JP5773356B2 - Cosmetics - Google Patents

Description

  The present invention relates to a cosmetic composition obtained from a salamander belonging to the genus Citrus, and having excellent skin physiological activity and improved biosafety, and a cosmetic comprising such a composition.

Conventionally, various natural products derived anti-aging components (antioxidant components, moisturizing components, etc.) have been studied, and it has been reported that yam extract, also known as herbal medicine, has an anti-aging effect. Here, “Yamabuchi” means Zanthoxylum piperitum, a plant belonging to the genus Rutaceae, Zanthoxylum, and its seeds and pericarp are anti-aging (antioxidant, inhibition of collagenase activity). It has been reported that an extract of salamander can be used as a cosmetic ingredient (Patent Documents 1 to 3).
JP 2001-354958 A JP 2003-342123 A JP 2003-342184 A

  As described above, the use of a salamander extract as a cosmetic compounding agent has been studied because it has an anti-aging effect. However, since a salmon extract has a high skin irritation potential, As a result, there was a problem in terms of safety. Furthermore, there is a problem that the extract of salamander does not exhibit a sufficient anti-aging effect. In addition, the salamander extract also has a problem of stability that coloration and starch develop over time.

  As a result of intensive studies in view of the problems of the prior art, the present inventors have significantly reduced skin irritation and improved stability by hydrolyzing salamander extracts with enzymes. In addition, a new activity to enhance the activity of ceramide synthase (β-glucocerebrosidase) in epidermal cells has been created, and it has a significantly superior epidermal cell activation effect compared to the extract. The headline and the present invention were completed.

The present invention is a cosmetic comprising as an active ingredient an enzyme hydrolyzate obtained by hydrolyzing an extract of a salamander (Zanthoxylum piperitum), which is a plant of the genus Rutaceae, Zanthoxylum, with an enzyme. is there.
In the present invention, the enzyme is a proteolytic enzyme.
In the present invention, the enzyme is a combination of a proteolytic enzyme and a lipolytic enzyme.
In the present invention, the enzyme hydrolyzate is obtained by hydrolyzing a salamander seed extract with an enzyme.

The present invention is a cosmetic comprising, as an active ingredient, an enzyme hydrolyzate obtained by hydrolyzing an extract of a salamander, which is a plant belonging to the genus Citrus, with the enzyme hydrolyzate, Compared with enzyme-treated salamander extract, skin irritation is remarkably suppressed and stability is improved, so there is no risk of erythema when applied to the skin, and storage The occurrence of coloring and starch in the inside can also be suppressed, and a cosmetic with high biological safety and storage stability can be provided.

Furthermore, the enzyme hydrolyzate of a salamander extract, which is an active ingredient of the present invention, has a ceramide synthase activity enhancing action not found in an enzyme-untreated salamander extract, and also has an effect of activating epidermal cells. Since it is remarkably enhanced compared to the extract, the cosmetics containing the enzyme hydrolyzate have excellent skin firmness, gloss improvement, and skin aging phenomena such as wrinkles and tarmi. Prevents and improves.
In the present specification, the term cosmetic is used in a broad sense including so-called cosmetics and quasi-drugs.

Hereinafter, the present invention will be described in detail.
The term “sauna” used in the present invention is a salamander (Zanthoxylum piperitum) that is a plant of the genus Rutaceae (Zanthoxylum).

  For the preparation of the enzyme hydrolyzate of the salamander extract of the present invention, an appropriate portion such as a salamander seed, seed coat, fruit, fruit skin, flower, leaf, stem, branch, bark, flower, whole plant, preferably If necessary, the seeds are washed with water in advance to remove foreign substances, and after being dried or dried, they are chopped or pulverized as necessary, such as a dipping method, countercurrent method, or supercritical extraction method. It can be performed by contacting with an extraction solvent according to a conventional method.

  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, water, a mixed solvent of water and lower alcohols (especially ethanol), from the viewpoint of the effectiveness of the extract solution obtained and the ease and efficiency of transfer to enzymatic hydrolysis treatment, or The use of a mixed solvent of water and a polyhydric alcohol (especially 1,3-butylene glycol or propylene glycol) can be mentioned, and the use of water alone is most preferable.

