JP2011016726A - Skin care composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a skin care composition that suppresses skin wrinkle formation by taking care of the skin, maintains skin firmness and elasticity and keeps a young skin state.SOLUTION: The skin care composition comprises (1) an extract from Phaseolus atropurpureus, (2) an extract from royal jelly and/or an extract from ginseng and (3) at least one kind of a lipid-soluble vitamin selected from vitamin A or its fatty acid ester, vitamin D, vitamin E, vitamin B2 fatty acid ester, vitamin B6 fatty acid ester, vitamin C fatty acid ester, pantothenol and niacin as active ingredients. The components (1) and (2) are present in a hydrophilic phase (W phase) and the component (3) is present in a lipophilic phase (O phase).

Description

  The present invention relates to an external composition for skin having a reinforcing and protective effect on the basement membrane involved in the homeostatic function of the epidermis.

  Skin structure is roughly divided into the stratum corneum, epidermis, basement membrane, dermis, and subcutaneous tissue. The basement membrane exists at the boundary between the epidermis and the dermis, and its functions are diverse, factors such as adhesion of the epidermis to the dermis, determination of the polarity of the epidermis, control of epidermal differentiation and proliferation, and factors produced by dermal cells It is involved in the control of nutrient supply derived from blood and blood components. Therefore, the epidermal basement membrane plays an extremely important role in maintaining the structure and homeostatic function of the skin.

  The main skeleton of the basement membrane has a network structure composed of type IV collagen, and further has laminin 5 and type VII collagen. Various glycoproteins mainly composed of laminin 5 exist at the boundary between the basement membrane and the epidermis and connect the basement membrane and the epidermis, and type VII collagen is responsible for the connection between the basement membrane and the dermis. In young skin, the basement membrane works to maintain the constant interaction between the epidermis and dermis, ensuring moisture retention, flexibility, elasticity, etc., and the skin looks fresh and supple. Maintained. However, when there is an influence of certain external factors such as ultraviolet irradiation, drastic air drying, excessive skin washing, etc., or when aging progresses, type IV collagen, which is the main component of the basement membrane, and laminin 5 undergoes degradation and alteration, and the basement membrane structure is destroyed. As a result, the skin's moisturizing function and elasticity are lowered, and the keratin begins to exfoliate abnormally, so the skin loses its tension and gloss, and exhibits aging symptoms such as roughness and wrinkles. In this way, changes accompanying skin aging, that is, wrinkles, dullness, disappearance of texture, decreased elasticity, etc. involve structural changes such as a decrease in basement membrane components, basement membrane multiplexing, and tearing. Yes.

  Therefore, maintaining the structure of the epidermal basement membrane in a good state is also important from the viewpoint of preventing skin health and future aging, so the importance of skin basement membrane care as a new skin care is increasing. . For this reason, by promoting the production of type IV collagen and laminin 5, the remodeling of the basement membrane structure is induced, and the activity of MMPs known as enzymes that degrade type IV collagen and laminin 5 is inhibited. Development of a substance that suppresses the decrease in the basement membrane component and the structural change of the basement membrane and improves the skin function is desired.

  Examples of the type IV collagen production-promoting action include hydrolyzed casein, pre-extract, beech bud extract, erythrina extract, soluble eggshell membrane, cuckoo extract and western kizuta extract (Patent Document 1), saponin or sapogenol (Patent Document) 2) etc. are known. Examples of those having a laminin 5 production promoting action include, for example, a cuckoo extract, a licorice extract or a flavonoid fraction thereof, a cypress extract, a dicytic extract, a lobster extract, a koroha extract, and a whey extract ( Patent Document 3) Onionis extract, Merirot extract, bean extract, azuki bean extract (Patent Document 4), soybean extract (Patent Document 5) and the like are known. Furthermore, what has a matrix metalloprotease inhibitory action is known, for example, the black reishi extract (patent document 6), the extract from chestnut astringent skin fermented product (refer patent document 7), etc. Furthermore, a type IV collagen production promoter, laminin 5 production promoter, and a matrix metalloprotease inhibitor that contains an extract from Azuki beetle as an active ingredient have been proposed (Patent Document 8).

