JP2003183121A - Composition for activating skin basal membrane - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a composition for activating the skin basal membrane which prevents, protects and improves the aging of the skin. <P>SOLUTION: This composition for activating the skin basal membrane contains a rice bran oil which accelerates the production of laminin 5 relating to promote the formation of the skin basal membrane and improvement of its functions. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a skin basement membrane-utilizing composition containing, as an active ingredient, rice bran oil that promotes the production of laminin-5, which is a constituent of skin basement membrane.

[0002]

2. Description of the Related Art Conventionally, it is known that aging, sun exposure, environmental stress, mental stress, etc. are involved in changes such as wrinkles, stains, dullness, and sagging that accompany aging of the skin. . Microscopic changes in the skin include reduction and denaturation of elastic fibers such as type I collagen, type III collagen and elastin in the dermis, and IV in the basement membrane.
Decrease and degeneration of type collagen and laminin and abnormal turnover of epidermal cells are occurring.

The basement membrane is a dense membrane composed of extracellular matrix proteins such as type IV collagen, laminin and fibronectin, and is present at the junction between the epidermis and the dermis in the skin. It is responsible for the connection between the epidermis and the dermis, is important for maintaining the structure of the skin, and is involved in the control of epidermal turnover and the transmission of information between the epidermis and the dermis.

Laminin 5 is known as one of the proteins having an especially important function among extracellular matrix proteins constituting the skin basement membrane. Laminin 5 is produced by epidermal cells and is involved in the binding between epidermal cells and basement membrane, as well as in binding to type VII collagen and also in binding between dermis and basement membrane (Rousselle, P.,
et al. J. Cell Biol. , 114, 5
67-576, 1991. , Rousselle,
P. , Et al. J. Cell Biol. , 13
8, 719-728, 1997. Chen, M .; , E
t al. J. Invest. Dermatol. ，
112, 177-183, 1999). Abnormality of the laminin 5 gene is a severe genetic disease characterized by exfoliation between the epidermis and the dermis and blistering, and combined epidermolysis bullosa (Herli).
tz's junctional epidermol
Laminin 5 is known to be an essential protein for the maintenance of normal skin structure because it causes ysis bullosa (Aberdam,
D. , Et al. , Nat. Genet. , 6,29
9-304, 1994. Pulkkinen, L .; ，
et al. , Genomics, 24, 357-36.
0, 1994. , Kiviriko, S .; , Et a
l. , Hum. Mol. Genet. , 4,959-9
62, 1995. ).

Recently, in exposed areas such as cheeks and eyelids, damage to the skin basement membrane such as tearing and multiplex of the basement membrane has been observed since the early 20s, and the deterioration has been remarkably deteriorated from the late 20s to the early 30s. Therefore, it was suggested that the damage of basement membrane is an important factor that causes skin aging. In addition, purified laminin-5 and substances that promote the production of laminin-5 promoted the formation of basement membrane and were shown to be useful substances for basement membrane care {J. Soc. Cosmet. Chem. Jpn. ，
Review, 35 (1) 1-7, 2001}.

From the above, a composition containing, as an active ingredient, purified laminin 5 having a function of promoting the formation of basement membrane and a plant extract or a compound that promotes the production of laminin 5 in order to prevent skin aging. Have been developed (JP-A-10-147515, JP-A-11-343226, JP-A-2
000-226308, Japanese Patent Application 2000-331318,
Japanese Patent Application No. 2001-151485).

[0007]

However, the composition containing purified laminin 5 as an active ingredient is stable, because laminin 5 is a very high molecular weight glycoprotein.
There is a problem with skin permeability, and a sufficient skin utilization effect cannot be expected. In addition, the substances known to date that promote the production of laminin 5 cannot be said to have sufficient stability and safety, and a sufficient skin utilization action cannot be expected. Therefore, it has been expected to develop a component having further excellent stability and safety.

[0008]

[Means for Solving the Problems] The present inventors have examined various components for their action of promoting the production of laminin 5 in epidermal cells. As a result, rice bran oil is laminin 5
The present invention has been completed by finding out that the production of Escherichia coli was promoted.

