JP7550430B2 - Protease activity enhancer - Google Patents

Description

The present invention relates to a protease activity enhancer that is highly effective and safe and can be incorporated into preparations such as topical skin preparations.

In recent years, there has been a demand for functional materials that prevent and improve aging at the cellular level by improving and activating cell function, or by expelling or removing substances that cause cellular aging (such as active oxygen), but no such materials have been found that fully meet the requirements for stability, safety, and effectiveness.

In order to solve the above problems, we discovered that an enzymatic hydrolysate of rice, a natural product, has the effect of preventing and improving cell aging by enhancing protease activity, which led to the present invention.

Although Patent Documents 1 to 3 disclose that rice-derived components prevent and improve aging of skin and hair, it was not known that enzymatic hydrolysates of rice or rice extracts have the effect of enhancing protease activity.
JP 04-117318 A JP 05-221844 A Patent Publication No. 2013-224288

The present invention is a protease activity enhancer containing, as an active ingredient, an enzymatic hydrolysate of rice or a rice extract.
In the present invention, the rice is preferably pigmented rice.
In the present invention, the hydrolysate is preferably obtained by the use of two or more proteases.

The protease activity enhancer of the present invention can prevent and improve cellular aging because the active ingredient, rice hydrolysate, enhances intracellular protease activity.

The present invention is a protease activity enhancer that contains as an active ingredient a hydrolysate obtained by hydrolyzing a rice extract.

The "rice" used in the present invention refers to the seeds of the rice plant "Oryza sativa linne (Gramineae)" and includes white rice, as well as black rice (purple black rice), purple rice, red rice, and green rice. The rice used in the extract of the present invention may be either brown rice or polished rice.

In the present invention, examples of the solvents used in the extraction process include purified water; lower alcohols such as ethanol; polyols such as ethylene glycol, propylene glycol, glycerin, and 1,3-butylene glycol; higher alcohols such as oleyl alcohol, stearyl alcohol, and octyldodecanol; ketones such as acetone; esters such as ethyl acetate; and hydrocarbon solvents such as hexane, chloroform, and benzene, which may be used alone or in combination of two or more. Of these, in terms of their wide application to cosmetics, purified water or a mixture of purified water with one or more of ethanol, glycerin, and 1,3-butylene glycol is preferred.

When using these mixed solvents, the volume ratio is preferably 1 to 25:1 for purified water and ethanol, 1 to 15:1 for purified water and glycerin, and 1 to 15:1 for purified water and 1,3-butylene glycol.

The pH may be adjusted to be acidic, neutral, or alkaline, but it is preferable to adjust it to be alkaline from the viewpoint of the effect to be obtained and ease of use as an agent for preventing and improving color changes. Examples of alkaline adjusters include sodium salts such as sodium hydroxide and sodium carbonate, and potassium salts such as potassium hydroxide, and the pH of these is set to 7.5 to 14.0. Of these, sodium hydroxide and sodium carbonate are preferred because they can be set to the desired pH at low concentrations.

The extraction conditions, such as extraction temperature and extraction time, vary depending on the type and pH of the solvent used. For example, when using purified water as the extraction solvent and extracting at a pH of 7.5 to 14.0, the extraction temperature is preferably 4 to 40°C, more preferably 15 to 25°C. The extraction time is preferably 4 hours to 7 days, more preferably 20 to 24 hours.

Next, the extract obtained as described above is subjected to hydrolysis treatment. Examples of the treatment method include acid treatment, alkali treatment, enzyme treatment, etc., among which enzyme treatment is the most preferable. The enzyme to be used may be one or more selected from proteolytic enzymes, amylolytic enzymes, cellulose-degrading enzymes, and lipolytic enzymes.

The proteolytic enzyme may be any of enzymes derived from animals, plants, and microorganisms. Examples include actinases, pepsins such as pepsin, trypsins such as trypsin, papains such as papain and chymopapain, peptidases such as glycylglycine peptidase, carboxypeptidase, aminopeptidase, and bromelain, and one or more of these may be used. Examples of starch-degrading enzymes include glucoamylase and α-amylase. Examples of cellulose-degrading enzymes include cellulase, hemicellulase, pectinase, and the like. Examples of fat-degrading enzymes include lipase. As the enzymes used in the present invention, it is preferable to use one or a combination of two or more proteolytic enzymes.

