JP2015134737A - External preparation for skin - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an external preparation for skin containing an active ingredient derived from natural product, having an excellent biosafety and a skin-beautifying or whitening effect which keeps or may improve the skin in a youthful and healthy condition, and protecting the skin from external environmental factors, such as ultraviolet rays .SOLUTION: An external preparation for skin comprising an extract which is derived from Camellia sinensis (L.) O. Kuntze and has a barrier function improving effect, an anti-oxidant effect, and an anti-inflammatory effect of the skin, exhibits a rough and dry skin improving effect, a preventive and improving effect of skin aging (wrinkle, sagging, and the like), an improving effect of skin tension and gloss, a protective effect of the skin from oxidation damage due to external factors (ultraviolet rays and chemical substances) and inflammation, and a preventive and improving effect for stains, freckles, chloasma, and the like.

Description

  The present invention relates to a skin external preparation blended component obtained from a plant belonging to the camellia family and having excellent skin physiological activity and biosafety, and a skin external preparation formulated with such a component.

  Skin aging is induced by internal factors such as inactivation of cell growth / differentiation with age, decrease in hormone secretion, quantitative decrease in extracellular matrix components, sunlight (ultraviolet rays), exhaust gas, etc. It is a phenomenon that is caused by complex intertwining of external factors such as cell / tissue damage caused by active oxygen or inflammation.

For example, ultraviolet rays and chemical substances (nitrogen compounds, sulfur compounds, etc.) contained in exhaust gas cause oxidative damage to the skin and alter biological components, resulting in generation of antigens in the skin. For this reason, external factors such as ultraviolet rays and chemical substances cause skin inflammation and allergies due to antigens.

In addition, the skin originally has a barrier function to retain moisture and a function to recover oxidative damage,
When their functions are reduced due to external factors such as ultraviolet rays, the amount of water held by the skin is lost through each layer, which causes rough skin, pigmentation such as spots and freckles, and wrinkles that are skin aging phenomena. , Tarmi, etc. occur.

  In order to prevent rough skin and aging of the skin, and to keep the skin healthy and youthful, the use of various active ingredients has been proposed in the past, and these active ingredients (skin-beautifying ingredients, whitening ingredients, etc.) were formulated. An external preparation for skin is on the market. For example, vitamin C, vitamin E, antioxidants such as superoxide dismutase (hereinafter abbreviated as SOD); anti-inflammatory agents such as glycyrrhizic acid; various ultraviolet absorbers; α-hydroxycarboxylic acid, placenta extract, γ Cell activation components such as amino-β-hydroxybutyric acid; extracellular matrix components such as collagen, elastin, hyaluronic acid; humectants such as urea. In addition, kojic acid is known as a substance that suppresses the occurrence of pigmentation such as skin spots, buckwheat and liver spots, and is widely used as an active ingredient of a whitening agent. However, the above antioxidants, moisturizers, whitening agents, and the like have problems in terms of skin safety, and there are demands for components for external preparations for skin that are excellent in skin safety.

  In view of the problems of the prior art, the present inventors have intensively studied to find new skin-beautifying and whitening components derived from natural products from the viewpoint of skin safety. As a result, the extract of Benifumi, a plant belonging to the genus Camellia belonging to the camellia family, has an excellent barrier function improving action, antioxidant action and anti-inflammatory action, and thus it was excellent by blending the extract. It has been found that it is possible to provide an external preparation for skin that exhibits a skin beautifying effect and a whitening effect, and is excellent in skin safety.

  Conventionally, the use of tea extracts belonging to the genus Camellia as a skin external preparation is disclosed in Patent Documents 1 and 2, and epigallocatechin contained in tea such as Benifumi Takashi has antiallergic or anti-allergic properties. Patent Document 3 discloses that it has an inflammatory action.

Japanese Unexamined Patent Publication No. 01-128933 Japanese Patent Laid-Open No. 10-072361 JP 2000-159670 A

However, the extract of Benifumi has excellent barrier function improving effect, antioxidant effect and anti-inflammatory effect, and from their synergistic effect, it improves skin aging phenomenon such as rough skin, wrinkles, tarmi, skin firmness, It has not been known to improve gloss and to have an effect of improving spots and freckles.

The present invention is an external preparation for skin containing as an active ingredient an extract of Camellia sinensis (L.) O. Kuntze, a plant belonging to the genus Camellia.