  The mixing ratio in the case of using a mixed solvent is, for example, 100: 1 to 1: 1 in a volume ratio (hereinafter the same) if the mixed solvent is water and ethanol, and 100: if the mixed solvent is water and glycerin. If it is 1-1: 4 and it is a mixed solvent of water and 1,3-butylene glycol, it is preferable to set it as the range of 100: 1 to 1: 2.

  In preparing the extract, the pH is not particularly limited, but it is generally preferably in the range of 2-8. 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.

  Extraction conditions such as extraction temperature and extraction time vary depending on the type and pH of the solvent used. For example, when water is used as the extraction solvent, the extraction temperature is generally 4 to 90 ° C, preferably 20 to 60 ° C. The extraction time is generally 1 to 8 hours, preferably 2 to 6 hours.

  Next, the extract solution obtained here is subjected to an enzymatic hydrolysis treatment. Here, when a solvent other than water (a mixed solvent of water and ethanol or water and 1,3-butylene glycol) is used for the preparation of the extract solution, the extract solvent is once removed from the extract and obtained here. The resulting extract is redissolved in water and then subjected to enzymatic degradation.

  Examples of the enzyme used for the hydrolysis treatment include Neulase F3G (Amano Enzyme Co., Ltd. product), which is known to have protease activity and lipase activity. In addition, proteolytic enzymes such as actinase, pepsin, papain, chymotrypsin, carboxypeptidase, aminopeptidase, bromelain, amylolytic enzymes such as glucoamylase and α-amylase, fibrinolytic enzymes such as cellulase, hemicellulase and pectinase, and lipase One or two or more selected from lipolytic enzymes such as these may be used, and it is particularly preferable to use one or more enzymes selected from these four enzyme groups in combination. Among these, it is most preferable to use a proteolytic enzyme and a lipolytic enzyme in combination.

  Hydrolysis with an enzyme is performed by adding one or more of the above enzymes to a salamander extract solution and performing an enzyme reaction under conditions near the optimum pH and temperature of the enzyme used. To be implemented. When two or more kinds of enzymes are used in combination, two or more kinds of enzymes may be allowed to act simultaneously according to the characteristics of the enzyme used, or may be allowed to act sequentially without changing or changing the reaction conditions. .

The amount of the enzyme used is preferably in the range of 0.001 to 50 parts by weight, more preferably 0.1 to 10 parts by weight per one enzyme with respect to 100 parts by weight of the solid content of the salamander extract solution. Part range. The enzyme treatment time varies depending on the type of enzyme used and the like, but is generally in the range of 0.5 to 24 hours, preferably in the range of 1 to 6 hours. The enzyme treatment described above may be performed simultaneously with the extraction treatment, which is the previous step, depending on the case.

After the treatment with the enzyme, the enzyme is deactivated by using an appropriate method such as a method of heating the enzyme treatment solution to 80 ° C. or higher to obtain an enzyme hydrolyzate solution.

In general, the enzyme hydrolyzate solution obtained as described above is adjusted to a pH of 2 to 8 and then mixed with cosmetics as it is or, if necessary, to a predetermined concentration by vacuum concentration or the like. Blended into cosmetics. In some cases, the powder may be pulverized according to a conventional method such as spray drying or freeze drying.

  The enzyme hydrolyzate of the present invention prepared as described above has an excellent ceramide synthase enhancement effect and epidermis as shown in the test examples to be described later, which is less irritating to the skin and excellent in biological safety. Since it has a cell activation effect, the cosmetic containing the hydrolyzate can improve the firmness and gloss of the skin, and can prevent and improve the occurrence of wrinkles and sagging.

  Examples of cosmetics containing the hydrolyzate of the present invention include basic cosmetics such as emulsions, creams, lotions, essences, and packs, and makeup makeups such as lipsticks, foundations, liquid foundations, makeup press powders, cheeks, and white powders. And cosmetics, facial cleansers, body shampoos, soaps and other cleaning cosmetics, and bath agents, but of course are not limited thereto.

  The blending amount of the enzyme hydrolyzate in the cosmetic of the present invention is generally 0.002 to 1.0% by weight, preferably 0.02 to 0 in the case of a basic cosmetic, as the solid content of the enzyme hydrolyzate. In the case of makeup cosmetics, generally in the range of 0.002 to 1.0% by weight, preferably in the range of 0.02 to 0.2% by weight. It is 0.002 to 10.0% by weight, preferably 0.02 to 7.0% by weight.

