Skin Sensitivity Enhancers

Abstract

To provide an agent for activating skin sensation, an agent for promoting the activity of Merkel cells, an agent for promoting piezo2 expression in Merkel cells, an agent for promoting IGF-1 expression in Merkel cells, and an agent for promoting oxytocin expression in Merkel cell.SOLUTION: The present invention provides an agent for activating skin sensation, an agent for promoting the activity of Merkel cells, an agent for promoting piezo2 expression in Merkel cells, an agent for promoting IGF-1 expression in Merkel cells, and an agent for promoting oxytocin expression in Merkel cell, each containing a peptide that contains arginine as a constituent amino acid.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a skin sensory activator, a Merkel cell activity promoter, a piezo2 expression promoter in Merkel cells, an IGF-1 expression promoter in Merkel cells, and an oxytocin expression promoter in Merkel cells. [Background technology]

[0002] Merkel cells and Meissner's corpuscles, which are located relatively shallow in the skin, are known as sensory receptors in the skin. Each of these types of sensory receptors responds to physical stimuli, such as vibrations at the contact point, with different vibration frequencies and displacements (amplitudes). It is known that the minimum amplitude of vibrations to which Merkel cells and Meissner's corpuscles can respond is relatively large. Tactile sensation presentation devices capable of providing physical stimuli to which the sensory receptors in the skin can react have been provided (Patent Documents 1, 2, etc.). These tactile sensation presentation devices can reproduce various tactile sensations by stimulating the sensory receptors present in the skin, and are expected to be applied to fields such as virtual reality. In order to ensure a variety of tactile sensations presented to the skin, it is desirable for tactile sensation presentation devices to provide physical stimuli (vibrations) to which various sensory receptors can respectively react, and devices have been provided that provide vibrations to which sensory receptors located relatively shallow in the skin, such as Merkel cells and Meissner's corpuscles, can react. Thus, Merkel cells have attracted attention as sensory receptors in the skin, but no method for activating them other than by physical stimulation has yet been reported. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] JP 2018-010582 A [Patent Document 2] JP 2019-125310 A [Non-patent literature]

[0004] [Non-Patent Document 1] "Starting research into Merkel cells related to skin touch", [online], FANCL Corporation, April 21, 2021, press release, Internet<URL:https: / / www.fancl.jp / news / pdf / 20210421_merukerusaibou.pdf> Summary of the Invention [Problem to be solved by the invention]

[0005] Merkel cells are sensory cells that receive mechanical stimuli and were proposed by Dr. Merkel in 1875. They exist in fish to mammals and have been reported to respond to weak pressure and secrete neurotransmitters. Merkel cells are located in the basal layer of the skin, in contact with the sensory nerve endings, and are related to touch, and are found in large numbers in areas that perceive sensation, such as the fingertips. It is known that skin sensation weakens with age, but promoting the activation of Merkel cells is thought to be an effective method of maintaining and activating skin sensation. And, an objective of the present invention is to provide a skin sensory activator, a Merkel cell activity promoter, an agent for promoting piezo2 expression in Merkel cells, an agent for promoting IGF-1 expression in Merkel cells, and an agent for promoting oxytocin expression in Merkel cells. [Means for solving the problem]

[0006] The main configuration of the present invention is as follows. 1. A skin sensory activator containing a peptide that contains arginine as a constituent amino acid. 2. A Merkel cell activity promoter containing a peptide containing arginine as a constituent amino acid. 3. A piezo2 expression promoter in Merkel cells, comprising a peptide containing arginine as a constituent amino acid. 4. An agent for promoting IGF-1 expression in Merkel cells, comprising a peptide containing arginine as a constituent amino acid. 5. An agent for promoting oxytocin expression in Merkel cells, comprising a peptide containing arginine as a constituent amino acid. 6. Peptides containing arginine as a constituent amino acid include decapeptide-4, hydrolyzed yeast, and palmitoyl Tetra The enhancer according to any one of 2. to 5., which is one or more selected from the group consisting of peptide-7. Effect of the Invention

