CD39 expression promoter and composition for enhancing sedative effect of skin immune cell

Abstract

To provide a CD39 expression promoter or the like that can exhibit a superior CD39 expression promoting action.SOLUTION: A CD39 expression promoter comprises at least one selected from compounds with a flavanone skeleton, compounds with in vivo vitamin A activity, compounds with a chalconoid skeleton, lignans, and stilbenoids.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a CD39 expression promoter and a composition that enhances the calming effect of skin immune cells. [Background technology]

[0002] The epidermis, the outermost layer of the skin, is a thin film about 0.2 mm thick that acts as a barrier to prevent the intrusion of foreign substances from the outside and the evaporation of water. The epidermis is made up of four layers from the outside: the stratum corneum, stratum granulosum, stratum spinosum, and stratum basale, and is composed of cells such as keratinocytes, granular cells, Langerhans cells (which are dendritic cells within the epidermis), and melanocytes.

[0003] Recent research has revealed that Langerhans cells play an important role in skin immune function. The surface of Langerhans cells is covered with an enzyme called CD39, which acts as an ATPase. It is believed that CD39 reduces the amount of ATP, which is a stimulus response factor, and contributes to the calming of inflammation and reduces skin damage caused by ultraviolet rays and physical stimuli. It is known that the self-defense function of Langerhans cells, which calms stimulus response factors, is reduced by aging, stress, etc. In skin with a reduced self-defense function due to aging, stress, etc., skin problems such as rough skin and reduced elasticity occur due to a decrease in the barrier function of the stratum corneum and destruction of collagen fibers in the dermis.

[0004] Ingredients that increase the expression level of CD39 and ingredients that inhibit the decrease or activate Langerhans cells have been studied. Patent Document 1 discloses a CD39 gene expression promoter that contains three ingredients: carboxymethyl β-glucan or a salt thereof, polyoxyethylene / polyoxypropylene random copolymer dimethyl ether, and rose water. Patent Document 2 discloses a CD39 expression promoter in skin Langerhans cells that contains an extract of Aster staghorn. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Special Publication No. 2017-511790 [Patent Document 2] Patent No. 6887649 Summary of the Invention [Problem to be solved by the invention]

[0006] The present invention provides a CD39 expression promoter capable of exerting an excellent CD39 expression promoting effect and a composition for enhancing the calming effect of skin immune cells. [Means for solving the problem]

[0007] As a result of studies conducted by the present inventors to achieve the above object, they have newly discovered that specific compounds, such as compounds having a flavanone skeleton, exhibit excellent CD39 expression promoting activity. Various aspects for solving the above problems will be described below.

[0008] The CD39 expression promoter of aspect 1 is characterized by containing at least one selected from a compound having a flavanone skeleton, a compound having vitamin A activity in the body, a compound having a chalconoid skeleton, lignans, and stilbenoids.

[0009] Aspect 2 relates to the CD39 expression promoting agent according to Aspect 1, which promotes the expression of CD39 in skin Langerhans cells. In a third aspect, in the CD39 expression promoter according to the first or second aspect, the compound having a flavanone skeleton is at least one selected from flavanones and glycosides of flavanones.

[0010] Aspect 4 is the CD39 expression promoter according to any one of Aspects 1 to 3, wherein the compound having vitamin A activity in the body is at least one selected from retinal, retinol, retinoic acid, and provitamin A.

[0011] Aspect 5 is the CD39 expression promoter according to any one of Aspects 1 to 4, wherein the stilbenoid is at least one selected from resveratrol, piceatannol, pterostilbene, and oligomers thereof.

[0012] The composition of aspect 6 for enhancing the calming effect of skin immune cells is characterized in that it contains at least one selected from a compound having a flavanone skeleton, a compound having vitamin A activity in the body, a compound having a chalconoid skeleton, lignans, and stilbenoids. Effect of the Invention

[0013] The present invention can exert an excellent effect of promoting CD39 expression or enhancing the calming effect of skin immune cells. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] An embodiment of the CD39 expression promoter according to the present invention will be described. The CD39 expression promoter of this embodiment contains at least one selected from a compound having a flavanone skeleton, a compound having vitamin A activity in the body, a compound having a chalconoid skeleton, lignans, and stilbenoids. These compounds may be used alone or in appropriate combination of two or more.

[0015] (Active ingredient) Examples of compounds having a flavanone skeleton include flavanones and glycosides of flavanones in which the 7-position is glycosylated with a disaccharide.