The amount of the proteolytic enzyme used is 0.0005 to 0.05 parts, preferably 0.001 to 0.005 parts, per 100 parts of extract (parts by weight, the same applies below) per treatment. When using two or more types of enzymes, a total amount of about 0.003 to 0.015 parts is preferred in terms of effectiveness.

The enzyme treatment time varies depending on the type of protease used and the decomposition temperature, but is usually preferably 1 to 24 hours. The decomposition temperature is about 30 to 50°C. The decomposition product thus obtained may be concentrated under reduced pressure to adjust the concentration, or may be powdered by freeze-drying or spray-drying for use.

It is preferable that the pH of the resulting decomposition product solution be adjusted to pH 4 to 8 to ensure safety to the skin.

When the protease activity enhancer of the present invention is incorporated into a preparation for topical skin applications (drugs, quasi-drugs, or cosmetics) or an oral cosmetic preparation, ingredients that are typically used in preparations for topical skin applications, such as oily ingredients, surfactants (synthetic and natural), moisturizers, anti-inflammatory agents, thickeners, preservatives/bactericides, anti-acne agents, cell activators, powder ingredients, UV absorbers, antioxidants, whitening agents, anti-wrinkle agents, pigments, fragrances, etc., can be appropriately incorporated as necessary.

Here, examples of oily components include olive oil, jojoba oil, castor oil, soybean oil, rice oil, rice germ oil, coconut oil, palm oil, cocoa oil, meadowfoam oil, shea butter, tea tree oil, avocado oil, macadamia nut oil, bergamot oil, lavender oil, rose oil, bergamot oil, chamomile oil, and other plant-derived oils and fats such as squalane; vitamin A oil; animal-derived oils and fats such as mink oil and turtle oil; waxes such as beeswax, carnauba wax, rice wax, and lanolin; liquid paraffin, petrolatum, and paraffin wax. Examples of such compounds include hydrocarbons such as cocos and squalane; fatty acids such as myristic acid, palmitic acid, stearic acid, oleic acid, isostearic acid, and cis-11-eicosenoic acid; higher alcohols such as lauryl alcohol, cetanol, pantothenyl alcohol, and stearyl alcohol; synthetic esters and synthetic triglycerides such as isopropyl myristate, isopropyl palmitate, butyl oleate, 2-ethylhexyl glyceride, and higher fatty acid octyldodecyl (e.g., octyldodecyl stearate).

Examples of surfactants include nonionic surfactants such as polyoxyethylene alkyl ethers, polyoxyethylene fatty acid esters, polyoxyethylene sorbitan fatty acid esters, glycerin fatty acid esters, polyglycerin fatty acid esters, polyoxyethylene glycerin fatty acid esters, polyoxyethylene hydrogenated castor oil, and polyoxyethylene 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 Anionic surfactants such as phenyl alkyl phenyl ether phosphate; cationic surfactants such as quaternary ammonium salts, primary to tertiary fatty amine salts, trialkyl benzyl ammonium salts, alkyl pyridinium salts, 2-alkyl-1-alkyl-1-hydroxyethyl imidazolinium salts, N,N-dialkyl morpholinium salts, and polyethylene polyamine fatty acid amide salts; amphoteric surfactants such as N,N-dimethyl-N-alkyl-N-carboxymethyl ammonio betaine, N,N,N-trialkyl-N-alkylene ammonio carboxybetaine, N-acylamidopropyl-N', N'-dimethyl-N'-β-hydroxypropyl ammonio sulfobetaine, etc. can be used.

As emulsifiers and/or emulsifier aids, stevia derivatives such as enzyme-treated stevia, saponin or its derivatives, casein or its salts (sodium, etc.), sugar and protein complexes, sucrose or its esters, lactose, soy-derived water-soluble polysaccharides, soy-derived protein and polysaccharide complexes, lanolin or its derivatives, cholesterol, stevia derivatives (enzyme-treated stevia, etc.), silicates (aluminum, magnesium, etc.), carbonates (calcium, sodium, etc.), saponin and its derivatives, lecithin and its derivatives (hydrogenated lecithin, etc.), lactic acid bacteria fermented rice, lactic acid bacteria fermented germinated rice, lactic acid bacteria fermented grains (wheat, beans, millet, etc.), etc. can also be blended.