  In addition to the enzyme hydrolyzate that is an essential component, the cosmetics of the present invention include components usually used in cosmetics, such as oily components, surfactants (synthetic and natural products), moisturizers, thickeners, An antiseptic / bactericidal agent, a powder component, an ultraviolet absorber, an antioxidant, a pigment, a fragrance and the like can be appropriately blended as necessary. In addition, as long as the effectiveness and features of the enzyme hydrolyzate of the present invention are not impaired, it may be combined with other physiologically active ingredients in cosmetics.

  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.
In addition, as emulsifiers or emulsifiers, stevia derivatives such as enzyme-treated stevia, lecithin and its derivatives, lactic acid bacteria fermented rice, lactic acid bacteria fermented rice, lactic acid bacteria fermented cereals (wheat, legumes, miscellaneous cereals, etc.), juteiro (Rhamnaceae zizyphus joazeiro) An extract or the like can also be blended.

  Examples of humectants include glycerin, propylene glycol, dipropylene glycol, 1,3-butylene glycol, polyethylene glycol, sorbitol, xylitol, sodium pyrrolidonecarboxylate, and sugars such as trehalose, lactic acid bacteria fermented rice, and mucopolysaccharides. (For example, hyaluronic acid and its derivatives, chondroitin and its derivatives, heparin and its derivatives, etc.), elastin and its derivatives, collagen and its derivatives, NMF related substances, lactic acid, urea, higher fatty acid octyldodecyl, seaweed extract, beech extract Products, seafood-derived collagen and derivatives thereof, various amino acids and derivatives thereof.

  Examples of the thickener include brown algae, green algae or red algae-derived components such as alginic acid, agar, carrageenan and fucoidan; beech extract; polysaccharides such as pectin, locust bean gum and aloe polysaccharide; xanthan gum, tragacanth gum, guar gum and the like Gums; Cellulose derivatives such as carboxymethylcellulose and hydroxyethylcellulose; Synthetic polymers such as polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer and acrylic acid / methacrylic acid copolymer; Hyaluronic acid and its derivatives; Polyglutamic acid and its derivatives Is mentioned.

  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, beech extract, rice extract and the like.

  Examples of physiologically active ingredients include whitening ingredients such as t-cycloamino acid derivatives, kojic acid and derivatives thereof, ascorbic acid and derivatives thereof, hydroquinone derivatives, ellagic acid and derivatives thereof, resorcinol derivatives, tranexamic acid and derivatives thereof, 4-methoxy Salicylic acid potassium salt, magnolignan (5,5′-dipropyl-biphenyl-2,2′-diol), 4-HPB (rhodenol, 4- (4-hydroxyphenyl) -4-butanol)), AMP (adenosine monophos) Fate, adenosine monophosphate), placenta extract, nicotinic acid and its derivatives, Sakuhakuhi extract, yukinoshita extract, rice bran extract, rice bran extract hydrolyzate, lactic acid bacteria fermented rice, lactic acid bacteria fermented rice, lactic acid bacteria fermented cereals ( Wheat, beans, cereals), white coconut extract, white coconut hydrolyzed extract Murasakixikib extract, fermented lotus seed, peony extract, party extract, fermented pearl barley, fermented royal jelly, fermented sake lees, Pandanus amaryllifolius Roxb. Extract, Arcangelicia flava (Arcangelicia flava) flava Merrilli) extract, chamomile extract (trade name: Chamomile ET), seaweed extract such as kombu, seaweed extract such as sea cucumber, linoleic acid and its derivatives or processed products (eg liposomal linoleic acid), 2,5-dihydroxybenzoic acid derivatives, etc. are also used as skin aging preventing / beautifying components, such as collagen derived from animals or fish and derivatives thereof, elastin and derivatives thereof, nicotinic acid and derivatives thereof, glycyrrhizic acid and derivatives thereof (dipotassium salt) Etc.), t-cycloamino acid derivatives, vitamin A and its derivatives, bi Mineral E and its derivatives, allantoin, α-hydroxy acids, diisopropylamine dichloroacetate, γ-amino-β-hydroxybutyric acid, gentian extract, licorice extract, pearl extract, carrot extract, aloe extract, etc. , Rice extract hydrolyzate, rice bran extract hydrolyzate, rice fermentation extract, honey comb extract, anaaaosa extract, seaweed extract, etc. is there.

