JP2002201125A - Suppressing agent or repairing agent for skin disorder by dryness - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a skin care preparation for suppressing or repairing skin disorders caused by dryness in relation to the external preparation for cosmetic or dermatology. SOLUTION: This skin care preparation comprises thiotaurine as an active ingredient and is for suppressing or repairing reduction of an expression of filaggrin gene.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a cosmetic or dermatological external preparation, and more specifically to a skin external preparation for suppressing or repairing skin damage due to drying.

[0002]

2. Description of the Related Art Natural moisturizing factors (NM)
F) plays an important role (BlankI.HJI. Dermato
l., 18, 433 (1952), Blank IHJI. Derm
atol., 21, 259 (1953)). It has been reported that amino acids, which are the main components of NMF, are produced by the degradation of filaggrin protein derived from keratohyalin granules (Scott IR et al. Biochem Biophys. Act).
a., 719, 110-117 (1982), Horii I. et.
al., J. Dermatol., 10, 25-33 (198
3)). Filaggrin is a protein consisting of 317 amino acids. Since it was revealed that the amino acid which is the main component of NMF is derived from filaggrin, research on the relationship between filaggrin and a disease state exhibiting dry skin has been advanced. In recent years, amino acids in the stratum corneum have been reduced in dry skin such as senile xeroderma and atopic disease (Horii I. et al., Br. J. Dermatol., 121,
587-592 (1989), Tanaka M. et al., B
r. J. Dermatol., 139, 618-621 (199
8)) and decreased expression of filaggrin (Tezuka T. et al., Dermatology, 188, 21-2).
4 (1994), Seguchi T, et al., Arch. Dermatol.
Res. 288, 442-446, (1996)).

It is well known that a dry environment causes skin troubles such as rough skin.

[0004] From such a background, FR27771
85-A1 proposes a composition for the treatment of wrinkles in which a substance stimulating filaggrin synthesis is combined with other active substances to favorably regulate the water retention capacity. However, there is no description about what kind of substance stimulates filaggrin synthesis. In addition, JP-A-2
Japanese Patent Application Publication No. 000-26272 discloses that a proteolytic product derived from white lupine [or Lupinus albus ] seed powder increases the mRNA of human keratinocyte filaggrin and increases the thickness of the stratum corneum. There is a statement that there is. In addition, WO99 / 4
7117 describes that a composition containing a vitamin B3 compound acts as a humectant and increases the level of skin proteins selected from filaggrin, keratin and invocrine, and enhances the ability of water to adsorb or absorb into the stratum corneum. Have been.

On the other hand, it has been reported that peroxidation of epidermal lipids is enhanced in atopic diseases exhibiting dry skin (Kono et al., Oil Chemistry, 44, 248-255 (199).
5)), however, no report has been made on the correlation between the enhancement of skin oxidation and the decrease in water content of the stratum corneum.

[0006]

As described above, substances that increase the production of filaggrin and increase the thickness of the stratum corneum, or substances that increase the ability to absorb moisture into the stratum corneum, and which have a moisturizing action. Have been proposed as active ingredients in cosmetics.

[0007] However, it would still be desirable to provide substances or compounds that are involved in the production of filaggrin and that are effective from a broader perspective in maintaining a healthy skin condition.

[0008]

Means for Solving the Problems The present inventors have studied the factors involved in drying and the water content of the stratum corneum in human keratinocytes. As a result, the present inventors have found that the level of filaggrin protein and the expression of the gene are reduced by drying. I found it. In addition, they found a new finding that oxidation of human keratinocytes is enhanced by drying. In other words,
It was revealed that oxidative stress is involved as a mediator of skin injury due to drying. That is, it has been revealed that skin damage accompanying the decrease in filaggrin gene expression due to drying is caused not only by drying but also by oxidation of human keratinocytes. This suggests that a wide variety of substances having an antioxidant effect, such as compounds having various chemical structures and compositions that are expected to be composed of components different from these compounds, cause the filaggrin gene in human keratinocytes produced by drying. This is supported by the ability to significantly prevent the decrease in expression. In addition, the present inventors have found that thiotaurine is preferable among these substances having an antioxidant effect.

Therefore, according to the present invention, there is provided an external preparation for skin for suppressing or repairing a decrease in expression of a filaggrin gene comprising thiotaurine as an active ingredient.

