JP4426919B2 - β4 integrin production promoter and whitening cosmetic. - Google Patents

Description

  The present invention relates to a whitening cosmetic comprising a component that promotes the production of β4 integrin, which is a component of hemidesmosome.

The mechanism of stain formation has not yet been fully clarified, but it is thought that increased melanin production in pigment cells and delayed melanin excretion due to decreased epidermal turnover are thought to cause this condition. β4 integrin is one of the constituent molecules of hemidesmosome responsible for adhesion between epidermal basal cells and basement membrane, and plays an important role in maintaining the structure of normal skin (see Non-Patent Document 1). It also has a function of transmitting a signal into cells and is involved in the regulation of epidermal turnover (see Non-Patent Documents 2 and 3). However, sufficient knowledge has not been obtained as to whether and how β4 integrin is involved in the decrease in epidermal turnover at the spot site. Conventionally, dichloroacetic acid has been known as a component having cell activation activity against epidermal keratinocytes (see Non-Patent Document 4), but it is known what effect is produced when it is formulated in cosmetics. It wasn't.
Vidal F et al: Nature Genet, 1995, Vol. 10, p229-234 Dawling J et al: J Cell Biol, 1996, 134, p559-572. Mainiero F et al: EMBO J, 1997, 16, p2365-2375 Kitamura N et al: Skin Pharmacol Appl Skin Physiol, 1999, Vol. 12, p317-325

  This invention pays attention to the phenomenon in a spot affected part, and is providing the whitening cosmetics based on the new mechanism which concerns on the turnover of an epidermis.

  As a result of various analyzes of the skin tissue at the spot site, the present inventor has found that the expression of β4 integrin is reduced at the spot site. And it discovered that a specific chlorine substituted alcohol, aldehyde, or carboxylic acid had a novel function as a β4 integrin production promoter that promotes the production of β4 integrin. Furthermore, when this β4 integrin production promoter was blended in cosmetics, it was found that it had a remarkable whitening effect and the present invention was completed.

That is, the present invention resides in a β4 integrin production promoter consisting of one or more selected from the group consisting of a chlorine-substituted alcohol, aldehyde or carboxylic acid, salt or ester thereof represented by R (Cl) -X (wherein R (Cl) represents a methyl group or an ethyl group in which at least one hydrogen atom is substituted with a chlorine atom, X is —CH ₂ OH, —CHO or —COOY, Y is a hydrogen atom, an alkali metal, An alkaline earth metal, ammonium, organic ammonium, or an alkyl group having 3 or less carbon atoms).

  The second invention of the present application is a whitening cosmetic comprising the β4 integrin production promoter.

  The β4 integrin production promoter consisting of a specific chlorine-substituted alcohol, aldehyde or carboxylic acid according to the present invention promotes the production of β4 integrin in the epidermal basal cell and normalizes the adhesion between the basal cell and the basement membrane. And the whitening cosmetic containing a β4 integrin production promoter improves the epidermis turnover, which is reduced in the affected area, and exhibits a whitening effect.

The β4 integrin production promoter in the present invention is at least one selected from the group consisting of a chlorine-substituted alcohol represented by R (Cl) -X, an aldehyde or a carboxylic acid, a salt or an ester thereof (wherein R ( Cl) represents a methyl group or an ethyl group in which at least one hydrogen atom is substituted with a chlorine atom, X represents —CH ₂ OH, —CHO or —COOY, and Y represents a hydrogen atom, an alkali metal, an alkaline earth, or the like. A similar metal, ammonium, organic ammonium, or an alkyl group having 3 or less carbon atoms). As the organic ammonium, methylammonium, dimethylammonium, trimethylammonium, ethylammonium, monoethanolammonium, diethanolammonium, triethanolammonium, monopropanolammonium, diisopropylammonium and the like can be used. These compounds can be synthesized by a conventionally known method. It can also be obtained as a commercial product.