Examples of moisturizing agents include glycerin, propylene glycol, dipropylene glycol, 1,3-butylene glycol, polyethylene glycol, sorbitol, xylitol, sodium pyrrolidone carboxylate, and the like, as well as sugars such as trehalose and raffinose, mucopolysaccharides (e.g., hyaluronic acid and its derivatives, hyaluronic acid fermentation liquid, chondroitin and its derivatives, heparin and its derivatives, etc.), elastin and its derivatives, collagen and its derivatives, collagen peptides, NMF-related substances, lactic acid, urea, higher fatty acid octyldodecyl, seaweed extract, estradiol, various amino acids and their derivatives.

Examples of thickeners include components derived from brown algae, green algae, or red algae, such as alginic acid, agar, carrageenan, and fucoidan; polysaccharides such as pectin and aloe polysaccharides; gums such as tragacanth gum, locust bean gum, xanthan gum, and guar gum; cellulose derivatives such as carboxymethylcellulose, hydroxyethylcellulose, and hydroxypropylcellulose; synthetic polymers such as carboxyvinyl polymers, alkyl-modified carboxyvinyl polymers, polyvinyl alcohol, polyvinylpyrrolidone, and acrylic acid/methacrylic acid copolymers; hyaluronic acid and its derivatives; polyglutamic acid and its derivatives, and polyacrylic acid.

Anti-inflammatory agents include allantoin, dipotassium glycyrrhizinate, monoammonium glycyrrhizinate, β-glycyrrhetinic acid, stearyl glycyrrhetinate, ε-aminocaproic acid, d-camphor, dl-camphor, zinc oxide, panthenol, pyridoxine hydrochloride, and riboflavin or its derivatives.

Examples of preservatives and disinfectants include urea; benzoic acid or its salts, paraoxybenzoic acid esters such as methyl paraoxybenzoate, ethyl paraoxybenzoate, propyl paraoxybenzoate, and butyl paraoxybenzoate; phenoxyethanol, dichlorophene, hexachlorophene, chlorhexidine hydrochloride, benzalkonium chloride, salicylic acid, sodium salicylate, zinc pyrithione, benzalkonium chloride, ethanol, undecylenic acid, phenols, and alkyl bromides. These include isoquinolinium, resorcinol, jamal (imidazolidinyl urea), isopropyl methylphenol, triclosan, trichlorocarbanide, trichlorohydroxydiphenol ether, hinokitiol, 1,2-pentanediol, propanediol, concentrated benzalkonium chloride solution 50, essential oils such as peppermint oil and eucalyptus oil, bark distillate, fermented radish liquid, ethanol derived from plants such as sugar cane and corn, or 1,3-butylene glycol.

Cell activators include pantothenyl alcohol, menthol, dl-menthol, and gamma-oryzanol.

Anti-acne agents include sulfur, salicylic acid or its salts, photosensitizer No. 201, pyridoxine dicaprylate, etc.

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, cellulose-based powder, grain powder (rice, wheat, corn, millet, etc.), and bean powder (soybean, adzuki bean, etc.).

Examples of ultraviolet absorbers include ethyl paraaminobenzoate, ethylhexyl paradimethylaminobenzoate, amyl salicylate and its derivatives, 2-ethylhexyl paramethoxycinnamate, octyl cinnamate, oxybenzone, 2,4-dihydroxybenzophenone, 2-hydroxy-4-methoxybenzophenone-5-sulfonate, 4-tert-butyl-4-methoxybenzoylmethane, 2-(2-hydroxy-5-methylphenyl)benzotriazole, urocanic acid, ethyl urocanate, aloe extract, etc.

Examples of antioxidants include butyl hydroxyanisole, butyl hydroxytoluene, propyl gallate, carotenoids such as astaxanthin, vitamin E and its derivatives (e.g., tocopherol acetate, tocopherol nicotinate), vitamin A and its derivatives (retinol palmitate, etc.), etc.

Also, examples of whitening agents include one or more selected from ellagic acid and its derivatives, resorcinol derivatives, potassium 4-methoxysalicylic acid, magnolignan (5,5'-dipropyl-biphenyl-2,2'-diol), hydroxybenzoic acid and its derivatives, vitamin E and its derivatives, alpha-hydroxy acid, and AMP (adenosine monophosphate, adenosine monophosphate).