According to a preferred embodiment of the present invention, the skin comprising thiotaurine as an active ingredient, and wherein the expression of the filaggrin gene is an event in human keratinocytes, and / or a decrease in the expression of the filaggrin gene is caused by dry skin. An external preparation is provided.

[0011]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The thiotaurine used in the external preparation for skin of the present invention is, for example, International Cosmetic I.
ngredient Dictionary and Handbook, 7ed. 199
7. The Cosmetic, Toiletry, and Fragrance Associat
As described in Ion, USA, as an antioxidant, it is a known substance and is a compound represented by the following formula: H ₂ NCH ₂ CH ₂ SO ₂ SH. According to the invention, the above-mentioned thiotaurine can also be used for the purposes of the invention in combination with other compounds having an antioxidant action, provided that they are not adversely affected. Such other compounds having an antioxidant action include, but are not limited to, ascorbic acid and salts or derivatives thereof, for example, ascorbic acid itself, magnesium ascorbate, sodium ascorbate, alkyl ascorbate, ascorbine Ascorbic acid derivatives such as acid phosphate, ascorbic acid sulfate, and ascorbic acid glucoside. More specifically, ascorbic acid alkyl ester, ascorbyl palmitate, ascorbyl isopalmitate, ascorbyl dipalmitate, ascorbyl diisopalmitate,
Ascorbyl stearate, ascorbyl isostearate, ascorbyl distearate, ascorbyl myristate, ascorbyl isomyristate, ascorbyl dimyristate, ascorbyl diisomyristate,
Ascorbyl 2-ethylhexanoate, ascorbyl di-2-ethylhexanoate, ascorbyl oleate, ascorbyl dioleate, and the like, as other derivatives, ascorbate polypeptide, magnesium ascorbyl phosphate, methylsilanol ascorbate, Ascorbyl tocopheryl potassium phosphate and the like can be mentioned.

Further, other compounds having an antioxidant activity include tocopherol and derivatives thereof, and the derivatives include, but are not limited to, tocopheryl esters such as tocopheryl acetate, tocopheryl linoleate, tocopheryl nicotinate, and tocopheryl succinate. And tocopherols such as tocopheres-5, tocopheres-10 and tocopheres-12. Further, as other compounds having an antioxidant effect, compounds containing sulfur-containing amino acids (including sulfur-containing amino acids themselves) and their intermediate metabolites, for example, acetylcysteine, methionine, cysteine, homocysteine, glutathione, hypotaurine , Cysteine sulfinic acid, cysteic acid, thiocysteine, taurine, diencholate, cystathionine, S-allyl cysteine, lenthionine, ethionine and the like.

In addition, thiotaurine can be included in the external preparation for skin of the present invention in addition to the above-mentioned other compounds having an antioxidant action, or individually together with a humectant. Such humectants include, for example, inositol, lactitol, maltitol, mannitol, xylitol,
Sugar alcohols such as erythritol; polyethylene glycol having a degree of polymerization of 4 to 40; glycerin, diglycerin,
1,3-butylene alcohol; propylene glycol,
Urea, lactic acid and the like can be mentioned.

In the external preparation for skin of the present invention, thiotaurine as an active ingredient can be contained in an amount of 0.0001 to 20% by weight, preferably 0.001 to 0.1% by weight, based on the total weight of the preparation. In any case, based on the results obtained in accordance with the test method or evaluation method described below for each specific use mode, and further considering the results of actual use by volunteers and the like on a trial basis, it is possible to obtain specialized knowledge in the field of dermatology. Homes will readily be able to determine the optimal dosage for the active ingredients to work.

[0015] The external preparation for skin of the present invention includes cosmetics, pharmaceuticals,
Quasi-drugs, those applied to the hull, and therefore the dosage form is also aqueous, solubilizing, emulsifying, powder, oil-liquid,
It can take a wide variety of forms, such as a gel system, an ointment system, a cream, a water-oil two-layer system, a water-oil-powder three-layer system. That is, basic cosmetics can be widely applied to various forms such as face wash, lotion, milky lotion, cream, gel, essence (serum), pack, mask and the like. If it is a makeup cosmetic, it can be widely applied to forms such as a foundation. Furthermore, if it is a medicine or a quasi-drug, it can be widely applied to various ointments and the like. The form and form of the external preparation for skin of the present invention are not limited to these forms and forms.