  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 (2-O-α-D-glucopyranosyl-L-ascorbic acid), L-ascorbic acid-5-glucoside (5-O-α) -D-glucopyranosyl-L-ascorbine Acid) ascorbic acid sugar derivatives, acylated 6-positions of these ascorbic acid sugar derivatives (acyl groups are hexanoyl, octanoyl, decanoyl, etc.), L-ascorbic acid tetraisopalmitate, L-ascorbic acid tetra L-ascorbic acid tetrafatty acid esters such as lauric acid ester, 3-O-ethylascorbic acid, L-ascorbic acid-2-phosphate-6-O-palmitate sodium and the like are hydroquinone derivatives such as arbutin (hydroquinone). -Β-D-glucopyranoside), α-arbutin (hydroquinone-α-D-glucopyranoside) and the like, and as resorcinol derivatives, for example, 4-n-butylresorcinol, 4-isoamylresorcinol and the like are 2,5-dihydroxybenzoic acid. Derivatives and For example, 2,5-diacetoxybenzoic acid, 2-acetoxy-5-hydroxybenzoic acid, 2-hydroxy-5-propionyloxybenzoic acid, etc., and nicotinic acid derivatives include, for example, nicotinic acid amide, nicotinic acid benzyl, etc. However, examples of the vitamin E derivative include vitamin E nicotinate and vitamin E linoleate, and examples of the α-hydroxy acid include lactic acid, malic acid, succinic acid, citric acid, and α-hydroxyoctanoic acid.

  Next, the present invention will be described more specifically with reference to production examples, 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 enzyme hydrolyzate from salamander seeds (1)
After drying and pulverizing the seeds of the citrus salamander, 60 g of the pulverized product is mixed with 600 g of purified water, and then 0.6 g of Neulase F3G (Amano Enzyme Co., Ltd. product) is added and the mixture is heated at 40 ° C. for 4 hours After extraction, the mixture was filtered, and then heated at 80 ° C. for 1 hour to deactivate the enzyme, followed by filtration to obtain 420 g of a brown transparent enzyme hydrolyzate solution (solid content concentration: 0.69%).

Production Example 2 Preparation of enzyme hydrolyzate from salamander seeds (2)
After drying and pulverizing the seeds of the citrus salamander, 60 g of the pulverized product is mixed with 600 g of purified water, 0.3 g of actinase and 0.3 g of lipase are added, and extraction is performed at 40 ° C. for 4 hours. Then, the mixture was filtered and then heated at 80 ° C. for 1 hour to deactivate the enzyme, followed by filtration to obtain 415 g of a brown transparent enzyme hydrolyzate solution (solid content concentration 0.68%).

Production Example 3 Preparation of enzyme hydrolyzate from salamander seeds (3)
The seeds of Citrus salamander were dried and pulverized, and after mixing 600 g of purified water with 60 g of the pulverized product, 0.3 g of papain and 0.3 g of lipase were added, and extraction was performed at 40 ° C. for 4 hours. After filtration, the mixture was heated at 80 ° C. for 1 hour to inactivate the enzyme, followed by filtration to obtain 418 g of a brown transparent enzyme hydrolyzate (solid content concentration: 0.69%).

Production Example 4 Preparation of Enzyme Hydrolyzate from Salamander Seeds (4)
After drying and pulverizing the seeds of Citrus salamander, 60 g of the pulverized product was mixed with 600 g of purified water, 0.3 g of actinase and 0.3 g of papain were added, and extraction was performed at 40 ° C. for 4 hours. Then, the mixture was filtered, and then heated at 80 ° C. for 1 hour to deactivate the enzyme, followed by filtration to obtain 425 g of a brown transparent enzyme hydrolyzate solution (solid content concentration 0.68%).

Production Example 5 Preparation of enzyme hydrolyzate of salamander seeds (5)
The seeds of the mandarin family Salamander are dried and pulverized, and after mixing 600 g of purified water with 60 g of the pulverized product, 0.6 g of actinase is added, extraction is performed at 40 ° C. for 4 hours, and then filtered. The enzyme was inactivated by heating at 80 ° C. for 1 hour, followed by filtration to obtain 420 g of a brown transparent enzyme hydrolyzate solution (solid content concentration 0.67%).

