KR20200142397A - Use of Gel Comprising Plant As Raw Material - Google Patents

Abstract

The present invention relates to the use of a gel comprising a plant as a raw material. The plant gel of the present invention can be used to prepare a gel-phase composition using plant powder as a raw material through the same process for various plant types in a batch without an individual extraction and fractionation process according to the plant types as in the conventional plant extract composition. In this regard, the plant gel of the present invention has excellent process efficiency and has usefulness as various cosmetics such as skin moisturizing effect, skin barrier strengthening effect, skin aging prevention effect, UV protection effect, whitening effect and the like depending on the raw material of each plant gel, and thus can be useful as a cosmetic composition.

Description

Use of Gel Comprising Plant As Raw Material {Use of Gel Comprising Plant As Raw Material}

The present invention relates to the use of a gel comprising plants as raw materials.

[Background Art] [0002] Compositions for various uses, including food and beauty applications, made from natural products, particularly plants, have been developed from the prior art. In the case of a plant-based composition, a plant powder obtained by pulverizing the plant raw material was included as a constituent or an extract obtained by extracting an active ingredient from a plant was prepared as a constituent. However, when the plant powder itself is used as a constituent of the cosmetic composition, the dispersibility of the plant powder in the solution of the composition is low, so that a delamination phenomenon in which the solvent and the powder are separated from each other or precipitated in the solution easily occurred. On the other hand, when plant extracts are included as an active ingredient in the composition, the extraction process for extracting the active ingredient is complicated and expensive equipment and cost are required, so it is not economical, and the properties of the active ingredient are different for each type of plant. There was a problem that the equipment and extraction process could not be applied to various plants. Therefore, there is an urgent need for a method for improving dispersibility in a solution for plant powder that can be applied to various plants without going through an extraction process in a method for preparing a composition using plants as raw materials.

Korean Patent Registration No. 10-0854691

In order to solve the above problems, the present inventors have developed a method for preparing a gel-like composition having excellent dispersion stability and feeling of use without adding a separate dispersant or stabilizer when an oxidation process is introduced into the powder of various finely ground plants. It was also confirmed that these vegetable gels have excellent effects as cosmetics such as skin moisturizing effect, anti-aging effect, and whitening effect. Accordingly, an object of the present invention is to provide a plant gel cosmetic composition comprising an oxidized pulverized plant powder and a residual amount of a solvent, and a use thereof.

According to an aspect of the present invention, the present invention provides a plant gel cosmetic composition comprising an oxidized pulverized plant powder and a residual amount of a solvent.

In the present invention, "plant" may be a land plant or a marine plant. The land plant refers to a plant that grows naturally on land, and the marine plant refers to a plant that grows in the water of the sea or in the soil of the sea.

In the present invention, the land plant or marine plant can be divided into above-ground plants, underground plants, and outpost plants according to the intended use area, but the use of plants of the same type is different depending on the purpose of use, so it must be clearly distinguished. no.

In the present invention, the above-ground plant refers to a plant using a stem, leaf, flower, fruit, fruit, or seed blood (bak) of the plant. However, it is not limited only to the stem, leaf, flower, fruit, fruit, or seed blood (garden) of the plant.

In addition, the underground plant refers to a plant using a root of a plant or a stem growing underground.

In the present invention, the outpost plant refers to a plant using both the above-ground and underground portions of the plant.

In one embodiment of the present invention, the finely pulverized plant powder is a finely pulverized land plant powder.

In another embodiment of the present invention, the oxidized pulverized plant powder of the composition may be included in a concentration of 0.1-30% (w/v) based on the total weight of the plant gel cosmetic composition.

As a result of preparing the plant gel for each land plant/marine plant for use, the present inventors found that the concentration of the oxidized finely pulverized plant powder in which the gel is well formed is different for each region of each land plant. For example, in the case of outposts, roots, stems, or their blood (leaf) of terrestrial plants, the gel should contain 1-25% (w/v) of oxidized pulverized plant powder based on the total weight of the composition. It is formed, and in the case of leaves, fruits, seeds, or their blood (garden) of land plants, a concentration higher than that of the outpost, root, and stem based on the total weight of the composition is 5-30% (w/v) The gel is well formed only when the oxidized finely pulverized plant powder is included, and in the case of marine plants, the gel is well formed when the oxidized finely pulverized plant powder is contained at a lower concentration of 1-20% (w/v).

Accordingly, in one embodiment of the present invention, the land plant is an outpost, root, stem, or blood (bak) of a land plant, wherein the composition is 1-25% (w/) based on the total weight of the composition. v) oxidized pulverized plant powder.

When the composition contains the oxidized pulverized plant powder of the outpost, root, stem, or their blood (leaf) of land plants, 1-25, 1-20, based on the total weight of the composition, 1-15, 1-10, 3-25, 3-20, 3-15, 3-10, 5-25, 5-20, 5-15, 5-10, 10-25, 10-20, 10- 15, 15-25, 15-20% (w/v) of oxidized finely pulverized plant powder may be included, but the present invention is not limited thereto. However, when the composition of the present invention contains the oxidized pulverized plant powder of land plant outposts, roots, stems, or their blood (leaf), plants at a concentration of at least 1% (w/v) or more It was confirmed that the gel formation was good only when the powder was included. In addition, when the plant powder exceeds 25% (w/v), the concentration is too high and the feeling of use is not good.

In one embodiment of the present invention, the outposts, roots, or blood (bak) of the land plants are red ginseng, ginseng, honggyeongcheon, angelica, hwanggi, baekchul, wooseul, dangsam, sanyangsam, deodeok, swandae, bellflower, sapju , Arrowroot, Baek Hasuo, Hemp, Donggulle, Tobokryeong, Cheongung, Licorice, Peony, Rehmannia, and their blood (bak), but are not limited thereto.

In another embodiment of the present invention, the land plant is motherwort, Samjiguyeopcho, Gujeolcho, Lamiaceae, celandine, etc. are outpost plants, mainly leaves and stems of the plant are used, but all the leaves and stems, roots and flowers, and even fruits It can be used including.

In one embodiment of the present invention, the stem of the land plant or the blood (bak) thereof is a rapeseed, ivy, sugar cane, cinnamon tree, yellow wall tree, black oak, bamboo, centella, and their It is selected from the group consisting of blood (bak), but is not necessarily limited thereto.

In another embodiment of the present invention, Centella asiatica, eryngii flower, thistle, gorilla, etc. are above-ground plants and mainly use both stems and leaves, but only the stems as described above, or only the leaves as described later may be used.

In a specific embodiment of the present invention, the outpost, root, stem, or blood (bak) of the land plant is selected from the group consisting of red ginseng, ginseng, centella, and blood (bak) thereof, and the composition is It can be used for skin moisturizing purposes.

As demonstrated in the examples of the present invention, the red persimmon gel, ginseng gel, and centella perilla gel of the present invention can be usefully used as a composition for skin moisturizing because they increase the expression of factors related to moisturizing in the skin epidermis.

In a specific embodiment of the present invention, the outpost, root, stem, or blood (bak) of the land plant is selected from the group consisting of ginseng, red ginseng, centella, and blood (bak) thereof, and the composition is It can be used to strengthen skin barrier.

As demonstrated in the examples of the present invention, the red ginseng gel and centellar gel of the present invention increase the expression of factors related to skin barrier reinforcement, and thus can be usefully used as a composition for skin barrier reinforcement.

In a specific embodiment of the present invention, the outpost, root, stem, or blood (bak) of the land plant is selected from the group consisting of red ginseng, ginseng, rapeseed, centella asiatica, cheopseokgok, and their blood (bak) And, the composition can be used for preventing skin aging.

As demonstrated in the examples of the present invention, the red ginseng gel, ginseng gel, rapeseed gel, centella gel, and cheopseokgok gel of the present invention reduce the expression level of genes related to skin wrinkles, and the expression level of genes related to skin elasticity. As it increases, it can be usefully used as a composition for preventing skin aging.

In a specific embodiment of the present invention, the outposts, roots, stems, or their blood (bak) of the land plant is selected from the group consisting of ginseng, centella, and their blood (bak), and the composition is UV-protected Can be used for purposes.

As demonstrated in the examples of the present invention, the ginseng gel and centripetal gel of the present invention are excellent in reducing the expression level of genes related to skin wrinkles after irradiation with ultraviolet rays (UVA and UVB), improving cell function, and regenerating cells, It can be usefully used as a composition for UV protection.

In a specific embodiment of the present invention, the outposts, roots, stems, or their blood (leaf) of the land plant are cheolpiseokgok, and the composition may be used for skin whitening.

As demonstrated in the examples of the present invention, the cheopseokgok gel of the present invention decreases the expression of genes related to melanogenesis, and thus can be usefully used as a composition for skin whitening.

In another embodiment of the present invention, the land plant is a leaf, a fruit, a seed, or the blood of a land plant, wherein the composition is 5-30% (w/v) based on the total weight of the composition Oxidized finely ground plant powder.

When the composition contains oxidized pulverized plant powder of leaves, fruits, seeds of land plants, or their blood (baked), 5-30, 5-25, based on the total weight of the composition, 5-20, 5-15, 5-10, 10-30, 10-25, 10-20, 10-15, 15-30, 15-25, 15-20, 20-30, 25-30%(w /v) of the oxidized finely ground plant powder may be included, but is not limited thereto. However, when the composition of the present invention contains the oxidized pulverized plant powder of the leaves, fruits, seeds of land plants, or their blood (baked), plants at a concentration of at least 5% (w/v) or more It was confirmed that the gel formation was good only when the powder was included. In addition, when the plant powder exceeds 30% (w/v), the concentration is too high and the feeling of use is not good.

In one embodiment of the present invention, the leaves of the land plant are selected from the group consisting of camellia leaves, green tea leaves, mulberry leaves, and mint leaves, but are not limited thereto.

In one embodiment of the present invention, the fruits, seeds, or blood (bak) of the land plants are Jeju blue beans, Jangdan beans, kidney beans, small beans, lentils, peas, seoritae, seomoktae, eastern beans, peanuts, mung beans, Beans of legumes such as chickpea and red beans, pomegranate, schisandra chinensis, goji berry, cornus, scallionate, berry, jujube, apricot, tangerine, loquat fruit, oleander fruit, oleum, stone pear, quince, bokbunja, raspberries and their blood ( Park) is selected from the group consisting of, but is not necessarily limited thereto. The fruit includes not only the above-described blood, but also the flesh.

In a specific embodiment of the present invention, the leaves, fruits, seeds, or blood (bak) of the land plant are camellia leaves, green tea leaves, or a combination thereof, and the composition may be used for skin moisturizing. .

As demonstrated in the examples of the present invention, the camellia leaf gel and the green tea leaf gel of the present invention can be usefully used as a composition for skin moisturizing because they increase the expression of factors related to moisturizing in the skin epidermis.

In a specific embodiment of the present invention, the leaves, fruits, seeds, or blood (bak) thereof of the land plant is selected from the group consisting of camellia leaves, green tea leaves, Jeju green beans, and blood (bak) thereof, The composition may be used for preventing skin aging.

As demonstrated in the examples of the present invention, the camellia leaf gel, green tea leaf gel, and Jeju blue bean gel of the present invention reduce the expression level of genes related to skin wrinkles and increase the expression level of genes related to skin elasticity, thus preventing skin aging. It can be usefully used as a composition.

In a specific embodiment of the present invention, the leaves, fruits, seeds, or blood (bak) of the land plant are Jeju blue beans, and the composition may be used for skin whitening.

As demonstrated in the examples of the present invention, since the Jeju blue bean gel of the present invention reduces the expression of genes related to melanogenesis, it can be usefully used as a composition for skin whitening.

In another embodiment of the present invention, the finely pulverized plant powder is a finely pulverized marine plant powder.

