KR20220083148A - Cosmetic composition comprising Potentilla supina extract as an active ingredient - Google Patents

Abstract

The present invention relates to a composition for reducing skin moisture content and preventing or improving pigmentation, etc., comprising the gaeso shirangae extract as an active ingredient. It is expected to be widely used as an improvement, prevention, or treatment for skin problems.

Description

Cosmetic composition comprising Potentilla supina extract as an active ingredient}

The present invention relates to a composition for preventing or improving at least one selected from the group consisting of a skin moisture content decrease and pigmentation, and the like, comprising an extract of Gaeso Shirangaebi extract as an active ingredient.

The skin protects internal organs from external environmental stimuli and plays an important role in maintaining body homeostasis, such as thermoregulation. As the skin ages, it is accompanied by external changes such as a decrease in skin elasticity, a change in skin color, and the formation of skin wrinkles. This skin aging process appears as a result of the complex action of intrinsic aging (or natural aging) that occurs naturally as we age, and photoaging in the surrounding environment, which is extrinsic aging, particularly ultraviolet (UV) light. Although there are differences in the mechanism of wrinkle generation between the two types of aging, in the skin, the proliferation or metabolism of epidermal cells is reduced, the bond between the epidermis and the dermis is weak, so it is easy to be damaged by even a small stimulus, and the decrease in collagen synthesis and excessive Physiological changes such as increased expression of MMPs (Matrix Metalloproteinases), which are collagen degrading enzymes, appear, and the skin becomes dry, and the skin loses elasticity and appears sagging due to structural changes in the epidermis and dermis, and wrinkles gradually deepen. Although natural aging is unavoidable in skin aging, effectively blocking photoaging caused by ultraviolet rays, an environmental factor, is a way to delay and suppress skin aging.

Recently, ozone layer depletion due to environmental pollution has increased the amount of UV rays, and accordingly, research on photoaging is attracting attention. Appearance features such as roughness, loss of elasticity, and wrinkles are observed in photoaging skin, and the main research area of photoaging is on changes in skin wrinkles. A number of research results have been reported on basic physiological metabolic changes such as synthesis, decomposition, and moisture content of collagen, which is a major component of the skin, in relation to skin wrinkle formation due to photoaging.

A person's skin color is determined by the concentration and distribution of melanin inside the skin. Melanin is synthesized in melanocytes in the epidermal layer of the skin. In the melanosome, an organelle in melanocytes, an enzyme called tyrosinase acts on tyrosine, a type of amino acid, to produce DOPA and dopaquinone. ) and then undergoes a non-enzymatic oxidation reaction. When such melanin synthesis occurs in the skin excessively, the skin tone is darkened, and spots, freckles, and the like occur. Therefore, by inhibiting the tyrosinase activity in the skin to inhibit the synthesis of melanin, not only can the skin tone be brightened to realize skin whitening, but also hyperpigmentation of the skin such as spots and freckles caused by UV rays, hormones and genetic causes It can improve sedation.

On the other hand, recently, as a number of cases in which side effects occur due to chemical components of cosmetics are known, consumers' interest in cosmetics manufactured by containing natural ingredients is increasing. Accordingly, many cosmetics using natural products have been developed to reduce skin irritation caused by various chemicals. Natural ingredients have fewer side effects on the skin, and as consumers' response to cosmetics using natural ingredients increases, the value of development as a cosmetic raw material is further increasing. However, little is known about whether or not natural raw sage ragweed is effective in reducing skin moisture content and improving pigmentation.

Korean Patent Registration No. 10-1961152

Accordingly, the present inventors have completed the present invention by confirming that the gaeso shirangaebi extract has the effect of lowering skin moisture content and improving pigmentation while not having cytotoxicity.

Accordingly, it is an object of the present invention to provide a composition for preventing or improving at least one selected from the group consisting of reduced skin moisture content and pigmentation, comprising the extract of Gaeso Shirangaebi extract as an active ingredient.

Another object of the present invention is to provide a composition for skin whitening comprising an extract of Gaeso Shirangaebi as an active ingredient.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.

In order to achieve the object of the present invention, the present invention provides a composition for preventing or improving at least one selected from the group consisting of reduced skin moisture content and pigmentation, comprising the extract of Gaeso Shirangaebi as an active ingredient.

In addition, the present invention provides a composition for skin whitening comprising the extract of gaeso shirangaebi as an active ingredient.

In one embodiment of the present invention, the composition may be a pharmaceutical composition, a cosmetic composition, or a food composition, but is not limited thereto.

In another embodiment of the present invention, the extract is water, an alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, It may be an extract using one or more solvents selected from the group consisting of methylene chloride, hexane, cyclohexane, petroleum ether, subcritical fluid, and supercritical fluid, but is limited thereto it is not going to be

In another embodiment of the present invention, the gaeso shirangae extract may reduce MITF (microphthalmia-associated transcription factor) expression or activity, but is not limited thereto.

In another embodiment of the present invention, the Gaesosirangae extract may reduce the expression or activity of tyrosinase, but is not limited thereto.

In another embodiment of the present invention, the Gaesosirangae extract may reduce the expression or activity of TYRP-1 (Tyrosinase-related protein 1) or TYRP-2 (Tyrosinase-related protein 2), but is limited thereto not.

In another embodiment of the present invention, the Gaesosirangae extract can increase the expression or activity of one or more moisturizing factors selected from the group consisting of hyaluronan synthase-1 (HAS-1), HAS-2, and HAS-3. However, the present invention is not limited thereto.

In another embodiment of the present invention, the Gaesosirangae extract can reduce the expression or activity of one or more selected from the group consisting of HYAL-1 (Hyaluronidase-1), HYAL-2, HYAL-3, and HYAL-4. However, the present invention is not limited thereto.

In another embodiment of the present invention, the Gaesosirangae extract may inhibit melanin secretion or synthesis, but is not limited thereto.

In another embodiment of the present invention, the composition for whitening may prevent, improve, or treat pigmentation disorders, but is not limited thereto.

In another embodiment of the present invention, the skin pigmentation disease is at least one selected from the group consisting of melasma, freckles, lentigines, nevus, drug-induced pigmentation, post-inflammatory pigmentation, and hyperpigmentation occurring in dermatitis. However, the present invention is not limited thereto.

In addition, the present invention provides a method for preventing or improving at least one selected from the group consisting of reduced skin moisture content and pigmentation, comprising administering to an individual a composition comprising an extract of Gaeso Shirangaebi extract as an active ingredient.

