Expression enhancer of epidermal moisturizing factor or barrier function factor

Abstract

To provide an enhancer for the expression of epidermal moisturizing factors or barrier function factors.SOLUTION: An enhancer for the expression of epidermal moisturizing factors or barrier function factors contains an extract from the bark of Albizia julibrissin.SELECTED DRAWING: None

Description

[Technical Field]

[0001] A technique is disclosed relating to an expression enhancer for an epidermal moisturizing factor or barrier function factor. [Background technology]

[0002] Silk trees are deciduous tall trees of the legume family that grow naturally in sunny wetlands such as forest edges and fields in mountainous areas south of the Tohoku region in Japan, but are also distributed and cultivated overseas in China, Southeast Asia, and other regions. The bark is harvested during the flowering season, washed with water, sun-dried, and chopped into appropriate lengths. This bark is called "silk bark" and is commonly taken as a diuretic, edema-relieving, and tonic. It is also used externally as a cold compress for swelling, bruises, and joint pain, or as a bath additive. Silk bark is known to contain triterpene saponins (numerous julibrosides), flavonoids (gerardone, isoocanin, luteolin, etc.), tannins, and other compounds (Non-Patent Document 1).

[0003] It is known that an extract of Albizia julibrissin can be used, for example, on skin cells that exhibit irregular or abnormal clock gene expression rhythms compared to normal clock gene expression rhythms (Patent Document 1). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] International Publication No. 2021 / 005941 [Non-patent literature]

[0005] [Non-Patent Document 1] Wakanyaku No.759 (2016.8) Summary of the Invention [Problem to be solved by the invention]

[0006] The main objective is to provide an agent for enhancing the expression of epidermal moisturizing factors or barrier function factors. [Means for solving the problem]

[0007] As a result of intensive research aimed at solving the above-mentioned problems, the present inventors have discovered a new use for an extract of Albizia julibrissin (Silboda julibrissin) bark, namely, that the extract of Albizia julibrissin (Silboda julibrissin) bark can enhance the expression of moisturizing factors or barrier function factors in the epidermis. Based on this finding, the present inventors have conducted further research and have completed the present invention.

[0008] The present invention includes the following aspects. Section 1. An agent for enhancing the expression of moisturizing factors or barrier function factors in the epidermis, containing an extract from the bark of the Albizia julibrissin. Section 2. Item 2. The expression enhancer according to Item 1, wherein the moisturizing factor or barrier function factor is an epidermal tight junction-associated factor, an epidermal differentiation-associated factor, or an aquaporin. Section 3. Item 3. The expression enhancer according to Item 1 or 2, wherein the moisturizing factor or barrier function factor is selected from the group consisting of claudin 1, desmocollin 1, integrin 2, filaggrin, keratin 1, keratin 5, keratin 10, keratin 14, loricrin, and aquaporin 3. Section 4. A composition for preventing or treating a disease or symptom associated with decreased expression of moisturizing factors or barrier function factors in the epidermis, the composition comprising an extract of Albizia julibrissin bark. Section 5. Item 5. The composition according to Item 4, wherein the disease or symptom is selected from the group consisting of skin aging symptoms, atopic dermatitis, contact dermatitis, psoriasis, ichthyosis, xerosis, acne, allergic rhinitis, and asthma. [Effects of the Invention]

[0009] According to the present invention, an agent for enhancing the expression of an epidermal moisturizing factor or barrier function factor is provided. [Brief explanation of the drawings]

[0010] [Figure 1]FIG. 1 shows an increase in the expression of epidermal tight junction-related genes when GenemClock™ (manufactured by Toyobo Co., Ltd.) was applied to a three-dimensional cultured epidermal model. [Figure 2] FIG. 1 shows an increase in the expression of epidermal differentiation-related genes when GenemClock™ (manufactured by Toyobo Co., Ltd.) was applied to a three-dimensional cultured epidermal model. [Figure 3] FIG. 1 shows an increase in aquaporin expression when GenemClock™ (manufactured by Toyobo Co., Ltd.) was applied to a three-dimensional cultured epidermal model. DETAILED DESCRIPTION OF THE INVENTION

[0011] Silk tree bark extract The albizia bark extract is not limited to an extract obtained solely from the bark of a plant of the genus Albizia, but may be an extract obtained from the whole or a part thereof (for example, branches and leaves) as long as it contains bark. The albizia plant to be subjected to extraction may be cut as is, or may be subjected to pretreatment such as drying or pulverization. In one embodiment, the albizia bark extract is preferably extracted from the branches and leaves of a plant of the genus Albizia, either raw or, if necessary, dried and pulverized.