[0016] Specific examples of compounds having a flavanone skeleton include flavanones such as 7-hydroxyflavanone, butyne, eriodictyol, hesperetin, isosakuranetin, homoeriodictyol, naringenin, sakuranetin, sterubin, and pinocembrin, and 3-hydroxyflavanone (flavanonol).

[0017] Among these, flavanones and glycosides of flavanones are preferred from the viewpoint of having a more excellent CD39 expression promoting effect, and naringenin, 7-hydroxyflavanone, isosakuranetin, and hesperetin are more preferred.

[0018] Specific examples of compounds that have vitamin A activity in the body include retinoids such as retinol (vitamin A1), retinal, retinoic acid, tretinoin, and vitamin A derivatives, and provitamin A such as α-carotene, β-carotene, γ-carotene, β-cryptoxanthin, and retinyl esters.

[0019] Among these, retinal, retinol, retinoic acid, and provitamin A are preferred from the viewpoint of having a more excellent CD39 expression promoting effect, and retinal, retinol, retinoic acid, and β-carotene are more preferred.

[0020] Examples of compounds having a chalconoid skeleton include chalcone and its derivatives, i.e., chalcones. Specific examples of compounds having a chalconoid skeleton include chalcone, isoliquiritigenin, butein, carthamin, rottlerin, and 2,2',4'-trihydroxychalcone.

[0021] Among these, isoliquiritigenin, butein, and 2,2',4'-trihydroxychalcone are preferred from the viewpoint of having a more excellent CD39 expression promoting effect. Lignans refer to a group of natural organic compounds in which a benzene ring and three carbon chains, i.e., phenylpropanoid C6-C3 units, are bonded together in two or three units. Specific examples of lignans include sesamin, sesamol, sesaminol, sesamolinol, sesamolin, honokiol, magnolol, pinoresinol, podophyllotoxin, steganacin, enterodiol, enterolactone, lariciresinol, secoisolariciresinol, matairesinol, hydroxymatairesinol, syringaresinol, and arctigenin. Among these, sesamin, sesamol, honokiol, and magnolol are preferred from the viewpoint of having a more excellent CD39 expression promoting effect.

[0022] Specific examples of stilbenoids include resveratrol, piceatannol, pterostilbene, pinosylvin, etc. Stilbenoids may be in the form of aglycones, oligomers thereof, glycosides thereof, etc.

[0023] Among these, resveratrol, piceatannol, pterostilbene, their oligomers, their glycosides, etc. are preferred from the viewpoint of having a more excellent CD39 expression promoting effect.

[0024] The formulation of at least one selected from the group consisting of compounds having a flavanone skeleton, compounds having vitamin A activity in the body, compounds having a chalconoid skeleton, lignans, and stilbenoids is not particularly limited, and may be, for example, an extract from a natural material, a refined product, a biochemical product, or a chemically synthesized product. In addition, a commercially available product may also be used.

[0025] (Other active ingredients) Other active ingredients having the CD39 expression promoting effect according to the present invention include the following: daidzein, an isoflavone, farnesol, a sesquiterpene, ginsenoside Rg1, a triterpene, kojibiose, an oligosaccharide, and urolithin A, a tannin.

[0026] (action) The CD39 expression promoter of this embodiment has an excellent effect of promoting the expression level of the CD39 gene by using each of the above-mentioned compounds. In particular, it promotes the expression level of the CD39 gene in skin Langerhans cells. The surface of Langerhans cells is covered with an enzyme called CD39 that acts as an ATPase. In the CD39 expression promoter of this embodiment, this CD39 acts as a calming enzyme to reduce the amount of ATP, which is an extracellular stimulus response factor. It is expected that the CD39 expression promoting effect will contribute to improving the calming effect of skin immune cells and calming inflammatory stimulus responses through the decomposition of extracellular ATP by Langerhans cells. As a result, it is expected to exert an effect of reducing skin disorders caused by, for example, ultraviolet rays and physical stimuli. In addition, it is known that in skin in which the self-defense function of calming inflammatory stimulus responses by Langerhans cells is reduced due to, for example, aging, stress, etc., the barrier function of the stratum corneum is reduced, and collagen fibers in the dermis are destroyed, resulting in skin disorders such as rough skin and reduced elasticity. Therefore, CD39 expression promoters are expected to have the effect of suppressing, reducing, and preventing the worsening of various symptoms such as inflammation, rough skin, loss of skin elasticity, skin aging, spots, wrinkles, sagging, and thinning.

