CN119816291A

CN119816291A - Cuticle formation promoter

Info

Publication number: CN119816291A
Application number: CN202380060990.5A
Authority: CN
Inventors: 中西忍; 筒井大气; 染井菜绪
Original assignee: Shiseido Co Ltd
Current assignee: Shiseido Co Ltd
Priority date: 2022-09-20
Filing date: 2023-05-15
Publication date: 2025-04-11
Also published as: JPWO2024062675A1; WO2024062675A1

Abstract

Provided are a stratum corneum formation promoter, a skin roughness inhibitor, a moisturizer, a skin barrier function improver, a melanin production inhibitor, and a screening method thereof that act via olfactory receptor activation. By using OR10A6 as an indicator, it is possible to screen a stratum corneum formation promoter, a skin roughness inhibitor, a moisturizer, a skin barrier function improver, and a melanin production inhibitor.

Description

Agent for promoting formation of horny layer

Technical Field

The present invention relates to a promoter for formation of a horny layer, a skin roughness inhibitor, a moisturizer, a skin barrier function improving agent, a melanogenesis inhibitor, and a screening method for such agents. In particular, the present invention relates to an agent for promoting formation of a horny layer, an agent for inhibiting skin roughness, a humectant, an agent for improving skin barrier function, and an agent for inhibiting melanin production, a screening method for such agents, an OR10A6 activator for promoting formation of a horny layer, inhibiting skin roughness, improving moisturizing function, skin barrier function, inhibiting melanin production, an OR5P2 activator, and an OR5P3 activator based on activation of OR10A6, OR5P2 OR5P3, and a composition comprising the same.

Background

Although olfactory receptors are mainly present in the nose and function as sensors for sensing odor, in recent years, it has been reported that some olfactory receptors are present not only in the nose but also in organs such as the skin, and have an ideal effect on the skin (non-patent document 1). For example, it has been reported that the olfactory receptor OR2AT4 is also present in keratinocytes, and if OR2AT4 in keratinocytes is activated by use of Sandalore which is an agonist of OR2AT4, wound healing and proliferation of keratinocytes are promoted, OR2AT4 is also present in sheath cells of hair follicles, and hair elongation is caused by use of Sandalore (non-patent documents 2, 3 and patent document 1).

However, more than 1,000 olfactory receptors exist depending on the species of mammal, and the presence of about 400 olfactory receptors that only act on humans has been confirmed. In order to smell a very wide variety of odors, these olfactory receptors react with not 1 but a plurality of substances to participate in a plurality of functions (non-patent document 4). For example, olfactory receptors OR6C74, OR51A7, OR4a15 expressed in sweat glands are reported to be involved in sweat regulation (patent document 2). Since the types and functions of the olfactory receptor are very large, there are many unknown substances that have an ideal effect on the skin by the presence and activation of the olfactory receptor, and further research is required.

Prior art literature

Patent literature

Patent document 1 Japanese patent application laid-open No. 2019-519502

Patent document 2 Japanese patent application laid-open No. 2021-127293

Non-patent literature

Non-patent literature 1:The American Physiological Society,June 13,2018;p.1739-1763, doi:10.1152/physrev.00013.2017

Non-patent documents 2:Experimental Dermatology,Volume 26,Issue 1,p.58-65, doi:10.1111/exd.13132

Non-patent literature 3:Nature FEBRUARY 2019,volume 18,p.116-138, doi:10.1038/s41573-018-0002-3

Non-patent document 4, i.e., package 15, no. 9 (2015), p401-406

Non-patent document 5:Pharmaceutics 2021,13,1314.https:// doi.org/10.3390/pharmaceutics13081314

Disclosure of Invention

Problems to be solved by the invention

The present inventors have conducted intensive studies on skin care for eyes and olfactory receptors, and have aimed at obtaining a promoter for formation of horny layer, a skin roughness inhibitor, a moisturizer, a skin barrier function improver, a melanogenesis inhibitor, and the like.

Means for solving the problems

The present inventors have found that OR10A6, OR5P2 and OR5P3, which are olfactory receptors, are expressed in the skin, and that an agonist of OR10A6, which is one of these olfactory receptors, enhances the activity of OR10A6 to exert an ideal effect on the skin such as promotion of horny layer formation, and completed the present invention.

The invention thus relates to the following:

[1]

A promoter for formation of horny layer contains OR10A6 activator as an active ingredient.

[2]

A promoter for forming horny layer comprises one or more selected from 3-phenylpropyl propionate, citronellol, phenethyl alcohol, alpha-cinnamyl alcohol, cyclamen aldehyde, geraniol, alpha-ionone, nerol, nonadecane, linalool, neomugwort aldehyde, phenylpropanol, androstadienone, androsterone and butyric acid as effective components.

[3]

A composition for promoting the formation of a horny layer, which comprises the horny layer formation promoter of [1] or [2 ].

[4]

A skin roughness inhibitor contains one or more selected from 3-phenylpropyl propionate, citronellol, phenethyl alcohol, alpha-cinnamyl alcohol, cyclamen aldehyde, geraniol, alpha-ionone, nerol, nonadecane, linalool, neoconvaldehyde, phenylpropanol, androstadienone, androsterone and butyric acid as an active ingredient.

[5]

A humectant contains 1 or more selected from 3-phenylpropyl propionate, citronellol, phenethyl alcohol, alpha-cinnamyl alcohol, cyclamen aldehyde, geraniol, alpha-ionone, nerol, nonadecane, linalool, neoconvaldehyde, phenylpropanol, androstenone, androsterone and butyric acid as effective components.

[6]

A skin barrier function improving agent contains 1 or more selected from 3-phenylpropyl propionate, citronellol, phenethyl alcohol, alpha-cinnamyl alcohol, cyclamen aldehyde, geraniol, alpha-ionone, nerol, nonadecane, linalool, neotame, phenylpropanol, androstadienone, androsterone and butyric acid as effective ingredients.

[7]

A melanogenesis inhibitor contains 1 or more selected from 3-phenylpropyl propionate, alpha-cinnamyl alcohol, cyclamen aldehyde, geraniol, alpha-ionone, nerol, nonadecane, linalool, neoconvalal, phenylpropanol, androstenone, androsterone and butyric acid as an active ingredient.

[8]

An OR10A6, OR5P2 OR5P3 activator for promoting formation of horny layer, inhibiting skin roughness, moisturizing, improving skin barrier function OR inhibiting melanogenesis.

