Periostin synthesis inhibitor

Abstract

To provide a functional material derived from naturally derived components and exhibiting a novel periostin synthesis inhibitory effect.SOLUTION: A periostin synthesis inhibitor comprising, as an active ingredient, a hydrolysate obtained through proteolytic enzyme treatment of seeds of white mustard, a plant of the genus Brassica in the family Brassicaceae, or an extract thereof.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a periostin synthesis inhibitor containing a hydrolysate of white mustard seed extract as an active ingredient. [Background technology]

[0002] In recent years, research into various allergic diseases has progressed, and a protein called periostin, which is expressed at the lesions of allergic diseases, has attracted attention. It is known that when periostin is expressed in fibroblasts due to the influence of type 2 cytokines such as IL-4 and IL-13, it reduces the expression of filaggrin via IL-24, thereby reducing the skin barrier function. [Prior art documents] [Non-patent literature]

[0003] [Non-Patent Document 1] Takeshi Nakahara et al., Western Japan Skin, Vol. 85, No. 1, 2023 DISCLOSURE OF THE INVENTION [Problem to be solved by the invention]

[0004] In view of the prior art, the present inventors conducted extensive research to find a naturally occurring component that inhibits periostin synthesis. As a result, they found that a hydrolysate obtained by hydrolyzing an extract of seeds of white mustard, a plant of the genus Brassica in the Brassicaceae family, with a protease has the effect of inhibiting periostin synthesis. [Means for solving the problem]

[0005] The present invention relates to a periostin synthesis inhibitor containing, as an active ingredient, a protease hydrolysate of white mustard seed extract. [Effects of the Invention]

[0006] The present invention provides a periostin synthesis inhibitor containing, as an active ingredient, a protease hydrolysate of white mustard seed extract. The present invention can provide a preparation that prevents damage to the barrier function due to inflammation and the like and maintains healthy skin barrier function by suppressing periostin synthesis in skin cells. DETAILED DESCRIPTION OF THE INVENTION

[0007] Preferred embodiments of the present invention will now be described in detail. The present invention relates to a periostin synthesis inhibitor containing, as an active ingredient, a hydrolysate obtained by protease hydrolysis of an extract of seeds of white mustard (Brassica alba), a plant of the genus Brassica in the family Brassicaceae (herbal medicine name: Hakugaishi "white mustard").

[0008] First, the extract can be prepared by washing the white mustard seeds, which are the part to be extracted, with water, drying them, shredding them, or crushing them, if necessary, and then contacting them with an extraction solvent using a conventional method such as immersion, countercurrent extraction, or supercritical extraction.

[0009] Examples of extraction solvents include water; lower alcohols such as ethanol; polyhydric alcohols such as propanediol, pentylene glycol, 1,3-butylene glycol, and glycerin; esters such as ethyl acetate, butyl acetate, and methyl propionate; and hydrocarbon solvents such as n-hexane, and these can be used alone or in combination.

[0010] Among these extraction solvents, the use of water, a mixed solvent of water and a lower alcohol, or a mixed solvent of water and a polyhydric alcohol is preferred in the present invention because of the ease of transition to the hydrolysis treatment step described below when the extract is subjected to said treatment and the wide range of applications to formulations. When a mixed solvent is used, the ratio of the lower alcohol or polyhydric alcohol to water is generally 70% by weight or less, preferably 50% by weight or less.

[0011] There are no particular limitations on the pH, temperature and time required for preparing the extract, but it is preferable to select them appropriately depending on the part of the plant to be extracted.

[0012] The pH of the extract during preparation is generally preferably in the range of 3 to 9. If necessary, the pH may be adjusted to the desired level by adding an alkalinity adjuster such as sodium hydroxide, sodium carbonate, or potassium hydroxide, or an acidity adjuster such as citric acid, hydrochloric acid, phosphoric acid, or sulfuric acid to the extraction solvent. The extraction conditions, such as extraction temperature and extraction time, vary depending on the type and pH of the solvent used, as well as the degree of chopping, dryness, and particle size of the extraction material. For example, in the case of a maceration method using water as the extraction solvent, the extraction temperature is generally in the range of 1 to 90°C, preferably 20 to 60°C, and the extraction time is, for example, in the range of 0.5 to 72 hours, preferably 1 to 24 hours, at room temperature.

[0013] In the present invention, the extract obtained by the above extraction treatment is subjected to hydrolysis treatment before or after extraction, or in parallel with extraction. As the hydrolysis treatment, hydrolysis treatment using an enzyme is preferred, and protease is more preferred. The protease may be used alone or in combination of two or more enzymes.

