Agent for improving skin microbiota and cosmetic for improving skin microbiota

Abstract

To provide a novel agent for improving skin microbiota.SOLUTION: Provided is an agent for improving skin microbiota, comprising enzyme-treated royal jelly as an active ingredient.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a skin bacterial flora-improving agent and a cosmetic for improving skin bacterial flora. [Background technology]

[0002] The skin flora (also called skin resident bacteria) that exists on human skin is made up of hundreds of different types of bacteria, with good bacteria that contribute to health coexisting with bad bacteria that are involved in disease. This state of coexistence is generally referred to as the diversity of the skin flora. It has been reported that the skin flora differs depending on the condition of the skin. For example, in atopic dermatitis, it is said that the diversity of the skin flora decreases and the proportion of Staphylococcus aureus (bad bacteria) increases.

[0003] Patent Document 1 discloses a skin bacterial flora improver that is characterized by containing fermented milk whey obtained by fermenting milk with Lactobacillus helveticus. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] JP 2022-55482 A Summary of the Invention [Problem to be solved by the invention]

[0005] An object of the present invention is to provide a novel skin bacterial flora improving agent. [Means for solving the problem]

[0006] Means of the Invention The present inventors have conducted intensive research to achieve the above object and have found that enzyme-treated royal jelly has an excellent effect of improving the skin bacterial flora, thereby completing the present invention.

[0007] That is, the present invention relates to the following [1] to [7]. [1] A skin flora improver that contains enzyme-treated royal jelly as an active ingredient. [2] The skin bacterial flora improving agent described in [1], which is used to reduce the proportion of at least one species selected from the group consisting of Cutibacterium and Staphylococcus in the skin bacterial flora. [3] The skin flora improving agent according to [2], wherein the Cutibacterium genus bacterium is Propionibacterium acnes and the Staphylococcus genus bacterium is Staphylococcus aureus. [4] The skin bacterial flora improving agent according to any one of [1] to [3], which is used to improve the diversity of skin bacterial flora. [5] The skin bacterial flora improving agent according to any one of [1] to [4], wherein the enzyme-treated royal jelly is treated with a peptidase having endopeptidase activity and exopeptidase activity. [6] The skin bacterial flora improving agent described in any one of [1] to [5], which is for external use on the skin. [7] A cosmetic for improving skin flora, comprising the skin flora improving agent according to any one of [1] to [6]. Effect of the Invention

[0008] The present invention provides a novel skin bacterial flora improving agent. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] Hereinafter, the embodiment for carrying out the present invention will be described in detail, however, the present invention is not limited to the following embodiment.

[0010] In this specification, the term "comprise" includes the meanings of "essentially consist of" and "consist only of."

[0011] The skin bacterial flora improving agent of the present invention contains enzyme-treated royal jelly as an active ingredient. The skin bacterial flora improving agent of the present invention contains enzyme-treated royal jelly as an active ingredient, and is thereby capable of improving the skin bacterial flora.

[0012] Improvement of the skin bacterial flora includes, for example, increasing the diversity, maintaining or increasing the proportion (occupancy rate) of beneficial bacteria, or decreasing the proportion of harmful bacteria in the skin bacterial flora.

[0013] The improvement of the diversity of the skin microbiota means that the kinds of bacteria contained in the skin microbiota increase. The diversity of the skin microbiota can be indicated by, for example, the Shannon diversity index. The higher the diversity, the higher the Shannon diversity index. In addition, the more kinds there are and the higher the evenness of each kind, the higher the Shannon diversity index.

[0014] The skin microbiota improving agent of the present invention can reduce the ratio of bacteria of the genus Cutibacterium and bacteria of the genus Staphylococcus, for example. The bacteria of the genus Cutibacterium may be Cutibacterium acnes. The bacteria of the genus Staphylococcus may be Staphylococcus aureus.

[0015] According to a report by the Japan Allergy Instructor Certification Board, atopic dermatitis patients have low diversity of skin flora and a high proportion of Staphylococcus aureus. The skin flora improving agent of the present invention can improve the diversity of skin flora and reduce the proportion of Staphylococcus aureus, so it is expected that atopic dermatitis can be improved by using the skin flora improving agent of the present invention.