[0007] The skin sensation activator, Merkel cell activity promoter, piezo2 expression promoter in Merkel cells, IGF-1 expression promoter in Merkel cells, and oxytocin expression promoter in Merkel cells of the present invention can activate skin sensation and promote the expression of piezo2, IGF-1, and oxytocin in Merkel cells. The physiological function of Merkel cells can be improved by the skin sensory activator, Merkel cell activity promoter, Merkel cell piezo2 expression promoter, Merkel cell IGF-1 expression promoter, and Merkel cell oxytocin expression promoter of the present invention. [Brief description of the drawings]

[0008] [Figure 1] Graph showing Merkel cell activation by decapeptide-4. [Diagram 2] FIG. 1 shows the results of a test of piezo2 expression in Merkel cells by adding peptides. [Diagram 3] FIG. 1 shows the results of a test on IGF-1 expression in Merkel cells by addition of peptides. [Figure 4] FIG. 1 shows the results of a test of oxytocin expression in Merkel cells by addition of peptides. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] The present invention relates to a skin sensory activator, a Merkel cell activity promoter, a piezo2 expression promoter in Merkel cells, an IGF-1 expression promoter in Merkel cells, and an oxytocin expression promoter in Merkel cells (hereinafter, the above-mentioned agents are collectively referred to as the agents of the present invention).

[0010] Piezo2 is a mechanically sensitive ion channel that is activated by mechanical stimuli and is involved in many physiological roles such as touch, pain, and respiration. Merkel cells functionally express piezo2 channels and convert mechanical stimuli into electrical signals through their opening. Therefore, in order to activate skin sensation, it is necessary to promote the expression of piezo2 in Merkel cells.

[0011] IGF-1 is called "insulin-like growth factor" and is a type of growth hormone with a chemical structure similar to insulin. The promotion of IGF-1 expression in Merkel cells occurs through activation of Merkel cells.

[0012] Oxytocin is a type of neuropeptide hormone secreted by touching each other, and activates neighboring and self-cells and transmits the information to the brain via the afferent vagus nerve. The present inventors discovered that oxytocin is secreted from Merkel cells (Non-Patent Document 1). The promotion of oxytocin expression in Merkel cells occurs due to the activation of Merkel cells.

[0013] In the present invention, a peptide containing arginine as a constituent amino acid is used. The peptide containing arginine as a constituent amino acid used in the present invention is not particularly limited as long as it contains arginine as a constituent amino acid, and examples thereof include decapeptide-4, hydrolyzed yeast, palmitoyl tetrapeptide-7, palmitoyl tetrapeptide-1, etc. In the present invention, the peptide containing arginine as a constituent amino acid can be used alone or in combination of two or more kinds.

[0014] Decapeptide-4 (cosmetic labeling name) is a synthetic decapeptide consisting of arginine, aspartic acid, cysteine, glutamic acid, leucine, methionine, and tyrosine. Commercially available products include CG-IDP2 BG (1,3 butylene glycol) (300 ppm) manufactured by Caregen.

[0015] Hydrolyzed yeast (labeled on cosmetics) is a synthetic decapeptide consisting of arginine, alanine, lysine, aspartic acid, glutamic acid, leucine, and histidine. Commercially available products include, for example, Yeast Peptide KF manufactured by Katakura Co-op Agri Co., Ltd.

[0016] Palmitoyl tetrapeptide-7 (cosmetic name) is a reaction product of palmitic acid and a synthetic tetrapeptide consisting of arginine, glutamine, glycine and proline residues. Commercially available products include MATRIXYL3000 manufactured by Sederma SAS.

[0017] In the agent of the present invention, the concentration of the peptide that contains arginine as a constituent amino acid is not particularly limited as long as it can achieve the effect of the present invention.For example, the peptide that contains arginine as a constituent amino acid is preferably 0.000001% by weight or more and 1.0% by weight or less, more preferably 0.00001% by weight or more and 0.1% by weight or less, and even more preferably 0.0001% by weight or more and 0.01% by weight or less, based on the whole agent of the present invention.