The base component can be selected according to the dosage form and form, and the base component or additive may be a liquid oil, a solid oil, a wax, an ester oil, a hydrocarbon oil, Silicone resin, silicone, anionic surfactant, anionic water-soluble polymer, cationic surfactant, amphoteric surfactant, nonionic surfactant,
Examples include lower alcohols, sterols, water-soluble polymers, sequestering agents, neutralizing agents, pH adjusters, antibacterial agents, fragrances, dyes, and the like.

[0017]

Now, the present invention will be described in further detail with reference to specific examples. The percentages described in the formulations are based on weight unless otherwise specified.

Test method-Determination of filaggrin gene expression level: (1) Cultured cells Normal keratinocytes derived from human foreskin and 3T derived from mouse
Three cells were used. (2) Culture medium for cultured cells The culture medium contains DMEM-Ham's F12 (3: 1), hydrocortisone, cholera enterotoxin, epidermal growth factor, insulin and 10% FBS. (3) Culture In this culture, normal cells derived from human foreskin and mouse-derived 3T3 cells (AT cells) were used as feeder layers (supporting cells).
CC CRL-1658). These cell lines are described, for example, in Rheinwald et al., Cell
6, 331-344 (1975), 3T3 cells are cultured in a suitable petri dish and treated with mitomycin C. After forming the feeder cell layer in this manner, 1 × 10 ⁵ human normal keratinocytes are seeded on the layer and cultured, for example, at 37 ° C. in a 95% air-5% carbon dioxide gas environment until the cells become confluent. Thus, human keratinocytes that can be used in this measurement can be prepared.

Gas phase exposure was performed by removing the culture supernatant of the above human keratinocytes. Each test substance was added 30 minutes before the gas phase exposure, and cultivation was performed at 37 ° C. Gas phase exposure was performed at 37 ° C. for 0-6 hours. (4) Measurement of oxidation level Using the above culture system, the lipid peroxidation level of cells exposed to the gas phase for a certain period of time was measured using malondialdehyde (MDA).
hyde) as an index. It has been reported that malondialdehyde can generally be a good indicator of lipid peroxides (Esterbauer H. et al., Free Rad. Biol. Me.
d. 11, 81-128 (1991)). Cellular lipid peroxide and N-methyl-2-phenylindole
-2-phenylindole) was reacted at 45 ° C. and quantified at 586 nm. This reaction is known and is widely used for measuring lipid peroxide. (5) Measurement of filaggrin gene expression level RNA was extracted by AGPC from cells exposed to gas phase for a certain period of time after thiotaurine was not added (control) or added to the above culture system. RT-PCR was performed on these RNAs as described below. Primers specific for filaggrin were obtained from McKinley-Grant LJ et al., Proc.
tl. Acad. Sci. USA, 86, 4848-4852
(1989) based on the filaggrin base sequence reported. In RT-PCR, mRNA was once converted into cDNA using reverse transcriptase, and using that as a template, a polymerase reaction (known reaction) was performed using the above primers to measure filaggrin mRNA. The PCR product was electrophoresed on an agarose gel, and the gel after electrophoresis was immersed in an ethidium bromide solution, and then the DNA was measured. The DNA band was quantified using NIH images (analysis program). Correction was performed using glyceraldehyde-3-phosphate dehydrogenase (G3PDH). (6) Results (6-1) FIG. 1 shows the change over time in the expression level of the filaggrin gene in human keratinocytes due to the above gas phase exposure. From the figure, it is clear that the expression of filaggrin gene in human keratinocytes is significantly reduced by drying.
Further, the decrease occurs within a relatively short time. (6-2) Changes in oxidation level due to drying in human keratinocytes. Oxidation starts to accelerate in a short time after gas phase exposure, and about 7 hours before gas phase exposure
It is clear that the oxidation is enhanced. The results are shown in FIG. (6-3) The effect of suppressing injury that reduces the expression of the filaggrin gene due to drying in human keratinocytes was evaluated based on the expression level of the filaggrin gene depending on whether or not thiotaurine was added to the culture system. The results are shown in FIG. Further, the concentration dependency is also shown.

