KR20230107933A - Compositions for preventing or improving skin damage comprising the extract of plats as an active ingredient - Google Patents

Abstract

The present invention relates to a composition for preventing or alleviating skin damage comprising a plant extract as an active ingredient. According to one aspect, the composition comprising a plant extract inhibits MMP-1 expression and can be effectively used to prevent or alleviate skin damage by increasing the expression of pro-collagen type and skin barrier-related factors.

Description

Compositions for preventing or improving skin damage comprising a plant extract as an active ingredient {Compositions for preventing or improving skin damage comprising the extract of plats as an active ingredient}

It relates to a composition for preventing or improving skin damage using a plant extract as an active ingredient.

As the elderly population increases, skin beauty and antioxidants that can prevent aging, which can be a measure of aging, are emerging as major concerns. Wrinkles of the skin and reduced antioxidant activity in the body due to aging can be used as a measure of health. Skin aging can be divided into intrinsic aging and extrinsic aging according to factors. Intrinsic aging is known to be caused by changes in physiological functions of the skin epidermis and dermis according to age, and extrinsic aging is known to be caused by changes in physiological functions of the skin caused by environments such as air pollution, exposure to ultraviolet rays, and stress. Among these aging mechanisms, oxidative stress induced by ultraviolet rays increases free radicals in the body, and MMP-1 (Matrix Metalloproteinase-1), which decomposes collagen, hyaluronic acid ( It can be explained by damage to the skin epidermis and dermis due to the increased activity of hyaluronidase that decomposes hyaluronic acid.

The stratum corneum of the skin exists in the outermost layer of the skin and is in direct contact with the external environment, so it plays an important barrier function to protect our body from external physical and chemical stress. This barrier function is maintained by epidermal homeostasis. Epidermal homeostasis maintains a continuous skin barrier function by forming a skin barrier called the stratum corneum through terminal differentiation through the growth division and cell migration of keratinocytes in the basal layer (Korean J Food.Sci.Technol.43:458-463, 2011).

As keratinocytes differentiate, they produce two factors that affect moisturization. First, during differentiation of keratinocytes, the cell membrane is replaced by a structure called a cornified envelope. The keratinocyte membrane is a membrane structure in which various structural proteins including loricrin, involucrin, and filaggrin are cross-linked, providing skin protection against the external environment and evaporating moisture within keratinocytes. (Nat. Rev. Mol. Cell Biol. 6(4):328-340, 2005).

In addition, during the differentiation process of keratinocytes, keratinocytes maintain a function as a skin barrier while producing Natural Moisturizing Factor (NMF). A protein that is an important source for the production of natural moisturizing factors is filaggrin, which is decomposed into hydrophilic amino acids by CASP 14 to form natural moisturizing factors. Natural moisturizing factors function to maintain skin moisturizing power by providing water holding capacity and moisture absorption in the air (J. Cell Sci. 122:1285-1294, 2009). Therefore, maintaining an appropriate level of natural moisturizing factor in the skin is a very important factor for skin health through the skin barrier function.

Therefore, research on mechanisms for preventing or improving skin damage from ultraviolet rays has been actively conducted, and research on cosmetics and health functional foods exhibiting biochemical efficacy and effects is being conducted. Materials with practical efficacy and effects due to the development of the research level and the increase in technology are the main research subjects.

Plant-derived materials have been used for a long time because they are excellent in terms of safety. In particular, in Korea, the development of functional materials mainly made of plants and herbal ingredients used in the private sector or in oriental medicine is being actively conducted.

However, the use of the plant extracts for the prevention or improvement of skin damage is not known, and the mechanism for this has not been studied. Therefore, the present inventors conducted a direct efficacy study on the prevention or improvement of skin damage with the extracts.

Accordingly, the present inventors have tried to develop a composition for preventing or improving skin damage. As a result, the secretion of MMP-1 (Matrix Metalloproteinase-1) is reduced in damaged dermal cells, the secretion of Procollagen type I is increased, and in damaged epidermal cells By increasing the expression of Filaggrin, Involucrin, Loricrin and Caspase14, it was confirmed that skin damage can be prevented or improved, and the present invention was completed.

One aspect is a health functional food composition for preventing or improving skin damage comprising at least one extract selected from the group consisting of extracts of Breea segeta , Buttercup ( Cardamine leucantha ) and Ixeris stolonifera as an active ingredient. is to provide

Another aspect is to provide a cosmetic composition for preventing or improving skin damage, comprising at least one extract selected from the group consisting of extracts of wild parsnip, parsley, and crickets, as an active ingredient.

Another aspect is to provide a pharmaceutical composition for preventing or treating skin damage comprising at least one extract selected from the group consisting of extracts of wild parsnip, parsley, and crickets, as an active ingredient.

Another aspect is to provide a quasi-drug composition for preventing or improving skin damage, comprising, as an active ingredient, one or more extracts selected from the group consisting of extracts of wild parsnip, water parsley, and silverfish.