In one embodiment of the present invention, the marine plant will contain 1-20% (w / v) of oxidized finely pulverized plant powder based on the total weight of the composition, plant gel cosmetic composition. When the composition contains oxidized finely pulverized plant powder of marine plants, 1-20, 1-15, 1-10, 3-20, 3-15, 3-10, 5 based on the total weight of the composition -20, 5-15, 5-10, 10-20, 10-15, 15-20% (w/v) oxidized finely pulverized plant powder may be included, but is not limited thereto. However, when the composition of the present invention contains the oxidized finely pulverized plant powder of marine plants, it was confirmed that gel formation was well performed when the plant powder was included in a concentration of at least 1% (w/v) or more. In addition, when the plant powder exceeds 20% (w/v), the concentration is too high and the feeling of use is not good.

In one embodiment of the present invention, the marine plant means a marine benthic plant. Marine benthic plants are classified into seaweed (macroalgae) and marine angiosperms.

In the present invention, the seaweed (macroalgae) is classified into green algae, brown algae, red algae, blue algae, and the like. Green algae include thorny green, single green, plum saengyi, and hearing, brown algae include seaweed, kelp, hathaban, haricot, Ecklonia cava, and earthworm. However, it is not limited thereto.

In the present invention, the marine flowering plants are classified into sea grasses, salt marsh plants, and mangroves. Seaweeds include turtle horses, algae, and horsetails, and salt marsh plants include dyed grasses, rushes, and dicotyledonous shrubs, but are not limited thereto.

In a specific embodiment of the present invention, the marine plant is an agar, and the composition may be used for skin moisturizing.

As demonstrated in the examples of the present invention, the agar gel of the present invention can be usefully used as a composition for skin moisturizing because it increases the expression of factors related to moisturizing in the skin epidermis.

In a specific embodiment of the present invention, the marine plant is an agar, and the composition may be used for preventing skin aging.

As demonstrated in the examples of the present invention, the agar gel of the present invention can be usefully used as a composition for preventing skin aging because it reduces the expression level of genes related to skin wrinkles and increases the expression level of genes related to skin elasticity. .

In a specific embodiment of the present invention, the marine plant is Ecklonia cava, and the composition may be used for skin whitening.

As demonstrated in the examples of the present invention, the Ecklonia gel of the present invention reduces the expression of genes related to melanogenesis, and thus can be usefully used as a composition for skin whitening.

The plant gel composition according to the present invention is prepared by a manufacturing method comprising the following steps:

(a) reacting by adding at least one oxidizing agent to the finely ground plant powder;

(b) purifying the plant powder and diluting it in purified water;

(c) Homogenizing the diluted solution of plant powder and concentrating or filtering to prepare a plant gel.

In the plant gel production step of the present invention, the particle diameter of the finely pulverized plant powder in step (a) is 1 to 300 μm or less.

In one embodiment of the present invention, the oxidizing agent is hypohalogenic acid or a metal salt thereof, or ahalogenic acid or a metal salt thereof, and the "halogen" of the hypohalogenic acid or a metal salt thereof, and ahalogenic acid or a metal salt thereof is fluorine, These are chlorine, bromine and iodine.

In another embodiment of the present invention, the metal of the hypohalogenate metal salt and the hypohalogenate metal salt is an alkali metal such as lithium, potassium, and sodium; Alkaline earth metals such as calcium, magnesium, and strontium, and more specifically, the hypohalogenate is sodium hypochlorite, potassium hypochlorite, calcium hypochlorite, magnesium hypochlorite, strontium hypochlorite, and ammonium hypochlorite, but is not limited thereto.

In the plant gel preparation step of the present invention, the step (a) is performed by adding a nitroxy radical compound as a catalyst.

In one embodiment of the present invention, the nitroxy radical compound is (2,2,6,6-Tetramethylpiperidin-1-yl)oxyl or a derivative thereof, and the derivative of the nitroxy radical compound is specifically 4-Acetamido- 2,2,6,6-tetramethylpiperidine 1-oxyl; 4-Amino - 2,2,6,6-tetramethylpiperidine- d 17 -1-oxyl; 4-Amino-2,2,6,6-tetramethylpiperidine-1-oxyl; 2-Azaadamantane-N-oxyl; 4-Carboxy-2,2,6,6-tetramethylpiperidine 1-oxyl; 4-Hydroxy-2,2,6,6-tetramethylpiperidine 1-oxyl; 4-Hydroxy-2,2,6,6-tetramethylpiperidine-d ₁₇ -1-oxyl; 4-Hydroxy-2,2,6,6-tetramethylpiperidine 1-oxyl benzoate; 4-(2-Iodoacetamido)-2,2,6,6-tetramethyl-1-piperidinyloxy; 4-Isothiocyanato-2,2,6,6-tetramethylpiperidine 1-oxyl; 4-Maleimido-2,2,6,6-tetramethyl-1-piperidinyloxy; 4-Methoxy-2,2,6,6-tetramethyl-1-piperidinyloxy; 4-Oxo-2,2,6,6-tetramethyl-1-piperidinyloxy; 4-Oxo-, 2,2,6,6-tetramethylpiperidine- d 16 -1-oxyl; 4-Phosphonooxy-2,2,6,6-tetramethyl-1-piperidinyloxy; 4-Methacryloyloxy-2,2,6,6-tetramethylpiperidine-1-oxyl; And 2,2,6,6-Tetramethyl-4-(methylsulfonyloxy)-1-piperidinooxy may be one or more compounds selected from the group consisting of, but is not limited thereto.

In the plant gel preparation step of the present invention, the purification of step (b) includes adding alcohol to the reaction solution of step (a) to precipitate the plant powder, and then filtering the precipitated plant powder. The alcohol is a C1 to C5 low-cost alcohol.

In the plant gel production step of the present invention, the purification of step (b) includes the step of membrane filtration by adding purified water to the reaction solution of step (a).

In the plant gel production step of the present invention, the homogenization in step (c) is performed at a pressure of 800 to 1000 bar.

In addition, in the plant gel production step of the present invention, the concentration in step (c) is carried out by concentration under reduced pressure at 50-65°C.

In the plant gel preparation step of the present invention, the filtration of step (c) is performed by membrane filtration.

In the plant gel preparation step of the present invention, the plant powder content of the plant gel prepared in step (c) is 0.1-30% (w/v).

Hereinafter, the method for preparing the plant gel of the present invention will be described in detail.

Step (a): Oxidation step of finely ground plant powder

First, a finely ground plant powder is obtained. The finely ground plant powder can be purchased or obtained by grinding the dried plant with a grinder.

In the present invention, the finely pulverized plant powder in step (a) may have a particle diameter of 1 to 300 μm or less. The smaller the particle size of the finely pulverized plant powder has the advantage that the oxidation reaction is performed at a high rate in a short time.

In the step of the present invention, an oxidation reaction is performed by adding at least one oxidizing agent to the plant powder. The oxidizing agent may be hypohalogenic acid or a metal salt thereof, or ahalogenic acid or a metal salt thereof.

In one embodiment of the present invention, the hypohalogenic acid or its metal salt, and the halogen in the subhalogenic acid or its metal salt may be chlorine, bromine, or iodine. Therefore, it may be specifically hypochlorous acid, hypobromic acid, hypoiodic acid, chlorous acid, abromic acid, and aiodic acid.

In another embodiment of the present invention, the metal in the hypohalogenate metal salt and the hypohalogenate metal salt is an alkali metal such as lithium, potassium, and sodium; It may be an alkaline earth metal such as calcium, magnesium, and strontium. Therefore, specific examples of the hypohalogenate include sodium hypochlorite, potassium hypochlorite, calcium hypochlorite, magnesium hypochlorite, strontium hypochlorite, ammonium hypochlorite, and the like. Specific examples of the hypochlorite include lithium chlorite, potassium chlorite, sodium chlorite, calcium chlorite, magnesium chlorite, strontium chlorite, and ammonium chlorite. In addition, a bromate salt or an aiodate salt corresponding to the chlorite salt may be used.

The oxidation reaction in this step can be carried out by adding a nitroxy radical compound as a catalyst. The "nitroxy radical compound" may be a nitrooxy radical compound or a derivative thereof. Specifically, the nitroxy radical compound is (2,2,6,6-Tetramethylpiperidin-1-yl)oxyl, and the derivative of the nitroxy radical compound is 4-Acetamido-2,2,6,6-tetramethylpiperidine 1-oxyl; 4-Amino-2,2,6,6-tetramethylpiperidine- d 17 -1-oxyl; 4-Amino-2,2,6,6-tetramethylpiperidine-1-oxyl; 2-Azaadamantane-N-oxyl; 4-Carboxy-2,2,6,6-tetramethylpiperidine 1-oxyl; 4-Hydroxy-2,2,6,6-tetramethylpiperidine 1-oxyl; 4-Hydroxy-2,2,6,6-tetramethylpiperidine-d ₁₇ -1-oxyl; 4-Hydroxy-2,2,6,6-tetramethylpiperidine 1-oxyl benzoate; 4-(2-Iodoacetamido)-2,2,6,6-tetramethyl-1-piperidinyloxy; 4-Isothiocyanato-2,2,6,6-tetramethylpiperidine 1-oxyl; 4-Maleimido-2,2,6,6-tetramethyl-1-piperidinyloxy; 4-Methoxy-2,2,6,6-tetramethyl-1-piperidinyloxy; 4-Oxo-2,2,6,6-tetramethyl-1-piperidinyloxy; 4-Oxo-, 2,2,6,6-tetramethylpiperidine- d 16 -1-oxyl; 4-Phosphonooxy-2,2,6,6-tetramethyl-1-piperidinyloxy; 4-Methacryloyloxy-2,2,6,6-tetramethylpiperidine-1-oxyl; And 2,2,6,6-Tetramethyl-4-(methylsulfonyloxy)-1-piperidinooxy may be one or more compounds selected from the group consisting of.

The oxidation reaction in this step may be carried out by adding a salt of a halogen and an alkali metal (alkali metal salt), a salt of a halogen and an alkaline earth metal (alkaline earth metal salt), and an ammonium salt as a cocatalyst.

Examples of the halogen for forming the alkali metal salt, alkaline earth metal salt and ammonium salt include chlorine, bromine, and iodine. Examples of the alkali metal forming the alkali metal salt include lithium, potassium, and sodium. Examples of the alkaline earth metal forming the alkaline earth metal salt include calcium, magnesium, and strontium.

More specifically, lithium bromide, potassium bromide, sodium bromide, lithium iodide, potassium iodide, sodium iodide, lithium chloride, potassium chloride, sodium chloride, calcium bromide, magnesium bromide, strontium bromide, calcium iodide, magnesium iodide, calcium iodide, strontium Magnesium chloride, strontium chloride, and the like, but are not limited thereto. Moreover, ammonium bromide, ammonium iodide, and ammonium chloride are mentioned as an ammonium salt. These cocatalysts can be used alone or in combination of two or more, and these cocatalysts may form a hydrate.

The oxidation reaction of step (a) of the present invention may be carried out for 2 to 4 hours while maintaining the pH at 9-11.

Step (b): Purification and dilution of the oxidized plant powder

Subsequently, in order to remove unreacted raw materials and impurities (raw materials such as nitroxy radicals used in the reaction, NaBr, NaClO, and reaction adducts such as NaCl) from the oxidized plant powder, the plant powder is purified and further microparticles are made (homogenized). For this, it is diluted in purified water.

According to one embodiment of the present invention, the purification of step (b) may be performed by adding alcohol to the reaction solution of step (a) to precipitate plant powder, and then filtering the precipitated plant powder. Here, the alcohol may be a low-cost alcohol of C1 to C5, and the addition of alcohol-precipitation of plant powder-filtration step may be performed by repeating two or more times.