JP 2004-18471 A JP 2002-516837 A JP 2003-137767 A JP 2003-137768 A JP 2004-217618 A JP 2005-298391 A JP 2004-352634 A JP 2007-217352 A

  As a result of repeating various experiments on the above-described type IV collagen production promoter, laminin 5 production promoter, etc., the present inventors, as the type IV collagen production promoter, are excellent extracts from black azuki bean, In addition to the ability to promote type IV collagen production and laminin 5 production, in addition to the ability to proliferate fibroblasts that maintain the elasticity of the skin, a combination of several specific active ingredients is used in addition to black azuki bean. I found that it was necessary. The fibroblast proliferating ability is the ability to proliferate fibroblasts that produce dermis components present in the dermis reticulate layer, and the elasticity and youthfulness of the skin can be obtained by maintaining and improving the fibroblasts.

  For this reason, with the above materials, it is difficult to maintain the youthful skin condition by suppressing skin wrinkle formation by caring for the skin (particularly the basement membrane), maintaining the firmness and elasticity of the skin. there were.

  Accordingly, an object of the present invention is to provide an external composition for skin that can suppress the formation of wrinkles in the skin by caring for the skin, maintain the firmness and elasticity of the skin, and maintain a youthful skin condition. To do.

The above purpose is
As an active ingredient
(1) an extract from black-spotted azuki bean,
(2) Extract from royal jelly and / or carrot extract, and (3) Vitamin A or its fatty acid ester, Vitamin D, Vitamin E, Vitamin B2 fatty acid ester, Vitamin B6 fatty acid ester, Vitamin C fatty acid At least one fat-soluble vitamin selected from esters, pantothenol and niacin;
A composition for external application to the skin, comprising:
Can be achieved.

Preferred embodiments of the external composition for skin of the present invention are listed below.
(I) (1) An extract from black-spotted azuki bean has type IV collagen production ability and laminin 5 production ability. This has a reinforcing and protective effect on the basement membrane involved in the homeostatic function of the epidermis.
(Ii) (2) The extract from royal jelly and / or carrot extract has fibroblast proliferation ability. In addition, the extract from royal jelly has the ability to produce type IV collagen and the carrot extract has the ability to produce laminin 5.
(Iii) The above components (1) and (2) are present in the hydrophilic phase (W phase), and the component (3) is present in the lipophilic phase (O phase). Thereby, stability of an emulsion and the permeability to skin improve.
(Iv) The average particle diameter of the lipophilic phase (O phase) containing the component (3) is 100 nm or less. This further improves the stability of the emulsion and the permeability to the skin.
(V) The hydrophilic phase (W phase) contains glycerin. Thereby, stability of an emulsion and the permeability to skin improve.
(Vi) Does not contain parabens, synthetic dyes and synthetic fragrances. Thereby, skin irritation is reduced.
(Vii) It is in the form of an emulsion, lotion, cream or gel (gelled emulsion).

  In the composition for external use of the skin of the present invention, (1) an extract from black azuki bean, (2) an extract from royal jelly and / or an extract from carrot, and (3) a specific fat-soluble vitamin are used. The By applying such a composition containing the components (1) to (3) to the skin, it suppresses the formation of wrinkles on the skin, maintains the firmness and elasticity of the skin, and maintains the youthful skin condition. Can be maintained. Particularly, (1) to (3) emulsified compositions in which the components (1) and (2) are present in the hydrophilic phase (W phase) and the component (3) is present in the lipophilic phase (O phase). By applying to the skin, the above effects can be further improved. This is because the ability to produce type IV collagen and the ability to produce laminin 5 are greatly improved and the ability to proliferate fibroblasts that give elasticity to the skin can be improved by simultaneously using the above three or four components. Or it is thought that the fall of these abilities can be suppressed. Therefore, the composition for external use of the present invention is capable of suppressing the formation of wrinkles on the skin by caring for the skin, maintaining the firmness and elasticity of the skin, and maintaining the youthful skin condition. .

  The skin external composition of the present invention will be described in detail below.

  In the composition for external use of the skin of the present invention, as an active ingredient thereof, (1) an extract from black-spotted azuki bean, (2) an extract from royal jelly and / or an extract from carrot, and (3) specific fat solubility Vitamins are used.

  The black-tailed azuki bean (Phaseolus atropurpureus) used in the present invention is a perennial vine belonging to the genus of the leguminous bean family that is native to North America and mainly distributed in Kyushu, Okinawa, etc. in Japan. Can be obtained. As the extraction raw material, for example, parts such as a trunk, an above-ground part, a root part, a flower part, and a whole plant can be used, but a trunk part is preferable. Kurobanatsuruzuki is called Siratro in English.