[0018] The agent of the present invention can be manufactured according to a commonly used formulation method, and can be made into medicine, quasi-drug, or cosmetic.For example, the agent of the present invention can be made into various dosage forms such as aqueous solution, oil, lotion, emulsion, gel, cream, ointment, plaster, cataplasm, aerosol, suppository, injection, powder, granule, tablet, pill, syrup, and troche.

[0019] The agent of the present invention can contain fats and oils such as vegetable oils, higher fatty acids, higher alcohols, silicones, anionic surfactants, cationic surfactants, amphoteric surfactants, nonionic surfactants, preservatives, sugars, sequestering agents, polymers such as water-soluble polymers, thickeners, powder components, UV absorbers, UV blocking agents, moisturizing agents such as hyaluronic acid, fragrances, pH adjusters, desiccants, etc. It can also contain other medicinal and physiologically active ingredients such as vitamins, skin activators, blood circulation promoters, resident bacteria control agents, active oxygen scavengers, anti-inflammatory agents, anticancer agents, skin whitening agents, and bactericides. It can also contain peptides that do not contain arginine as a constituent amino acid.

[0020] Examples of fats and oils include liquid fats and oils such as camellia oil, evening primrose oil, macadamia nut oil, olive oil, rapeseed oil, corn oil, sesame oil, jojoba oil, germ oil, wheat germ oil, triglycerin, and glycerin trioctanoate; solid fats and oils such as cacao butter, coconut oil, hydrogenated coconut oil, palm oil, palm kernel oil, Japan wax, Japan wax kernel oil, hydrogenated oil, and hydrogenated castor oil; waxes such as beeswax, candelilla wax, cotton wax, bran wax, lanolin, lanolin acetate, liquid lanolin, and sugar cane wax; liquid paraffin, squalene, squalane, and microcrystalline wax. Examples of higher fatty acids include lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA). Examples of higher alcohols include straight-chain alcohols such as lauryl alcohol, stearyl alcohol, cetyl alcohol, and cetostearyl alcohol, and branched-chain alcohols such as monostearyl glycerin ether, lanolin alcohol, cholesterol, phytosterol, and octyldodecanol. Examples of silicones include linear polysiloxanes such as dimethylpolysiloxane and methylphenylpolysiloxane, and cyclic polysiloxanes such as decamethylpolysiloxane.

[0021] Examples of anionic surfactants include fatty acid salts such as sodium laurate, higher alkyl sulfate salts such as sodium lauryl sulfate, alkyl ether sulfate salts such as POE lauryl sulfate triethanolamine, N-acylsarcosinic acid, sulfosuccinate salts, and N-acylamino acid salts. Examples of cationic surfactants include alkyltrimethylammonium salts such as stearyltrimethylammonium chloride, benzalkonium chloride, and benzethonium chloride. can be. Examples of amphoteric surfactants include betaine-based surfactants such as alkyl betaines and amido betaines. Examples of nonionic surfactants include sorbitan fatty acid esters such as sorbitan monooleate, and hydrogenated castor oil derivatives.

[0022] Examples of preservatives include methylparaben and ethylparaben. Examples of the sequestering agent include edetic acid, edetic acid sodium salt, and the like. Examples of polymers include gum arabic, tragacanth gum, galactan, guar gum, carrageenan, pectin, agar, quince seed, dextran, pullulan, carboxymethyl starch, collagen, casein, gelatin, methylcellulose, methylhydroxypropylcellulose, hydroxyethylcellulose, sodium carboxymethylcellulose (CMC), sodium alginate, vinyl polymers such as carboxyvinyl polymers (CARBOPOL, etc.), and bentonite. Examples of thickeners include carrageenan, tragacanth gum, quince seed, casein, dextrin, gelatin, CMC, hydroxyethyl cellulose, hydroxypropyl cellulose, carboxyvinyl polymer, guar gum, and xanthan gum. Examples of the powder component include talc, kaolin, mica, silica, zeolite, polyethylene powder, polystyrene powder, cellulose powder, inorganic white pigments, inorganic red pigments, pearl pigments such as titanium oxide coated mica, titanium oxide coated talc, and colored titanium oxide coated mica, and organic pigments such as Red No. 201 and Red No. 202.

