JP7246605B2 - Skin topical composition - Google Patents

Description

TECHNICAL FIELD The present invention relates to an external skin composition containing a plant-derived component as an active ingredient.

Conventionally, with aging, inactivation of cell proliferation and differentiation, decrease in hormone secretion, quantitative decrease in extracellular matrix components (collagen, laminin, etc.) that make up the skin, and muscle strength decrease due to decrease in muscle fibers, etc. It is known that skin aging phenomena (wrinkles, sagging, etc.) occur for the following reasons.

For the purpose of preventing and ameliorating the unhealthy cells and aging as described above, various active ingredients have been proposed, and cosmetics, foods and drinks, and pharmaceuticals containing these active ingredients have been put on the market. Examples include cell activating components such as α-hydroxycarboxylic acid, placental extract, and γ-amino-β-hydroxybutyric acid; extracellular matrix components such as collagen, elastin, hyaluronic acid or salts thereof; moisturizing agents such as urea; be done.

As described above, conventionally, cell activators and anti-aging agents have been proposed based on the mechanisms of aging and unhealthy cells. Problems exist in terms of effectiveness. Therefore, there is a demand for a novel active ingredient that overcomes these conventional problems.

As a result of intensive studies on the above problems of the prior art, the present inventors found that a plant containing cyclic adenosine monophosphate (cAMP) or its extract contains an enzyme (Adipose triglyceride lipase: ATGL) involved in the decomposition of fat cells. The present invention has been completed based on the new finding that by promoting or activating synthesis and promoting lipolysis, accumulation of fat and thereby improving skin sagging and wrinkles.

Conventionally, it has been known that cAMP or a plant extract containing cAMP has effects such as activating skin cells and promoting metabolism (Patent Document 1, Non-Patent Document 2). However, it is not known that high-concentration plant extracts containing cAMP promote the synthesis or activation of ATGL and promote lipolysis, thereby accumulating fat and thereby improving skin sagging and wrinkles. was not known.

JP-A-01-168605 Fragrance Journal extra edition No.6 (1986)

The present invention is a lipolysis promoter containing cyclic adenosine monophosphate (cAMP) as an active ingredient.
The present invention is a lipolysis promoter containing a plant containing cyclic adenosine monophosphate (cAMP) or an extract thereof as an active ingredient.
The present invention is an external skin composition for improving wrinkles and sagging, which contains cyclic adenosine monophosphate (cAMP), a plant containing the same, or an extract thereof as an active ingredient, and has lipolytic action.

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide an external skin composition for improving wrinkles and sagging, which is derived from a natural product, has excellent biosafety, and has a lipolytic action.

Preferred embodiments of the present invention are described in detail below.
In the present invention, cyclic adenosine monophosphate (cAMP) is a molecule synthesized from adenosine triphosphate (ATP) in which 3' and 5' of ribose and phosphate form a cyclic structure. may be a derivative thereof.

In the present invention, cyclic adenosine monophosphate (cAMP) or a derivative thereof may be a commercial product or an extract obtained from a natural product such as a plant by an extraction process. In addition, the plant extract may be subjected to a treatment such as separation, purification, concentration, etc. to increase the concentration of cyclic adenosine monophosphate (cAMP) or its derivative. Examples of plants containing cyclic adenosine monophosphate (cAMP) include jujube.

For the preparation of the extract, the plant to be extracted is washed with water in advance to remove foreign matter, if necessary, and then dried as it is or dried, and if necessary, finely chopped or pulverized, and then immersed or countercurrently applied. It can be carried out by contacting with an extraction solvent according to a conventional method such as an extraction method or a steam distillation method. Moreover, in the present invention, a supercritical extraction method may be employed.

Extraction solvents used for extract treatment include water; lower alcohols such as methanol, ethanol and propanol; polyhydric alcohols such as ethylene glycol, propylene glycol, 1,3-butylene glycol and glycerin; ethyl acetate and butyl acetate. , esters such as methyl propionate; ketones such as acetone and methyl ethyl ketone; ethers such as ethyl ether and isopropyl ether; Two or more kinds are mixed and used. In the present invention, it is more preferable to use a mixed solvent of two different polarities from the viewpoint of obtaining high concentrations of inulin and caffeic acid or derivatives thereof.

In preparation of the extract, there is no particular limitation on the pH of the extract, but it is generally preferred to be in the range of pH 3-9. The pH is adjusted by adding an alkalinity adjuster such as sodium hydroxide, sodium carbonate, or potassium hydroxide, or an acidity adjuster such as citric acid, hydrochloric acid, phosphoric acid, or sulfuric acid to the extraction solvent. be able to.