According to another embodiment of the present invention, the purification of step (b) may be performed by adding purified water to the reaction solution of step (a) and filtering the membrane (ultrafiltration). If the membrane filtration method is used, unreacted raw materials and impurities generated in the oxidation step of plant powder can be removed in a single process without performing the alcohol precipitation and purified water dilution steps.

Step (c): Homogenization of plant powder diluent and preparation of plant gel

Finally, the plant powder diluted solution was homogenized (microparticleized) through a high-pressure homogenizer (microfluidizer) and concentrated or filtered to prepare the plant gel of the present invention. The homogenization in step (c) may be performed at a pressure of 800 to 1000 bar.

According to an embodiment of the present invention, the concentration after homogenization may be performed by concentration under reduced pressure or vacuum concentration at 50-65°C.

According to another embodiment of the present invention, the filtering step after homogenization may be performed by membrane filtration in the same manner as in step (b).

In addition, the plant powder content of the finally prepared plant gel may be 0.1-30% (w/v). According to an embodiment of the present invention, when the microparticleized oxidized plant powder is adjusted to have a concentration of 0.1-30% (w/v), it may exhibit a gel state and exhibit the functionality of the plant raw material itself. When the plant powder content of the prepared plant gel is less than 0.1% (w/v), it is difficult to expect the efficacy of the plant raw material itself according to the addition of the plant powder, and when it exceeds 30% (w/v), the effect is increased compared to the amount added. It is not economical because it is insignificant.

According to another aspect of the present invention, the present invention provides a plant gel comprising 0.1-30% (w/v) of oxidized pulverized plant powder and a residual amount of a solvent.

When the plant gel of the present invention is prepared as a cosmetic composition, the cosmetic composition of the present invention is a solution, suspension, emulsion, paste, lotion, gel, water-soluble liquid, cream, essence, surfactant-containing cleansing, oil, spray, water. Heavy oil (O/W) type or water-in-oil (W/O) type of basic cosmetic formulation; Color cosmetics formulations such as oil-in-water or water-in-oil makeup base, foundation, skin cover, powder, lipstick, lip gloss, eye shade, cheek color or eyebrow pencil; And, it can be prepared in any one formulation selected from the formulation for the scalp. In addition, the cosmetic composition of the present invention may optionally be applied to the skin in the form of an aerosol.

When the formulation of the cosmetic composition of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide, etc. are used as carrier components. Can be used.

When the formulation of the present invention is a powder or 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, a solubilizing agent or an emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan can be used.

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

When the formulation of the present invention is a surfactant containing cleansing, as a carrier component, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters may be used.

Components included in the cosmetic composition of the present invention include ingredients commonly used in cosmetic compositions, in addition to plant gels and carrier components, and include conventional auxiliary agents such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances. Can include. The amount of the plant gel in the cosmetic composition of the present invention is not particularly limited, and is preferably included in an amount sufficient to achieve the efficacy of the plant raw material.

The present invention relates to the use of a gel comprising plants as raw materials. The plant gel of the present invention can produce a gel-like composition using plant powder as a raw material through the same process collectively for various plant types without the need for individual extraction and fractionation processes according to plant types like conventional plant extract compositions. As a cosmetic composition, it has excellent process efficiency in that it has excellent use as a cosmetic composition such as skin moisturizing effect, skin barrier strengthening effect, skin aging prevention effect, UV protection effect, and whitening effect depending on the raw material components of each plant gel. It can be useful.

Figures 1a and 1b is a culture of human normal fibroblasts by adding centella asiatica gel at a concentration of 0, 5, 10, 20 μg/ml in order to investigate the cell regeneration efficacy of centella asiatica gel prepared from centella asiatica among the plant gels of the present invention. It is a diagram showing the results.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for describing the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

Example

Throughout the present specification, "%" used to indicate the concentration of a specific substance is (weight/weight)% for solids/solids, (weight/volume)% for solids/liquids, and Liquid/liquid is (vol/vol) %.

Experimental material

Materials used for the plant gel composition of the present invention are as follows. Red ginseng, ginseng, rapeseed, centella asiatica, camellia leaves, green tea leaves, Jeju green beans, cheopseokgok, Ecklonia cava, and agar are It was purchased and used in Samdo P&F Co., Ltd. (Korea) and herbal medicine market. Alfa Aesar's (2,2,6,6-Tetramethylpiperidin-1-yl)oxyl (TEMPO) reagent was used, and in the case of NaClO, Daejunghwa Geum Co., Ltd., NaBr, NaOH, HCl, and ethanol were manufactured by Duksan Pharmaceutical The product was used.

Example 1: Preparation of the red ginseng gel composition of the present invention

The dried red ginseng was pulverized with a grinder and then sieved with 150 mesh to obtain red ginseng powder of 100 μm or less. 300 mL of purified water was added to 15 g of the pulverized red ginseng powder and stirred. After that, 0.47 g of TEMPO and 3.09 g of NaBr were added and dissolved. After adding 100 mL of NaClO dropwise, it was oxidized at room temperature for 15 hours while maintaining the pH to 10-11 with NaOH. The red ginseng powder was purified by adding HCl to the oxidized red ginseng reaction solution to neutralize the pH to 6-7, and removing the raw material and the reaction part product used in the oxidation reaction. Purification was largely performed in two ways.

In the first method (ethanol precipitation method), 300 mL of ethanol was added to the reaction solution to precipitate, followed by filtration to obtain a filtrate. The filtrate was added to 50% ethanol, stirred, washed, and then filtered and purified. As the second method (membrane filtration method), the reaction solution was purified by continuously adding purified water using a membrane filter (FMX-B5, a kind of UF, Bugang Tech Co., Ltd.) until impurities were removed.

3 L of purified water was added to the purified red ginseng powder and dispersed by stirring at 2500 RPM for 30 minutes using a homogenizer (PRIMIX, ROBOMIXER). The red ginseng powder dispersion was micronized by passing it 5 times at 800 to 1000 bar using a high pressure homogenizer (M-110Y microfluidizer, Microfluidics Corporation, USA). The refined red ginseng composition was concentrated with a vacuum concentrator (R-210 set, BUCHI) or a membrane filter (FMX-B5, Bugang Tech Co., Ltd.) to obtain 205 g of the red ginseng gel of the present invention.

Example 2: Preparation of ginseng gel

The dried ginseng was pulverized with a grinder and then sieved with 150 mesh to obtain a ginseng powder of 100 μm or less. 15 g of the pulverized ginseng powder was prepared in the same manner as in Example 1 to obtain 210 g of the ginseng gel of the present invention.

Example 3: Preparation of rapeseed gel

The dried rapeseed was pulverized with a grinder and then sieved with 150 mesh to obtain rapeseed powder of 100 μm or less. 15 g of the pulverized rapeseed powder was prepared in the same manner as in Example 1 to obtain 125 g of the rapeseed gel of the present invention.

Example 4: Preparation of Centella asiatica gel

After pulverizing the dried centella asiatica with a grinder, it was sieved with 150 mesh to obtain centella asiatica powder of 100 μm or less. 15 g of the pulverized Centella asiatica powder was prepared in the same manner as in Example 1 to obtain 160 g of Centella asiatica gel of the present invention.

Example 5: Preparation of camellia leaf gel

The dried camellia leaves were pulverized with a grinder and then sieved with 150 mesh to obtain camellia leaf powder of 100 μm or less. 15 g of the pulverized camellia leaf powder was prepared in the same manner as in Example 1 to obtain 108 g of the camellia leaf gel of the present invention.

Example 6: Preparation of green tea leaf gel

The dried green tea leaves were pulverized with a grinder and then sieved with 150 mesh to obtain green tea leaf powder of less than 100 μm. 15 g of the pulverized green tea leaf powder was prepared in the same manner as in Example 1 to obtain 55 g of green tea leaf gel of the present invention.

Example 7: Preparation of Jeju Blue Bean Gel

The dried Jeju green beans were pulverized with a grinder and then sieved with 150 mesh to obtain a Jeju blue bean powder of 100 μm or less. 15 g of the pulverized Jeju blue bean powder was prepared in the same manner as in Example 1 to obtain 40 g of the Jeju blue bean gel of the present invention.

Example 8: Preparation of iron crumb gel

After pulverizing the dried iron crumbs with a grinder, they were sieved with 150 mesh to obtain iron crumbs of less than 100 μm. 15 g of the pulverized iron pyrite powder was prepared in the same manner as in Example 1 to obtain 285 g of the iron pyrite grain gel of the present invention.

Example 9: Preparation of persimmon taegel

After the dried Ecklonia cava was pulverized with a grinder, it was sieved with 150 Mesh to obtain Ecklonia cava powder of 100 μm or less. 15 g of the pulverized Ecklonia cava powder was prepared in the same manner as in Example 1 to obtain 190 g of Ecklonia cava gel of the present invention.

Example 10: Preparation of agaric gel

After pulverizing the dried agar-agar with a grinder, 15 g of powder was prepared in the same manner as in Example 1 to obtain 204 g of agar-agar gel of the present invention.

Example 11: Preparation of pomegranate gel

After pulverizing the dried pomegranate with a grinder, 15 g of powder was prepared in the same manner as in Example 1 to obtain 185 g of the pomegranate gel of the present invention.

Example 12: Preparation of honggyeongcheon gel

After pulverizing the dried rhododendron cloth with a grinder, 15 g of powder was prepared in the same manner as in Example 1 to obtain 110 g of the rhododendron gel of the present invention.

Example 13. Method 1 for purifying plant powder of the present invention (ethanol precipitation method)

After the TEMPO reaction, the same amount of ethanol relative to the volume of the reaction solution was added, stirred, and then filtered. The filtered wet cake was added with 50% ethanol in the same volume (reaction solution + ethanol) of the previous step, stirred, and then filtered. After the filtered wet cake was diluted with purified water, homogenization was performed 5 times with a high pressure homogenizer (Microfluidizer). The microparticleized plant powder solution was concentrated under reduced pressure to obtain a plant gel containing 3-30% solids.

Example 14. Method 2 for purifying plant powder of the present invention (membrane filtration method)

After the TEMPO reaction, the reaction solution is removed using a membrane filter (FMX-B5, Bugang Tech Co., Ltd.) to remove unreacted raw materials and impurities (raw materials such as TEMPO, NaBr, NaClO used in the reaction and reaction adducts NaCl, etc.), and then purified water. Diluted using. The diluted solution was homogenized (micronized) 5 times using a high pressure homogenizer (Microfluidizer). The finely granulated plant powder solution was again removed by water using a membrane filter (FMX-B5, Bugang Tech Co., Ltd.) to obtain a plant gel containing 3-30% solids.

Experimental example

Experimental Example 1: Evaluation of skin moisturizing efficacy of plant gel

Experimental example 1-1: water channel that supplies moisture to the skin surface Aqua On porin-3 (APQ-3) Korean gene expression level investigation

For the evaluation of skin moisturizing efficacy, an experiment was performed to confirm the expression of aquaporin-3 (AQP-3) gene. Normal human keratinocytes (gibco) were inoculated with 4.5X10 ⁵ cells in a 60 mm culture dish (corning), cultured for 1 day (37° C., CO ₂ 5%), and then replaced with Serum-free medium (Epilife, gibco). After 24 hours, the samples were treated by concentration and incubated in an incubator at 37° C. and CO ₂ 5% for 24 hours. After culture, the cells were collected with TRIzol and RNA was isolated according to the manufacturer's method. After quantifying the isolated RNA, cDNA was synthesized with 1 μg of RNA, followed by real-time PCR. Aquaporin-3 and β-Actin primers used in PCR were synthesized and used by Cosmo Genetec (Korea), and the primer sequence is shown in Table 1. Retinoic acid was used as a positive control. Table 2 shows the results of the expression level.