[0014] Examples of protease include actinases such as actinase, pepsins such as pepsin, trypsins such as trypsin and chymotrypsin, papains such as papain and chymopapain, peptidases such as glycylglycine peptidase, carboxypeptidase, and aminopeptidase, bromelain, and conjugated proteases derived from microorganisms [e.g., Neurase (manufactured by Amano Enzyme Inc.)]. When a protease is used, any of the above enzymes may be used alone, or a plurality of enzymes may be used in combination.

[0015] Enzymatic hydrolysis is carried out by adding one or more of the enzymes listed above to the plant extract solution and allowing the enzyme to react under conditions near the optimum pH and temperature for the enzyme used. When using a combination of two or more enzymes, the enzymes may be allowed to act simultaneously depending on their properties. The amount of enzyme used is preferably in the range of 0.001 to 50 parts by weight, more preferably 0.1 to 10 parts by weight, per 100 parts by weight of the solids content of the plant extract solution. The time for the enzyme treatment varies depending on the type of enzyme used, but is generally in the range of 0.5 to 24 hours, and preferably 1 to 6 hours.

[0016] After hydrolysis using the enzymes described above is completed, the enzymes are inactivated by an appropriate method, such as heating the enzyme-treated solution to 80° C. or higher, to obtain an enzyme-treated decomposition product solution.

[0017] The hydrolysate prepared as described above may be blended as it is into topical skin preparations (cosmetics, quasi-drugs, or topical pharmaceuticals) after generally adjusting the pH to 4 to 8, or may be used at a desired concentration by concentrating under reduced pressure, etc. Alternatively, it may be dried by a conventional method such as spray drying.

[0018] The hydrolysate prepared as described above may be stored in a refrigerator for a certain period of time to enhance storage stability, and the supernatant may then be used.

[0019] The hydrolysate according to the present invention can be incorporated into external skin preparations (cosmetics, quasi-drugs, or external pharmaceuticals). Examples of external skin preparations include emulsions, creams, lotions, essences, packs, masks, lipsticks, foundations, liquid foundations, makeup press powders, blushers, face powders, face washes, body shampoos, scalp and hair shampoos, and bath additives, but the present invention is not limited to these.

[0020] The amount of the hydrolyzate according to the present invention can be adjusted appropriately depending on the formulation to be used, and the solid content is, for example, in the range of 0.002 to 1.0 wt % (solid content weight %, the same applies hereinafter) for skin care cosmetics, 0.002 to 1.0 wt % for makeup cosmetics, 0.002 to 10.0 wt % for cleansing cosmetics, and 0.0001 to 5.0 wt % for hair cosmetics as the solid content of the composition.

[0021] When the hydrolysate according to the present invention is incorporated into an external skin preparation, in addition to the above-mentioned composition as the essential component, optional components generally used in external skin preparations can be incorporated. Suitable examples of optional components include hydrocarbons such as squalane, petrolatum, and microcrystalline wax; esters such as jojoba oil, carnauba wax, and octyldodecyl oleate; triglycerides such as olive oil, beef tallow, and coconut oil; fatty acids such as stearic acid, oleic acid, and retinoic acid; higher alcohols such as oleyl alcohol, stearyl alcohol, and octyldodecanol; anionic surfactants such as sulfosuccinate esters and polyoxyethylene alkyl sulfate sodium; amphoteric surfactants such as alkyl betaine salts; cationic surfactants such as dialkylammonium salts; nonionic surfactants such as sorbitan fatty acid esters, fatty acid monoglycerides, polyoxyethylene adducts thereof, polyoxyethylene alkyl ethers, and polyoxyethylene fatty acid esters; polyhydric alcohols such as polyethylene glycol, glycerin, and 1,3-butanediol; thickeners / gelling agents; antioxidants; ultraviolet protection agents such as ultraviolet absorbers and ultraviolet scattering agents; colorants; preservatives; pH adjusters; powders; and various active ingredients.

[0022] Examples of production and efficacy evaluation tests for the hydrolysate, which is the active ingredient of the present invention, are described below, but the present invention is not limited thereto. Furthermore, all "parts" below mean parts by weight, and all "%" means % by weight.

[0023] Production Example 1. Preparation of hydrolysate (1) 50 g of ground white mustard seeds (white mustard) was mixed with 1000 g of purified water, and the mixture was extracted at 40°C for 1 hour, followed by filtration to obtain 805 g of a pale yellow, transparent white mustard extract solution (solid concentration: 1.22 wt%). Next, 0.05 g of actinase was added as a hydrolytic enzyme to 500 g of the resulting extract solution, and hydrolysis was carried out at 40°C for 2 hours. The enzyme was then inactivated by heating at 85°C for 1 hour, and the mixture was filtered to obtain 462 g of a pale yellow, transparent white mustard extract hydrolysate solution (solid concentration: 1.01 wt%).

[0024] In Production Example 1, other proteases, such as papain, may be used as the hydrolase instead of actinase.