That is, the present invention is as follows. 1. A composition for stimulating a skin basement membrane containing rice bran oil 2. 2. A composition for promoting production of laminin 5, which contains rice bran oil. 1 for suppressing optical damage
Or the composition according to 2, 4. The composition according to 1, 2, or 3 which is for external use on the skin.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION The rice bran oil used in the present invention is
It is a semi-drying oil obtained from rice bran just after rice seed polishing and is commercially available. It is a pale yellow oily liquid that contains a large amount of free fatty acids, waxes, tocopherols, oryzanols, sterols, trace amounts of phospholipids, glycolipids, metals and the like in addition to neutral fats. The fatty acid components are mainly oleic acid, linoleic acid, and palmitic acid.

It is very stable against air and heat due to the effects of tocopherol and oryzanol. Almost no toxicity to skin cells such as epidermal cells and fibroblasts,
Very safe.

Oryzanol in rice bran oil has an effect of absorbing ultraviolet rays to protect the skin. It also has a cholesterol-lowering effect, and has a good texture and flavor, so it is used as an edible oil. Rice bran oil has a lot of oleic acid, and soap produced from rice bran oil is easily soluble in water and has excellent detergency, and is used as a household detergent. It is used as a natural sunscreen oil in Europe and America (Cosmetics Handbook, Nikko Chemicals Co., Ltd., Chuo Printing Co., Ltd.).

The effective amount of rice bran oil blended in the composition of the present invention is appropriately selected and determined according to the method of preparation of the rice bran oil, the form of the formulation, etc., but is not particularly limited to 0.0.
1-30% by weight is suitable.

The composition of the present invention containing rice bran oil promotes the production of laminin 5 in epidermal cells, is useful for activation and regeneration of the skin basement membrane, and has a function of the skin basement membrane reduced by the irradiation of ultraviolet rays. It enhances the binding force between basement membrane and basal cells and has a so-called photoaging regenerating effect.

The composition of the present invention includes oils and fats such as vegetable oils, higher fatty acids, higher alcohols, silicones, anionic surfactants, cationic surfactants, amphoteric surfactants,
Nonionic surfactants, preservatives, sugars, sequestering agents, polymers such as water-soluble polymers, thickeners, powder components,
An ultraviolet absorber, an ultraviolet blocker, a moisturizing agent such as hyaluronic acid, a fragrance, a pH adjusting agent, a desiccant and the like can be contained. Vitamins, skin stimulants, blood circulation promoters, indigenous bacteria control agents, active oxygen scavengers, anti-inflammatory agents, anti-cancer agents,
Other medicinal and physiologically active ingredients such as a whitening agent and a bactericidal agent may be contained.

Examples of oils and fats are camellia oil, evening primrose oil, macadamia nut oil, olive oil, rapeseed oil, corn oil, sesame oil, jojoba oil, germ oil, wheat germ oil,
Liquid oils and fats such as glycerin trioctanoate, cocoa butter,
Solid oils such as coconut oil, hardened coconut oil, palm oil, palm kernel oil, sorghum, sorghum kernel oil, hardened oil, and hardened castor oil, beeswax, candelilla wax, cotton wax, nuka wax,
Waxes such as lanolin, lanolin acetate, liquid lanolin, and sugar cane wax are mentioned.

Examples of hydrocarbons include liquid paraffin, squalene, squalane, and microcrystalline wax.

As the higher fatty acid, for example, lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, docosahexaenoic acid (D
HA), eicosapentaenoic acid (EPA) and the like.

Examples of higher alcohols include straight chain alcohols such as lauryl alcohol, stearyl alcohol, cetyl alcohol and cetostearyl alcohol, and branched chain alcohols such as monostearyl glycerin ether, lanolin alcohol, cholesterol, phytosterol and octyldodecanol. Can be mentioned.

Examples of silicones include chain polysiloxanes such as dimethylpolysiloxane and methylphenylpolysiloxane, and cyclic polysiloxanes such as decamethylcyclopentasiloxane.

Examples of anionic surfactants include fatty acid salts such as sodium laurate, higher alkyl sulfate ester salts such as sodium lauryl sulfate, alkyl ether sulfate ester salts such as POE lauryl triethanolamine lauryl sulfate, N-acyl sarcosinic acid, and the like. Sulfosuccinate,
Examples thereof include N-acyl amino acid salts.