Extraction conditions such as extraction temperature and time vary depending on the type and pH of the solvent used, or the size of the plant material. In the case of the immersion method, the extraction temperature is in the range of 0 to 80°C. The extraction time is preferably in the range of 1 hour to 7 days in the case of cold extraction at 4°C, and in the range of 1 hour to 3 days in the case of medium temperature extraction around 40°C. In the case of high temperature extraction at 80° C., the time should be in the range of 1 hour to 24 hours. In the case of the immersion method, the bath ratio is generally 1 to 200 times, preferably 1 to 100 times the weight of the plant material.

The plant extract prepared as described above is further subjected to one or more of activated carbon treatment, ion exchange resin treatment, synthetic adsorbent, silica gel, and recrystallization treatment in combination to obtain cyclic adenosine monophosphate. It is preferable to prepare an extract containing a high concentration of Activated charcoal can be made from plant materials such as pine, bamboo, coconut shells, walnut shells, etc., as well as coal, petroleum, etc., and high-temperature carbonization using gases such as steam, carbon dioxide, and air for these raw materials. Activated carbon can be obtained by a physical method such as a chemical method such as a method using a chemical such as zinc chloride, and then heated to make it porous. Examples of ion exchange resins include weakly basic anion exchange resins, strongly acidic cation exchange resins, strongly basic ion exchange resins, and weakly acidic cation exchange resins. Examples of synthetic adsorbents include nonionic resins such as chelate resins, strongly acidic cation-adsorbing resins, styrene/divinylbenzene copolymers, and methacrylic acid ester polymers.

In the present invention, external skin compositions include cosmetics, quasi-drugs, and pharmaceuticals for external use. Red, white powder, facial cleanser, body shampoo, slimming agent, hair shampoo, soap, etc., also include hair growth agents, bath agents, etc., but are of course not limited to these.

In the present invention, when a plant extract containing cyclic adenosine monophosphate is used as an active ingredient, the concentration of cyclic adenosine monophosphate in the extract is 2.0 ppm or more, preferably 2.5 ppm or more.

In addition to the cyclic adenosine monophosphate according to the present invention or an extract containing the same, the external skin composition may contain components used in the external skin composition, such as oily components, surfactants (synthetic or natural products), Moisturizers, thickeners, antiseptic/bactericidal agents, powder components, ultraviolet absorbers, antioxidants, pigments, fragrances, and the like can be appropriately blended as necessary. In addition, as long as the effectiveness and characteristics of the extract, hydrolyzate, or fermented product according to the present invention are not impaired, other physiologically active ingredients may be used in combination.

Examples of oily components include olive oil, jojoba oil, castor oil, soybean oil, rice oil, rice germ oil, coconut oil, palm oil, cacao oil, meadowfoam oil, shea butter, tea tree oil, and avocado oil. , macadamia nut oil, bergamot oil, lavender oil, rose oil, bergamot oil, chamomile oil and other plant-derived squalane; animal-derived oils and fats such as mink oil and turtle oil; beeswax, carnauba wax, rice wax , Waxes such as lanolin; Hydrocarbons such as liquid paraffin, petrolatum, paraffin wax, and squalane; Fatty acids such as myristic acid, palmitic acid, stearic acid, oleic acid, isostearic acid, and cis-11-eicosenoic acid; Lauryl alcohol , cetanol, stearyl alcohol and other higher alcohols; isopropyl myristate, isopropyl palmitate, butyl oleate, 2-ethylhexylglyceride, synthetic esters and synthetic triglycerides such as higher fatty acid octyldodecyl (octyldodecyl stearate, etc.) mentioned.

Examples of surfactants include 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, poly 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 alkylphenyl ether sulfates, Anionic surfactants such as polyoxyethylene alkyl ether phosphates, α-sulfonated fatty acid alkyl ester salts, polyoxyethylene alkylphenyl ether phosphates; quaternary ammonium salts, primary to tertiary fatty amine salts, Cationic surfactants such as alkylbenzylammonium salts, alkylpyridinium salts, 2-alkyl-1-alkyl-1-hydroxyethylimidazolinium salts, N,N-dialkylmorphonium salts, polyethylenepolyamine fatty acid amide salts; N, N-dimethyl-N-alkyl-N-carboxymethylammoniobetaine, N,N,N-trialkyl-N-alkyleneammoniocarboxybetaine, N-acylamidopropyl-N',N'-dimethyl-N'- Amphoteric surfactants such as β-hydroxypropylammoniosulfobetaine and the like can be used.

