for skin wrinkle improvement and skin whitening using the extract

Other objects or specific objects of the present invention will be set forth below.

As confirmed in the Examples and Experimental Examples below, the present invention has been completed by confirming that the E. serrata extract exhibits elastase inhibitory activity and also tyrosinase inhibitory activity.

Considering the above, in one aspect, the present invention can be identified as a composition for skin wrinkle improvement comprising an oleander extract as an active ingredient. can figure out

On the other hand, as confirmed in the following Examples and Experimental Examples, the present inventors macrophages that play a central role in the inflammatory response (Ito T., et al., Curr Drug Traget Inflamm Allergy, 2(3):257-265, 2003) was treated with LPS (lipopolysaccharide) to induce an inflammatory response, and then, when the nitric oxide (NO) production level was evaluated by treating the sample, lichen extract by concentration, the nitric oxide (NO) production was significantly increased It was confirmed that it inhibited (IC ₅₀ = 45.29 μg/ml), and it was confirmed that it did not show any particular cytotoxicity even at the highest treatment concentration of 100 μg/ml.

Nitric oxide is synthesized from L-arginine by the action of nitric oxide synthase (NOS), and NOS exists in several isoforms. bNOS (brain NOS) present in the brain, nNOS (neuronal NOS) present in the nervous system, and eNOS (endothelial NOS) present in the vascular endothelial system are always expressed at a certain level in the body, and monoxide produced in small amounts by these Nitrogen (NO) plays an important role in maintaining homeostasis of the human body by performing various physiological reactions related to blood pressure regulation, neurotransmission, learning, and memory. On the other hand, iNOS (induced NOS) whose expression is induced by a certain stimulus produces excessive NO, and the nitric oxide produced excessively by iNOS reacts with superoxide to form peroxynitrite and It acts as a powerful oxidizing agent and by damaging cells, it is involved in various pathological processes including inflammation and cancer (Gupta SC et al., Exp Biol Med., 236:658-671, 2011; Riehemann et al., FEBS). Lett., 442:89-94, 1999; Stamler et al., Science, 258:1898-1902, 1992).

In view of the above, the present invention can provide a composition for inhibiting nitric oxide production comprising an extract of lichen in another aspect as an active ingredient. Such a composition of the present invention can be usefully used for inflammatory diseases caused by excessive production of nitric oxide. Such inflammatory diseases include asthma, allergic and non-allergic rhinitis, chronic and acute rhinitis, chronic and acute gastritis or enteritis, ulcerative gastritis, acute and chronic nephritis, acute and chronic hepatitis, chronic obstructive pulmonary disease, pulmonary fibrosis, hypersensitivity. Bowel syndrome, inflammatory pain, migraine, headache, back pain, fibromyalgia, myofascial disease, viral infection (such as hepatitis C infection), bacterial infection, fungal infection, burns, surgical or dental wounds, prostaglia Dean E excess syndrome, atherosclerosis, gout, arthritis, rheumatoid arthritis, ankylosing spondylitis, Hodgkin's disease, pancreatitis, conjunctivitis, iritis, scleritis, uveitis, dermatitis (including atopic dermatitis), eczema, multiple sclerosis, etc. have.

On the other hand, in the present specification, as used herein, the term "Essea extract" refers to extracts of extracts of sagebrush, leaves, stems, above-ground parts, roots, rhizomes, underground parts, seeds, or mixtures thereof with water and lower alcohols 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), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol Or an extract obtained by leaching using a mixed solvent thereof, an extract obtained using a supercritical extraction solvent such as carbon dioxide or pentane, or a fraction obtained by fractionating the extract, and the extraction method is the polarity of the active material, extraction degree, preservation In consideration of the degree, any method such as chilling, reflux, warming, ultrasonic radiation, or supercritical extraction may be applied. In the case of a fractionated extract, a fraction obtained by suspending the extract in a specific solvent and mixing and standing still with a solvent having a different polarity, and adsorbing the crude extract to a column filled with silica gel, etc. It is meant to include the fraction obtained as a mobile phase. In addition, the meaning of the extract includes a concentrated liquid extract or solid extract from which the extraction solvent is removed by freeze drying, vacuum drying, hot air drying, spray drying, or the like. Preferably, it refers to 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. More preferably, a fractional extract obtained by suspending a mixed solvent extract of water and ethanol in distilled water and fractionation with hexane, dichloromethane, ethyl acetate or butanol, or each fractional extract obtained by sequential fractionation using the fractionation solvents. . Here, "sequential fractionation" means that the fraction of the residual water layer after fractionation is used continuously after fractionation to fractionate with the solvents in the order listed above.