[0016] Although acne bacteria itself is not harmful, it is said that it proliferates and is involved in the occurrence of acne depending on the skin condition. Since acne bacteria can be reduced by using the skin flora improving agent of the present invention, it is considered that the use of the skin flora improving agent of the present invention is effective in preventing or improving acne.

[0017] The skin microbiota improving agent of the present invention can be suitably used for healthy skin (skin without a disease). The skin microbiota improving agent of the present invention may be used for, for example, the skin of a person with atopic dermatitis or the skin of a person with acne.

[0018] The skin flora improving agent of the present invention also encompasses the meaning of an additive that imparts a skin flora improving effect.

[0019] Royal jelly is a milky white jelly-like substance made by mixing secretions secreted from the hypopharyngeal gland and mandibular gland by 3-12-day-old worker bees among honeybees. The main physiologically active components in royal jelly include, for example, organic acids such as saturated fatty acids and unsaturated fatty acids that are specific to royal jelly, proteins, amino acids, peptides, lipids, sugars, vitamin B, vitamins such as folic acid, nicotinic acid, and pantothenic acid, and various minerals. Royal jelly may be produced in any of European countries, Oceanian countries, America, Brazil, Japan, China, and other Asian countries.

[0020] The enzyme-treated royal jelly is royal jelly treated with a proteolytic enzyme (protease). Preferably, the enzyme-treated royal jelly is hypoallergenic, in which allergic reactions caused by proteins contained in royal jelly are suppressed by protease treatment. Therefore, the enzyme-treated royal jelly may contain organic acids such as the above-mentioned saturated fatty acids and unsaturated fatty acids, lipids, sugars, vitamins, and various minerals, in addition to the protease decomposition products of proteins contained in royal jelly.

[0021] The royal jelly used to produce the enzyme-treated royal jelly is not particularly limited, and examples thereof include raw royal jelly, royal jelly powder obtained by drying and powdering raw royal jelly, and raw royal jelly extracted with water or aqueous ethanol, etc.

[0022] Enzyme-treated royal jelly can be produced by treating royal jelly raw material with an enzyme having at least endopeptidase activity, an enzyme having at least exopeptidase activity, and / or an enzyme having both endopeptidase activity and exopeptidase activity.

[0023] Examples of proteolytic enzymes having at least endopeptidase activity include endopeptidases derived from animals (e.g., trypsin, chymotrypsin, etc.), plants (e.g., papain, etc.), and microorganisms (e.g., lactic acid bacteria, yeast, mold, Bacillus subtilis, actinomycetes, etc.).

[0024] Examples of proteolytic enzymes having at least exopeptidase activity include carboxypeptidase, aminopeptidase, exopeptidases derived from microorganisms (e.g., lactic acid bacteria, Aspergillus bacteria, Rhizopus bacteria, etc.), and pancreatin and pepsin which also have endopeptidase activity.

[0025] Among these various enzymes, preferred examples of enzymes having both exopeptidase activity and endopeptidase activity include peptidase produced by Streptomyces griseus (trade name: Actinase AS), peptidase produced by Aspergillus oryzae (trade name: Protease A, Flavorzyme, Proteax), and peptidase produced by Aspergillus melleus (trade name: Protease P). Preferred examples of enzymes having exoprotease activity include peptidase produced by Aspergillus oryzae (trade name: Umamizyme G, Promod 192P, Promod 194P, Sumizyme FLAP), peptidase produced by Aspergillus sojae (trade name: Sternzyme B15024), peptidase produced by the genus Aspergillus (trade name: Coclase P), and peptidase produced by Rhizopus oryzae (trade name: Peptidase R). Further preferred examples of enzymes having endoprotease activity include peptidase produced by Bacillus subtilis (trade name: Orientase 22BF, Nucleisin), peptidase produced by Bacillus licheniformis (trade name: Alcalase), peptidase produced by Bacillus stearothermophilus (trade name: Protease S), peptidase produced by Bacillus amyloliquefaciens (trade name: Neutrase), and peptidase produced by the genus Bacillus (trade name: Protamex).

[0026] Enzyme treatment to reduce the allergenicity of royal jelly can be carried out, for example, according to the description in JP-A-2007-295919 and JP-A-2007-295920.

[0027] The enzyme-treated royal jelly may be a filtrate obtained by filtering an enzyme-treated royal jelly solution. Filtration can be performed, for example, by suction filtration or a filter press with a particle pore size of less than 10 μm.