Examples of emulsifiers and/or emulsifying aids include stevia derivatives such as enzyme-treated stevia, saponin or derivatives thereof, casein or salts thereof (such as sodium), complexes of sugar and protein, sucrose or its esters, lactose, soybean-derived Water-soluble polysaccharides, complexes of soybean-derived protein and polysaccharides, lanolin or its derivatives, cholesterol, stevia derivatives (stevia enzyme-treated products, etc.), silicates (aluminum, magnesium, etc.), carbonates (calcium, sodium, etc.), saponins and Derivatives thereof, lecithin and its derivatives (hydrogenated lecithin, etc.), lactic acid bacteria fermented rice, lactic acid bacteria fermented germinated rice, lactic acid bacteria fermented grains (barley, beans, cereals, etc.) and the like 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 sugars such as trehalose and raffinose. , mucopolysaccharides (e.g., hyaluronic acid and its derivatives, hyaluronic acid fermentation broth, 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 extracts, silane root (white and white) extracts, various amino acids and derivatives thereof.

Examples of thickening agents include alginic acid, agar, carrageenan, fucoidan, and other brown algae, green algae, or red algae-derived components; Birlan root (white) extract; pectin, polysaccharides such as aloe polysaccharide; tragacanth gum, locust bean gum, Gums such as xanthan gum and guar gum; Cellulose derivatives such as carboxymethyl cellulose, hydroxyethyl cellulose and hydroxypropyl cellulose; Polymers; hyaluronic acid and its derivatives; polyglutamic acid and its derivatives, and the like.

Antiseptic/bactericidal agents include, for example, urea; paraoxybenzoic acid esters such as methyl parahydroxybenzoate, ethyl parahydroxybenzoate, propyl parahydroxybenzoate, and butyl parahydroxybenzoate; phenoxyethanol, dichlorophen, hexachlorophene, chlorhexidine hydrochloride, benz chloride Ruconium, salicylic acid, ethanol, undecylenic acid, phenols, jamal (imidazodenyl urea), 1,2-pentanediol, propanediol, various essential oils, dry distillation of bark, fermented radish, sugar cane, plant-derived ingredients such as corn ethanol or 1,3-butylene glycol.

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. cereals (rice, wheat, corn, millet, etc.) powders, beans (soybeans, adzuki beans, etc.) powders, etc.

UV absorbers include, for example, ethyl para-aminobenzoate, ethylhexyl paradimethylaminobenzoate, amyl salicylate and derivatives thereof, 2-ethylhexyl para-methoxycinnamate, 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 butylhydroxyanisole, butylhydroxytoluene, propyl gallate, vitamin E and derivatives thereof (eg, vitamin E nicotinate, vitamin E linoleate, etc.).

Whitening agents include t-cycloamino acid derivatives, kojic acid and its derivatives, ascorbic acid and its derivatives, hydroquinone and its derivatives, ellagic acid and its derivatives, nicotinic acid and its derivatives, resorcinol derivatives, tranexamic acid and its derivatives, - methoxysalicylic acid potassium salt, magnolignan (5,5'-dipropyl-biphenyl-2,2'-diol), hydroxybenzoic acid and its derivatives, vitamin E and its derivatives, α-hydroxy acids, AMP (adenosine monophosphate) , adenosine monophosphate) and the like, and these may be blended singly or in combination.

Examples of the above kojic acid derivatives include kojic acid esters such as kojic acid monobutyrate, kojic acid monocaprate, kojic acid monopalmitate and kojic acid dibutyrate; kojic acid ethers; kojic acid sugar derivatives such as kojic acid glucoside; However, ascorbic acid derivatives include, 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, L-ascorbic acid-5-glucoside, ascorbyl tocopheryl maleate, ascorbyl tocopheryl potassium phosphate, myristyl 3-glyceryl ascorbate, caprylyl Ascorbic acid sugar derivatives such as 2-glyceryl ascorbic acid, 6-position acylated products of these ascorbic acid sugar derivatives (acyl group is hexanoyl group, octanoyl group, decanoyl group, etc.), L-ascorbic acid tetraisopalmitate, L- L-ascorbic acid tetrafatty acid esters such as ascorbic acid tetralaurate, 3-O-ethylascorbic acid, L-ascorbic acid-2-phosphate-6-O-palmitate sodium, glyceryl ascorbic acid or its acylation derivatives, ascorbic acid glycerol derivatives such as bisglyceryl ascorbic acid, aminopropyl L-ascorbate phosphate, hyaluronic acid derivatives of L-ascorbic acid, 3-OD lactose-L-ascorbic acid, isostearyl ascorbyl phosphate, etc. However, hydroquinone derivatives include arbutin (hydroquinone-β-D-glucopyranoside) and α-arbutin (hydroquinone-α-D-glucopyranoside), and tranexamic acid derivatives include tranexamic acid esters (eg, tranexamic acid lauryl ester, tranexamic acid hexadecyl ester, tranexamic acid cetyl ester or salts thereof), amides of tranexamic acid (e.g., tranexamic acid methylamide), etc., and resorcinol derivatives such as 4-n-butylresorcinol and 4-isoamylresorcinol. 2,5-dihydroxybenzoic acid derivatives such as 2,5-diacetoxybenzoic acid, 2-acetoxy-5-hydroxybenzoic acid, 2-hydroxy-5-propionyl xybenzoic acid and the like, nicotinic acid derivatives such as nicotinamide and benzyl nicotinate, and α-hydroxy acids such as lactic acid, malic acid, succinic acid, citric acid and α-hydroxyoctanoic acid. .