  The β4 integrin production promoter of the present invention has an action of promoting the production of β4 integrin in epidermal cells. By adding it to cosmetics, the skin turnover lowered at the spot site is improved and a whitening effect is obtained. The whitening cosmetic composition of the present invention can be achieved.

  The blending amount of the β4 integrin production promoter in the whitening cosmetic composition of the present invention is preferably 0.001 to 5.0 mass%, more preferably 0.01 to 1.0 mass%. If the blending amount is less than 0.01% by mass, a sufficient effect cannot be obtained, and even if it exceeds 5.0% by mass, an effect commensurate with it cannot be obtained.

  The whitening cosmetic composition of the present invention, in addition to the above-described β4 integrin production promoter, which is an essential constituent, is arbutin, ellagic acid, oil-soluble licorice extract, retinoid, phenolic dimer compound, 4-n-butylresorcinol, Further whitening by appropriately blending at least one selected from whitening ingredients such as ellagic acid, tranexamic acid, chamomile extract, linoleic acid, α-hydroxy acid, hydroquinone, cysteine, glutathione, vitamin E, vitamin C derivatives, etc. Expected to be effective. Among these, arbutin, ellagic acid, 4-n-butylresorcinol and vitamin C derivatives are particularly preferably used, and the concentration in the whitening cosmetic may be 0.01% by mass or more.

  Furthermore, the whitening cosmetics of the present application include other components such as an anionic surfactant, an amphoteric surfactant, a nonionic surfactant, a sticky agent, an oil agent, and a powder as long as the object of the present invention is not impaired. (Pigments, resins, pigments), preservatives, fragrances, moisturizers, physiologically active ingredients, salts, solvents, antioxidants, chelating agents, pearling agents, neutralizing agents, pH adjusters, insect repellents, enzymes, etc. Components can be appropriately blended. Although the specific example of a mixing | blending component is shown below, it is not restricted to these.

Anionic surfactants include α-acyl sulfonate, alkyl sulfonate, alkyl allyl sulfonate, alkyl naphthalene sulfonate, alkyl sulfate, alkyl ether sulfate, alkyl amide sulfate, polyoxyethylene Alkyl ether sulfates, polyoxyethylene alkylamide ether sulfates, alkyl phosphates, alkylamide phosphates, alkyloylalkyl taurine salts, N-acyl amino acid salts, sulfosuccinates, perfluoroalkyl phosphates, etc. It is done.

  Examples of amphoteric surfactants include glycine type, aminopropionic acid type, carboxybetaine type, sulfobetaine type, sulfonic acid type, sulfuric acid type, and phosphoric acid type, and 2-alkyl-N-carboxymethyl is preferable. -N-hydroxyethyl imidazolinium betaine, coconut oil fatty acid amidopropyl betaine, etc. can be illustrated.

  Nonionic surfactants include fatty acid alkanolamide, polyoxyethylene hydrogenated castor oil, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene alkyl ether, polyoxyethylene alkyl ester, sucrose fatty acid ester, polyglycerin fatty acid ester, alkyl An amine oxide etc. are mentioned.

  Examples of the adhesive include acrylic amide and derivatives thereof, carboxyvinyl polymer, alkyl-modified carboxyvinyl polymer, cellulose, keratin and collagen or derivatives thereof, calcium alginate, pullulan, agar, gelatin, tamarind seed polysaccharide, xanthan gum, carrageenan , High methoxyl pectin, low methoxyl pectin, guar gum, gum arabic, crystalline cellulose, arabinogalactan, karaya gum, tragacanth gum, alginic acid, albumin, casein, curdlan, gellan gum, dextran and the like.