Primers Primer Sequence (5' to 3') SEQ ID NO β-actinsense GGC ACC CAG CAC AAT GAA G One β-actin
antisense CCG ATC CAC ACG GAG TAC TTG 2 Aquaporin-3(AQP-3)Sense GGG ACC AGT CGG AAG GGA T 3 Aquaporin-3(AQP-3)Antisense CAC AGA TGG ACA GGC TGC CT 4

AQP-3 mRNA expression level Test group Experimental concentration (μg/ml) AQP-3 mRNA
Expression rate (%) Control - 100.0 Retinoic acid 100 nM 135.5 Example 1
(Red Ginseng Gel) 10 111.4 25 115.1 50 145.4 Example 5
(Camellia Leaf Gel) 10 161.36 25 197.06 Example 6
(Green Tea Leaf Gel) 25 492.37 50 504.65 100 430.31 Example 10
(Agarigel) 5 197.83 10 281.45 25 204.86

As a result of confirming the expression of the moisturizing-related AQP-3 gene of the plant gel, it was confirmed that the expression of the gene was increased at various concentrations in Examples 1, 5, 6, and 10, thereby having a skin moisturizing effect.

Experimental example 1-2: Skin moisturizing Hyaluronic acid mechanism-related gene Expression amount investigation

1) hyaluronic acid synthase Hyaluronic acid synthase -2 (HAS- 2) on Korean gene expression level investigation

In order to evaluate skin moisturizing efficacy, an experiment was performed to confirm the gene expression of Hyaluronic acid synthase-2 (HAS-2). Normal human keratinocytes (gibco) were inoculated with 4.5X10 ⁵ cells in a 60 mm culture dish (corning), cultured for 1 day (37° C., CO ₂ 5%), and replaced with Serum-free medium (Epilife, gibco). After 24 hours, the samples were treated by concentration and incubated in an incubator at 37° C. and 5% CO ₂ for 24 hours. After culture, the cells were collected with TRIzol and RNA was isolated according to the manufacturer's method. After quantifying the isolated RNA, cDNA was synthesized with 1 μg of RNA, followed by real-time PCR. Hyaluronic acid synthase-2 (HAS-2) and β-Actin primers used in PCR were synthesized and used by Cosmo Genetec (Korea), and the primer sequences are shown in Table 3. Retinoic acid was used as a positive control. The results of the expression level are shown in Table 4.

HAS-2 Primers Primer Sequence (5' to 3') SEQ ID NO Hyaluronic acid-2 (HAS-2) Sense ACA ACA CCT TAG TTC CTC TA 5 Hyaluronic acid-2 (HAS-2) antisense AGC AGT GAT ATG TCT CCT 6 β-actin Sense GGC ACC CAG CAC AAT GAA G One β-actin antisense CCG ATC CAC ACG GAG TAC TTG 2

HAS-2 mRNA expression level Test group Experimental concentration (μg/ml) HAS-2 mRNA
Expression rate (%) Control - 100.0 Retinoic acid 100 nM 165.1 Example 1
(Red Ginseng Gel) 10 111.3 25 120.0 50 127.5

As a result of confirming the expression of the HAS-2 gene related to the moisturizing of the plant gel, it was confirmed that the treatment of Example 1 had a skin moisturizing effect as the gene expression was increased in a concentration dependent manner.

2) Investigation of gene expression level for hyaluronic acid receptor CD-44

An experiment was conducted to confirm the gene expression of CD-44, a receptor for hyaluronic acid, which is one of the moisturizing factors contributing to skin hydration. Normal human keratinocytes (gibco) were inoculated with 4.5X10 ⁵ cells in a 60 mm culture dish (corning), cultured for 1 day (37° C., CO ₂ 5%), and replaced with Serum-free medium (Epilife, gibco). After 24 hours, the samples were treated by concentration and incubated in a 37°C, CO ₂ 5% incubator for 24 hours. After culture, the cells were collected with TRIzol and RNA was isolated according to the manufacturer's method. After quantifying the isolated RNA, cDNA was synthesized with 1 μg of RNA, followed by real-time PCR. For PCR, CD-44 and β-Actin primers were synthesized and used by Cosmo Genetec (Korea), and the primer sequence is shown in Table 5. Retinoic acid was used as a positive control. The results of the expression level are shown in Table 6.

CD-44 Primers Primer Sequense (5' to 3') SEQ ID NO CD-44 sense GCT ATT GAA AGC CTT GCA GAG 7 CD-44 antisense CGC AGA TCG ATT TGA ATA TAA CC 8 β-actin sense GGC ACC CAG CAC AAT GAA G One β-actin antisense CCG ATC CAC ACG GAG TAC TTG 2

CD-44 mRNA expression level Test group Experimental concentration (μg/ml) CD-44 mRNA
Expression rate (%) Control - 100.0 Retinoic acid 50 nM 134.13 Example 6
(Green Tea Leaf Gel) 25 145.10 50 161.70

As a result of confirming the expression of the moisturizing-related CD-44 gene of the plant gel, it was confirmed that the treatment of Example 6 had a skin moisturizing effect as the gene expression was increased in a concentration-dependent manner.

Experimental example 1-3: Evaluation of moisturizing efficacy for the synthesis of natural moisturizing factor (NMF) in the skin epidermis

1) Natural moisturizing factor in the skin epidermis Transcript Profila green (Pro Filaggrin , Pro FLG) expression

For the evaluation of skin moisturizing efficacy, an experiment was conducted to confirm the gene expression of the natural moisturizing factor prewashed car Pro filaggrin (Pro FLG). Normal human keratinocytes (gibco) were inoculated with 4.5X10 ⁵ cells in a 60 mm culture dish (corning), cultured for 1 day (37° C., CO ₂ 5%), and replaced with Serum-free medium (Epilife, gibco). After 24 hours, the samples were treated by concentration and incubated in a 37°C, CO ₂ 5% incubator for 24 hours. After culture, the cells were collected with TRIzol and RNA was isolated according to the manufacturer's method. After quantifying the isolated RNA, cDNA was synthesized with 1 μg of RNA, followed by real-time PCR. Pro-filaggrin (Pro FLG) and β-Actin primers used in PCR were synthesized and used by Cosmo Genetec (Korea), and the primer sequences are shown in Table 7. The results of the expression level are shown in Table 8.

Pro FLG Primers Primer Sequense (5' to 3') SEQ ID NO Pro filaggrin (Pro FLG) sense CTG ATG GTA TTC AAG TTG GCT 9 Pro filaggrin (Pro FLG) antisense ACT GTG CTT TCT GTG CTT GT 10 β-actin sense GGC ACC CAG CAC AAT GAA G One β-actin antisense CCG ATC CAC ACG GAG TAC TTG 2

Pro FLG mRNA expression level Test group Experimental concentration (μg/ml) Pro FLG mRNA
Expression rate (%) Control 100.0 Example 1
(Red Ginseng Gel) 10 103.4 25 118.2 50 134.0 Example 5
(Camellia Leaf Gel) 25 143.92 Example 6
(Green Tea Leaf Gel) 25 129.50 50 154.96 100 171.06 Example 10
(Agarigel) 5 173.06 10 233.13 25 275.59

As a result of confirming the expression of the moisturizing-related Pro FLG gene of the plant gel, it was confirmed that the treatment of Example 1, Example 5, Example 6, and Example 10 had a skin moisturizing effect as gene expression increased at various concentrations.

2) Natural moisturizing factor in the skin epidermis Transcript In the expression of Filaggrin About the evaluation

Epilife (gibco) medium containing human growth factor (gibco) and 3.5×10 ⁵ human keratin cells were inoculated into 100 mm dishes and cultured at 37° C. and 5% carbon dioxide for 24 hours. After 24 hours, the samples were treated in serum-free medium for each concentration and cultured for 3 to 5 days. Calcium chloride was used as a positive control for this experiment.

After the cells were scraped off using a scraper, the pellet was settled through centrifugation. Thereafter, a protein extraction solution was added and incubated, and the protein was quantified using a QubitTM fluorometer. After adding a loading buffer to the quantified protein, it was loaded on SDS-PAGE.

After transferring using the iBlot Dry Blotting system, blocking was performed. Thereafter, primary (Filaggrin, β-actin) and secondary (GAM-HRP) antibodies were treated. After antibody treatment, west save cold ECL solution was treated, bands were photographed with chemi doc LuminoGraphII (ATTO), and the numerical values thereof were quantified and shown in Table 9.

Test group Experimental concentration (μg/ml) FLG/β-actin (%) Control - 100.0 Calcium chloride 1.2 mM 202.8 Example 1
(Red Ginseng Gel) 10 139.8 25 184.0 50 274.8 Example 2
(Ginseng gel) 50 112.1 100 184.6 Example 4
(Bung full gel) 0.5 207.9 One 262.3

When the expression of the moisturizing-related Filaggrin protein of the plant gel was confirmed, it was confirmed that the treatment of Example 1, Example 2, and Example 4 had a skin moisturizing effect as the protein expression increased at various concentrations.

3) Natural moisturizing in the epidermis Factor synthesis Enzymes involved Caspase -14 Evaluation of expression

Epilife (gibco) medium containing human growth factor (gibco) and 3.5×10 ⁵ human keratin cells were inoculated into 100 mm dishes and cultured at 37° C. and 5% carbon dioxide for 24 hours. After 24 hours, the samples were treated in serum-free medium for each concentration and cultured for 3 to 5 days. Calcium chloride was used as a positive control for this experiment.

After the cells were scraped off using a scraper, the pellet was settled through centrifugation. Thereafter, a protein extraction solution was added and incubated, and the protein was quantified using a QubitTM fluorometer. After adding a loading buffer to the quantified protein, it was loaded on SDS-PAGE.

After transferring using the iBlot Dry Blotting system, blocking was performed. Thereafter, primary (caspase-14, β-actin) and secondary (GAM-HRP) antibodies were treated. After antibody treatment, west save cold ECL solution was treated, bands were photographed with chemi doc LuminoGraphII (ATTO), and the numerical values thereof were quantified and shown in Table 10.

Test group Experimental concentration (μg/ml) Casapse-14/β-actin (%) Control - 100.0 Calcium chloride 1.2 mM 140.1 Example 2
(Ginseng gel) 25 150.1 50 156.1 100 166.3

As a result of confirming the expression of the moisturizing Caspase-14 protein in the plant gel, it was confirmed that there is a skin moisturizing effect as the protein expression increases in a concentration-dependent manner during Example 2, treatment.

4) Enzymes involved in the synthesis of natural moisturizing factors in the epidermis of the skin Bleomycin Hydrolase (Bleomycin hydrolase, BLMH) expression evaluation

Epilife (gibco) medium containing human growth factor (gibco) and 3.5×10 ⁵ human keratin cells were inoculated into 100 mm dishes and cultured at 37° C. and 5% carbon dioxide for 24 hours. After 24 hours, the samples were treated in serum-free medium for each concentration and cultured for 3 to 5 days. Calcium chloride was used as a positive control for this experiment.

After the cells were scraped off using a scraper, the pellet was settled through centrifugation. Thereafter, a protein extraction solution was added and incubated, and the protein was quantified using a QubitTM fluorometer. After adding a loading buffer to the quantified protein, it was loaded on SDS-PAGE.

After transferring using the iBlot Dry Blotting system, blocking was performed. Thereafter, primary (Bleomycin hydrolase, β-actin) and secondary (GAM-HRP) antibodies were treated. After antibody treatment, west save cold ECL solution was treated, bands were photographed with chemi doc LuminoGraphII (ATTO), and the numerical values thereof were quantified and shown in Table 11.

Test group Experimental concentration (μg/ml) BLMH/β-actin (%) Control - 100.0 Calcium chloride 1.2 mM 179.0 Example 2
(Ginseng gel) 25 117.5 50 141.7 100 169.9 Example 4
(Bung full gel) 0.5 119.8 One 119.9 5 140.6

When the expression of the moisturizing-related BLMH protein of the plant gel was confirmed, it was confirmed that the treatment of Examples 2 and 4 had skin moisturizing effects as the protein expression increased at various concentrations.