In addition, the present invention provides a use for preventing or improving at least one selected from the group consisting of reduced skin moisture content and pigmentation of a composition comprising an extract of Gaeso Shirangaebi extract as an active ingredient.

In addition, the present invention provides a use for producing one or more preventive or ameliorating medicaments, cosmetics, or foods selected from the group consisting of skin moisture content reduction and pigmentation of the gaeso shirangae extract.

In addition, the present invention provides a method for treating a skin pigmentation disease comprising administering to an individual a composition comprising an extract of gaeso shirangaebi as an active ingredient.

In addition, the present invention provides a use for the prevention and / or treatment of skin pigmentation diseases of a composition comprising the extract of Gaeso Shirangaebi as an active ingredient.

In addition, the present invention provides a use for producing a medicament used in skin pigmentation diseases of the extract of Gaeso Shirangaebi extract.

The present invention relates to a composition for preventing or improving skin moisture content and for preventing or improving pigmentation, and the like, comprising an extract of Gaeso Shirangae as an active ingredient. It is expected to be widely used as an improvement, prevention, or treatment for skin problems.

1 shows the results of confirming the ROS reduction effect on B16F10 cells of an ethanol extract of Potentilla supina (hereinafter Ps-EE).
Figure 2a shows the result of confirming the cytotoxicity of Ps-EE to B16F10 cells.
Figure 2b shows the results of confirming the cytotoxicity of Ps-EE to HaCaT cells.
Figure 3a shows the effect of reducing melanin secretion by α-MSH treatment of Ps-EE.
Figure 3b shows the effect of reducing melanin formation by α-MSH treatment of Ps-EE.
Figure 4 shows the effect of Ps-EE on the mRNA expression of Tyrosinase, TYRP-1, TYRP-2, and MITF, which are known to be related to melanogenesis in the context of α-MSH treatment.
Figure 5 shows the effect of Ps-EE on the mRNA expression of moisture-related genes HAS-1, HAS-2, and HAS-3.
6 shows the effect of Ps-EE on the mRNA expression of genes HYAL-1, HYAL-2, HYAL-3, and HYAL-4 related to the degradation of moisturizing factors.

The present invention provides a composition for preventing or improving at least one selected from the group consisting of reduced skin moisture content and pigmentation, comprising the extract of Gaeso Shirangaebi as an active ingredient.

In addition, the present invention provides a composition for whitening comprising the extract of Gaeso shirangaebi as an active ingredient.

Hereinafter, the present invention will be described in detail.

Potentilla supina of the present invention may be preferably, but is not limited thereto.

Gaesosirangaebi, also called Gipsosrangaebi, is a perennial herb belonging to the Primrose family. It is native to northern Africa and Eurasia. In Korea, it grows in fields, roadsides, and clearings all over the country. The stem height is about 20-50 cm. The bottom part grows obliquely to the side, and then stands upright. The leaves are opposite, pinnate compound leaves composed of 2-4 pairs of small leaves, and the petiole is long. Small leaves are obovate, oval or lanceolate, 0.7-2.5 cm long and 0.8-1.4 cm wide, with a rounded tip and serrated edges. From May to September, several yellow flowers with a diameter of 6-8 mm hang in the leaf axil at the tip of the branch. The fruit is a fruit.

Gaeso Shirangaebi of the present invention can be used as it is without damaging its original form, or after performing a pretreatment process in consideration of the process speed and process (manufacturing) efficiency intended by those skilled in the art, the pretreatment process includes, for example, Conventional screening, washing, cutting, powdering, drying, etc. steps may be performed.

The extract of Gaeso Shirangaegi of the present invention can be obtained from the whole plant or a part of a plant, for example, it can be obtained from a site comprising one or more selected from the group consisting of stems, roots, leaves, fruits, and flowers.

Gaeso shirangaebi extract of the present invention can be extracted by a known natural extracting method. For example, extraction may be performed with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, and a subcritical or supercritical fluid. The organic solvent having 1 to 6 carbon atoms is alcohol, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, having 1 to 6 carbon atoms. chloride), hexane, cyclohexane, and petroleum ether may be at least one selected from the group consisting of, but is not limited thereto.

The extract of Gaeso Shirangaebi of the present invention may be preferably extracted with ethanol, but is not limited thereto. As the extraction method, conventional extraction methods such as cold extraction, warm soaking, and heating using the above solvent can be used.

The ethanol concentration is not limited, for example, 30 to 100%, 40 to 100%, 50 to 100%, 60 to 100%, 60 to 90%, 60 to 80%, 70 to 90%, 70 to It may be 100%, 80-100%, 90-100% ethanol extract.

In the present invention, the gaeso shirangae extract may reduce MITF (microphthalmia-associated transcription factor) expression or activity, but is not limited thereto.

In the present invention, the Gaesosirangae extract may reduce the expression or activity of tyrosinase, but is not limited thereto.

In the present invention, the gaeso sirangae extract may reduce the expression or activity of TYRP-1 (Tyrosinase-related protein 1) or TYRP-2 (Tyrosinase-related protein 2), but is not limited thereto.

In the present invention, the Gaesosirangae extract can increase the expression or activity of one or more moisturizing factors selected from the group consisting of HAS-1 (hyaluronan synthase-1), HAS-2, and HAS-3, but is limited thereto. it is not

In the present invention, the Gaeso Shirangae extract can reduce the expression or activity of one or more selected from the group consisting of HYAL-1 (Hyaluronidase-1), HYAL-2, HYAL-3, and HYAL-4, but is limited thereto it is not

In the present invention, the Gaesosirangae extract may inhibit melanin secretion or synthesis, but is not limited thereto.

In one embodiment of the present invention, as a result of verifying the activity of the ethanol extract of Gaeso sirangaebi, the extract has no cytotoxicity (see Example 4), has an excellent antioxidant effect (see Example 3), and inhibits melanin formation and secretion Reduces (see Example 5), whitening-related factors MITF, tyrosinase, TYRP-1 and TYRP-2, and moisturizing-related genes HAS-1 (hyaluronan synthase-1), HAS-2 and HAS-3 It was confirmed that HYAL-1, HYAL-2, HYAL-3, and HYAL-4 related to decomposition of moisturizing factors decreased (see Examples 6 and 7).

Accordingly, the gaeso shirangae extract of the present invention can be used as an active ingredient in a cosmetic, medicament, quasi-drug, food or health functional food composition for preventing and improving at least one selected from the group consisting of reduced skin moisture content and pigmentation. .