One aspect is a health functional food composition for preventing or improving skin damage comprising at least one extract selected from the group consisting of extracts of Breea segeta , Buttercup ( Cardamine leucantha ) and Ixeris stolonifera as an active ingredient. provides

The plant is a perennial plant of the Asteraceae family, and may be a plant specified as an edible part in the food raw materials notified by the Ministry of Food and Drug Safety.

The buttercup is a perennial plant of the Cruciferae family, and may be a plant whose leaves are specified as edible parts in food raw materials notified by the Ministry of Food and Drug Safety.

The silverfish is a perennial plant of the Asteraceae family, and may be a plant whose roots and leaves are specified as edible parts in food raw materials notified by the Ministry of Food and Drug Safety.

In one embodiment, the extract may be extracted from at least one selected from the group consisting of the whole plant, root, stem, branch, leaf, seed and fruit of the extracts of A. Whole, part of, or materials derived therefrom of the extracts of the wild parsnip, parsley, and silverfish used in the extraction may be ground or shredded or suitably dried.

An extraction method for obtaining the extract may be prepared according to a known extraction method commonly used in the art to which the present invention belongs. Specifically, commonly used extraction methods such as cold precipitation, heat extraction, ultrasonic extraction, filtration, pressure extraction, reflux extraction, supercritical extraction, and electrical extraction may be used, and a conventional extraction device, ultrasonic crushing extractor or fractionator may be used. .

In addition, if necessary, after the above extraction, filtration and/or concentration and/or freeze-drying methods known in the art may be additionally used. The dried or concentrated extract used in the present invention means dried or concentrated by a known method as described above.

In one embodiment, the extract may be extracted using water, alcohol, or a mixture thereof as a solvent. The alcohol may be a C1 to C6 alcohol, for example a C1 to C4 alcohol. The C1 to C6 alcohol may be methanol, ethanol, propanol, isopropanol, 1,3-propanediol, butanol, pentanol or hexanol.

In one embodiment, the extraction is 3 to 10 (volume or weight) times the extraction solvent for each extract, for example, 3 to 7 (volume / weight) times, 3 to 5 (volume / weight) times , 5 to 10 (volume / weight) times, or 4 to 10 (volume / weight) times.

In one embodiment, the solvent may be a mixture of water and alcohol, that is, an aqueous alcohol solution, for example, an aqueous ethanol solution. The alcohol concentration in the alcohol aqueous solution is 1 to 99.5 (v / v)%, 10 to 99.5 (v / v)%, 1 to 70 (v / v)%, 1 to 40 (v / v)%, 5 to 25 (v/v)%, 7 to 20 (v/v)%, 5 to 25 (v/v)%, or 10 to 20 (v/v)%.

In one embodiment, the extract increases the expression or activity of any one or more selected from the group consisting of Filaggrin, Involucrin, Loricrin and Caspase 14 can

In one embodiment, the extract may inhibit the expression or activity of MMP-1.

In one embodiment, the extract may be to increase the expression or activity of procollagen type I (Procollagen type I).

The present inventors found that the extract is effective in moisturizing the skin by increasing the expression or activity of Filaggrin, Involucrin, Loricrin and/or Caspase 14, and furthermore, it causes skin damage. It has been confirmed that it can be prevented, ameliorated or treated. In addition, it was confirmed that skin damage can be prevented, improved or treated by increasing the expression or activity of procollagen type I and inhibiting the expression or activity of MMP-1.

As used herein, the term "skin damage" includes external stimuli, for example, death of human skin cells by ultraviolet rays, skin cell DNA damage, increased reactive oxygen species, increased lipid peroxidation, etc., the symptoms of which include erythema, It may include sunburn, hyperpigmentation, photoaging, skin cancer, and the like.

In one embodiment, the skin damage may be caused by ultraviolet rays.

As used herein, the term “prevention” refers to partially or completely delaying or preventing the onset or recurrence of a disease, disorder, or its attendant symptoms, preventing the acquisition or re-acquisition of a disease or disorder, or preventing the occurrence or recurrence of a disease or disorder. Reducing the risk of acquisition. The prevention refers to any action that inhibits or delays the occurrence of a skin damaging disease, disorder, or symptom by administering the composition according to the present invention.

As used herein, the term "improvement" refers to any action that at least reduces a parameter associated with the condition being treated, eg, the severity of a symptom.

In one embodiment, the extract is 0.0001% to 99.0% by weight relative to the total weight of the composition, for example, 0.01% to 60% by weight, 0.01% to 40% by weight, 0.01% to 30% by weight, 0.01% to 30% by weight 0.01 wt% to 10 wt%, 0.01 wt% to 5 wt%, 0.05 wt% to 60 wt%, 0.05 wt% to 40 wt%, 0.05 wt% to 30 wt%, 0.05 wt% to 20% by weight, 0.05% to 10% by weight, 0.05% to 5% by weight, 0.1% to 60% by weight, 0.1% to 40% by weight, 0.1% to 30% by weight, 0.1% to 20% by weight % by weight, 0.1% to 10% by weight, 0.1% to 5% by weight, 1% to 50% by weight, 1% to 40% by weight, 2% to 40% by weight, 3% to 30% by weight 4% to 30%, 5% to 30%, 5% to 20%, 5% to 18%, 6% to 15%, 8% to 15% or 7% % to 10% by weight may be included. However, in the case of long-term intake for the purpose of health and hygiene or health control, it may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