In addition, as used herein, the term "active ingredient" refers to a component capable of exhibiting the desired activity alone or in combination with a carrier that has no activity by itself.

In addition to the active ingredient, the composition of the present invention has a similar activity, such as skin wrinkle improvement effect or skin whitening effect, for increasing/reinforcing or for skin hypersensitivity reaction inhibition activity, skin protection activity (skin damage suppression due to ultraviolet rays, skin moisturizing, etc.) In order to enhance the convenience of administration, ingestion, and use through addition, any compound or natural extract known to have the activity and safety has already been verified in the art may be additionally included.

These compounds or extracts include pharmacopoeia of each country (“Korea Pharmacopoeia” in Korea), health functional food regulations of each country (“health functional food standards and specifications” announced by the Ministry of Food and Drug Safety in Korea), functional cosmetics regulations of each country (the Ministry of Food and Drug Safety notice in Korea). Compounds or extracts listed in the compendium such as "Standards and Test Methods for Functional Cosmetics," etc.) In accordance with the laws of each country governing the manufacture and sale of functional foods (the “Health Functional Food Act” in Korea), each country’s laws governing the manufacture and sale of compounds or extracts and functional cosmetics whose functionality has been recognized (the “Cosmetics Act” in Korea). '), compounds or extracts whose functionality is recognized are included.

Such ingredients include, for example, arbutin, niacinamide, ascorbyl glucoside, alpha-bisabolol, oil-soluble licorice, recognized as a skin lightening ingredient in the Cosmetics Code (Korean Ministry of Food and Drug Safety Notification, 「Standards and Specifications for Functional Cosmetics」) according to the Korean Cosmetics Act. Extract (Oil Soluble Licorice (Glycyrrhiza) Extract), retinol, retinyl palmitate, adenosine, polyethoxylated amide, etc. recognized as skin wrinkle improvement ingredients in the same Korean Cosmetics Competition, and UV protection in the same Korean cosmetic industry Complex such as dromethrazole, dromethrazole trisiloxane, digalloyl trioleate, dimethicodiethylbenzalmalonate, diethylhexylbutamidotriazone, pine bark extract, phosphatidylserine, fingerroot Extracts powder, complex extracts of red ginseng and cornus oil, etc., and as functional ingredients 'improving sensitive skin condition' according to the Korean Health Functional Food Act, L. sakei Probio 65, oil containing gamma-linolenic acid, lactic acid bacteria derived from fruits and vegetables L. plantarum CJLP133, probiotic ATP, and the like. One or more of these compounds or natural extracts may be included in the composition of the present invention together with its active ingredient.

The composition of the present invention may contain any amount (effective amount) of the active ingredient as long as it can exhibit wrinkle improvement activity, skin whitening activity, NO production inhibitory activity, etc. intended for treatment according to use, formulation, compounding purpose, etc. , a typical effective amount will be determined within the range of 0.001% to 15% by weight, based on the total weight of the composition. Herein, the term "effective amount" refers to a wrinkle improvement effect, a skin whitening effect, an NO production inhibitory effect, etc. It refers to the amount of the active ingredient included in the composition of the present invention, which can exhibit one dermatological and medical effect. Such effective amounts can be determined empirically within the ordinary ability of one of ordinary skill in the art.

In a specific aspect, the composition of this invention can be grasped|ascertained as a food composition. When the composition for inhibiting NO production of the present invention is identified as a food composition, its use may be understood as 'for inhibiting inflammatory response'.

The food composition of the present invention may be prepared in any form, for example, beverages such as tea, juice, carbonated beverages, and ionic beverages, processed oils such as milk and yogurt, gums, rice cakes, Korean sweets, bread, confectionery, noodles, etc. Foods, tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, and health functional food preparations such as bars can be prepared.

