JP2013091632A - Involucrin expression promoter - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an involucrin expression promoter capable of promoting expression of involucrin, a horny layer formation promoter, a humectant, a hair waving agent and a skin care external preparation.SOLUTION: In an involucrin expression promotor, a horny layer formation promoter, a humectant, a hair waving agent and a skin care external preparation, Veronica didyma var. lilacina or an extract therefrom is used as an active ingredient.

Description

  The present invention relates to an involucrin expression promoting agent, a stratum corneum formation promoting agent, a moisturizing agent, a hair waving agent, and a skin external preparation that promotes the expression of involucrin.

Involucrin is a protein produced in spinous cells according to the differentiation of epidermal keratinocytes, and is a strong insoluble membrane that lines the cell membrane of keratinocytes (cornified envelope; hereinafter also abbreviated as CE). ) Is one of the main components (Non-Patent Document 1). CE is formed by cross-linking and insolubilizing multiple CE precursor proteins including involucrin with the enzyme transglutaminase. Furthermore, involucrin that constitutes CE contributes to the formation of an intercellular lipid lamellar structure by covalently binding a lipid such as ceramide to a part of the involucrin (Non-patent Document 2). Such CE formation / maturation stabilizes the lamellar structure of the intercellular lipid, and the barrier function of the stratum corneum works normally, thereby enhancing the moisture retention function of the skin and the resistance to external stimuli.
Therefore, if involucrin expression is promoted in epidermal cells, the keratinization of epidermal cells is promoted, and the formation of a healthy stratum corneum is promoted, and the stratum corneum barrier function for protecting external stimuli and living bodies is healthy. As a result, it is considered that improvement of skin aging such as improvement of water retention ability of skin, prevention of pore conspicuousness, formation of wrinkles and reduction of texture pattern can be achieved.

  Recently, the present applicant has searched for the relationship between the hair shape and the gene expression of various genes in the hair root, and found that involcrine (IVL) in the hair root of a non-hairy person compared to the hair root part of the hairless person. It has been reported that the expression level of the gene is significantly increased, and it is possible to suppress or promote comb hair by controlling the expression of involucrin (Patent Document 1).

  Conventionally, examples of components that promote the expression of involucrin include Mesua Ferrea L. (patent document 2) of the Guttiferrae family, Craterellus cornucopioides (patent document 3), and the like. It has been reported that the CE formation effect can be expected through the action of promoting the expression of involucrin.

  On the other hand, it is known that Veronica didyma Tenore is a perennial plant belonging to the genus Sphagnum, which has analgesic and detoxifying effects. However, it promotes involcrine expression, promotes stratum corneum formation, and acts as a hair wave. It is not known that there is.

International Publication No. 2011/043330 Pamphlet JP 2005-213187 A JP 2005-89389 A

Steinert, PM. Et al., J. Biol. Chem., 270 (30), p17702-17711 (1995) Nemes, Z., Proc. Natl. Acad. Sci. USA, 96 (15), p8402-8407 (1999)

  The present invention relates to providing an involucrin expression promoting agent, a stratum corneum formation promoting agent, a hair waving agent, and a skin external preparation, which promote the expression of involucrin.

  The inventors of the present invention examined involucrin expression promotion. As a result, an extract of Inohokuguri, which is a plant of the genus Ligaceae, has an excellent involucrin expression promoting action, which promotes involucrin expression, promotes stratum corneum formation, and retains skin moisture It was found to be useful as a component or material for improvement of hair and hair waving.

That is, the present invention relates to the following 1) to 5).
1) An involucrin expression promoter containing inofugururi or an extract thereof as an active ingredient.
2) A stratum corneum formation promoting agent comprising inofuguri or an extract thereof as an active ingredient.
3) A moisturizing agent containing inofugururi or an extract thereof as an active ingredient.
4) A hair waving agent containing inuvenufu or an extract thereof as an active ingredient.
5) An external preparation for skin containing Inufuguri or an extract thereof.

