Accelerator for formation of stratum corneum

Abstract

Provided are a cuticle formation promoter, a skin roughness inhibitor, a humectant, a skin barrier function improving agent, and a melanin production inhibitor which act via the activation of olfactory receptors, and methods for screening the same. By using OR10A6 as an index, screening of a cuticle formation promoter, a skin roughness inhibitor, a moisturizer, a skin barrier function improving agent, and a melanin production inhibitor can be performed.

Description

Technical Field

[0001] The present invention relates to a stratum corneum formation promoter, a skin roughness inhibitor, a moisturizer, a skin barrier function improver, and a melanin production inhibitor, and a method for screening such agents. In particular, the present invention relates to a stratum corneum formation promoter, a skin roughness inhibitor, a moisturizer, a skin barrier function improver, and a melanin production inhibitor based on the activation of OR10A6, OR5P2 or OR5P3, a method for screening such agents, an OR10A6 activator, an OR5P2 activator, and an OR5P3 activator for promoting stratum corneum formation, inhibiting skin roughness, improving moisturizing function, skin barrier function, and inhibiting melanin production, and a composition containing them. Background Art

[0002] Olfactory receptors are mainly present in the nose and function as sensors for perceiving odors, but in recent years, it has been reported that certain olfactory receptors are not only present in the nose, but also in organs such as the skin, bringing desirable effects to the skin (Non-Patent Document 1). It has also been reported that, for example, the olfactory receptor OR2AT4 is also present in keratinocytes, and if OR2AT4 in keratinocytes is activated by the application of Sandalore, which is an agonist of OR2AT4, wound healing and proliferation of keratinocytes are promoted; OR2AT4 is also present in the sheath cells of hair follicles, and the application of Sandalore causes hair elongation (Non-Patent Documents 2, 3, Patent Document 1).

[0003] However, there are more than 1,000 kinds of olfactory receptors depending on the type of mammals, and for humans, only about 400 kinds of functional olfactory receptors have been confirmed. Moreover, in order to be able to smell a very diverse variety of odors, these olfactory receptors react with not one but multiple substances and participate in multiple functions (Non-patent Document 4). For example, it is reported that the olfactory receptors OR6C74, OR51A7, and OR4A15 expressed in sweat glands are involved in sweat regulation (Patent Document 2). In this way, since there are many types and functions of olfactory receptors, there are many unknown parts about which olfactory receptors exist in the skin and are activated by them to bring about a desired effect on the skin, and further exploration is required.

[0004] Prior art literature

[0005] Patent Literature

[0006] Patent Document 1: Japanese Patent Application No. 2019-519502

[0007] Patent Document 2: Japanese Patent Application Publication No. 2021-127293

[0008] Non-patent literature

[0009] Non-patent literature 1: The American Physiological Society, June 13, 2018; p.1739-1763; doi:10.1152 / physrev.00013.2017

[0010] Non-patent literature 2: Experimental Dermatology, Volume 26, Issue 1, p.58-65; doi:10.1111 / exd.13132

[0011] Non-patent literature 3: Nature FEBRUARY 2019, volume 18, p.116-138; doi:10.1038 / s41573-018-0002-3

[0012] Non-patent document 4: Oreos, Vol. 15, No. 9 (2015), p401-406

[0013] Non-patent literature 5: Pharmaceutics 2021, 13, 1314. https: / / doi.org / 10.3390 / pharmaceutics13081314 Summary of the invention

[0014] Problems to be solved by the invention

[0015] The present inventors have conducted intensive studies on skin care focusing on olfactory receptors, with the aim of obtaining a stratum corneum formation promoter, a skin roughness inhibitor, a moisturizer, a skin barrier function improver, a melanin production inhibitor, and the like.

[0016] Means for solving problems

[0017] 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, one of these olfactory receptors, increases the activity of OR10A6, thereby exerting desirable effects on the skin such as promoting stratum corneum formation, thereby completing the present invention.

[0018] Therefore the present invention relates to the following: [1]

[0020] A stratum corneum formation promoter containing an OR10A6 activator as an active ingredient. [2]

[0022] A stratum corneum formation promoter contains one or more selected from 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]

[0024] A composition for promoting stratum corneum formation, comprising the stratum corneum formation promoter of [1] or [2]. [4]

[0026] 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]

[0028] A moisturizer contains 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]

[0030] A skin barrier function improver, comprising one or more selected from 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]

[0032] A melanin production inhibitor contains one or more selected from the group consisting of 3-phenylpropyl propionate, α-cinnamyl alcohol, cyclamen aldehyde, geraniol, α-ionone, nerol, nonadecane, linalool, lyral, phenylpropanol, androstenone, androsterone and butyric acid as active ingredients. [8]

[0034] An OR10A6, OR5P2 or OR5P3 activator for promoting stratum corneum formation, inhibiting skin roughness, moisturizing, improving skin barrier function or inhibiting melanin production. [9]

[0036] 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 the activity of OR10A6, OR5P2, or OR5P3 is used as an indicator.

[10]

[0038] The screening method according to [9] comprises:

[0039] culturing the biological sample with a culture medium containing the candidate substance;

[0040] measuring the activity of OR10A6, OR5P2 or OR5P3 in a biological sample; and

[0041] When the activity of OR10A6, OR5P2 or OR5P3 is increased compared to the control, the candidate substance is determined to have the effects of promoting stratum corneum formation, inhibiting skin roughness, moisturizing, improving skin barrier function or inhibiting melanin production.

[11]

[0043] A kit for carrying out the screening method described in any one of [9] or

[10] , comprising a reagent for determining the activity of OR10A6, OR5P2 or OR5P3.

[12]

[0045] A method for the cosmetic purpose of promoting stratum corneum formation, inhibiting skin roughness, moisturizing, improving skin barrier function or inhibiting melanin production in a subject, comprising: activating OR10A6, OR5P2 or OR5P3 in the subject.

[13]

[0047] According to the method described in

[12] , the activation of OR10A6 is achieved by administering the stratum corneum formation promoter described in [1], the skin roughness inhibitor described in [4], the moisturizer described in [5], the skin barrier function improving agent described in [6] or the melanin production inhibitor described in [7] to a subject.