In addition, the food composition of the present invention may have any product classification in terms of legal and functional classification as long as it conforms to the enforcement laws at the time of manufacture and distribution. For example, it is a health functional food according to the law of an excluded country that regulates the manufacture and sale of health functional food (the “Health Functional Food Act” in Korea), or a law of an excluded country that regulates the manufacture and sale of food (in Korea, it is the “Health Functional Food Act”) According to the Food Sanitation Act (“Food Sanitation Act”), confectionery, beans, soy milk, fermented beverages, and special purpose foods according to each food type in accordance with the Food Standards and Specifications Notice of the Ministry of Food and Drug Safety in Korea.

The food composition of the present invention may contain food additives in addition to the active ingredients thereof. Food additives can be generally understood as substances that are added to and mixed with or infiltrated into food in manufacturing, processing, or preserving food. Food additives with guaranteed safety are limited in terms of ingredients or functions in the Food Additives Ordinance in accordance with the laws of each country that regulates the manufacture and distribution of food (“Food Sanitation Act” in Korea). In the Korean Food Additives Code (“Food Additive Standards and Specifications” notified by the Ministry of Food and Drug Safety), food additives are classified into chemically synthetic products, natural additives, and mixed preparations in terms of ingredients. It is classified into an agent, a preservative, an emulsifier, an acidulant, and a thickener.

The sweetener is used to impart appropriate sweetness to food, and both natural and synthetic ones may be used in the composition of the present invention. Preferably, a natural sweetener is used. Examples of the natural sweetener include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

Flavoring agents may be used to improve taste or aroma, and both natural and synthetic ones may be used. Preferably, it is a case where a natural thing is used. In the case of using a natural one, the purpose of nutritional enhancement in addition to flavor may be concurrently used. The natural flavoring agent may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, or the like, or obtained from green tea leaves, horseradish leaves, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, ginkgo biloba, etc. can be used. The natural flavoring agent may be a liquid concentrate or a solid extract. Optionally, a synthetic flavoring agent may be used, and the synthetic flavoring agent may be an ester, an alcohol, an aldehyde, a terpene, or the like.

As a preservative, sodium calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. can be used, and as an emulsifier, acacia gum, carboxymethylcellulose, xanthan gum, Pectin etc. are mentioned, and acidulant, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, etc. can be used as an acidulant. Acidulant may be added so that the food composition has an appropriate acidity for the purpose of inhibiting the growth of microorganisms in addition to the purpose of enhancing the taste.

As the thickening agent, a suspending agent, a settling agent, a gel-forming agent, a bulking agent, and the like can be used.

The food composition of the present invention may contain, in addition to the food additives as described above, physiologically active substances or minerals known in the art for the purpose of supplementing and reinforcing functionality and nutrition and guaranteed stability as food additives.

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, dibenzoylthiamine, and the like. Magnesium preparations, such as iron preparations, such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, zinc, etc. are mentioned.

In the food composition of the present invention, the food additives as described above may be included in an appropriate amount to achieve the purpose of the addition according to the type of product.

In relation to other food additives that may be included in the food composition of the present invention, reference may be made to the national food regulations or food additives regulations.

The composition of the present invention may be identified as a pharmaceutical composition in another specific embodiment.

In particular, when the composition for skin whitening of the present invention is identified as a pharmaceutical composition, the target disease can be understood as skin hyperpigmentation, and specific examples of such hyperpigmentation include freckles, senile spots, eczema, melasma, brown or dark spots, and sunlight. Pigmentation, cyanic melasma, hyperpigmentation after use of drugs such as calcium antagonists, sequelae from sclerotherapy, gravidic chloasma, melasma in women taking oral contraceptives , post-inflammatory hyperpigmentation due to wounds or dermatitis, including abrasions and burns, phototoxic reactions, or other similar small fixed pigmented lesions, and the like.

The pharmaceutical composition of the present invention may be prepared as an oral dosage form or a parenteral dosage form according to the route of administration by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. Here, the route of administration may be any suitable route including topical route, oral route, intravenous route, intramuscular route, and direct absorption through mucosal tissue, and two or more routes may be used in combination. An example of the combination of two or more routes is a case in which two or more formulations of drugs according to the route of administration are combined. For example, one drug is first administered by an intravenous route and the other drug is secondarily administered by a local route.