In this specification, the term "health functional food" refers to a food manufactured and processed by using a specific ingredient as a raw material or by extracting, concentrating, refining, mixing, etc. a specific ingredient contained in a food raw material for the purpose of health supplement. It refers to food designed and processed so that it can sufficiently exert biological control functions such as biological defense, regulation of biological rhythm, prevention and recovery of disease, etc. The health functional food composition may perform functions related to prevention and improvement of skin damage caused by fine dust.

There is no particular limitation on the type of food. Examples of foods to which the extract may be added include powders, granules, tablets, capsules, pills, gels, jellies, suspensions, emulsions, syrups, tea bags, leached teas, gums, candies, and formulations selected from the group consisting of health drinks etc., and includes all health foods in the usual sense.

The health beverage composition may include various sweeteners, flavoring agents, or natural carbohydrates as additional components, as in conventional beverages. The sweetener may be a natural sweetener or a synthetic sweetener. The natural sweetener may be taumatin or stevia extract, and the synthetic sweetener may be saccharin or aspartame.

The natural carbohydrate may be monosaccharide, disaccharide, polysaccharide, xylitol, sorbitol or erythritol. The monosaccharide may be glucose or fructose, and the disaccharide may be maltose or sucrose. The polysaccharide may be a dextrin or a cyclodextrin. The proportion of the natural carbohydrate may be generally about 0.01 to 10 g, for example, about 0.01 to 0.1 g per 100 ml of the composition of the present invention.

The health functional food may include food additives acceptable in food science, and may include appropriate carriers commonly used in the manufacture of health functional food.

In addition to the above, the composition of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages; and the like. In addition, the composition of the present invention may include fruit flesh for preparing natural fruit juice, fruit juice beverages, and vegetable beverages. These components may be used independently or in combination. The ratio of these additives is not critical, but is generally selected in the range of 0.01 to 0.1 part by weight per 100 parts by weight of the composition of the present invention.

Another aspect provides a cosmetic composition for preventing or improving skin damage, comprising at least one extract selected from the group consisting of extracts of wild parsnip, buttercup, and cactus extract as an active ingredient.

The cosmetic composition includes lotion (skin lotion), toner, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrient lotion, massage cream, nutrient cream, moisture cream, hand cream, hand sanitizer, foundation, essence, Nutritional essence, packs, soaps, cleansing foams, cleansing lotions, cleansing creams, body lotions, body cleansers, suspensions, gels, powders, pastes, mask packs, and can be prepared into formulations including sheets. Compositions of such formulations may be prepared according to conventional methods in the art. The blending amount of the additional ingredients such as the moisturizing agent can be easily selected by those skilled in the art within a range that does not impair the objects and effects of the present invention.

The cosmetic composition may further include functional additives and ingredients included in general cosmetic compositions in addition to the active ingredients disclosed herein, and commonly used purified water, thickeners, preservatives, stabilizers, solubilizers, surfactants, carriers, A flavoring agent or a combination thereof may be further included. The functional additive may include a component selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high-molecular peptides, high-molecular polysaccharides, sphingolipids, and seaweed extracts. Alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, humectants, viscosity modifiers, emulsions, stabilizers, UV scattering agents, UV absorbers, coloring agents, perfumes, and the like may be exemplified as the carrier. Compounds/compositions that can be used as the alcohol, oil, surfactant, fatty acid, silicone oil, wetting agent, humectant, viscosity modifier, emulsion, stabilizer, UV scattering agent, UV absorber, coloring agent, fragrance, etc. are already known in the art. Since there is, those skilled in the art can select and use an appropriate corresponding material / composition. In addition, the cosmetic composition, if necessary, sunscreen, antioxidants (butylhydroxyanisole, gallic acid propyl, ellisorbic acid, tocopheryl acetate, butylated hydroxytoluene, etc.), preservatives (methylparaben, butylparaben , propylparaben, phenoxyethanol, imidazolidinylurea, chlorphenesin, etc.), colorant, pH adjuster (triethanolamine, citric acid, citric acid, sodium citrate, malic acid, sodium malate, fumaric acid, sodium fumarate, succinic acid , sodium succinate, sodium hydroxide, sodium hydrogen phosphate, etc.), humectants (glycerin, sorbitol, propylene glycol, butylene glycol, hexylene glycol, diglycerin, betaine, glycereth-26, methylglue Seth-20, etc.) and lubricants may be further added.

In addition, in the cosmetic composition of each formulation, appropriate components may be selected and blended according to the cosmetic formulation or purpose of use. Since the formulation components and methods may follow conventional techniques, detailed descriptions thereof are omitted herein.

Another aspect provides a pharmaceutical composition for preventing or treating skin damage comprising, as an active ingredient, at least one extract selected from the group consisting of extracts of wild parsnip, parsley, and cactus.

