JP2009084212A - Involucrin production promoter - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an external preparation for the skin, suppressing various kinds of irritations to the skin by improving skin barrier functions by promoting the involucrin production by epidermal cells. <P>SOLUTION: The external preparation for the skin, suppressing the various kinds of the irritations to the skin by improving the skin barrier functions comprises an extract of Leontopodium alpinum having effects for promoting the involucrin production. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

The present invention relates to a skin barrier function improving agent characterized by containing an extract of plant edelweiss as an involucrin production promoter. Examples of the field of use thereof include cosmetics, quasi-drugs, and external medicines. It can be used for external preparations such as skin. More specifically, it is an external preparation for skin characterized by containing an extract of plant Edelweiss as an involucrin production promoter. The present invention relates to a topical skin preparation that promotes keratinocyte involucrin production to promote keratinization of epidermal cells, and promotes formation of the stratum corneum, thereby improving the skin barrier function.

Conventional technology

Epidermal cells undergo differentiation induction until they are newly born and detach, synthesize various keratin molecular species, and express proteins necessary for keratinization. In particular, during the keratinization process of epidermal cells, proteins such as involucrin and loricrin appear from the spinous layer to the granular layer, and are cross-linked by enzymes such as transglutaminase to form a peripheral zone (cornified envelope). It forms a tough structure that lines the cell membrane. Furthermore, it has been suggested that it forms the basis of the epidermal barrier function together with intercellular lipids such as ceramide present between corneocytes.

The peripheral zone is formed by cross-linking many molecules such as involucrin, loricrin, SPR, elafin, cystatin A, S-100 protein family, desmosomal proteins in keratinocytes with sulfhydryl oxidase and transglutaminase. The peripheral zone is extremely important for the physical, chemical and biological barrier functions of the epidermis, and it is known that abnormalities in the peripheral zone due to the deficiency of the cross-linking enzyme cause severe keratoses.

On the other hand, in recent years, an increasing number of people complain of modulation on the skin due to stress, environmental pollution, and the like. In such human skin, the barrier function of the skin is abnormal, so it is easy to feel allergens such as chemical substances, mites, dust, etc., UV rays, etc., skin irritation, itching, rough skin, inflammation, etc. Or in pathological cases, it leads to diseases such as atopic dermatitis and contact dermatitis. Until now, various approaches have been made to improve sensitive skin. For example, in order to reduce irritation to the skin, use a topical skin preparation that does not contain as much as possible preservatives, alcohol, and other substances that can irritate the skin, and use many moisturizing ingredients such as collagen, elastin, and hyaluronic acid. The skin external preparation contained or the skin external preparation which mix | blended various anti-inflammatory agents has been used.
JP 10-182332 A JP 2000-212203 A Shimizu, Hiroshi, New Dermatology, 3-9, 2005.

However, these external preparations for skin temporarily supplement moisture on the surface of the stratum corneum, and do not exhibit sufficient effects only by suppressing inflammation, and their improvement has been desired.

Therefore, as a result of intensive studies to solve such problems, the present inventors have found that an extract of plant Edelweiss promoted the production of involcrine, a precursor protein of the peripheral zone (cornified envelope), and the stratum corneum cells The inventors have found that it has a function of promoting the improvement of the barrier function of the stratum corneum by promoting the keratinization of the corneal layer and completed the present invention.

That is, the present invention is an external preparation for skin characterized by containing an extract of plant Edelweiss as an involucrin production promoter, by promoting cornification of horny layer cells and improving the barrier function of the horny layer. Further, the present invention provides an external preparation for skin which can be expected to have a rough skin prevention effect, a stimulation alleviation effect and the like.

The edelweiss (Leontopodium alpinum) used in the present invention is an alpine plant belonging to the genus Leontopodium, and is also referred to as Hanausyukisou. In German, Edel means "noble" and Weiss means "white". The height will be 5-20cm and will bloom from June to September.

The part of the plant body in the plant extract used in the present invention is one place from a flower, a flower spike, a fruit skin, a fruit, a stem, a leaf, a branch, a branch and a leaf, a stem, a bark, a rhizome, a root bark, a root, a seed, or a whole plant. The above can be used. In addition to the extract obtained by directly extracting from these various extraction sites using a solvent, the extract may be extracted by adding a solvent to the squeezed liquid and / or residue obtained after the squeezing treatment.

The extraction solvent for obtaining the plant extract used in the present invention may be selected in consideration of the intended purpose and type of the product to be provided, or the processing performed later, for example, water; methyl alcohol, ethyl Lower monohydric alcohols such as alcohol; Liquid polyhydric alcohols such as glycerin, propylene glycol, and 1,3-butylene glycol; Ketones such as acetone and methyl ethyl ketone; Alkyl esters such as ethyl acetate; Hydrocarbons such as benzene and hexane; Diethyl ether It can be extracted and purified by using one or more of halogenated alkanes such as dichloromethane and chloroform.