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

When the pharmaceutical composition of the present invention is prepared as an oral dosage form, powders, granules, tablets, pills, dragees, capsules, liquids, gels, syrups, suspensions, wafers, together with a suitable carrier according to methods known in the art. It can be prepared in a formulation such as Examples of suitable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol, starches such as corn starch, potato starch, wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Cellulose such as hydroxypropylmethylcellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol, grease Serol etc. are mentioned. In the case of formulation activity, an appropriate binder, lubricant, disintegrant, colorant, diluent, etc. may be included as needed. Suitable binders include starch, magnesium aluminum silicate, starch ferrist, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax, and the like, and oleic acid as lubricant. sodium, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, its magnesium and calcium salts, polyethylene glycol, and the like. , agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt. 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 for parenteral use, it may be formulated in the form of injections, transdermal administrations, nasal inhalants and suppositories together with suitable carriers according to methods known in the art. When formulated as an injection, an aqueous isotonic solution or suspension may be used as a suitable carrier, and specifically, PBS (phosphate buffered saline) containing triethanolamine, sterile water for injection, or isotonic solution such as 5% dextrose may be used. . When formulated for transdermal administration, it can be formulated in the form of ointments, creams, lotions, gels, external solutions, pasta agents, liniment agents, air rolls, and the like. In the case of nasal inhalants, it can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, and the like. witepsol), tween 61, polyethylene glycols, cacao fat, laurin fat, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, sorbitan fatty acid esters, and the like can be used.

Specific formulations of pharmaceutical compositions are known in the art, and reference may be made to, for example, Remington's Pharmaceutical Sciences (19th ed., 1995). This document is considered a part of this specification.

A preferred dosage of the pharmaceutical composition of the present invention is in the range of 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 1 g, depending on the patient's condition, weight, sex, age, patient's severity, and administration route. It can be in the range /kg. Administration may be performed once or divided into several times a day. Such dosages should not be construed as limiting the scope of the invention in any respect.

In another specific embodiment, the composition of the present invention can be identified as a cosmetic composition. When the anti-inflammatory composition of the present invention is identified as a cosmetic composition, its use may be understood as alleviation of inflammatory skin irritation.

Even when the composition of the present invention is identified as a cosmetic composition, the cosmetic composition can be classified into any product in terms of its use or by law, and specifically functionalities with uses such as improvement of skin troubles, improvement of skin whitening, improvement of atopic dermatitis, etc. It may be cosmetics, non-functional general cosmetics, and the like. The product form may also take any product form, specifically solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax It can take the form of products such as foundation and spray. In the specific product form, it may be in the form of a flexible lotion, a nourishing lotion, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray, or a powder.

The cosmetic composition of the present invention may include components commonly used in cosmetic compositions in addition to the active ingredient, for example, conventional adjuvants such as stabilizers, solubilizers, surfactants, vitamins, pigments and fragrances, and carriers.

When 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 a carrier component. can

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, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane /may contain propellants such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, specifically water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid ester of sorbitan, and the like may be used.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, microcrystals Sex cellulose, aluminum metahydroxide, bentonite, agar, and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide as carrier components Ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative or ethoxylated glycerol fatty acid ester and the like can be used.

The cosmetic composition of the present invention can be prepared according to a method for preparing a cosmetic composition conventionally performed in the art, except that it contains an active ingredient that exhibits skin wrinkle improvement activity and skin whitening activity.

As described above, according to the present invention, it is possible to provide a composition for skin wrinkle improvement and a skin whitening composition using the extract of sagebrush.

Such a composition of the present invention can be commercialized as food, cosmetics, pharmaceuticals, and the like.

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> Preparation of extract

After leaching for 24 hours by adding 70% ethanol by weight of 20 times the weight of the sagebrush (outpost) powder, it was filtered through Whatman paper filter paper No. 2, and the filtrate was concentrated under reduced pressure to prepare an extract of sagebrush.

And after suspending and dissolving the obtained ethanol extract in distilled water (water) of 10 times the weight, hexane (n-hexane), dichloromethane (CH2Cl2), ethyl acetate (EtOAc) and butanol (BuOH) were sequentially fractionated to obtain a fraction prepared.

<Experimental Example> Elastase inhibitory activity test, tyrosinase inhibitory activity test, etc.