  In the present invention, the extract (1) from black azuki bean is an extract obtained by using black azuki bean as an extraction raw material, a diluted or concentrated solution of this extract, and a dry obtained by drying the extract. Or any of these crudely purified products or purified products.

  The black azuki bean extract is considered to have type IV collagen producing ability (promoting action), laminin 5 producing ability, and further matrix metalloprotease inhibitory action, and is obtained by an extraction method generally used for plant extraction.

  For example, after drying the black azuki bean, it is pulverized as it is or using a crusher, and subjected to extraction with an extraction solvent, whereby an extract having type IV collagen production ability and laminin 5 production ability can be obtained. Drying may be performed in the sun or using a commonly used dryer. Moreover, after performing pretreatment, such as degreasing, with a nonpolar solvent such as hexane, it may be used as an extraction raw material. By performing pretreatment such as degreasing, extraction with a polar solvent can be performed efficiently.

  Examples of the extraction solvent include polar solvents such as water and hydrophilic organic solvents. These can be used alone or in combination of two or more. The extraction is preferably performed at room temperature or a temperature not higher than the boiling point of the solvent.

  As water that can be used as the extraction solvent, pure water, tap water, well water, mineral water, spring water, fresh water, and the like, as well as those treated in various ways can be used. Examples of such treatment include purification, heating, sterilization, filtration, ion exchange, osmotic pressure adjustment, and buffering.

  Examples of hydrophilic organic solvents that can be used as the extraction solvent include lower aliphatic alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; 3 to 5 lower carbon atoms such as acetone and methyl ethyl ketone. Aliphatic ketones: C2-C5 polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, glycerin and the like can be mentioned. Methanol, ethanol, isopropyl alcohol, methyl ethyl ketone, and 1,3-butylene glycol are preferable, and ethanol and 1,3-butylene glycol are particularly preferable.

  When using the liquid mixture of 2 or more types of polar solvents as an extraction solvent, the mixing ratio can be adjusted suitably. For example, when using a liquid mixture of water and a lower aliphatic alcohol, it is preferable to mix 1 to 90 parts by weight of a lower aliphatic alcohol with respect to 10 parts by weight of water. When using a mixed solution, it is preferable to mix 1 to 40 parts by mass of a lower aliphatic ketone with 10 parts by mass of water, and when using a mixed solution of water and a polyhydric alcohol, water 10 It is preferable to mix 10 to 90 parts by mass of polyhydric alcohol with respect to parts by mass.

  The extraction treatment is not particularly limited as long as the soluble component contained in the extraction raw material can be eluted in the extraction solvent, and can be performed according to a conventional method. For example, the extraction raw material is immersed in 5 to 15 times the extraction solvent (mass ratio) of the extraction raw material, the soluble components are eluted at room temperature or under reflux, and then filtered to remove the extraction residue. A liquid can be obtained. The obtained extract is diluted, concentrated, dried, purified, etc. according to a conventional method to obtain a diluted or concentrated solution of this extract, a dried product of the extract, or a crudely purified product or purified product thereof. May be applied.

  You may perform pressure control for raising extraction efficiency.

  Purification can be performed by, for example, activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, or the like. The obtained extract can be used as it is as an active ingredient having type IV collagen production ability and laminin 5 production ability, but a concentrated solution or a dried product is easier to use.

  Since the black azuki bean extract has a unique smell and taste, it can be purified for the purpose of decolorization, deodorization, etc. within a range that does not reduce its physiological activity. Since it is not used in a large amount when blended into a material, etc., it may be used unpurified.

  The blending amount of the black azuki bean extract with respect to the external composition for skin of the present invention is 0.001 to 1% by mass (nonvolatile content 0.00001 to 0.01% by mass), particularly 0.01 to 0.5% by mass (nonvolatile content). 0.0001 to 0.005 mass%) is preferable. The index component of the black-throated azuki bean extract is kaempferol glycoside and flavonoid.

  Since the extract of Azuki beetle obtained as described above has the ability to produce type IV collagen and laminin 5, it has a reinforcing and protective effect on the basement membrane involved in the homeostatic function of the epidermis. In addition, it is considered to have fibroblast proliferation ability. Therefore, it is used as an active ingredient of the external composition for skin of the present invention.