The present invention is an external preparation for skin comprising as an active ingredient an extract of Benifumi, which is a plant belonging to the genus Camellia, which is an epidermal cell keratinization promoting effect, an antioxidant effect (inhibition of intracellular active oxygen generation, intracellular SOD activity enhancement action) and anti-inflammatory effect (prostaglandin E2 (hereinafter referred to as “PGE ₂ ”) production inhibitory action), and the synergistic effect thereof improves the skin barrier function, and wrinkles, tarmi Improves skin aging, etc., and improves skin firmness and gloss, and also helps prevent and improve skin damage (oxidation damage and inflammation damage) due to external factors such as ultraviolet rays and the like. An external preparation for skin can be provided.

FIG. 1 is a diagram showing the epidermal cell keratinization promoting effect of the red rich extract according to the present invention. FIG. 2 is a diagram showing the epidermal cell keratinization promoting effect of the red rich extract according to the present invention. FIG. 3 is a diagram showing the epidermal cell keratinization promoting effect of the control. FIG. 4 is a diagram showing the effect of suppressing the generation of active oxygen in fibroblasts by ultraviolet rays of the red rich extract according to the present invention. FIG. 5 is a diagram showing the intracellular SOD activity enhancement effect of the red rich extract according to the present invention. FIG. 6 is a diagram showing the PGE ₂ production inhibitory effect of the red rich extract according to the present invention.

Hereinafter, preferred embodiments of the present invention will be described in detail.
The extraction material used in the present invention is Camellia sinensis (L.) O. Kuntze, a plant belonging to the genus Camellia.

  The material used in the present invention may be any of Benifumi's whole plants, leaves, flower parts, stems, seeds, fruits, roots, etc., but the use of whole plants or leaves is preferred.

  For the preparation of the extract, first, Benifumi (for example, whole grass, leaves, etc.) is washed with water in advance if necessary to remove foreign substances, and then dried or crushed or crushed as necessary. Extraction is performed in contact with an extraction solvent. In addition, regarding the drying method, what processed common tea leaves like what is called green tea, black tea, oolong tea, etc. may be used.

  As an extraction solvent, water; lower alcohols such as methanol, ethanol, propanol; polyhydric alcohols such as ethylene glycol, propylene glycol, 1,3-butylene glycol, glycerin; ethyl acetate, butyl acetate, methyl propionate, etc. Esters; Ketones such as acetone and methyl ethyl ketone; Ethers such as ethyl ether and isopropyl ether; Hydrocarbon solvents such as n-hexane, toluene and chloroform, and the like. These may be used alone or in combination of two or more. Used.

Among these extraction solvents, the skin extract keratinization promoting effect, antioxidant effect (inhibitory action on the generation of intracellular active oxygen, action to enhance intracellular SOD activity) and anti-inflammatory effect (inhibition action on PGE ₂ production) of the obtained extract, Furthermore, from the viewpoint of skin irritation and from the viewpoint that it can be widely applied to external preparations for the skin, hydrophilic solvents such as water, lower alcohols or polyhydric alcohols are preferred in the present invention. is there. Preferable examples of using this hydrophilic solvent include, for example, water, lower alcohols (especially ethanol), or polyhydric alcohols (especially 1,3-butylene glycol) alone, or water and lower alcohols. Examples include the use of a mixed solvent with (especially ethanol) or a mixed solvent of water and a polyhydric alcohol (especially 1,3-butylene glycol, glycerin). Among them, water alone or water with 1,3 -A mixed solvent of butylene glycol is particularly preferred.

  When the mixed solvent is used, for example, if the mixed solvent is water and 1,3-butylene glycol, the volume ratio (hereinafter the same) is 1:10 to 20: 1, and the mixed solvent is water and ethanol. If it exists, it is preferable to set it as the range of 1: 10-20: 1 if it is a mixed solvent of 1: 10-25: 1 and water and glycerol.

  Moreover, the weight ratio of the dried portion (for example, whole grass or leaf) of Benifumi and the extraction solvent is preferably in the range of 1: 1 to 1:50, more preferably in the range of 1: 5 to 1:35. It is.

  In preparing the extract, the pH is not particularly limited, but it is generally preferably in the range of 3-9. In this sense, if necessary, the extraction solvent is blended with an alkaline adjusting agent such as sodium hydroxide, sodium carbonate or potassium hydroxide, or an acidic adjusting agent such as citric acid, hydrochloric acid, phosphoric acid or sulfuric acid. You may adjust so that it may become pH of.