Examples of the cationic surfactant include alkyl trimethyl ammonium salts such as stearyl trimethyl ammonium chloride, benzalkonium chloride and benzethonium chloride.

Examples of the amphoteric surfactant include betaine-based surfactants such as alkyl betaine and amidobetaine.

Examples of the nonionic surfactant include sorbitan fatty acid esters such as sorbitan monooleate and hydrogenated castor oil derivatives.

Examples of preservatives include methylparaben and ethylparaben.

Examples of the sequestering agent include edetate salts such as disodium ethylenediaminetetraacetate, edetic acid and sodium edetate.

Examples of the polymer include gum arabic, tragacanth gum, galactan, guar gum, carrageenan, pectin, agar, quince seed, dextran,
Pullulan carboxymethyl starch, collagen, casein, gelatin methyl cellulose, methyl hydroxypropyl cellulose, hydroxyethyl cellulose, sodium carboxymethyl cellulose (CMC), sodium alginate carboxy vinyl polymer (CARB0
POL) and other vinyl polymers.

Examples of the thickener include carrageenan, tragacanth gum, quince seed, casein, dextrin, gelatin, CMC, hydroxyethyl cellulose, hydroxypropyl cellulose carboxyvinyl polymer, guar gum, xanthan gum and bentonite.

As the powder component, talc, kaolin, mica, silica, zeolite, polyethylene powder, polystyrene powder, cellulose powder, inorganic white pigment, inorganic red pigment, titanium oxide coated mica, titanium oxide coated talc, colored titanium oxide coated. Examples thereof include pearl pigments such as mica, and organic pigments such as Red No. 201 and Red No. 202.

Examples of the ultraviolet absorber include para-aminobenzoic acid, phenyl salicylate, isopropyl para-methoxycinnamate, octyl para-methoxycinnamate, and 2,4-dihydroxybenzophenone.

Examples of the ultraviolet blocking agent include titanium oxide, talc, carmine, bentonite, kaolin, zinc oxide and the like.

As a moisturizer, polyethylene glycol,
Propylene glycol, dipropylene glycol, 1,
3-butylene glycol, 1,2-pentanediol,
Glycerin, diglycerin, polyglycerin, xylitol, maltitol, maltose, sorbitol, glucose, fructose, sodium chondroitin sulfate, sodium hyaluronate, sodium lactate, pyrrolidonecarboxylic acid, cyclodextrin and the like can be mentioned.

As the medicinal components, vitamin A oil, vitamins A such as retinol, and vitamin B ₂ such as riboflavin
S, _{B 6} such as pyridoxine hydrochloride, L- ascorbic acid, L- ascorbic acid phosphoric acid ester, L- ascorbic acid monopalmitate, L- ascorbic acid dipalmitate ester, L- ascorbic acid 2-glucoside C, pantothenic acid such as calcium pantothenate, vitamin D ₂ such as vitamin D ₂ and cholecalciferol; vitamin E such as α-tocopherol, tocopherol acetate, DL-α-tocopherol nicotinate Can be mentioned. Whitening agents such as placenta extract, glutathione, Yukinoshita extract, skin activating agents such as royal jelly, beech tree extract, blood circulation promoters such as capsaicin, zingerone, cantharisin tincture, ictamol, caffeine, tannic acid, γ-oryzanol, glycyrrhizic acid. Examples thereof include derivatives, glycyrrhetinic acid derivatives, anti-inflammatory agents such as azulene, amino acids such as arginine, serine, leucine and tryptophan, maltose sucrose condensates of resident bacterial control agents, lysozyme chloride and the like. Furthermore, chamomile extract, parsley extract, beech tree extract, wine yeast extract, grapefruit extract, honeysuckle extract, rice extract, grape extract, hop extract, rice bran extract, loquat extract, oyster extract, yokuinin extract,
Assembly extract, Merrilot extract, Birch extract, Licorice extract, Peony extract, Soapless extract,
Loofah extract, capsicum extract, lemon extract, gentian extract, perilla extract, aloe extract, rosemary extract, sage extract, thyme extract, tea extract, seaweed extract, cucumber extract, clove extract, carrot extract, horse chestnut extract, hamamelis extract, mulberry extract, etc. Can be mentioned various extracts.