Examples of resorcinol derivatives include 4-n-butylresorcinol and 4-isoamylresorcinol. Examples of 2,5-dihydroxybenzoic acid derivatives include 2,5-diacetoxybenzoic acid, 2-acetoxy-5-hydroxybenzoic acid and 2-hydroxy-5-propionyloxybenzoic acid. Examples of α-hydroxy acids include lactic acid, malic acid, succinic acid, citric acid and α-hydroxyoctanoic acid.

In addition, the following animal, plant or microorganism-derived ingredients can also be used in combination. For example, collagen or its hydrolysate, yeast extract or hydrolysate, lactic acid bacteria culture, Gramineae plants, Cruciferae plants, Theaceae plants, Rosaceae plants, Paeoniaceae plants, Rutaceae plants, Amaranthaceae plants, Leguminosae plants, Asteraceae plants, Fabaceae plants, Malvaceae plants, Gentianaceae plants, Lamiaceae plants, Nelumbaceae plants, Cucurbitaceae plants, Araliaceae plants, Solanaceae plants, Bignoniaceae plants, Actinidiaceae plants, Mulberry plants, Iridaceae plants, Campanulaceae plants, Oleaceae plants, Actinidiaceae plants, Mulberry plants, Rhamnaceae plants, Orchidaceae plants, Anacardiaceae plants extracts or hydrolysates or fermented products of one or more plants selected from the family Gardenia, Valencaceae, Rutaceae, Myrtaceae, Liliaceae, Crassulaceae, Cupressaceae, Convolvulaceae, and Asparagaceae; extracts or hydrolysates or fermented products of one or more seaweeds selected from the family Laminaria, Laminaceae, and Ulvulaceae; jellyfish (autolyzed products of moon jellyfish, Nomura's jellyfish, etc.); hydrolysates or fermented products of hyaluronic acid; and extracts or hydrolysates or fermented products of royal jelly.