As used herein, the term "pharmaceutical composition" can refer to a molecule or compound that imparts some beneficial effect upon administration to a subject. Beneficial effects may include enabling diagnostic determination; amelioration of a disease, symptom, disorder or condition; reducing or preventing the occurrence of a disease, condition, disorder or condition; and responding to a disease, symptom, disorder or condition in general.

As used herein, the term "treatment" refers to any action that improves or beneficially alters a disease, disorder, or its attendant symptoms.

The pharmaceutical composition may be formulated in a formulation selected from the group consisting of tablets, soft or hard capsules, pills, powders, suspensions, syrups, injections and granules.

The pharmaceutical composition may be for oral or parenteral administration.

The pharmaceutical composition includes conventional fillers, extenders, binders, disintegrants, anti-agglomerates, lubricants, wetting agents, pH adjusters, nutrients, vitamins, electrolytes, alginic acid and its salts, pectic acid and its salts, protective chlorides, glycerin, Perfumes, emulsifiers or preservatives may be included.

The pharmaceutical composition may include a pharmaceutically acceptable carrier, examples of which include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, and gelatin. , calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, propylhydroxybenzoate, It may be at least one selected from the group consisting of talc, magnesium stearate and mineral oil, dextrin, calcium carbonate, propylene glycol, liquid paraffin and physiological saline.

The formulation of the pharmaceutical composition may vary depending on the method of use, and is formulated using a method well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. It can be.

Formulations for oral administration include tablets, soft or hard capsules, pills, powders, suspensions, syrups, injections and granules, etc., and these formulations include one or more excipients such as starch, calcium carbonate, sucrose or lactose. , It may be prepared by mixing gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Formulations for parenteral administration may be creams, lotions, ointments, warnings, liquids, aerosols, liquid extracts, elixirs, salivas, sachets, patches, or injections.

The method is applicable to any animal, and the animal may include livestock such as cattle, pigs, sheep, horses, dogs, and cats, as well as humans and primates.

The dosage of the composition for treatment, prevention, or improvement may be determined in consideration of the administration method, the age, sex, severity of the patient, the degree of absorption of the active ingredient in the body, the inactivation rate, and concomitant drugs, 0.1 mg/kg (body weight) to 500 mg/kg (body weight), 0.1 mg/kg (body weight) to 400 mg/kg (body weight), or 1 mg/kg (body weight) to 300 based on the daily active ingredient It may be administered in mg/kg (body weight), and may be administered once or divided into several times, but is not limited thereto.

Another aspect provides a quasi-drug composition for preventing or improving skin damage, comprising, as an active ingredient, one or more extracts selected from the group consisting of extracts of wild parsnip, water parsley, and silverfish.

The term “quasi-drugs” refers to textiles, rubber products, or similar products used for the purpose of treating, mitigating, treating or preventing human or animal diseases, weak or non-directly acting on the human body, and not instruments or machines. Products used for the purpose of diagnosing, treating, mitigating, treating, or preventing human or animal conditions or diseases, which fall under one of the following categories: It refers to items other than instruments, machines, or devices, and items other than instruments, machines, or devices among items used for the purpose of pharmacologically affecting the structure and function of humans or animals, including external skin products and personal hygiene products. can include

When the extract is added to a quasi-drug composition, the extract may be added as it is or used together with other quasi-drug ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of use (prevention, health or therapeutic treatment).

The quasi-drug composition is not particularly limited thereto, but may be a personal hygiene product, an external skin preparation, a humidifier filler, a mask, an ointment, or a filter filler. The personal hygiene product may be soap, wet tissue, toilet paper, shampoo, toothpaste, hair care product, air freshener gel or cleansing gel.

A composition containing a plant extract according to one aspect inhibits MMP-1 expression and increases the expression of procollagen type I and skin barrier-related factors, thereby effectively preventing or improving skin damage.

Figure 1 is a graph showing the cell viability according to treatment with extracts of chamomile, parsley, or cactus in dermal cells damaged by UV-B:
Figure 1 (a) is a graph showing the cell viability according to the treatment of each concentration of the extract of chabaengi extract; Figure 1 (b) is a graph showing the cell viability according to the concentration of watercress extract; And Figure 1 (c) is a graph showing the cell viability according to the treatment of each concentration of the extract of the cilia extract.
Figure 2 is a graph showing the cell viability according to the treatment of the extracts of sagebrush, buttercup, or cactus in epidermal cells damaged by UV-B:
Figure 2 (a) is a graph showing the cell viability according to the treatment of each concentration of the extract of chabaengi extract; Figure 2 (b) is a graph showing the cell viability according to the concentration of watercress extract; And Figure 2 (c) is a graph showing the cell viability according to the treatment of each concentration of the extract of the c.
Figure 3 is a graph showing the amount of MMP-1 secretion according to the treatment of the dermal cells damaged by UV-B;
Figure 3 (a) is a graph showing the amount of secretion of MMP-1 according to the treatment of each concentration of the extract of chabaengi; Figure 3 (b) is a graph showing the amount of secretion of MMP-1 according to the concentration of the buttercup extract; and FIG. 3(c) is a graph showing the secretion amount of MMP-1 according to the treatment of each concentration of the c.
Figure 4 is a graph showing the amount of secretion of Procollagen type I according to the treatment of extracts of C. japonica, Parsley parsnips, or Raccoonsis in dermal cells damaged by UV-B:
Figure 4 (a) is a graph showing the amount of secretion of Procollagen type I according to the treatment of each concentration of the extract of chabaengi; Figure 4 (b) is a graph showing the secretion amount of Procollagen type I according to the concentration of the buttercup extract; and FIG. 4(c) is a graph showing the amount of secretion of Procollagen type I according to the treatment of each concentration of the c.
5 is a graph showing the expression levels of Filaggrin, Involucrin, Loricrin, and Caspase14 according to the treatment of the extract of Chabaengi in epidermal cells damaged by UV-B.