In the present invention, 'skin moisturizing' or 'improving skin moisture content reduction' refers to promoting the differentiation of keratinocytes to inhibit or inhibit the decrease in moisture in the skin or increase the moisture content of the skin to smooth the skin surface, It is said to give shine.

In the present invention, 'whitening' refers to not only brightening skin tone by inhibiting the synthesis of melanin, but also improving skin hyperpigmentation (spots and freckles, etc.) caused by various causes such as UV rays, hormones, or heredity. say that

In the present invention, 'pigmentation' may mean a pigmentation disease caused by overproduction or/and (excessive) accumulation of melanin. The (hyper-pigmentation disease) includes all lesions and symptoms in which the condition is progressing in relation to the melanin mechanism in addition to the skin. Although not limited thereto, in one embodiment, the pigmentation disease may be a skin pigmentation disease.

The composition according to the present invention is a pathological condition of excessive melanin pigmentation, for example, aging/photoaging, rapid hormonal changes such as pregnancy, skin damage and regeneration due to wounds, inflammations, burns, etc. pigmentation, blemishes, It can be used to improve and alleviate freckles, blemishes, spots, spots, nevus ota, age spots, senile lupus, melanin dermatosis, etc.

In the present invention, the skin pigmentation disease may be at least one selected from the group consisting of melasma, freckles, black spots, nevus, drug-induced pigmentation, post-inflammatory pigmentation, and hyperpigmentation occurring in dermatitis, but limited thereto it is not going to be

The nevus is specifically nevus flat, nevus pigmentosa, Becker's nevus, Nevus Spilus, nevus of Ota, Acquired bilateral nevus of nevus of Ota. Ota-like macules), Nevus of Ito, Blue nevus, Melanocytic nevus, Junctional nevus, Compound nevus, Intradermal nevus ), Halo nevus, Congenital nevocytic nevus, Spitz nevus, and Dysplastic nevus.

Melanin is a pigment component widely distributed in living organisms such as skin, hair, and pupils, and is synthesized from melanosomes in melanocytes of the human epidermal layer. Tyrosine is initiated by the enzyme tyrosinase. As a substance, melanin is biosynthesized by oxidation and polymerization to be converted into DOPA (3,4-dihydroxy-phenylalanine) or DOPA quinone. Biosynthesized melanin increases resistance to skin stimuli such as UV rays, but excessive melanin synthesis causes pigmentation such as spots, freckles, and age spots, so measurement of tyrosinase activity is accepted as an important indicator of skin whitening effect. .

In an embodiment of the present invention, it was confirmed that the gaeso sirangae extract significantly reduced melanin secretion and production within a range that did not cause toxicity to cells, and could inhibit the expression and activity of melanin production-related factors.

The content of the extract of Gaeso Shirangae in the composition of the present invention can be appropriately adjusted according to the symptoms of the disease, the degree of progression of the symptoms, the condition of the patient, etc., for example, 0.0001 to 99.9% by weight, or 0.001 to 50, based on the total weight of the composition. It may be a weight %, but is not limited thereto. The content ratio is a value based on the dry amount from which the solvent is removed.

The pharmaceutical composition according to the present invention may further include suitable carriers, excipients and diluents commonly used in the preparation of pharmaceutical compositions. The excipient may be, for example, at least one selected from the group consisting of a diluent, a binder, a disintegrant, a lubricant, an adsorbent, a humectant, a film-coating material, and a controlled-release additive.

The pharmaceutical composition according to the present invention can be prepared according to a conventional method, respectively, in powders, granules, sustained-release granules, enteric granules, liquids, eye drops, elsilic, emulsions, suspensions, alcohols, troches, fragrances, and limonaade. , tablets, sustained release tablets, enteric tablets, sublingual tablets, hard capsules, soft capsules, sustained release capsules, enteric capsules, pills, tinctures, soft extracts, dry extracts, fluid extracts, injections, capsules, perfusates, Warnings, lotions, pasta, sprays, inhalants, patches, sterile injection solutions, or external preparations such as aerosols can be formulated and used, and the external preparations are creams, gels, patches, sprays, ointments, warning agents , lotion, liniment, pasta, or cataplasma.

Carriers, excipients and diluents that may be included in the pharmaceutical composition according to the present invention include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In the case of formulation, it is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

Corn starch, potato starch, wheat starch, lactose, sucrose, glucose, fructose, di-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, phosphoric acid as additives for tablets, powders, granules, capsules, pills, and troches according to the present invention Calcium monohydrogen, calcium sulfate, sodium chloride, sodium hydrogen carbonate, purified lanolin, microcrystalline cellulose, dextrin, sodium alginate, methyl cellulose, sodium carboxymethyl cellulose, kaolin, urea, colloidal silica gel, hydroxypropyl starch, hydroxypropyl methyl excipients such as cellulose, 1928, 2208, 2906, 2910, propylene glycol, casein, calcium lactate, and Primogel; Gelatin, gum arabic, ethanol, agar powder, cellulose acetate phthalate, carboxymethylcellulose, calcium carboxymethylcellulose, glucose, purified water, sodium caseinate, glycerin, stearic acid, sodium carboxymethylcellulose, sodium methylcellulose, methylcellulose, microcrystalline cellulose, dextrin , hydroxycellulose, hydroxypropyl starch, hydroxymethylcellulose, purified shellac, starch powder, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, etc. Hydroxypropyl methylcellulose, corn starch, agar powder, methylcellulose, bentonite, hydroxypropyl starch, sodium carboxymethylcellulose, sodium alginate, calcium carboxymethylcellulose, calcium citrate, sodium lauryl sulfate, silicic anhydride, 1-hydroxy Propyl cellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde treated casein and gelatin, alginic acid, amylose, guar gum, sodium bicarbonate, polyvinylpyrrolidone, calcium phosphate, gelled starch, gum arabic, Disintegrants such as amylopectin, pectin, sodium polyphosphate, ethyl cellulose, sucrose, magnesium aluminum silicate, di-sorbitol solution, light anhydrous silicic acid; Calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopodite, kaolin, petrolatum, sodium stearate, cacao butter, sodium salicylate, magnesium salicylate, polyethylene glycol 4000, 6000, liquid paraffin, hydrogenated soybean oil (Lubri) wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, macrogol, synthetic aluminum silicate, silicic anhydride, higher fatty acid, higher alcohol, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, starch, sodium chloride, A lubricant such as sodium acetate, sodium oleate, dl-leucine, light anhydrous silicic acid; may be used.