[0012] The extraction solvent is not particularly limited and may, for example, be water or an organic solvent. Specific examples of organic solvents include alcohols such as methanol, ethanol, propanol, and butanol; polyhydric alcohols such as propylene glycol and butylene glycol; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate; linear and cyclic ethers such as tetrahydrofuran and diethyl ether; polyethers such as polyethylene glycol; hydrocarbons such as hexane, cyclohexane, and petroleum ether; aromatic hydrocarbons such as benzene and toluene; pyridines; fats, waxes, and other oils. The extraction solvent may be used alone or in combination of two or more. When two or more extraction solvents are combined, extraction may be performed using a mixed solvent containing two or more extraction solvents, or a combination of extractions using each extraction solvent (e.g., a combination of water extraction and organic solvent extraction). The extraction solvent is preferably water, alcohols, or a mixture thereof, and more preferably water (particularly hot water at 70 to 100°C).

[0013] The extraction conditions vary depending on the solvent used. For example, when extracting with water, it is preferable to use 1 to 100 parts by mass of water per part by mass of Albizia julibrissin bark, and extract at a temperature of 4 to 100°C for 10 minutes to 7 days. It is more preferable to use 5 to 20 parts by mass of water per part by mass of Albizia julibrissin bark, and extract at a temperature of 70 to 100°C for 30 minutes to 2 hours. When extracting with hot water, extraction is typically performed under heated reflux. To further increase the extraction efficiency, pressure application, stirring, ultrasonic treatment, etc. may be performed during extraction.

[0014] The Albizia julibrissin bark extract may be directly extracted, or may be extracted and then subjected to post-treatment such as dilution, concentration, drying, separation, purification, etc. Examples of separation methods include filtration, centrifugation, etc. Examples of purification methods include fractional extraction, partition between two solvents, fractional adsorption with an adsorbent, elution, chromatography, etc.

[0015] The albizia julibrissin bark extract may be in any form, for example, a solid form such as a dried powder (including those that have been subjected to processing such as pulverization after drying), a liquid form such as an extract liquid, a dispersion or solution of the dried powder, or a paste form.

[0016] The albizia julibrissin bark extract can be prepared, for example, by the method described in JP 2018-58783 A.

[0017] Epidermal moisturizing factor or barrier function factor The moisturizing factor or barrier function factor is not particularly limited as long as it is present in the epidermis. The moisturizing factor or barrier function factor may be present in any of the layers constituting the surface layer, i.e., the stratum corneum, stratum granulosum, stratum spinosum, and stratum basale. In one embodiment, the moisturizing factor or barrier function factor of the epidermis may be, for example, an epidermal tight junction-associated factor, an epidermal differentiation-associated factor, etc. Specific examples of genes related to such functions or proteins expressed thereby include claudins (e.g., claudin 1), desmocollins (e.g., desmocollin 1), integrins (e.g., integrin 2), profilaggrin, filaggrin, loricrin, involucrin, transglutaminase, keratins (e.g., keratin 1, keratin 5, keratin 10, keratin 14), and aquaporins (e.g., aquaporin 3). Among these, one selected from the group consisting of claudin 1, desmocollin 1, integrin 2, filaggrin, loricrin, keratin 1, keratin 5, keratin 10, keratin 14, and aquaporin 3 is preferred.

[0018] Expression enhancer The expression enhancer is not particularly limited, as long as it enhances the expression of the moisturizing factor or barrier function factor in the epidermis when applied to the epidermis compared to when the expression enhancer is not applied to the epidermis. The degree of expression enhancement is not particularly limited, as long as the expression level when the expression enhancer is applied to the epidermis is more than 1-fold the expression level when the expression enhancer is not applied to the epidermis, and is, for example, 1.1-fold or more, preferably 1.2-fold or more, more preferably 1.3-fold or more. The presence or absence and degree of expression enhancement can be analyzed by microarray using a three-dimensional cultured skin model.

[0019] The expression enhancer is not particularly limited as long as it contains an Albizia julibrissin bark extract. The content of the Albizia julibrissin bark extract (converted to dry mass) is not particularly limited as long as it is an effective amount for exhibiting an expression-enhancing effect, and is, for example, 0.00001% by mass (or w / v%) or more, preferably 0.00005% by mass (or w / v%) or more, 0.0001% by mass (or w / v%) or more, 0.0005% by mass (or w / v%) or more, 0.001% by mass (or w / v%) or more, 0.005% by mass (or w / v%) or more, 0.01% by mass (or w / v%) or more, or 0.05% by mass (or w / v%) or more. The content of the Albizia julibrissin bark extract may be 100% by mass or less, for example, 50% by mass (or w / v%) or less, and preferably 30% by mass (or w / v%) or less, 20% by mass (or w / v%) or less, 10% by mass (or w / v%) or less, or 5% by mass (or w / v%) or less.

