Agent for regulating skin-resident microbiota

Abstract

To provide a novel agent for regulating skin-resident microbiota, not being limited in use form to topical skin agents and enabling adjustment of the balance of skin resident microbiota in other forms.SOLUTION: Provided is an agent comprising essential oil and / or extract of coriander or bergamot.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to an agent for regulating resident skin bacteria, which is capable of preferentially regulating beneficial bacteria among resident skin bacteria. [Background technology]

[0002] The various bacteria present on human skin are called resident skin bacteria, and it has been suggested that the resident skin flora (microbiome) consisting of resident skin bacteria is involved in the health of the skin and skin-related diseases. Resident skin bacteria are broadly divided into beneficial bacteria represented by Staphylococcus epidermidis and harmful bacteria represented by Staphylococcus aureus.

[0003] For example, Patent Document 1 discloses a technology for selectively inhibiting the growth of harmful bacteria using a plant extract selected from among Peony root, Licorice, Oolong tea, Rosemary, Sophora flavescens, Coptis chinensis, Bark sprout, Thyme, Aloe, Chinese tallow tree, Mangosi, Cordyceps sinensis, Senso, Menyanthus trifoliata, Cleome, Trillium edulis, and Capsicum anguicum. Patent Document 2 discloses an ecosystem balance regulator for skin resident bacteria containing bittern as an active ingredient. It is described that the ecosystem balance regulator disclosed in Patent Document 2 can be further used in combination with rosemary extract or licorice extract. Patent Document 3 discloses an ecosystem balance regulator for skin resident bacteria containing Bitter melon or Pandanus plant extract and bittern as active ingredients. Patent Document 4 discloses an ecosystem balance regulator for skin resident bacteria containing hinokitiol glycoside as an active ingredient. Furthermore, Patent Document 5 discloses a glycan preparation suitable for local administration to sites other than the gastrointestinal tract, including mucosal tissue (oral cavity, nasal cavity, and vagina), and discloses that the glycan preparation regulates the abundance of bacterial taxa in the above-mentioned sites. Furthermore, Patent Document 6 discloses a skin flora balance improver that contains molasses or an extract thereof, and brewer's lees or an extract thereof, and is composed of a liquid with a sugar content of 60 to 85.

[0004] As described above, various biological substances have been developed to adjust the balance of skin flora. As disclosed in Patent Documents 1 to 6, the use of these substances has been limited to topical skin preparations containing various active ingredients. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] JP 2001-226213 A [Patent Document 2] JP 2005-139075 A [Patent Document 3] JP 2007-153800 A [Patent Document 4] JP 2007-246411 A [Patent Document 5] Special Publication No. 2018-532696 [Patent Document 6] Patent Publication No. 2021-195341 Summary of the Invention [Problem to be solved by the invention]

[0006] However, the form of use is not limited to skin topical preparations, and no active ingredient capable of adjusting the balance of skin resident bacteria in other forms has been known. In view of the above circumstances, the present invention aims to provide a novel skin resident bacteria regulator that is not limited to skin topical preparations, and can adjust the balance of skin resident bacteria in other forms. [Means for solving the problem]

[0007] In order to achieve the above-mentioned objective, the inventors conducted intensive research and succeeded in identifying plant-derived essential oils or extracts that have excellent effects as regulators of resident skin bacteria, thereby completing the present invention.

[0008] The present invention encompasses the following: (1) A skin flora regulator containing coriander or bergamot essential oil and / or extract. (2) A skin flora regulator according to (1), characterized in that the presence of Staphylococcus epidermidis is increased by the essential oil and / or the extract. (3) The skin resident flora regulator according to (1), wherein the essential oil and / or the extract has a growth-promoting effect on Staphylococcus epidermidis and / or a growth-inhibiting effect on Staphylococcus aureus. (4) A space having the effect of regulating skin flora, equipped with a device for sustained release of coriander or bergamot essential oil and / or extract. (5) A method for constructing a space having a regulating effect on resident skin bacteria by slowly releasing coriander or bergamot essential oil and / or extract into the space. Effect of the Invention