<Experimental Example 1> Elastase inhibitory activity test

After adding samples diluted by concentration to 50 mM Tris-Cl buffer (pH 8.5), 1 mg/ml of the substrate N-succinyl-(Ala) _3- p-nitroanilide was added to 50 mM Tris-Cl buffer (pH 8.5). 0.6U/ml PPE (porcine pancreas elastase) was added, mixed well, and reacted at 25°C, and absorbance was measured at 405 nm using an ELISA reader. By measuring the elastase inhibitory ability compared to the control group, which is the sample untreated group, IC ₅₀ , which is the 50% inhibitory concentration of elastase, was obtained. As a positive control, ursolic acid was used. The results are shown in Table 1 below. The results are the results of three repeated experiments for each concentration.

Elastase inhibitory activity (IC ₅₀ , μg/ml) division Elastase inhibition assay (μg/ml)) 70% Ethanol Extract <100 hexane fraction 88.16 dichloromethane fraction 20.21 ethyl acetate fraction 88.80 butanol fraction 28.89 residual water layer >1000 ursolic acid 46.29

As will be identified in the table, except for the remaining water layer and the embodiment extracts and fractions it is all clearly showed the elastase inhibitory activity, in particular dichloromethane fraction and the butanol fraction has the IC ₅₀ are respectively 20.21 ㎍ / ml, 28.89 ㎍ /ml showed high inhibitory activity. _{The IC 50} of ursolic acid as a positive control was 46.29 μg/ml.

<Experimental Example 2> Tyrosinase inhibitory activity test

After adding the sample diluted by concentration to 0.1 M potassium phosphate buffer (pH 6.5) and 1250 unit/ml mushroom-derived tyrosinase, add the substrate 1.5 mM L-tyrosine, mix well, and react at 37°C for 20 minutes, followed by 490 nm Absorbance was measured using an ELISA reader. By measuring the tyrosinase inhibitory ability compared to the control group, which is a sample untreated group, IC ₅₀ , which is a 50% inhibitory concentration of tyrosinase, was obtained. Arbutin was used as a positive control. The results are shown in Table 2 below. The results are the results of three repeated experiments for each concentration.

Tyrosinase inhibitory activity (IC ₅₀ , μg/ml) division Tyrosinase inhibition assay (㎍/ml)) 70% Ethanol Extract >1000 hexane fraction >1000 dichloromethane fraction 546.67 ethyl acetate fraction 255.65 butanol fraction <100 residual water layer >1000 arbutin 135.03

The table shows that the butanol fraction showed the highest tyrosinase inhibitory activity.

<Experimental Example 3> Anti-inflammatory activity test - NO production inhibition activity test in macrophages

To evaluate the intracellular anti-inflammatory activity of the extract of Example in RAW264.7 cells, which are mouse macrophages, the cells were ^{aliquoted at a cell number of 5×10 4} cells/well in a 96 well plate, and at 37° C., 5% CO ₂ for 24 hours. incubated during Samples were treated in a 96-well plate for 1 hour at each concentration, treated with LPS (Lipopolysacharide) 1㎍/ml, and cultured for 24 hours to evaluate anti-inflammatory activity. Anti-inflammatory activity evaluation through NO assay was performed using griess A and B reagents, and 1% sulfanilamide and 0.1% N-(1-naphtyl)ethylenediamine dihydrochloride reagent were mixed in a 1:1 ratio. After processing the sample in a 96 well plate, _{the supernatant of the medium cultured for 24 hours at 37°C and 5% CO 2} was mixed with griess (A+B) reagent in a 1:1 ratio, 50 μl each, and stored at room temperature for 10 minutes. _{Then, the IC 50} was obtained by measuring the NO production inhibitory ability compared to the control group, which is the untreated group, through ELISA measurement at 540 nm. The results are shown in Table 3 below. The results are the results of three repeated experiments for each concentration.

NO production inhibitory activity (IC ₅₀ , μg/ml) division NO production inhibitory activity (IC ₅₀ , μg/ml)) 70% Ethanol Extract 86.21 hexane fraction >100 dichloromethane fraction >100 ethyl acetate fraction 37.29 butanol fraction >100 residual water layer >100

The above table shows that the ethyl acetate fraction has the highest NO production inhibitory activity.

Meanwhile, cytotoxicity was measured by MTT method. Specifically, the remaining medium in the 96-well plate was removed, and 100 μl of serum free medium containing 10% of 5 mg/ml MTT solution was put into each well, and placed in an incubator at _{37° C. and 5% CO 2 to react for 2 hours.} After removing the medium, DMSO was added at 100 μl/well, dissolved in a shaker for 15 minutes, and absorbance was measured at 540 nm using an ELSA reader to determine the cell viability, thereby confirming the toxicity of the sample. It did not show any particular cytotoxicity at the highest treatment concentration of 100 μg/ml.