  The active ingredient of the composition for external use of the present invention includes (2) an extract from royal jelly and / or an extract from carrot, and (3) a specific fat-soluble vitamin, in addition to the black azuki bean extract. The

  Royal Jelly (2) is a secretion from the pharyngeal gland of a young worker bee in the European honey bee family (Apis melifera L.), which is fed to the queen bee larva and adult queen bee. is there. As the royal jelly extract, an ethanol extract of royal jelly is generally used.

  You may perform temperature control and pressure control for raising extraction efficiency.

  The blending amount of the extract from royal jelly with respect to the external composition for skin of the present invention is 0.001 to 1% by mass (non-volatile content 0.000005 to 0.005% by mass), particularly 0.01 to 0.1% by mass (non-volatile content). 0.00005 to 0.0005 mass%) is preferable. The indicator component of the royal jelly extract is 10-hydroxydecenoic acid.

  The extract from carrot (2) is preferably ginseng extract. Panax ginseng extract is a component obtained by extracting from Panax ginseng. Panax ginseng (CA Meyer) is a perennial plant belonging to the family Aralaceae, and is the base plant of the herbal medicine “Ginseng Radix”. Although appropriate parts, such as a flower, a stem, a leaf, and a root, can be used, it is preferable to use a root.

  Carrots may be subjected to extraction as they are, but considering extraction efficiency, it is preferable to perform extraction after processing such as shredding, drying, and pulverization. Extraction is performed by immersing in an extraction solvent. In order to increase the extraction efficiency, the mixture may be stirred, homogenized in an extraction solvent, or hydrolyzed by an enzyme treatment or the like. The extraction temperature is suitably about 5 ° C. to the boiling point of the extraction solvent. The extraction time varies depending on the type of extraction solvent and the extraction temperature, but is suitably about 4 hours to 14 days.

  Pressure control may be performed to increase extraction efficiency.

  Examples of the extraction solvent include water, lower alcohols such as methanol, ethanol, propanol, and isopropanol, polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, dipropylene glycol, and glycerin, diethyl ether, and dipropyl ether. Polar organic solvents such as ethers, esters such as ethyl acetate and butyl acetate, and ketones such as acetone and ethyl methyl ketone can be used. Further, physiological saline, phosphate buffer, phosphate buffered saline, or the like may be used.

  The blending amount of the extract from carrot with respect to the composition for external use of the skin of the present invention is 0.001 to 1% by mass (nonvolatile content 0.000013 to 0.013% by mass), particularly 0.01 to 0.1% by mass (nonvolatile content). 0.00013-0.0013 mass%). The index components of carrot extract are ginsenoside Rb1 and ginsenoside Rg1.

  Examples of (3) that can be used as a specific fat-soluble vitamin include vitamin A or fatty acid ester thereof, vitamin D, vitamin E, fatty acid ester of vitamin B2, fatty acid ester of vitamin B6, fatty acid ester of vitamin C, Pantothenol and niacin can be mentioned, and one or more of these can be used. It is particularly preferable to use at least a fatty acid ester of vitamin A and vitamin B2 or a fatty acid ester of vitamin B6, particularly at least vitamin A or a fatty acid ester thereof and a fatty acid ester of vitamin B6.

  As the fatty acid of the fatty acid ester of vitamin A, a saturated or unsaturated fatty acid having 8 to 26 carbon atoms is preferable, and a saturated or unsaturated fatty acid having 12 to 18 carbon atoms is particularly preferable. The fatty acid may be linear or branched, but is preferably linear. Saturated fatty acids are also preferred. An example is palmitic acid.

  The fatty acid of the fatty acid ester of vitamin B2 is preferably a saturated or unsaturated fatty acid having 2 to 26 carbon atoms, particularly preferably a saturated or unsaturated fatty acid having 2 to 12 carbon atoms. The fatty acid may be linear or branched, but is preferably linear. Saturated fatty acids are also preferred. An example is butyric acid. Generally it has a plurality of fatty acid ester groups, preferably 3-4.

  As the fatty acid of the fatty acid ester of vitamin B6, a saturated or unsaturated fatty acid having 8 to 26 carbon atoms is preferable, and a saturated or unsaturated fatty acid having 12 to 18 carbon atoms is particularly preferable. The fatty acid may be linear or branched, but is preferably linear. Saturated fatty acids are also preferred. For example, hexyl decanoic acid can be mentioned. In general, it has a plurality of fatty acid ester groups, preferably three.