  According to the present invention, a cosmetic, pharmaceutical or pharmaceutical product that has the ability to promote the production of involucrin, promotes cornification of horny cells, and promotes the formation of a healthy horny layer, thereby making the skin barrier function sound. Quasi-drugs, cosmetics for waving hair, quasi-drugs and the like can be provided.

The graph which shows the expression promotion effect of inboliclin.

In the present invention, “inufuguri” means inuvenu (Veronica didyma Tenore) belonging to the genus Sphagnumaceae.
Inufufu can be used in parts such as leaves, stems, buds, flowers, seeds, fruits, whole plants, etc., but whole plants are preferably used.

  Such Inufuguri can be used as it is or as a squeezed juice obtained by squeezing it, a dried product obtained by drying the plant itself, or a pulverized product thereof, or an extract extracted therefrom. preferable.

As the extract, various solvent extracts obtained by a known extraction method such as extraction of the above plant at room temperature or under heating, or extraction using an extractor such as a Soxhlet extractor, dilutions thereof, A concentrated solution or a dry powder thereof may be mentioned.
Known extraction methods include, for example, immersion, decoction, leaching, reflux extraction, supercritical extraction, ultrasonic extraction, and microwave extraction.

  As the extraction solvent used for obtaining the extract, either a polar solvent or a nonpolar solvent can be used. For example, water; alcohols such as methanol, ethanol, propanol and butanol; polyhydric alcohols such as propylene glycol and butylene glycol; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate; tetrahydrofuran and diethyl ether Linear and cyclic ethers such as polyethylene; polyethers such as polyethylene glycol; halogenated hydrocarbons such as dichloromethane, chloroform and carbon tetrachloride; hydrocarbons such as hexane, cyclohexane and petroleum ether; aroma such as benzene and toluene Group hydrocarbons; pyridines and the like, and these can be used alone or as a mixture. Among these, it is particularly preferable to use water, alcohol-based (alcohols and / or polyhydric alcohols (V / V)) solvents, and water-alcohol mixed solvents, and water-alcohol mixed solvents (V / V). Is preferred. Furthermore, alcohols are preferably alcohols, and among alcohols, methanol, ethanol, and propanol are more preferable, and ethanol is particularly preferable.

  For example, the inuvenir extract of the present invention can be obtained by using 1 to 50 parts by mass of an extraction solvent with respect to 1 part by mass of a plant and extracting at 4 to 100 ° C. for 0.5 to 30 days. More specifically, when water is used as the extraction solvent, it is preferably 5 to 30 parts by mass and 40 to 100 ° C. for 1 hour to 1 day with respect to 1 part by mass of the plant. Moreover, when using 40-60% ethanol aqueous solution as an extraction solvent, it is 3-20 mass parts with respect to 1 mass part of plants, Room temperature-under reflux, Preferably it is 20-50 degreeC, It is 1 hour-20 Days are preferred. Moreover, when using 75-100% ethanol aqueous solution, 3-20 mass parts with respect to 1 mass part of plants, 20 hours-under reflux, 1 hour-20 days are preferable.

  The above extract can be used as it is, but it can also be used by diluting, concentrating or lyophilizing the extract and preparing it in a powder or paste form. In addition to these extracted processed products, commercially available products may be used.

  The extract is preferably used after removing inactive impurities by a known technique such as liquid-liquid distribution, solid-liquid distribution, filtration membrane, activated carbon, absorption resin, ion exchange resin and the like. As the solvent used at this time, those exemplified for the extraction solvent may be used. Moreover, you may use these, after giving processes, such as a deodorizing and a decoloring, by a well-known method as needed. Moreover, when the extract is further purified, the known techniques and methods may be used.