Examples of physiologically active ingredients include placenta extract, saxifrage extract, saxifrage extract, perilla extract, rice bran extract or its hydrolyzate, white mustard extract or its hydrolyzate, fermented white mustard, peony extract or hydrolyzate thereof, lactic acid bacteria fermented rice, purple extract, lotus seed extract or hydrolyzate thereof, fermented lotus seed, extract or hydrolyzate thereof, hydrolyzate of adlay, fermented adlay seed, Royal jelly fermented product, sake lees extract or ceramide contained therein, sake lees fermented product, Pandanus amaryllifolius extract, Arcangelicia flava extract, chamomile extract and the like. In addition, coral grass extract, rice leaf extract or its hydrolyzate, eggplant (lotus, long eggplant, Kamo eggplant, rice eggplant, etc.) extract or its hydrolyzate, apricot fruit extract, eucheuma cornflower, etc. seaweed extract, marine flowering plant extract such as eelgrass, fermented soymilk, jellyfish water, rice extract or its hydrolyzate, fermented rice extract, germinated rice extract or its hydrolyzate, fermented germinated rice , black soybean extract or its hydrolyzate, damask rose flower extract, bamboo shoot skin extract, linoleic acid and its derivatives or processed products (e.g., liposomal linoleic acid, etc.), animal or fish-derived collagen and Its derivatives, elastin and its derivatives, glycyrrhizic acid and its derivatives (dipotassium salt, etc.), t-cycloamino acid derivatives, vitamin A and its derivatives, allantoin, diisopropylamine dichloroacetate, γ-amino-β-hydroxybutyric acid, gentian extract , licorice extract, carrot extract, panax ginseng extract or its fermented product, red ginseng extract, honey kelp extract, loofah extract, sea lettuce extract, peach extract, peach kernel extract, kiwi extract, juazeiro extract, Pau d'Arco bark extract, day lily extract or fermented product, red wormwood extract, cherimoya extract, mango extract, mangosteen extract, funori extract, oolong tea extract, red fuki extract, cilantro extract, sansho pericarp Or seed coat extract or hydrolyzate, safflower flower extract, Casablanca extract, sweet potato extract or its fermented product, guava leaf extract, Houttuynia cordata extract, Banpeiyu extract, aloe extract, fig flower extract , apple extract, white asparagus extract, etc.

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

Production example 1. Preparation of cAMP-containing plant extract (1)
Jujube fruits belonging to the genus Jujube of the family Buckthornaceae were dried, and 2 kg of the dried product was added with 20 kg of purified water and immersed at 40°C. This was filtered to obtain 18 kg of a transparent brown jujube fruit water extract (solid concentration: 5.0%). Cyclic adenosine monophosphate was concentrated in the obtained water extract of jujube fruit using a styrene-divinylbenzene-based synthetic adsorbent. Purified water and 1,3-butylene glycol were added to 150 g of the obtained concentrate and mixed, and then insoluble matter was filtered out to obtain 8.9 kg of pale brown and transparent jujube fruit extract (solid concentration: 1.1%).

Production example 2. Preparation of cAMP-containing plant extract (2)
Jujube fruits belonging to the genus Jujube of the family Buckthornaceae were dried, and 2 kg of the dried product was added with 20 kg of purified water and immersed at 40°C. This was filtered to obtain 18 kg of a transparent brown jujube fruit water extract (solid concentration: 5.0%). A styrene-divinylbenzene-based synthetic adsorbent was added to the obtained water extract of jujube fruit, and the mixture was gently stirred at normal temperature, and the adsorbent was recovered after treatment. A 70% ethanol solution was added to this adsorbent, and after gentle stirring, the adsorbent was removed to obtain a brown transparent eluate. Insoluble matter was filtered from the obtained eluate to obtain 8.9 kg of pale brown and transparent jujube fruit extract (solid concentration: 1.1%).