The composition of the present invention is, for example, an aqueous solution, an oil solution,
Liquid agents such as emulsions and suspensions, semi-solid agents such as gels and creams,
It can be applied in the form of solid agents such as powder, granules, capsules, microcapsules, and solids. Prepared into these forms by conventionally known methods, lotions, emulsions, gels, creams, ointments, plasters, poultices, aerosols, suppositories, injections, powders, granules, tablets, pills. , Various forms such as syrup, troche, etc. These can be applied to the body by applying, sticking, spraying, drinking and the like. Especially among these dosage forms, lotion,
Emulsions, creams, ointments, plasters, poultices, aerosols and the like are suitable as external skin preparations. As cosmetics,
Skin lotion, milky lotion, cream, skin cosmetics such as packs, makeup base lotion, makeup cream, milky liquid or creamy or ointment type foundation, lipstick, eye color, cheek color makeup cosmetics, hand cream, Body creams such as leg creams and body lotions, bath salts, oral cosmetics,
It can be a hair cosmetic.

[0035]

The present invention will be further described with reference to examples, but the present invention is not limited to these examples.

Test results 1. Measurement of laminin-5 production promoting action in a three-dimensional model of human skin The three-dimensional model of human skin is TESTSKIN (LSE-
High) (Toyobo Co., Ltd.) was used for culturing according to the protocol. Fetal bovine serum (FBS) or rice bran oil (Tsukino Food Industry Co., Ltd.) was dissolved in vegetable squalane (Kao), and 80 μl was added on the tissue in the assay ring.
Incubated for hours. FBS is a known substance that promotes the production of laminin-5, and was used as a control here (Japanese Patent Laid-Open No. 1-58138)
1-343226). After washing several times with the culture medium to completely remove the drug, UVB irradiation was performed at 0 and 240 mJ / cm ² . A new drug was added and the cells were further cultured for 24 hours. Tissue is collected and tissue extraction solution {50 mM Tri
s-HCl (pH 7.5), 1% (Octylphe
noxy) polyethoxyethanol (Si
gma-Aldrich)} was added and homogenized. After removing the tissue piece by centrifugation at 15,000 rpm for 30 minutes, 50 mM Tris-HCl (pH 7.5)
It dialyzed at.

Using this tissue extract, ELISA (e
nzyme-linked immunosorben
The laminin-5 production promoting action was examined by t assay. The tissue extract was treated with 50 mM Tris-HCl (pH
It was diluted appropriately with 7.5) and adsorbed on a 96-well ELISA plate at 4 ° C. for 18 hours. After removing the cell culture supernatant, blocking solution {1% bovine serum albumin (B
It was immersed in PBS (−)} containing SA) and blocked at 37 ° C. for 1 hour. After washing with a washing solution {PBS (-) containing 0.05% polyoxyethylene (20) sorbitan monolaurate (Wako Pure Chemical Industries)}, the primary antibody solution {to the laminin γ2 chain prepared at 5 mg / ml with the washing solution Monoclonal antibody (clone D4B5) (Mizushi
ma, H., et al., Horm. Res., 50, 7-14,
1998.)} was added and reacted at 37 ° C. for 2 hours.
After washing, secondary antibody {biotinylated anti-mouse immunoglobulin G (Vector
laboratories)} was added and reacted at room temperature for 1 hour. After washing, enzyme solution {1 mg / ml with washing solution
Alkaline phosphatase avidin D (Ve
ctor laboratories)} was added and reacted at room temperature for 1 hour. After washing, substrate solution {1mg / ml
No p-nitrophenyl phosphate
0.75 M Tris-HCl (pH 10.3) containing (ICN Biomedicals, Inc.) was added, and after reacting at 37 ° C for 30 minutes, the absorbance at 405 nm was measured.

FBS 1.0%, rice bran oil 0.1%
%, 1.0% and 3.0% results are shown in FIG.
Shown in. Here, processing FBS at 1.0% means FB
It means that the stock solution of S is added to vegetable squalane at 1.0% for treatment. Also, rice bran oil 0.1%, 1.0%
And treatment with 3.0% means adding a stock solution of rice bran oil to vegetable squalane at 0.1%, 1.0% and 3.0% respectively. The drug was not treated, and UVB was not irradiated.
As a result, rice bran oil promoted laminin 5 production in a concentration-dependent manner both under UVB irradiation and under irradiation.
Incidentally, when FBS was treated, the production of laminin 5 was promoted as well.