  Examples of the oil agent include volatile and non-volatile oil agents, solvents, and resins that are usually used in cosmetics, and may be liquid, paste, or solid at room temperature. Examples of oils include higher alcohols such as cetyl alcohol, isostearyl alcohol, lauryl alcohol, hexadecyl alcohol, octyldodecanol, fatty acids such as isostearic acid, undecylenic acid, oleic acid, myristyl myristate, hexyl laurate, myristine Octyldodecyl acid, decyl oleate, isopropyl myristate, hexyl decyl dimethyloctanoate, glyceryl monostearate, glyceryl trioctanoate, diethyl phthalate, ethylene glycol monostearate, octyl oxystearate, cholesteryl stearate, Cholesterol esters such as cholesteryl oleate and branched fatty acid cholesteryl, hydrocarbons such as liquid paraffin, petroleum jelly and squalane, lanoli Waxes such as reduced lanolin, carnauba wax, mink oil, cacao butter, palm oil, palm kernel oil, camellia oil, sesame oil, castor oil, olive oil, dimethylpolysiloxane, cyclic dimethylpolysiloxane, methylphenylpolysiloxane, etc. Silicone oil etc. are mentioned.

  Examples of powders include red No. 201, yellow No. 4, blue No. 1, black No. 401 and other dyes, yellow No. 4 Al lake, yellow No. 203 Ba lake and other lake dyes, nylon powder, silk powder and silicone powder. , Cellulose powder, silicone elastomer spherical powder, resin such as polyethylene powder, yellow iron oxide, red iron oxide, chromium oxide, carbon black, colored pigments such as ultramarine and bitumen, white pigments such as zinc oxide and titanium oxide, talc, Body pigments such as mica, sericite, kaolin, pearl pigments such as titanium mica, metal salts such as barium sulfate, calcium carbonate, magnesium carbonate, magnesium silicate, inorganic powders such as silica and alumina, bentonite, smectite, nitriding Boron etc. are mentioned. There are no particular restrictions on the shape of these powders (spherical, rod-like, needle-like, plate-like, indeterminate, flake-like, spindle-like, etc.).

Examples of the physiologically active ingredient include substances that impart some physiological activity to the skin when applied to the skin. For example, anti-aging agents, UV protection agents, squeeze agents, antioxidants, moisturizers, blood circulation promoters, antibacterial agents, bactericides, desiccants, cooling agents, warming agents, vitamins, amino acids, wound healing promoters , Stimulation mitigating agents, analgesics, cell activators, enzyme components and the like.

  The whitening cosmetic composition of the present invention can be produced according to a conventional method. The cosmetics of the present invention are not limited to general skin cosmetics, but include quasi drugs, designated quasi drugs, topical drugs, etc., and their dosage forms are also particularly limited. However, it can be arbitrarily selected according to the purpose. That is, dosage forms such as creams, ointments, emulsions, solutions, gels, and packs, lotions (skin lotions), powders, sticks, and the like can be used.

EXAMPLES Hereinafter, although an Example is given and this invention is further demonstrated, this invention is not limited to these Examples. In addition, unless otherwise indicated, the unit of the compounding quantity in an Example is the mass%.
[Expression of β4 integrin in human epidermis]
Normal human skin and stain skin tissue were frozen and embedded, and 5 μm slices were prepared. After the section was fixed with acetone, antibody staining was performed using an anti-human β4 integrin antibody (Chemicon) as a primary antibody. VECTASTAIN ABC kit (VECTOR) and DAB substrate kit (Funakoshi) were used for detection.