[0023] Examples of the ultraviolet absorber include para-aminobenzoic acid, phenyl salicylate, isopropyl para-methoxycinnamate, octyl para-methoxycinnamate, and 2,4-dihydroxybenzophenone. Examples of ultraviolet blocking agents include titanium oxide, talc, carmine, bentonite, kaolin, and zinc oxide. Moisturizing agents include polyethylene glycol, propylene glycol, dipropylene glycol, 1,3-butylene glycol, glycerin, diglycerin, polyglycerin, xylitol, maltitol, maltose, sorbitol, glucose, fructose, sodium chondroitin sulfate, sodium hyaluronate, sodium lactate, pyrrolidone carboxylic acid, and cyclodextrin.

[0024] Examples of medicinal ingredients include vitamins such as vitamin A oil, vitamin A such as retinol, vitamin B2 such as riboflavin, vitamin B6 such as pyridoxine hydrochloride, vitamin C such as L-ascorbic acid, L-ascorbic acid phosphate, L-ascorbic acid monopalmitate, L-ascorbic acid dipalmitate, and L-ascorbic acid-2-glucoside, pantothenic acids such as calcium pantothenate, vitamin D such as vitamin D2 and cholecalciferol, and vitamin E such as α-tocopherol, tocopherol acetate, and DL-α-tocopherol nicotinate.

[0025] Other medicinal ingredients include whitening agents such as placenta extract, glutathione, and saxifrage extract, skin activators such as royal jelly and beech wood extract, blood circulation promoters such as capsaicin, zingerone, cantharides tincture, ichthammol, caffeine, and γ-oryzanol tannate, anti-inflammatory agents such as glycyrrhizinic acid derivatives, glycyrrhetinic acid derivatives, and azulene, amino acids such as arginine, serine, leucine, and tryptophan, resident flora control agents such as maltose-sucrose condensates, and lysozyme chloride.

[0026] Further examples of extracts include chamomile extract, parsley extract, beech extract, wine yeast extract, grapefruit extract, honeysuckle extract, rice extract, grape extract, hop extract, rice bran extract, loquat extract, Phellodendron bark extract, Coix seed extract, Swertia jasmine extract, melilot extract, birch extract, licorice extract, peony extract, soapwort extract, loofah extract, chili pepper extract, lemon extract, gentiana extract, perilla extract, aloe extract, rosemary extract, sage extract, thyme extract, tea extract, seaweed extract, cucumber extract, clove extract, carrot extract, horse chestnut extract, witch hazel extract, and mulberry extract. EXAMPLES

[0027] (Cell Test) A human Merkel cell carcinoma-derived cell line (MCC14 / 2: Merkel cell carcinoma) (Cell Bank Australia) was used as an alternative to primary cultured cells in the study. This cell line has been shown to be useful for in vitro gene expression analysis, and analysis of the cytoskeleton and secretory mechanism of Merkel cells (Non-Patent Document 1). MCC14 / 2 was cultured in RPMI1640 Media (Thermo Fisher) supplemented with 10% FBS and 1x Penicillin-Streptomycin. After the cells had grown to about 80% confluence, they were detached with 0.017% Trypsin / 0.025% EDTA (0.05% Trypsin / 0.02% EDTA (Sigma-Aldrich) diluted 2-fold with PBS), resuspended, and the cells were counted by standard methods.

[0028] (Intracellular calcium ion concentration measurement) MCC14 / 2 cells were seeded on a 96-well plate, and 0.000025% concentration of decapeptide-4 was added to the cells and cultured for 48 hours. After washing the cells several times with PBS(-), a calcium ion probe was introduced into the cells using Calcium kit-Fluo4 (DOJINDO). The cells were observed using an in-cell analyzer. As a control, cells were observed in the same manner except that decapeptide-4 was not added.

[0029] (result) The results are shown in Figure 1. It was observed that decapeptide-4, a peptide containing arginine as a constituent amino acid, increased the calcium ion concentration in Merkel cells, confirming that decapeptide-4 activates Merkel cells.