  Extraction conditions such as extraction temperature and extraction time vary depending on the type and pH of the solvent used. For example, immersion using water or 1,3-butylene glycol or a mixture of water and 1,3-butylene glycol as a solvent. In the case of the process, the extraction temperature is generally in the range of 1 to 90 ° C., preferably 40 ° C. to 80 ° C., and the extraction time is 0.5 to 24 hours in the case of 40 ° C., preferably The range is 0.5 to 6 hours.

Prior to the extraction process of the present invention or in parallel with the extraction process, Benifumi may be subjected to a hydrolysis process as necessary. This may improve the storage stability and the like of the extract of Benifumi and may use the extract as a skin external preparation compounding agent more effectively.

In the case of subjecting the extract to an enzymatic hydrolysis treatment, examples of the enzyme include proteolytic enzymes such as actinase, papain and pepsin, starch degrading enzymes such as glucoamylase, α-amylase and β-amylase, cellulase, hemicellulase and pectinase. One or two or more selected from the group of enzymes of lipolytic enzymes such as fibrinolytic enzymes and lipases may be used, but one or more selected from these enzyme groups, respectively. It is more preferable to use a combination of these enzymes.

  The amount of the enzyme added is, for example, in the range of 0.01 to 10% by weight, more preferably, in the range of 0.01 to 10% by weight based on the solid content of the whole plant or flower part of Benifumi. It is in the range of 1 to 2.0% by weight.

  In general, the extract prepared as described above may be adjusted to a pH of 3 to 8 and used as it is as a skin external preparation compound as it is, or as a desired concentration by vacuum concentration or the like. May be used. In addition, the extract may be dried by a conventional method such as spray drying.

  In addition, the extract prepared as described above may be refrigerated for a certain period of time in order to enhance storage stability and the like, and the supernatant may be used as a skin external preparation compounding agent.

  Examples of skin external preparations (cosmetics, quasi-drugs, etc.) containing the extract of Benifumi of the present invention include emulsions, creams, lotions, essences, packs, lipsticks, foundations, liquid foundations, makeup press powders, cheeks. White powder, facial cleansers, body shampoos, hair shampoos, soaps, and the like, and hair growth agents, and further bath agents, but of course are not limited thereto.

  The blended amount of Benifumi extract in the external preparation for skin of the present invention is generally 0.002 to 1.0% by weight, preferably 0.02 to 0 in the case of a basic external preparation for the solid content of the extract. In the case of an external preparation for makeup skin, generally in the range of 0.002 to 1.0% by weight, preferably in the range of 0.02 to 0.2% by weight. In the case of an external preparation for cleaning, In general, it is in the range of 0.002 to 10.0% by weight, preferably 0.02 to 7.0% by weight.

  The external preparation for skin of the present invention includes, in addition to the essential extract of Benifumi, components that are usually used in external preparations for skin, such as oil components, surfactants (synthetic systems, natural products), moisturizers, A viscosity agent, an emulsifier or an emulsification aid, an antiseptic / bactericidal agent, a powder component, an ultraviolet absorber, an antioxidant, a pigment, a fragrance, other physiologically active components, and the like can be appropriately blended as necessary. In addition, as long as the effectiveness and features of the extract of Benifumi noki according to the present invention are not impaired, it may be combined with other physiologically active ingredients in a topical skin preparation.

Here, as the oil component, for example, olive oil, jojoba oil, castor oil, soybean oil, rice oil, rice germ oil, palm oil, palm oil, cacao oil, meadow foam oil, sheer butter, tea tree oil, avocado oil, Oils derived from plants such as macadamia nut oil and plant-derived squalane; Fats derived from animals such as mink oil and turtle oil; waxes such as beeswax, carnauba wax, rice wax, lanolin; liquid paraffin, petrolatum, paraffin wax, squalane, etc. Hydrocarbons; fatty acids such as myristic acid, palmitic acid, stearic acid, oleic acid, isostearic acid, cis-11-eicosenoic acid; higher alcohols such as lauryl alcohol, cetanol, stearyl alcohol; isopropyl myristate, palmitic acid Isopropyl, me Butyl phosphate, 2-ethylhexyl glycerides, higher fatty acid octyldodecyl (octyl stearate dodecyl and the like), and the synthetic esters and synthetic triglycerides such like.