The expression of β4 integrin in human epidermis is shown in FIG. 1 and FIG. In normal skin (FIG. 1), the expression of β4 integrin was confirmed in epidermal basal cells continuously along the basement membrane, which is the boundary between the epidermis and dermis. On the other hand, it was confirmed that the expression level of β4 integrin in epidermal basal cells was reduced in the blemishes (FIG. 2) having excessive melanin.
[Evaluation of β4 integrin expression promoting effect]
1. Human neonatal foreskin-derived epidermal keratinocytes (purchased from Kurabo Corporation) were used.
2. Epidermal keratinocytes were prepared in a medium so as to be 5.5 × 10 ⁴ cells / ml, and 5 ml each was seeded on a 6 cmφ culture plate (Falcon). The medium used was MCDB153 medium supplemented with BPE (bovine pituitary extract), insulin, ethanolamine, phosphoethanolamine, and hydrocortisone as growth factors. The cells were pre-cultured to a confluent state at 37 ° C. in a 5% (V / V) CO ₂ atmosphere.
3. The medium supernatant was removed by aspiration, the medium was replaced with a medium to which the test sample was added at a specified concentration, and the cells were cultured at 37 ° C. for 72 hours in a 5% (V / V) CO ₂ atmosphere. The medium used was MCDB153 medium supplemented with insulin, ethanolamine, and phosphoethanolamine as growth factors. 4). After incubation, total RNA was extracted with an RNA purification kit (Qiagen). After performing reverse transcription reaction using Super Script II reagent (Invitrogen), PCR reaction was performed using real-time PCR analysis system (ABI PRISM 7000, Applied Biosystems), and β4 integrin mRNA expression level was examined. . As a PCR probe for detecting β4 integrin, a primer set sold by Applied Biosystems was used, and the PCR reaction conditions followed the protocol. At the same time, the expression level of G3PDH mRNA, which is a housekeeping gene, was quantified, and the expression level of β4 integrin was evaluated by β4 integrin mRNA / G3PDH mRNA.

The production promoting action of the β4 integrin production promoter of the present invention was evaluated by the above evaluation system. Dichloroacetic acid, trichloroacetic acid and chloroacetic acid used for evaluation were purchased from Wako Pure Chemical Industries, and dichloroethanol was purchased from Sigma-Aldrich.
Example 1 (dichloroacetic acid)
FIG. 3 shows the expression level of β4 integrin when dichloroacetic acid is added as a β4 integrin production promoter at different concentrations. Addition of dichloroacetic acid promoted the expression of β4 integrin mRNA in a concentration-dependent manner. In addition, G3PDH mRNA was added by adding dichloroacetic acid
There was no change in the expression level.
Example 2 (dichloroacetic acid, trichloroacetic acid, chloroacetic acid, dichloroethanol)
FIG. 4 shows the expression level of β4 integrin when dichloroacetic acid, trichloroacetic acid, chloroacetic acid and dichloroethanol are added at 100 μmol / l. Trichloroacetic acid promoted the expression of β4 integrin mRNA at the same level as dichloroacetic acid. Chloroacetic acid and dichloroethanol had higher β4 integrin production promoting activity, which was about twice as high as dichloroacetic acid and trichloroacetic acid.