[0027] (Usage form) Specific formulation forms of the CD39 expression promoter of the present embodiment can be applied as skin external preparations, medicines, quasi-drugs, cosmetics, food and beverages, and test and research reagents, etc., for the purpose of obtaining the above-mentioned action and effect. Skin is mainly composed of the epidermis, basement membrane, and dermis from the outside. The CD39 expression promoter of the present embodiment can effectively exert its action and effect, particularly by applying it to the skin surface. Therefore, among the above-mentioned formulation forms, it is preferable to apply it as a cosmetic or skin external preparation for the purpose of obtaining the CD39 expression promoting action. In addition, when the CD39 expression promoter of the present embodiment is applied as a skin external preparation, cosmetics, and food and beverages, in order to distinguish it from conventional products, it is preferable to attach a label indicating that the purpose is to obtain the above-mentioned action and effect, such as promotion of CD39 expression, improvement of the calming action of skin immune cells, and calming of inflammatory stimulus response, as well as the effect of suppressing, reducing, and preventing the worsening of each symptom, such as inflammation, rough skin, loss of skin elasticity, skin aging, spots, wrinkles, sagging, and thinning, associated with these action and effect.

[0028] When the CD39 expression promoter of the present embodiment is applied to a cosmetic, it can be produced by blending it with a cosmetic base material. The cosmetic may be in any form such as a milky liquid, cream, or powder. By applying such a cosmetic to the skin, the effect of promoting CD39 expression can be obtained. The cosmetic base material is generally blended in common with cosmetics, and for example, oil, purified water, and alcohol are the main components, and at least one selected from surfactants, moisturizers, antioxidants, thickeners, antiseborrheic agents, blood circulation promoters, whitening agents, pH adjusters, color pigments, preservatives, and fragrances are appropriately blended.

[0029] When the CD39 expression promoter of this embodiment is used as a drug or quasi-drug, it is preferably in the form of a skin external preparation. The dosage form is not particularly limited, but examples thereof include ointments, liquids, sprays, sheets, powders, and powders. In addition, excipients, bases, emulsifiers, solvents, stabilizers, and the like may be added as additives. In addition, when the CD39 expression promoter of this embodiment is used as a drug or quasi-drug, in addition to administration by oral ingestion (oral ingestion), any administration method such as subcutaneous injection, intravascular administration, and transdermal administration can be adopted. The dosage form is not particularly limited, but examples thereof include powders, powders, granules, tablets, capsules, pills, suppositories, liquids, and injections. In addition, excipients, bases, emulsifiers, solvents, stabilizers, and the like may be added as additives.

[0030] When the CD39 expression promoter of this embodiment is applied to food and beverages, the CD39 expression promoter can be used as the food and beverage itself, or can be blended with various food or beverage ingredients. The form of the food and beverage is not particularly limited, and may be any of liquid, powder, gel, and solid, and the dosage form may be any of tablets, capsules, granules, and drinks. Among them, capsules are preferable because they have low hygroscopicity. The food and beverage may be appropriately blended with other ingredients such as gelling agent-containing foods, sugars, flavors, sweeteners, oils and fats, base materials, excipients, food additives, secondary materials, and bulking agents.

[0031] The uses of the food composition are not particularly limited, and include so-called (1) general foods, (2) health foods such as nutritional supplements, health supplements, nutritionally adjusted foods, and supplements, (3) health functional foods such as foods for specified health uses, nutritional functional foods, functional foods, and functional foods, and (4) special purpose foods such as foods for patients. When the uses of foods and beverages are indicated, indications stipulated by various laws, enforcement regulations, guidelines, etc. are included. When the uses of foods and beverages are indicated, indications of the uses of foods and beverages include indications on packages such as wrapping and containers, as well as indications on advertising media such as pamphlets. When the uses of CD39 expression promoters are indicated, indications of CD39 expression promoting action, improvement of the calming action of skin immune cells, calming of inflammatory stimulus responses, etc. are included. In addition, indications of the suppression, reduction, and prevention of worsening of each symptom such as inflammation, rough skin, loss of skin elasticity, skin aging, spots, wrinkles, sagging, and thinning associated with these actions and effects are included.

[0032] The CD39 expression promoter of this embodiment may be used as an experimental or research reagent in the form of a reagent having a CD39 expression promoting effect, and is preferably used in the fields of elucidation of the mechanism of physiological actions related to CD39 expression, research and development of treatments for various symptoms, and the like.

[0033] The content of the above-mentioned active ingredient (solid content) in the CD39 expression promoter of this embodiment is appropriately determined depending on the other ingredients used, such as the solvent, the purpose, etc., but from the viewpoint of enhancing the CD39 expression promoting effect of the active ingredient and from the viewpoint of economy, it is preferably contained at 0.0001 to 10 mass%.

