JP2009242310A - Involucrin production promoter, transglutaminase-1 production promoter, e-cadherin production promoter and skin care preparation for external use for making skin pore inconspicuous - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a novel agent that promotes production of involucrin, transglutaminase-1 and E-cadherin, and a skin care preparation for external use for making skin pores inconspicuous. <P>SOLUTION: The involucrin production promoter, the transglutaminase-1 production promoter and the E-cadherin production promoter comprise as an effective ingredient an extract of a seed of Annona squamosa. The skin care preparation for external use for making skin pores inconspicuous comprises as an effective ingredient an extract of a seed of Annona squamosa. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a novel production promoter for involucrin, transglutaminase-1 and E-cadherin, which is involved in normal stratum corneum formation, and a skin external preparation for reducing conspicuous pores.

  The conspicuousness of pores is always listed as the top of women's skin problems. The structure of the conspicuous pores is that the pores (funnels) are not widened, but the periphery of the pores is depressed like a mortar, and that part is conspicuous as a shadow. One of the causes of such a pore structure is the keratinization of horny layer cells around the pores. In prominent pores, the amount of sebum secretion increases, and the unsaturated fatty acids in sebum, as well as the unsaturated fatty acids produced by the degradation of triacylglycerol and acylglycerol in sebum by lipase, cause inflammation, resulting in keratin. It is known to cause oxidization (Non-patent Document 1).

It is known that the expression of related proteins / enzymes (involucrin, transglutaminase-1, etc.) is involved in the process of epidermal cells keratinizing and becoming stratum corneum cells. The stratum corneum is composed of a keratin fiber as a main component and a cornified envelope (keratinous thickened membrane, hereinafter abbreviated as CE) which is a membrane lining protein. CE is formed by cross-linking and insolubilizing a plurality of CE precursor proteins including involucrin produced in cells according to the keratinization process with the enzyme transglutaminase (Patent Document 1).
In recent years, it has been clarified that it is important for skin structure maintenance that cells of epidermis adhere to each other. There are various modes of cell adhesion, and one of them is known as an adhesion band, and cadherin is an adhesion molecule responsible for its function. The cadherin expressed on the cell membrane binds to the same kind of cadherin expressed in another cell in a calcium concentration-dependent manner, thereby intercellular adhesion. About 20 cadherins are known as a family, but the cadherin species present in the epidermis is E-cadherin. Since E-cadherin plays an important role in maintaining the structure of the skin epidermis, it is thought to be important for normal epidermal function.
Therefore, by promoting the production of these proteins and enzymes involved in the keratinization process and cell-cell adhesion, normal stratum corneum formation is promoted, and as a result, the conspicuous pores can be expected to be reduced.

On the other hand, since an external preparation for skin is directly applied to the skin, high safety is required for a medicinal agent used as an active ingredient. Therefore, conventionally, the medicinal effects of various plant extracts have been studied. For example, although a composition for the head comprising an extract derived from a banciaceae plant and a cosmetic composition containing steam distilled water of a banciaceae plant have been proposed (Patent Documents 2 and 3), The action on proteins and the like involved in the conversion process and cell-cell adhesion has not been known so far.
JP 2005-213187 A JP-A-6-40861 Japanese Patent Application Laid-Open No. 2001-220312 Fragrance Journal, March 2004, pages 41-47

  The present invention reduces the conspicuousness of pores by providing production promoters for involucrin, transglutaminase-1 and E-cadherin involved in normal stratum corneum formation, and by promoting normal stratum corneum formation It is an object to provide an external preparation for skin.

As a result of various studies to solve the above problems, the inventor of the present invention has an excellent production with respect to involucrin, transglutaminase-1, and E-cadherin, which are involved in normal stratum corneum formation, in the extract of Van Reishi seeds. It is found that even if it is added to the skin external preparation, the production promoting action for those proteins can be effectively exhibited, and further studies are made based on this finding to complete the present invention. It came to.
That is, the present invention provides an involucrin production promoter, a transglutaminase-1 production promoter, and an E-cadherin production promoter containing an extract of seeds of Vannashi (Annona squamosa) as an active ingredient in order to solve the above problems. To do.
From another point of view, according to the present invention, a skin external preparation for reducing pore conspicuousness containing an extract of van radish seeds as an active ingredient; and a skin external preparation for normalizing the stratum corneum containing an extract of van radish seeds as an active ingredient Is provided.