[9]

A method for screening a promoter for formation of horny layer, a rough skin inhibitor, a moisturizer, a skin barrier function improver OR a melanogenesis inhibitor, wherein the activity of OR10A6, OR5P2 OR OR5P3 is used as an index.

[10]

The screening method according to [9], comprising:

Culturing the biological sample with a culture medium containing the candidate substance;

measuring the activity of OR10A6, OR5P2 OR OR5P3 in the biological sample, and

When the activity of the above-mentioned OR10A6, OR5P2 OR OR5P3 is increased as compared with the control, the candidate substance is determined to have the effect of promoting the formation of the horny layer, suppressing the roughness of the skin, moisturizing, improving the skin barrier function OR suppressing the production of melanin.

[11]

A kit for performing the screening method of any one of [9] OR [10], comprising reagents for determining the activity of OR10A6, OR5P2 OR OR5P 3.

[12]

A method for cosmetic purposes of promoting stratum corneum formation, inhibiting skin roughness, moisturizing, improving skin barrier function, OR inhibiting melanin production in a subject, comprising activating OR10A6, OR5P2, OR5P3 in the subject.

[13]

The method according to [12], wherein the OR10A6 is activated by administering the stratum corneum-forming promoter of [1], the skin roughness inhibitor of [4], the moisturizer of [5], the skin barrier function improver of [ 6), OR the melanogenesis inhibitor of [7] to a subject.

ADVANTAGEOUS EFFECTS OF INVENTION

By using 3-phenylpropyl propionate, androstadienone, androsterone, butyric acid OR an OR10A6 activator, it is possible to promote formation of a horny layer, inhibit skin roughness, improve moisturizing function, improve skin barrier function, and inhibit melanin production through activation of OR10 A6. Further, by evaluating the activity of OR10A6, the effect of promoting formation of a horny layer, suppressing skin roughness, moisturizing, improving skin barrier function, OR suppressing melanin production can be evaluated. By using such an evaluation method, it is possible to screen a promoter for formation of a horny layer, a skin roughness inhibitor, a moisturizer, a skin barrier function improver or a melanogenesis inhibitor.

Drawings

FIG. 1 shows the mRNA expression levels of OR10A6, OR5P2, OR5P3 and GAPDH as internal standards in keratinocytes.

FIG. 2 shows the concentration of Ca ²⁺ in keratinocytes as a fluorescence intensity ratio (F340/380). The horizontal axis of the graph on the left represents time (minutes), and the measurement start time is 0.0, which represents the change with time in the concentration of Ca ²⁺ when 3PPP was added at a time of about 0.5 minutes. Fig. 2 is a graph on the right side of the left graph, in which the change in the concentration of Ca ²⁺ due to the addition of 3PPP is expressed as a ratio of the maximum fluorescence intensity ratio (Δf340/380) with respect to the value at the measurement start time. Each point in the graph on the right represents the maximum fluorescence intensity ratio (Δf340/380) at regular intervals of time within 1.5 minutes from the measurement start time, and the solid line represents the average value thereof. The upper graph shows the results of adding 3PPP at each concentration (1.0 mM, 0.5mM, 0.05 mM), and the lower graph shows the results of comparing the increase in Ca ²⁺ concentration caused by the addition of 3PPP in OR10A6 knockdown keratinocytes (siOR A6: right) with control non-knockdown keratinocytes (siCont: left) in the case of adding 1.0mM 3 PPP. * By "P < 0.0005" is meant the statistically significant difference by ANOVA analysis of variance, scheffe method.

FIG. 3 shows the CE production rate (%) in keratinocytes to which 3PPP was added. The left panel shows the CE production (%) in non-knocked-down keratinocytes with 1.0mM added 3PPP (3 PPP+) or without (3 PPP-). The right panel shows the results of comparing the activity of OR10A6 in OR10A6 knockdown keratinocytes (siOR A6+) with OR without the addition of 1.0mM 3PPP (3 PPP+) with control non-knockdown keratinocytes (siOR 10A 6-). Experiments were performed with n=4, error bars mean ± SD, x mean statistically significant differences by t-test (left panel), ANOVA variance analysis, scheffe method (right panel) (< 0.05, P <0.001, P < 0.0001), ns mean no statistically significant differences.

Detailed Description

The present invention provides an accelerator for forming a horny layer, an inhibitor for rough skin, a humectant, an agent for improving skin barrier function, and an inhibitor for melanin production, which contain, as an active ingredient, 1 or more selected from the group consisting of 3-phenylpropyl propionate, citronellol, phenethyl alcohol, alpha-cinnamyl alcohol, cyclamen aldehyde, alpha-ionone, nerol, nonadecane, linalool, neoconyl aldehyde, phenylpropanol, androstenone, androsterone, and butyric acid.

3-Phenylpropyl propionate (CAS No. 122-74-7) has a floral fragrance and is used mainly as a perfume. In this specification, this is sometimes referred to as 3PPP.

Citronellol (CAS number: 106-22-9) is an aroma component contained in essential oils such as geranium essential oil and citronella essential oil, and has rose-like aroma.

Phenethyl alcohol (CAS No. 60-12-8) is contained in rose, wine, etc., and is used as a perfume in addition to the rose-like aroma, and is also used in stock, etc., by its antibacterial action.

Alpha-cinnamyl alcohol (CAS No. 4407-36-7) is also called cinnamyl alcohol or the like, and is contained in leaves of cinnamon or the like and has a hyacinth-like fragrance.

Cyclamal (CAS number: 103-95-7) is an artificial fragrance material that does not occur naturally, although it has a cyclamen-like, convallaria-like fragrance.

Geraniol (CAS number: 106-24-1) is contained in geranium essential oil, citronella essential oil, etc., and is known as a rose-like aroma component.

Alpha-ionone (CAS number: 127-41-3) is one of terpenes, and is 1 of isomers of alpha-ionone, beta-ionone and gamma-ionone. Each isomer has a different aroma and the alpha-ionone has an violet-like aroma.

Nerol (CAS number: 106-25-2) is a fragrance component contained in nerol, rose essential oil, etc., and having rose-like fragrance.

Nonadecane (CAS No. 629-92-5) is a component having a waxy odor contained in rose essential oil or the like.

Linalool (CAS number: 78-70-6) has (R) isomer and (S) isomer, the (S) isomer has orange-like fragrance, and the (R) isomer has lavender-like fragrance, and is used as a perfume, but is also used as a synthetic raw material for geraniol, citral and the like. Racemates can be preferably used.