As additives for the liquid formulation according to the present invention, water, diluted hydrochloric acid, diluted sulfuric acid, sodium citrate, monostearate sucrose, polyoxyethylene sorbitol fatty acid esters (Twinester), polyoxyethylene monoalkyl ethers, lanolin ethers, Lanolin esters, acetic acid, hydrochloric acid, aqueous ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethyl cellulose, sodium carboxymethyl cellulose, etc. can be used.

In the syrup according to the present invention, a sucrose solution, other sugars or sweeteners may be used, and if necessary, a fragrance, colorant, preservative, stabilizer, suspending agent, emulsifying agent, thickening agent, etc. may be used.

Purified water may be used in the emulsion according to the present invention, and if necessary, an emulsifier, preservative, stabilizer, fragrance, etc. may be used.

Suspending agents such as acacia, tragacantha, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose (HPMC), HPMC1828, HPMC2906, HPMC2910 may be used in the suspending agent according to the present invention. and, if necessary, surfactants, preservatives, stabilizers, colorants, and fragrances may be used.

The injection according to the present invention includes distilled water for injection, 0.9% sodium chloride injection solution, ring gel injection solution, dextrose injection solution, dextrose + sodium chloride injection solution, PEG (PEG), lactated ring gel injection solution, ethanol, propylene glycol, non-volatile oil-sesame oil , solvents such as cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristate, and benzene benzoate; Solubilizing aids such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethyl acetamide, butazolidine, propylene glycol, tweens, nijeongtinamide, hexamine, and dimethyl acetamide; Weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), organic compounds, proteins, buffers such as albumin, peptone, gum; isotonic agents such as sodium chloride; Stabilizers such as sodium bisulfite (NaHSO ₃ ) carbon dioxide gas, sodium metabisulfite (Na ₂ S ₂ O ₃ ), sodium sulfite (Na ₂ SO ₃ ), nitrogen gas (N ₂ ), ethylenediaminetetraacetic acid; sulphating agents such as sodium bisulfide 0.1%, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetate, acetone sodium bisulfite; analgesic agents such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; suspending agents such as SiMC sodium, sodium alginate, Tween 80, or aluminum monostearate.

The suppository according to the present invention includes cacao fat, lanolin, witepsol, polyethylene glycol, glycerogelatin, methyl cellulose, carboxymethyl cellulose, a mixture of stearic acid and oleic acid, Subanal, cottonseed oil, peanut oil, palm oil, cacao butter + Cholesterol, Lecithin, Lanet Wax, Glycerol Monostearate, Tween or Span, Imhausen, Monolene (Propylene Glycol Monostearate), Glycerin, Adeps Solidus, Butyrum Tego -G), Cebes Pharma 16, Hexalide Base 95, Cotomar, Hydroxote SP, S-70-XXA, S-70-XX75 (S-70-XX95), Hydro Hydrokote 25, Hydrokote 711, Idropostal, Massa estrarium, A, AS, B, C, D, E, I, T, Massa-MF, Masupol, Masupol-15, Neosupostal-N, Paramound-B, Suposiro (OSI, OSIX, A, B, C, D, H, L), Suppository IV type (AB, B, A, BC, BBG, E, BGF, C, D, 299), supostal (N, Es), Wecobi (W, R, S, M, Fs), tester triglyceride base (TG-95, MA, 57) and The same mechanism may be used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the extract, for example, starch, calcium carbonate, sucrose ) or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

Liquid formulations for oral administration include suspensions, internal solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the type, severity, drug activity, and type of the patient's disease; Sensitivity to the drug, administration time, administration route and excretion rate, treatment period, factors including concurrent drugs and other factors well known in the medical field may be determined.

The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or may be administered in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. In consideration of all of the above factors, it is important to administer an amount capable of obtaining the maximum effect with a minimum amount without side effects, which can be easily determined by a person skilled in the art to which the present invention pertains.

The pharmaceutical composition of the present invention may be administered to an individual by various routes. All modes of administration can be contemplated, for example, oral administration, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, paraspinal space (intrathecal) injection, sublingual administration, buccal administration, rectal insertion, vaginal It can be administered according to internal insertion, ocular administration, ear administration, nasal administration, inhalation, spraying through the mouth or nose, skin administration, transdermal administration, and the like.

The pharmaceutical composition of the present invention is determined according to the type of drug as an active ingredient along with several related factors such as the disease to be treated, the route of administration, the patient's age, sex, weight, and the severity of the disease.

In the present invention, "individual" means a subject in need of treatment for a disease, and more specifically, human or non-human primates, mice, rats, dogs, cats, horses, cattle, etc. It may be a mammal of, but is not limited thereto.

In the present invention, "administration" means providing a given composition of the present invention to a subject by any suitable method.

In the present invention, "prevention" means any action that inhibits or delays the onset of a desired disease, and "treatment" means that the desired disease and metabolic abnormalities are improved or It means all actions that are advantageously changed, and "improvement" means all actions that reduce the parameters related to the desired disease, for example, the degree of symptoms by administration of the composition according to the present invention.

In addition, the present invention provides a food composition comprising the extract of Gaeso shirangaebi as an active ingredient.

In the present invention, food includes functional food and health functional food.

Preferably, the health functional food has the advantage of having a more excellent effect by ingesting it in the form of inner beauty food. The inner beauty is a food called 'eating cosmetics or beauty food'. You can choose and consume inner beauty foods that are suitable for each individual by considering your skin condition and lifestyle. More preferably, when a cosmetic containing the cosmetic composition and an inner beauty food containing the extract of the present invention are mixed, at least one selected from the group consisting of reduced skin moisture content and pigmentation compared to using only cosmetics or pharmaceuticals It has the advantage of being able to see a more effective skin improvement effect as the prevention or improvement effect is significantly increased.

When the extract of the present invention is used as a food additive, it can be added as it is or used together with other foods or food ingredients, and can be appropriately used according to a conventional method. The mixed amount of the active ingredient may be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment). In general, in the production of food or beverage, the extract of the present invention may be added in an amount of 15% by weight or less, or 10% by weight or less based on the raw material. However, for health and hygiene purposes or health control In the case of long-term ingestion for the purpose of

There is no particular limitation on the type of the food. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, There are alcoholic beverages and vitamin complexes, and includes all health functional foods in the ordinary sense.