[0048] Effects of the Invention

[0049] By applying 3-phenylpropyl propionate, androstenone, androsterone, butyric acid or an OR10A6 activator, the stratum corneum formation can be promoted, the skin roughness can be inhibited, the moisturizing function can be improved, the skin barrier function can be improved, and the melanin production can be inhibited through the activation of OR10A6. Further, by evaluating the activity of OR10A6, the effects of promoting stratum corneum formation, inhibiting skin roughness, moisturizing, improving skin barrier function or inhibiting melanin production can be evaluated. Using such an evaluation method, a stratum corneum formation promoter, a skin roughness inhibitor, a moisturizer, a skin barrier function improver or a melanin production inhibitor can be screened. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] Figure 1 The mRNA expression levels of OR10A6, OR5P2, OR5P3 and GAPDH as an internal standard in keratinocytes are shown.

[0051] Figure 2Left keratinocyte Ca 2+ The concentration is expressed as the fluorescence intensity ratio (F340 / 380). The horizontal axis of the left graph is time (minutes), with the measurement start time set to 0.0, indicating the Ca concentration when 3PPP was added at about 0.5 minutes. 2+ Changes in concentration over time. Figure 2 Right is a graph showing the Ca2+ induced by 3PPP addition for the left graph. 2+ The change in concentration is expressed as the ratio of the maximum fluorescence intensity ratio (ΔF340 / 380) relative to the value at the start of the measurement. Each point in the coordinate graph on the right represents the maximum fluorescence intensity ratio (ΔF340 / 380) at a certain time interval within 1.5 minutes from the start of the measurement, and the solid line represents its average value. The upper figure shows the results of adding 3PPP at various concentrations (1.0mM, 0.5mM, 0.05mM), and the lower figure shows the Ca2+ in OR10A6 knockdown keratinocytes (siOR10A6: right) with the addition of 1.0mM 3PPP. 2+ The results of the comparison of the increase in concentration with the control non-knockdown keratinocytes (siCont: left). ***, *** indicate statistically significant differences by ANOVA analysis of variance and Scheffe's method (***P<0.0005, ***P<0.0001).

[0052] Figure 3 Represents the CE production rate (%) in keratinocytes with the addition of 3PPP. The left figure represents the CE production rate (%) in non-knockdown keratinocytes with the addition of 1.0mM 3PPP (3PPP+) or without the addition of (3PPP-). The right figure is the result of comparing the OR10A6 activity in OR10A6 knockdown keratinocytes (siOR10A6+) with the control non-knockdown keratinocytes (siOR10A6-) with the addition of 1.0mM 3PPP (3PPP+) or without the addition of (3PPP-). The experiment was carried out with n=4, and the error bars represent the mean ± SD, * represents statistically significant differences (*P < 0.05, **P < 0.001, **P < 0.0001) by t-test (left figure), ANOVA analysis of variance, Scheffe method (right figure), and ns represents no statistically significant differences. DETAILED DESCRIPTION

[0053] The present invention provides a stratum corneum formation promoter, a skin roughness inhibitor, a moisturizer, a skin barrier function improver, and a melanin production inhibitor, which contain one or more selected from 3-phenylpropyl propionate, citronellol, phenylethyl alcohol, α-cinnamyl alcohol, cyclamen aldehyde, α-ionone, nerol, nonadecane, linalool, lyral, phenylpropanol, androstadienone, androsterone and butyric acid as active ingredients.

[0054] 3-Phenylpropyl propionate (CAS No.: 122-74-7) has a floral aroma and is mainly used as a fragrance. In this specification, it may be referred to as 3PPP.

[0055] Citronellol (CAS No.: 106-22-9) is an aroma component contained in essential oils such as geranium essential oil and citronella essential oil, and has a rose-like aroma.

[0056] Phenylethanol (CAS No.: 60-12-8) is contained in roses, wine, etc., and is used as a fragrance due to its rose-like aroma. In addition, it is used in preservatives, etc. due to its antibacterial effect.

[0057] α-Cinnamyl alcohol (CAS No.: 4407-36-7), also known as cinnamyl alcohol, is contained in cinnamon leaves and the like and has a hyacinth-like aroma.

[0058] Cyclamen aldehyde (CAS No.: 103-95-7) is an artificial flavoring substance that has a cyclamen or lily of the valley-like aroma but does not exist naturally.

[0059] Geraniol (CAS No.: 106-24-1) is contained in geranium essential oil, palmarosa essential oil, citronella essential oil, and the like, and is known as a rose-like aroma component.

[0060] α-ionone (CAS No.: 127-41-3) is a type of terpenoid and one of the isomers of α-ionone, β-ionone, and γ-ionone. Each isomer has a different aroma, and α-ionone has a violet-like aroma.

[0061] Nerol (CAS No.: 106-25-2) is an aroma component contained in orange blossom essential oil, rose essential oil, etc., and has a rose-like aroma.

[0062] Nonadecane (CAS No.: 629-92-5) is a component contained in rose essential oil and the like, and has a waxy odor.

[0063] Linalool (CAS No.: 78-70-6) exists in (R) isomers and (S) isomers. The (S) isomer has an orange-like aroma, and the (R) isomer has a lavender-like aroma. It is used as a fragrance, but is also used as a synthetic raw material for geraniol, citral, etc. The racemic form can be preferably used.

[0064] Lyral (CAS No.: 31906-04-4), also known as lily of the valley aldehyde, is a fragrance having a lily of the valley-like aroma.

[0065] Phenylpropanol (CAS No.: 122-97-4) has a spice-like aroma and is used as a fragrance, preservative, etc.

[0066] Androstadienone (CAS No.: 4075-07-4) is a steroid known to exhibit pheromone effects, has a unique aroma, and is sometimes formulated into perfumes.

[0067] Androstenone (CAS No.: 453-41-8) is a steroid that is a male hormone that causes body odor, but is sometimes blended into perfumes due to its unique odor.