It will be described in more detail through the following examples. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

Example 1. Preparation of extracts of wild parsnip, water parsley and silverfish

In order to manufacture extracts of wild parsnips, water parsley, and cicadas, plants that grow wild in Korea are purchased, washed with distilled water, and thoroughly washed in a cool place away from sunlight until there is no change in weight. dried.

Then, the dried product completely dried was pulverized using a homogenizer, and 30 g of the pulverized product was added to 900 g of an extraction solvent in which ethanol and distilled water were mixed at a weight ratio of 3:7. At this time, the mixing ratio of the pulverized material and the extraction solvent was set to 30 times the weight ratio. Next, the extraction solvent mixed with the pulverized product was placed in a shaking water bath and extracted with stirring at 60° C. for 5 hours. Then, insoluble substances were removed from the material obtained by extraction using Whatman (No. 2) extraction solvent filter paper at room temperature.

Next, the extracts were concentrated under reduced pressure in a distillation apparatus equipped with a cooling condenser to completely remove the extraction solvent, thereby obtaining extracts of A.

Experimental Example 1. Evaluation of cytotoxicity in dermal cells of extracts of Crayfish, Buttercup, and G.

The cytotoxicity of the extracts of Cabbage japonica, Parsley japonica extract, and Raccoonsis extract of Example 1 was confirmed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay.

Specifically, dermal cells, Hs68 fibroblasts, were cultured in DMEM medium containing 10% FBS and 1% antibiotic-antimycotic at 5% CO ₂ and 37°C. And Hs68 fibroblast cells were stabilized in a 24-well plate at a concentration of 8.0 Х 10 ⁴ cells/500 μl/well for 24 hours. Then, in order to irradiate UV-B, the existing medium was removed, PBS (Phosphate Buffer Solution) was washed, and UV-B was treated with 20 mJ/cm ² . And each extract of Example 1 was treated at a concentration of 12.5, 25, 50 or 100 ppm and cultured for 24 hours. After culturing for 24 hours, cell viability was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. More specifically, each cell was treated with MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) solution to reach a final concentration of 0.5 mg/ml and incubated for 4 hours. After that, the solution was completely removed and the absorbance of the plate dissolved in DMSO (Dimethyl sulfoxide) was measured at 590 nm.

Figure 1 is a graph showing the cell viability according to the treatment of a parsley extract, a parsley parsley extract, or a cactus extract in dermal cells damaged by UV-B.

Figure 1 (a) is a graph showing the cell viability according to the treatment of each concentration of the extract of chabaengi extract; Figure 1 (b) is a graph showing the cell viability according to the treatment of each concentration of the water parsley extract; And Figure 1 (c) is a graph showing the cell viability according to the treatment of each concentration of the extract of the c.

As shown in FIG. 1, the extracts of Cabbage, Buttercup, or Raccoons did not show cytotoxicity, and the treatment concentrations of the experimental examples were determined based on the cytotoxicity test.

Experimental Example 2. Evaluation of Cytotoxicity in Epidermal Cells of Extracts of Cabbage, Buttercup, and Raccoonalis

The cytotoxicity of the extracts of Cabbage japonica, Parsley japonica extract, and Raccoonsis extract of Example 1 was confirmed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay.

Specifically, Hacat epidermal cells were cultured in DMEM medium containing 10% FBS and 1% antibiotic-antimycotic at 5% CO ₂ and 37°C. Hacat cells were then stabilized in a 96-well plate at a concentration of 1.0 Х 10 ⁴ cells/200 μl/well for 24 hours. Then, in order to irradiate UV-B, the existing medium was removed, PBS (Phosphate Buffer Solution) was washed, and UV-B was treated at 18 mJ/cm ² . And each extract of Example 1 was treated at a concentration of 12.5, 25, 50 or 100 ppm and cultured for 24 hours. After culturing for 24 hours, cell viability was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. More specifically, each cell was treated with MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) solution to reach a final concentration of 0.5 mg/ml and incubated for 4 hours. After that, the solution was completely removed and the absorbance of the plate dissolved in DMSO (Dimethyl sulfoxide) was measured at 590 nm.

