Epidermal barrier function improving agent, aromatic hydrocarbon receptor activator

Abstract

The present invention aims to provide an epidermal barrier function improving agent and an aromatic hydrocarbon receptor activator containing Artemisia princeps extract and / or vitamin K as active ingredients. [Solution] The present invention provides an epidermal barrier function improving agent and an aromatic hydrocarbon receptor activator containing Artemisia princeps extract and / or vitamin K as active ingredients.

Description

Technical Field

[0001] The present invention relates to an epidermal barrier function improver and an aromatic hydrocarbon receptor activator containing Artemisia princeps extract and / or vitamin K as active ingredients.

Background Art

[0002] The aromatic hydrocarbon receptor (AhR) exists in the cytoplasm and translocates to the nucleus upon binding to a ligand, inducing the transcription of response genes such as the drug-metabolizing enzyme CYP1A1 (CytochromeP450 family 1 subfamily A member 1). In recent years, in atopic dermatitis, it has been reported that tapinarof, an AhR regulator, has an effect of improving the decline in skin barrier function (Non-Patent Document 2).

Prior Art Documents

Non-Patent Documents

[0003]

Non-Patent Document 1

Non-Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0004] An object of the present invention is to provide an epidermal barrier function improver and an aromatic hydrocarbon receptor activator containing Artemisia princeps extract and / or vitamin K as active ingredients.

Means for Solving the Problems

[0005] The means for solving the problems of the present invention are as follows. (1) An epidermal barrier function improving agent containing Artemisia princeps extract and / or vitamin K as active ingredients. (2) An aromatic hydrocarbon receptor activator containing Artemisia princeps extract and / or vitamin K as active ingredients. [Effects of the Invention]

[0006] The present invention provides an epidermal barrier function improving agent and aromatic hydrocarbon receptor activator containing Artemisia japonica extract and / or vitamin K as active ingredients, exhibiting high epidermal barrier function improving effects, aromatic hydrocarbon receptor activation effects, FLG expression promoting effects, IVL expression promoting effects, OVOL1 expression promoting effects, IL33 expression suppression effects, and IL37 expression promoting effects in epidermal keratinocytes. [Modes for carrying out the invention]

[0007] The following describes embodiments for carrying out the present invention.

[0008] CYP1A1 is an abbreviation for Cytochrome P450 family 1 subfamily A member 1, and is an AhR-responsive gene involved in drug metabolism.

[0009] FLG (Filaggrin) is a gene involved in the skin's barrier function, and it has been reported that FLG expression levels are decreased in both lesional and non-lesional areas of atopic dermatitis.

[0010] IVL (Involcrin) is a gene involved in the skin's barrier function.

[0011] IL33 (Interleukin-33) is a gene that encodes an inflammatory cytokine and induces type 2 cytokines [IL4 (Interleukin-4), IL13 (Interleukin-13)] which play a central role in the pathogenesis of atopic dermatitis.

[0012] IL37 (Interleukin-37) is an anti-inflammatory cytokine that suppresses the production of inflammatory cytokines.

[0013] [Artemisia princeps extract] The Artemisia montana (Nakai) Pamp. used in this invention is a plant belonging to the Artemisia genus of the Asteraceae family, and is also known as Artemisia montana or Artemisia japonica.

[0014] Parts of Artemisia princeps that can be used for extraction include, for example, the roots, leaves, stems, flowers, buds, and the entire above-ground plant, but the leaves are preferred in terms of effectiveness.

[0015] In this invention, Artemisia princeps may be used for extraction in its raw state, but considering the extraction efficiency, it is preferable to perform processing such as finely chopping, drying, or grinding before extraction. Extraction is carried out by immersion in the extraction solvent. Stirring or homogenization in the extraction solvent may be performed to increase the extraction efficiency. The extraction temperature should be between approximately 5°C and below the boiling point of the extraction solvent. The extraction time will vary depending on the type of extraction solvent and the extraction temperature, but it should be between approximately 4 hours and 30 days.