Examples of the surfactant include polyoxyethylene alkyl ether, polyoxyethylene fatty acid ester, polyoxyethylene sorbitan fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, polyoxyethylene glycerin fatty acid ester, polyoxyethylene hydrogenated castor oil, polyoxyethylene Nonionic surfactants such as oxyethylene sorbitol fatty acid esters; fatty acid salts, alkyl sulfates, alkylbenzene sulfonates, polyoxyethylene alkyl ether sulfates, polyoxyethylene fatty amine sulfates, polyoxyethylene alkyl phenyl ether sulfates, Polyoxyethylene alkyl ether phosphates, α-sulfonated fatty acid alkyl ester salts, polyoxyethylene alkyl phenyl ether phosphates, Quaternary ammonium salt, primary to tertiary fatty amine salt, trialkylbenzylammonium salt, alkylpyridinium salt, 2-alkyl-1-alkyl-1-hydroxyethylimidazolinium salt, N N, N-dimethyl-N-alkyl-N-carboxymethylammoniobetaine, N, N, N-trialkyl-N-, N, N-dimethyl-N-alkyl-N-carboxymethylammoniobetaine Amphoteric surfactants such as alkylene ammoniocarboxybetaine and N-acylamidopropyl-N ′, N′-dimethyl-N′-β-hydroxypropylammoniosulfobetaine can be used.

Examples of emulsifiers and emulsifiers include stevia derivatives such as enzyme-treated stevia, lecithin and its derivatives (hydrogenated lecithin, etc.), lactic acid bacteria fermented rice, lactic acid bacteria fermented rice, lactic acid bacteria fermented cereals (wheat, beans, millet, etc.) It can also be blended.

Examples of the humectant include glycerin, propylene glycol, dipropylene glycol, 1,3-butylene glycol, polyethylene glycol, sorbitol, xylitol, sodium pyrrolidone carboxylate, and sugars such as trehalose, mucopolysaccharides (for example, hyaluron). Acid and derivatives thereof, chondroitin and derivatives thereof, heparin and derivatives thereof, elastin and derivatives thereof, collagen and derivatives thereof, NMF related substances, lactic acid, urea, higher fatty acid octyldodecyl, seaweed extract, silane root (white and white) Examples include extracts, various amino acids, and derivatives thereof.

Examples of the thickener include components derived from brown algae, green algae or red algae such as alginic acid, agar, carrageenan and fucoidan; silane root (white) extract; polysaccharides such as pectin, locust bean gum and aloe polysaccharide; xanthan gum, Gums such as tragacanth gum and guar gum; cellulose derivatives such as carboxymethylcellulose and hydroxyethylcellulose; synthetic polymers such as polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, acrylic acid / methacrylic acid copolymer; hyaluronic acid and its derivatives; Examples include glutamic acid and its derivatives.

Examples of the antiseptic / bactericidal agent include urea; paraoxybenzoates such as methyl paraoxybenzoate, ethyl paraoxybenzoate, propyl paraoxybenzoate, and butyl paraoxybenzoate; phenoxyethanol, dichlorophene, hexachlorophene, chlorhexidine hydrochloride, benzaza chloride Luconium, salicylic acid, ethanol, undecylenic acid, phenols, jamal (imidazodenyl urea), 1,2-pentanediol, various essential oils, bark dry matter, and the like.

Examples of powder components include sericite, titanium oxide, talc, kaolin, bentonite, zinc oxide, magnesium carbonate, magnesium oxide, zirconium oxide, barium sulfate, silicic anhydride, mica, nylon powder, polyethylene powder, silk powder, and cellulose. System powders, powders of cereals (rice, wheat, corn, millet, etc.), powders of beans (soybeans, red beans, etc.).

Examples of the ultraviolet absorber include ethyl paraaminobenzoate, ethylhexyl paradimethylaminobenzoate, amyl salicylate and derivatives thereof, 2-ethylhexyl paramethoxycinnamate, octyl cinnamate, oxybenzone, 2,4-dihydroxybenzophenone, 2-hydroxy-4 -Methoxybenzophenone-5-sulfonate, 4-tertiarybutyl-4-methoxybenzoylmethane, 2- (2-hydroxy-5-methylphenyl) benzotriazole, urocanic acid, ethyl urocanate, aloe extract, etc. .

Antioxidants include, for example, butylhydroxyanisole, butylhydroxytoluene, propyl gallate, vitamin E and its derivatives (eg, vitamin E nicotinate, vitamin E linoleate, etc.), murasakixikib extract, peony extract, silane root (white and white) ) There are extracts, etc.