Figure 2 is a graph showing the cell viability according to the treatment of the extract, buttercup extract, or cactus extract in epidermal cells damaged by UV-B.

Figure 2 (a) is a graph showing the cell viability according to the treatment of each concentration of the extract of chabaengi extract; Figure 2 (b) is a graph showing the cell viability according to the treatment of each concentration of the water parsley extract; And Figure 2 (c) is a graph showing the cell viability according to the treatment of each concentration of the extract of the c.

As shown in FIG. 2, the extracts of Cabbage, Buttercup, or Little Beetle did not show cytotoxicity, and based on the cytotoxicity test, the treatment concentration of the following experimental example was determined.

Experimental Example 3. Confirmation of MMP-1 (Matrix Metalloproteinase-1) Expression Inhibition of Chobaengi, Water Buttercup, and Little Swallowtail Extracts

In order to confirm the efficacy of inhibiting the secretion of MMP-1 on cells damaged by UV-B of the extract of Example 1, the extract of Buttercup, and the extract of A. , abcam, US).

Specifically, dermal cells, Hs68 fibroblasts, were cultured in DMEM medium containing 10% FBS and 1% antibiotic-antimycotic at 5% CO ₂ and 37°C. And Hs68 fibroblast cells were stabilized in a 24-well plate at a concentration of 8.0 Х 10 ⁴ cells/500 μl/well for 24 hours. Then, in order to irradiate UV-B, the existing medium was removed, PBS (Phosphate Buffer Solution) was washed, and UV-B was treated with 20 mJ/cm ² . And each extract of Example 1 was treated at a concentration of 12.5, 25, 50 or 100 ppm and cultured for 24 hours. MMP-1 in the supernatant obtained after 24 hours was quantified using an ELISA kit, and the results are shown in FIG. 3 .

Figure 3 is a graph showing the amount of MMP-1 secretion according to the treatment of an extract, buttercup extract, or a cactus extract in cells damaged by UV-B treatment.

Figure 3 (a) is a graph showing the amount of MMP-1 secretion according to the treatment of each concentration of the extract of chabaengi; Figure 3 (b) is a graph showing the amount of MMP-1 secretion according to the concentration of watercress extract; and FIG. 3(c) is a graph showing the amount of MMP-1 secretion according to the treatment of each concentration of the extract of A.

As shown in FIG. 3, it was confirmed that all of the extracts of Chobaengi, Buttercup, or Raccoonsis significantly inhibited the amount of MMP-1 secretion in proportion to the concentration.

The above results indicate that the extract according to one embodiment effectively inhibits the secretion of MMP-1 and is effective in preventing or improving skin damage caused by ultraviolet rays.

Experimental Example 4. Confirmation of Procollagen type I expression promotion of extracts of Chobaengi, Buttercup, and Ginseng

In order to confirm the effect of increasing the secretion of Procollagen on cells damaged by UV-B of the extract of Example 1, the extract of Buttercup, and the extract of P. It was measured using a kit (Mk101, Takara, japan).

Specifically, dermal cells, Hs68 fibroblasts, were cultured in DMEM medium containing 10% FBS and 1% antibiotic-antimycotic at 5% CO ₂ and 37°C. And Hs68 fibroblast cells were stabilized in a 24-well plate at a concentration of 8.0 Х 10 ⁴ cells/500 μl/well for 24 hours. Then, in order to irradiate UV-B, the existing medium was removed, PBS (Phosphate Buffer Solution) was washed, and UV-B was treated with 20 mJ/cm ² . And each extract of Example 1 was treated at a concentration of 12.5, 25, 50 or 100 ppm and cultured for 24 hours. Procollagen type I in the supernatant obtained after 24 hours was quantified using an ELISA kit, and the results are shown in FIG. 4 .

4 is a graph showing the amount of Procollagen type I secretion according to the treatment of an extract, a parsley parsley, or a cactus extract in cells damaged by UV-B treatment.

Figure 4 (a) is a graph showing the amount of secretion of Procollagen type I according to the treatment of each concentration of the extract of chabaengi; Figure 4 (b) is a graph showing the amount of Procollagen type I secretion according to the concentration of the buttercup radish extract; and FIG. 4(c) is a graph showing the amount of secretion of Procollagen type I according to the treatment of the concentration of the extract of A.

As shown in FIG. 4, it was confirmed that all of the extracts of Jobengi, Buttercup, or Raccoonsis significantly increased the amount of Procollagen type I secretion in proportion to the concentration.

The above result means that in the case of the extract according to one embodiment, it is effective in preventing or improving skin damage caused by ultraviolet rays by effectively increasing the amount of secretion of Procollagen type I.

Experimental Example 5. Evaluation of changes in the expression of genes related to the skin barrier by the extract of C.

In order to confirm the skin moisturizing effect on the cells damaged by UV-B of the extract of Cactus japonica of Example 1, the skin barrier-related genes Filaggrin, Involucrin, Loricrin, and Caspase14 were measured.