[0016] As extraction solvents, organic solvents such as lower alcohols like methanol, ethanol, propanol, and isopropanol; polyhydric alcohols like 1,3-butylene glycol, propylene glycol, dipropylene glycol, and glycerin; ethers like ethyl ether and propyl ether; esters like ethyl acetate and butyl acetate; and ketones like acetone and ethyl methyl ketone can be used, and one or more of these can be selected and used. Alternatively, physiological saline, phosphate buffer, or phosphate-buffered saline may also be used.

[0017] The extract of Artemisia princeps Pamp. with the above solvent can be used as it is, but it can also be concentrated and dried, then redissolved in water or a polar solvent, or subjected to purification treatments such as decolorization, deodorization, and desalting within a range that does not impair its skin physiological function improvement effect, or fractionated by column chromatography before use. Also, for storage, it can be freeze-dried after purification treatment and dissolved in a solvent when in use. It can also be encapsulated in vesicles such as liposomes or microcapsules for use.

[0018] In the present invention, it is preferable to use an extract obtained from wild Artemisia princeps Pamp. collected from Yuzawa, Shimo-cho, Shibetsu, Hokkaido from May to August.

[0019] [Vitamin K] Vitamin K is an essential component for the activation of vitamin K-dependent proteins, is related to blood coagulation and tissue calcification in the animal body, and it is known that a deficiency of vitamin K leads to a bleeding tendency. In terms of chemical structure, vitamin K is a 3-derivative of 2-methyl-1,4-naphthoquinone. Naturally, it contains K1 and K2 (menaquinone). Menaquinone (MK) contains a number of compounds with different lengths and modifications of the isoprenoid side chain, and all these vitamin Ks can be used.

[0020] The Artemisia princeps Pamp. extract and / or vitamin K of the present invention exhibit a high epidermal barrier function improvement effect, aromatic hydrocarbon receptor activation effect, FLG expression promotion effect, IVL expression promotion effect, OVOL1 expression promotion effect, IL33 expression inhibition effect, and IL37 expression promotion effect in epidermal keratinocytes, and are useful as an epidermal barrier function improver, aromatic hydrocarbon receptor activator, FLG expression promoter, IVL expression promoter, OVOL1 expression promoter, IL33 expression inhibitor, and IL37 expression promoter.

[0021] Each agent containing the Artemisia princeps Pamp. extract and / or vitamin K of the present invention as an active ingredient can be used alone, but can also be formulated into various compositions such as pharmaceuticals, quasi-drugs, foods and drinks, and cosmetics.

[0022] Each agent containing the Artemisia princeps Pamp. extract and / or vitamin K of the present invention as an active ingredient is not restricted in any way regarding its form and the presence or absence of other components. Regarding the form, any form such as liquid, paste, gel, solid, powder, etc. can be selected according to its use, etc., and it can optionally contain a vehicle (excipient), solvent, and other general additives (antioxidants, colorants, dispersants, etc.) necessary for taking that form.

[0023] The composition containing each agent containing the Artemisia princeps Pamp. extract and / or vitamin K of the present invention as an active ingredient can take any dosage form. When the composition is a skin cosmetic, hair cosmetic, cleansing agent, etc., it can be provided as a solubilized system such as a lotion, a dispersion system such as calamine lotion, or an emulsified system such as a cream or emulsion. Furthermore, it can be provided in various dosage forms such as an aerosol form filled with an aerosol agent, an ointment, a patch, etc.

[0024] Specifically, various cosmetics such as emulsions, creams, lotions, skin tonics, packs, beauty essences, cleansing agents, makeup cosmetics, etc.; cosmetics, quasi-drugs, and external medicines in various forms such as liquids, ointments, powders, granules, aerosol agents, patches, patches, etc. can be exemplified.

[0025] In addition, in the case of oral pharmaceuticals, etc., it can be made into general dosage forms such as liquids such as drinkable agents and drip agents, solid agents such as gums and candies, capsules, powders, granules, tablets, etc.