Examples of whitening agents include t-cycloamino acid derivatives, kojic acid and derivatives thereof, ascorbic acid and derivatives thereof, hydroquinone or derivatives thereof, ellagic acid and derivatives thereof, nicotinic acid and derivatives thereof, resorcinol derivatives, tranexamic acid and derivatives thereof, 4 -Methoxysalicylic acid potassium salt, magnolignan (5,5'-dipropyl-biphenyl-2,2'-diol), 4-HPB (rhodenol, 4- (4-hydroxyphenyl) -4-butanol)), hydroxybenzoic acid And derivatives thereof, vitamin E and derivatives thereof, α-hydroxy acid, and AMP (adenosine monophosphate, adenosine monophosphate). These may be used alone or in combination.

  Examples of the kojic acid derivatives include kojic acid esters such as kojic acid monobutyrate, kojic acid monocaprate, kojic acid monopalmitate, kojic acid dibutyrate, kojic acid ethers, kojic acid sugar derivatives such as kojic acid glucoside, etc. However, as the ascorbic acid derivatives, for example, L-ascorbic acid-2-phosphate sodium, L-ascorbic acid-2-phosphate magnesium, L-ascorbic acid-2-sulfate sodium, L-ascorbic acid-2 -Ascorbic acid ester salts such as magnesium sulfate, L-ascorbic acid-2-glucoside, ascorbic acid sugar derivatives such as L-ascorbic acid-5-glucoside, 6-position acylated products of these ascorbic acid sugar derivatives (acyl group is Hexanoyl group, o L-ascorbic acid tetrafatty acid esters such as L-ascorbic acid tetraisopalmitate, L-ascorbic acid tetralaurate, 3-O-ethylascorbic acid, L-ascorbic acid- 2-phosphate-6-O-palmitate sodium, glyceryl ascorbic acid or acylated derivatives thereof, ascorbyl glycerol derivatives such as bisglyceryl ascorbic acid, as hydroquinone derivatives, arbutin (hydroquinone-β-D-glucopyranoside), α-Arbutin (hydroquinone-α-D-glucopyranoside) and the like as tranexamic acid derivatives include tranexamic acid esters (for example, tranexamic acid lauryl ester, tranexamic acid hexadecyl ester, tranet And amides of tranexamic acid (for example, tranexamic acid methylamide). Resorcinol derivatives include, for example, 4-n-butylresorcinol, 4-isoamylresorcinol, and the like. Examples of the -dihydroxybenzoic acid derivative include 2,5-diacetoxybenzoic acid, 2-acetoxy-5-hydroxybenzoic acid, and 2-hydroxy-5-propionyloxybenzoic acid. Examples of the nicotinic acid derivative include nicotinic acid amide. Examples of the α-hydroxy acid such as benzyl nicotinate include lactic acid, malic acid, succinic acid, citric acid, and α-hydroxyoctanoic acid.

As a physiologically active ingredient, as a whitening ingredient, for example, placenta extract, Sakuhakuhi extract, yukinoshita extract, perilla extract, rice bran extract or hydrolyzate thereof, white coconut extract or hydrolyzate thereof, white coconut Fermented product, damask rose extract, peony extract or hydrolyzate thereof, lactic acid bacteria fermented rice, murasaki kib extract, lotus seed extract or hydrolyzate thereof, lotus seed fermented product, banquet extract, pearl hydrolyzate, pearl barley Fermented seeds, fermented royal jelly, fermented sake lees, Pandanus amaryllifolius Roxb. Extract, Arcangelicia flava Merrilli extract, chamomile extract, and other anti-aging ingredients Coral grass extract, rice leaf extract or hydrolyzate thereof, eggplant (water eggplant, long eggplant, Kamo eggplant, rice eggplant ) Extracts or hydrolysates thereof, extracts of apricot fruits, extracts of seaweeds such as catamen giraffes, extracts of marine flowering plants such as sea cucumbers, fermented soymilk, jellyfish water, rice extract or hydrolysates thereof , Fermented rice extract, germinated rice extract or hydrolyzate thereof, fermented germinated rice, black bean extract or hydrolyzate thereof, linoleic acid and its derivatives or processed products (eg liposomal linoleic acid), animal or fish origin Collagen and derivatives thereof, elastin and derivatives thereof, glycyrrhizic acid and derivatives thereof (dipotassium salt, etc.), t-cycloamino acid derivatives, vitamin A and derivatives thereof, allantoin, diisopropylamine dichloroacetate, γ-amino-β-hydroxybutyric acid, Gentian extract, licorice extract, carrot extract, aloe extract, beetroot extract , Luffa extract, Ulva pertusa extract, Juazeiro (Zizyphus joazeiro) have extracts.