Experimental Example 2: Evaluation of the efficacy of plant gel to strengthen skin barrier

Experimental example 2-1: Plant gel Skin barrier Investigation of expression levels of tight junction proteins involved in strengthening

1) One of the tight junction proteins Occludin ( Occludin , OCDN ) Evaluation of expression

In this experimental example, in order to measure the skin barrier strengthening effect of the plant gel, a Tight Junction (TJ) protein-related gene Occludin (OCDN) expression experiment was performed that contributes decisively to the barrier function. Inoculate 4.5X10 ⁵ cells of normal human keratinocytes (gibco) into 60 mm culture dishes (corning), culture them for 1 day (37℃, CO ₂ 5%), and replace them with Serum-free medium (Epilife, gibco). After 24 hours, the samples are treated by concentration and incubated in a 37°C, CO ₂ 5% incubator for 3 to 5 days. After culture, the cells are collected with TRIzol and RNA is isolated according to the manufacturer's method. After quantifying the isolated RNA, cDNA is synthesized with 1 μg of RNA, and real-time PCR is performed. Occludin and β-Actin primers used in PCR were synthesized and used by Cosmo Genetec (Korea), and the primer sequence is shown in Table 12. Lactoeferrin was used as a positive control. Table 13 shows the results of the expression level.

OCDN Primers Primer Sequense (5' to 3') SEQ ID NO Occludin(OCDN) sense CCA CCT ATC ACT TCA GAT CAA CAA A 11 Occludin
(OCDN) antisense TTC CTG TAG GCC AGT GTC AAA AT 12 β-actin sense GGC ACC CAG CAC AAT GAA G One β-actin antisense CCG ATC CAC ACG GAG TAC TTG 2

Test group Experimental concentration (μg/ml) OCDN mRNA
Expression rate (%) Control - 100.0 Lactoferrin 0.1 μM 155.7 Example 2
(Ginseng gel) 25 114.0 50 108.5 100 118.2

As a result of confirming the expression of the skin barrier-related OCDN gene of the plant gel, it was confirmed that the expression of the skin barrier-related OCDN gene was increased in Example 2, as the gene expression was increased at various concentrations during treatment, thereby having the effect of strengthening the skin barrier.

Experimental example 2-2: Plant gel Skin moisturizing Investigation of gene expression level related to ceramide synthesis involved in barrier strengthening

1) Serine, an enzyme involved in the synthesis of ceramides Palmitoyl Transferase ( Serine Evaluation of the expression of Palmitoyl Transferase, SPT)

In this experimental example, in order to measure the skin barrier strengthening effect of the plant gel,

The expression of a gene related to the ceramide synthesis pathway that prevents water loss was confirmed. Among them, expression of serine palmitoyl transferase (SPT), an enzyme crucially involved in ceramide synthesis, was confirmed. Inoculate 4.5x10 ⁵ cells of normal human keratinocytes (gibco) in a 60 mm culture dish (corning), culture for 1 day (37°C, CO ₂ 5%), and replace with Serum-free medium (Epilife, gibco). After 24 hours, the samples are treated by concentration and incubated in a 37°C, CO ₂ 5% incubator for 3 to 5 days. After culture, the cells are collected with TRIzol and RNA is isolated according to the manufacturer's method. After quantifying the isolated RNA, cDNA is synthesized with 1 μg of RNA, and real-time PCR is performed. SPT and β-Actin primers used in PCR were synthesized and used by Cosmo Genetec (Korea), and the primer sequence is shown in Table 14. Retinoic acid was used as a positive control. The expression levels are shown in Table 15.

Primers Primer Sequense (5' to 3') SEQ ID NO Serine Palmitoyl Transferase(SPT) sense TGG TCA TTT GGC CCA GGT C 13 Serine Palmitoyl Transferase
(SPT) antisense TCC CAA CCA TTG GCT TCA CAT C 14 β-actin sense GGC ACC CAG CAC AAT GAA G One β-actin antisense CCG ATC CAC ACG GAG TAC TTG 2

Test group Experimental concentration (μg/ml) SPT mRNA
Expression rate (%) Control - 100.0 Retinoic acid 50 nM 131.7 Example 1
(Red Ginseng Gel) 10 105.3 25 111.7 50 117.1

As a result of confirming the expression of the skin barrier-related SPT gene of the plant gel, it was confirmed that the expression of the skin barrier-related SPT gene in Example 1 was increased in a concentration-dependent manner during treatment, thereby having an effect of strengthening the skin barrier.

2) Enzymes involved in ceramide synthesis Ceramide synthase -3 ( CERS3 )of Evaluation of expression

Epilife (gibco) medium containing human growth factor (gibco) and 3.5×10 ⁵ human keratin cells were inoculated into 100 mm dishes and cultured at 37° C. and 5% carbon dioxide for 24 hours. After 24 hours, the samples were treated in serum-free medium for each concentration and cultured for 3 to 5 days. Calcium chloride was used as a positive control for this experiment.

After the cells were scraped off using a scraper, the pellet was settled through centrifugation. Thereafter, a protein extraction solution was added and incubated, and the protein was quantified using a QubitTM fluorometer. After adding a loading buffer to the quantified protein, it was loaded on SDS-PAGE.

After transferring using the iBlot Dry Blotting system, blocking was performed. Thereafter, primary (CERS3, β-actin) and secondary (GAR-HRP, GAM-HRP) antibodies were treated, respectively. After antibody treatment, west save cold ECL solution was treated, bands were photographed with chemi doc LuminoGraphII (ATTO), and the numerical values thereof were quantified and shown in Table 16.

Test group Experimental concentration (μg/ml) CERS3/β-actin (%) Control - 100.0 Calcium chloride 1.8 mM 141.6 Example 1
(Red Ginseng Gel) 10 177.4 25 308.6 50 373.6 Example 4
(Bung full gel) 0.5 183.7 One 212.0 5 237.6

When the expression of the CERS3 protein related to the skin barrier of the plant gel was confirmed, it was confirmed that the treatment of Examples 1 and 4 had an effect of strengthening the skin barrier as the protein expression increased at various concentrations.

Experimental example 2-3: Plant gel Skin barrier Investigation of expression levels of tight junction proteins involved in strengthening

One) Skin barrier Enzymes involved in formation Transglutaminase -1 (Transglutaminase-1, TGase-1) gene expression evaluation

In order to measure the skin barrier strengthening effect of the plant gel, this experiment was performed to confirm the expression of transglutaminase-1 (TGase-1), one of the enzymes involved in the formation of the skin barrier. Inoculate 4.5x10 ⁵ cells of normal human keratinocytes (gibco) in a 60 mm culture dish (corning), culture for 1 day (37°C, CO ₂ 5%), and replace with Serum-free medium (Epilife, gibco). After 24 hours, the samples are treated by concentration and incubated in a 37°C, CO ₂ 5% incubator for 3 to 5 days. After culture, the cells are collected with TRIzol and RNA is isolated according to the manufacturer's method. After quantifying the isolated RNA, cDNA is synthesized with 1 μg of RNA, and real-time PCR is performed. The TGase-1 and β-Actin primers used in PCR were synthesized and used by Cosmo Genetec (Korea), and the primer sequence is shown in Table 17. Lactoferrin was used as a positive control. The expression levels are shown in Table 18.

TGase-1 Primers Primer Sequense (5' to 3') SEQ ID NO Transglutaminse-1
(TGase-1) sense TGA ATA GTG ACA AGG TGT ACT GGC A 15 Transglutaminse-1
(TGase-1) antisense TTG TGA CAA TGA GTG TGC CGA T 16 β-actin sense GGC ACC CAG CAC AAT GAA G One β-actin antisense CCG ATC CAC ACG GAG TAC TTG 2

Test group Experimental concentration (μg/ml) TGase-1 mRNA
Expression rate (%) Control - 100.0 Lactoferrin 0.1 μM 155.8 Example 2
(Ginseng gel) 25 135.7 50 147.8 100 103.9

As a result of confirming the expression of the TGase-1 gene related to the skin barrier of the plant gel, it was confirmed that the expression of the gene in Example 2 was increased at various concentrations during treatment, thereby having the effect of strengthening the skin barrier.

2) Skin barrier Enzymes involved in formation Transglutaminase Evaluation of protein expression of -1 (Transglutaminase-1, TGase-1)

Epilife (gibco) medium containing human growth factor (gibco) and 3.5×10 ⁵ human keratin cells were inoculated into 100 mm dishes and cultured at 37° C. and 5% carbon dioxide for 24 hours. After 24 hours, the samples were treated in serum-free medium for each concentration and cultured for 3 to 5 days. Calcium chloride was used as a positive control for this experiment.

After the cells were scraped off using a scraper, the pellet was settled through centrifugation. Thereafter, a protein extraction solution was added and incubated, and the protein was quantified using a QubitTM fluorometer. After adding a loading buffer to the quantified protein, it was loaded on SDS-PAGE.

After transferring using the iBlot Dry Blotting system, blocking was performed. Thereafter, primary (TGase-1, β-actin) and secondary (GAM-HRP) antibodies were treated. After antibody treatment, west save cold ECL solution was treated, bands were photographed with chemi doc LuminoGraph II (ATTO), and the numerical values thereof were quantified and shown in Table 21.

Test group Experimental concentration (μg/ml) TGase-1/β-actin (%) Control - 100.0 Calcium chloride 1.8 mM 198.5 Example 4
(Bung full gel) 0.5 140.5 5 150.7

As a result of confirming the expression of the TGase-1 protein related to the skin barrier of the plant gel, it was confirmed that the expression of the protein in Example 4 was increased at various concentrations during treatment, thereby having the effect of strengthening the skin barrier.

3) Skin barrier One of the proteins that make up Involukrin ( Involucrin , INV ) Evaluation

Epilife (gibco) medium containing human growth factor (gibco) and 3.5×10 ⁵ human keratin cells were inoculated into 100 mm dishes and cultured at 37° C. and 5% carbon dioxide for 24 hours. After 24 hours, the samples were treated in serum-free medium for each concentration and cultured for 3 to 5 days. Calcium chloride was used as a positive control for this experiment.

After the cells were scraped off using a scraper, the pellet was settled through centrifugation. Thereafter, a protein extraction solution was added and incubated, and the protein was quantified using a QubitTM fluorometer. After adding a loading buffer to the quantified protein, it was loaded on SDS-PAGE.

After transferring using the iBlot Dry Blotting system, blocking was performed. Thereafter, primary (INV, β-actin) and secondary (GAM-HRP) antibodies were treated. After antibody treatment, west save cold ECL solution was treated, bands were photographed with chemi doc LuminoGraph II (ATTO), and the numerical values thereof were quantified and shown in Table 20.

Test group Experimental concentration (μg/ml) INV/β-actin (%) Control - 100.0 Calcium chloride 1.8 mM 160.4 Example 4
(Bung full gel) 0.5 144.2 5 145.0

When the expression of the INV protein related to the skin barrier of the plant gel was confirmed, it was confirmed that the expression of the skin barrier-related INV protein was increased in Example 4, as the protein expression was increased at various concentrations during treatment, thereby having the effect of strengthening the skin barrier.