As the components derived from plants of the Gramineae family, in particular, hydrolyzed rice leaves, hydrolyzed rice bran extract, hydrolyzed germinated brown rice, fermented rice liquid, sake lees extract derived from sake, extract of bamboo shoot skin of Madake or Moso bamboo, and fermented pearl barley seeds are preferred. As the components derived from plants of the Brassicaceae family, in particular, extracts of seeds of Hakugai, Ogai or Kokugai or their hydrolyzates or fermented products are preferred. As the components derived from plants of the Theaceae family, in particular, green tea (Yabukita, Samidori, Asahi, Goko, Ujimidori, Kyomidori, Ujihikari, Samidori, Benifuki, etc.) and black tea (Darjeeling, Assam, Ceylon, Earl Grey, Honey-scented black tea, etc.) are preferred. As the components derived from plants of the Rosaceae family, extracts of Damask rose flowers, extracts of peach flowers, leaves or immature fruits, extracts of apricot fruits or seeds, extracts of strawberry flowers, and extracts of cherry blossoms or leaves are preferred. In addition, as the component derived from a plant of the family Paeoniaceae, an extract of the root or flower of the peony and an extract of the flower or root of the peony are preferable. In addition, as the component derived from a plant of the family Amaranthaceae, an extract of Salicornia herbacea is particularly preferable. In addition, as the component derived from a plant of the family Zosteraea, an extract of Zostera marina or Zostera koa is particularly preferable. In addition, as the component derived from a plant of the family Fabaceae, an extract of white soybean or black soybean or a hydrolyzate thereof, or a fermented soy milk liquid, an extract of adzuki bean, an extract of red clover, and an extract of pueraria lobata root are particularly preferable. In addition, as the component derived from a plant of the family Asteraceae, an extract of burdock root, an extract of sunflower sprouts, an extract of Lamium arbutifolia, an extract of Arnica officinalis, or an extract of chamomile flower are particularly preferable. In addition, as the component derived from a plant of the family Malvaceae, a fermented product of hibiscus, Rose of Sharon, or Hibiscus is preferable. In addition, as the component derived from a plant of the family Gentianaceae, an extract of Gentiana serrata is preferable. In addition, as the plant of the family Lamiaceae, an extract of Perilla frutescens and an extract of the fruit of the family Caryophyllus are preferable. As the component derived from a plant of the family Nelumboceae, particularly, lotus flower or lotus seed extract or lotus seed fermentation product is preferable. As the component derived from a plant of the family Cucurbitaceae, particularly, loofah extract is preferable. As the component derived from a plant of the family Araliaceae, an extract or fermentation product of Panax ginseng is preferable. As the component derived from a plant of the family Solanaceae, an extract of eggplant (long eggplant, water eggplant, rice eggplant, Kamo eggplant, etc.) is preferable. As the component derived from a plant of the family Bignoniaceae, a bark extract of Pau d'Arco is preferable. As the component derived from a plant of the family Actinidiaceae, an extract of immature kiwi is preferable. As the component derived from a plant of the family Mulberry, a mulberry fruit extract, an extract of the fruit or bark of fig is preferable. As the component derived from a plant of the family Rhamnaceae, a jujube fruit extract is preferable. As the component derived from a plant of the family Iridaceae, a saffron is preferable. As the component derived from a plant of the family Campanulaceae, an extract or hydrolysate of the root of Lycoris nigricans is preferable. As the component derived from a plant of the family Anacardiaceae, a mango fruit extract is preferable. As the component derived from a plant of the family Garcinia, mangosteen fruit extract is particularly preferred. As the component derived from a plant of the family Valenciaceae, cherimoya fruit extract is particularly preferred. As the component derived from a plant of the family Rutaceae, Satsuma mandarin, bergamot fruit extract, grapefruit or banpeiyu fruit (including immature fruit), or Zanthoxylum pepper seed extract is particularly preferred. As the component derived from a plant of the family Liliaceae, an extract of daylily, daylily, Casablanca, Madonna lily, or lily of the valley is particularly preferred. As the component derived from a plant of the family Crassulaceae, an extract or fermented product of Rhodiola rosea (Rhodiola rosea) is particularly preferred. As the component derived from a plant of the family Oleaceae, jasmine flower extract is particularly preferred. As the plant of the family Cupressaceae, Juniperus communis fruit extract is particularly preferred. As the component derived from a plant of the family Myrtaceae, guava leaf extract is particularly preferred. As the plant of the family Orchidaceae, an extract of the root (white and white) of Bletilla serrata is particularly preferred. As the component derived from a plant of the Convolvulaceae family, a sweet potato extract or its fermentation product, or a sweet potato shochu lees extract or its fermentation product is preferable. As the plant of the Asparagaceae family, asparagus (green asparagus and white asparagus) is preferable. As the component derived from seaweed of the Laminaceae family, a kelp extract is particularly preferable, as the component derived from seaweed of the Mirinaceae family, a katamen kirinsai extract is particularly preferable, and as the component derived from seaweed of the Ulva family, an Ulva pertusa extract is particularly preferable. As the component derived from seaweed of the Funoriaceae family, a funori extract is particularly preferable.

The present invention will now be described in more detail with reference to manufacturing examples, formulation examples, and test examples, but the present invention is not limited thereto. In the following, all parts are by weight, and all percentages are by weight.

The present invention will now be described in more detail with reference to manufacturing examples, working examples (formulation examples) and test examples, but the present invention is not limited thereto. In the following, all parts are by weight and all percentages are by weight.

Production Example 1. Preparation of rice hydrolysate solution (1)
1000g of 0.1% sodium hydroxide solution was added to 250g of polished black rice, and the mixture was stirred and extracted for one day, after which the remaining black rice residue was removed by rough filtration through a filter cloth. The extract was neutralized with dilute hydrochloric acid, and proteolytic enzymes (actinase AS 0.02%, papain 0.02%) were added based on the amount of the liquid, and the enzyme was hydrolyzed at 40°C for 2 hours, then heated at 80°C for 1 hour to inactivate the enzyme, and cooled to room temperature. The enzyme-treated liquid thus obtained was purified and filtered to obtain 805g of a light brown transparent black rice hydrolysate solution (solid content: 1.70%).

Production Example 2. Preparation of rice hydrolysate solution (2)
The same procedure as in Production Example 1 was repeated except that bromelain was used instead of papain, to obtain 797 g of a light brown, transparent black rice hydrolysate solution (solids concentration: 1.68%).

Production Example 3. Preparation of rice hydrolysate solution (3)
The same procedure as in Production Example 1 was repeated except that red rice was used instead of the black rice used in Production Example 1, to obtain 785 g of a pale red, transparent red rice hydrolysate solution (solid content: 1.59%).