Specifically, Hacat epidermal cells were cultured in DMEM medium containing 10% FBS and 1% antibiotic-antimycotic at 5% CO ₂ and 37°C. Hacat cells were then stabilized in a 6-well plate at a concentration of 8.0 Х 10 ⁵ cells/2㎖/well for 24 hours. Then, in order to irradiate UV-B, the existing medium was removed, PBS (Phosphate Buffer Solution) was washed, and UV-B was treated at 18 mJ/cm ² . In addition, the extract of Example 1 was treated at concentrations of 12.5, 25, and 50 ppm and cultured for 24 hours. After 24 hours, mRNA was extracted from the cells, synthesized into cDNA, and quantitative real-time PCR was performed using a target template (primer) to finally evaluate the expression levels of Filaggrin, Involucrin, Loricrin, and Caspase14 genes, and the results were analyzed. 5. The primer sequences used to synthesize cDNA are shown in Table 1.

gene primer direction Primer sequence (5'→3') sequence number Filaggrin Forward AGT GCA CTC AGG GGG CTC ACA One Reverse CCG GCT TGG CCG TAA TGT GT 2 Involucrin Forward TTG GTC AGT GAA GCG ATG AG 3 Reverse AGA TCT GTC TGC AGG GCT GT 4 Loricrin Forward TCA TAA GAA ACC CCG CTG AG 5 Reverse AAG GAA GGA GAG CCT GGA AG 6 Caspase14 Forward CAA ACA CAT GGG TCA CTT GC 7 Reverse CAG AAC TGC TGA GCC TAC CC 8 β-actin Forward CAT GTA CGT TGC TAT CCA GGC 9 Reverse CTC CTT AAT GTC ACG CAC GAT 10

Figure 5 is a graph showing the expression levels of Filaggrin, Involucrin, Loricrin, and Caspase14 derived from UV-B treatment according to the treatment of the extract of Jobengi.

The above results mean that, in the case of saprophytes, the expression levels of Filaggrin, Involucrin, Loricrin, and Caspase14 are effectively increased, which is effective in preventing or improving skin damage caused by ultraviolet rays.

Preparation Example 1. Preparation of tablets

Tablets were prepared by mixing the extracts of Example 1 in the component ratios shown in Table 2 below, and tableting according to a conventional tablet manufacturing method. It was confirmed that the prepared tablets were stable under all test conditions of the formulation examples.

raw material name unit weight (mg) raw material name unit weight (mg) raw material name unit weight (mg) Chobaengi Extract 50 water parsley
extract 50 wormwood extract 50 corn starch 100 corn starch 100 corn starch 100 lactose 100 lactose 100 lactose 100 stearic acid 2 stearic acid 2 stearic acid 2

Production Example 2. Production of capsules

The extracts of Example 1 were mixed in the ingredient ratios shown in Table 3 below, and filled in gelatin capsules to prepare soft capsules. The prepared capsules were confirmed to be stable in all formulation test conditions.

raw material name unit weight (mg) raw material name unit weight (mg) raw material name unit weight (mg) Chobaengi Extract 50 water parsley
extract 50 wormwood extract 50 corn starch 100 corn starch 100 corn starch 100 lactose 100 lactose 100 lactose 100 stearic acid 2 stearic acid 2 stearic acid 2

Preparation Example 3. Preparation of liquid formulation

The extract of Example 1 was mixed in the ingredient ratio shown in Table 4 below, and filled into a bottle or pouch according to a beverage manufacturing method suitable for taste to prepare a liquid formulation. It was confirmed that the prepared liquid preparation was stable under all formulation example test conditions.

raw material name unit weight (g) raw material name unit weight (g) raw material name unit weight (g) Chobaengi Extract 2.5050 Buttercup Extract 2.5050 wormwood extract 2.5050 xanthan gum 0.0075 xanthan gum 0.0075 xanthan gum 0.0075 fructooligosaccharide solution 0.7500 fructooligosaccharide solution 0.7500 fructooligosaccharide solution 0.7500 Coconut Flower Extract Powder 1.0500 Coconut Flower Extract Powder 1.0500 Coconut Flower Extract Powder 1.0500 Ssanghwa Concentrate 1.5000 Ssanghwa Concentrate 1.5000 Ssanghwa Concentrate 1.5000 Red Ginseng Fragrance 0.0450 Red Ginseng Fragrance 0.0450 Red Ginseng Fragrance 0.0450 Purified water 9.1425 Purified water 9.1425 Purified water 9.1425

Preparation Example 4. Preparation of jelly

The extract of Example 1. was mixed in the composition ratio shown in Table 5 below, and the jelly was prepared by filling a three-sided cloth according to a jelly manufacturing method suitable for the taste. The prepared jelly was confirmed to be stable under all formulation test conditions.