Production example 3. Preparation of cAMP-containing plant extract (3)
Jujube fruits belonging to the genus Jujube of the family Buckthornaceae were dried, and 2 kg of the dried product was added with 20 kg of purified water and immersed at 40°C. This was filtered to obtain 18 kg of a transparent brown jujube fruit water extract (solid concentration: 5.0%). A styrene-divinylbenzene-based synthetic adsorbent was added to the obtained water extract of jujube fruit, and the mixture was gently stirred at normal temperature, and the adsorbent was recovered after treatment. A 70% ethanol solution was added to this adsorbent, and after gentle stirring, the adsorbent was removed to obtain a brown transparent eluate. The resulting eluate was concentrated by distillation to obtain a concentrate. Insoluble matter was removed from the concentrate by filtration to obtain 8.9 kg of pale brown and transparent jujube fruit extract (solid concentration: 1.1%).

Comparative production example 1. Preparation of plant extract Fruits of Jujube of the genus Jujube of the family Buckthornaceae were dried, and 2 kg of the dried product was added with 20 kg of a 90% ethanol solution and immersed at 40°C. This was filtered to obtain 18 kg of a transparent brown jujube fruit water extract (solid concentration: 5.3%). This was diluted with purified water to a solid content concentration of 1.1% and used as a comparative sample in the following tests.

Comparative production example 2. Preparation of Plant Extract Fruits of jujube of the genus Jujube of the family Buckthornaceae were dried, and 2 kg of the dry matter was added with 20 kg of a 70% 1,3-butylene glycol solution and immersed at 40°C. This was filtered to obtain 18 kg of a transparent brown jujube fruit aqueous extract (solid concentration: 5.1%). This was diluted with purified water so that the solid content concentration was 1.1%, and subjected to the following tests.

Formulation example 1. Lotion [Ingredients] Part Olive oil 1.0
Polyoxyethylene (5.5) cetyl alcohol 5.0
Butylparaben 0.1
Extract of Production Example 1 5.0
Ethanol 5.0
Glycerin 5.0
1,3-butylene glycol 5.0
Potassium hydroxide Appropriate amount Purified water Amount to make the total amount 100 parts Perfume Appropriate amount

Formulation example 2. Lotion A lotion was obtained in the same manner as in Formulation Example 1, except that 5.0 parts of the extract of Production Example 2 was used in place of the extract of Production Example 1 contained in Formulation Example 1.

Formulation example 3. Lotion A lotion was obtained in the same manner as in Formulation Example 1, except that 5.0 parts of the extract of Production Example 3 was used in place of the extract of Production Example 1 contained in Formulation Example 1.

Formulation example 4. Emulsion [Ingredients] Part Liquid paraffin 6.0
Hexalane 4.0
Jojoba oil 1.0
Polyoxyethylene (20) sorbitan monostearate 1.0
Lipophilic glyceryl stearate 1.0
Soy lecithin 1.5
Extract 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
Carboxymethyl cellulose 0.3
Sodium hyaluronate 0.01
Purified water Amount that makes the total amount 100 parts Fragrance Appropriate amount

Formulation example 5. Milky Lotion A milky lotion was obtained in the same manner as in Formulation Example 4, except that 3.0 parts of the extract of Production Example 2 was used in place of the 3.0 parts of the extract of Production Example 1.

Formulation example 6. Milky Lotion A milky lotion was obtained in the same manner as in Formulation Example 4, except that 3.0 parts of the extract of Production Example 3 was used in place of the 3.0 part of the extract of Production Example 1.

Formulation example 7. Milky lotion A milky lotion is obtained in the same manner as in Formulation Example 4, except that 2.0 parts of L-ascorbic acid-2-glucoside and 3.0 parts of arbutin are used in place of 0.5 parts of potassium hydroxide. rice field.

Formulation example 8. Milky lotion A milky lotion is prepared in the same manner as in Formulation Example 4, except that 2.0 parts of L-ascorbic acid-2-glucoside and 2.0 parts of tranexamic acid are used instead of 0.5 parts of potassium hydroxide. Obtained.