[0034] The effect of the CD39 expression promoter of this embodiment will be described. (1) The CD39 expression promoter of this embodiment is configured to contain at least one selected from the above-mentioned compounds having a flavanone skeleton, compounds having vitamin A activity in the body, compounds having a chalconoid skeleton, lignans, and stilbenoids. Therefore, it exhibits an excellent CD39 expression promoting effect. In particular, it promotes the expression of CD39 in skin Langerhans cells. By promoting CD39 expression, Langerhans cells contribute to improving the calming effect of skin immune cells and calming the inflammatory stimulus response through the decomposition of extracellular ATP. It is expected to exert an effect of reducing skin disorders caused by ultraviolet rays and physical stimuli. In addition, the CD39 expression promoter is expected to exert an effect of suppressing, reducing, and preventing the worsening of each symptom such as inflammation, rough skin, loss of skin elasticity, skin aging, spots, wrinkles, sagging, and thinning.

[0035] The above embodiment may be modified as follows: The above embodiment and the following modifications may be combined with each other to the extent that no technical contradiction occurs. In the above embodiment, a compound having a flavanone skeleton was used as an active ingredient in order to obtain an effect of promoting CD39 expression.

[0036] In addition, the composition may be configured as a composition that enhances the calming effect of skin immune cells, a composition that enhances the calming effect of inflammatory stimulus responses, a skin aging inhibitor, or a Langerhans cell activator, which contains at least one active ingredient selected from a compound having a flavanone skeleton, a compound having vitamin A activity in the body, a compound having a chalconoid skeleton, lignans, and stilbenoids. EXAMPLES

[0037] The configuration and effects of the present invention will be specifically described below using examples, but the present invention is not limited to these examples. <Test Example 1: Measurement of CD39 gene expression level> Using the method and test samples described below, the increased level of CD39 gene expression due to the addition of each test sample was measured.

[0038] (cell) THP-1 cells (hereafter referred to as "THP-1") (JCRB Cell Bank), which are human monocyte-macrophage-like cells used as an alternative to Langerhans cells, were passaged in RPMI 1640 medium (containing 10% FBS and 1% penicillin / streptomycin) and used in the study.

[0039] (Test Sample) The compounds shown in Tables 1 and 2 below were used. Compounds that have the effect of increasing the expression level of the CD39 gene include flavanones, chalcones, isoflavones, sesquiterpenes (C15), triterpenes (C30), compounds with vitamin A activity in the body (carotenoids, retinoids), lignans, stilbenoids, oligosaccharides, and tannins. More specifically, naringenin, 7-hydroxyflavanone, isosakuranetin, hesperetin, isoliquiritigenin, butein, 2,2',4'-trihydroxychalcone, β-carotene, retinal, retinol, retinoic acid, sesamol, honokiol, sesamin, magnolol, resveratrol, pterostilbene, piceatannol, daidzein, farnesol, ginsenoside Rg1, kojibiose, and urolithin A were used. Daidzein, farnesol, ginsenoside Rg1, kojibiose, and urolithin A are shown as reference examples.

[0040] Compounds that do not increase the amount of CD39 gene expression include flavan-3-ols, flavones, flavonols, dihydroflavonols, dihydrochalcones, anthocyanins, monoterpenes (C10), diterpenes (C20), steroids, monolignans, coumarins, shikimic acid pathway-related compounds, polyketides, carboxylic acids, indole alkaloids, isoquinoline alkaloids, pyridine alkaloids, purine alkaloids, capsaicins, monosaccharides, amino acid fatty acids, inositols, isothiocyanates, polyamines, guanidines, cholines, coenzymes, and galloyl derivatives. In addition, isoflavones other than those mentioned above, sesquiterpenes (C15), triterpenes (C30), carotenoids, oligosaccharides, and tannins are also included. More specifically, epicatechin, epicatechin gallate, epigallocatechin, luteolin, baicalin, kaempferol, fisetin, quercetin, isorhamnetin, silibinin, taxifolin, dihydrokaempferol, phloretin, phloridzin, cyanidin 3-glucoside, cyanine chloride, ideain chloride, biochanin A, formononetin, geraniol, D-limonene, cineole, caryophyllene, abietic acid, carnosic acid, limonin, oleanolic acid, astaxanthin, timosaponin A-III, caffeic acid phenethyl ester, rosmarinic acid, isochlorogenic acid A, p-coumaric acid, umbelliferone, coumarin, curcumin, The following substances were used: amine, [6]-gingerol, emodin, α-mangostin, DL-malic acid, L-lactic acid, citric acid, sodium butyrate, melatonin, indole-3-carbinol, brassinin, berberine, nicotinic acid, nicotinamide, caffeine, capsaicin, D(-)-mannitol, D-glucosamine, xylitol, D(+)-trehalose, palatinose, 5-aminolevulinic acid, phytic acid, D-pinitol, DL-sulforaphane, allyl isothiocyanate, spermidine, spermine, creatine, guanidinium chloride, L-arginine, α-glycerophosphocholine, pyrroloquinoline quinone disodium, and bergenin.In addition, for compounds that do not show the effect of increasing the expression level of the CD39 gene, data attachment is omitted for compounds other than flavonols, dihydroflavonols, and capsaicins.