New Convallaria (CAS number: 31906-04-4), also known as Convallaria, is a fragrance having an European-like fragrance.

Phenylpropanol (CAS No. 122-97-4) has a spice-like fragrance, and is used as a perfume, a stock, or the like.

Androstadienone (CAS number: 4075-07-4) is a steroid known to exhibit pheromone action, having a characteristic fragrance and sometimes formulated into perfumes.

Androsterone (CAS No. 453-41-8) is a male hormone which is also a causative agent of body odor, but is sometimes blended into a steroid in perfume due to its peculiar smell.

Butyric acid (CAS No. 107-92-6) is a naturally widely distributed substance having a characteristic unpleasant odor, but is sometimes contained in a perfume due to its characteristic odor.

3-Phenylpropyl propionate, citronellol, phenethyl alcohol, alpha-cinnamyl alcohol, cyclamen aldehyde, geraniol, alpha-ionone, nerol, nonadecane, linalool, neotame, phenylpropanol, androstadienone, androsterone and butyric acid are known to act as agonists of one of the olfactory receptors OR10 A6. Although these substances are known mainly as odor substances and pheromone substances, the inventors have found that they have an ideal skin-friendly effect such as promoting formation of a horny layer, suppressing skin roughness, moisturizing, improving skin barrier function and suppressing melanin production. In particular, such an action is exerted via activation of OR10 A6. These components may be synthesized using commercially available products.

Further, it is considered that even if the components other than the above are used, the OR10A6 can be activated, and the skin-friendly effects such as the effects of promoting the formation of the horny layer, suppressing the roughness of the skin, moisturizing, improving the skin barrier function and suppressing the generation of melanin can be exerted. Further, it was found that OR5P2 and OR5P3 are also expressed in keratinocytes in the same manner as OR10A6, and thus, by activating these olfactory receptors, it is expected to exert skin-friendly effects such as effects of promoting formation of the horny layer, suppressing skin roughness, moisturizing, improving skin barrier function and suppressing melanogenesis. Accordingly, the present invention also provides an OR10A6 activator, OR5P2 activator OR5P3 activator for promoting formation of the horny layer, inhibiting skin roughness, moisturizing, improving skin barrier function and inhibiting melanin production. Examples of the OR10A6 activator include an agent containing, as an active ingredient, a substance having an OR10A6 agonist action such as 3-phenylpropyl propionate, citronellol, phenethyl alcohol, α -cinnamyl alcohol, cyclamen aldehyde, geraniol, α -ionone, nerol, nonadecane, linalool, neoconvalal, phenylpropanol, androstenone, androsterone, and butyric acid. Among them, 3-phenylpropyl propionate, citronellol, and phenethyl alcohol are preferable, and 3-phenylpropyl propionate is particularly preferable. Examples of the OR5P3 activator include an agent containing a substance having an OR5P3 agonist action such as 1-octanol, acetophenone, coumarin, and the like as an active ingredient.

OR10A6, OR5P2 OR OR5P3 is one of the olfactory receptors and is a protein encoded by the OR10A6, OR5P2 OR OR5P3 gene, respectively. Olfactory receptors are present in cilia of olfactory cells in the nasal cavity, and if the ion channel of the cells is opened by the binding of odor molecules, sodium ions flow into the cells, action potential is generated, and odor information is transmitted to the brain, thereby causing perception of odor. Although several olfactory receptors have been reported to be present in tissues other than the nose as described above and to participate in actions other than the transmission of odor-free information, the inventors have found that OR10A6, OR5P2 OR5P3 are also present in the skin and that the activation of OR10A6 further exerts desirable actions on the skin such as actions of promoting formation of the horny layer, suppressing skin roughness, moisturizing, improving skin barrier function, and suppressing melanin production.

In one embodiment, the activation of OR10A6, OR5P2, OR5P3 means that, for example, if the OR10A6, OR5P2, OR5P3 activator of the present invention is added, the amount of mRNA OR protein of OR10A6, OR5P2, OR5P3 in the biological sample increases as compared with the case where the activator is not added. For example, an increase may refer to an increase having a statistically significant difference (e.g., student's t-test) with a significance level of 5%, and/or an increase of, for example, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more.

The measurement of the expression of OR10A6, OR5P2 OR OR5P3 can be determined by measuring the amount of mRNA OR the amount of protein of OR10A6, OR5P2 OR OR5P3 in a biological sample. The measurement of mRNA can be performed by using quantitative PCR, northern blotting, or other methods known in the art. Probes for mRNA of OR10A6, OR5P2 OR OR5P3, for example, can be used. Regarding the amount of protein, western blotting, immunostaining, ICM, and the like can be performed using methods known in the art.

In one embodiment, the activation of OR10A6, OR5P2, OR OR5P3 is caused by binding of 3-phenylpropyl propionate, citronellol, phenethyl alcohol, α -cinnamyl alcohol, cyclamen aldehyde, geraniol, α -ionone, nerol, nonadecane, linalool, neoconvalal, phenylpropanol, androstadienone, androsterone, butyric acid, 1-octanol, acetophenone, coumarin, other odor molecules, and substances having an activating effect on OR10A6, OR5P2, OR OR5P3 (e.g., agonists on the above olfactory receptors, etc.) to the olfactory receptors, respectively. Activation of OR10A6, OR5P2, OR5P3 can be measured, for example, by intracellular influx of cations such as Na ⁺、K⁺ OR Ca ²⁺ into the presence of these receptors in a biological sample. For example, if the cation such as intracellular Ca ²⁺ is increased as compared with before the addition if the candidate substance is added, it can be determined that the candidate substance has an OR10A6, OR5P2, OR5P3 activating effect. For example, when a candidate substance is added, the increase in cation concentration can mean, for example, an increase in a statistically significant difference (e.g., ANOVA variance analysis, scheffe method, etc.) in which the level of significance is set to 5%, or an increase of, for example, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more. The inflow of cations is not limited, but may be measured by intracellular imaging using a fluorescence microscope or the like.

In the case where the activation in the case where the candidate substance is added is increased as compared with the activation of the control OR10A6, OR5P2 OR5P3, the candidate substance may also be determined to be a substance having the activation of OR10A6, OR5P2 OR5P 3. Control OR10A6, OR5P2, OR5P3 activation can be determined by an experimental system to which a control agent is added, an experimental system under the same conditions except that only candidate substances are not included, OR an experimental system including OR10A6, OR5P2, OR5P3 knockdown cells via RNAi OR the like using shRNA, siRNA, OR knockdown cells via genome editing OR the like using TALEN, CRISPR.