Formulation example 9. Milky lotion Milky lotion in the same manner as in Formulation Example 4, except that 2.0 parts of L-ascorbic acid-2-glucoside and 3.0 parts of nicotinamide are used instead of 0.5 parts of potassium hydroxide. got

Formulation example 10. Lotion [Ingredients] Part Extract of Production Example 1 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
Xanthan gum 0.1
Fragrance Appropriate amount Potassium hydroxide Appropriate amount Purified water Amount to make the total amount 100 parts

Formulation example 11. Essence [ingredient] part ethanol 2.0
Glycerin 5.0
1,3-butylene glycol 5.0
Methylparaben 0.1
Hyaluronic acid 0.1
Extract of Production Example 1 5.0
Citric acid 0.3
Sodium citrate 0.6
Purified water Amount that makes the total amount 100 parts

Formulation example 12. Essence An essence was obtained in the same manner as in Formulation Example 11, except that 5.0 parts of the extract of Production Example 1 was replaced with 5.0 parts of the extract of Production Example 1.

Example 13. Liquid foundation [ingredients] 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
Extract of Production Example 1 5.0
Carboxymethylcellulose sodium 0.2
Bentonite 0.5
Propylene glycol 4.0
Triethanolamine 1.1
Methylparaben 0.1
Purified water Amount to make the total amount 100 parts Titanium oxide 8.0
Talc 4.0
Appropriate amount of coloring pigment

Formulation example 14. Body Shampoo [Ingredients] Part N-lauroylmethylalanine sodium 25.0
Coconut fatty acid potassium liquid (40%) 26.0
Coconut fatty acid diethanolamide 3.0
Methylparaben 0.1
Extract of Production Example 3 5.0
1,3-butylene glycol 2.0
Purified water Amount that makes the total amount 100 parts

Formulation example 15. Hair restorer [Ingredients] Part dipotassium glycyrrhizinate 0.1
Mononitroguaiacol sodium 0.02
Pyridoxine hydrochloride 0.03
l-menthol 0.8
Tamasakitsutsurafuji root extract 0.3
Brown algae extract 0.3
Panax ginseng extract 0.3
Gentian extract 2.0
Extract of Production Example 2 3.5
Trimethylglycine 0.5
Lactic acid 0.2
1,3-butylene glycol 10.0
Phenoxyethanol 0.2
Polyoxyethylene hydrogenated castor oil 0.4
L-Arginine Appropriate amount Ethanol 20.0
Purified water Amount that makes the total amount 100 parts

Formulation example 16. Hair Shampoo [Ingredient] Part Sodium N-coconut oil fatty acid methyl taurate 10.0
Sodium polyoxyethylene (3) alkyl ether sulfate 20.0
Lauryldimethylaminoacetate betaine 10.0
Coconut fatty acid diethanolamide 4.0
Methylparaben 0.1
Citric acid 0.1
Extract of Production Example 3 2.0
1,3-butylene glycol 2.0
Purified water Amount that makes the total amount 100 parts

Example 17. Hair conditioner [Ingredients] Part polyoxyethylene (10) hydrogenated castor oil 1.0
Distearyldimethylammonium chloride 1.5
Stearyltrimethylammonium chloride 2.0
Glyceryl 2-ethylhexanoate 1.0
Cetanol 3.2
Stearyl alcohol 1.0
Methylparaben 0.1
Extract of Production Example 2 2.0
1,3-butylene glycol 5.0
Purified water Amount that makes the total amount 100 parts

Test example 1. Quantitative analysis _of cyclic adenosine monophosphate (1) Preparation of cyclic adenosine monophosphate standard undiluted solution About 10 mg of cyclic adenosine monophosphate [ _C10H12N5O6P : 329.21 (97% or higher purity)] was precisely _weighed and _purified . Add water to make exactly 100 mL, and use this as the cyclic adenosine monophosphate standard stock solution.
(2) Preparation of calibration curve Accurately take 1 mL, 5 mL, and 10 mL of the cyclic adenosine monophosphate standard undiluted solution, add purified water to make exactly 100 mL, and use the standard solution exclusively for calibration. Take 10 μL of the standard solution dedicated for calibration, and perform the test by liquid chromatography under the above test conditions. A calibration curve is created by plotting the peak area of the resulting chromatogram on the vertical axis and the concentration of cyclic adenosine monophosphate (mg/mL) on the horizontal axis.
(3) Preparation of sample solution Accurately weigh about 1 g of the extracts of Production Examples 1 to 3, add water to make exactly 10 mL, and use this as the sample solution.
(4) Quantitative analysis Accurately take 10 µL of the sample solution and perform the test by liquid chromatography under the following test conditions. The peak area of the resulting chromatogram is determined, and the amount of cyclic adenosine monophosphate is calculated from a calibration curve prepared in advance.
Amount of cyclic adenosine monophosphate in this product: S (ppm) = Amount of cyclic adenosine monophosphate obtained from the calibration curve (mg/mL) x (10/amount of product collected (g)) x 1000/0.97
<Detection conditions>
Detector: Ultraviolet absorption photometer (measurement wavelength: 254 nm)
Column: A stainless steel tube with an inner diameter of 4.6 mm and a length of 150 mm is packed with octadecylsilanized silica gel for liquid chromatography.
Column temperature: Constant temperature around 30°C Mobile phase: Add 3.12 g of sodium dihydrogen phosphate dihydrate to 980 mL of purified water, adjust the pH to 2.5 with phosphoric acid, and make up to 1000 mL with purified water.
Flow rate: 1.0 mL/min (retention time of cyclic adenosine monophosphate approx. 11 min)