Test results 2. In human skin three-dimensional model
Measurement of photodamage suppression effect The three-dimensional model of human skin is TESTSKIN (LSE-
high) and cultured according to the protocol
It was Dissolve FBS or rice bran oil in vegetable squalane
80 μl on the tissue in the assay ring,
Incubated for hours. Complete removal of chemicals by washing several times with culture solution
Then UVB 0 and 240 mJ / cm ^TwoIrradiate with
It was After adding a new drug and culturing for another 24 hours,
The culture solution was collected and centrifuged at 15,000 rpm for 30 minutes.
The tissue pieces were removed.

Using this tissue culture medium, the cytotoxicity was measured by measuring the lactate dehydrogenase (LDH) activity released from cells with damaged cell membranes. LDH
The activity was measured using an LDH-cytotoxicity test kit (Wako Pure Chemical Industries). A tissue culture solution appropriately diluted with PBS (-) was dispensed into a 96-well reaction plate,
The color developing solution was treated and allowed to react for 20 minutes at room temperature. After processing the reaction stop solution, 560n by microplate reader
The absorbance at m was measured to evaluate cytotoxicity.

FBS 1.0%, rice bran oil 0.1%
%, 1.0% and 3.0%, the results are shown in FIG.
Shown in. The drug was not treated and UVB was not irradiated, and the cytotoxicity was evaluated as 100%. In the case of non-irradiation with UVB, the cytotoxicity rate was almost the same regardless of the drug treatment. On the other hand, when UVB was irradiated, the cytotoxicity increased about twice as much as that in the untreated case, but when treated with rice bran oil, the cytotoxicity decreased in a concentration-dependent manner. When FBS was treated, the cytotoxicity rate also decreased.

The formulation examples of the present invention are shown below. Prescription example 1. Cream A cream was produced according to the following formulation (unit is mass%). (1) Stearyl alcohol 6.0% (2) Stearic acid 2.0% (3) Hydrogenated lanolin 4.0% (4) Squalane 9.0% (5) Octyldodecanol 10.0% (6) POE (25) Cetyl alcohol ether 3.0% (7) Glycerin monostearate 2.0% (8) Rice bran oil 1.0% (9) Preservative proper amount (10) Perfume proper amount (11) 1,3 butylene glycol 6 0.0% (12) PEG 1500 4.0% (13) Purified water Residual components (1) to (10) above are heated and dissolved at 80 ° C to form an oil phase. The components (11) to (13) are heated and dissolved at 70 ° C. to obtain an aqueous phase. The aqueous phase was gradually added to the oil phase to emulsify, and the mixture was cooled to 40 ° C with stirring and further cooled to 30 ° C with stirring to obtain a cream.

Prescription example 2. tablet A tablet was produced according to the following formulation (unit is mass%). (1) Rice bran oil 20.0% (2) Lactose 65.0% (3) Corn starch 14.0% (4) Guar gum 1.0%

[0044]

INDUSTRIAL APPLICABILITY As described above, the composition of the present invention containing rice bran oil promotes the production of laminin 5 in epidermal cells and promotes the maintenance of the structure and function of the skin basement membrane. Therefore, for aged skin, especially for skin damaged by ultraviolet rays, youthful skin without wrinkles, spots, dullness and sagging can be obtained by preventing, preventing or improving structural changes and functional decline of the skin basement membrane. The state of can be maintained.

[Brief description of drawings]

FIG. 1 is a diagram showing the expression level of laminin 5 when a human skin three-dimensional model is treated with FBS and rice bran oil.

FIG. 2 is a diagram showing suppression of photodamage when a human skin three-dimensional model is treated with FBS and rice bran oil.

Front page continuation (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61P 43/00 111 A61P 43/00 111

Claims (4)

[Claims] 1. A skin basement membrane stimulating composition containing rice bran oil. 2. A composition for promoting the production of laminin 5, which contains rice bran oil. 3. The composition according to claim 1, which is used for suppressing light damage. 4. The composition according to claim 1, which is for external use on the skin.