Production Example 4. Preparation of white rice hydrolysate solution (4)
The same procedure as in Production Example 1 was repeated except that white rice was used instead of the black rice used in Production Example 1, to obtain 790 g of a pale yellow, transparent rice hydrolysate solution (solid content: 1.65%).

Test Example 1. Protease activity enhancement test Human dermis-derived fibroblasts NB1RGB were seeded at 1 x ¹⁰⁴ cells/well in a 96-well microplate containing 0.5% NCS-containing Eagle's minimum essential medium, and pre-cultured for 1 day under conditions of 37°C and 5.0% _CO2 . The hydrolysate solutions of Production Examples 1 to 3 were then added to the medium at a concentration of 2.0% (final concentration as a solution relative to the total amount of medium), and cultured under the same conditions for another 3 days. After that, the cell culture supernatants of each test group were removed, and the cells were disrupted using 100μL of cell lysis solution (Tris-HCL buffer (pH 8.0) containing 1% Tween 20: 12mM trishydroxymethylaminomethane, 5mM EDTA, 150mM NaCl, hydrochloric acid) to prepare a crude enzyme solution. 50μL of the crude enzyme solution was transferred to a separate plate, and 10μL of a buffer containing 65μM Suc-Leu-Leu-Val-Tyr-AMC as a substrate was added thereto, and the mixture was allowed to react at 37℃ for 1 hour. After the reaction, the fluorescence intensity (excitation wavelength: 355nm, fluorescence wavelength: 460nm) was measured using a fluorescent plate reader (Fluoroscan Ascent, Thermo Labsystems) and this was taken as the protease. The same procedure as above was also carried out for the control case where PBS(-) was added instead of the sample solution, and the relative value of the protease activity when each sample was added to the protease activity obtained here was calculated, and this was taken as the protease activity rate (%).

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

As shown in Table 1, it was confirmed that the hydrolysate according to the present invention has the effect of enhancing intracellular protease activity. This suggests that it exerts an anti-aging effect by activating the protein degradation system within the cells, promoting metabolism or the removal of waste products, and maintaining the health of the cells.

Formulation Example 1. Lotion [Ingredients] Parts Eucalyptus oil 0.2
Polyoxyethylene (5.5) Cetyl Alcohol 5.0
Hydrolyzate of Production Example 1 0.1
Tocopherol acetate 0.02
Dipotassium glycyrrhizinate 0.5
Monoammonium glycyrrhizinate 0.5
Stearyl glycyrrhizinate 0.05
Isopropyl methylphenol 0.1
Aligned in 0.1
D-pantothenyl alcohol 0.1
Salicylic acid 0.5
Urea 5.0
l-Menthol 0.9
dl-Menthol 0.2
1,3-butylene glycol 5.0
Sodium citrate 0.2
Methylparaben 0.1
Hinokitiol 0.003
Photosensor No. 201 0.002
Purified water (enough to make the total amount 100 parts)

Formulation Example 2. Lotion A lotion of Formulation Example 2 was obtained in the same manner as in Formulation Example 1, except that the hydrolysate of Production Example 2 was used in place of the hydrolysate of Production Example 1 among the ingredients of Formulation Example 1.

Formulation Example 3. Lotion A lotion of Formulation Example 3 was obtained in the same manner as in Formulation Example 1, except that the hydrolysate of Production Example 3 was used in place of the hydrolysate of Production Example 1 among the ingredients of Formulation Example 1.

Formulation Example 4. Lotion [Ingredients] Parts Glyceryl caprylate 3.0
Polyglyceryl-10 Laurate 3.0
Cetanol 2.0
Behenyl alcohol 2.0
Methylparaben 0.1
Hydrolyzate of Production Example 4 0.1
Ascorbic Acid 3.0
Glycyrrhizic acid 0.5
β-Glycyrrhetinic acid 0.05
Tocopherol nicotinate 0.1
Resorcinol 0.1
Zinc oxide 2.0
dl-Camphor 0.5
Glycerin 2.0
1,3-butylene glycol 5.0
Potassium hydroxide 0.5
Purified water (enough to make the total amount 100 parts)