The biological sample may be any sample containing cells that can be measured for the activity of OR10A6, OR5P2 OR5P3, and may contain cells other than keratinocytes. Examples of the cells other than keratinocytes include cells in which the presence of OR10A6, OR5P2, OR5P3 is confirmed by olfactory cells, cells produced so as to have OR10A6, OR5P2, OR5P3, and the like. The biological sample may be derived from any animal, but is preferably derived from a human from the viewpoints of cosmetic and pharmaceutical development. The biological sample may be a culture of cells capable of measuring the olfactory receptor, for example, keratinocytes OR other cells capable of measuring the activity of OR10A6, OR5P2 OR5P3, OR may be a skin sample, OR a cultured skin model constructed in 3 dimensions may be used.

The effects of promoting the formation of the horny layer, suppressing the roughness of the skin, moisturizing and improving the skin barrier function can be confirmed, for example, by measuring the cornified outer membrane (sometimes referred to as CE). For example, the amount of the produced keratinized outer film is obtained by lysing keratinocytes with a cell lysis solution containing an alcohol such as β -mercaptoethanol, a surfactant such as SDS, or the like, and counting the number of the keratinized outer films remaining undissolved in the solution. For example, in the case where the nerve activation effect in the case where the candidate substance is added is increased as compared with the control, the produced amount of the cornified envelope measured by such an operation can be determined that the candidate substance has the effects of promoting the formation of the horny layer, suppressing the skin roughness, moisturizing and improving the skin barrier function. For example, an increase in the amount of produced keratinized outer membrane can mean an increase in a statistically significant difference (e.g., t-test, dunnett test, etc.) of 5% in the level of significance, for example, or an increase of, for example, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more, as compared to a control, when the candidate substance is added. The amount of control produced of the cornified envelope can be determined by using an experimental system containing the control agent and an experimental system under the same conditions except that the candidate substance alone is not contained, OR by using an experimental system containing the OR10A6, OR5P2 OR5P3 knockdown cells and knockdown cells. The effects of promoting the formation of the horny layer, suppressing the roughness of the skin, moisturizing and improving the skin barrier function can also be confirmed by methods other than CE measurement, such as proliferation markers of keratinocytes, visual inspection, TEWL, measurement of specific proteins and lipids. For example, it can be confirmed by measuring the gene expression level and protein production level of TGM1 (Transglutaminase-1), FLG (Filaggrin), HAS3 (hyaluronate synthase3 (Hyaluronan Synthase 3)), GBA (β -glucocerebrosidase (β -Glucocerebrosidase)), IL-1α (Interleukin 1 Alpha)), and the like. The effect of inhibiting melanin production can be determined by measuring the amount of mRNA or the amount of protein of genes involved in melanin production such as IL-1. Alpha., COX2, etc. The measurement of mRNA can be performed by using quantitative PCR, northern blotting, or other methods known in the art. Probes for mRNA such as TGM1, FLG, HAS3, GBA, IL-1α, etc. can be used. Regarding the amount of protein, western blotting, immunostaining, ICM, and the like can be performed using methods known in the art. For example, promotion of horny layer formation, inhibition of skin roughness, moisturizing or improvement of skin barrier function can be confirmed by, for example, an increase in the amount of TGM1, FLG, HAS3, GBA mRNA or protein and/or a decrease in the amount of IL-1α mRNA or protein in a biological sample. For example, an increase can refer to an increase having a statistically significant difference (e.g., student's t-test, dunnett test) with a significance level of 5%, and/or an increase of, for example, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more. For example, a reduction can refer to a reduction having a statistically significant difference in the level of significance of 5% (e.g., student's t-test, dunnett test), and/or a reduction of, for example, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 100% or more.

The OR10A6, OR5P2, OR5P3 knockdown cell refers to, for example, a cell in which the expression of OR10A6, OR5P2, OR5P3 is impaired OR reduced by performing knockdown treatment on the OR10A6, OR5P2, OR5P3 gene. The knockdown treatment may be performed using any known technique such as RNAi using shRNA or siRNA. By the knock-down treatment, OR10A6, OR5P2, OR5P3 expression is impaired OR reduced, it may be meant, for example, that the amount of expression of the OR10A6, OR5P2, OR5P3 gene and/OR the amount of OR10A6, OR5P2, OR5P3 protein in a cell subjected to the knock-down treatment is reduced, for example, by a statistically significant difference (e.g., t-test, etc.) of 5% in significance level, OR by, for example, a reduction of 5% OR more, 10% OR more, 20% OR more, 30% OR more, 40% OR more, 50% OR more, 60% OR more, 70% OR more, 80% OR more, 90% OR 100% compared to a non-knockdown cell.

The OR10A6, OR5P2, OR5P3 knockout cell refers to a cell from which the function of the OR10A6, OR5P2, OR5P3 gene has been completely removed by, for example, performing a knockout treatment on the OR10A6, OR5P2, OR5P3 gene. The knock-out treatment may use any known technique such as genome editing using TALEN or CRISPR.

An embodiment of the present invention also provides a method for promoting formation of the stratum corneum, inhibiting skin roughness, moisturizing, improving skin barrier function, OR inhibiting melanin production in a subject, comprising applying a composition selected from the group consisting of 3-phenylpropyl propionate, citronellol, phenethyl alcohol, alpha-cinnamyl alcohol, cyclamen aldehyde, geraniol, alpha-ionone, nerol, nonadecane, linalool, neoconvalal, phenylpropanol, androstenone, androsterone, and butyric acid, OR10A6 activator, OR5P2 activator, OR5P3 activator (hereinafter sometimes collectively referred to as the agents of the present invention), OR a composition comprising the same. The invention also provides a method for promoting stratum corneum formation, inhibiting skin roughness, moisturizing, improving skin barrier function, OR inhibiting melanin production in a subject comprising activating OR10A6, OR5P2, OR5P3 in the subject. In one aspect of this embodiment, the activation of OR10A6, OR5P2, OR OR5P3 in a subject is achieved by the use of 3-phenylpropyl propionate, citronellol, phenethyl alcohol, alpha-cinnamyl alcohol, cyclamen aldehyde, geraniol, alpha-ionone, nerol, nonadecane, linalool, neoconvalal, phenylpropanol, androstenone, androsterone, butyric acid, 1-octanol, acetophenone, and coumarin, OR an agent of the invention, OR a composition comprising the same.