The health beverage composition according to the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients, as in a conventional beverage. The above-mentioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetener, natural sweeteners such as taumartin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like can be used. The proportion of the natural carbohydrate is generally about 0.01-0.20 g, or about 0.04-0.10 g per 100 mL of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, Carbonating agents used in carbonated beverages, etc. may be contained. In addition, the composition of the present invention may contain the pulp for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These components may be used independently or in combination. The proportion of these additives is not critical, but is generally selected in the range of 0.01-0.20 parts by weight per 100 parts by weight of the composition of the present invention.

In addition, the present invention provides a cosmetic composition comprising an extract of gaeso shirangaebi as an active ingredient.

The formulation of the cosmetic composition according to the present invention includes skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, foundation, essence, nourishing essence, It may be in the form of a pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion or body cleanser.

The cosmetic composition of the present invention may further include a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, high molecular weight polysaccharides, and sphingolipids.

As the water-soluble vitamin, any one can be used as long as it can be formulated in cosmetics. Preferably, vitamin B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, nicotinic acid amide, folic acid, vitamin C, vitamin H, etc. are mentioned. Also, their salts (thiamine hydrochloride, sodium ascorbate salt, etc.) and derivatives (ascorbic acid-2-phosphate sodium salt, ascorbic acid-2-phosphate magnesium salt, etc.) are also included in the water-soluble vitamins that can be used in the present invention. Included. Water-soluble vitamins can be obtained by conventional methods, such as a microbial transformation method, a purification method from a microbial culture, an enzymatic method, or a chemical synthesis method.

The oil-soluble vitamin may be any as long as it can be formulated in cosmetics, but preferably includes vitamin A, carotene, vitamin D2, vitamin D3, vitamin E (d1-alpha tocopherol, d-alpha tocopherol, d-alpha tocopherol), etc. , their derivatives (ascorbine palmitate, ascorbine stearate, ascorbine dipalmitate, dl-alpha tocopherol acetate, dl-alpha tocopherol nicotinic acid, vitamin E, DL-pantothenyl alcohol, D-pantothenyl alcohol, pantothenyl ethyl ethers, etc.) are also included in the oil-soluble vitamins used in the present invention. Oil-soluble vitamins can be obtained by conventional methods such as a microbial transformation method, a purification method from a microbial culture, and an enzyme or chemical synthesis method.

The macromolecular peptide may be any type of peptide as long as it can be formulated in cosmetics. Preferably, collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, keratin, and the like are mentioned. Polymeric peptides can be purified and obtained by conventional methods such as purification from a microbial culture solution, enzyme method or chemical synthesis method, or can be purified and used from natural products such as dermis of pigs or cattle, silk fiber of silkworms, etc.

The high molecular weight polysaccharide may be any compound as long as it can be formulated in cosmetics, but preferably hydroxyethyl cellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate, or a salt thereof (such as sodium salt). For example, chondroitin sulfate or a salt thereof can be used after purification from mammals or fish.

The sphingolipid may be any as long as it can be formulated in cosmetics, and ceramide, phytosphingosine, sphingoglycolipid and the like are preferred. The sphingolipids can be purified by conventional methods from mammals, fish, shellfish, yeast or plants, or can be obtained by chemical synthesis.

In the cosmetic composition of the present invention, in addition to the above essential ingredients, other ingredients usually blended in cosmetics may be blended as needed.

Other ingredients that may be added include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, bactericides, antioxidants, plant extracts, pH adjusters, alcohols, colorants, fragrances, A blood circulation promoter, a cooling agent, a restrictive agent, purified water, etc. are mentioned.

Examples of the fats and oils include ester fats and oils, hydrocarbon oils, silicone oils, fluorine oils, animal fats, and vegetable oils.

As ester-based fats and oils, tri2-ethylhexanoate glyceryl, 2-ethylhexanoate cetyl, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, stearic acid Butyl, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldodecyl myristate, isocetyl isostearate, diethyl sebacate, adipine Diisopropyl acid, isoalkyl neopentanoate, tri(caprylic acid, capric acid) glyceryl, trimethylol propane tri-2-ethylhexanoate, trimethylol propane triisostearate, pentaelysitol tetra2-ethylhexanoate , cetyl caprylate, decyl laurate, hexyl laurate, decyl myristate, myristyl myristate, cetyl myristate, stearyl stearate, decyl oleate, cetyl ricinoleate, isostea laurate Lil, isotridecyl myristate, isocetyl palmitate, octyl stearate, isocetyl stearate, isodecyl oleate, octyldodecyl oleate, octyldodecyl linoleate, isostearic acid isopropyl, 2-ethylhexanoate cetoster Aryl, 2-ethyl stearyl hexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicapric acid, di(caprylic acid, capric acid) propylene glycol, dicaprylic acid propylene glycol, dicapric acid Neopentyl glycol phosphate, neopentyl glycol dioctanoate, glyceryl tricaprylate, glyceryl triundecyl, glyceryl triisopalmitate, glyceryl triisostearate, octyldodecyl neopentanoate, isostearyl octanoate , isononanoate octyl, neodecanoate hexyldecyl, neodecanoate octyldodecyl, isostearate isocetyl, isostearyl isostearate, isostearic acid octyldecyl, polyglycerol oleate, polyglycerol isostearate, Triisocetyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate, triethyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, dimalate Isostearyl, 2-ethylhexyl hydroxystearate, di2-ethylhexyl succinate, diisobutyl adipate, diisopropyl sebacate, dioctyl sebacate, Cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, phytsteryl isostearate, phytsteryl oleate, 12-stealoyl Ester type, such as hydroxy stearate isocetyl, 12-stealoyl hydroxy stearyl stearate, and 12-stealoyl hydroxy stearate isostearyl, etc. are mentioned.

Examples of hydrocarbon-based fats and oils include hydrocarbon-based fats and oils such as squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybudene, microcrystalline wax, petrolatum.

Examples of silicone-based oils and fats include polymethyl silicone, methylphenyl silicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane/methylcetyloxysiloxane copolymer, dimethylsiloxane/methylstealloxysiloxane copolymer, and alkyl Modified silicone oil, amino modified silicone oil, etc. are mentioned.

Perfluoropolyether etc. are mentioned as fluorine-type fats and oils.

As animal or vegetable oils, avocado oil, almond oil, olive oil, sesame oil, rice bran oil, saffron oil, soybean oil, corn oil, rapeseed oil, passerine oil, palm kernel oil, palm oil, castor oil, sunflower oil, grape seed oil , Cottonseed Oil, Palm Oil, Kukui Nut Oil, Wheat Germ Oil, Rice Germ Oil, Shea Butter, Wormwood Colostrum, Marcus Damien Nut Oil, Meadow Oil, Egg Yolk Oil, Tallow Oil, Horse Oil, Mink Oil, Orange Raffy Oil, Jojoba Oil , animal or plant oils and fats such as candelabra wax, carnauba wax, liquid lanolin, and hydrogenated castor oil.