Formulation Example 5. Lotion [Ingredients] Parts Jojoba oil 1.0
Polyoxyethylene (5.5) Cetyl Alcohol 5.0
Methylparaben 0.1
Hydrolyzate according to Production Example 2 0.2
Ascorbic acid glucoside 2.0
Tranexamic acid 2.0
ε-aminocaproic acid 0.1
Sulfur 0.2
Estradiol 0.1
Glycerin 5.0
1,3-butylene glycol 5.0
Sodium citrate 0.2
Sodium metabisulfite 0.2
d-Camphor 0.1
Purified water (enough to make the total amount 100 parts)

Formulation Example 6. Emulsion [Ingredients] Parts Squalane 5.0
Cyclopentane siloxane 1.0
Hexalan 3.0
Hexyldecyl isostearate 1.0
Caprylic/capric triglyceride 1.0
Polyglyceryl-10 Laurate 5.0
Polyglyceryl-10 Isostearate 5.0
Ascorbyl dipalmitate 15.0
Hydrogenated soy lecithin 1.5
Hydrolysate of Production Example 1 2.0
Ascorbic acid phosphate magnesium salt 3.0
Arbutin 3.0
Potassium hydroxide 0.5
Glycerin 3.0
1,3-butylene glycol 2.0
Carboxymethyl cellulose 0.3
Xanthan gum 0.2
Tremella polysaccharide 0.2
Sodium hyaluronate 0.01
Tocopherol acetate 0.3
Tocopherol nicotinate 0.1
Glycyrrhizic acid 0.1
Dipotassium glycyrrhizinate 0.1
Isopropyl methylphenol 0.1
Water-soluble collagen 1.0
Hydrolyzed Collagen 1.0
Sodium hyaluronate 1.0
Purified water (enough to make the total amount 100 parts)

Formulation Example 7. Milky Lotion A milky lotion was obtained in the same manner as in Formulation Example 6, except that 2.0 parts of L-ascorbic acid-2-glucoside was used in place of 2.0 parts of magnesium ascorbyl phosphate.

Formulation Example 8. Milky Lotion A milky lotion was obtained in the same manner as in Formulation Example 6, except that 2.0 parts of tranexamic acid were used instead of 2.0 parts of magnesium ascorbyl phosphate and 0.5 parts of potassium hydroxide.

Formulation Example 9: Milky Lotion A milky lotion was obtained in the same manner as in Formulation Example 6, except that 3.0 parts of niacinamide was used in place of 2.0 parts of magnesium ascorbyl phosphate among the ingredients in Formulation Example 6.

Formulation Example 10. Cream [Ingredients] Parts Olive oil 5.0
Jojoba oil 5.0
Squalane 5.0
Hexyldecyl isostearate 5.0
Lauroyl glutamate di(octyldodecyl/phytosteryl/behenyl) 5.0
Glyceryl Caprylate 1.0
Glyceryl stearate 1.0
Isostearyl glyceryl 3.0
γ-oryzanol 0.1
Behenyl alcohol 2.0
Palmitic acid 2.5
D-pantothenyl alcohol 3.0
Allantoin 0.1
Riboflavin 0.01
Resorcinol 0.1
Benzalkonium chloride 0.05
Urea 3.0
β-Glycyrrhetinic acid 0.1
Stearyl glycyrrhetinate 0.1
Ammonium glycyrrhizinate 0.1
Hydrolysate of Production Example 1 3.0
Lactic acid fermented rice 2.0
Hydrogenated lecithin 0.5
Hydrogenated lysolecithin 0.5
Oil-soluble ginseng extract 2.0
Xanthan gum 1.0
Zinc oxide 0.5
dl-Camphor 0.3
l-Menthol 0.5
Purified water (enough to make the total amount 100 parts)

Example 11. Pack [Ingredients] Parts Dipropylene glycol 5.0
Polyoxyethylene (60) hydrogenated castor oil 5.0
Cetanol 3.0
Behenyl alcohol 3.0
Allantoin 0.1
Dipotassium glycyrrhizinate 0.1
Ammonium glycyrrhizinate 0.1
β-Glycyrrhetinic acid 0.1
Stearyl glycyrrhetinate 0.1
Salicylic acid 0.1
Tocopherol acetate 0.5
Tocopherol nicotinate 0.1
D-pantothenyl alcohol 0.3
Resorcinol 0.1
Sulfur 2.0
Estradiol 0.002
Hydrolyzate of Production Example 1 0.2
Xanthan gum 2.0
Polyglyceryl-6 myristate 1.0
Potassium cocoyl glutamate 1.0
Hydrogenated lecithin 3.0
Hydroxylated lecithin 3.0
Purified water (enough to make the total amount 100 parts)