[0068] Butyric acid (CAS No.: 107-92-6) is a substance that has a unique unpleasant odor and is widely distributed in nature. However, due to its unique odor, it is sometimes included in fragrances.

[0069] 3-Phenylpropyl propionate, citronellol, phenylethyl alcohol, α-cinnamyl alcohol, cyclamen aldehyde, geraniol, α-ionone, nerol, nonadecane, linalool, lyral, phenylpropanol, androstadienone, androsterone and butyric acid are known to act as agonists of one of the olfactory receptors, OR10A6. Although these substances are mainly known as odor substances and pheromone substances, the inventors have found that they have ideal effects on the skin, such as promoting the formation of the stratum corneum, inhibiting skin roughness, moisturizing, improving the skin barrier function and inhibiting the production of melanin. In particular, such an effect is exerted via the activation of OR10A6. These ingredients can also be synthesized using commercially available products.

[0070] In addition, it is believed that even if it is an ingredient other than the above, as long as it can activate OR10A6, it can also play an ideal role on the skin, such as promoting the formation of the stratum corneum, inhibiting skin roughness, moisturizing, improving the skin barrier function, and inhibiting the production of melanin. Furthermore, since OR5P2 and OR5P3 are found to be expressed in keratinocytes like OR10A6, it is expected that by activating these olfactory receptors, it can play an ideal role on the skin, such as promoting the formation of the stratum corneum, inhibiting skin roughness, moisturizing, improving the skin barrier function, and inhibiting the production of melanin. Therefore, the present invention also provides an OR10A6 activator, an OR5P2 activator, or an OR5P3 activator for promoting the formation of the stratum corneum, inhibiting skin roughness, moisturizing, improving the skin barrier function, and inhibiting the production of melanin. Examples of OR10A6 activators include agents containing substances having OR10A6 agonist effects such as 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. Among them, 3-phenylpropyl propionate, citronellol, and phenylethyl alcohol are preferred, and 3-phenylpropyl propionate can be particularly preferably used. Examples of OR5P3 activators include agents containing substances having OR5P3 agonist effects such as 1-octanol, acetophenone and coumarin as active ingredients.

[0071] OR10A6, OR5P2 or OR5P3 is a type of olfactory receptor, and is a protein encoded by the OR10A6, OR5P2 or OR5P3 gene, respectively. Olfactory receptors are present in the cilia of olfactory cells in the nasal cavity, and when the ion channels of the cells are opened by binding with odor molecules, sodium ions flow into the cells, and an action potential is generated, thereby transmitting odor information to the brain and causing the perception of odor. Although, as mentioned above, some olfactory receptors have been reported to exist in tissues other than the nose and to participate in functions other than the transmission of odor information, the inventors have discovered that OR10A6, OR5P2 or OR5P3 also exist in the skin, and further, through the activation of OR10A6, they play an ideal role on the skin, such as promoting the formation of the stratum corneum, inhibiting skin roughness, moisturizing, improving the skin barrier function, and inhibiting the production of melanin.

[0072] In one embodiment, the activation of OR10A6, OR5P2 or OR5P3 means, for example, that when the OR10A6, OR5P2 or OR5P3 activator of the present invention is added, the amount of mRNA or protein of OR10A6, OR5P2 or OR5P3 in the biological sample increases compared to the case where no addition is made. For example, the increase may be an increase with a statistically significant difference at a significance level of 5% (e.g., Student's t-test), 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, or more.

[0073] The expression of OR10A6, OR5P2 or OR5P3 can be determined by measuring the amount of mRNA or protein of OR10A6, OR5P2 or OR5P3 in a biological sample. The amount of mRNA can be determined by using methods known in the art such as quantitative PCR and Northern blotting. For example, a probe for mRNA of OR10A6, OR5P2 or OR5P3 can be used. The amount of protein can be determined by using methods known in the art such as Western blotting, immunostaining, and ICM.

[0074] In one embodiment, the activation of OR10A6, OR5P2 or OR5P3 is caused by the binding of substances (e.g., agonists of the above olfactory receptors, etc.) that have an activating effect on OR10A6, OR5P2 or OR5P3, such as 3-phenylpropyl propionate, citronellol, phenylethyl alcohol, α-cinnamyl alcohol, cyclamen aldehyde, geraniol, α-ionone, nerol, nonadecane, linalool, lyral, phenylpropanol, androstenone, androstenone, butyric acid, 1-octanol, acetophenone, coumarin, or other odor molecules, to the olfactory receptors. The activation of OR10A6, OR5P2 or OR5P3 can be induced by, for example, Na + , K + or Ca 2+ The measurement is performed by measuring the influx of cations such as Ca into cells in biological samples where these receptors are present. For example, if a candidate substance is added, the intracellular Ca 2+In the case of such an increase in cations, it can be judged that there is an activation effect of OR10A6, OR5P2 or OR5P3. For example, when a candidate substance is added, the increase in cation concentration can refer to, for example, an increase in a statistically significant difference (e.g., ANOVA analysis of variance, Scheffe method, etc.) with a significance level of 5%, or, for example, an increase of 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 or more. Although the influx of cations is not limited, it can be measured by intracellular imaging using a fluorescence microscope or the like.

[0075] When the activation effect is increased when the candidate substance is added compared to the OR10A6, OR5P2 or OR5P3 activation effect of the control, the candidate substance can also be determined as a substance having an OR10A6, OR5P2 or OR5P3 activation effect. The control OR10A6, OR5P2 or OR5P3 activation effect can be determined using an experimental system to which a control agent is added, an experimental system under the same conditions except that the candidate substance is not included, and can be determined using an experimental system including OR10A6, OR5P2 or OR5P3 knockdown cells through RNAi using shRNA, siRNA, etc., and knockout cells through genome editing using TALEN, CRISPR, etc.