  Next, the present invention will be described more specifically with reference to production examples, formulation examples, and test examples, but the present invention is not limited thereto. In the following, all parts are parts by weight, and all percentages are% by weight.

Production Example 1 Preparation of Benifumi's extract 1200 g of purified water was added to 80 g of Benifumi (dried tea leaves), extracted at 50 ° C. for 3 hours, filtered, and then 1000 g of brown clear beech extract (solid content 1.6) %).

Production Example 2 Preparation of the extract of Benifumi Takashi (added 1200 g of 50% 1,3-butylene glycol aqueous solution to 80 g of Benifumi (dried tea leaves), followed by extraction at 50 ° C. for 3 hours, followed by filtration and 850 g of a brown transparent Benifuku extract (Solid content concentration 1.3%) was obtained.

Formulation Example 1 Lotion [A component] part olive oil 1.0
Polyoxyethylene (5.5) cetyl alcohol 5.0
Butylparaben 0.1
[Component B] Extract solution of Part Production Example 1 5.0
Ethanol 5.0
Glycerin 5.0
1,3-butylene glycol 5.0
Potassium hydroxide appropriate amount Purified water Amount that makes the total amount 100 parts [component C]
Fragrance Appropriate amounts of component A and component B were each heated to 80 ° C. or higher, component B was added to component A and stirred, and then homogenized for 2 minutes with Hiscotron (5000 rpm). After cooling this to 50 degreeC, C component was added and stirred and mixed, and also it cooled to 30 degrees C or less, and the lotion was obtained.

Formulation Example 2 A lotion was obtained in the same manner as in Formulation Example 1 except that 5.0 parts of the extract solution in Production Example 2 was used instead of the extract solution in Production Example 1 contained in the B component of Formulation 1. .

Formulation Example 3 Emulsion [Component A] Part Liquid paraffin 6.0
Hexalan 4.0
Jojoba oil 1.0
Polyoxyethylene (20) sorbitan monostearate 2.0
Soy lecithin 1.5
[Component B] Part Extract solution of Production Example 1 3.0
L-ascorbic acid-2-glucoside 2.0
Potassium hydroxide 0.5
Glycerin 3.0
1,3-butylene glycol 2.0
Carboxymethylcellulose 0.3
Sodium hyaluronate 0.01
Purified water Amount of 100 parts [component C]
Perfume
The components A and B were each heated to 80 ° C. or higher and then mixed by stirring. After cooling this to 50 ° C., component C was added and further stirred and mixed to obtain an emulsion.

Formulation Example 4 Emulsion Emulsion in the same manner as in Formulation Example 3 except that 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 parts of potassium hydroxide are used instead of 2.0 parts of L-ascorbic acid-2-glucoside in Formulation Example 3 Got.

Formulation Example 5 Emulsion [Component A] Part Liquid paraffin 6.0
Hexalan 4.0
Jojoba oil 1.0
Polyoxyethylene (20) sorbitan monostearate 2.0
Soy lecithin 1.5
[Component B] Part Extract solution of Production Example 1 5.0
L-ascorbic acid-2-glucoside 2.0
Potassium hydroxide 0.5
Arbutin 3.0
Glycerin 3.0
1,3-butylene glycol 2.0
Carboxymethylcellulose 0.3
Sodium hyaluronate 0.01
Amount of purified water totaling 100 parts

Formulation Example 6 Extraction solution of lotion [component] part production example 2 10.0
Ethanol 10.0
Glycerin 3.0
1,3-butylene glycol 2.0
Methylparaben 0.2
Citric acid 0.1
Sodium citrate 0.3
Carboxyvinyl polymer 0.1
Fragrance Appropriate amount Potassium hydroxide Appropriate amount Purified water An amount that makes the total amount 100 parts.

Formulation Example 7 Essence [ingredient] part ethanol 2.0
Glycerin 5.0
1,3-butylene glycol 5.0
Methylparaben 0.1
Hyaluronic acid 0.1
Extract solution of Production Example 1 5.0
Citric acid 0.3
Sodium citrate 0.6
Purified water After hyaluronic acid was dissolved in purified water in an amount of 100 parts, the remaining raw materials were sequentially added and dissolved by stirring to obtain a transparent essence.