Examples of the subjects to which the method of the present invention is applied include subjects having reduced horny layer formation ability, subjects having OR desiring to prevent OR reduce rough skin, subjects having dry skin OR the like and desiring to moisturize OR desiring to moisturize, subjects having reduced skin barrier function OR desiring to improve skin barrier function, such as subjects having fear of sunburn, spots, dark subjects, subjects desiring to whiten, subjects desiring to inhibit melanin production, subjects having reduced OR5P2 OR5P3 activity, subjects having reduced OR olfactory receptor activity, such as subjects having reduced olfactory activity due to aging OR the like.

The method according to the present application is intended for cosmetic purposes, and medical actions performed by doctors and medical practitioners are sometimes excluded. The method according to the present application may be a method for supporting a cosmetic behavior of a subject.

A preferred embodiment is a cosmetic method for promoting formation of the stratum corneum, inhibiting skin roughness, moisturizing, improving skin barrier function, or inhibiting melanin production in a subject comprising applying 3-phenylpropyl propionate. In one aspect of such embodiments, promoting stratum corneum formation, inhibiting skin roughness, moisturizing, improving skin barrier function, OR inhibiting melanin production is achieved by activating OR10A6 of the subject.

Further the present invention also provides a composition comprising one or more selected from the group consisting of 3-phenylpropyl propionate, citronellol, phenethyl alcohol, alpha-cinnamyl alcohol, cyclamen aldehyde, geraniol, alpha-ionone, nerol, nonadecane, linalool, neoconvalsal, phenylpropanol, androstenone, androsterone, 1-octanol butyrate, acetophenone and coumarin, or an agent of the present invention, for promoting keratosis, inhibiting skin roughness, moisturizing, improving skin barrier function, and/or inhibiting melanin production.

The agent or composition of the present invention may be applied by any route including transdermally, orally, transmucosally, nasally, intravenously, intra-arterially, subcutaneously, etc., but is preferably applied transdermally from the viewpoint of acting on the skin. In the case of transdermal administration, it can be applied to any skin such as the skin of the face, head, neck, limbs, trunk.

The agent of the present invention may be blended into cosmetics, medicines or external products of medical department, respectively. These agents may be administered orally, or parenterally, such as transdermally. In the case of transdermal application, it can be formulated into skin external preparations. The external preparation for skin is not particularly limited as long as it can be applied to the skin, and any form such as solution, emulsion, solid, semisolid, powder dispersion, water-oil two-layer separation, water-oil-powder three-layer separation, ointment, gel, aerosol, wire, and stick can be used. In the case of being formulated into a skin external preparation, a base which is generally used in a skin external preparation, and an excipient such as a preservative, an emulsifier, a pH adjuster, and the like can be used. When blended in cosmetics, the composition can be blended in cosmetics for face or body, such as lotions, emulsions, lotions, creams, lotions, masks, essences, gels, make-up cosmetics for foundation, barrier creams, concealers, etc., bath agents, etc. By using a cosmetic comprising the agent of the present invention, activation, for example via OR10A6, OR5P2 OR5P3, results in promotion of stratum corneum formation, inhibition of skin roughness, moisturization, improvement of skin barrier function OR inhibition of melanin production. Therefore, the agent of the present invention can be used in cosmetics for the purpose of promoting formation of the horny layer, suppressing skin roughness, moisturizing, improving skin barrier function or suppressing melanin production.

The amount of the active ingredient in the agent or composition of the present invention may be arbitrarily selected from the viewpoint of exerting an effect of promoting formation of a horny layer, suppressing skin roughness, moisturizing, improving skin barrier function or suppressing melanogenesis. For example, from the viewpoint of blending as a skin external preparation, 3-phenylpropyl propionate, citronellol, phenethyl alcohol, α -cinnamyl alcohol, cyclamen aldehyde, geraniol, α -ionone, nerol, nonadecane, linalool, neoconvalal, phenylpropanol, androstenone, androsterone, butyric acid, 1-octanol, acetophenone, coumarin, OR other active ingredients capable of activating OR10A6, OR5P2 OR5P3 may be blended in an amount of 0.0005 to 100.0 mm. From the viewpoint of sufficiently exhibiting the effect, the amount may be preferably 0.05mM or more, and more preferably 0.5mM or more, for example, 1.0 mM. On the other hand, from the viewpoint of avoiding strong odor, the amount may be preferably 10.0mM or less, and more preferably 5.0mM or less. The above components may be combined in any ratio, and in this case, the total amount of these components is preferably within the above range.

The present invention also provides a method for screening a promoter for formation of a horny layer, a rough skin inhibitor, a moisturizer, a skin barrier function improver, OR a melanogenesis inhibitor, wherein the activity of OR10A6, OR5P2, OR OR5P3 is used as an index.

In one aspect, the screening method of the present invention comprises the steps of:

measuring the activity of OR10A6, OR5P2 OR OR5P3 in the biological sample, and

In the case where the activity of OR10A6, OR5P2 OR5P3 is increased as compared with the control, the candidate substance is determined to be a promoter for formation of a horny layer, a rough skin inhibitor, a moisturizing agent, a skin barrier function improving agent OR a melanogenesis inhibitor.

As the candidate substance, a substance belonging to any of a cosmetic raw material library, an extract library, a drug library, a compound library, and the like can be used. Examples thereof include, but are not limited to, odorous substances and perfume raw materials that react with olfactory receptors.

Further the present invention also provides an agonist for OR10A6, OR5P2 OR5P3 for promoting keratosis, inhibiting skin roughness, moisturizing, improving skin barrier function OR inhibiting melanin production, wherein preferably the promotion of keratosis, inhibiting skin roughness, moisturizing, improving skin barrier function OR inhibiting melanin production is achieved by activating OR10A6, OR5P2 OR5P3, such as 3-phenylpropyl propionate, citronellol, phenethyl alcohol, α -cinnamyl alcohol, cyclanilide, geraniol, α -ionone, nerol, nonadecane, linalool, phenylpropanol, androstenone, androsterone, butyric acid, 1-octanol, acetophenone OR coumarin. Preferred embodiments are 3-phenylpropyl propionate for promoting keratogenesis, inhibiting skin roughness, moisturizing, improving skin barrier function OR inhibiting melanin production, where the promotion of keratogenesis, inhibiting skin roughness, moisturizing, improving skin barrier function OR inhibiting melanin production is achieved by activating OR10 A6.