[0076] The so-called biological sample is a sample containing cells whose activity of OR10A6, OR5P2 or OR5P3 can be measured, and may contain cells other than keratinocytes. As cells other than keratinocytes, there can be cited olfactory cells and other cells confirmed to have OR10A6, OR5P2 or OR5P3, or cells prepared in a manner having OR10A6, OR5P2 or OR5P3. The biological sample can be derived from any animal, but is preferably derived from humans from the viewpoint of cosmetic and pharmaceutical development. As a biological sample, it can be a cell capable of measuring the above-mentioned olfactory receptors, for example, a culture of keratinocytes or other cells whose activity of OR10A6, OR5P2 or OR5P3 can be measured, or it can be a skin sample, or a cultured skin model constructed in three dimensions can be used.

[0077] The effects of promoting stratum corneum formation, inhibiting skin roughness, moisturizing and improving skin barrier function can be confirmed, for example, by measuring cornified outer membrane (sometimes referred to as CE). For example, the amount of cornified outer membrane produced is obtained by dissolving keratinocytes with a cell lysate containing alcohols such as β-mercaptoethanol, surfactants such as SDS, etc., and counting the number of cornified outer membranes that cannot be dissolved in the solution and remain. For example, when the amount of cornified outer membrane produced measured by such operation increases the neural activation effect when the candidate substance is added compared to the control, it can be determined that the candidate substance has the effects of promoting stratum corneum formation, inhibiting skin roughness, moisturizing and improving skin barrier function. For example, the increase in the amount of cornified outer membrane produced can refer to an increase, for example, with a statistically significant difference (e.g., t-test, Dunnett's test, etc.) at a significance level of 5% compared to the control when the candidate substance is added, or an increase of 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 or more. The amount of cornified outer membrane produced in the control can be determined by an experimental system to which a control agent is added, an experimental system under the same conditions except that the candidate substance is not included, or an experimental system containing OR10A6, OR5P2 or OR5P3 knockdown cells or knockout cells. The effects of promoting stratum corneum formation, inhibiting skin roughness, moisturizing and improving skin barrier function can also be confirmed by methods other than CE determination, such as determination of proliferation markers of keratinocytes, visual inspection, TEWL, specific proteins, and lipids. For example, it can be confirmed by measuring the gene expression amount and protein production amount of TGM1 (transglutaminase-1), FLG (filagrin), HAS3 (hyaluronan synthase 3), GBA (β-glucocerebrosidase), IL-1α (interleukin 1Alpha), etc. The effect of inhibiting melanin production can be determined by measuring the mRNA amount or protein amount of genes such as IL-1α and COX2 involved in melanin production. As the determination of the amount of mRNA, quantitative PCR, Northern blotting, etc., can be used. For example, probes for mRNA of TGM1, FLG, HAS3, GBA, IL-1α, etc. can be used. Regarding the amount of protein, methods known in the art such as Western blotting, immunostaining, ICM, etc. can be used.For example, promoting stratum corneum formation, inhibiting skin roughness, moisturizing or improving skin barrier function can be confirmed by, for example, increasing the amount of mRNA or protein of TGM1, FLG, HAS3, GBA in a biological sample, and / or reducing the amount of mRNA or protein of IL-1α. For example, increase can refer to an increase in a statistically significant difference (e.g., Student's t-test, Dunnett's test) with a significance level of 5%, and / or, for example, an increase of 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 or more. For example, a reduction can refer to a reduction that is statistically significant at a significance level of 5% (e.g., Student's t-test, Dunnett's 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.

[0078] The so-called OR10A6, OR5P2 or OR5P3 knockdown cells refer to cells in which the expression of OR10A6, OR5P2 or OR5P3 is impaired or reduced by knockdown treatment of the OR10A6, OR5P2 or OR5P3 gene, for example. The knockdown treatment can be carried out using any known technique such as RNAi using shRNA or siRNA. The so-called damage or reduction of OR10A6, OR5P2 or OR5P3 expression by knockdown treatment may mean, for example, that the expression level of OR10A6, OR5P2 or OR5P3 gene and / or the amount of OR10A6, OR5P2 or OR5P3 protein in the cells subjected to knockdown treatment is reduced, for example, by a statistically significant difference (for example, t-test, etc.) at a significance level of 5%, or is reduced by more than 5%, more than 10%, more than 20%, more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90% or more than 100% as compared with non-knockdown cells.

[0079] The so-called OR10A6, OR5P2 or OR5P3 knockout cell refers to, for example, a cell in which the function of the OR10A6, OR5P2 or OR5P3 gene is completely eliminated by knockout treatment. The knockout treatment can be performed using any known technique such as genome editing using TALEN or CRISPR.

[0080] A certain embodiment of the present 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: applying one or more selected from 3-phenylpropyl propionate, citronellol, phenylethyl alcohol, α-cinnamyl alcohol, cyclamen aldehyde, geraniol, α-ionone, nerol, nonadecane, linalool, lyral, phenylpropanol, androstadienone, androsterone and butyric acid, an OR10A6 activator, an OR5P2 activator, or an OR5P3 activator (hereinafter, sometimes collectively referred to as the agent of the present invention), or a composition comprising them. The present 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 or OR5P3 of the subject. In one version of this embodiment, activation of OR10A6, OR5P2 or OR5P3 in a subject is achieved by applying 3-phenylpropyl propionate, citronellol, phenylethyl alcohol, α-cinnamyl alcohol, cyclamen aldehyde, geraniol, α-ionone, nerol, nonadecane, linalool, lyral, phenylpropanol, androstadienone, androsterone, butyric acid, 1-octanol, acetophenone, and coumarin, or an agent of the present invention, or a composition comprising the same.

[0081] Subjects to which the method of the present invention can be applied include, for example, subjects with reduced stratum corneum formation ability, subjects who have or wish to prevent or alleviate rough skin, subjects who have dry skin and need or wish to moisturize their skin, subjects with reduced skin barrier function or who wish to improve their skin barrier function, such as subjects who are concerned about sun exposure, spots, and dullness, subjects who wish to whiten their skin, subjects who need or wish to inhibit melanin production, subjects whose activity of OR10A6, OR5P2 or OR5P3 is reduced, or subjects whose olfactory receptor activity is reduced, such as subjects whose sense of smell is reduced due to aging, etc.