Examples of the moisturizing agent include a water-soluble low-molecular moisturizer, a fat-soluble molecular moisturizer, a water-soluble polymer, and a fat-soluble polymer.

As water-soluble low molecular weight moisturizers, serine, glutamine, sorbitol, mannitol, pyrrolidone-sodium carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B (polymerization degree n = 2 or more), polypropylene Glycol (polymerization degree n=2 or more), polyglycerol B (polymerization degree n=2 or more), lactic acid, lactic acid salt, etc. are mentioned.

Cholesterol, cholesterol ester, etc. are mentioned as a fat-soluble low molecular weight humectant.

Examples of the water-soluble polymer include carboxyvinyl polymer, polyaspartate, tragacanth, xanthan gum, methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, water-soluble chitin, chitosan, and dextrin. can

Examples of the fat-soluble polymer include polyvinylpyrrolidone/eicocene copolymer, polyvinylpyrrolidone/hexadecene copolymer, nitrocellulose, dextrin fatty acid ester, and high molecular weight silicone.

Examples of the emollient include long-chain acylglutamic acid cholesteryl ester, hydroxystearic acid cholesteryl, 12-hydroxystearic acid, stearic acid, rosin acid, and lanolin fatty acid cholesteryl ester.

Nonionic surfactants, anionic surfactants, cationic surfactants, amphoteric surfactants, etc. are mentioned as surfactant.

Nonionic surfactants include self-emulsifying glycerin monostearate, propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerol fatty acid ester, sorbitan fatty acid ester, POE (polyoxyethylene) sorbitan fatty acid ester, POE sorbitan fatty acid ester, POE Glycerin fatty acid ester, POE alkyl ether, POE fatty acid ester, POE hydrogenated castor oil, POE castor oil, POE/POP (polyoxyethylene/polyoxypropylene) copolymer, POE/POP alkyl ether, polyether-modified silicone, lauric acid alkanolamides, alkylamine oxides, hydrogenated soybean phospholipids, and the like.

Examples of the anionic surfactant include fatty acid soap, alpha-acyl sulfonate, alkyl sulfonate, alkyl allyl sulfonate, alkyl naphthalene sulfonate, alkyl sulfate, POE alkyl ether sulfate, alkyl amide sulfate, alkyl phosphate, POE alkyl phosphate, and alkyl amide phosphate. , alkyloylalkyltaurine salts, N-acylamino acid salts, POE alkylethercarboxylate salts, alkylsulfosuccinate salts, alkylsulfoacetate sodium, acylated hydrolyzed collagen peptide salts, perfluoroalkylphosphate esters, and the like. .

Examples of the cationic surfactant include alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, chloride Benzalkonium, diethylaminoethylamide stearate, dimethylaminopropylamide stearate, quaternary ammonium salt of lanolin derivative, etc. are mentioned.

Examples of the amphoteric surfactant include carboxybetaine type, amidebetaine type, sulfobetaine type, hydroxysulfobetaine type, amidesulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, amidamine type, etc. Amphoteric surfactant etc. are mentioned.

Examples of organic and inorganic pigments include silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, bengala, clay, bentonite, titanium-coated mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, aluminum oxide. inorganic pigments such as calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine blue, chromium oxide, chromium hydroxide, calamine, and complexes thereof; Polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenol resin, fluororesin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene/styrene copolymer, and organic pigments such as silk powder, cellulose, CI pigment yellow and CI pigment orange, and composite pigments of these inorganic pigments and organic pigments.

Examples of the organic powder include metal soaps such as calcium stearate; alkyl phosphate metal salts such as sodium cetylate, zinc laurylate, and calcium laurylate; acylamino acid polyvalent metal salts such as calcium N-lauroyl-beta-alanine, zinc N-lauroyl-beta-alanine, and calcium N-lauroyl glycine; amidesulfonic acid polyvalent metal salts such as calcium N-lauroyl-taurine and calcium N-palmitoyl-taurine; N such as N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoylizine, N-alpha-paritoylolnithine, N-alpha-lauroylarginine, N-alpha-hydrogenated beef fatty acid acylarginine, etc. -Acyl basic amino acid; N-acyl polypeptides, such as N-lauroyl glycyl glycine; alpha-amino fatty acids such as alpha-aminocaprylic acid and alpha-aminolauric acid; Polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, a divinylbenzene-styrene copolymer, ethylene tetrafluoride, etc. are mentioned.

As UV absorbers, para-aminobenzoic acid, para-aminobenzoate ethyl, para-aminobenzoate amyl, para-aminobenzoate octyl, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butylphenyl salicylate, homomentyl salicylate, benzyl cinnamate , para-methoxycinnamic acid-2-ethoxyethyl, para-methoxycinnamic acid octyl, dipara-methoxycinnamic acid mono-2-ethylhexaneglyceryl, para-methoxycinnamic acid isopropyl, diisopropyl diisopropyl cinnamic acid ester mixture, uro Cannic acid, ethyl urokanate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenonesulfonic acid and salts thereof, dihydroxymethoxybenzophenone, sodium dihydroxymethoxybenzophenonedisulfonate, dihydroxybenzophenone , tetrahydroxybenzophenone, 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-trianilino-p-(carbo-2'-ethylhexyl-1'-oxy)-1 ,3,5-triazine, 2-(2-hydroxy-5-methylphenyl)benzotriazole, etc. are mentioned.

Examples of disinfectants include hinokitiol, triclosan, trichlorohydroxydiphenyl ether, chlorhexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, zinc phyllithione, benzalkonium chloride, photosensitizer. So, No. 301, mononitroguaial sodium, undecyrenic acid, etc. are mentioned.

Examples of the antioxidant include butylhydroxyanisole, propyl gallic acid, and ellisorbic acid.

Examples of the pH adjuster include citric acid, sodium citrate, malic acid, sodium malate, fmalic acid, sodium fumarate, succinic acid, sodium succinate, sodium hydroxide, sodium monohydrogen phosphate, and the like.

As alcohol, higher alcohols, such as cetyl alcohol, are mentioned.

In addition, blending components that may be added are not limited thereto, and any of the above components can be blended within a range that does not impair the object and effect of the present invention, but preferably 0.01-5% by weight relative to the total weight, More preferably, it is blended in 0.01-3% weight percentage.