[0020] The expression enhancer preferably contains a carrier in addition to the Albizia julibrissin bark extract. The carrier is not particularly limited as long as it is physiologically or pharmaceutically acceptable, and examples thereof include water; buffer solutions such as phosphate buffer, phosphate-buffered saline, HEPES buffer, Tris-HCl buffer, borate buffer, acetate buffer, and citrate buffer; higher or lower alcohols such as caprylic alcohol, lauryl alcohol, oleyl alcohol, myristyl alcohol, cetyl alcohol, cholesterol, phytosterol, cetanol, stearyl alcohol, hexyldecanol, octyldodecanol, ethanol, and isopropanol; higher fatty acids or esters thereof (e.g., alkyl esters) such as capric acid, myristic acid, palmitic acid, stearic acid, behenic acid, lanolin fatty acid, linoleic acid, linolenic acid, lauric acid, oleic acid, isostearic acid, undecylenic acid, isononanoic acid, and caproic acid; liquid paraffin, squalane, and the like. Hydrocarbons such as cellulose, microcrystalline wax, ceresin wax, paraffin wax, and petrolatum; avocado oil, almond oil, fennel oil, perilla oil, olive oil, orange oil, orange roughage oil, sesame oil, cocoa oil, chamomile oil, carrot oil, cucumber oil, macadamia nut oil, kukui nut oil, safflower oil, soybean oil, camellia oil, corn oil, rapeseed oil, persic oil, castor oil, and cottonseed oil. Examples of suitable carriers include oils and fats such as peanut oil, turtle oil, mink oil, palm oil, palm kernel oil, coconut oil, grapeseed oil, Japan wax, jojoba oil, beef tallow, beef tallow fatty acids, lard, egg yolk oil, and hardened oil; waxes such as lanolin (liquid lanolin, reduced lanolin, hard lanolin, etc.), beeswax, carnauba wax, spermaceti, candelilla wax, montan wax, and shellac wax; and silicones such as dimethylpolysiloxane and methylphenylsiloxane. These carriers may be used alone or in combination of two or more.

[0021] The expression enhancer may further contain an additive. The additive is not particularly limited as long as it is physiologically or pharmaceutically acceptable and can be appropriately selected depending on the form and mode of use. Examples include, but are not limited to, solvents, dispersants, emulsifiers, pH adjusters, thickeners, preservatives, stabilizers, surfactants, excipients, binders, disintegrants, lubricants, sweeteners, flavorings, colorants, etc. The additives can be used alone or in combination of two or more.

[0022] The expression enhancer may be in the form of, for example, a medicine, a reagent, a food, a cosmetic (including quasi-drugs), or a composition for use therein. In the case of medicines, the enhancer may be in the form of tablets, granules, powders, pills, capsules, liquids, suspensions, elixirs, emulsions, ointments, injections, patches, etc. In the case of foods, the enhancer may be in the form of beverages such as soft drinks, carbonated drinks, nutritional drinks, fruit drinks, and lactic acid drinks; frozen desserts such as ice cream, ice sherbet, and shaved ice; noodles such as soba, udon, vermicelli, Chinese noodles, and instant noodles; sweets such as candy, candy, gum, chocolate, jelly, jam, cream, snacks, baked goods, and bread; processed seafood and livestock foods such as kamaboko, ham, and sausage; dairy products such as powdered milk, processed milk, and fermented milk; oil- and fat-processed foods such as margarine, mayonnaise, shortening, whipped cream, and dressing; condiments such as sauces and dressings; and retort pouch foods. Furthermore, the cosmetic product may be a health food, functional food, food for specified health uses, food with functional claims, nutritional supplement, supplement, etc. In the case of cosmetics, the cosmetic product may be a basic cosmetic product such as a lotion, emulsion, gel, cream, ointment, lotion, oil, pack, etc.; a facial cleanser; a skin cleanser; a hair cosmetic product such as shampoo, rinse, hair treatment, hair cream, hair styling agent, perm agent, hair tonic, hair dye, hair growth tonic, etc.; a makeup cosmetic product such as foundation, face powder, powder, lipstick, blush, eye shadow, eyeliner, mascara, etc.; a finishing cosmetic product such as nail polish, etc.; a sunscreen, etc.