[0082] The method involved in this application is for beauty purposes, and medical actions performed by doctors and medical practitioners are sometimes excluded. In addition, the method involved in this application can be a method of supporting the beauty actions of the object.

[0083] A preferred embodiment is a cosmetic method for promoting stratum corneum formation, inhibiting skin roughness, moisturizing, improving skin barrier function or inhibiting melanin production in a subject, comprising: applying 3-phenylpropyl propionate. In one version of such an embodiment, promoting stratum corneum formation, inhibiting skin roughness, moisturizing, improving skin barrier function or inhibiting melanin production is achieved by activating OR10A6 in the subject.

[0084] The present invention further provides a composition for promoting stratum corneum formation, inhibiting skin roughness, moisturizing, improving skin barrier function, and / or inhibiting melanin production, comprising one or more selected from 3-phenylpropyl propionate, citronellol, phenylethyl alcohol, α-cinnamyl alcohol, cyclamen aldehyde, geraniol, α-ionone, nerol, nonadecane, linalool, lyral, phenylpropanol, androstadienone, androsterone, 1-octanol butyrate, acetophenone and coumarin, or the agent of the present invention.

[0085] The agent or composition of the present invention can be applied by any route such as transdermal, oral, transmucosal, nasal, intravenous, intraarterial, subcutaneous, 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 face, head, neck, limbs, and trunk.

[0086] The agent of the present invention can be mixed with cosmetics, medicines or quasi-drugs, respectively. These agents can be applied orally or non-orally, such as transdermally. In the case of transdermal application, it can be formulated into a skin external agent. The skin external agent is not particularly limited as long as it can be applied to the skin, and can be applied in any dosage form such as solution, emulsion, solid, semi-solid, powder, powder dispersion, water-oil two-layer separation, water-oil-powder three-layer separation, ointment, gel, aerosol, mousse, stick, etc. In the case of being formulated into a skin external agent, bases and excipients such as preservatives, emulsifiers, pH adjusters, etc. that are commonly used in skin external agents can be used. In the case of being mixed with cosmetics, it can be mixed with cosmetics for the face or body such as lotion, emulsion, beauty essence, cream, lotion, facial mask, essence, gel, foundation, isolation cream, concealer, etc., and bath agents, etc. By using a cosmetic containing the agent of the present invention, for example, the activation of OR10A6, OR5P2 or OR5P3 leads to promotion of stratum corneum formation, suppression of skin roughness, moisturizing, improvement of skin barrier function or suppression of melanin production. Therefore, the agent of the present invention can be used in cosmetics for the purpose of promoting stratum corneum formation, suppression of skin roughness, moisturizing, improvement of skin barrier function or suppression of melanin production.

[0087] The amount of the active ingredient in the agent or composition of the present invention can be arbitrarily selected from the viewpoint of promoting the formation of the stratum corneum, inhibiting skin roughness, moisturizing, improving the skin barrier function or inhibiting the production of melanin. For example, from the viewpoint of mixing as a skin external preparation, 3-phenylpropyl propionate, citronellol, phenylethyl alcohol, α-cinnamyl alcohol, cyclamen aldehyde, geraniol, α-ionone, nerol, nonadecane, linalool, lyral, phenylpropanol, androstenone, androstenone, butyric acid, 1-octanol, acetophenone, coumarin, or other active ingredients that can activate OR10A6, OR5P2 or OR5P3 can be mixed at 0.0005 to 100.0 mM. From the viewpoint of fully exerting the effect, it can be preferably mixed at 0.05 mM or more, and it can be further preferably mixed at 0.5 mM or more, for example, 1.0 mM. On the other hand, from the viewpoint of avoiding a strong odor, it is preferably blended at 10.0 mM or less, and more preferably blended at 5.0 mM or less. The above components may be combined at any ratio, and in this case, the total amount of these components is preferably within the above range.

[0088] The present invention also provides 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 the activity of OR10A6, OR5P2, or OR5P3 is used as an indicator.

[0089] In one embodiment, the screening method of the present invention comprises the following steps:

[0090] measuring the activity of OR10A6, OR5P2 or OR5P3 in a biological sample; and

[0091] When the activity of OR10A6, OR5P2 or OR5P3 is increased compared to the control, the candidate substance is determined to be a stratum corneum formation promoter, a skin roughness inhibitor, a moisturizer, a skin barrier function improver or a melanin production inhibitor.

[0092] As candidate substances, substances belonging to any library such as a cosmetic raw material library, an extract library, a drug library, a compound library, etc. Examples thereof include, but are not limited to, odor substances that react with olfactory receptors and fragrance raw materials.

[0093] Further, the present invention also provides agonists for OR10A6, OR5P2 or OR5P3 such as 3-phenylpropyl propionate, citronellol, phenylethyl alcohol, α-cinnamyl alcohol, cyclamen aldehyde, geraniol, α-ionone, nerol, nonadecane, linalool, lyral, phenylpropanol, androstenone, androstenone, butyric acid, 1-octanol, acetophenone or coumarin, etc., for promoting stratum corneum formation, inhibiting skin roughness, moisturizing, improving skin barrier function or inhibiting melanin production. Here, it is preferred that the promotion of stratum corneum formation, inhibition of skin roughness, moisturizing, improvement of skin barrier function or inhibition of melanin production is achieved by activating OR10A6, OR5P2 or OR5P3. A preferred embodiment is 3-phenylpropyl propionate for promoting stratum corneum formation, inhibiting skin roughness, moisturizing, improving skin barrier function or inhibiting melanin production, wherein the promotion of stratum corneum formation, inhibition of skin roughness, moisturizing, improvement of skin barrier function or inhibition of melanin production is achieved by activating OR10A6.

[0094] Furthermore, the present invention also provides agonists against OR10A6 such as 3-phenylpropyl propionate, citronellol, phenylethyl alcohol, α-cinnamyl alcohol, cyclamen aldehyde, geraniol, α-ionone, nerol, nonadecane, linalool, lyral, phenylpropanol, androstenone, androsterone, butyric acid, agonists against OR5P2, or 1-octanol, acetophenone, coumarin, etc., agonists against OR5P3, for use in the manufacture of an agent for promoting stratum corneum formation, inhibiting skin roughness, moisturizing, improving skin barrier function, or inhibiting melanin production.