When the formulation of the present invention is a lotion, paste, cream or gel, animal fiber, vegetable fiber, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide, etc. can be used

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 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, solvating agent or emulsifying agent is used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylglycol oil, glycerol fatty esters, fatty acid esters of polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, as a carrier component, water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or tracanth may 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, methyl taurate, sarcosinate, fatty acid amide as carrier components Ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linolin derivative or ethoxylated glycerol fatty acid ester and the like may be used.

Hereinafter, preferred examples are presented to help the understanding of the present invention. However, the following examples are only provided for easier understanding of the present invention, and the contents of the present invention are not limited by the following examples.

Example 1. Preparation of Gaeso Sirangaebi Ethanol Extract (Ps-EE)

Potentilla supina was washed with tap water to remove impurities, sterilized using 70% (v/v) ethanol, washed with distilled water, and freeze-dried. The freeze-dried dog sagebrush was finely crushed into a powder, and 1 liter of 80% (v/v) ethanol was added to 100 g of the crushed dog sagebrush and immersed at room temperature for 24 hours. After repeating this process three times, the obtained extract was filtered with Whatman filter paper, and then the solvent was evaporated using an evaporator to obtain an ethanol extract from Gaeso Shirangae.

Example 2. Cell culture method

B16F10 cells, a murine melanoma cell, and HaCaT cells, a human keratinocyte cell, were cultured in DMEM (Dulbecco's modified Eagle's medium) culture medium containing 10% fetal bovine serum (FBS) and penicillin-streptomycin. 37 ℃, 5% carbon dioxide (CO ₂ ) using a cell culture incubator conditions.

Example 3. Confirmation of antioxidant effect according to the inhibition of active oxygen

B16F10 cells were treated with α-MSH and then cultured using an incubator at 5% CO ₂ , 37° C. for 36 hours. Then, H2DCF-DA staining method was used to measure the amount of ROS. It was confirmed whether or not the production and inhibition of reactive oxygen species in the cells. Specifically, the cells were fixed using 300 μl of 5% formaldehyde dissolved in PBS, and 500 μl of 2 mM H2DCFDA (2',7'-dichlorofluorescein diacetate) was added to stain the cells, and 300 μl of 1 μg DAPI solution was added. After the nuclei were stained, the amount of reactive oxygen species (ROS) was analyzed with a fluorescence microscope.

As a result, as shown in FIG. 1 , it was confirmed that the amount of ROS raised by α-MSH was significantly reduced by the ethanol extract of Gaeso Shirangaebi. Based on this, it was confirmed that the ethanol extract of Gaeso Shirangaebi could regulate melanogenesis by regulating ROS in B16F10 cells.

Example 4. Confirmation of cell viability

To measure the toxicity of Ps-EE to cells in the steady state, HaCaT and B16F10 cells were plated at 1 × 10 ⁴ cells/ml in 96-well plates, and 25, 50, 100, 200, 400 μg/ml ml of Ps-EE was treated and cultured. Next, using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphinyltetrazolium bromide) assay, the cell viability of the extract-treated cells as described above was confirmed. Specifically, 10 μl MTT solution (stock concentration: 5 mg/ml) was added to the cultured cells after the extract treatment in the above, and an additional reaction was induced for 3 hours. 100 μl MTT stopping solution (10% Sodium dodecyl sulfate in 0.01M HCl) was additionally added to each well for completion of the reaction and dissolution of formazan crystals. Cell viability was calculated using the OD value obtained by measuring the absorbance at 570 nm in the amount of MTT reduced to promazan.

As shown in FIGS. 2A and 2B , it was confirmed that the cell viability was maintained at 95% or more in HaCaT and B16F10 cells even when the ethanol extract was treated up to 100 μg/ml. Based on this, it can be seen that the ethanol extract of Gaesosirangaebi does not show toxicity to cells.

Example 5. Confirmation of effect on melanin formation and secretion

Example 5-1. Confirmation of effect on melanin secretion

Melanocyte stimulating hormone (α-MSH), a hormone that promotes melanin synthesis, was used to induce melanin formation in melanocytes. The B16F10 cells cultured in Example 2 were aliquoted in a 6-well plate so that 5×10 ⁴ cells per well were cultured for 24 hours, and then treated with an ethanol extract of Gaeso sirangaebi and α-MSH to become 100 nM. The treated cells were cultured for 48 hours in a cell incubator at 37° C., 5% carbon dioxide (CO ₂ ) condition. After 48 hours, 800 μl of the cell culture solution was transferred to a new tube and centrifuged at 12,000 rpm at 4° C. for 5 minutes to obtain a supernatant. 100 μl of the obtained supernatant was dispensed in a 96-well plate and absorbance was measured at 475 nm. At this time, the melanin secretion amount was calculated based on 100% of the melanin secretion amount in the cell culture medium not treated with α-MSH.

As shown in Figure 3a, it was confirmed that the amount of melanin secretion increased by the ethanol extract treatment of Gaesosirangaebi was reduced.

Example 5-2. Confirmation of effect on melanin formation

Melanocyte stimulating hormone (α-MSH), a hormone that promotes melanin synthesis, was used to induce melanin formation in melanocytes. The B16F10 cells cultured in Example 2 were aliquoted in a 6-well plate so that 5×10 ⁴ cells per well were cultured for 24 hours, and then treated with an ethanol extract of Gaeso sirangaebi and α-MSH to become 100 nM. The treated cells were cultured for 48 hours in a cell incubator at 37° C., 5% carbon dioxide (CO ₂ ) condition. After 48 hours, the cell culture solution was removed, and 200 μl of cell lysis buffer was applied to each well to lyse the cells, and then centrifuged at 4° C. at 12,000 rpm for 5 minutes, and then the supernatant was removed. To the obtained precipitate, 100 μl of 10% sodium dodecyl sulfate-1M NaOH solution was dispensed and heated at 60° C. for 30 minutes. After that, the heated solution was left at room temperature, cooled to 25 °C, and absorbance was measured at 405 nm. At this time, the amount of melanin was calculated based on 100% of melanin formation in cells not treated with α-MSH.

As shown in Figure 3b, it was confirmed that the amount of melanin formation, which was increased by the ethanol extract treatment of Gaeso Sirangaebi, was reduced.