Further, the present invention also provides a use of 3-phenylpropyl propionate, citronellol, phenethyl alcohol, α -cinnamyl alcohol, cyclamen aldehyde, geraniol, α -ionone, nerol, nonadecane, linalool, neoconvalsal, phenylpropanol, androstenone, androsterone, butyric acid and other agonists against OR10A6, OR agonists against OR5P2 OR agonists against OR5P3 such as 1-octanol, acetophenone, coumarin and other agonists for the manufacture of a medicament for promoting horny layer formation, inhibiting skin roughness, moisturizing, improving skin barrier function OR inhibiting melanin production.

All documents mentioned in this specification are incorporated by reference in their entirety into this specification.

The following examples of the present invention are given for illustrative purposes only and are not intended to limit the technical scope of the present invention. The technical scope of the present invention is limited only by the description of the claims. The modification of the present invention, for example, addition, deletion, and substitution of the constituent elements of the present invention may be performed without departing from the spirit of the present invention.

Example 1

Example 1 expression of OR10A6, OR5P2 and OR5P3 in keratinocytes

Keratinocytes (vendor: kun-Kyoku) were cultured in EPILIFE-KG2 medium (vendor: kun-Kyowa) under a humidified atmosphere at 37℃to collect a confluent culture, and cDNA was prepared using Trizol (vendor: NIPPON GENE) according to the product instructions of VILO Master Mix (vendor: invitrogen). Real-time PCR was performed using the primers for OR10A6 amplification (SEQ ID NO: 1, 2), OR5P2 amplification (SEQ ID NO: 3, 4), OR5P3 amplification (SEQ ID NO: 5, 6) and internal standard GAPDH amplification (SEQ ID NO: 7, 8) of the sequences shown below, and the amplification products were confirmed by electrophoresis.

OR10A6 Forward 5'-tatttacaacccaaatctg-3' (SEQ ID NO. 1)

OR10A6 reverse 5'-tcagattgtgtgtaaaacc-3' (SEQ ID NO. 2)

OR5P2 Forward 5'-accttcatttatgtgatgc-3' (SEQ ID NO. 3)

OR5P2 reverse 5'-aaaataacaagcatcatgag-3' (SEQ ID NO. 4)

OR5P3 Forward 5'-cagtcactctgttctatgg-3' (SEQ ID NO. 5)

OR5P3 reverse 5'-taagctctctcttcagagc-3' (SEQ ID NO. 6)

GAPDH Forward 5'-gaaggtgaaggtcggagtc-3' (SEQ ID NO: 7)

GAPDH reverse 5'-gaagattggtgatgggatttc-3' (SEQ ID NO: 8)

The results are shown in fig. 1. It was confirmed that OR10A6, OR5P2 and OR5P3 were all expressed in keratinocytes. Thus, the response in keratinocytes was investigated by the following experiment using OR10A6 as one of these olfactory receptors and 3PPP as an agonist thereof.

Example 2 calcium response observations of cells Using 3PPP

Keratinocytes (vendor: coku) were cultured in EPILIFW-KG2 medium (vendor: coku) OR EPILIFW-KG2 medium containing OR10A6 siRNA (vendor: horizon) for 1 day under a humidified atmosphere at 37 ℃. For cells that became confluent, the cells were replaced with EPILIFW-KG2 containing Fura2 AM (vendor: life technology) at 5. Mu.M and cultured for 1 hour. Then, the cells were replaced with a buffer (pH 7.4) containing 150mM NaCl, 5mM KCl, ₂ 1.8mM、MgCl₂ 1.2.2 mM CaCl, 25mM HEPES, and 10mM D-glucose, and 3-phenylpropyl propionate (3 PPP) (vendor: sigmaAldrich) was added at a concentration of 0.05 to 1 mM. The fluorescence intensity ratio (F340/380) was measured as the change in intracellular Ca ²⁺ concentration at each time when 3PPP was added by observing the wavelengths of 340nm and 380nm with a fluorescence microscope (vendor: コ N).

The results are shown in fig. 2. As shown in the above graph, if 3PPP is added, the Ca ²⁺ concentration increases significantly from the value before the addition of 3 PPP. Further, it was found that the increase in the Ca ²⁺ concentration significantly increased as the concentration of 3PPP increased to 0.05mM, 0.5mM, and 1.0mM, depending on the concentration. Further, it was confirmed from the following graph that the reaction using 3PPP was significantly lower in OR10A6 knockdown keratinocytes. From the above experiments, it was found that OR10A6 present in keratinocytes was activated by 3PPP as an agonist thereof. Therefore, the effect on the skin caused by activating OR10A6 by 3PPP was investigated by the following experiment.

Example 3 effects of OR10A6 activation on promotion of stratum corneum formation, inhibition of skin roughness, moisture retention and improvement of skin Barrier function 1

Keratinocytes (vendor: coku) were cultured in EPILIFW-KG2 medium (vendor: coku) OR EPILIFW-KG2 medium containing OR10A6 siRNA (vendor: horizon) for 1 day under a humidified atmosphere at 37 ℃. For cells that became confluent, 3-phenylpropyl propionate (3 PPP) (vendor: sigmaAldrich) was added to the cells at a concentration of 1mM and further cultured for 3 days. Cultured cells were collected by trypsin (vendor: nap and ku corporation) treatment after 3 days, and the total number of cells was counted by an optical microscope, and the number of dead cells stained with trypan blue (vendor: nap and ku corporation) was further counted, whereby the number of living cells was obtained as the total number of cells-dead cells. After centrifugation, the cells were replaced with a buffer solution containing 1% of β -mercaptoethanol, 1% of SDS (sodium lauryl sulfate) and 20mM of trichcl (pH 7.5), and the cells were lysed at 95℃for 15 minutes, and the number of keratinocytes having a keratinized outer membrane remaining undissolved in the solution was counted by an optical microscope. As an index of the effect of promoting the formation of the horny layer, suppressing the roughness of the skin, moisturizing, and improving the skin barrier function, the amount of produced cornified outer membrane was calculated as the number of cornified outer membrane-containing keratinocytes/the number of living cells×100, and this value was expressed as CE production rate (%).

The results are shown in fig. 3. CE production in keratinocytes was significantly increased by 3PPP (left panel). However, no significant increase in the amount of keratinocytes produced by the keratinized outer film was observed even with the addition of 3PPP for the OR10A6 knockout keratinocytes. Further, without the addition of 3PPP, CE production was significantly reduced in OR10A6 knockout keratinocytes compared to OR10A6 non-knockout keratinocytes (right panel). Thus, it is suggested that CE generation is promoted by activation of OR10 A6.