Table 1 shows the measurement results of Test Example 1.
[Table 1]

Test example 2. Evaluation test of ATGL synthesis promotion effect
Mouse fibroblasts (3T3-L1) were seeded at 1.5×10 ⁴ cells/well in a 96-well microplate containing 10% FBS-containing Dulbecco's Modified Eagle's Minimal Essential Medium (DMEM: Nissui Pharmaceutical Co., Ltd.) and incubated at 37°C. , After pre-culture for 3 days under the condition of 5.0% CO ₂ , differentiation induction medium [10% FBS, 0.5 mM 3-isobutyl-1-methylxanthine (IBMX), 0.25 μM dexamethasone (DEX) and 1.1 μg/mL After adding DMEM mixed with insulin], the cells were further cultured for 2 days. After replacing the culture medium with DMEM mixed with 1.1 μg/mL insulin and culturing for another 2 days, the extracts of Production Examples 1 to 3 and Comparative Production Examples 1 and 2 were used as sample solutions and adjusted to a specified concentration. 10% FBS-containing DMEM was added, and the cells were further cultured under the same conditions for 3 days. Here, the sample solution was adjusted so that the final concentration of each extract as a solution was 0.5% and 1.0%. Immunodetection with ATGL antibody was then performed. That is, after washing with PBS(-), the cells were treated with 15% neutral buffered formalin for 30 minutes, fixed, permeabilized with 0.5% Triton X-100 solution for 1 hour, and 5-fold diluted Blocking One P (Nacalai Tesque). ) solution for 2 hours for blocking, then ATGL antibody was added and allowed to stand at room temperature for 1 hour. After that, the plate was washed with PBS(-), a fluorescently labeled secondary antibody was added, and the plate was allowed to stand in the dark for a certain period of time. After washing with PBS(-), fluorescence intensity was measured. First, the fluorescent label of the secondary antibody (Alexa Fluor544) was measured at Ex = 544 nm and Em = 590 nm (fluorescent microplate reader [Fluoroscan Ascent, Thermo Fisher Scientific)], then DNA was stained with Hoechst33342, Ex = 355 nm and Em = 460 nm. The degree of ATGL formation was obtained by dividing the fluorescence intensity of Alexa Fluor544 in each test plot by the fluorescence intensity of Hoechst33342. The same operation as above was performed for the case of no sample addition (control) in which 30BG was added instead of the sample solution, and the relative value of the degree of ATGL formation when each sample was added to the degree of ATGL formation obtained here was obtained, The amount of ATGL produced (%).

Table 2 shows the results of Test Example 2.
[Table 2]

As shown in Table 2, it was confirmed that the extract containing cyclic adenosine monophosphate according to the present invention has a remarkably excellent effect of promoting APGL synthesis.

Test example 3. Evaluation test of ATGL activation effect
Mouse fibroblasts (3T3-L1) were seeded at 4×10 ⁶ cells/flask in 80 cm ² flask containing 10% FBS-containing Dulbecco's Modified Eagle's Minimum Essential Medium (DMEM: Nissui Pharmaceutical Co., Ltd.). After pre-culturing for 3 days under 5.0% CO ₂ conditions, the extracts of Production Examples 1 to 3 and Comparative Production Examples 1 to 2 were used as sample solutions and adjusted to a specified concentration in a differentiation induction medium [10% FBS , 0.5 mM 3-isobutyl-1-methylxanthine (IBMX), 0.25 μM dexamethasone (DEX), and DMEM mixed with 1.1 μg/mL insulin], and cultured for an additional 2 days. Here, the sample solution was adjusted so that the final concentration of each extract as a solution was 2.0%. After culturing for 2 days, the culture medium was replaced with DMEM containing each sample solution at the above concentrations and mixed with 1.1 μg/mL insulin, and cultured for another 5 days. After washing the cells with PBS(-), 0.5g/L Trypsin/0.53mmol/L EDTA solution (Nacalai Tesque, Inc.) was added and allowed to stand at 37°C for 5 minutes to detach the cells from the flask. A suspension was obtained. The cell suspension was collected and disrupted according to the Lipoprotein Lipase (LPL) Activity Assay Kit (CELL BIOLABS), and ATGL activity was measured. Considering the effect of the sample solution on stimulating the cells and inhibiting proliferation, the amount of protein in the cell lysate was measured by the Bradford method, and the ATGL activity was measured after preparing the proteins. 30% 1,3-butylene glycol solution was added instead of the sample solution, and no sample was added (control). The relative value of was determined and used as the ATGL activation rate (%).