[0025] Test Example 1: Evaluation of periostin synthesis inhibitory effect Normal epidermal keratinocytes (NHEK) were plated in a 24-well plate at 5.0 x 10 4 The cells were seeded at 1000 cells / well and cultured at 37°C under 5% CO2 and saturated steam for 24 hours. Next, the hydrolysate of Production Example 1 was added as a sample solution, and the culture was continued for another 24 hours. Here, the sample solution was set at two concentrations, with final concentrations of 1.0% and 2.0% relative to the total volume of the culture medium. In addition, as a comparative control, a test group (control group) was set in which a culture medium containing a 30% BG solution of the same concentration was added instead of the sample. Next, the culture medium was removed, washed once with PBS(-), and then PBS(-) was added. Approximately 50 mJ / cm was irradiated from the bottom of the culture vessel using a UV-B lamp (Philips TL20W / 12RS). 2 The supernatant was then replaced with HuMedia KB2 medium, and the culture was continued. After 24 hours of culture, the medium was collected. Meanwhile, normal fibroblast NB1RGB cells were cultured in a 96-well plate at 1.0 × 10 4The fibroblasts were seeded at 100 cells / well and cultured for 24 hours at 37°C under 5% CO2 and saturated water vapor. The culture supernatant of the epidermal cells collected above was then added as a sample, and culture was continued. After 48 hours of culture, the medium was collected. The periostin content in the culture supernatant was quantified using a periostin ELISA kit (Abcam ab213816) for this fibroblast culture supernatant as a sample. The results are expressed as a relative value, with the periostin content in the group to which the culture supernatant of UV-irradiated epidermal cells was added being set at 100.

[0026] The results of Test Example 1 are shown in Table 1. [Table 1] JPEG2025133358000001.jpg52133

[0027] As shown in Table 1, it was first confirmed that adding the supernatant of UV-irradiated epidermal cells to a fibroblast culture system increased the amount of periostin synthesized, and that the hydrolysate of Production Example 1 of the present invention has the effect of suppressing periostin synthesis. Thus, by incorporating the hydrolysate of the present invention as an active ingredient, a topical skin preparation that maintains the skin barrier function in a healthy state can be provided.

[0028] Prescription example 1. Lotion [Ingredients] Part Squalane 0.20 Polyoxyethylene (5.5) Cetyl Alcohol 5.00 Tocopherol acetate 0.02 Dipotassium glycyrrhizinate 0.10 1,3-butylene glycol 5.00 Hydrolyzate of Production Example 1 1.00 Niacinamide 5.00 Sodium citrate 0.20 Pentanediol 0.10 Purified water (enough to make the total volume 100 parts)

[0029] Prescription example 2: Lotion [Ingredients] Part Squalane 0.20 Polyoxyethylene (5.5) Cetyl Alcohol 5.00 Tocopherol acetate 0.02 Tranexamic acid 1.00 1,3-butylene glycol 5.00 Hydrolyzate of Production Example 1 1.00 Niacinamide 5.00 Sodium citrate 0.20 Pentanediol 0.10 Purified water (enough to make the total volume 100 parts)

[0030] Prescription example 3: Lotion [Ingredients] Part Squalane 0.20 Polyoxyethylene (5.5) Cetyl Alcohol 5.00 Tocopherol acetate 0.02 D-Pantothenyl alcohol 0.10 1,3-butylene glycol 5.00 Hydrolyzate of Production Example 1 1.00 Niacinamide 5.00 Sodium citrate 0.20 Pentanediol 0.10 Purified water (enough to make the total volume 100 parts)

[0031] Prescription example 4. Cream [Ingredients] Part Stearic acid 1.00 Glyceryl monostearate 0.50 Squalane 15.00 Sorbitan Monostearate 2.00 Polyoxyethylene (20) Sorbitan Monostearate 2.00 Carboxyvinyl polymer 0.10 Hydrolyzate of Production Example 1 1.00 Ascorbic acid derivative (ascorbic acid glucoside) 2.00 Methylparaben 0.10 Potassium hydroxide (appropriate amount) Purified water (enough to make the total volume 100 parts)

[0032] Prescription example 5. Cream [Ingredients] Part Stearic acid 1.00 Glyceryl monostearate 0.50 Squalane 15.00 Sorbitan Monostearate 2.00 Polyoxyethylene (20) Sorbitan Monostearate 2.00 Carboxyvinyl polymer 0.10 Hydrolyzate of Production Example 1 1.00 Ascorbic acid derivative (ascorbic acid glucoside) 2.00 Arbutin 3.00 Methylparaben 0.10 Potassium hydroxide (appropriate amount) Purified water (enough to make the total volume 100 parts)

Claims

[Claim 1] A periostin synthesis inhibitor containing, as an active ingredient, a protease hydrolyzate of seeds or an extract of white mustard, which is a plant of the genus Brassica in the Brassicaceae family.

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