[0028] Commercially available enzyme-treated royal jelly may be used. A specific example of commercially available enzyme-treated royal jelly is Enzymatically Decomposed Royal Jelly King (manufactured by Yamada Bee Farm).

[0029] The content of the above-mentioned active ingredient in the skin microbiota improving agent of the present invention, in terms of solid content relative to the total amount of the skin microbiota improving agent, may be, for example, 0.1% by mass or more, 1% by mass or more, 3% by mass or more, 3.5% by mass or more, 5% by mass or more, 7% by mass or more, 10% by mass or more, 15% by mass or more, 20% by mass or more, 25% by mass or more, 30% by mass or more, 35% by mass or more, 40% by mass or more, 45% by mass or more, 50% by mass or more, 55% by mass or more, 60% by mass or more, 65% by mass or more, 70% by mass or more, 75% by mass or more, 80% by mass or more, 85% by mass or more, 90% by mass or more, 93% by mass or more, 95% by mass or more, 98% by mass or more, 99% by mass or more, or 100% by mass. The content of the above active ingredient in the skin flora improving agent is solid content based on the total amount of the skin flora improving agent, for example, 100% by mass or less, 99% by mass or less, 98% by mass or less, 95% by mass or less, 93% by mass or less, 90% by mass or less, 85% by mass or less, 80% by mass or less, 75% by mass or less, 70% by mass or less, 65 It may be less than or equal to 60 mass%, 55 mass% or less, 50 mass% or less, 45 mass% or less, 40 mass% or less, 35 mass% or less, 30 mass% or less, 25 mass% or less, 20 mass% or less, 15 mass% or less, 10 mass% or less, 8 mass% or less, 5 mass% or less, 3 mass% or less, or 1 mass% or less.

[0030] The skin flora improving agent of the present invention can be applied to the skin. The amount of the skin flora improving agent of the present invention administered to the skin may vary depending on the site and range of application, but for example, the amount of the active ingredient to be applied to the skin may be 0.01 mg to 50 mg, preferably 0.02 mg to 40 mg, or 0.02 mg to 35 mg, or 0.025 mg to 30 mg, calculated as a dry solid content.

[0031] The skin flora improving agent of the present invention can be used as an ingredient in cosmetics. A cosmetic containing the skin flora improving agent of the present invention as an ingredient can be a cosmetic for improving skin flora. The cosmetic of the present invention also includes quasi-drugs. The content of the active ingredient in the cosmetic containing the skin flora improving agent of the present invention is not particularly limited, and may be an effective amount that can achieve the dosage of parenteral administration of the agent.

[0032] A cosmetic product containing the skin flora improving agent of the present invention as an ingredient can be produced, for example, by adding the skin flora improving agent of the present invention to an intermediate product in the production process of the cosmetic product.

[0033] The pH of the skin flora improving agent of the present invention or a cosmetic product containing the skin flora improving agent may be, for example, 4.5 to 6.5, or 5.0 to 6.5.

[0034] The cosmetic product may be in any form, such as a solid, liquid, or paste. The cosmetic product may be a medicated cosmetic product (i.e., a quasi-drug). The cosmetic product includes any cosmetic product that can be applied to the skin, mucous membranes, body hair, head hair, scalp, nails, teeth, facial skin, lips, and other parts of animals (including humans).

[0035] Cosmetics can take a wide variety of formulations, including aqueous solutions, solubilized solutions, emulsions, powders, oils, gels, ointments, aerosols, water-oil two-layer systems, and water-oil-powder three-layer systems.

[0036] The use of the cosmetics is arbitrary, and examples of basic cosmetics include facial cleanser, lotion, milky lotion, cream, gel, essence, beauty serum, pack, mask, mist, UV protection cosmetics, etc.; makeup cosmetics include foundation, lipstick, blusher, eye shadow, eyeliner, mascara, etc.; nail cosmetics include nail polish, base coat, top coat, nail polish remover, etc.; and other examples include facial cleanser, (paste or liquid) dentifrice, mouth rinse, oral cosmetics, massage agents, cleansing agents, aftershave lotion, preshave lotion, shaving cream, body soap, soap, shampoo, rinse, hair treatment, hair styling agent, hair tonic agent, hair mist, hair foam, hair liquid, hair gel, hair spray, hand cream, hand soap, hair growth agent, antiperspirant, bath agent, etc.