Example 6. Confirmation of mRNA expression level of whitening-related factors using RT-PCR

The B16F10 cells cultured in Example 2 were aliquoted in a 6-well plate so that 5 × 10 ⁴ cells per well were cultured for 24 hours, and then treated with ethanol extract and α-MSH so as to become 100 nM cells. was cultured for 48 hours in a cell incubator at 37 ° C., 5% carbon dioxide (CO ₂ ) condition. After 48 hours, all of the cell culture medium was removed, the cells were decomposed using TRI-Zol, and neutralized using BCP. mRNA was precipitated in the neutralized liquid using isopropanol. Then, using a centrifuge, only the precipitated mRNA was collected and used for the experiment. Intracellular mRNA was synthesized into cDNA using a cDNA kit, and the expression levels of Tyrosinase, TYRP-1, TYRP-2, and MITF genes, which are genes important for melanin formation, were confirmed through PCR.

As shown in Figure 4, it was confirmed that the ethanol extract of Gaeso sirangaebi can inhibit melanin formation by down-regulating Tyrosinase, TYRP-1, TYRP-2, and MITF related to melanin formation.

primer order sequence(5'->3') MITF Forward CTGATGGACGATGCCCTCTC Reverse TCCGTTTCTTCTGCGCTCAT Tyrosinase Forward GTCCACTCACAGGGATAGCAG Reverse AGAGTCTCTGTTATGGCCGA TYRP-1 Forward ATGGAACGGGAGGACAAACC Reverse TCCTGACCTGGCCATTGAAC TYRP-2 Forward CAGTTTCCCCGAGTCTGCAT Reverse GTCTAAGGCGCCCAAGAACT HAS-1 Forward CAGCCTGCGATACTGGGTAG Reverse GGCTGGGTCAAGCATAGTGT HAS-2 Forward GGGGGAGATGTCCAGATTTT Reverse ATGCACTGAACACACCCAAA HAS-3 Forward TATACCGCGCGCTCCAA Reverse GCCACTCCCGGAAGTAAGACT Hyal-1 Forward CAGAATGCCAGCCTGATTGC Reverse CCGGTGTAGTTGGGGCTTAG Hyal-2 Forward TACACCACAAGCACGGAGAC Reverse ATGCAGGAAGGTACTGGCAC Hyal-3 Forward CCAGGATGACCTGTGCAGT Reverse CCATCTGTCCTGGATCTCGC Hyal-4 Forward TGAGCTCTCTTGGCTCTGGA Reverse AGGCAGCACTTTCTCCTATGG GAPDH(M) Forward GGGTCCCAGCTTAGGTTCATC Reverse TACGGCCAAATCCGTTCACA GAPDH(H) Forward GACAGTCAGCCGCATCTTCT Reverse GCGCCCAATACGACCAAATC

Example 7. Confirmation of mRNA expression level of moisturizing factors using RT-PCR

The HaCaT cells cultured in Example 2 were aliquoted to 5 × 10 ⁴ cells per well in a 6-well plate, cultured for 24 hours, and then treated with an ethanol extract of Gaeso Shirangaebi at a concentration of 50-100 ug/ml Cells was cultured for 24 hours in a cell incubator at 37 ° C., 5% carbon dioxide (CO ₂ ) conditions. After 24 hours, all of the cell culture medium was removed, the cells were decomposed using TRI-Zol, and neutralized using BCP. mRNA was precipitated in the neutralized liquid using isopropanol. Then, using a centrifuge, only the precipitated mRNA was collected and used for the experiment. Intracellular mRNA is synthesized into cDNA using a cDNA kit, and through PCR, HAS-1, HAS-2, HAS-3, which are important genes for moisturizing, and Hyal-1, Hyal-2, Hyal-, which are genes related to moisturizing factor degradation. 3, the expression level of the Hyal-4 gene was confirmed.

As shown in FIG. 5 , it was confirmed that the ethanol extract of Gaeso Sirangaebi exhibits a moisturizing effect by up-regulating HAS-1, HAS-2, and HAS-3, which are genes important for moisturizing.

In addition, as shown in FIG. 6 , it was found that the ethanol extract of Gaeso Shirangaebi suppresses the decomposition of the moisturizing factor by down-regulating Hyal-1, Hyal-2, Hyal-3, and Hyal-4 related to the decomposition of the moisturizing factor, thereby exhibiting a moisturizing effect. Confirmed.

Summarizing the above examples, the present inventors found that the ethanol extract of Gaeso Shirangaebi has no cytotoxicity, has an excellent antioxidant effect, and reduces melanin secretion and formation, as well as the expression of moisturizing factors HAS-1 to HAS-3. It was confirmed that up-regulation and down-regulation of Hyal-1, Hyal-2, Hyal-3, and Hyal-4 related to decomposition of moisturizing factors.

Therefore, it can be seen that the ethanol extract of Gaesosirangaebi can be used as a composition for moisturizing and whitening, in pharmaceuticals, health functional foods and cosmetics, and materials for their development.

The description of the present invention described above is for illustration, and those of ordinary skill in the art to which the present invention pertains can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.

Claims (11)

Contains Gaeso shiranggabi extract as an active ingredient,
A composition for preventing or improving at least one selected from the group consisting of reduced skin moisture content and pigmentation.
According to claim 1,
The composition is a cosmetic composition or a food composition, characterized in that the composition.
According to claim 1,
The extract is water, alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane (hexane), cyclohexane (cyclohexane), petroleum ether (petroleum ether), subcritical fluid, and a composition characterized in that the extract by one or more solvents selected from the group consisting of supercritical fluid.
According to claim 1,
The Gaesosirangae extract is a composition, characterized in that it reduces the expression or activity of MITF (microphthalmia-associated transcription factor).
According to claim 1,
The Gaesosirangae extract is characterized in that it reduces the expression or activity of tyrosinase, the composition.
According to claim 1,
The composition, characterized in that the Gaesosiranggabi extract reduces the expression or activity of TYRP-1 (Tyrosinase-related protein 1) or TYRP-2 (Tyrosinase-related protein 2).
A composition for skin whitening comprising an extract of Gaeso shirangaebi as an active ingredient.
8. The method of claim 7,
The composition is characterized in that one selected from the group consisting of a pharmaceutical composition, a food composition and a cosmetic composition, the composition.
8. The method of claim 7,
The composition is characterized in that preventing, ameliorating or treating a skin pigmentation disease, the composition.
8. The method of claim 7,
The composition is characterized in that it inhibits melanin secretion, the composition.
10. The method of claim 9,
The skin pigmentation disease is characterized in that at least one selected from the group consisting of spots, freckles, black spots, birthmarks, drug-induced pigmentation, post-inflammatory pigmentation, and hyperpigmentation occurring in dermatitis, the composition.

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