The plant to be extracted is obtained by collecting the use site, pulverizing after drying, 1 to 1000 times by weight, particularly 10 to 100 times by weight of solvent, and in the case of room temperature extraction, 0 ° C. or more, especially It is preferably performed at 20 ° C. to 40 ° C. for 1 hour or longer, particularly 3 to 7 days. Moreover, you may heat-extract at 60-100 degreeC for 1 hour.

Each of the above-mentioned extracts obtained under the above conditions may be used as an extracted solution. However, if necessary, use a product that has been refined, filtered, concentrated, and powdered as necessary. I can do it.

The form of the plant extract used in the present invention may be any form such as liquid, solid, powder, paste, gel, etc., and an optimum shape is arbitrarily selected for constituting the final product. be able to.

The dosage form of the external preparation for skin of the present invention is arbitrary, and is capsule, powder, granule, pill, tablet, solid, liquid, gel, foam, emulsion, cream, ointment, sheet The shape can be in the form of an aerosol or aerosol. Furthermore, it can mix | blend and use for pharmaceuticals, quasi-drugs, cosmetics, or food-drinks. In particular, it is applied to external preparation compositions such as pharmaceuticals, quasi drugs, and cosmetic compositions applied to the outer skin.

Specific use forms of the present invention include aqueous components, oily components, plant extracts, animal extracts, powders, excipients, surfactants, oils, alcohols, pH adjusters, preservatives, antioxidants, A lotion, emulsion, cream, pack, powder, spray, ointment, dispersion, cleaning agent for external preparation composition by mixing thickeners, sweeteners, pigments, fragrances and the like as necessary. In addition, various dosage forms such as liquid, paste, capsule, powder, and tablet can be used.

The blending amount of the involucrin production promoter in the external preparation for skin of the present invention is slightly different depending on the expected degree of action and is not particularly limited, but is usually 0.0001% by mass or more in terms of solid content in the total amount of the preparation, preferably It is effective to make the concentration range 0.01 to 20.0 mass%.

Hereinafter, test examples relating to the effects of the involucrin production promoter according to the present invention will be shown and prescription examples applied to external preparations using the materials will be described, but it goes without saying that the examples are not limited thereto. .

[Preparation of plant extract]
50 ml of purified water was added to 1 g of the edelweiss whole plant dried and pulverized, and the mixture was extracted by heating at 80 ° C. for 1 hour. The extract was filtered and the residue dried under reduced pressure at 40 ° C. was dried. The dried product was prepared to be a 1% aqueous solution and used as a sample solution.

[Cell culture]
Cell: NHEK-Neo-Epidermal Kera (CAMBREX)
Medium: Epilife KG2 (Ca concentration: 0.06 mM)
D-MEM (Ca concentration: 1.8 mM)
NHEK-Neo-Epidermal Kera (CAMBREX), a normal human epidermal cell, was cultured in Epilife KG2 (Kurabo) medium.
The cells were seeded and cultured on a 50 cm ² plate at about 50% confluence. The next day various extracts were added. 48 hours after the addition, the cells were collected and used as Western blotting samples.

[Western blotting]
After electrophoresis on 7.6% acrylamide gel, transfer to PVDF membrane, SANTA CRUZ BIOTECHNOLOGY as involucrin antibody, INC Involucrin (H-120) sc-28557,
Human (rabbit
polyclonal), ECL ^TM Anti-rabbit IgG as secondary antibody,
Horseradish
Peroxidase linked whole antibody (from donkey) NA934V
(Amersham
Western blotting was performed using Bioscience UK Limited to confirm the amount of involucrin protein having a molecular weight of 68 KDa.

[Measurement of involucrin protein content]
The band intensity obtained by Western blotting was analyzed by Dolphin-DOC (WEALTEC). The area of the band intensity of the involucrin protein in the sample-free group was set to 100, and the band intensity area of the involucrin in each of the extract-added sections was proportionally calculated to obtain the amount of involucrin protein.

The experimental results are shown in Table 1. As a result of analysis of the band intensity area, when the control group was set to 100, indextrin production was 132 in the 560 ppm addition group and 122 in the 280 ppm addition group, and the involcrine production promoting effect by the edelweiss extract was confirmed. .

[Skin barrier function test]
A 2.5% SDS aqueous solution was applied to the inner skin of the human upper arm for 5 minutes a day, and this was repeated for 10 days to obtain a skin model in which the barrier function was destroyed. Then, the Edelweiss extract (2% and 5%) shown in the Example of Table 2 was added to the emulsion composition 100 shown in the paragraph number 0025 excluding the preservative and the antioxidant, and Example 1 and Example 2 were added. An emulsion of comparative product 1 was prepared. After 5 selected skin panelists applied each test product for 2 weeks, the skin water transpiration was measured with TEWA meter TM-210 (Courage + Khazaka). The results are shown in Table 2. From Table 2, the barrier function of the skin was destroyed by applying SDS, and the TEWL value increased to 38. On the other hand, by applying the additive according to the present invention, the TEWL value decreased to 19 in the composition containing 2% of the Edelweiss extract and decreased to 17 in the composition containing 5%. Here, from the fact that the TEWL value was 31 when the emulsion composition not added with the Edelweiss extract was applied, the group applied with the emulsion composition added with the Edelweiss extract clearly improved the skin barrier function. Was recognized.