Experimental Example 3: Evaluation of anti-skin aging efficacy of plant gel

Experimental Example 3-1: Investigation of the expression level of genes related to skin wrinkles of plant gel

1) MMP-1, 2, 9 expression reduction effect

Normal human fibroblasts were dispensed into a culture dish and cultured in an IMDM medium containing 10% bovine serum in an incubator at 37° C. and 5% CO ₂ for one day. The medium was replaced with HBSS (Hank's Balanced Salt Solution, Gibco, USA), irradiated with UVA 6 J/cm ² , replaced with IMDM containing no bovine serum, and then the sample was added and incubated at 37°C for 24 hours. QIAzol cells?? (Qiagen, USA) and proceed according to the manufacturer's method. After putting appropriate amount of a DEPC treated distilled water is dissolved Pellet the RNA to synthesize cDNA and then quantified using a Qubit ™ fluorometer (Invitrogen, USA) and ^{Qubit ⓡ RNA BR Assay Kit (Invitrogen} , USA) was performed Realtime PCR. For cDNA synthesis, a cDNA synthesis kit (qPCRBIO cDNA Synthesis kit, PCRBIOSYSTEMS, UK) was used, and an experiment was performed by the method of Kit. Realtime PCR was performed using the Realtime PCR Kit (2x qPCRBIO SyGreen Blue mix Lo-ROX, PCRBIOSYSTEMS, UK), and the experiment was performed by the method of the Kit, and after amplifying the gene using the Applied Biosystems 7500 FAST Real-Time PCR System The product was quantitatively analyzed. MMP-1, 2, 9, collagen type I and β-actin primers used in PCR were synthesized and used by Cosmo Genetech (Korea), and the sequence is shown in Table 21. The expression levels are shown in Tables 22, 23, 24, and 25. EGCG was used as a positive control.

Primers Primer Sequense (5' to 3') SEQ ID NO MMP-1 sense CCA CCT ATC ACT TCA GAT CAA CAA A 17 MMP-1 antisense TTC CTG TAG GCC AGT GTC AAA AT 18 MMP-2 sense GAT GCG GTA TAC GAG GCC C 19 MMP-2 antisense GAT CCA GTA TTC ATT CCC TGC AA 20 MMP-9 sense AAC ATC TTC GAC GCC ATC G 21 MMP-9 antisense AAT CGC CAG TAC TTC CCA TCC 22 collagen type I sense AGC AAG AAC CCC AAG GAC AA 23 collagen type I antisense CGA ACT GGA ATC CAT CGG TC 24 β-actin sense GGC ACC CAG CAC AAT GAA G One β-actin antisense CCG ATC CAC ACG GAG TAC TTG 2

MMP-1 mRNA expression level Test group Experimental concentration (μg/ml) MMP-1 mRNA expression rate (%) Non-irradiated group - 49.3 UVA irradiation group - 100.0 UVA + Example 1 (red ginseng gel) 100 89.8 250 81.8 500 56.3 UVA + Example 2 (ginseng gel) 25 80.4 50 70.6 100 50.1 UVA + Example 3 (rapeseed gel) 50 84.5 100 77.9 250 62.5 UVA + Example 4 (bottle paste gel) 25 58.0 50 49.8 UVA + Example 5 (camellia leaf gel) 25 78.4 50 68.4 UVA + Example 6 (Green Tea Leaf Gel) 2.5 86.8 UVA + Example 7 (Jeju Blue Bean Gel) 100 66.7 UVA + Example 8 (cheopseokgok gel) 100 64.0 UVA + Example 10 (Agar Gel) 50 46.6 100 76.1 250 81.1 UVA + EGCG 2.5 51.8

As a result of confirming the expression of the skin aging-related MMP-1 gene of the plant gel, Example 1, Example 2, Example 3, Example 4, Example 5, Example 6, Example 7, Example 8, Example 10 It was confirmed that there is an effect of skin aging as gene expression decreases at various concentrations during treatment.

MMP-2 mRNA expression level Test group Experimental concentration (μg/ml) MMP-2 mRNA expression rate (%) Non-irradiated group - 61.2 UVA irradiation group - 100.0 UVA + Example 1 (red ginseng gel) 100 84.3 250 74.6 500 69.7 UVA + Example 2 (ginseng gel) 25 (no effect) 92.2 50 68.0 100 73.2 UVA + EGCG 2.5 76.2

MMP-9 mRNA expression level Test group Experimental concentration (μg/ml) MMP-9 mRNA expression rate (%) Non-irradiated group - 66.6 UVA irradiation group - 100.0 UVA + Example 1 (red ginseng gel) 100 89.8 250 80.2 500 78.0 UVA + Example 2 (ginseng gel) 25 77.2 50 54.6 100 56.2 UVA + EGCG 2.5 79.7

As a result of confirming the expression of the skin aging-related MMP-9 gene of the plant gel, it was confirmed that the treatment of Examples 1 and 2 had skin aging effects as the gene expression decreased at various concentrations.

Collagen type I mRNA expression level Test group Experimental concentration (μg/ml) Col-I mRNA expression rate (%) Non-irradiated group - 100.0 UVA irradiation group - 78.0 UVA + Example 5 (camellia leaf gel) 25 99.3 50 97.3 UVA + Example 10 (Agar Gel) 50 100.7

As a result of confirming the expression of the skin aging-related Col-I gene of the plant gel, it was confirmed that the treatment of Example 5 and Example 10 had skin aging efficacy as gene expression was increased at various concentrations.

2) MMP-2, 9 enzyme activity inhibition effect

Normal human fibroblasts were dispensed into a culture dish and cultured in an IMDM medium containing 10% bovine serum in an incubator at 37° C. and 5% CO ₂ for one day. The medium was replaced with HBSS (Hank's Balanced Salt Solution, Gibco, USA), irradiated with UVA 6 J/cm ² , replaced with IMDM containing no bovine serum, and the sample was added and incubated at 37°C for 24 hours. The cell medium was collected with an Amicon® Ultra-4 centrifugal filter tube, and concentrated by centrifugation at 4°C, 3,000 rpm, 3-4 hours. After this, Qubit?? Quantification is performed using a fluorometer (Invitrogen, USA) and Qubit® protein BR Assay Kit (Invitrogen, USA). An appropriate amount of protein was separated using 10% gelatin zymogram gel, renaturated, and developed at 37°C. The gel was stained with coomassie blue solution, then replaced with destaining solution to check the shape of the band, and the band was photographed with Chemi doc LuminoGraphII (ATTO). The enzyme activity is shown in Tables 26 and 27.

Measurement of MMP-2 enzyme activity Test group Experimental concentration (μg/ml) MMP-2 Inactive form activity (%) MMP-2 Active form activity (%) Non-irradiated group - 84.3 76.7 UVA irradiation group - 100.0 100.0 UVA + Example 1 (red ginseng gel) 250 75.5 90.4 500 73.1 79.5 UVA + EGCG 2.5 76.8 24.2

When the expression of the MMP-2 protein related to skin aging of the plant gel was confirmed, it was confirmed that the treatment of Example 1 had skin aging efficacy as the protein expression decreased at various concentrations.

Measurement of MMP-9 enzyme activity Test group Experimental concentration (μg/ml) MMP-9 activity (%) Non-irradiated group - 33.6 UVA irradiation group - 100.0 UVA + Example 1 (red ginseng gel) 100 72.7 250 60.0 500 47.9 UVA + Example 2 (ginseng gel) 25 47.4 50 44.4 100 66.5 UVA + EGCG 2.5 84.4

When the expression of the MMP-9 protein related to skin aging of the plant gel was confirmed, it was confirmed that the treatment of Examples 1 and 2 had skin aging effects as the protein expression decreased at various concentrations.

Experimental Example 3-2: Investigation of expression levels of genes related to skin elasticity of plant gels

1) Collagen Type I, Elastin expression increase effect

Applied Biosystems 7500 FAST Real-Time PCR System (Applied Biosystems, CA, USA) was performed to measure the expression levels of Collagen Type I and Elastin genes. Human normal fibroblasts were cultured in an IMDM medium containing 10% bovine serum at a density of 1×10 ⁶ on a 60 mm plate in an incubator at 37° C. and 5% CO ₂ for one day. Thereafter, the medium was washed twice with HBSS (Hank's Balanced Salt Solution, Gibco, USA), and then replaced with IMDM containing no bovine serum, and the sample was added and incubated at 37°C for 24 hours. After incubation, the cells are collected with 1 ml of TRIzol (Invitrogen, USA) and transferred to a 1.5 ml tube, and 200 μl of Chloroform (Sigma, USA) is added, vortexed, and left at room temperature for 5 minutes. After that, centrifuge at 4°C and 15,000 rpm for 30 minutes, transfer the supernatant to a new tube, mix with the same amount of 2-propanol, stand at room temperature for 5 minutes, and centrifuge at 4°C and 15,000 rpm for 20 minutes. After centrifugation, discard 2-propanol, add 1 ml of 75% ethanol, centrifuge for 10 minutes at 4°C and 13,000 rpm, and dry the RNA pellet at room temperature to remove the remaining ethanol. After dissolving an appropriate amount of DEPC-treated purified water in a pellet, it is quantified using Qubit ^TM fluorometer (Invitrogen, USA) and Qubit ^® RNA BR Assay Kit (Invitrogen, USA), and then cDNA (complementary DNA) is synthesized for real-time PCR. Was carried out. For cDNA synthesis, a cDNA synthesis Kit (High Capacity RNA-to-cDNA Kit, Applied Biosystems, USA) was used, and an experiment was performed by the method of Kit. Real-time PCR was performed using a Real-time PCR Kit (2x qPCRBIO SyGreen Blue mix Lo-ROX, PCRBIOSYSTEMS, London, UK) and an experiment was performed by the method of the Kit, and Applied Biosystems 7500 FAST Real-Time PCR System was used. After amplifying the gene, the amplification product was quantitatively analyzed. Collagen Type I, Elastin and β-actin primers used in PCR were synthesized and used by Cosmo Genetech (Korea), and the sequence is shown in Table 28.

Primers Primer Sequense (5' to 3') SEQ ID NO collagen type I sense AGC AAG AAC CCC AAG GAC AA 23 collagen type I antisense CGA ACT GGA ATC CAT CGG TC 24 Elastin sense GGC CAT TCC TGG TGG AGT TCC 25 Elastin antisense AAC TGG CTT AAG AGG TTT GCC TCC A 26 β-actin sense GGC ACC CAG CAC AAT GAA G One β-actin antisense CCG ATC CAC ACG GAG TAC TTG 2

The reaction conditions for cDNA synthesis are Collagen Type I, Elastin, and β-actin in 95°C for 5 seconds, 60°C for 25 seconds and 40 cycles. Vitamin C, used as a control, is used as a substance that increases the expression of Collagen Type I and Elastin. The results of the expression amount are shown in Tables 29 and 30.

Collagen Type I mRNA expression level Test group Experimental concentration (μg/ml) Col-I mRNA expression rate (%) Control - 100.0 Vitamin C 100 117.7 Example 1
(Red Ginseng Gel) 100 120.9 250 119.9 500 140.7 Example 4
(Bung full gel) 50 120.5 100 132.7

When the expression of the Col-I gene related to skin elasticity of the plant gel was confirmed, it was confirmed that the treatment of Example 1 and Example 4 had a skin elasticity effect as Collagen synthesis increased at various concentrations.

Elastin mRNA expression level Test group Experimental concentration (μg/ml) Elastin mRNA expression rate (%) Control - 100.0 Vitamin C 100 122.2 Example 2
(Ginseng gel) 50 115.9 100 116.4 Example 3
(Oilseed gel) 50 157.0 100 113.8 250 117.1 Example 4
(Bung full gel) 25 173.9 Example 8
(Ironfigurine gel) 25 181.5 50 180.7 100 190.7

When the expression of the Elastin gene related to skin elasticity of the plant gel was confirmed, it was confirmed that the treatment of Example 3, Example 7, Example 8 increased the synthesis of Elastin at various concentrations, thereby having a skin elasticity effect.