From these results, it can be seen that the preparation according to the present invention can reduce the decrease in the water content of the stratum corneum caused by drying of human keratinocytes, the amount of the protein of filaggrin which is a raw material of amino acids essential for maintaining the water content of the stratum corneum, and the amount of the gene. It is confirmed that the decrease in expression can be significantly suppressed.

Hereinafter, formulation examples of the preparation according to the present invention will be shown. Formulation Example 1 Behenyl alcohol 1.0% Stearyl alcohol 2.0% Squalane 10.0% Pentaerythyl tetraoctanoate 5.0% Lanolin 5.0% Paraben 0.3% Polyoxyethylene behenyl alcohol 1.0% Glycerin 10. 0% Thiotaurine 0.1% Sodium acetyl hyaluronate 0.1% Purified water Remaining Formulation Example 2 Squalane 10.0% Isopropyl myristate 20.0% Decamethylcyclopentanehexane 35.0% Alkyl-modified carboxyvinyl polymer 0.05 % Sorbitan monooleate 5.0% Dipropylene glycol 3.0% Thiotaurine 1.0% Purified water Remaining formulation Example 3 Liquid paraffin 10.0% Octamethylcyclotetrasiloxane 5.0% 1,3-butyl glycol 10.0 Glycerin 3.0% Ascorbi Acid derivative 3.0% Thiotaurine 0.1% Carboxyvinyl polymer 0.15% Xanthan gum 0.25% Stearic acid 1.5% Cetyl alcohol 0.5% Polyoxyethylene (10) monooleate 1.0% Glycerin Monostearic acid ester 1.0% Ethanol 3.0% Perfume Appropriate amount Purified water Remaining Formulation Example 4 Cetostearyl alcohol 3.5% Squalane 20.0% Beeswax 3.0% Lanolin 5.0% Paraben 0.3% Polyoxy Ethylene (20) sonobitan monopalmitate 2.0% Stearic acid monoglyceride 2.0% Thiotaurine 0.1% Vitamin A 2.0% Fragrance Appropriate amount Purified water balance Prescription example 5 Talc 3.0% Titanium dioxide 5.0 % Bengala 0.5% Yellow iron oxide 1.4% Black iron oxide 1.0% Polyoxyethylene sorby monostearate Tan 0.9% Triethanolamine 1.0% Propylene glycol 5.0% Thiotaurine 1.0% Stearic acid 2.2% Isohexadecyl alcohol 7.0% Stearic acid monoglyceride 2.0% Lanolin 2.0% Paraffin 8.0% Paraben suitable amount perfume proper amount

[Brief description of the drawings]

FIG. 1 is a graph showing changes in the expression level of the filaggrin gene over time due to drying (gas phase exposure) of cultured human keratinocytes. Mean ± SE in trials n = 4-6
M is displayed. Note that ** indicates p <0.01 and * indicates p <0.05.

FIG. 2 is a graph showing the degree of enhancement of oxidation due to drying (gas phase exposure) of cultured human keratinocytes. Malondialdehyde (MDA) was used as an index. The average per 10 dishes is shown. The unit is μmol / g protein.

FIG. 3 is a graph showing the effect of suppressing the decrease in thiotaurine on the decrease in the expression level of filaggrin due to drying (gas phase exposure) of cultured human keratinocytes. Displayed according to the mean ± SEM of trial n = 3. ** indicates p <
Means 0.01.

Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat II (Reference) A61P 43/00 A61P 43/00 (72) Inventor Tetsuji Hirao 2-12-1 Fukuura, Kanazawa-ku, Yokohama-shi, Kanagawa Corporation Shiseido Research Center F-term (reference) 4C083 AA082 AB232 AB242 AB442 AC012 AC022 AC072 AC092 AC102 AC112 AC122 AC182 AC242 AC352 AC402 AC422 AC442 AC482 AC542 AC771 AC772 AD092 AD172 AD272 AD332 AD352 AD512 AD622 AD642 4C206 AA01 AA02 JA78 MA78

Claims (3)

[Claims] 1. An external preparation for skin for suppressing or repairing a decrease in expression of a filaggrin gene comprising thiotaurine as an active ingredient. 2. The external preparation for skin according to claim 1, wherein the expression of the filaggrin gene is an event in human keratinocytes. 3. The external preparation for skin according to claim 1, wherein the decrease in the expression of the filaggrin gene is caused by drying of the skin.