As shown in the Examples below, the Inufuguri extract of the present invention promotes the expression of involucrin. If the expression of involucrin is promoted, stratum corneum formation is promoted and the skin barrier function can be made healthy. Thereby, improvement of the moisture retention ability of the skin (moisturizing effect) and improvement effect of pore conspicuousness are expected. Further, if expression of involucrin is promoted, it is considered that hair can be waved or waved can be promoted (see Patent Document 1).
Accordingly, inufufu or an extract thereof can serve as an involucrin expression promoter, horny layer formation promoter, moisturizer, and hair waving agent (hereinafter referred to as “involucrin expression promoter, etc.”), involucrin expression promoter, etc. Can be used to manufacture. That is, when applied to humans, it promotes the expression of involucrin or promotes stratum corneum formation, improves the skin barrier function, improves the moisture retention ability of the skin (moisturizing action), reduces wrinkles and reduces texture, etc. It can be used for prevention improvement, improvement of pore conspicuity, hair waving and the like.

  The involucrin expression promoters are cosmetics, pharmaceuticals and quasi drugs for promoting involucrin expression, promoting stratum corneum formation, improving skin moisture retention (moisturizing action), and waving hair. It may be a material or a preparation used in combination with the cosmetics, pharmaceuticals, quasi drugs and the like.

  “Wave” means that the straight hair is curled hair having curls, or the weak curled hair is made strong curled hair, and the curled shape is curly hair, curled hair, strong wave hair, etc. Any of these may be used.

  When the involucrin expression promoter of the present invention is used as cosmetics, quasi drugs or pharmaceuticals, it is in the form of an external preparation for skin, specifically, ointment, emulsified cosmetic, cream, emulsion, lotion, gel, It is preferably used in various forms such as aerosol. The external preparation for skin may be used on the surface of the skin after depilation and after removal of the stratum corneum, in addition to being used on the normal skin surface. When used for waving hair, it should be in the form of hair rinse, hair conditioner, hair treatment, hair lotion, hair pack, hair cream, conditioning mousse, hair mousse, hair spray, shampoo, leave-on treatment, etc. be able to. Such preparations are prepared according to general production methods, respectively, directly or pharmaceutically acceptable carriers such as various oils, surfactants, gelling agents, preservatives, antioxidants, solvents, alcohol, water, chelate. It can be obtained by mixing and dispersing together with an agent, a thickener, an ultraviolet absorber, an emulsion stabilizer, a pH adjuster, a dye, a fragrance and the like, and then processing into a desired form. In addition, for these cosmetics, quasi-drugs, pharmaceuticals, etc., the plant extracts, analgesic / anti-inflammatory agents, analgesics, bactericides, and astringents are appropriately selected according to the inuvenirs of the present invention or extracts thereof, and respective formulations. An emollient, a hormonal agent, vitamins, a moisturizer, an anti-inflammatory agent, a refreshing agent, an antiseborrheic agent and the like can be appropriately blended within a range that does not interfere with the effects of the present invention.

  In general, the content of the inuvenir of the present invention or the extract thereof in the cosmetic, quasi-drug or pharmaceutical product is preferably 0.0005 to 50% by mass as the solid content concentration, and 0.001 to 20%. It is more preferable to set it as the mass%.

There is no particular limitation on the dose of the inovenge of the present invention or the extract thereof in the above-mentioned pharmaceuticals and quasi-drugs as long as the effect is obtained, and it varies according to the condition, weight, sex, age or other factors of the subject. However, it is preferably 0.01 to 1500 mg, and more preferably 0.03 to 600 mg, for example, as Inufuguri or an extract thereof (in terms of dry matter) per adult (60 kg) per day. The preparation can be ingested / administered according to any ingestion / administration plan, but is preferably divided into once to several times a day and continuously administered for several weeks to several months.
In addition, the application target of the above-mentioned cosmetics, pharmaceuticals or quasi drugs is not particularly limited as long as it is necessary, but it promotes expression of involucrin, promotion of stratum corneum formation, sound skin barrier function, retention of skin moisture Humans for the purpose of improving the performance (moisturizing effect), improving the conspicuous pores and making the hair wavy are preferred.

  Hereinafter, the present invention will be described in more detail with reference to examples.