[0037] In addition to the enzyme-treated royal jelly, the above-mentioned cosmetic product may contain, as necessary, ingredients that are typically used in cosmetics, such as alcohol, disinfectants, preservatives, surfactants, alcohols, aqueous components, water, colorants, pH adjusters, solubilizers, abrasives, foaming agents, enzymes, flavorings, chelating agents, excipients, thickeners, bases, emulsifiers, solvents, stabilizers, oils, cleaning agents (lactic acid bacteria), whitening agents, moisturizers, antioxidants, UV absorbers, powder components, colorants, and various skin nutrients.

[0038] The alcohol may be, for example, 1,3-butylene glycol, pentylene glycol, or phenoxyethanol. The total content of the alcohol in the cosmetic product may be, for example, 5% by weight or more, 7% by weight or more, 10% by weight or more, or 12% by weight or more, and may be 20% by weight or less, 18% by weight or less, or 15% by weight or less, based on the total amount of the cosmetic product.

[0039] The oil may be, for example, caprylic / capric triglyceride, etc. The content of the oil in the cosmetic product may be, for example, 0.5% by weight or more, 0.8% by weight or more, 1.0% by weight or more, 1.3% by weight or more, and 5% by weight or less, 3% by weight or less, 2% by weight or less, based on the total amount of the cosmetic product.

[0040] The thickener may be, for example, a hydrophilic thickener. The thickener may be, for example, a carbomer, etc. The content of the thickener in the cosmetic product may be, for example, 0.05% by weight or more, 0.1% by weight or more, 0.15% by weight or more, 0.2% by weight or more, and 3% by weight or less, 1% by weight or less, 0.5% by weight or less, 0.3% by weight or less, based on the total amount of the cosmetic product.

[0041] The cosmetic product containing the dermis thickness improving agent and skin texture improving agent of the present invention may further contain (acrylates / alkyl acrylate (C10-30)) crosspolymer. These are cosmetic ingredients known as thickeners and emulsion stabilizers, and are preferred because they have the effect of promoting the penetration of royal jelly into the skin.

[0042] The (acrylates / alkyl acrylate (C10-30)) crosspolymer is a copolymer of one or more monomers consisting of acrylic acid, methacrylic acid or simple esters thereof and an alkyl acrylate crosslinked with an allyl ether of sucrose or an allyl ether of pentaerythritol. Specifically, the alkyl acrylate is an alkyl acrylate having an alkyl group having 10 to 30 carbon atoms. As the (acrylates / alkyl acrylate (C10-30)) crosspolymer, a commercially available (acrylates / alkyl acrylate (C10-30)) crosspolymer (e.g., Pemulen (trademark) TR-1) may be used.

[0043] The content of the (acrylates / alkyl acrylate (C10-30)) crosspolymer in the cosmetic product may be, for example, 0.01% by mass or more, 0.1% by mass or more, 0.3% by mass or more, 0.4% by mass or more, 0.5% by mass or more, 1% by mass or more, 2% by mass or more, 3% by mass or more, 5% by mass or more, and may be 20% by mass or less, 15% by mass or less, 10% by mass or less, 8% by mass or less, 5% by mass or less, 3% by mass or less, 2% by mass or less, 1% by mass or less, 0.5% by mass or less, relative to the total amount of the cosmetic product.

[0044] The skin bacterial flora-improving agent of the present invention described above is intended to be applied to mammals including humans (preferably humans). EXAMPLES

[0045] The present disclosure will be described in more detail below with reference to examples, although the present invention is not limited to these examples.

[0046] [Preparation of enzyme-treated royal jelly extract] Royal jelly was subjected to enzyme treatment according to the method described in WO2020 / 196065A1. Specifically, 400 mL of ion-exchanged water was added to 474 g of raw royal jelly and stirred until homogenous to prepare a royal jelly dilution. The pH of the royal jelly dilution was adjusted to 7.8 by adding a 2N NaOH aqueous solution. Next, a solution of 4.75 g of Proteax (Amano Enzyme Co., Ltd.), which has both endopeptidase and exopeptidase activities, dissolved in 20 mL of ion-exchanged water was added to the royal jelly dilution, and ion-exchanged water was further added so that the total amount was 980 g. The reaction mixture was reacted at 50 ° C (constant temperature water bath) for 2 hours while stirring with a propeller to perform hydrolysis. The temperature of the constant temperature water bath was raised to 80 ° C and stirred for 15 minutes to inactivate the protease. The obtained royal jelly enzyme-treated solution was filtered to a particle pore size of 3 μm using suction filtration or a filter press to obtain a royal jelly enzyme-treated filtrate. Butylene glycol, pentylene glycol, phenoxyethanol, and purified water were added to the royal jelly enzyme-treated filtrate so that the concentration of the royal jelly enzyme-treated filtrate was 2.5% by weight, and an enzyme-treated royal jelly extract was obtained.