According to the present invention, it is possible to provide an involucrin production promoter, transglutaminase-1 production promoter, and E-cadherin production promoter with good safety.
In addition, according to the present invention, a novel topical skin preparation that promotes the production of involucrin, transglutaminase-1 and E-cadherin to enhance normal stratum corneum formation, and as a result, can reduce the conspicuousness of pores. An agent can be provided.

Hereinafter, the present invention will be described in detail. In the present specification, “to” means a range including numerical values before and after.
The present invention relates to a production promoter for involucrin, transglutaminase-1 and E-cadherin, which comprises an extract of seeds of Annona squamosa as an active ingredient. Banglei is a species of the genus Vanleci, a semi-deciduous shrub or small tree distributed in tropical America and the like. The fruit is edible. The extract used in the present invention is an extract of seeds in the fruit of Banglei.

  The extract of the bancii seed used in the present invention can be prepared by a general method. Although it does not specifically limit as an extraction solvent, For example, Water; Lower monohydric alcohols, such as methyl alcohol and ethyl alcohol; Liquid polyhydric alcohols, such as glycerol, propylene glycol, and 1, 3- butylene glycol; Ketones, such as acetone; Ethyl One or more of ethers such as ether; esters such as ethyl acetate; and the like can be used. It is preferable to use water or a mixed solution of water and a lower alcohol. The extraction can be carried out by immersing the seeds of banreishi in a solvent at room temperature or under heating for a predetermined time. Moreover, pre-treatment such as drying, chopping, pressing or fermentation can be performed on the seed before extraction.

After the preparation, the extract of the van radish seed can be used as it is as the various medicinal agents. Further, if desired, it can be left as it is for an appropriate period of time for aging and then used as the medicinal agent. If necessary, it can be used after being subjected to a purification treatment such as deodorization and decolorization by filtration or ion exchange resin within a range that does not affect the effect of the present invention. Further, a fraction having high activity can be taken out and used by using a separation means such as liquid chromatography.
Commercially available products may be used as the extract of the van radish seed, for example, “SMS ANTI-WRINKLE (manufactured by SILAB)”.

  The extract of the van radish seeds may be in any form such as liquid, paste or gel. The liquid extract containing the extraction solvent can also be used after it is solidified by drying under reduced pressure or freeze drying. It can also be dried by spray drying or the like and used as a powder.

  The extract of Van Reishi seeds has the effect of promoting the production of involucrin, transglutaminase-1, and E-cadherin. As described above, involucrin and transglutaminase-1 are proteins / enzymes involved in the process of epidermal cells keratinizing and becoming stratum corneum cells, and E-cadherin is involved in cell adhesion, particularly in the skin epidermis. It is an adhesion molecule that plays an important role in maintaining the structure. The extract of Van Reishi seeds has a production promoting action on all of them, and when applied to the skin, it promotes normal stratum corneum formation, resulting in conspicuous pores due to keratinization of stratum corneum cells. Can be reduced.

  The present invention also relates to a skin external preparation for reducing conspicuous pores, which contains an extract of van radish seeds as an active ingredient. The content of the extract of the Van Reishi seeds in the external preparation for skin of the present invention is preferably 0.0001 to 20% by mass (hereinafter simply referred to as “%”) as a solid content, and more preferably 0.001 to 10%. If it exists in this range, the said extract can be mix | blended stably and the high reduction effect with respect to conspicuous pores can be exhibited.

  The external preparation for skin of the present invention can be prepared by blending the extract of the banley seeds with various forms of bases according to a conventional method. Furthermore, by blending the extract in combination with one or more other medicinal agents, it is possible to prepare a topical skin preparation that further enhances the medicinal properties or exhibits other medicinal properties along with the medicinal properties. Examples of other medicinal agents include whitening agents, UV protection agents, antibacterial agents, bactericides, sebum secretion regulators, anti-inflammatory agents, cell activators, active oxygen scavengers, and moisturizers. It is not limited to.