From the above results, it was found that OR10A6, OR5P2 and OR5P3 were present in keratinocytes, and CE production was promoted by activation of OR10A6 by 3 PPP.

Example 4 effects of OR10A6 activation on promotion of stratum corneum formation, inhibition of skin roughness, moisture retention and improvement of skin Barrier function 2

The possibility that 3PPP promotes CE formation through activation of an olfactory receptor such as OR10A6 was shown in example 3, in which the expression of TGM1 gene encoding transglutaminase 1, an enzyme promoting CE formation and maturation through activation of OR10A6, was measured.

4-1 Cultivation of cells

HaCaT cells were seeded in 24-well plates (Cat No.3526, corning, USA) at a density of 10.0X10 ⁴ cells/well in Dulbecco's modified Eagle's Medium (Dulbecco's Modified Eagle Medium) (DMEM, cat No.043-30085, wako, japan) containing 10.0% (v/v) fetal bovine serum (Fetal Bovine Serum) (FBS, cat No. SH30071.03, hyclone, UK) and 1.0% (v/v) antifungal agent (Antibiotic-Antimycotic 100X,Cat No.15240-062, invitrogen, USA) and incubated in a CO ₂ incubator (CO ₂ concentration 5%,37 ℃) for 24 hours. After the removal of the medium, the medium was changed to a medium to which 3PPP was added so that the final concentration became 0.05mM, and further cultured in a CO ₂ incubator for 48 hours. The control used medium that did not contain 3 PPP.

4-2 RNA extraction/purification, quantification and purity determination

RNA extraction/purification was performed using PureLinkTM RNA MINI KIT (Cat No.12183018A, invitrogen, USA), and a portion of the purified RNA was collected into a UV-permeable 96-well plate, diluted 10-fold with Tris-EDTA Buffer, and absorbance at 230nm, 260nm, and 280nm (OD 230, OD260, OD 280) was measured using a microplate reader (SPARK (registered trademark) 10M TECAN,Switzerland). The RNA concentration was calculated using OD260 and diluted with TE buffer to adjust the RNA concentration to 10. Mu.g/mL.

4-3 Gene expression analysis Using real-time PCR method

Reverse transcription of RNA was performed using SuperScript (trade mark) IV VILOTM Master Mix with ezDNase (Cat No.11766050, invitrogen, USA). mu.L of SuperScript (trade name) IV VILOTM Master Mix and 6. Mu.L of Nuclease-free water (Nuclease-FREE WATER) were added to each 1 well of the 8-well tube, and the mixture was heated at 25℃for 10 minutes, 50℃for 10 minutes, and 85℃for 5 minutes using real-time PCR (QuantStudio (registered trade name) 3,Applied Biosystems,USA) to synthesize cDNA. mu.L of TaqMan (registered trademark) FAST ADVANCEDMASTER Mix (Cat No.4444557, applied Biosystems, USA), 1. Mu.L of TAQMAN GENE Expressior, 7. Mu.L of UltraPureTM DISTILLED WATER (Invitrogen, cat No.10977-015, USA) and 2. Mu.L of cDNA were added to each 1 well of the PCR plate, and sealed. Real-time qPCR was performed using a primer (Hs 00165929 _m1) for TGM1 and a primer (Hs 02786624 _g1) for GAPDH as an internal standard gene, and after calculating the Threshold Cycle (Ct) value of TGM1 in the 3PPP addition medium, the Ct value was corrected by GAPDH to be a ΔCt value, and assuming that the gene amount per 1 Cycle became 2-fold, the gene expression amount in the 3PPP addition medium was obtained when the gene expression amount of the control was 1. In the analysis of gene expression level, an average value of 24 well plates×3 wells was used for 1 treatment group.

The results are shown in table 1 below.

TABLE 1 promotion of TGM1 expression by 3PPP addition

(In the paired sample T test (both sides), the ratio of the non-added group to the non-added group was 1)

As shown in Table 1, if 3PPP was added, the gene expression of TGM1 was significantly increased, and 3PPP activated OR10A6 to promote the production of TGM1 and thus the formation and maturation of CE, suggesting the possibility of helping to promote the formation of horny layer, inhibit skin roughness, moisturize and improve skin barrier function.

Example 5 effects of OR10A6 activation on promoting stratum corneum formation, inhibiting skin roughness, moisturizing, and improving skin Barrier function 3

In order to confirm the effects of forming a horny layer, suppressing skin roughness, moisturizing, and improving skin barrier by the activation of OR10A6, the following experiments were performed using other genes involved in these functions. More specifically, expression amounts of FLG (Filaggrin) encoding Filaggrin involved in the moisture retention function of skin, HAS3 (hyaluronate synthase 3 (Hyaluronan Synthase 3)) encoding a hyaluronate synthase produced in epidermal cells and involved in the moisture retention of skin, and GBA (β -enzyme glucocerebrosidase (β -Glucocerebrosidase)) encoding a ceramide EOP (ceramide 1) involved in the barrier function of skin were measured as NMF.

The expression levels of these genes in the medium containing or not containing 0.05mM 3PPP were measured using the same materials and methods as in example 4, except that FLG (Hs 00856927 _g1), human hyaluronate synthase 3 (Human Hyaluronan Synthase) (HAS 3, assay ID. Hs00193436_m1), and the primers for Human beta-enzyme glucocerebrosidase (Human beta-Glucocerebrosidase) (GBA, assay ID. Hs00986836_g1) were used instead of the primer for TGM1 (Hs 00165929 _m1).

The results are shown in tables 2 to 4 below.

TABLE 2 promotion of FLG expression by 3PPP addition

TABLE 3 promotion of HAS3 expression by 3PPP addition

TABLE 4 promotion of GBA expression by 3PPP addition

As shown in tables 2 to 4, when 3PPP was added, the gene expression of FLG, HAS3 and GBA significantly increased. From the above results, it was suggested that OR10A6 activation using 3PPP OR the like promotes expression of these genes in addition to TGM1, thereby contributing to promotion of horny layer formation, inhibition of skin roughness, moisture retention and improvement of skin barrier function.

Example 6 effects of OR10A6 on promoting keratogenesis, inhibiting skin roughness, moisturizing and improving skin Barrier function 4

In order to examine whether OR10A6 activators other than 3PPP have the same effect, citronellol and phenethyl alcohol known as substances reacting with OR10A6 were used (non-patent document 5), and whether OR not these substances have the same keratolytic action as 3PPP was examined. More specifically, TGM1 gene expression was measured by the same materials and methods as in example 4, except that citronellol (CAS No. 106-22-9, and light 037-05993) or phenethyl alcohol (CAS No. 60-12-8, TCI P0084) was used in place of 3PPP at the following concentrations.