Formulation Example 12. Hair Shampoo [Ingredients] Parts Sodium laureth sulfate 10.0
Glyceryl monostearate 1.0
Coconut oil fatty acid diethanolamide 2.0
Polyoxyethylene (40) hydrogenated castor oil 0.5
Benzalkonium chloride 1.0
Stearyl alcohol 2.0
Behenyl alcohol 2.0
Dimethicone 3.0
Hydrolyzate of Production Example 1 0.2
Allantoin 0.1
Dipotassium glycyrrhizinate 0.1
Salicylic acid 0.1
Sodium salicylate 0.1
Tocopherol acetate 0.1
Pyrithione zinc 0.3
Benzoic acid 0.2
Triclosan 0.2
Citric acid 0.1
Propylene glycol 2.0
Purified water (enough to make the total amount 100 parts)

Example 13. Hair conditioner [Ingredients] Parts Polyoxyethylene (10) hydrogenated castor oil 1.0
Distearyldimethylammonium chloride 1.5
Stearyltrimethylammonium chloride 2.0
Glyceryl 2-ethylhexanoate 1.0
Benzalkonium chloride 1.0
Cetanol 3.0
Stearyl alcohol 1.0
Hydrolyzate of Production Example 1 0.2
Salicornia herb extract 1.0
Black soybean hydrolysate 1.0
Allantoin 0.1
Isopropyl methylphenol 0.1
Dipotassium glycyrrhizinate 0.1
Salicylic acid 0.1
Sulfur 0.5
Alkylisoquinolinium bromide solution (75%) 0.06
Pyrithione zinc 0.3
Methylparaben 0.1
Triclosan 0.2
Resorcinol 0.1
Purified water (enough to make the total amount 100 parts)

Formulation Example 14. Cleansing cosmetic preparation [Ingredients] Parts Potassium cocoyl glycine 5.0
Glycerin 10.0
Glyceryl Caprylate 1.0
Sodium lauroyl aspartate 10.0
Hydrolyzate according to Production Example 1 0.2
Cetanol 3.0
Myristyl alcohol 3.0
Isopropyl methyl alcohol 0.1
Allantoin 0.1
Sulfur 0.5
Glycyrrhizic acid 0.1
Dipotassium glycyrrhizinate 0.1
Monoammonium glycyrrhizinate 0.1
β-Glycyrrhetinic acid 0.05
Stearyl glycyrrhetinate 0.1
Salicylic acid 0.2
Tocopherol acetate 0.2
Triclosan 0.1
Trichlorocarbanide 0.5
Trichlorohydroxydiphenyl ether 0.2
Concentrated benzalkonium chloride solution 50 0.2
Benzalkonium chloride 0.1
Purified water (enough to make the total amount 100 parts)

Formulation Example 15. Sheet Mask A sheet mask is obtained by impregnating a nonwoven fabric with the following ingredients.
[Ingredients] Parts Hydrolysate of Production Example 1 1.0
Glycerin 3.0
1,3-butylene glycol 2.0
L-Ascorbic acid 2-glucoside 2.0
Methylparaben 0.2
Citric acid 0.1
Sodium citrate 0.3
Xanthan gum 1.0
Water-soluble collagen 1.0
Sodium hyaluronate 1.0
Zostera extract 1.0
Potassium hydroxide (appropriate amount) Purified water (enough to make the total amount 100 parts)

Formulation Example 16. Serum [Ingredients] Parts Ethanol 2.0
Glycerin 5.0
1,3-butylene glycol 5.0
Methylparaben 0.1
Hyaluronic acid hydrolysate 1.0
Fermented lotus seeds 1.0
Lactic acid bacteria culture 1.0
Coconut oil fatty acid diethanolamide 4.0
Methylparaben 0.1
Citric acid 0.1
1,3-butylene glycol 2.0
Purified water (enough to make the total amount 100 parts)

Claims (3)

A protease activity enhancer containing, as an active ingredient, an enzymatic hydrolysate of polished rice or an extract of polished rice. The protease activity enhancer according to claim 1, wherein the polished rice is pigmented rice. The protease activity enhancer of claim 1 or claim 2, wherein the hydrolysate is a product treated with two or more proteolytic enzymes.