[0009] According to the present invention, it is possible to provide a regulator of resident skin bacteria that is not limited to a skin topical preparation and can be applied to various usage forms. By using the regulator of resident skin bacteria according to the present invention, it is possible to construct a space that has a regulating effect on resident skin bacteria. [Brief description of the drawings]

[0010] [Figure 1] FIG. 2 is a characteristic diagram showing the results of measuring lactate dehydrogenase (LDH) secreted into a medium when the plant-derived essential oil used in Example 1 was applied. [Diagram 2] FIG. 2 is a characteristic diagram showing the results of measuring the numbers of bacteria of Staphylococcus epidermidis and Staphylococcus aureus when the plant-derived essential oil used in Example 1 was applied. [Diagram 3] FIG. 2 is a characteristic diagram showing the results of measuring the inflammatory cytokine IL-1α secreted into the culture medium when the plant-derived essential oil used in Example 1 was applied. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0011] The skin flora regulator will be described in detail below. The regulator of skin flora includes essential oil and / or extract of coriander or bergamot. Essential oil and / or extract of coriander or bergamot has the effect of regulating skin flora. Here, essential oil means volatile oil produced from plants. Extract means including substances other than essential oil produced from plants. For example, the extract can be water-soluble substances extracted from plants using water or aqueous solutions containing said substances, and aqueous solutions (floral water) obtained when essential oil is produced by steam distillation.

[0012] The method for producing essential oils from plants is not particularly limited, and examples thereof include steam distillation, hot water distillation (direct distillation), squeezing, solvent extraction, and carbon dioxide extraction. Coriander and bergamot essential oils can be obtained by appropriately using these methods. Coriander and bergamot essential oils can also be obtained by mixing the oils obtained by a plurality of the methods listed above.

[0013] Coriander essential oil means an essential oil produced from coriander (Coriandrum sativum). Coriander essential oil can be produced from coriander seeds, leaves, flowers, stems, roots, or the whole plant, but is usually produced from seeds by steam distillation. The coriander variety used is not particularly limited, and any known variety of coriander can be used as appropriate.

[0014] Bergamot essential oil means an essential oil produced from bergamot (Citrus bergamia). Bergamot essential oil can be produced from bergamot peel, leaves, flowers, stems, roots, or the entire plant, but is usually produced from the peel by a pressing method or a steam distillation method. The type of bergamot used is not particularly limited, and any type of bergamot known in the art can be used as appropriate.

[0015] This skin flora regulator can regulate the balance of skin flora present on human skin. Here, the skin flora is composed of beneficial bacteria such as Staphylococcus epidermidis and harmful bacteria such as Staphylococcus aureus. The effect of regulating the skin flora by this skin flora regulator means that the proportion of beneficial bacteria in the total skin flora is increased. In other words, the effect of regulating the skin flora means one or both of the growth promotion effect on beneficial bacteria and the growth inhibition effect on harmful bacteria.

[0016] The method for measuring the effect of regulating the skin flora is not particularly limited, and any conventionally known method can be used as appropriate.For example, a three-dimensional cultured skin model in which Staphylococcus epidermidis and Staphylococcus aureus coexist can be used to measure the effect of regulating the skin flora of a given component.That is, the proportion of the skin flora present in the presence and absence of the component can be measured to measure the effect of regulating the skin flora of the component.

[0017] In addition, the effect of the component on regulating the resident skin bacteria can be measured by determining the skin toxicity of Staphylococcus aureus on the three-dimensional cultured skin model. In other words, if the skin toxicity is reduced in the presence of the component, it means that the proportion of Staphylococcus aureus is reduced, and it can be determined that the component has an effect on regulating the resident skin bacteria. The skin toxicity can be evaluated by colorimetric quantification of lactate dehydrogenase (LDH) released into the medium in which the three-dimensional cultured skin model is cultured.

[0018] Methods for measuring Staphylococcus epidermidis and Staphylococcus aureus that coexist in a three-dimensional cultured skin model include, for example, a measurement method using a flow cytometer or a method in which the number of colonies is counted after culturing on a solid medium.