<Experimental Example 4> Antioxidant activity - DPPH method and ABST method experiment

<Experimental Example 4-1> DPPH method

After making a 200 uM DPPH methanol solution, add 100 μl of the DPPH solution and 100 μl of a sample diluted with methanol for each concentration, and react at room temperature for 30 minutes, and then measure the absorbance at 517 nm using an ELISA reader. DPPH scavenging activity was obtained compared with the untreated group. The results are shown in Table 4 below as _{a concentration IC 50} corresponding to a DPPH scavenging activity of 50%. As a positive control, BHA, an antioxidant, was used. The results are from three repeated experiments for each concentration.

DPPH radical scavenging rate (%) = (Control-sample) / Control × 100

Antioxidant activity according to DPPH method (IC ₅₀ , μg/ml) division DPPH assay (μg/ml)) 70% Ethanol Extract 24.3 hexane fraction 97.43 dichloromethane fraction 3.12 ethyl acetate fraction 3.77 butanol fraction 1.86 residual water layer >100 BHA 15.58

The results in the table above show that the butanol fraction has the highest DPPH scavenging activity.

<Experimental Example 2-2> ABTS method

7 mM ABTS buffer and 2.45 mM potassium persulfate buffer (K ₂ S ₂ O ₈ ) were prepared _{, mixed in ABTS:K 2} S ₂ O ₈ =2:1 ratio, and reacted under light blocking for 12-16 hours to form cationic radicals. After stock buffer was diluted so that the OD value measured at 734nm was 0.70±0.02, 100 μl of buffer and 100 μl of the extract sample diluted by concentration were added and reacted at room temperature for 6 minutes. After the reaction, absorbance was measured at 734 nm, and the ABTS cation radical scavenging activity was obtained compared to the control group, untreated group, according to the following equation. The results are shown in Table 5 below as _{the concentration IC 50} corresponding to 50% of DPPH scavenging activity. As a positive control, BHA, an antioxidant, was used. The results are from three repeated experiments for each concentration.

ABTS cation radical scavenging rate (%) = (Control-sample) / Control X 100

Antioxidant activity according to ABTS method (IC ₅₀ , μg/ml) division ABTS assay (μg/ml)) 70% Ethanol Extract 9.5 hexane fraction 94.20 dichloromethane fraction 10.83 ethyl acetate fraction 7.31 butanol fraction 14.83 residual water layer >100 BHA 15.58

The above table shows that the ethyl acetate fraction has the best ABTS cation radical scavenging activity.

Claims (12)