  Examples of whitening agents include ascorbic acid or derivatives thereof, arbutin, ellagic acid, linoleic acid, vitamin E and derivatives thereof, glycyrrhizic acid and derivatives thereof, tranexamic acid, placenta extract, chamomile extract, seaweed extract, coconut extract , Gokahi extract, Rice bran extract, Wheat germ extract, Saisin extract, Hawthorn extract, Sunpens extract, Shirayuri extract, Peonies extract, Sempukuka extract, Soy extract, Tea extract, Molasses extract , Beechlen extract, grape extract, micaika extract, mokka extract, saxifrage extract and the like.

Examples of the UV protection agent include paramethoxycinnamate-2-ethylhexyl, 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxybenzophenone-5 sodium sulfate, 4-t-butyl-4′- Methoxydibenzoylmethane, 2
-Phenyl-benzimidazole-5-sulfuric acid, titanium oxide, zinc oxide and the like.

  Examples of the antibacterial agent include benzoic acid, sodium benzoate, paraoxybenzoic acid ester, phenoxyethanol, and the like.

  Examples of the disinfectant include salicylic acid, benzalkonium chloride, isopropylmethylphenol, hinokitiol, sulfur and derivatives thereof, eucalyptus extract, clove extract, and dokudami extract.

  Examples of the sebum secretion regulator include, for example, Citrus extract, Licorice extract, Age extract, Ogon extract, Oat extract, Kudin extract, Hops extract, Watercress extract, Pyridoxine and its derivatives, Auren extract, An example of the extract is an extract of pearl millet, mint extract, mugwort extract and the like.

  Anti-inflammatory agents include, for example, glycyrrhizic acid and its derivatives, glycyrrhetinic acid and its derivatives, artea extract, ashitaba extract, arnica extract, ginseng extract, nettle extract, buckwheat extract, hypericum extract, chamomile extract Products, goldfish extract, comfrey extract, salvia extract, sicon extract, perilla extract, birch extract, gentian extract and the like.

  Examples of cell activators include caffeine, chicken crown extract, shell extract, shell extract, royal jelly, silk protein and its degradation products or derivatives thereof, lactoferrin or degradation products thereof, chondroitin sulfate, hyaluronic acid, etc. Mucopolysaccharides or their salts, collagen, yeast extract, lactic acid bacteria extract, bifidobacteria extract, fermentation metabolic extract, ginkgo biloba extract, barley extract, assembly extract, peanut extract, carrot extract, rosemary Extracts, glycolic acid, citric acid, lactic acid, malic acid, tartaric acid, succinic acid and the like are included.

  The active oxygen scavenger has an action such as suppression of lipid peroxide production. For example, superoxide dismutase, mannitol, quercetin, catechin and derivatives thereof, rutin and derivatives thereof, button pi extract, yashajitsu extract, melissa extract , Rakan fruit extract, retinol and its derivatives, vitamin A such as carotenoid, thiamine and its derivative, riboflavin and its derivative, vitamin B such as nicotinic acid and its derivative, vitamin E such as tocopherol and its derivative, dibutyl Examples include hydroxytoluene and butylhydroxyanisole.

  Examples of the humectant include proteins such as elastin and keratin or derivatives thereof, hydrolysates and salts thereof, amino acids such as glycine, serine, aspartic acid, glutamic acid, arginine and theanine and derivatives thereof, sorbitol, erythritol, Trehalose, inositol, glucose, sucrose and derivatives thereof, dextrin and derivatives thereof, saccharides such as honey, D-panthenol and derivatives thereof, urea, phospholipid, ceramide, shobu extract, diautum extract, senkyu extract, mallow extract Products, dokudami extract, hamamelis extract, bodaige extract, maronier extract, quince extract and the like.

  Moreover, arbitrary ingredients other than the medicinal agent of this invention can be mix | blended with the skin external preparation of this invention. Examples of such components include, but are not limited to, physiologically active substances such as amino acids, lipids, sugars, hormones, enzymes, and nucleic acids. In addition, components that are usually used in the production of cosmetics, quasi-drugs, skin external preparations and the like, for example, water (purified water, hot spring water, deep water, etc.), oil agents, Surfactant, metal soap, gelling agent, powder, alcohol, water-soluble polymer, film-forming agent, resin, inclusion compound, fragrance, deodorant, salt, pH adjuster, refreshing agent, plant / animal -Extracts derived from microorganisms, blood circulation promoters, astringents, antiseborrheic agents, chelating agents, keratolytic agents, enzymes, hormones, other vitamins and the like can be used as necessary.