[0026] In addition to the above, compositions containing each agent of the present invention that contains Artemisia japonica extract and / or vitamin K as an active ingredient may also contain, depending on the intended use and requirements, any ingredients commonly used in pharmaceuticals, quasi-drugs, skin cosmetics, hair cosmetics, cleansers, and oral pharmaceuticals, such as water, oily components, humectants, powders, pigments, emulsifiers, solubilizers, gelling agents, cleansing agents, UV absorbers, anti-inflammatory agents, thickeners, pH adjusters, chelating agents, drugs (pharmacoactive ingredients), fragrances, resins, antibacterial and antifungal agents, antioxidants, alcohols, etc. Furthermore, within the limits that do not impair the effects of the present invention, it is also possible to use other humectants, anti-aging agents, whitening agents, antioxidants, slimming agents, or various plants / fungi or their extracts in combination. [Examples]

[0027] The present invention will be specifically described below with reference to examples, but this will not limit the scope of the present invention.

[0028] [Artemisia princeps extract (1)] Leaves of Artemisia princeps harvested in July in Yunosawa, Mashike-cho, Hokkaido were dried and immersed in 20 times the mass of a 50% ethanol aqueous solution. The liquid was collected after filtration, and the solvent was removed by distillation. The obtained extract was dissolved in a 50% 1,3-butylene glycol aqueous solution to a concentration of 0.6% by mass of extract, and then filtered again to prepare Artemisia princeps extract (1).

[0029] [test] Normal human epidermal keratinocytes 5 × 10 5Cells were seeded in 24-well plates at a density of cells / well and cultured in Humedia-KG2 medium for 24 hours. The medium was then replaced with Humedia-KG2 medium supplemented with a predetermined amount of Artemisia princeps extract or vitamin K, and cultured for 48 hours in a 37°C, 5% CO2 incubator. RNA was extracted from the collected cells using a commercially available RNA extraction kit (Maxwell registered trademark RSC simplyRNE Cell Kit), and after cDNA synthesis, gene expression was confirmed by real-time PCR using the CyberGreen method with the following primers. RPLP0 was used as the internal standard. mRNA expression levels are shown as relative values, with the expression level without Artemisia princeps extract or vitamin K set to 1. Phylloquinone (reagent) was used as vitamin K.

[0030] [Table 1]

[0031] [Table 2]

[0032] As shown in Table 2, adding 0.25% of Artemisia japonica extract (1) to the test significantly increased the expression level of CYP1A1, an indicator of AhR activation.

[0033] Furthermore, as shown in Table 2, adding 0.25% of Artemisia japonica extract (1) to the test significantly increased the expression levels of FLG, IVL, and OVOL1, which are involved in skin barrier function.

[0034] [Table 3]

[0035] As shown in Table 3, when the test was conducted with the addition of 0.125% of Artemisia japonica extract (1), a significant decrease in the inflammatory cytokine IL33 and a significant increase in the anti-inflammatory cytokine IL37 were observed.

[0036] [Table 4]

[0037] As shown in Table 4, adding 10 μM of vitamin K to the test significantly increased the expression level of CYP1A1, an indicator of AhR activation.

[0038] As shown in Table 4, adding 10 μM of vitamin K to the test significantly increased FLG and IVL, which are involved in skin barrier function.

[0039] [Artemisia japonica extract (2)~(6)] Leaves of Artemisia princeps harvested in July in Yunosawa, Mashike-cho, Hokkaido were dried, immersed in 20 times the volume of water or an aqueous ethanol solution, filtered, and the resulting liquid was collected and freeze-dried. The obtained extracts were dissolved in water or DMSO to an extract purity of 0.6 w / v%, and these were designated as Artemisia princeps extracts (2) to (6). These were then used in tests at an additive concentration of 0.0015% extract purity. As shown in Table 5, a correlation was observed between ethanol concentration and CYP1A1 expression.

[0040] [Table 5]

Claims

1. An epidermal barrier function improving agent containing Artemisia princeps extract and / or vitamin K as active ingredients.

2. An aromatic hydrocarbon receptor activator containing Artemisia princeps extract and / or vitamin K as an active ingredient.

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