[0095] All documents mentioned in this specification are incorporated into this specification by reference in their entirety.

[0096] The embodiments of the present invention described below are for illustrative purposes only and do not 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 present invention may be modified, such as adding, deleting and replacing the constituent elements of the present invention, without exceeding the purpose of the present invention.

[0097] Example 1

[0098] Example 1: Expression of OR10A6, OR5P2 and OR5P3 in keratinocytes

[0099] Keratinocytes (sold by Kurobo) were cultured in EPILIFE-KG2 medium (sold by Kurobo) at 37°C in a humidified atmosphere, and the culture that became confluent was collected and cDNA was prepared using Trizol (sold by NIPPON GENE) according to the instructions of VILO Master Mix (sold by Invitrogen). Real-time PCR was performed using the OR10A6 amplification primers (SEQ ID NOs. 1 and 2), OR5P2 amplification primers (SEQ ID NOs. 3 and 4), OR5P3 amplification primers (SEQ ID NOs. 5 and 6) and internal standard GAPDH amplification primers (SEQ ID NOs. 7 and 8) having the sequences shown below, and the amplification products were confirmed by electrophoresis.

[0100] OR10A6 forward: 5'-tatttacaacccaaatctg-3' (SEQ ID NO: 1)

[0101] OR10A6 reverse: 5'-tcagattgtgtgtaaaacc-3' (SEQ ID NO: 2)

[0102] OR5P2 forward: 5'-accttcatttatgtgatgc-3' (SEQ ID NO: 3)

[0103] OR5P2 reverse: 5'-aaaataacaagcatcatgag-3' (SEQ ID NO: 4)

[0104] OR5P3 forward: 5'-cagtcactctgttctatgg-3' (SEQ ID NO: 5)

[0105] OR5P3 reverse: 5'-taagctctctcttcagagc-3' (SEQ ID NO: 6)

[0106] GAPDH forward: 5'-gaaggtgaaggtcggagtc-3' (SEQ ID NO: 7)

[0107] GAPDH reverse: 5'-gaagattggtgatgggatttc-3' (SEQ ID NO: 8)

[0108] The results are shown in Figure 1 It was confirmed that OR10A6, OR5P2, and OR5P3 are all expressed in keratinocytes. Therefore, the following experiment was conducted to investigate the response in keratinocytes using OR10A6, one of these olfactory receptors, and 3PPP, its agonist.

[0109] Example 2: Observation of calcium response of cells using 3PPP

[0110] Keratinocytes (seller: クラボウ) were cultured in EPILIFW-KG2 medium (seller: クラボウ) or EPILIFW-KG2 medium containing OR10A6 siRNA (seller: Horizon) at 37°C in a humidified atmosphere for 1 day. For cells that became confluent, they were replaced with EPILIFW-KG2 containing Fura2 AM (seller: life technology) at 5 μM and cultured for 1 hour. Then, they were replaced with a buffer (pH 7.4) containing NaCl 150mM, KCl 5mM, CaCl2 1.8mM, MgCl21.2mM, HEPES25mM, and D-glucose 10mM, and 3-phenylpropyl propionate (3PPP) (seller: SigmaAldrich) was added to the cells at a concentration of 0.05 to 1mM. The intracellular Ca2+ concentration after 3PPP addition was measured by observing at wavelengths of 340 nm and 380 nm using a fluorescence microscope (seller: Nikon). 2+ The concentration change was measured as the fluorescence intensity ratio (F340 / 380) at each time point.

[0111] The results are shown in Figure 2 As shown in the figure above, if 3PPP is added, Ca 2+ The concentration increased significantly compared to the value before adding 3PPP. 2+ The increase in concentration significantly increases with the increase in the concentration of 3PPP to 0.05mM, 0.5mM, and 1.0mM, depending on the concentration. Further, the reaction using 3PPP is confirmed to be significantly lower in OR10A6 knockdown keratinocytes by the following figure. Through the above experiments, it can be seen that OR10A6 present in keratinocytes is activated by 3PPP as its agonist. Therefore, the following experiment was conducted to investigate the effects on the skin caused by activating OR10A6 by 3PPP.

[0112] Example 3: Effects of OR10A6 activation on promoting stratum corneum formation, inhibiting skin roughness, moisturizing and improving skin barrier function 1

[0113] Keratinocytes (seller: クラボウ) were cultured in EPILIFW-KG2 medium (seller: クラボウ) or EPILIFW-KG2 medium containing OR10A6 siRNA (seller: Horizon) at 37°C in a humidified atmosphere for 1 day. For cells that became confluent, 3-phenylpropyl propionate (3PPP) (seller: SigmaAldrich) was added to the cells at a concentration of 1 mM and further cultured for 3 days. After 3 days, the cultured cells were recovered by treatment with trypsin (seller: Nakalä Tesque Co., Ltd.), and the total number of cells was counted under an optical microscope, and the number of dead cells stained with trypan blue (seller: Nakalä Tesque Co., Ltd.) was further counted, thereby obtaining the number of live cells as the total number of cells minus the number of dead cells. Then, the cells were centrifuged and replaced with a buffer solution containing 1% β-mercaptoethanol, 1% SDS (sodium lauryl sulfate), and 20 mM tri-HCl (pH 7.5), and the cells were lysed at 95°C for 15 minutes, and the number of keratinocytes with cornified outer membranes that could not be dissolved in the solution and remained was counted using an optical microscope. As an index of the effects of promoting stratum corneum formation, inhibiting skin roughness, moisturizing, and improving skin barrier function, the amount of cornified outer membrane produced was calculated as the number of keratinocytes with cornified outer membranes / the number of viable cells × 100, and this value was expressed as the CE production rate (%).