Table 3 shows the results of Test Example 3.
[Table 3]

As shown in Table 3, it was revealed that the extract containing cyclic adenosine monophosphate according to the present invention has a remarkably superior ATGL activating effect.

Test example 4. Evaluation test of lipid droplet decomposition promotion effect
Mouse fibroblasts (3T3-L1) were seeded at 1.5×10 ⁴ cells/well in a 96-well microplate containing 10% FBS-containing Dulbecco's Modified Eagle's Minimal Essential Medium (DMEM: Nissui Pharmaceutical Co., Ltd.) and incubated at 37°C. , after pre-culture for 3 days under 5.0% CO ₂ conditions, differentiation induction medium [10% FBS, 0.5 mM 3-isobutyl-1-methylxanthine (IBMX), 0.25 μM dexamethasone (DEX) and 1.1 μg/mL insulin DMEM mixed with] was added, and cultured for an additional 2 days. After replacing with DMEM mixed with 1.1 μg/mL insulin and culturing for another 5 days, the extracts of Production Examples 1 to 3 and Comparative Production Examples 1 and 2 were used as sample solutions and adjusted to a specified concentration of 10%. FBS-containing DMEM was added, and the cells were further cultured under the same conditions for 2 days. Here, the sample solution was adjusted so that the final concentration of each extract as a solution was 0.5% and 1.0%. Then, intracellular lipid droplets were stained using Nile Red (Wako Pure Chemical Industries, Ltd.). That is, after washing with PBS(-), the cells were fixed by treating with a 10% formalin aqueous solution for 10 minutes. After that, it was washed with PBS(-), added with Nile Red acetone solution, and allowed to stand at room temperature in a dark place for 5 minutes. After washing with PBS (-), Ex = 544 nm, Em = 590 nm were measured using a fluorescence microplate reader (fluoroscan Ascent, manufactured by Thermo Fisher Scientific), and this was used as the residual amount of lipid droplets. After fluorescence measurement, DNA staining with Hoechst33342 was performed, and Ex=355 nm and Em=460 nm were measured. The accumulated amount of lipid droplets was determined by dividing the absorbance at ABS540 nm of each test group by the fluorescence intensity of Hoechst33342. In place of the sample solution 30% 1,3-butylene glycol solution was added in the case of no sample addition (control), the same operation as above was performed, and the amount of lipid droplets remaining obtained here at the time of each sample addition The relative value of the residual lipid droplets was determined and used as the lipid droplet decomposition amount (%).

Table 4 shows the results of Test Example 4.
[Table 4]

As shown in Table 4, it was revealed that the extract containing cyclic adenosine monophosphate according to the present invention has a remarkably excellent lipid droplet decomposition promoting effect.

Test example 5. Monitor test (improvement of sagging cheeks)
Three healthy subjects washed their faces with a commercially available face wash, and after resting for 15 minutes in an examination room with a room temperature of 24°C and a humidity of 50-60%, points were set on the subjects' faces. The points are set as follows. A point P1 was set 1 cm from the corner of the eye, and a point P2 was set 2 cm from the corner of the mouth. Three points were set at equal intervals between P1 and P2, and P3, P4, and P5 were set from the top. P6 is the perpendicular intersection of lines drawn from P4 toward the center of the face and downward from the center of the eye. Of these, P6 was sag A and P5 was sag B. At points A and B, we took pictures of the subjects lying down and sitting down with a digital camera. After the initial exposure, the subjects were half-faced with a milky lotion containing the placebo and the extract of Preparation Example 3 twice a day, morning and evening. One month after the start of the test, images were taken under the same conditions. Before and after the test, the moving distance from the reference point was calculated by image processing, and changes in the moving distance before and after the test were confirmed.

Table 5 shows the results of Test Example 5.
[Table 5]

As shown in Table 5, it was confirmed that the extract according to the present invention improved cheek sagging. Thus, the present invention can provide an external composition for skin that has an effect of improving slackness, wrinkles, skin tightening, and nasolabial folds.

Claims (1)

An external skin composition for improving sagging and wrinkles on cheeks, comprising as an active ingredient a jujube fruit extract containing 2 ppm or more of cyclic adenosine monophosphate .