Production Example 6 Preparation of enzyme hydrolyzate from salamander seeds (6)
The seeds of the citrus salamander were dried and pulverized, and 60 g of the pulverized product was mixed with 600 g of purified water, followed by extraction at 40 ° C. for 3 hours, followed by filtration to obtain an extract solution. To this extracted solution, 0.6 g of Neulase F3G (Amano Enzyme Co., Ltd. product) is added, hydrolase treatment is performed at 40 ° C. for 3 hours, and then heated at 80 ° C. for 1 hour to deactivate the enzyme After filtration, 418 g of a brown transparent enzyme hydrolyzate solution was obtained (solid content concentration 0.67%).

Comparative Production Example 1 Preparation of salamander seed extract (1)
The seeds of Citrus salamander are dried and pulverized, and 60 g of the pulverized product is mixed with 600 g of purified water, followed by extraction at 40 ° C. for 4 hours, followed by filtration to obtain a brown transparent salamander seed extract. 425 g of a solution was obtained (solid content concentration 0.65%).

Example 1. Cream [Component A] Liquid paraffin 5.0
Hexalan (Note 1) 4.0
Paraffin 5.0
Glyceryl monostearate 2.0
Polyoxyethylene (20) sorbitan monostearate 6.0
Butylparaben 0.1
(Note 1) Technoble Co., Ltd. glyceryl trioctanoate
[B component]
Enzyme hydrolyzate of Production Example 1 5.0
Glycerin 5.0
Carboxymethyl monostearate 0.1
Moiston C (Note 2) 1.0
Amount of purified water 100 parts (Note 2) NMF component manufactured by Technoble Co., Ltd.
[C component]
Perfume
The components A and B were each heated to 80 ° C. or higher and then mixed by stirring. After this was cooled to 50 ° C., component C was added and further stirred and mixed to obtain a cream.

Example 2 Cream A cream was obtained in the same manner as in Example 1 except that the enzyme hydrolyzate of Production Example 2 was added instead of the enzyme hydrolyzate of Production Example 1 in [Component B] of Example 1.

Example 3 Cream A cream was obtained in the same manner as in Example 1 except that the enzyme hydrolyzate of Production Example 3 was added instead of the enzyme hydrolyzate of Production Example 1 in [Component B] of Example 1.

Example 4 Cream A cream was obtained in the same manner as in Example 1 except that the enzyme hydrolyzate of Production Example 4 was added instead of the enzyme hydrolyzate of Production Example 1 in [Component B] of Example 1.

Example 5 FIG. Cream A cream was obtained in the same manner as in Example 1 except that the enzyme hydrolyzate of Production Example 5 was added instead of the enzyme hydrolyzate of Production Example 1 in [Component B] of Example 1.

Example 6 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
Methylparaben 0.15
Ethylparaben 0.03
[B component]
Enzyme hydrolyzate of Production Example 1 5.0
Glycerin 3.0
1,3-butylene glycol 2.0
Carboxymethylcellulose 0.3
Sodium hyaluronate 0.01
Amount of purified water totaling 100 parts
[C component]
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.

Example 7 Emulsion An emulsion was obtained in the same manner as in Example 6 except that the enzyme hydrolyzate of Production Example 2 was added instead of the enzyme hydrolyzate of Production Example 1 in [Component B] of Example 6.

Example 8 FIG. Emulsion An emulsion was obtained in the same manner as in Example 6 except that the enzyme hydrolyzate of Production Example 3 was added instead of the enzyme hydrolyzate of Production Example 1 in [Component B] of Example 6.

Example 9 Emulsion An emulsion was obtained in the same manner as in Example 6 except that the enzyme hydrolyzate of Production Example 4 was added instead of the enzyme hydrolyzate of Production Example 1 in [Component B] of Example 6.

Example 10 Emulsion An emulsion was obtained in the same manner as in Example 6 except that the enzyme hydrolyzate of Production Example 5 was added instead of the enzyme hydrolyzate of Production Example 1 in [Component B] of Example 6.

Example 11 Lotion [component A] part Enzyme hydrolyzate of Production Example 1 5.0
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
Perfume
Potassium hydroxide appropriate amount Purified water Amount that makes 100 parts in total
The above ingredients were mixed to obtain a lotion.