Including the extract of sagebrush as an active ingredient,
The extract is a cosmetic composition for improving skin wrinkles, characterized in that any one of the following (a) to (d):
(a) a dichloromethane fraction extract obtained by suspending a mixed solvent extract of water and ethanol in water and fractionating with dichloromethane;
(b) a butanol fraction extract obtained by suspending a mixed solvent extract of water and ethanol in water, followed by fractionation with butanol;
(c) a dichloromethane fractional extract obtained by suspending the mixed solvent extract of water and ethanol in water and sequentially fractionating it with hexane and dichloromethane as a fractionation solvent; and
(d) butanol fractional extract obtained by suspending the water and ethanol mixed solvent extract in water and sequentially fractionating it with hexane, dichloromethane, ethyl acetate and butanol as a fractionation solvent.
Including the extract of sagebrush as an active ingredient,
The extract is a food composition for improving skin wrinkles, characterized in that any one of the following (a) to (d):
(a) a dichloromethane fraction extract obtained by suspending a mixed solvent extract of water and ethanol in water and fractionating with dichloromethane;
(b) a butanol fraction extract obtained by suspending a mixed solvent extract of water and ethanol in water, followed by fractionation with butanol;
(c) a dichloromethane fractional extract obtained by suspending the mixed solvent extract of water and ethanol in water and sequentially fractionating it with hexane and dichloromethane as a fractionation solvent; and
(d) butanol fractional extract obtained by suspending the water and ethanol mixed solvent extract in water and sequentially fractionating it with hexane, dichloromethane, ethyl acetate and butanol as a fractionation solvent.
Including the extract of sagebrush as an active ingredient,
The extract is a cosmetic composition for skin whitening, characterized in that any one of the following (a) to (d):
(a) an ethyl acetate fractional extract obtained by suspending a mixed solvent extract of water and ethanol in water, followed by fractionation with dichloromethane;
(b) a butanol fraction extract obtained by suspending a mixed solvent extract of water and ethanol in water, followed by fractionation with butanol;
(c) an ethyl acetate fractional extract obtained by suspending the water and ethanol mixed solvent extract in water and sequentially fractionating it with hexane, dichloromethane, and ethyl acetate as a fractionation solvent; and
(d) butanol fractional extract obtained by suspending the water and ethanol mixed solvent extract in water and sequentially fractionating it with hexane, dichloromethane, ethyl acetate and butanol as a fractionation solvent.
Including the extract of sagebrush as an active ingredient,
The extract is a food composition for skin whitening, characterized in that any one of the following (a) to (d):
(a) an ethyl acetate fractional extract obtained by suspending a mixed solvent extract of water and ethanol in water, followed by fractionation with dichloromethane;
(b) a butanol fraction extract obtained by suspending a mixed solvent extract of water and ethanol in water, followed by fractionation with butanol;
(c) an ethyl acetate fractional extract obtained by suspending the water and ethanol mixed solvent extract in water and sequentially fractionating it with hexane, dichloromethane, and ethyl acetate as a fractionation solvent; and
(d) butanol fractional extract obtained by suspending the water and ethanol mixed solvent extract in water and sequentially fractionating it with hexane, dichloromethane, ethyl acetate and butanol as a fractionation solvent.
Including the extract of sagebrush as an active ingredient,
The extract is an anti-inflammatory pharmaceutical composition, characterized in that any one of the following (a) and (b):
(a) a fractional extract obtained by suspending a mixed solvent extract of water and ethanol in water, followed by fractionation with ethyl acetate; and
(b) an ethyl acetate fractional extract obtained when a mixed solvent extract of water and ethanol is suspended in water and then sequentially fractionated with hexane, dichloromethane and ethyl acetate.
6. The method of claim 5,
The composition, characterized in that the mixed solvent of water and ethanol is 70% ethanol.
Including the extract of sagebrush as an active ingredient,
The extract is an anti-inflammatory food composition, characterized in that any one of (a) and (b):
(a) a fractional extract obtained by suspending a mixed solvent extract of water and ethanol in water, followed by fractionation with ethyl acetate; and
(b) an ethyl acetate fractional extract obtained when a mixed solvent extract of water and ethanol is suspended in water and then sequentially fractionated with hexane, dichloromethane and ethyl acetate.
8. The method of claim 7,
The composition, characterized in that the mixed solvent of water and ethanol is 70% ethanol.
Including the extract of sagebrush as an active ingredient,
The extract is a cosmetic composition for inhibiting inflammatory skin irritation, characterized in that any one of the following (a) and (b):
(a) a fractional extract obtained by suspending a mixed solvent extract of water and ethanol in water, followed by fractionation with ethyl acetate; and
(b) an ethyl acetate fractional extract obtained when a mixed solvent extract of water and ethanol is suspended in water and then sequentially fractionated with hexane, dichloromethane and ethyl acetate.
10. The method of claim 9,
The composition, characterized in that the mixed solvent of water and ethanol is 70% ethanol.
Including the extract of sagebrush as an active ingredient,
The extract is a pharmaceutical composition for improving skin hyperpigmentation, characterized in that any one of the following (a) to (d):
(a) an ethyl acetate fractional extract obtained by suspending a mixed solvent extract of water and ethanol in water, followed by fractionation with dichloromethane;
(b) a butanol fraction extract obtained by suspending a mixed solvent extract of water and ethanol in water, followed by fractionation with butanol;
(c) an ethyl acetate fractional extract obtained by suspending the water and ethanol mixed solvent extract in water and sequentially fractionating it with hexane, dichloromethane, and ethyl acetate as a fractionation solvent; and
(d) a butanol fraction extract obtained by suspending the water and ethanol mixed solvent extract in water and sequentially fractionating it with hexane, dichloromethane, ethyl acetate and butanol as a fractionation solvent.

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