  By using the specific fat-soluble vitamin, the action of suppressing the formation of wrinkles on the skin, and the action of maintaining and improving the firmness and elasticity of the skin become more prominent.

  In particular, the use of vitamins A enhances the metabolic function of the stratum corneum and enhances the action of excreting keratinized skin cells, and the effects of the present invention are more prominent. Further, by using vitamin B6, a moisturizing action by promoting filaggrin production is obtained, and the effects of the present invention are more remarkably exhibited. In order to increase skin permeability, vitamin B6 is used after being esterified with a fatty acid.

  The fat-soluble vitamin (3) is generally blended in the form of an emulsion, and in the present invention, the average particle size (lipophilic phase) of the emulsion is preferably 100 nm or less, particularly preferably 5 to 100 nm. In general, fat-soluble vitamins are micellized with a film agent (eg, a compound for forming micelles) in a solvent (generally a polyhydric alcohol or water). Further, the micellized emulsion may be added with an ion-sensitive matrix component such as alginic acid or encapsulated in microcapsules. By using the fat-soluble vitamin having such a particle size, the stability of the emulsion and the permeability to the skin are improved.

  As for the compounding quantity with respect to the lipophilic phase in the composition for external use of the skin of this invention of a fat-soluble vitamin, 0.001-1 mass% and 0.002-0.5 mass% are preferable.

  The external composition for skin of the present invention can contain the following components in addition to the components (1) to (3).

Minerals such as zinc, trace elements (eg, copper, manganese, iodine, molybdenum, selenium, chromium and cobalt);
Sugars (polysaccharides, trehalose, hyaluronic acid),
Protein (collagen, elastin),
Examples thereof include chondroitin, polyhydric alcohol (glycerin), pyrrolidone carboxylic acid (PCA), lactate, urea, ceramide, phospholipid, coenzyme Q10, α-lipoic acid and the like.

In addition, Yokuinin, Licorice, Kouca, Bowfu, Chinpi, Peonies, Psycho, Ginger, Artemisia,
Achacha, Aloe, Ireisen, Turmeric, Ages, Ogon, Oubak, Auren, Licorice, Keihi, Hawthorn, Salamander, Giant, Shikon, Senega, Senkyu, Taiso, Chimpi, Toki, Spruce, Carrot, Bacmondow, Honey, Bukuryo, Buttonpi , Mokutsu, Yokuinin,
Arnica, Aloe vera, Panax ginseng, Cascon, Cucumber, Chlorella, Atlantic squirrel, Atlantic white willow, Hypericum perforatum, Elderberry, Assembly, Chamomile, Zeni mallow And herbal medicines such as Kojisou), Sakuhakuhi, Hammamelis, Button, Maronnier, Rosemary and the like.

Furthermore, various amino acids;
Mention may be made of water-soluble vitamins such as thiamine (VB1), riboflavin (VB2), niacin (nicotinic acid) (VB3), pantothenic acid (VB5), pyridoxine (VB6), biotin (VH), ascorbic acid (VC) .

  Other fat-soluble vitamins include ergocalciphenol (VD), tocopherol, and tocotrienol (VE).

  Base ingredients include, for example, purified water, lower alcohols, polyhydric alcohols (eg, glycerin, sorbitol, polyethylene glycol, propylene glycol, butylene glycol), fats and oils, surfactants, ultraviolet absorbers, ultraviolet scattering agents, thickening agents. Agents, dyes, pigments, preservatives, fragrances and the like can be used. Glycerin is preferable, and this improves the stability of the emulsion and the permeability to the skin.

  Furthermore, an emulsifier, a fragrance | flavor, a coloring agent, antiseptic | preservative, etc. can be mix | blended.

  Examples of the colorant include safflower pigment, anthocyanin, turmeric extract, caramel, carotene, licorice extract, gardenia extract, kumazasa extract, saffron extract, zinc oxide, titanium oxide, sycon extract, copper chlorophyll, green tea extract, bengara, riboflavin, Examples include green tea powder and rose oil.

  Examples of preservatives include benzoic acid, sodium benzoate, ethanol, sodium edetate, BHA, BHT, phenoxyethanol, citric acid, sodium citrate, salicylic acid, sodium salicylate, sorbic acid, potassium sorbate, propylene glycol, l- Mentor, dl-camphor and eucalyptus oil.