[0114] The results are shown in Figure 3 In the figure, the CE production rate in keratinocytes was significantly increased by 3PPP (left figure). However, for OR10A6 knockout keratinocytes, even with the addition of 3PPP, no significant increase was observed in the production of cornified outer membranes. Further, without the addition of 3PPP, the CE production rate in OR10A6 knockout keratinocytes was significantly reduced compared with OR10A6 non-knockout keratinocytes (right figure). Therefore, it is suggested that CE production is promoted by the activation of OR10A6.

[0115] The above results indicate that OR10A6, OR5P2, and OR5P3 exist in keratinocytes and that activation of OR10A6 by 3PPP promotes CE production.

[0116] Example 4: Effects of OR10A6 activation on promoting stratum corneum formation, inhibiting skin roughness, moisturizing and improving skin barrier function 2

[0117] Example 3 showed the possibility that 3PPP promotes the formation of CE via activation of olfactory receptors such as OR10A6. In this example, the expression of the TGM1 gene encoding transglutaminase 1, an enzyme that promotes the formation and maturation of CE via activation of OR10A6, was measured.

[0118] 4-1: Cell culture

[0119] HaCaT cells were plated in 24-well plates (Cat No. 3526, Corning, USA) at a density of 10.0 × 10 4 The density of cells / well was inoculated in 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 cultured for 24 hours in a CO2 incubator (CO2 concentration 5%, 37°C). After the culture medium was removed, it was replaced with a culture medium to which 3PPP was added in a final concentration of 0.05mM, and further cultured in a CO2 incubator for 48 hours. A culture medium not containing 3PPP was used as a control.

[0120] 4-2: RNA extraction / purification, quantification and purity determination

[0121] RNA was extracted / purified using PureLinkTM RNA Mini Kit (Cat No. 12183018A, Invitrogen, USA), and a portion of the purified RNA was taken to a UV-transmissive 96-well plate, diluted 10-fold with Tris-EDTA Buffer, and the absorbance (OD230, OD260, OD280) at 230 nm, 260 nm, and 280 nm was measured using a microplate reader (SPARK (registered trademark) 10M TECAN, Switzerland). RNA concentration was calculated using OD260, and RNA concentration was adjusted to 10 μg / mL by dilution with TE buffer.

[0122] 4-3: Gene expression analysis using real-time PCR

[0123] RNA was reverse transcribed using SuperScript (trademark) IV VILOTM Master Mix with ezDNase (Cat No. 11766050, Invitrogen, USA). 4 μL of SuperScript (trademark) IV VILOTM Master Mix and 6 μL of nuclease-free water were added to each well of an 8-tube tube, and cDNA was synthesized by heating at 25°C for 10 minutes, at 50°C for 10 minutes, and at 85°C for 5 minutes using real-time PCR (QuantStudio (registered trademark) 3, Applied Biosystems, USA). 10 μL of TaqMan (registered trademark) Fast Advanced Master Mix (Cat No. 4444557, Applied Biosystems, USA), 1 μL of TaqMan Gene Express, 7 μL of UltraPureTM Distilled Water (Invitrogen, Cat No. 10977-015, USA) and 2 μL of cDNA were added to each well of the PCR plate and sealed. Real-time qPCR was performed using primers for TGM1 (Hs00165929_m1) and primers for GAPDH as an internal standard gene (Hs02786624_g1), and after calculating the Threshold Cycle (Ct) value of TGM1 in the 3PPP-added medium, the Ct value was corrected by GAPDH and set to a ΔCt value, and, assuming that the gene amount was doubled per cycle, the gene expression amount in the 3PPP-added medium was calculated when the gene expression amount of the control was set to 1. In the analysis of gene expression levels, the average value of 3 wells in a 24-well plate was used for one treatment group.

[0124] The results are shown in Table 1 below.

[0125] Table 1: Promotion of TGM1 expression caused by 3PPP addition

[0126]

[0127] (Compared with the non-additive group in the paired sample T test (two-sided), the ratio is recorded with the non-additive group set to 1.)

[0128] As shown in Table 1, the addition of 3PPP significantly increased the gene expression of TGM1. 3PPP activated OR10A6 and promoted the production of TGM1, thereby promoting the formation and maturation of CE, suggesting the possibility of promoting stratum corneum formation, inhibiting skin roughness, moisturizing and improving skin barrier function.

[0129] Example 5: Effects of OR10A6 activation on promoting stratum corneum formation, inhibiting skin roughness, moisturizing and improving skin barrier function

[0130] In order to confirm the effects of OR10A6 activation on the formation of stratum corneum, the inhibition of skin roughness, moisturizing, and the improvement of skin barrier, the following experiment was conducted using other genes involved in these functions. More specifically, as NMF, the expression levels of FLG (Filaggrin) encoding the fibroin protein involved in the moisture retention function of the skin, HAS3 (Hyaluronan Synthase 3) encoding the synthase of hyaluronic acid produced in epidermal cells and involved in the moisture retention of the skin, and GBA (β-enzyme glucocerebrosidase) encoding the enzyme producing ceramide EOP (ceramide 1) related to the barrier function of the skin were measured.

[0131] Instead of using the primer for TGM1 (Hs00165929_m1), primers for FLG (Hs00856927_g1), human hyaluronan synthase 3 (HAS3, Assay ID. Hs00193436_m1), and human β-glucocerebrosidase (GBA, Assay ID. Hs00986836_g1) were used. In addition, the expression levels of these genes in the culture medium containing or not containing 0.05 mM 3PPP were measured using the same materials and methods as in Example 4.

[0132] The results are shown in Tables 2 to 4 below.

[0133] Table 2: Promotion of FLG expression caused by 3PPP addition

[0134]

[0135] (Compared with the non-additive group in the paired sample T test (two-sided), the ratio is recorded with the non-additive group set to 1.)

[0136] Table 3: Promotion of HAS3 expression caused by 3PPP addition

[0137]

[0138] (Compared with the non-additive group in the paired sample T test (two-sided), the ratio is recorded with the non-additive group set to 1.)

[0139] Table 4: Promotion of GBA expression caused by 3PPP addition

[0140]

[0141] (Compared with the non-additive group in the paired sample T test (two-sided), the ratio is recorded with the non-additive group set to 1.)