The results are shown in tables 5 and 6 below.

TABLE 5 promotion of TGM1 expression by citronellol addition

TABLE 6 promotion of TGM1 expression by phenethyl alcohol addition

According to the results of Table 6, TGM1 expression was also significantly increased using not only 3PPP but also other citronellol, phenethyl alcohol, as an OR10A6 activator.

Example 7 effects on promoting stratum corneum formation, inhibiting skin roughness, moisturizing, and improving skin Barrier function by OR10A6 activation 5

The following experiment was performed using GBA in the same manner as in example 5 with respect to citronellol and phenethyl alcohol, the effect of which was observed in example 6. More specifically, the GBA gene expression level was measured by the same materials and methods as in example 5, except that citronellol (CAS No. 106-22-9, and light 037-05993) or phenethyl alcohol (CAS No. 60-12-8, TCI P0084) was used in place of 3PPP at the following concentrations.

The results are shown in tables 7 and 8 below.

TABLE 7 promotion of GBA expression by citronellol addition

TABLE 8 promotion of GBA expression by phenethyl alcohol addition

According to the results of table 7, the expression of GBA was significantly OR significantly increased in a significant inclination even with citronellol and phenethyl alcohol as OR10A6 activators other than 3PPP in the same manner as in example 6.

Example 8 effects on promoting stratum corneum formation, inhibiting skin roughness, moisturizing, and improving skin Barrier function by OR10A6 activation 5

The following experiment was performed using HAS3 in the same manner as in example 5 with respect to citronellol having the effect observed in examples 6 and 7. More specifically, the HAS3 gene expression level was measured by the same materials and methods as in example 5, except that citronellol (CAS No. 106-22-9, and light 037-05993) was used in place of 3PPP at the following concentrations.

The results are shown in table 9 below.

TABLE 9 promotion of HAS3 expression by citronellol addition

Example 9:

From the above, it was suggested that OR10A6 was activated to exert the effects of promoting formation of horny layer, suppressing skin roughness, moisturizing and improving skin barrier function, but in order to confirm other effects, the expression amount of IL-1α (Interleukin-1α (intelukin 1 Alpha)) encoding IL- α known as inflammatory cytokine and promoting melanin production was measured. More specifically, the gene expression level of IL-1α in a medium containing or not containing 3PPP at the following concentration was measured by the same materials and methods as in example 4, except that the primer for IL-1α (Hs 00174092 _m1) was used instead of the primer for TGM1 (Hs 00165929 _m1). IL-1α (Interleukin 1 Alpha)) is known as a gene encoding IL-1α protein as an inflammatory cytokine, and IL-1α causes excessive production of melanin by increasing tyrosinase activity.

The results are shown in table 10 below.

TABLE 10 inhibition of IL-1 alpha expression by 3PPP addition

(Dunnett test, described in terms of the ratio of 1 for the non-added group)

As shown in Table 10, the gene expression concentration of IL- α was decreased in a dependent manner when 3PPP was added. According to the results, it was suggested that the activation of OR10A6 not only contributes to promotion of formation of horny layer, inhibition of skin roughness, moisture retention and improvement of skin barrier function by promotion of expression of TGM1, FLG, HAS3, GBA, but also contributes to inhibition of skin roughness, inhibition of melanin production possibility by inhibition of expression of IL- α.

The above results can be expected to bring about effects on skin that are desirable, such as effects of promoting formation of horny layer, suppressing skin roughness, moisturizing, improving skin barrier function, and suppressing melanin production, by activation of OR10A6, OR5P2, OR5P3, and these effects are achieved by components that activate OR10A6, OR5P2, OR5P 3. For example, 3-phenylpropyl propionate, citronellol, phenethyl alcohol, α -cinnamyl alcohol, cyclamen aldehyde, geraniol, α -ionone, nerol, nonadecane, linalool, neolily-of the valley aldehyde, phenylpropanol, androstenone, androsterone, butyric acid are known as components for activating OR10A6, and 1-octanol, acetophenone, coumarin, and the like are known as components for activating OR5P3, and therefore effects derived from these components can be expected.

Claims

1. A stratum corneum formation promoter comprising an OR10A6 activator as an active ingredient.

2. A stratum corneum formation promoter, comprising one or more selected from the group consisting of 3-phenylpropyl propionate, citronellol, phenylethyl alcohol, α-cinnamyl alcohol, cyclamen aldehyde, geraniol, α-ionone, nerol, nonadecane, linalool, lyral, phenylpropanol, androstenone, androsterone and butyric acid as active ingredients.

3. A composition for promoting stratum corneum formation, comprising the stratum corneum formation promoter according to claim 1 or 2.

4. A skin roughness inhibitor comprising one or more selected from the group consisting of 3-phenylpropyl propionate, citronellol, phenylethyl alcohol, α-cinnamyl alcohol, cyclamen aldehyde, geraniol, α-ionone, nerol, nonadecane, linalool, lyral, phenylpropanol, androstenone, androsterone and butyric acid as active ingredients.

5. A moisturizer comprising one or more selected from the group consisting of 3-phenylpropyl propionate, citronellol, phenylethyl alcohol, α-cinnamyl alcohol, cyclamen aldehyde, geraniol, α-ionone, nerol, nonadecane, linalool, lyral, phenylpropanol, androstenone, androsterone and butyric acid as active ingredients.

6. A skin barrier function improver comprising one or more selected from the group consisting of 3-phenylpropyl propionate, citronellol, phenylethyl alcohol, α-cinnamyl alcohol, cyclamen aldehyde, geraniol, α-ionone, nerol, nonadecane, linalool, lyral, phenylpropanol, androstenone, androsterone and butyric acid as active ingredients.

7. A melanin production inhibitor comprising one or more selected from the group consisting of 3-phenylpropyl propionate, α-cinnamyl alcohol, cyclamen aldehyde, geraniol, α-ionone, nonadecane, linalool, lyral, phenylpropanol, androstenone, androsterone and butyric acid as active ingredients.

8. An OR10A6 activator for promoting stratum corneum formation, inhibiting skin roughness, moisturizing, improving skin barrier function or inhibiting melanin production.

9. A method for screening a stratum corneum formation promoter, a skin roughness inhibitor, a moisturizer, a skin barrier function improver, or a melanin production inhibitor, wherein OR10A6 activity is used as an indicator.