Formulation Example 8 Essence was obtained in the same manner as in Formulation Example 7, except that 5.0 parts of the extract solution of Production Example 2 was used instead of the extract solution of Production Example 1 in the components of Essence Formulation Example 7.

Example 9 Liquid foundation [component A] part stearic acid 2.4
Propylene glycol monostearate 2.0
Cetostearyl alcohol 0.2
Liquid lanolin 2.0
Liquid paraffin 3.0
Isopropyl myristate 8.5
Propylparaben 0.05
[Component B] Extract solution of Part Production Example 1 5.0
Sodium carboxymethylcellulose 0.2
Bentonite 0.5
Propylene glycol 4.0
Triethanolamine 1.1
Methylparaben 0.1
Purified water Total amount of 100 parts [component C] parts Titanium oxide 8.0
Talc 4.0
Coloring pigment Appropriate amounts The above-mentioned components A and B were heated and mixed and stirred. This was reheated, the above C component was added, poured into a mold, and stirred until it reached room temperature to obtain a liquid foundation.

Formulation Example 10 Body shampoo [component A] part N-lauroylmethylalanine sodium 25.0
Palm oil fatty acid potassium liquid (40%) 26.0
Palm oil fatty acid diethanolamide 3.0
Methylparaben 0.1
[Component B] Part Extract solution of Production Example 2 5.0
1,3-butylene glycol 2.0
Purified water Amount to be 100 parts A component and B component are each heated to 80 ° C and dissolved uniformly, then B component is added to A component and stirring is continued to cool to room temperature to obtain a body shampoo It was.

Test Example 1 Epidermal cell keratinization promoting effect Normal human epidermal cells were seeded in a 96-well microplate at a concentration of 1 × 10 ⁴ cells / well. As a medium, a growth additive-containing medium Humdia KG2 (Kurabo) was used. After pre-culture at 37 ° C. for 1 day, the extract of Production Example 1 is added to the medium so as to have a concentration of 0.25% and 0.5% as the sample solution (final concentration as the sample solution with respect to the total amount of the medium) And further cultured at 37 ° C. for 1 day. Next, the morphology of the cells was observed with a phase contrast microscope (OLYMPAS), and the progress of keratinization was determined based on the degree of flatness of the cells. Further, as a control, the same operation was performed using a PBS (−) solution instead of the sample solution, and the cell morphology was observed.

The results of Test Example 1 are shown in FIGS.
FIG. 1 is a photomicrograph showing the morphology of epidermal cells cultured with the extract (0.25%) of Production Example 1 added, and FIG. 2 shows the addition of the extract (0.5%) of Production Example 1 FIG. 3 is a photomicrograph showing the morphology of epidermal cells cultured with the addition of a PBS (−) solution as a control. As shown in FIG. 1 to FIG. 3, it was confirmed that the extract of Production Example 1 of the present invention promotes epidermal cell flattening, that is, differentiation of epidermal cells in a concentration-dependent manner as compared with the control. .

Test Example 2 Inhibition of intracellular active oxygen generation by ultraviolet rays of fibroblasts Human dermis-derived fibroblasts NB1RGB were seeded in a 96-well plate at 2 × 10 ⁴ cells / well. As the medium, Eagle medium containing 10% NCS was used. After pre-culturing for 1 day under conditions of 37 ° C. and 5.0% CO ₂ , the extract of Production Example 1 has a concentration of 0.5% as a sample solution (final concentration as a sample solution with respect to the total amount of the medium). And then further cultured for 1 day under the same conditions. Next, the medium was removed and washed with 200 μL of Hanks Balanced Salts Solution (HBSS (+)), and then 100 μL of HBSS (+) and 100 μL of 10 μM 2 ′, 7′-Dichlorodihydrofluorine dicatenate were added. After incubating at 37 ° C. for 15 minutes, the supernatant was removed, washed with 200 μL of HBSS (+), 200 μL of HBSS (+) was added, and UV-B (Philips TL20W / 12RS) 50 mJ / cm ² was irradiated. did. Immediately after UV-B irradiation, 100 μL of 1% Triton-X-containing HBSS (+) was added to disrupt the cells, and the fluorescence intensity of this reaction solution at Ex 485 nm / Em 530 nm was measured. In addition, in the case of no sample (Control) added with PBS (−) instead of the sample solution, the same operation as described above was performed, and the activity of the sample addition group with respect to the amount of generated active oxygen (100) was obtained. The relative value of oxygen generation was determined. Further, a UV-B non-irradiated section was provided, PBS (-) was added instead of the sample solution, and the same operation was performed to determine a relative value for the amount of active oxygen generated by Control (100).