  The external composition for skin of the present invention can be produced, for example, as follows. That is, the components (1) to (2) and the aqueous phase component of glycerin (and other components as appropriate) are heated to room temperature or, if necessary, 80 ° C. or lower, and mixed well to make uniform. . Similarly, the oil phase component containing (3) is also warmed to room temperature or, if necessary, 80 ° C. or less and mixed well to make it uniform. While stirring the aqueous phase component at room temperature or 80 ° C. or lower, the oil phase component containing (3) was gradually added and emulsified. This is stirred and cooled to obtain the external composition for skin of the present invention. This composition is generally an O / W emulsion composition.

  As described above, the fat-soluble vitamin (3), which is an oily component, is generally in the form of an emulsion, and the average particle size (lipophilic phase) of the emulsion is generally maintained after the emulsification. Therefore, the average particle size of the lipophilic phase of the obtained external O / W emulsion composition of the present invention is generally 100 nm or less.

  In order to deliver (administer) the active ingredient of the composition for external use of the skin of the present invention to a target site such as a damaged site of the skin, the composition is preferably in an emulsified state, and the average particle size thereof is in an emulsified state. In order to maintain the stability, it is preferably 0.1 to 500 μm and 1 to 200 μm. When the composition is an O / W emulsified composition as described above, it is general that the composition is further in an emulsified state as described above.

  In order to control the release amount of the active ingredient according to the amount absorbed from the skin, when the composition is porous microsphere or hollow microsphere, silicon dioxide, aluminum silicate, aluminum metasilicate, etc. are used as materials. It is preferable. In general, the average particle diameter is preferably 0.1 to 500 μm and 1 to 200 μm for the feeling of use and suspension stabilization.

  In the present invention, the production of type IV collagen can be promoted through the type IV collagen-producing ability of the extract from the blackwood azuki bean, and the laminin 5 production ability can be promoted through the ability to produce the laminin-5 contained in the extract from the black-tailed redwood. Production can be promoted. Thereby, reconstruction of the basement membrane structure can be induced, and skin aging symptoms can be prevented and improved. And such an effect | action is considered to have improved remarkably by presence of the other active ingredients (2)-(3) of the composition of this invention.

  The extract from black azuki bean is considered to have a matrix metalloprotease inhibitory action, which can inhibit the activity of matrix metalloprotease, and in particular, matrix metalloprotease-2 and / or matrix metalloprotease. -9 activity can be inhibited. Thereby, reconstruction of the basement membrane structure can be induced, and skin aging symptoms can be prevented and improved.

  The skin cosmetic containing the external composition for skin of the present invention is not particularly limited, and examples thereof include ointments, creams, emulsions, lotions, packs, foundations, lip balms, bath preparations, hair arts, hair lotions, soaps. And body shampoos. In particular, milky lotion, lotion, and cream are preferable because the form of the O / W emulsion can be utilized.

  Hereinafter, the present invention will be described in detail by way of examples. The present invention is not limited to the examples.

[Example 1]
[Laminin-5 production promoting effect test]
Human normal neonatal skin epidermal keratinocytes (NHEK) were cultured under conditions of 37 ° C. and 5% CO ₂ -95% air using growth factor-containing Epilife KG-2, and then recovered by trypsin treatment. The collected cells were diluted with BPE-free medium to a concentration of 1.0 × 10 ⁵ cells / mL, then seeded on a 24-well microplate at 500 μL per well, and cultured overnight. After completion of the culture, 500 μL of sample solution dissolved in BPE-free medium was added to each well and incubated for 24 hours. Then, 100 μL of the supernatant was transferred to an ELISA plate and adsorbed on the plate at 4 ° C overnight. . Thereafter, the solution was discarded and washed with a physiological phosphate buffer (PBS-T) containing 0.05% Tween-20. Thereafter, a blocking operation was performed with a phosphate physiological buffer containing 1% bovine serum albumin. The solution was discarded, washed with PBS-T, and reacted with anti-human laminin-5 antibody (mouse IgG, manufactured by Chemicon). The solution was discarded, washed with PBS-T, and reacted with biotin-labeled anti-mouse IgG (Amersham Biosciences). The solution was discarded, washed with PBS-T, reacted with streptavidin-peroxidase complex (manufactured by Calbiochem), and then the same washing operation was performed to perform a color reaction.