Example 12 Lotion [A component] part olive oil 1.0
Polyoxyethylene (5.5) cetyl alcohol 5.0
Butylparaben 0.1
[B component]
Enzyme hydrolyzate of Production Example 1 5.0
Ethanol 5.0
Glycerin 5.0
1,3-butylene glycol 5.0
Methylparaben 0.1
Potassium hydroxide appropriate amount Purified water Amount that makes 100 parts in total
[C component]
Fragrance
Appropriate amount
After each component A and component B was heated to 80 ° C. or higher, the component B was added to the component A and stirred, and further homogenized with Hiscotron (5000 rpm) for 2 minutes. 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.

Example 13 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
Methylparaben 0.15
Ethylparaben 0.03
[B component]
Enzyme hydrolyzate of Production Example 1 5.0
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
Amount of purified water totaling 100 parts
[C component]
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.

Example 14 Latex In addition to using 2.0 parts of L-ascorbic acid-2-phosphate magnesium in place of 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 parts of potassium hydroxide in the component B of Example 13 Obtained an emulsion in the same manner as in Example 13.

Example 15. Emulsion In addition to using 2.0 parts of L-ascorbic acid-2-phosphate and 2.0 parts of sodium L-ascorbic acid-2-phosphate instead of 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 parts of potassium hydroxide in the component B of Example 13 Obtained an emulsion in the same manner as in Example 5.

Example 16 Emulsion In the same manner as in Example 13, except that 2.0 parts of arbutin is used instead of 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 parts of potassium hydroxide in the component B of Example 13. Obtained.

Example 17. Latex In the component B of Example 13, in place of 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 parts of potassium hydroxide, a rice bran extract hydrolyzate (trade name “Grace Snow” manufactured by Technoble Co., Ltd.) An emulsion was obtained in the same manner as in Example 13 except that 5.0 parts of * snow * HP ", solid content concentration 3.5%) were used.

Example 18 Emulsion White component extract (trade name “Sinablanca-WH” manufactured by Technoble Co., Ltd.) instead of 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 part of potassium hydroxide in the B component of Example 13 The emulsion was obtained in the same manner as in Example 13 except that 5.0 parts of a solid content concentration of 1.0% was used.

Example 19. Emulsion Example 13 except for using 1.0 part of γ-amino-β-hydroxybutyric acid in place of 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 part of potassium hydroxide in the component B of Example 13 An emulsion was obtained in the same manner as in Example 13.

Example 20. Emulsion Emulsion in the same manner as in Example 13 except that 2.0 parts of L-ascorbic acid-2-glucoside and 0.5 parts of potassium hydroxide are used in the component B of Example 5 instead of 2.0 parts of potassium hydroxide. Got.

Example 21. 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] Part Enzyme hydrolyzate of Production Example 1 5.0
Sodium carboxymethylcellulose 0.2
Bentonite 0.5
Propylene glycol 4.0
Triethanolamine 1.1
Methylparaben 0.1
Purified water Amount that makes the total amount 100 parts [Component C] Part Titanium oxide 8.0
Talc 4.0
Coloring pigment appropriate amount The 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.

Example 22. Cream foundation [component A] part Stearic acid 5.0
Cetanol 2.0
Glyceryl monostearate 3.0
Liquid paraffin 5.0
Squalane 3.0
Isopropyl myristate 8.0
Polyoxyethylene (20) glyceryl monostearate 2.0
Propylparaben 0.1
[Component B] Part Enzyme hydrolyzate of Production Example 1 5.0
Sorbitol 3.0
1,3-butylene glycol 5.0
Triethanolamine 1.5
Methylparaben 0.1
Purified water Amount that makes the total amount 100 parts [Component C] Part Titanium oxide 8.0
Talc 2.0
Kaolin 5.0
Bentonite 1.0
Coloring pigment appropriate amount [Component D]
Fragrance 0.3
Component C was mixed and pulverized with a pulverizer. The component B was mixed, and the pulverized component C was added thereto and uniformly dispersed in a colloid mill. The components A and B and C dispersed uniformly were each heated to 80 ° C., and then the components A were added to the components B and C while stirring, and further homogenized with Hiscotron (5000 rpm) for 2 minutes. After cooling this to 50 degreeC, D component was added and stirred and mixed, and also it cooled to 30 degrees C or less, stirring, and obtained the cream foundation.