The effect of the whitening cosmetic containing the β4 integrin production promoter of the present invention was evaluated by the following whitening practical test.
[Whitening test method]
An appropriate amount of the evaluation sample was applied twice a day for the affected area of 10 test subjects having spots on the face. The application period was at least 7 weeks. Based on the questionnaire results, the number of subjects who answered that the state of the spots at each application site had improved compared to before the start of the test was used as an evaluation index in the whitening practical test.
Example 3, Comparative Example 1: Lotion According to the raw material composition and production method shown in Table 1 below, the lotion of Example 3 was prepared and a whitening practical test was conducted. In addition, the lotion of Comparative Example 1 was obtained without adding C component.
(Table 1)
――――――――――――――――――――――――
Ingredients Components Blending amount
A component
(1) Ethanol 8.0
(2) Polyoxyethylene monolaurate 0.5
Sorbitan (20E.O.)
(3) Fragrance 0.01
(4) Methylparaben 0.1
B component
(5) Glycerin 3.0
(6) 1,3-butylene glycol 1.0
(7) Carrageenan 0.2
(8) Citric acid 0.01
(9) Sodium citrate 0.09
(10) Diisopropylamine 0.25
(11) Purified water remaining C component (β4 integrin production promoter)
(12) Dichloroacetic acid Listed in Table 2 ――――――――――――――――――――――――
(Production method)
After mixing and dissolving the A component and B component shown in Table 1, the B component was added to the A component and mixed and stirred, and then the C component was added and mixed and stirred.
(Whitening practical test results)
The results of the whitening practical test are shown in Table 2. As shown in Table 2, the whitening cosmetic (lotion) of Example 3 of the present invention containing a component that promotes the production of β4 integrin clearly showed good results.
(Table 2)
――――――――――――――――――――――――
Evaluation sample Concentration Whitening practical test ――――――――――――――――――――――――
Example 3 (dichloroacetic acid) 0.25 7
Comparative Example 1 (-)-1
――――――――――――――――――――――――
In addition, the whitening cosmetic of Example 3 did not develop redness, inflammation, or other symptoms considered to be side effects on the skin, indicating that the whitening cosmetic according to the present invention is also excellent in safety.
Example 4, Comparative Example 2: Cream A cream was prepared according to the raw material composition and production method shown in Table 3 below. In addition, the cream which does not mix | blend D component was made into the cream of the comparative example 2.
(Table 3)
――――――――――――――――――――――――
Ingredients Component Blending amount ――――――――――――――――――――――――
A component
(1) Glyceryl monostearate 2.0
(2) Stearic acid 1.0
(3) Behenyl alcohol 0.7
(4) Polyoxyethylene 1.0
Cetyl ether (2E.O.)
(5) Vaseline 1.5
(6) Olive Squalane 6.0
(7) Methyl phenyl silicon 4.0
(8) Cyclopentasiloxane 3.0
(9) Butylparaben 0.05
(10) Diisopropylamine 0.25
B component
(11) N-stearoyl 0.9
-Sodium L-glutamate
(12) Xanthan gum 0.2
(13) Methylparaben 0.1
(14) Purified water remaining C component
(15) Ethanol 5.0
(16) Phenoxyethanol 0.3
D component (β4 integrin production promoter)
(17) Dichloroacetic acid 0.25
――――――――――――――――――――――――
(Production method)
Each component of (A) and (B) described in Table 3 is heated to 70 ° C. and completely dissolved, and then the oil phase part is mixed with the aqueous phase part and emulsified with an emulsifier. Cool the emulsion and at 50 ° C (C),
Add (D) and cool to a final temperature of 30 ° C.
(Whitening practical test results)
As a result of the whitening practical test, the whitening cosmetic (cream) of Example 4 of the present invention containing a component that promotes the production of β4 integrin is clearly superior in whitening effect and safer than Comparative Example 2. There was no problem.

  As described above, the β4 integrin production promoter of the present invention consisting of a specific compound sufficiently promotes the production of β4 integrin in epidermal basal cells, and thereby sufficiently transmits signals related to cell adhesion, survival and proliferation of the epidermal basal layer. To improve the turnover that has been lowered in the affected area of the spot, and exhibits a whitening effect. And by blending this in cosmetics, whitening cosmetics aimed at preventing, preventing and improving spots can be provided.

FIG. 3 is a micrograph of immunohistochemical staining instead of a drawing showing the expression of β4 integrin in basal cells of normal human skin. The dark part is where β4 integrin is expressed. It is the microscope picture of the immunohistochemical staining for the figure substitution which shows the expression of (beta) 4 integrin in the basal cell of human skin skin. It is a figure which shows the production promotion effect | action of (beta) 4 integrin when a human epidermal keratinocyte is processed by changing the addition density | concentration of dichloroacetic acid (DCA). It is a figure which shows the production promotion effect | action of (beta) 4 integrin when a human epidermis keratinocyte is processed with dichloroacetic acid, trichloroacetic acid, chloroacetic acid, and dichloroethanol.

Claims (2)

Β4 integrin production promoter consisting of one or more selected from the group consisting of dichloroethanol represented by CHCl ₂ —CH ₂ OH, chloroacetic acid represented by CH ₂ Cl—COOY, and salts thereof (wherein Y is a hydrogen atom, alkali shown metals, alkaline earth metals, ammonium or organic ammonium beam.).   A whitening cosmetic comprising the β4 integrin production promoter according to claim 1.