raw material name unit weight (g) raw material name unit weight (g) raw material name unit weight (g) Chobaengi Extract 2.0000 Buttercup Extract 2.0000 wormwood extract 2.0000 food gel 0.3600 food gel 0.3600 food gel 0.3600 carrageenan 0.0600 carrageenan 0.0600 carrageenan 0.0600 calcium lactate 0.1000 calcium lactate 0.1000 calcium lactate 0.1000 sodium citrate 0.0600 sodium citrate 0.0600 sodium citrate 0.0600 complex gold extract 0.0200 complex gold extract 0.0200 complex gold extract 0.0200 Enzymatically Treated Stevia 0.0440 Enzymatically Treated Stevia 0.0440 Enzymatically Treated Stevia 0.0440 fructooligosaccharide solution 5.0000 fructooligosaccharide solution 5.0000 fructooligosaccharide solution 5.0000 Red Grape Concentrate 2.4000 Red Grape Concentrate 2.4000 Red Grape Concentrate 2.4000 Purified water 13.9560 Purified water 13.9560 Purified water 13.9560

The composition ratios of Preparation Examples 1 to 4 were generally prepared as formulation examples by mixing suitable ingredients, but the mixing ratio and raw materials may be arbitrarily changed as needed.

Since the extract of the present invention is stable under all formulation test conditions, there was no problem with the stability of the formulation.

<110> COSMAX BIO CO., LTD. <120> Compositions for preventing or improving skin damage comprising the extract of plats as an active ingredient <130> PN220009KR <160> 10 <170> KoPatentIn 3.0 <210> 1 <211> 21 <212> DNA <213> artificial sequence <220> <223> forward primer for Filaggrin <400> 1 agtgcactca gggggctcac a 21 <210> 2 <211> 20 <212> DNA <213> artificial sequence <220> <223> reverse primer for Filaggrin <400> 2 ccggcttggc cgtaatgtgt 20 <210> 3 <211> 20 <212> DNA <213> artificial sequence <220> <223> forward primer for Involucrin <400> 3 ttggtcagtg aagcgatgag 20 <210> 4 <211> 20 <212> DNA <213> artificial sequence <220> <223> reverse primer for Involucrin <400> 4 agatctgtct gcagggctgt 20 <210> 5 <211> 20 <212> DNA <213> artificial sequence <220> <223> forward primer for Loricrin <400> 5 tcataagaaa ccccgctgag 20 <210> 6 <211> 20 <212> DNA <213> artificial sequence <220> <223> reverse primer for Loricrin <400> 6 aaggaaggag agcctggaag 20 <210> 7 <211> 20 <212> DNA <213> artificial sequence <220> <223> forward primer for Caspase14 <400> 7 caaacacatg ggtcacttgc 20 <210> 8 <211> 20 <212> DNA <213> artificial sequence <220> <223> reverse primer for Caspase14 <400> 8 cagaactgct gagcctaccc 20 <210> 9 <211> 21 <212> DNA <213> artificial sequence <220> <223> forward primer for beta-actin <400> 9 catgtacgtt gctatccagg c 21 <210> 10 <211> 21 <212> DNA <213> artificial sequence <220> <223> reverse primer for beta-actin <400> 10 ctccttaatg tcacgcacga t 21

Claims (12)

A health functional food composition for preventing or improving skin damage comprising at least one extract selected from the group consisting of extracts of Breea segeta , Watercress ( Cardamine leucantha ) and Ixeris stolonifera extract as an active ingredient. The method according to claim 1, wherein the extract is extracted with water, alcohol, or a mixed solvent thereof, health functional food composition. The method according to claim 1, wherein the extract increases the expression or activity of any one or more selected from the group consisting of Filaggrin, Involucrin, Loricrin and Caspase 14 , Health functional food composition. The method according to claim 1, wherein the extract is to inhibit the expression or activity of MMP-1, health functional food composition. The method according to claim 1, wherein the extract is to increase the expression or activity of procollagen type I (Procollagen type I), health functional food composition. The health functional food composition according to claim 1, wherein the skin damage is caused by ultraviolet rays. The method according to claim 1, wherein the extract is to be included in 0.0001% to 99.0% by weight relative to the total weight of the composition, health functional food composition. The health functional food composition according to claim 1, wherein the extract is extracted from at least one selected from the group consisting of whole plants, roots, stems, branches, leaves, seeds, and fruits of C. The method according to claim 1, wherein the health functional food is a formulation selected from the group consisting of powders, granules, tablets, capsules, pills, gels, jellies, suspensions, emulsions, syrups, tea bags, leachate, gum, candy and health drinks Having, health functional food composition. A cosmetic composition for preventing or improving skin damage comprising at least one extract selected from the group consisting of extracts of Breea segeta , Buttercup ( Cardamine leucantha ) and Ixeris stolonifera extract as an active ingredient. A pharmaceutical composition for preventing or treating skin damage comprising at least one extract selected from the group consisting of extracts of Breea segeta , Buttercup ( Cardamine leucantha ) and Ixeris stolonifera extract as an active ingredient. A quasi-drug composition for preventing or improving skin damage, comprising at least one extract selected from the group consisting of extracts of Breea segeta , Watercress ( Cardamine leucantha ) and Ixeris stolonifera extract as an active ingredient.