[0023] When the expression enhancer is in the form of a product such as a food product, the product itself or its container or packaging may be labeled, for example, that the product is used to enhance the expression of epidermal moisturizing factors or barrier function factors (particularly, epidermal tight junction-associated factors, epidermal differentiation-associated factors, and aquaporins), that the product is used to prevent, improve, alleviate, delay progression, or inhibit progression of symptoms associated with decreased expression of epidermal moisturizing factors or barrier function factors (particularly, epidermal tight junction-associated factors, epidermal differentiation-associated factors, and aquaporins), or a similar label regarding the manner of use, and the product instructions may include a description corresponding to the above-mentioned labeling.

[0024] The expression enhancer can be suitably used for the prevention or treatment (including improvement, alleviation, delay of progression, inhibition of progression, etc.) of diseases or symptoms associated with decreased expression of epidermal moisturizing factors or barrier function factors (particularly, epidermal tight junction-associated factors, epidermal differentiation-associated factors, and aquaporins), or for application to subjects with decreased expression of epidermal moisturizing factors or barrier function factors (particularly, epidermal tight junction-associated factors, epidermal differentiation-associated factors, and aquaporins). Such diseases or symptoms are preferably diseases or symptoms associated with decreased expression of one selected from the group consisting of claudin 1, desmocollin 1, integrin 2, filaggrin, loricrin, keratin 1, keratin 5, keratin 10, keratin 14, and aquaporin 3. Specific examples include skin aging symptoms such as rough skin and dry skin; skin diseases such as atopic dermatitis, contact dermatitis, psoriasis, ichthyosis, xerosis, and acne; allergic rhinitis; and asthma.

[0025] The method of application of the expression enhancer is not particularly limited as long as it exerts an expression-enhancing effect, and may be oral or parenteral administration. The application method is preferably topical administration, such as direct application to the skin, spraying, injection, or atomization.

[0026] The present invention also encompasses compositions containing an extract of Albizia julibrissin bark for preventing or treating diseases or symptoms associated with decreased expression of epidermal moisturizing factors or barrier function factors (particularly, epidermal tight junction-associated factors, epidermal differentiation-associated factors, and aquaporins), or for application to subjects with decreased expression of epidermal moisturizing factors or barrier function factors (particularly, epidermal tight junction-associated factors, epidermal differentiation-associated factors, and aquaporins).The composition of the composition is the same as that described for the expression enhancer. [Example]

[0027] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples.

[0028] A three-dimensional cultured epidermal model (SkinEthic™ RHE, hereafter referred to as "RHE") was acclimated overnight in growth medium. After acclimation, the RHE was transferred to a 6-well plate containing 1 mL of growth medium, and 100 μL of a test sample ("Genemclock™" manufactured by Toyobo Co., Ltd., Albizia julibrissin bark extract) was applied to the stratum corneum side of the RHE. After 24 hours of culture, the test sample was removed and the RHE was washed with PBS(-).

[0029] The viability of RHE cells before RNA extraction was assessed using the Alamar Blue method. Maintenance medium containing 10% Alamar Blue reagent was dispensed into a 12-well plate. RHE cells were transferred to the plate and cultured for 2 hours. The fluorescence intensity (Ex. / Em. = 544 nm / 590 nm) of the culture supernatant was measured. Cell viability was calculated as an index (%) relative to the fluorescence intensity of the untreated group.

[0030] After measuring the fluorescence intensity, the RHE cells were punched out of the transwell and immersed in QIAzol™ Lysis reagent (QIAGEN) for frozen storage at -80°C. High-concentration conditions were selected to achieve a cell viability of 90% or higher, and the selected RHE cells were disrupted using a Tissue Lyser (QIAGEN). RNA was purified from the disrupted solution using the miRNeasy™ Mini Kit (QIAGEN). DNA microarray analysis was performed on the recovered RNA using an mRNA expression analysis chip. The results were analyzed, and the results of various gene expression analyses were expressed as ratios with the corrected value of the control (PBS(-) applied) set at 1, and significance was tested using a Student's t-test.

[0031] The results are shown in Figures 1 to 3. As shown in Figures 1 to 3, when GenemClock (trademark) was applied to RHE, enhanced expression of epidermal tight junction-associated factors, epidermal differentiation-associated factors, and aquaporins was observed.

Claims

[Claim 1] An expression enhancer for an epidermal moisturizing factor or barrier function factor, comprising an extract of Albizia julibrissin bark, wherein the moisturizing factor or barrier function factor is selected from the group consisting of claudin 1, desmocollin 1, integrin 2, filaggrin, keratin 1, keratin 5, keratin 10, keratin 14, loricrin, and aquaporin 3.

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