Example 23. 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 Enzyme hydrolyzate of Production Example 1 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 Skin irritation test (SIRC irritation evaluation test)
Using the enzyme hydrolyzate of the salamander seeds of Production Example 1 and the extract of the salamander seeds of Comparative Production Example 1 as samples, their skin irritation was examined.
[Test method]

Rabbit eye mucosal cells SIRC were seeded at 5 × 10 ³ cells / well in a 96-well microplate containing Eagle's minimum essential medium containing 10% FBS, and pre-cultured at 37 ° C. and 5.0% CO ₂ for 3 days. Then, a sample solution prepared by adding a 1,3-butylene glycol solution to the enzyme hydrolyzate of Production Example 1 and the extract of Comparative Production Example 1 (the concentration of 1,3-butylene glycol solution is 30%) Were added to the medium so that the concentrations (as solutions) were 2.5%, 5.0%, and 10.0%, respectively, and the cells were further cultured under the same conditions for 2 days. Here, the concentration of the solution when added to the medium is the concentration of the enzyme hydrolyzate of Production Example 1 and the extract of Comparative Production Example 1 as a sample solution, and each solid content concentration is a production example. In the enzyme hydrolyzate of No. 1, 0.017% when the solution concentration is 2.5%, 0.034% when the solution concentration is 5.0%, and 0 when the solution concentration is 10.0%. 0.068% in the extract of Comparative Production Example 1 when the solution concentration is 2.5%, 0.032% when the solution concentration is 5.0%, and the solution concentration is 10%. In the case of 0.0%, it becomes 0.064%. After culturing for 2 days, the medium was removed, 200 μL of a 0.0052% neutral red-containing medium was added and maintained at 37 ° C. for 3 hours, and then the neutral red incorporated into the cells was extracted with 1% acetic acid-containing ethanol. , Microplate reader (Model
680, manufactured by Bio-Rad), and the amount of neutral red uptake was measured at a wavelength of 540 nm. Further, instead of the sample solution, as a control, the same operation as described above was performed when a 30% 1,3-butylene glycol solution was added in an amount equal to each sample concentration, and this was used as a control. Here, since neutral red is a soluble dye taken up by cells that have not been damaged, it is possible to test the stimulation of each sample on cells by the amount of the dye taken up. As a measurement result, the relative value of each sample solution when the control was 100% was defined as the neutral red uptake rate (%).

[result]
The results of Test Example 1 are shown in Table 1.
[Table 1]

As shown in Table 1, when the extract of the salamander seed of Comparative Production Example 1 was used as a sample, the uptake rate of neutral red was significantly reduced in a concentration-dependent manner. It became clear that it was expensive. On the other hand, since a large amount of neutral red is incorporated in the enzyme hydrolyzate of the seed of the salamander of Production Example 1 according to the present invention as compared with the above extract that is a non-enzymatic treatment product, It was revealed that the skin irritation was remarkably suppressed by subjecting the salamander seed extract to enzymatic hydrolysis.

Test Example 2 Measurement of Ceramide Synthase (β-Glucocerebrosidase) Activity Using the enzyme hydrolyzate of the salamander seeds of Production Example 1 and the extract of the salamander seeds of Comparative Production Example 1 as samples, their β-glucocerebros Sidase activity promoting effect was examined.
[Test method]
Human epidermal cells PHK16-0b were seeded at 1 × 10 ⁴ cells / hole in a 96-well microplate. As the medium, MCDB153 medium (manufactured by SIGMA) plus additive epidercel HKGS (manufactured by Kurabo Industries) was used. This was pre-cultured under conditions of 37 ° C. and 5.0% CO ₂ for 3 days, and then the enzyme hydrolyzate of Production Example 1 and the extract of Comparative Production Example 1 were used as samples. Concentration (the concentration is the concentration of the enzyme hydrolyzate of Production Example 1 and the extract of Comparative Production Example 1 as a solution, and the solid concentration of each is the concentration of the solution in the enzyme hydrolyzate of Production Example 1 Is 0.017% when the concentration is 2.5%, and 0.016% when the solution concentration is 2.5% in the extract of Comparative Production Example 1). The cells were further cultured for 4 days under the same conditions. Next, after removing the medium and washing with PBS (−), a PMSF-containing 1% Triton X-100 solution was added, and the cells were subjected to a lysis operation to obtain an enzyme solution. Next, a substrate solution (1 mM 4-Methylumbelliferyl-β-Glucopyranoside) was added, and the mixture was reacted at 37 ° C. for 1 hour while gently stirring with a vortex mixer. After stopping the reaction by adding a reaction stop solution (0.2 M carbonate bicarbonate buffer (pH 10.5)), fluorescence intensity (excitation: 355 nm, emission: 460 nm: fluorescence microplate reader (Fluoroscan Ascent, manufactured by Thermo Fisher Scientific) ) Was measured. In addition, in the case of no sample added with PBS (-) instead of the sample solution (control), the same operation as described above was performed, and the relative value of the fluorescence intensity at the time of adding each sample to the obtained fluorescence intensity. The β-glucocerebrosidase activity enhancement rate (%) in epidermal cells was obtained. Moreover, in order to confirm whether the test system is functioning normally, the same test was performed when 0.0025% β-galactocerebroside was added as a positive control instead of the sample solution.