Experimental Example 3-3: Skin photo-protection evaluation

After inoculation of normal human fibroblasts on the plate, the cells were cultured for one day in an incubator at 37°C and 5% CO ₂ . After that, the medium was washed with HBSS (Hank's Balanced Salt Solution, Gibco, USA), and then the sample was added and incubated at 37°C for 1 hour. UVB 50 mJ/cm ² was irradiated, replaced with a medium containing no bovine serum, and then a sample was added and cultured at 37°C for 24 hours. After removing the medium of the cells, the cells were washed with HBSS and replaced with a medium containing no bovine serum to which JC-1 dye was added, and cultured for 1 hour while maintaining the light-shielding state. After washing with HBSS, fluorescence was measured with an ELISA reader (Red: Ex=530 nm, Em=590 nm / Green: Ex=485 nm, Em=530 nm) and photographed with a fluorescence microscope.

Table 31 shows the measurement results.

Test group Experimental concentration (μg/ml) Red/green (%) Non-irradiated group - 100.0 UVB irradiation group - 73.9 UVB + Example 2 (ginseng gel) 25 102.2 50 99.4 100 102.9 UVB + Adenosine 7 86.1

As a result of the experiment, as shown in Table 31, it was shown that the cell function reduced by UVB irradiation during Example 2 treatment was significantly increased and recovered.

Experimental Example 3-4: Cell regeneration efficacy investigation

After inoculation of normal human fibroblasts on the plate, the cells were cultured for one day in an incubator at 37°C and 5% CO ₂ . Scar?? After scratching with Scratcher (SPL), washing with HBSS (Hank's Balanced Salt Solution, Gibco, USA) and replacing it with a medium containing no bovine serum, the samples of Example 4 (Bella gel) were 0, 5, 10, 20 μg/ ml was added and incubated at 37°C for 48 hours. After cultivation, a photograph of the cells was taken using a microscope (Leica). The results are shown in FIG. 1. As a result of the experiment, it was found that the damaged cells were regenerated compared to the control when 5 μg/ml was treated with Example 4 (Bungle gel).

Experimental Example 3-5: Measurement of cell function activation

After inoculation of normal human fibroblasts on the plate, the cells were cultured for one day in an incubator at 37°C and 5% CO ₂ . After that, the medium was washed with HBSS (Hank's Balanced Salt Solution, Gibco, USA) and then irradiated with UVB 50 mJ/cm ² , replaced with a medium containing no bovine serum, and then added to the sample at 37°C for 24 hours. Cultured. After removing the medium of the cells, the cells were washed with HBSS and replaced with a medium containing no bovine serum to which JC-1 dye was added, and cultured for 1 hour while maintaining the light-shielding state. After washing with HBSS, fluorescence was measured with an ELISA reader (Red: Ex=530 nm, Em=590 nm / Green: Ex=485 nm, Em=530 nm) and photographed with a fluorescence microscope. Table 32 shows the measurement results.

Test group Experimental concentration (μg/ml) Red/green (%) Non-irradiated group - 100.0 UVB irradiation group - 63.6 UVB + Example 4 (bottle full gel) 10 75.0 UVB + Adenosine 7 73.1

As a result of the experiment, as shown in Table 32, it was shown that the cell function reduced by UVB irradiation during the treatment of Example 4 increased by up to 11.4% and recovered.

Experimental Example 4: Evaluation of skin whitening efficacy of plant gel

Experimental Example 4-1: Melanin biosynthesis inhibition effect measurement

In order to measure the whitening effect of the plant gel, an experiment was performed by treating a sample in melanoma (B16-F1) cells of a mouse. DMEM (Dulbecco's modified Eagle's medium) medium containing 10% bovine serum and melanoma (B16-F1) cells were dispensed at a concentration of 1×10 ⁵ cells/well in a 6 well plate, and at 37°C and 5% CO ₂ for 24 hours. Cultured. After 24 hours, the sample is diluted by concentration in a new DMEM medium containing α-Melanocyte stimulate hormone (α-MSH) and 10% bovine serum and incubated for 72 hours. After 72 hours of incubation, cells from which all of the medium was removed were washed with 2 ml of PBS (phosphate buffer, saline) and treated with 0.5 ml of 1N NaOH to collect cells in a 1.5 ml tube to obtain intracellular melanin. The collected cells were transferred to a 96 well plate, and the absorbance was measured at 450 nm to determine the amount of melanin per protein. Control was used to treat 1,2-hexandiol used as a dissolution solvent, and α-MSH treatment group was used to treat both α-MSH and 1,2-hexandiol used as dissolution solvent. Arbutin (SK Bioland) was used as a positive control. The melanin biosynthesis inhibition test results are shown in Table 33.

Measurement of melanin production Test group Experimental concentration (μg/ml) Melanin formation rate per cell (%) Control 52.70 α-MSH treatment group 100.00 α-MSH + Example 7 (Jeju Blue Bean Gel) 50 85.11 α-MSH + Example 8 (cheopyrite gel) 50 78.76 100 72.76 250 76.89 α-MSH + Example 9 (Eam talc gel) 25 84.64 50 78.93 100 78.53 α-MSH + Arbutin 100 64.76

As a result of confirming the melanin production rate of the plant gel, it was confirmed that the treatment of Example 7, Example 8, and Example 9 had an inhibitory effect on melanin production by reducing the increased melanin production rate due to α-MSH.

Experimental example 4-2: the pathway for producing melanin melanogenesis relation Tyrosinase , TRP-1, TRP-2 mRNA expression level investigation

In order to measure the melanogenesis-related whitening effect of the plant gel, an experiment was conducted by treating a sample on melanoma (B16-F1) cells of a mouse. DMEM medium containing 10% bovine serum and melanoma (B16-F1) cells were dispensed at a concentration of 3X10 ⁵ cells/well in a 6 well plate and cultured at 37°C and 5% CO ₂ for 24 hours. After 24 hours, the sample is diluted by concentration in fresh DMEM medium containing α-MSH and 10% bovine serum and incubated for 24 hours. After culturing for 24 hours, the cells from which all the medium was removed were washed with 2 ml of PBS, and the cells were collected with QIAzol ^® (QIAGENL ^® , USA) to isolate RNA according to the manufacturer's method. The isolated RNA was quantified using a Qubit ^® fluoremeter with RNA BR Assay kit, and then cDNA was synthesized and real-time PCR was performed. For cDNA synthesis, a cDNA synthesis Kit (qPCRBIO cDNA Synthesis Kit, PCRBIOSYSTEMS, London, UK) was used, and an experiment was performed according to the method of Kit. For real-time PCR, a gene was amplified using a Real-time PCR Kit (2x qPCRBIO SyGreen Blue mix Lo-ROX, PCRBIOSYSTEMS, London, UK), and then the amplified product was quantitatively analyzed. Tyrosinase, Tyrosinase-related protein 1 (TRP-1) and Tyrosine-related protein 2 (TRP-2) and GAPDH primers used in PCR were synthesized and used by Cosmo Genetech (Korea), and the sequence is shown in Table 34 below. Done.

Primers Primer Sequense (5' to 3') SEQ ID NO Tyrosinase sense TAT AAA CTC AGT GTT TCC CTT TAT CAC AA 27 Tyrosinase antisense TCT ATG ATG ACA TGA AGT GGC AAA 28 TRP-1 sense GTT CAA TGG CCA GGT CAG GA 29 TRP-1 antisense CAA CGC AGC CAC TAC AGC A 30 TRP-2 sense TGG CTC ACT CCT TCC TGA ATG 31 TRP-2 antisense CAA ACA CAG GGT CGT TGG CT 32 GAPDH sense GGC ATC TTG GGC TAC ACT GAG 33 GAPDH antisense GGA AGA GTG GGA GTT GCT GTT G 34

The reaction conditions for cDNA synthesis were Tyrosinase, TRP-1 and TRP-2 and GAPDH at 95°C for 5 seconds, 60°C for 25 seconds, and 40 cycles. Control was used to treat 1,2-hexandiol used as a dissolution solvent, and α-MSH treatment group was used to treat both α-MSH and 1,2-hexandiol used as dissolution solvent. Arbutin, used as a positive control, is used as a substance that inhibits the expression of genes related to melanogenesis. The results of the expression levels are shown in Tables 35, 36, and 37.

Tyrosinase mRNA expression level Test group Conc. (μg/ml) Tyrosinase mRNA
Expression rate (%) Control - 68.37 α-MSH treatment group - 100.00 α-MSH + Example 8 (cheopyrite gel) 50 77.23 100 73.52 250 72.70 α-MSH + Example 9 (Eam talc gel) 25 76.10 50 60.65 100 73.57 α-MSH + Arbutin 100 83.45

TRP-1 mRNA expression level Test group Conc. (μg/ml) TRP-1 mRNA
Expression rate (%) Control 62.89 α-MSH treatment group 100.00 α-MSH + Example 8 (cheopyrite gel) 50 80.35 100 78.37 250 75.75 α-MSH + Example 9 (Eam talc gel) 25 85.52 50 63.03 100 80.18 α-MSH + Arbutin 100 86.97

As a result of confirming the expression of melanonogenesis-related genes in plant gels, it was found that the expression levels of the Tyrosinase and TRP-1 genes increased due to α-MSH during treatments in Examples 8 and 9 were decreased. Thus, the whitening effect by inhibition of melanonogenesis, a pathway for producing melanin. It was confirmed that there was.

Experimental example 4-3: Melanosomes Path of delivery melanosome transfer related PAR-2 mRNA Expression level investigation

To measure the whitening effect related to melanosome transfer of the plant gel, DMEM medium containing 10% bovine serum and HaCaT, an immortalized human keratinocyte, were dispensed at 2X10 ⁶ cells/well concentration in a 60 mm culture dish (corning) at 37°C for 24 hours. , Incubated in 5% CO ₂ conditions. After 24 hours, after washing twice with HBSS (Hank's Balanced Salt Solution, Gibco, USA), add HBSS and UVB 30 mJ/cm ² After irradiation, the samples are treated by concentration with DMEM medium containing no bovine serum and incubated in a 37°C, CO ₂ 5% incubator for 8 hours. After 8 hours of incubation, the cells from which all the medium was removed were washed with 2 ml of PBS, and the cells were collected with QIAzol® (QIAGENL®, USA) to isolate RNA according to the manufacturer's method. The isolated RNA was quantified using a Qubit® fluoremeter with RNA BR Assay kit, and then cDNA was synthesized and real-time PCR was performed. For cDNA synthesis, a cDNA synthesis Kit (qPCRBIO cDNA Synthesis Kit, PCRBIOSYSTEMS, London, UK) was used, and an experiment was performed according to the method of Kit. For real-time PCR, a gene was amplified using a Real-time PCR Kit (2x qPCRBIO SyGreen Blue mix Lo-ROX, PCRBIOSYSTEMS, London, UK), and then the amplified product was quantitatively analyzed. Protease activated receptor 2 (PAR-2) and β-actin primer used in PCR were synthesized and used by Cosmo Genetech (Korea), and the sequence is shown in Table 37 below.

Primers Primer Sequense (5' to 3') SEQ ID NO PAR-2 sense TGC TAG CAG CCT CTC TCT CC 35 PAR-2 antisense CCA GTG AGG ACA GAT GCA GA 36 β-actin sense GGC ACC CAG CAC AAT GAA G One β-actin antisense CCG ATC CAC ACG GAG TAC TTG 2

The reaction conditions for cDNA synthesis were 95°C for 5 seconds, 60°C for 25 seconds and 40 cycles for both PAR-2 and β-actin. Control was used to treat 1,2-hexandiol used as a dissolution solvent, and α-MSH treatment group was used to treat both α-MSH and 1,2-hexandiol used as dissolution solvent. EGCG, used as a positive control, is used as a substance that inhibits PAR-2 gene expression. Table 38 shows the results of the expression level.