  The external preparation for skin of the present invention has various forms such as powders such as powder and powder foundation; solids such as soap and lipstick; emulsions such as cream, emulsion and cream foundation; It is preferable that it is a cosmetic composition. However, it is not limited to these.

EXAMPLES Hereinafter, the present invention will be described more specifically with reference to examples. However, the scope of the present invention is not limited to the following examples.
[Example 1: Preparation of extract of Van Reishi seeds]
After pulverizing 100 galvanic seeds, 1.8 L of purified water was added and extraction was performed at room temperature for 7 days, followed by filtration to obtain 1.2 liters of baneraishi seed extract (dried solid content 4 in the extract). 0%).

[Example 2: Test for promoting production of involucrin and transglutaminase-1]
Human-derived keratinocytes were seeded and placed in a 5% CO ₂ incubator. Three days later, the bancii seed extract prepared in Example 1 was added at a concentration of 0.5 (v / v)%, and was allowed to stand again in the incubator. After 3 days, the cells were collected and RNA was extracted. The amount of mRNA for involucrin and transglutaminase-1 was determined by RNA quantitative PCR. The results are shown in the following table. In addition, in the following table | surface, the production amount of involucrin and transglutaminase-1 of the sample group without added vanilla seed extract was defined as 100%, and the ratio was shown as a percentage.

[Example 3: E-cadherin production promoting effect test]
1 × 10 ⁶ human-derived keratinocytes were seeded in a 10 cm petri dish and allowed to stand in a 5% CO ₂ incubator. Three days later, the extract of the banley seed prepared in Example 1 was added at a concentration of 0.5 (v / v)%, and left again in the incubator. After 3 days, the cells were collected to obtain a crude protein extract. The obtained crude protein extract was supplied to an 8% polyacrylamide gel and subjected to electrophoresis, and then the amount of E-cadherin protein produced was determined from the band detected by Western blotting. The results are shown in the following table. In the table below, the production amount of E-cadherin in the sample group without added vanilla seed extract was defined as 100% and shown as a ratio to that.

  From the results shown in the above table, it can be understood that the extract of Van Reishi seeds has a high production promoting action on involucrin, transglutaminase-1, and E-cadherin. Therefore, it can be expected that the extract of van radish seeds promotes the production of these proteins and the like and contributes to normal stratum corneum formation when applied to the skin.

[Example 4: Evaluation of pore conspicuous reduction effect]
(Preparation of lotion)
A lotion having the following composition was prepared by the following method.
(Prescription) (%)
(1) Glycerin 5.0
(2) 1,3-butylene glycol 6.5
(3) Polyoxyethylene (20E.O.)
Sorbitan monolaurate ester 1.2
(4) Ethyl alcohol 8.0
(5) Banglei extract ^{* 1} 4.0
(6) Preservative appropriate amount (7) Purified water remaining amount * 1 Prepared in Example 1

(Manufacturing method)
A. (3), (4) and (6) are mixed and dissolved.
B. (1), (2), (5) and (7) are mixed and dissolved.
C. A and B were mixed and uniformed to obtain a skin lotion.

  In addition, a lotion for comparative example was prepared in the same manner except that the extract of Banglei seed was not mixed.

(Evaluation of lotion)
A makeup for a comparative example in which 7 panelists who are conspicuous about pores have a skin lotion containing the above-prepared extract of vanilla seeds on one of the left and right cheeks, and a vanillai seed extract on the other side Water was applied twice a day for 4 weeks. The degree of conspicuous pores in the left and right cheeks was determined by the following four-stage visual evaluation, and the average of all the panels was determined.
(Evaluation criteria)
1: Pore is not conspicuous 2: Pore is conspicuous 3: Pore is conspicuous 4: Pore is conspicuous
(Changed to 4-level evaluation)

(Evaluation results)
The average value of the lotion of the Example which mix | blended the extract of the Van Reishi seed was 3.375 points before use, and was 2.375 points after use. On the other hand, the average value of the skin lotion of the comparative example was 3.375 points before use and 3.25 points after use. From this result, the extract of Van Reishi seed does not lose its efficacy even if it is blended with skin external preparations such as lotion, and contributes to normal stratum corneum formation by being applied to the skin. It can be understood that the conspicuous effect is reduced.