[0142] As shown in Tables 2 to 4, the addition of 3PPP significantly increased the gene expression of FLG, HAS3, and GBA. Based on the above results, the activation of OR10A6 by 3PPP and the like promotes the expression of these genes in addition to TGM1, suggesting the possibility of promoting stratum corneum formation, inhibiting skin roughness, moisturizing, and improving skin barrier function.

[0143] Example 6: Effects of OR10A6 activation on promoting stratum corneum formation, inhibiting skin roughness, moisturizing and improving skin barrier function

[0144] In order to investigate whether the same effect is observed with respect to OR10A6 activators other than 3PPP, citronellol and phenylethanol (Non-Patent Document 5), which are known substances that react with OR10A6, were used to investigate whether these substances have a stratum corneum-forming effect similar to 3PPP. More specifically, the TGM1 gene expression level was measured using the same materials and methods as in Example 4, except that citronellol (CAS No.: 106-22-9, Wako 037-05993) or phenylethanol (CAS No.: 60-12-8, TCI P0084) was used at the following concentrations instead of 3PPP.

[0145] The results are shown in Tables 5 and 6 below.

[0146] Table 5: Promotion of TGM1 expression caused by citronellol addition

[0147]

[0148] (Compared with the non-additive group in the paired sample T test (two-sided), the ratio is recorded with the non-additive group set to 1.)

[0149] Table 6: Promotion of TGM1 expression caused by the addition of phenylethanol

[0150]

[0151] (Compared with the non-additive group in the paired sample T test (two-sided), the ratio is recorded with the non-additive group set to 1.)

[0152] According to the results in Table 6, the expression of TGM1 was significantly increased not only by 3PPP but also by citronellol and phenylethanol, other OR10A6 activators.

[0153] Example 7: Effects of OR10A6 activation on promoting stratum corneum formation, inhibiting skin roughness, moisturizing and improving skin barrier function

[0154] For citronellol and phenylethanol, which showed effects in Example 6, the following experiment was conducted using GBA in the same manner as in Example 5. More specifically, the GBA gene expression level was measured using the same materials and methods as in Example 5, except that citronellol (CAS No.: 106-22-9, Wako 037-05993) or phenylethanol (CAS No.: 60-12-8, TCI P0084) was used at the following concentrations instead of 3PPP.

[0155] The results are shown in Tables 7 and 8 below.

[0156] Table 7: Promotion of GBA expression caused by citronellol addition

[0157]

[0158] (Compared with the non-additive group in the paired sample T test (two-sided), the ratio is recorded with the non-additive group set to 1.)

[0159] Table 8: Promotion of GBA expression caused by the addition of phenylethanol

[0160]

[0161] (Compared with the non-additive group in the paired sample T test (two-sided), the ratio is recorded with the non-additive group set to 1.)

[0162] According to the results in Table 7, similarly to Example 6, even with citronellol and phenylethanol, which are OR10A6 activators other than 3PPP, the expression of GBA increased significantly or tended to increase significantly.

[0163] Example 8: Effects of OR10A6 activation on promoting stratum corneum formation, inhibiting skin roughness, moisturizing and improving skin barrier function

[0164] For citronellol, which showed effects in Examples 6 and 7, the following experiment was conducted using HAS3 in the same manner as in Example 5. More specifically, the HAS3 gene expression level was measured using the same materials and methods as in Example 5, except that citronellol (CAS No.: 106-22-9, Wako 037-05993) was used at the following concentrations instead of 3PPP.

[0165] The results are shown in Table 9 below.

[0166] Table 9: Promotion of HAS3 expression caused by citronellol addition

[0167]

[0168] (Compared with the non-additive group in the paired sample T test (two-sided), the ratio is recorded with the non-additive group set to 1.)

[0169] Embodiment 9:

[0170] According to the above suggestions, OR10A6 is activated to promote stratum corneum formation, inhibit skin roughness, moisturize and improve skin barrier function. However, in order to confirm other effects, the expression level of IL-1α (Interleukin 1 Alpha) encoding IL-α, which is known to be an inflammatory cytokine and promotes melanin production, was measured. More specifically, instead of the primer of TGM1 (Hs00165929_m1), the primer of IL-1α (Hs00174092_m1) was used. In addition, the gene expression level of IL-1α in the culture medium containing or not containing the following concentration of 3PPP was measured using the same materials and methods as Example 4. It is known that IL-1α (Interleukin 1 Alpha) is a gene encoding IL-1α protein, which is an inflammatory cytokine. IL-1α causes excessive production of melanin by increasing tyrosinase activity.

[0171] The results are shown in Table 10 below.

[0172] Table 10: Inhibition of IL-1α expression caused by 3PPP addition

[0173]

[0174] (Dunnett's test, recorded as the ratio of the non-added group set to 1)

[0175] As shown in Table 10, the addition of 3PPP reduced the gene expression of IL-α in a concentration-dependent manner. This result suggests that the activation of OR10A6 not only contributes to the promotion of stratum corneum formation, suppression of skin roughness, moisturizing, and improvement of skin barrier function by promoting the expression of TGM1, FLG, HAS3, and GBA, but also contributes to the suppression of skin roughness and the possibility of suppressing melanin production by suppressing the expression of IL-α.

[0176] From the above results, it can be expected that the activation of OR10A6, OR5P2 and OR5P3 will bring about desirable effects on the skin, such as promoting stratum corneum formation, inhibiting skin roughness, moisturizing, improving skin barrier function, and inhibiting melanin production, and these effects are achieved by ingredients that activate OR10A6, OR5P2, and OR5P3. For example, as ingredients that activate OR10A6, 3-phenylpropyl propionate, citronellol, phenylethyl alcohol, α-cinnamyl alcohol, cyclamen aldehyde, geraniol, α-ionone, nerol, nonadecane, linalool, lyral, phenylpropanol, androstenone, androsterone, and butyric acid are known, and as ingredients that activate OR5P3, 1-octanol, acetophenone, and coumarin are known, so the effects brought about by these ingredients 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 was used as an indicator.

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