[0041] [Table 1]

[0042] [Table 2] (Cell Test) 2 x 10 THP-1 cells in a 96-well plate 5 The cells were seeded at 100 cells / well, a test sample was added, and the cells were cultured for 24 hours in a CO2 incubator (37°C, 5% CO2).

[0043] After culturing, the cells were harvested and TM RNA lysates from the cells were prepared using the Direct RNA Prep Kit for RT-PCR (Real Time) (Takara Bio). RT-qPCR was performed using the One Step TB Green Prime Script PLUS RT-PCR Kit (Perfect Real Time) (Takara Bio). The following primers were used for PCR.

[0044] h-ENTPD1(CD39)-forward: SEQ ID NO: 1 h-ENTPD1(CD39)-reverse: SEQ ID NO: 2 h-RPL37A-forward: SEQ ID NO:3 h-RPL37A-reverse: SEQ ID NO: 4 h-RPS9-forward: SEQ ID NO:5 h-RPS9-reverse: SEQ ID NO:6 The activity of the test samples shown in Table 3 was confirmed at n=1, and for samples that exceeded 2SD from the control, the presence or absence of an increase in CD39 gene expression was evaluated at n=3, at a final concentration of 30 μM (6 μM for compounds that could not be evaluated at 30 μM). The gene expression level was corrected with h-RPL37A or h-RPS9 gene, and a relative value was calculated with the control average set at 1. The average value and standard deviation were calculated from the obtained values. The results are shown in Table 3.

[0045] Statistical analysis was performed by F-test between the control and test sample treated groups, and if the variances were equal, a Student's t-test was performed, and if the variances were unequal, a Welch's t-test was performed. A significance level of less than 5% (p<0.05) in a two-sided test was determined to indicate an enhancement effect on CD39 expression. Statistical analysis was performed using Excel Statistics 2016 (SSRI). Regarding the statistical analysis results, * indicates P value <0.05, ** indicates P value <0.01.

[0046] [Table 3] As shown in Table 3, the test samples belonging to flavanones, chalcones, compounds with vitamin A activity in the body, lignans, and stilbenoids all showed a significant increase compared to the control. Daidzein, farnesol, ginsenoside Rg1, kojibiose, and urolithin A also showed a significant increase compared to the control. Fisetin, silibinin, taxifolin, dihydrokaempferol, and capsaicin did not show a significant increase compared to the control. In other words, it was confirmed that compounds with a flavanone skeleton, compounds with vitamin A activity in the body, compounds with a chalconoid skeleton, lignans, or compounds with a specific common structure belonging to stilbenoids have an effect of increasing CD39 expression.

Claims

1. A CD39 expression promoter comprising at least one member selected from the group consisting of compounds having a flavanone skeleton, compounds having vitamin A activity in the body, compounds having a chalconoid skeleton, lignans, and stilbenoids.

2. The CD39 expression promoter according to claim 1, which promotes the expression of CD39 in skin Langerhans cells.

3. The CD39 expression promoter according to claim 1, wherein the compound having a flavanone skeleton is at least one selected from flavanones and glycosides of flavanones.

4. The CD39 expression promoter according to claim 1 , wherein the compound having vitamin A activity in the body is at least one selected from the group consisting of retinal, retinol, retinoic acid, and provitamin A.

5. The CD39 expression promoter according to claim 1, wherein the stilbenoid is at least one selected from the group consisting of resveratrol, piceatannol, pterostilbene, and oligomers thereof.

6. A composition that enhances the calming effect of skin immune cells, characterized by containing at least one selected from a compound having a flavanone skeleton, a compound having vitamin A activity in the body, a compound having a chalconoid skeleton, lignans, and stilbenoids.

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