[Emulsion composition]
(Ingredient name)
(weight%)
a) Beeswax
0.5
b) Petrolatum
2.0
c) Squalane 8.0
d) Sorbitan sesquioleate
0.8
e) Polyoxyethylene oleyl ether (20E.O.)
1.2
f) Preservatives and antioxidants
g) Purified water
The rest
h) 1,3-butylene glycol
7.0
i) Carboxyvinyl polymer
0.2
j) Potassium hydroxide 0.1
k) ethanol
7.0
Manufacturing method: Heat up to a) to f) and keep at 80 ° C. G) to i) are dissolved by heating, kept at 80 ° C., added to a) to f) and emulsified, and j) previously dissolved in part of g) is added and mixed uniformly. Then add k) at 50 ° C. and cool to 40 ° C. with stirring.

[Manufacture of various compositions]
Various compositions according to the present invention were prepared. Although the formulation example is shown below, this invention is not necessarily limited to these.

(1) Cream composition
(weight%)
a) Beeswax
2.0
b) Stearyl alcohol
5.0
c) Stearic acid 8.0
d) Squalane
10.0
e) Self-emulsifying glyceryl monostearate
3.0
f) Polyoxyethylene cetyl ether (20E.O.)
1.0
g) Preservatives and antioxidants
Appropriate amount
h) Purified water
The rest
i) 1,3-butylene glycol
5.0
j) Edelweiss extract

0.0001
k) Potassium hydroxide
0.3
Manufacturing method: Heat up to a) to g) and keep at 80 ° C Heat up to h) to k), keep at 80 ° C., add to a) to g), emulsify, and cool to 40 ° C. with stirring.

(2) Emulsion composition
(weight%)
a) Beeswax
0.5
b) Petrolatum
2.0
c) Squalane
8.0
d) Sorbitan sesquioleate
0.8
e) Polyoxyethylene oleyl ether (20E.O.)
1.2
f) Preservatives and antioxidants
Appropriate amount
g) Purified water
The rest
h) 1,3-butylene glycol
7.0
i) Carboxyvinyl polymer
0.2
j) Edelweiss extract
1.0
k) Potassium hydroxide 0.1
l) Ethanol
7.0
Manufacturing method: Heat up to a) to f) and keep at 80 ° C. G) to j) are heated and dissolved, kept at 80 ° C., added to a) to f) and emulsified, and k) previously dissolved in part of g) is added and mixed uniformly. Then add l) at 50 ° C. and cool to 40 ° C.

(3) Lotion-like composition (wt%)
a) Edelweiss extract
20.0
b) α-Linolenic acid

0.1
c) Glycerin
5.0
d) Polyoxyethylene sorbitan monolaurate (20E.O.)
1.0
e) ethanol
6.0
f) Fragrance
Appropriate amount
g) Preservatives and antioxidants

Appropriate amount
h) Purified water
The remainder of the manufacturing method: a) to h) are mixed and dissolved uniformly.

(4) Packing agent
(weight%)
a) Edelweiss extract
0.01
b) Vinyl acetate resin emulsion
15.0
c) Polyvinyl alcohol 10.0
d) Cuui seed oil
3.0
e) Glycerin
5.0
f) Titanium oxide
8.0
g) Kaolin
7.0
h) ethanol
8.0
i) Fragrance
Appropriate amount
j) Preservatives and antioxidants
k) Purified water
Mix the rest of the production methods a) to k), stir and disperse well, and make uniform.

(5) Ointment
(weight%)
a) Liquid paraffin 20.0
b) White petrolatum 50.0
c) Cetanol 10.0
d) Polyoxyethylene (60) hydrogenated castor oil 18.0
e) Edelweiss extract 0.1
Production method: a) to e) are added and mixed while heating to prepare an ointment.

(6) Gel agent
(weight%)
a) Polyoxyethylene (60) hydrogenated castor oil 9.0
b) Carbopol 941 0.5
c) Ethanol 22.0
d) Triethanolamine 1.5
e) Edelweiss extract 5.0
f) Purified water 63.0
Production method: While heating, add a), b) and e) to f) and dissolve, then add d) and c) and cool with stirring to prepare a gel.

(7) Liquid agent
(weight%)
a) Polyoxyethylene (60) hydrogenated castor oil 10.0
b) Ethanol 20.0
c) Edelweiss extract 10.0
d) Purified water 60.0
Production method: a) to d) are added and dissolved to prepare a solution.

By blending the edelweiss extract with the involcrine production promoting effect of the present invention, keratinization of the stratum corneum cells is promoted, and the barrier function of the stratum corneum is improved. It can be widely applied to skin preparations.

Claims (2)

An involucrin production promoter characterized by comprising, as an active ingredient, an extract obtained from Gnaphalium uliginosum L., a plant belonging to the genus Glephalium. A skin barrier function-improving agent comprising an extract obtained from Gnaphalium uliginosum L. of an Asteraceae Lenatopodium genus (Gnaphalium) as an active ingredient.