PAR-2 mRNA expression level Test group Conc. (μg/ml) PAR-2 mRNA
Expression rate (%) Control - 51.01 UVB irradiation group - 100.00 UVB + Example 7 (Jeju Blue Bean Gel) 50 54.47 100 55.85 250 63.19 UVB + EGCG 3.5 66.26

As a result of confirming the expression of melanonosome transfer-related genes in the plant gel, it was found that the increased PAR-2 gene expression level was decreased due to UVB irradiation during treatment in Example 7, indicating that there is a whitening effect by inhibiting melanonosome transfer, a pathway for delivering melanosomes. Confirmed.

Formulation example

On the other hand, an example of the preparation of the cosmetic composition containing the plant gel of the present invention is described below. However, it is apparent to those skilled in the art that the following formulation examples are only intended to specifically describe the use examples of the present invention, and are not intended to limit the scope of the present invention to the following formulation examples.

Formulation example 1: Preparation of nourishing lotion (lotion)

As shown in Table 39 below, nutrient cosmetic water (lotion) containing the plant gels obtained in Examples 1 to 12 was prepared according to a conventional method.

number Raw material Content (% by weight) One Example 1-12 10.0 2 Sitosterol 1.7 3 Polygilyceryl 2-oleate 1.5 4 Ceteares-4 1.2 5 cholesterol 1.5 6 Dicetyl phosphate 0.4 7 Glycerol 5.0 8 Carboxyvinyl polymer 10.0 9 Sunflower oil 0.2 10 Xanthan gum 0.3 11 antiseptic a very small amount 12 Spices a very small amount 13 Purified water Balance

Formulation Example 2: Preparation of flexible lotion (skin)

As described in Table 40 below, a softening lotion (skin) containing the plant gels obtained in Examples 1 to 12 was prepared according to a conventional method.

number Raw material Content (% by weight) One Example 1-12 5.0 2 glycerin 3.0 3 Butylene glycol 2.0 4 Propylene glycol 2.0 5 Polyoxyethylene (60) hardened castor oil 1.0 6 ethanol 10.0 7 Preethanolamine 0.1 8 antiseptic a very small amount 9 Pigment a very small amount 10 Spices a very small amount 11 Purified water Balance

Formulation example 3: nutrition cream manufacturing

As described in Table 41 below, a nourishing cream containing the plant gels obtained in Examples 1 to 12 was prepared according to a conventional method.

number Raw material Content (% by weight) One Example 1-12 10.0 2 Sitosterol 4.0 3 Polygilyceryl 2-oleate 3.0 4 Ceteares-4 2.0 5 cholesterol 3.0 6 Dicetyl phosphate 0.4 7 Thick glycerol 5.0 8 Sunflower oil 22.0 9 Carboxyvinyl polymer 0.5 10 Triethanolamine 0.5 11 antiseptic a very small amount 12 Spices a very small amount 13 Purified water Balance

Formulation example 4: Preparation of essence

As described in Table 42 below, essences containing plant gels obtained in Examples 1 to 12 were prepared according to a conventional method.

number Raw material content ( weight% ) One Example 1-12 5.0 2 Sitosterol 1.7 3 Polygilyceryl 2-oleate 1.5 4 Ceteares-4 2.0 5 cholesterol 3.0 6 Dicetyl phosphate 0.4 7 Thick glycerol 5.0 8 Sunflower oil 22.0 9 Carboxyvinyl polymer 0.5 10 Triethanolamine 0.5 11 antiseptic a very small amount 12 Spices a very small amount 13 Purified water Balance

Formulation example 5: Preparation of foundation

As described in Table 43 below, a foundation containing the plant gels obtained in Examples 1 to 12 was prepared according to a conventional method.

number Raw material content ( weight% ) One Example 1-12 1.0 2 Beeswax 2.0 3 Cychromechone 2.0 4 Floating paraffin 5.0 5 Squalane 5.0 6 Stearic acid 2.0 7 Lipophilic glycerin monostearate 3.0 8 Caprylic/Capric Triglyceride 4.0 9 glycerin 4.0 10 Propylene glycol 3.0 11 Butylene glycol 3.0 12 Triethanolamine 1.0 13 Aluminum Magnesium Silicate 0.5 14 Pigment 12 15 antiseptic a very small amount 16 Spices a very small amount 17 Purified water Balance

Formulation example 6: Hair conditioner Produce

As described in Table 44 below, hair conditioners containing the plant gels obtained in Examples 1 to 12 were prepared according to a conventional method.

number Raw material content ( weight% ) One Example 1-12 5.0 2 ethanol 3.5 3 glycerin 1.5 4 Propylene glycol 2.5 5 Stearyl triethine ammonium chloride 2.0 6 Pigment a very small amount 7 Spices a very small amount 8 Purified water Balance

<110> SK bioland Co., Ltd. <120> Use of Gel Comprising Plant As Raw Material <130> PN190183 <160> 36 <170> KoPatentIn 3.0 <210> 1 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> beta-actin sense primer <400> 1 ggcacccagc acaatgaag 19 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> beta-actin antisense primer <400> 2 ccgatccaca cggagtactt g 21 <210> 3 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Aquaporin-3(AQP-3) sense primer <400> 3 gggaccagtc ggaagggat 19 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Aquaporin-3(AQP-3) antisense primer <400> 4 cacagatgga caggctgcct 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Hyaluronic acid-2 (HAS-2) sense primer <400> 5 acaacacctt agttcctcta 20 <210> 6 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Hyaluronic acid-2 (HAS-2) antisense primer <400> 6 agcagtgata tgtctcct 18 <210> 7 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> CD-44 sense primer <400> 7 gctattgaaa gccttgcaga g 21 <210> 8 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> CD-44 antisense primer <400> 8 cgcagatcga tttgaatata acc 23 <210> 9 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Pro filaggrin (Pro FLG) sense primer <400> 9 ctgatggtat tcaagttggc t 21 <210> 10 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Pro filaggrin (Pro FLG) antisense primer <400> 10 actgtgcttt ctgtgcttgt 20 <210> 11 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> Occludin(OCDN) sense primer <400> 11 ccacctatca cttcagatca acaaa 25 <210> 12 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Occludin(OCDN) antisense primer <400> 12 ttcctgtagg ccagtgtcaa aat 23 <210> 13 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Serine Palmitoyl Transferase (SPT) sense primer <400> 13 tggtcatttg gcccaggtc 19 <210> 14 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Serine Palmitoyl Transferase (SPT) antisense primer <400> 14 tcccaaccat tggcttcaca tc 22 <210> 15 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> Transglutaminse-1(TGase-1) sense primer <400> 15 tgaatagtga caaggtgtac tggca 25 <210> 16 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Transglutaminse-1(TGase-1) antisense primer <400> 16 ttgtgacaat gagtgtgccg at 22 <210> 17 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> MMP-1 sense primer <400> 17 ccacctatca cttcagatca acaaa 25 <210> 18 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> MMP-1 antisense primer <400> 18 ttcctgtagg ccagtgtcaa aat 23 <210> 19 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> MMP-2 sense primer <400> 19 gatgcggtat acgaggccc 19 <210> 20 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> MMP-2 antisense primer <400> 20 gatccagtat tcattccctg caa 23 <210> 21 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> MMP-9 sense primer <400> 21 aacatcttcg acgccatcg 19 <210> 22 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> MMP-9 antisense primer <400> 22 aatcgccagt acttcccatc c 21 <210> 23 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> collagen type I sense primer <400> 23 agcaagaacc ccaaggacaa 20 <210> 24 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> collagen type I antisense primer <400> 24 cgaactggaa tccatcggtc 20 <210> 25 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Elastin sense primer <400> 25 ggccattcct ggtggagttc c 21 <210> 26 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> Elastin antisense primer <400> 26 aactggctta agaggtttgc ctcca 25 <210> 27 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> Tyrosinase sense primer <400> 27 tataaactca gtgtttccct ttatcacaa 29 <210> 28 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Tyrosinase antisense primer <400> 28 tctatgatga catgaagtgg caaa 24 <210> 29 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TRP-1 sense primer <400> 29 gttcaatggc caggtcagga 20 <210> 30 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> TRP-1 antisense primer <400> 30 caacgcagcc actacagca 19 <210> 31 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TRP-2 sense primer <400> 31 tggctcactc cttcctgaat g 21 <210> 32 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TRP-2 antisense primer <400> 32 caaacacagg gtcgttggct 20 <210> 33 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> GAPDH sense primer <400> 33 ggcatcttgg gctacactga g 21 <210> 34 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> GAPDH antisense primer <400> 34 ggaagagtgg gagttgctgt tg 22 <210> 35 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PAR-2 sense primer <400> 35 tgctagcagc ctctctctcc 20 <210> 36 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PAR-2 antisense primer <400> 36 ccagtgagga cagatgcaga 20

Claims (17)

Plant gel cosmetic composition comprising an oxidized fine pulverized plant powder and a residual amount of a solvent.
The plant gel cosmetic composition of claim 1, wherein the composition comprises powder of oxidized finely pulverized land plants.
The method of claim 2, wherein the land plant is an outpost, root, stem, or blood (leaf) of a land plant, and the composition is 1-25% (w/v) of oxidized microparticles based on the total weight of the composition. The plant gel cosmetic composition comprising the pulverized plant powder.
The method of claim 3, wherein the outposts, roots, stems, or their blood (baked) of the land plant are selected from the group consisting of red ginseng, ginseng, centella and their blood (bak), and the composition is for skin moisturizing. , Plant gel cosmetic composition.
The method of claim 3, wherein the outposts, roots, stems, or their blood (baked) of the land plant is selected from the group consisting of ginseng, red ginseng, centella, and their blood (baked), and the composition strengthens skin barriers. Yongin, plant gel cosmetic composition.
The method of claim 3, wherein the outpost, root, stem, or blood (bak) of the land plant is selected from the group consisting of red ginseng, ginseng, rapeseed, centella asiatica, cheopseokgok, and blood (bak) thereof, and the The composition is for preventing skin aging, a plant gel cosmetic composition.
The plant according to claim 3, wherein the outposts, roots, stems, or blood (bak) of the land plants are selected from the group consisting of ginseng, centella, and their blood (bak), and the composition is for UV protection. Gel cosmetic composition.
The plant gel cosmetic composition according to claim 3, wherein the outposts, roots, stems, or blood (leaf) of the terrestrial plants are Cheolpiseokgok, and the composition is for skin whitening.
The method of claim 2, wherein the land plant is a leaf, a fruit, a seed, or the blood of a land plant, and the composition is 5-30% (w/v) of oxidized microparticles based on the total weight of the composition. The plant gel cosmetic composition comprising the pulverized plant powder.
The plant gel cosmetic composition according to claim 9, wherein the leaves, fruits, seeds, or blood (bak) of the land plant are camellia leaves, green tea leaves, or a combination thereof, and the composition is for skin moisturizing.
The method of claim 9, wherein the leaves, fruits, seeds, or blood (bak) of the land plant are selected from the group consisting of camellia leaves, green tea leaves, Jeju green beans, and blood (bak) thereof, and the composition is Plant gel cosmetic composition for preventing skin aging.
The plant gel cosmetic composition according to claim 9, wherein the leaves, fruits, seeds, or blood (bak) of the land plant are Jeju blue beans, and the composition is for skin whitening.
The plant gel cosmetic composition according to claim 1, wherein the composition comprises powder of oxidized finely pulverized marine plants.
The plant gel cosmetic composition according to claim 13, wherein the marine plant contains 1-20% (w/v) of oxidized finely pulverized plant powder based on the total weight of the composition.
The plant gel cosmetic composition of claim 14, wherein the marine plant is an agar, and the composition is for skin moisturizing.
The plant gel cosmetic composition of claim 14, wherein the marine plant is an agar, and the composition is for preventing skin aging.
The plant gel cosmetic composition of claim 14, wherein the marine plant is Ecklonia cava, and the composition is for skin whitening.

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