  Laminin-5 production promotion rate (laminin-5 production enhancing action) was calculated as (%) with the absorbance at the time of sample preparation solvent addition being 100%.

  As said sample solution, each density | concentration of the non-volatile content (3,10,30,100 microgram / mL) of a black-spotted azuki bean extract, each density | concentration of the non-volatile content (3,10,30,100 microgram / mL) of the ginseng extract The solution of was used. The results are shown in Table 1 and Table 2.

  Note) DMSO: Dimethyl sulfoxide

  From Table 1 above, it can be seen that both the black-and-white azuki bean extract and the carrot extract have the ability to promote laminin-5 production. Table 2 also shows that the effect is enhanced by the use of both.

[Example 2]
[Type IV collagen production promoting effect test]
Human normal skin fibroblasts (NB1RGB) were cultured in DMEM (Dulbecco's Modified Eagle Medium) containing 10% by weight fetal bovine serum (FBS) at 37 ° C and 5% CO ₂ -95% air. Thereafter, it was recovered by trypsin treatment. The collected cells were diluted to a concentration of 1.6 × 10 ⁵ cells / mL and then seeded at 100 μL per well in a 96-well microplate and cultured overnight. After completion of the culture, add 150 μL of the sample solution dissolved in DMEM containing 0.25% by mass fetal bovine serum (FBS) to each well, incubate for 72 hours, and then transfer 100 μL of the supernatant to an ELISA plate at 4 ° C. After adsorbing on the plate at night, the solution was discarded and washed with a phosphate buffered saline (PBS-T) containing 0.05% Tween-20. Thereafter, a blocking operation was performed with a physiological phosphate buffer containing 1% bovine serum albumin. The solution was discarded, washed with PBS-T, and reacted with anti-human collagen type IV antibody (rabbit IgG, manufactured by Novotech). The solution was discarded, washed with PBS-T, reacted with HRP-labeled anti-rabbit IgG antibody, and then the same washing operation was performed to perform a color reaction.

  The type IV collagen production promotion rate (%) was calculated by performing the above ELISA using a standard product, preparing a calibration curve, and assuming that the amount of type IV collagen produced when the sample preparation solvent was added was 100%.

  As the sample solution, a solution of each concentration of the non-volatile content (300, 600, 900 μg / mL) of the black bean extract is used, and a solution of each concentration of the non-volatile content (300, 600, 900 μg / mL) of the royal jelly extract is used. It was. The results are shown in Table 3 and Table 4.

  From Table 3 above, it can be seen that both the black and white extract and the royal jelly extract have the ability to promote type IV collagen production. Table 4 also shows that the effect is enhanced by the use of both of these.

[Example 3]
[Fibroblast proliferation promoting effect]
Human normal skin fibroblasts (NB1RGB) were cultured with DMEM containing 10% by weight fetal bovine serum (FBS) under conditions of 37 ° C and 5% CO ₂ -95% air, and then recovered by trypsin treatment. did. The collected cells were diluted to a concentration of 7.0 × 10 ⁴ cells / mL, then seeded at 100 μL per well in a 96-well microplate, and cultured overnight. After completion of the culture, 150 μL of a sample solution dissolved in DMEM containing 0.25 mass% fetal bovine serum (FBS) was added to each well and cultured for 72 hours. Cell proliferation activity was determined by measuring cell viability using the Alamar blue assay. Specifically, the medium was removed from each well, and 100 μL of Alamar blue dissolved in DMEM containing 5% by mass of fetal bovine serum (FBS) at a final concentration of 10% was added to each well. After culturing for 3 hours, the fluorescence intensity at 544 nm / 590 nm was measured. Then, from the obtained measurement results, the cell viability when the sample preparation solvent was added was calculated as 100%.

  As said sample solution, the solution of each concentration of the non-volatile content (300, 600, 900 μg / mL) of the royal jelly extract, and the solution of each concentration of the non-volatile content (600, 800, 1000, 1200, 1500 μg / mL) of the ginseng extract Was used. The results are shown in Table 5.

  From Table 5 above, it is recognized that both the royal jelly extract and the ginseng extract have the ability to promote fibroblast proliferation.