[0030] (Peptide addition test) MCC14 / 2 at 2.0, 1.2, 1.0×10 5The cells were seeded on a 6-well plate at 1000x1000 cells / well. After 48 hours of seeding, decapeptide-4, hydrolyzed yeast, palmitoyl tetrapeptide-7, and palmitoyl tetrapeptide-5 were added to the cells at 0.000025% or 0.0001% by weight, respectively, and the cells were cultured for 6, 24, and 48 hours. After washing the cells with PBS(-), Tissue Protein Extraction Reagent (Thermo Fisher) was added to recover the proteins. As a control, the proteins were recovered in the same manner except that no peptide was added. Palmitoyl tetrapeptide-5 is a reaction product of a tripeptide consisting of lysine and valine with palmitic acid, and is a peptide that does not contain arginine as a constituent amino acid.

[0031] (Western blot analysis) The expression levels of piezo2, IGF-1, and oxytocin in Merkel cells produced by the addition of each peptide were measured by Western blot analysis. The method is as follows. The concentration of the collected proteins was adjusted to 0.5 M Tris-HCl (pH 6.8), 2% SDS. The proteins were separated by SDS-PAGE (5-20% gel) (DRC Co., Ltd.) and transferred to a 0.2 μm PVDF Trans-Blot Turbo Transfer Pack. The proteins were blocked at room temperature for 15 minutes in StartingBlock T20 (PBS) Blocking Buffer (Thermo Fisher Scientific).

[0032] The primary antibody was replaced with a 1000-fold diluted antibody, and incubated at 4°C for two nights with shaking. The primary antibodies used were Anti-Rabbit-PIEZP2 Polyclonal antibody (ATLAS), Anti-Rabbit IGF1B Specific Polyclonal antibody (proteintech), Anti-Rabbit-Oxytocin-neurophysin1 antibody (abcam), and Anti-mouse β-actin antibody (Santacruz). The cells were then washed three times with 0.05% Tween20-PBS, and incubated at room temperature for 1 hour with HRP-Rabbit-IgG (Cell signaling) and HRP-Mouse-IgG (Invitrogen) diluted 1:10,000 in blocking buffer.

[0033] The cells were incubated with ECL Western blotting detection reagent (GE Healthcare) for 1 minute, and then detected and analyzed using a luminometer image analyzer LAS-4000 mini series (GE Healthcare). The protein amounts of piezo2, IGF-1, and oxytocin were calculated relative to β-actin.

[0034] (result) The expression levels were normalized to the control and the results are shown in FIGS. It was confirmed that piezo2 expression in Merkel cells was promoted by decapeptide-4 (0.0001% by weight), hydrolyzed yeast (0.000025% by weight), and palmitoyl tetrapeptide-7 (0.000025% by weight), which are peptides containing arginine as a constituent amino acid. It was also confirmed that IGF-1 expression in Merkel cells was promoted by decapeptide-4 (0.0001% by weight), hydrolyzed yeast (0.000025% by weight), and palmitoyl tetrapeptide-7 (0.000025% by weight), which are peptides containing arginine as a constituent amino acid. Furthermore, it was found that decapeptide-4 (0.0001% by weight), hydrolyzed yeast (0.000025% by weight), and palmitoyl tetrapeptide-7 (0.000025% by weight) promoted oxytocin expression. On the other hand, palmitoyl tetrapeptide-5, which does not contain arginine as a constituent amino acid, did not show any increase in piezo2, IGF-1, or oxytocin expression at any concentration.

Claims

1. A Merkel cell activity promoter containing decapeptide-4 or palmitoyl tetrapeptide-7 as an active ingredient.

2. A Merkel cell piezo2 expression promoter comprising decapeptide-4 or palmitoyl tetrapeptide-7 as an active ingredient.

3. An agent for promoting IGF-1 expression in Merkel cells, comprising decapeptide-4 or palmitoyl tetrapeptide-7 as an active ingredient.

4. An agent for promoting oxytocin expression in Merkel cells, comprising decapeptide-4 or palmitoyl tetrapeptide-7 as an active ingredient.

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