[result]
The results are shown in Table 2.
[Table 2]

  As shown in Table 2, in the enzyme hydrolyzate of the salamander seed of Production Example 1, β-glucocerebrosidase activity promotion that the extract of the salamander seed of Comparative Production Example 1 did not have It became clear that a new action was created. In addition, when β-galactocerebroside, which is a positive control, was used as a sample, β-glucocerebrosidase activity was also confirmed, and it was confirmed that this test system was performed normally.

Test Example 3 Epidermal cell activation test Using the enzyme hydrolyzate of the salamander seeds of Production Example 1 and the extract of the salamander seeds of Comparative Example 1 as samples, their epidermal cell activation effects were examined.
[Test method]
Human epidermal cells PHK16-0b (Lot.040817 (4)) were seeded in a 96-well microplate at a concentration of 1 × 10 ⁴ cells / well. As the medium, MCDB153 (manufactured by SIGMA) to which Epidercel HKGS (manufactured by Kurabo Industries) was added as a growth promoter was used. After pre-culturing at 37 ° C. for 2 days, the enzyme hydrolyzate of Production Example 1 and the extract of Comparative Production Example 1 were used as samples, and the samples were 2.5% and 5.0% in concentration (the concentrations are the production examples) The concentration of each of the enzyme hydrolyzate of 1 and the extract of Comparative Production Example 1 as a solution, and the solid content concentration of each of the enzyme hydrolyzate of Production Example 1 is 2.5%. 0.017%, 0.034% when the solution concentration is 5.0%, and 0.016% when the solution concentration is 2.5% in the extract of Comparative Production Example 1. In the case of 5.0%, it was 0.032%), and the mixture was further cultured at 37 ° C. for 4 days. Next, the medium was removed, and a 0.03% MTT solution prepared using PBS (−) was added and maintained at 37 ° C. Then, using a microplate reader (MODEL 680, manufactured by Bio-Rad), a wavelength of 570 The MTT value was measured at −630 nm. In the case of no sample addition (control), the same operation as described above was performed, and the relative value of the MTT value at the time of each sample addition with respect to the MTT value obtained here was determined to be the human epidermal cell MTT activity rate (%). . For comparison, the same test was performed when 100 mM glucose was added instead of the sample solution (positive control).

[result]
The results are shown in Table 3.
[Table 3]

  As shown in Table 3, the enzyme hydrolyzate of the salamander seeds of Production Example 1 was significantly superior in cover cell activation compared to the extract of the salamander seeds of Comparative Production Example 1 in a concentration-dependent manner. It became clear to show the effect. In addition, also when using glucose which is a positive control as a sample, since the result which shows epidermal cell activation was confirmed, it was also confirmed that this test system was performed normally.

In addition, it was also clarified that the enzyme hydrolyzate of the salamander seeds of Production Example 1 significantly suppresses the coloration and starch that occur over time in the extract of the salamander seeds of Comparative Production Example 1.

Claims (2)

Cosmetics containing an enzyme hydrolyzate obtained by hydrolyzing the extract of Zanthoxylum piperitum, a plant of the genus Rutaceae, Zanthoxylum, with proteolytic enzymes and lipolytic enzymes Fee. The cosmetic according to claim 1, wherein the enzyme hydrolyzate is obtained by hydrolyzing a salamander seed extract with an enzyme.