The results of Test Example 2 are shown in FIG. As shown in FIG. 4, it was confirmed that the extract of Production Example 1 according to the present invention has an excellent inhibitory effect on the generation of intracellular active oxygen.

Test Example 3 Effect of enhancing SOD activity in fibroblasts Human dermis-derived fibroblasts NB1RGB were seeded in a petri dish at 6 × 10 ⁵ cells / pet. As the medium, Eagle medium containing 10% NCS was used. After pre-incubation for 1 day under conditions of 37 ° C. and 5.0% CO ₂ , PBS (−) or the extract solution of Production Example 1 was used as a sample solution at a concentration of 0.5% (as a sample solution relative to the total amount of the medium). To the final concentration) and further cultured for 2 days under the same conditions. Next, the medium was removed, washed with 1 mL of PBS (−), 400 μL of 0.1% Triton-X, 1 mM PMSF-containing PBS (−) was added to disrupt the cells, and the crude enzyme solution was prepared. . 0.2 M Tris-HCl buffer 50 μL, 1 mM ethylenediaminetetraacetic acid (EDTA) disodium solution 20 μL, 0.75 mM nitroblue tetrazolium chloride (NBT) solution 10 μL, 1 mM xanthine solution 20 μL, 0.06 U / mL xanthine oxidase solution 50 μL, crude A sample solution was prepared by mixing 50 μL of the enzyme solution. This sample solution was incubated at 37 ° C. for 3 minutes to generate superoxide. Then, the absorbance at 570 nm was measured for the amount of formazan produced by reducing NBT with superoxide. The amount of protein in each crude enzyme solution was measured by the Bradford method, and the amount of formazan produced per amount of protein was calculated. In addition, the same operation was carried out using 0.1% Triton-X, 1 mM PMSF-containing PBS (-) instead of the crude enzyme solution. The value was determined and used as the NTB reduction rate (%), that is, the superoxide residual rate (%).

The result of Test Example 3 is shown in FIG. As shown in FIG. 5, it was confirmed that the extract of Production Example 1 according to the present invention has an excellent effect of enhancing SOD activity in fibroblasts.

Test Example 4 PGE ₂ production inhibitory effect test The anti-inflammatory effect of the extract of Benifumi according to the present invention was evaluated by a PEG ₂ production inhibitory test. Here, PGE ₂ is an inflammatory chemical mediator that causes skin inflammation, and is secreted in skin cells damaged by external factors such as ultraviolet rays and chemical substances. Therefore, in the present invention, the anti-inflammatory effect was evaluated by this PGE ₂ production inhibitory effect test.

<Experiment method>
Usaki cornea-derived cells (SIRC) are suspended in Eagle's minimum essential medium containing 10% FBS, seeded 5.0 × 10 ^{3 in} 96-well plates, cultured at 37 ° C. for 3 days, and then produced in the culture solution. The extract solution (sample solution) of Example 1 was added and further cultured for 24 hours. Here, the extract solution of Production Example 1 was added so that the final concentration as a solution with respect to the total amount of the culture solution was 2.5%. In addition, a 50% 1,3-butylene glycol aqueous solution was added in place of the sample solution and cultured for 24 hours to serve as a control. Next, 100 mJ / cm ² ultraviolet B wave was irradiated from the bottom of the incubator, and after further culturing for 2 days, the amount of PGE ₂ secreted into the culture supernatant was measured using a PGE ₂ measurement kit (manufactured by Cayman Caymic). And measured. Then, the relative value of the PGE ₂ amount at the time of adding the sample solution was determined by setting the PGE ₂ amount at the time of adding the control to 100.

The results of Test Example 4 are shown in FIG. As shown in FIG. 6, it was confirmed that the extract of Production Example 1 according to the present invention remarkably suppresses the production of PGE ₂ promoted by ultraviolet (UV) irradiation.

Claims (1)

An external preparation for skin containing an extract of Camellia sinensis (L.) O. Kuntze, a plant belonging to the genus Camellia.