Production Example 1 Production of Inuvenufu Extract To 200 g dry product of Veronica didyma Tenore whole plant, Sichuan Province, China, 2000 mL of 50% ethanol was added, immersed in heating at 85 ° C. for 2 hours, and then filtered. To the residue after the filtrate, 2000 mL of 50% ethanol was added, and extraction was performed again at 85 ° C. for 1 hour. After filtration, the two filtrates were combined, concentrated under reduced pressure, and dried to obtain 13 g of Inufugururi extract. This was made into a 50% ethanol solution with a solid content of 1 w / v%, and used as an evaluation extract.

Example 1 Involucrin expression promoting action Involucrin ELISA Assay Kit (BT-650, BTI) was used for quantifying the protein content of involucrin.
A 24-well plate was seeded with HaCaT cells at 5 × 10 ⁴ cells / well and cultured. On the next day, the Inufufu extract obtained in Production Example 1 was replaced with a medium containing a final concentration of 0.5% v / v and further cultured for 24 hours. 24 hours after the addition of the extract, the cells were washed twice with PBS (Gibco), and 400 μl of a buffer containing Complete Mini Protease Inhibitor Cocktail (Roche) in 20 mM Tris-HCl (pH: 7.5) and 2 mM EDTA was added to the cells. did. One Protease Inhibitor Cocktail was added to 10 ml of buffer. Cells were detached and collected with a cell scraper in the presence of this buffer, and then sonicated with a sonicator. The supernatant collected by centrifugation of this solution was used as an lysate for evaluation.

  The total protein concentration of the lysate was quantified using BCA Protein Assay Kit (Thermo Scientific) according to the manufacturer's instructions. The lysate was diluted with buffer B (2 mM EDTA, 5 g / L Tween 20, 2.5 g / L Gelatin in PBS) to a total well of 3 µg per well of a 96-well plate and added to the 96-well plate Then, an anti-involucrin antibody (kit accessory, dilution ratio 1/300) was added and a primary antibody reaction was performed overnight at 4 ° C.

In addition, human involucrin protein (supplied with the kit) is added to another 96-well plate (Narugenunk) at a rate of 1 ng / well, and EDC (1-ethyl-3- (3-dimethylaminopropyl) carbodiimide, Kanto Chemical) 10 μg as a condensing agent Along with / well, antigen was adsorbed overnight at 4 ° C. On the next day, the solution of the plate on which the antigen was adsorbed was removed by aspiration, and 100 μl of 0.1M NH ₄ Cl was added and allowed to react at room temperature for 30 minutes. After the reaction, the plate was washed 4 times with distilled H ₂ O (Gibco) and once with buffer B, and 100 μl of the primary antibody reaction solution of the previous day was added and reacted at room temperature for 30 minutes. After this reaction, the plate was washed 5 times with buffer B, and the anti-rabbit-AP Conjugate antibody (dilution ratio 1/2000) attached to the kit was added and reacted at room temperature for 1 hour. After the reaction, 4 times with buffer B, then washed once with _{buffer D (1mM MgCl 2 .6H 2} O in 0.05M carbonate / bicarbonate ( Sigma), p-nitrophenylphosphate (1mg / ml in buffer D, Sigma) solution 100μl After the color development was confirmed, the absorbance at 405 nm was measured to determine the amount of involucrin protein.The expression level relative to the involucrin expression level in the control (no extract added) was 1.0. Values were determined and the results are shown in FIG.

  As is clear from FIG. 1, the Inufufu extract had an excellent inboliclin expression promoting effect.

Claims (5)

  An involucrin expression promoting agent comprising inuvenufu or an extract thereof as an active ingredient.   A stratum corneum formation promoter containing inuinufugu or an extract thereof as an active ingredient.   A moisturizing agent containing inofuguri or an extract thereof as an active ingredient.   A hair waving agent containing inunovuri or an extract thereof as an active ingredient.   An external preparation for skin containing Inufuguri or an extract thereof.

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