[0047] [Preparation of test samples] A cosmetic containing enzyme-treated royal jelly extract (hereinafter referred to as RJ cream) and a placebo cosmetic (hereinafter referred to as placebo) were prepared with the formulations shown in Table 1. The units of values ​​are weight percent. 1,3-butylene glycol, pentylene glycol, and phenoxyethanol were contained in the enzyme-treated royal jelly extract in the same formulation ratio as the placebo.

[0048] [Table 1]

[0049] Forty-eight women aged 45 to 60 (average age 53.3 years) were asked to apply either RJ Cream or a placebo twice a day for 12 weeks to one half of their face, and the other test product was applied to the remaining half of their face under the same conditions. The effects of RJ Cream on the skin were examined by comparing the results with those of the placebo-applied side.

[0050] [Statistical analysis] Statistical analysis was performed for each evaluation item using the following method. The significance level was set at 5% using a two-sided test. For evaluation items with missing data, the data for all observation points for the relevant subjects were treated as missing values.

[0051] (intragroup comparison) For each evaluation item, the results before use of the test product were used as a control, and the results after use were tested using paired t-test or Wilcoxon signed-rank test. The significance level was adjusted according to the number of tests using the Bonferroni method.

[0052] (Intergroup comparison) For each evaluation item, the placebo-applied side of the same subject was used as a control, and the RJ-applied side was tested using Student's t-test.

[0053] [Test sample application] Approximately 1g of the test product was applied to half of the face twice a day (morning and evening) for 12 weeks. If participants normally used lotion or cream, they were asked to use the test product instead. The order of application of the skin care cosmetics and the method of application of the test product were as follows: (1) Daily use lotion (2) Essences, serums, etc. for everyday use (3) Test specimen Take half the amount (about 0.6g, 3 pumps) into your dry palms and gently and carefully apply it to half of your face, spreading it evenly. Next, take the other half (about 0.4g, 2 pumps) into your palms and gently and carefully apply it to half of your face, then gently and carefully apply it to the corners of your eyes, and finally wrap your hands together to blend it into your skin.

[0054] Skin bacterial flora samples were collected upon arrival at the testing site (before washing the face).

[0055] [Bacteria flora analysis] The resident bacteria on the cheek were collected using World Fusion's S-KIN kit (a swab soaked in saline) and subjected to S-KIN skin flora analysis. The species that make up the skin bacterial flora were identified by 16S rRNA metagenomic analysis. The calculated occupancy rate of each bacteria and the Shannon diversity index are shown in Table 2.

[0056] [Table 2]

[0057] It was confirmed that use of the test product containing enzyme-treated royal jelly extract reduced acne bacteria and Staphylococcus aureus in the skin bacterial flora, while increasing diversity.

Claims

1. A skin flora improver that contains enzyme-treated royal jelly as an active ingredient.

2. The skin bacterial flora improving agent according to claim 1, which is used to reduce the proportion of at least one species selected from the group consisting of Cutibacterium and Staphylococcus in the skin bacterial flora.

3. The skin bacterial flora improving agent according to claim 2 , wherein the Cutibacterium sp. bacterium is Propionibacterium acnes and the Staphylococcus sp. bacterium is Staphylococcus aureus.

4. The skin bacterial flora improving agent according to claim 1 or 2, which is used to improve the diversity of skin bacterial flora.

5. The skin bacterial flora improving agent according to claim 1 or 2, wherein the enzyme-treated royal jelly is treated with a peptidase having endopeptidase activity and exopeptidase activity.

6. The skin bacterial flora improving agent according to claim 1 or 2, which is for external use on the skin.

7. A cosmetic for improving skin flora, comprising the skin flora improving agent according to claim 1 or 2.

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