[Example 5: Latex] (Prescription) (%)
(1) Polyoxyethylene (10E.O.) sorbitan monostearate 1.0
(2) Polyoxyethylene (60E.O.) sorbite tetraoleate 0.5
(3) Glyceryl monostearate 1.0
(4) Stearic acid 0.5
(5) Behenyl alcohol 0.5
(6) Squalane 5.0
(7) Preservative 0.1
(8) Carboxyvinyl polymer 0.1
(9) Sodium hydroxide 0.05
(10) Ethyl alcohol 5.0
(11) Purified water Residual amount (12) Bunglei seed extract * 1 1.0
(13) Dipotassium glycyrrhizinate * 2 0.1
(14) Kujin extract * 3 0.2
(15) Citrus extract * 4 1.0
(16) Fragrance Appropriate amount * 1 Prepared in Example 1 * 2 Made by Maruzen Pharmaceutical * 3 Made by Maruzen Pharmaceutical * 4 Made by SILAB

(Manufacturing method)
A. Ingredients (1) to (7) are heated and mixed and maintained at 70 ° C.
B. A part of components (9) and (11) and (13) are heated and mixed and maintained at 70 ° C.
C. A is added to B, mixed and uniformly emulsified.
D. After cooling C, (12), (14), (15) and (8), (10), (16) dissolved in the remainder of (11) were added and mixed uniformly to obtain an emulsion.

[Example 6: Cream]
(Prescription) (%)
(1) Beeswaw 6.0
(2) Cetanol 5.0
(3) Reduced lanolin 5.0
(4) Squalane 30.0
(5) Lipophilic glyceryl monostearate 4.0
(6) Polyoxyethylene sorbitan monolaurate (20E.O) 2.0
(7) Banglei seed extract * 1 0.5
(8) Ages extract * 2 0.5
(9) Hop extract * 3 0.5
(10) mint extract * 4 0.5
(11) Preservative Appropriate amount (12) Fragrance Appropriate amount (13) Purified water Remaining amount * 1 Prepared in Reference Example 1 * 2 Made by Maruzen Pharmaceutical * 3 Made by Maruzen Pharmaceutical * 4 Made by Maruzen Pharmaceutical (production method)
A. Ingredients (1)-(6) and (11) are mixed and heated to keep at 70 ° C.
B. A portion of component (13) is heated and maintained at 70 ° C.
C. B was added to A, (7) to (10) and (12) dissolved in the remainder of (13) were mixed, and then cooled to obtain a cream.

[Example 7: Gel ointment]
(Prescription) (%)
(1) Carboxyvinyl polymer 1.0
(2) Triethanolamine 1.0
(3) 1,3 butylene glycol 10.0
(4) Banglei seed extract * 1 2.0
(5) Pyridoxine hydrochloride * 2 0.1
(6) Hop extract * 3 0.3
(7) Artemisia extract * 4 0.3
(8) Licorice extract * 5 0.3
(9) Watercress extract * 6 0.3
(10) Purified water remaining * 1 Manufactured in Reference Example * 2 Sigma Corporation * 3 Maruzen Pharmaceutical * 4 Maruzen Pharmaceutical * 5 Maruzen Pharmaceutical * 6 Maruzen Pharmaceutical

(Manufacturing method)
A. Components (1) and (3) to (10) are mixed and dissolved.
B. Ingredient (2) was added to A and mixed to obtain a gel ointment.

  Examples 5 to 7 were external preparations for skin that alleviate the conspicuousness of pores by being continuously applied to the skin.

Claims (4)

An involucrin production promoter containing an extract of the seeds of Van Reishi (Annona squamosa) as an active ingredient. A transglutaminase-1 production promoter comprising, as an active ingredient, an extract of a seed of Annona squamosa. An E-cadherin production promoter comprising an extract of seeds of Annona squamosa as an active ingredient. A skin external preparation for reducing the appearance of pores, containing an extract of seeds of Annona squamosa as an active ingredient.