[Example 4]
Test cosmetics (samples A to D) having the composition shown in the following table were prepared as follows.

note)
BG: Butylene glycol

  By putting the lipophilic phase and the aqueous phase component prepared with vitamin A, vitamin B6 derivative and film agent shown in the above table to an average particle size of 50 to 100 nm into a stirring tank at room temperature and stirring with a disper for 30 minutes Mixing was performed to obtain Sample A (lotion).

  A thickener (acrylic acid / alkyl acrylate (C10-30)) crosspolymer (manufactured by Lubrizol Advanced Materials) was prepared in the same manner as sample A, except that it was previously dispersed to obtain sample B (essence). .

  Encapsulate the lipophilic phase prepared in vitamin A, vitamin B6 derivative and membrane agent shown in the above table in an average particle size of 50 to 100 nm in ion-sensitive microcapsules, and add the other oil phase and water phase components, The mixture was stirred and emulsified for 10 minutes to obtain Sample C (milky lotion).

  Encapsulate the lipophilic phase prepared in vitamin A, vitamin B6 derivative and membrane agent shown in the above table in an average particle size of 50 to 100 nm in ion-sensitive microcapsules, and add the other oil phase and water phase components, The mixture was stirred for 10 minutes and emulsified to obtain sample D (cream).

  Next, an evaluation test is performed on the skin lotion a prepared based on the sample A, the essence b prepared based on the sample B, the emulsion c prepared based on the sample C, and the cream d prepared based on the sample D. went.

  Forty-nine females (35 to 54 years old) with dry skin to normal skin who are worried about skin roughness, dryness, sagging, and fine wrinkles, 35 to 54 years old, actually used the above skin cosmetics and confirmed the effect. The subject was allowed to apply an appropriate amount of lotion a, essence b, milky lotion c, and cream d in this order to the skin after washing in the morning and night. This was continuously used for one month, and the skin condition was compared between the state before the start of the test and the one after the test cosmetic was used for one month. ”,“ Skin elasticity increased. ”,“ Skin gloss has improved. “The skin became brighter. ”,“ The wrinkles on the eyes are getting thinner. "" The texture has become finer. "I didn't feel any irritation." The evaluation items were evaluated.

  The results of the test are shown in the table below as the number of those who answered “yes” for each item among 49 subjects.

  Furthermore, the moisture content of the stratum corneum before the start of the test and after one month of use of the test cosmetics is measured with the Skicon-200 (manufactured by IBS). (Both made by Courage + Khazaka) were used as indicators of the degree of increase in moisture content, the degree of improvement in rough skin, and the degree of improvement in firmness, respectively. Regarding the above-described Cutometer MPA580, the height at which the skin was sucked and when it was released from the suction was measured and how fast the skin returned to its original height. That is, the skin is sucked for 2 seconds and opened for 2 seconds, and R7 (return rate in 0.1 seconds: height opened and returned in 0.1 seconds / height during 2 seconds suction) is used. Compared to before.

  Also, the condition of the skin texture was observed with a microscope, and when the photograph was taken, the fine texture condition was shown after use.

Claims (8)

As an active ingredient
(1) an extract from black-spotted azuki bean,
(2) Extract from royal jelly and / or carrot extract, and (3) Vitamin A or its fatty acid ester, Vitamin D, Vitamin E, Vitamin B2 fatty acid ester, Vitamin B6 fatty acid ester, Vitamin C fatty acid At least one fat-soluble vitamin selected from esters, pantothenol and niacin;
An external composition for skin, comprising:   (1) The composition for external use of the skin according to claim 1, wherein the extract from black-spotted azuki bean has type IV collagen production ability and laminin 5 production ability.   (2) The external composition for skin according to claim 1 or 2, wherein the extract from royal jelly and the extract from carrot have a fibroblast proliferation ability.   The components (1) and (2) are present in the hydrophilic phase (W phase), and the component (3) is present in the lipophilic phase (O phase). Skin external composition.   The composition for external use according to any one of claims 1 to 4, wherein the lipophilic phase (O phase) containing the component (3) has an average particle size of 100 nm or less.   The composition for external use on skin according to any one of claims 1 to 5, wherein the hydrophilic phase (W phase) contains glycerin.   The external composition for skin according to any one of claims 1 to 6, which does not contain parabens, synthetic pigments and synthetic fragrances.   An emulsion, lotion, cream or gel comprising the composition for external use of skin according to any one of claims 1 to 7.