[0019] The subject of use of the skin resident bacteria regulator containing essential oil and / or extract of coriander or bergamot is not particularly limited, and may be any mammal including human. By using this skin resident bacteria regulator, it is possible to obtain the effect of regulating the skin resident bacteria in any mammal including human. That is, by using this skin resident bacteria regulator, it is possible to increase the proportion of beneficial bacteria such as Staphylococcus epidermidis on the skin of any mammal including human. As a result, by using this skin resident bacteria regulator, it is possible to improve the health of the skin of any mammal including human. For example, by using this skin resident bacteria regulator, it is possible to expect effects such as suppressing the invasion of pathogenic bacteria into the skin, maintaining the barrier function of the skin, and protecting the skin from skin irritation. In addition, by using this skin resident bacteria regulator, it is possible to expect prevention and improvement of diseases related to skin resident bacteria, such as atopic dermatitis and acne vulgaris.

[0020] As described above, in order to enjoy various effects that can be expected by using the skin resident bacteria regulator, the skin resident bacteria regulator can be used in various forms. As an example, a use form in which a device for slowly releasing the skin resident bacteria regulator is arranged in a space is considered. That is, the space in which the device is arranged has the effect of the skin resident bacteria regulator described above. Here, the space means any space in which humans exist, such as a hospital room, a workplace, a commercial facility, and a private room, and any space in which mammals other than humans exist, such as a chicken farm, a cowshed, a pig farm, and a cage for pet animals. The space may also be a capsule-type space in which the device is arranged.

[0021] The device disposed in such a space is not particularly limited as long as it slowly releases a skin resident flora regulator, and may be a container with a liquid or gel skin resident flora regulator inside and a portion of the container open to the outside. The device may also be configured to include a woven or nonwoven fabric impregnated with a liquid skin resident flora regulator.

[0022] In addition, the skin flora regulator may be coriander or bergamot essential oil and / or extract itself, but may also be a composition containing a solvent such as ethanol, propylene glycol, glycerin, edible oils and fats, medium-chain fatty acids (MCTs), etc.

[0023] Furthermore, the skin flora regulator may be a composition containing various compounding agents and / or additives. The compounding agents and additives are not particularly limited, but examples thereof include antioxidants, known preservatives and antibacterial agents, pH adjusters, etc. The compounding agents and additives may be used in any combination of two or more types.

[0024] More specifically, antioxidants include butylhydroxytoluene, butylhydroxyanisole, citric acid, glutathione, selenium, lycopene, vitamin A, vitamin E, vitamin C, etc., as well as free radical scavengers obtained from pyrrolopyrrole derivatives and extracts from various plants, and enzymes with antioxidant properties, such as superoxide dismutase and glutathione peroxidase.

[0025] In addition, examples of preservatives and antibacterial agents include benzoic acid, sodium benzoate, isopropyl parahydroxybenzoate, isobutyl parahydroxybenzoate, ethyl parahydroxybenzoate, methyl parahydroxybenzoate, butyl parahydroxybenzoate, propyl parahydroxybenzoate, sodium sulfite, sodium hyposulfite, potassium metabisulfite, sorbic acid, potassium sorbate, sodium dehydroacetate, thujaplicin, Aralia udo extract, Styrax japonica extract, Artemisia capillaris extract, oolong tea extract, Shirako protein extract, enzymatically hydrolyzed Job's tears extract, and tea catechin. Examples of such antioxidants include apple polyphenols, pectin hydrolysates, chitosan, lysozyme, and ε-polylysine.

[0026] Examples of pH adjusters include adipic acid, citric acid, trisodium citrate, glucono-delta-lactone, gluconic acid, potassium gluconate, sodium gluconate, DL-tartaric acid, L-tartaric acid, potassium hydrogen DL-tartrate, potassium hydrogen L-tartrate, DL-sodium tartrate, sodium L-tartrate, potassium carbonate (anhydrous), sodium hydrogen carbonate, sodium carbonate, carbon dioxide, lactic acid, sodium lactate, glacial acetic acid, disodium dihydrogen pyrophosphate, fumaric acid, monosodium fumarate, DL-malic acid, sodium DL-malate, phosphoric acid, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, and sodium dihydrogen phosphate.

[0027] According to a usage form in which the device for slowly releasing the skin resident flora regulator is placed in a space, the space has the effect of regulating the skin resident flora, and the health condition of the skin in mammals, including humans, can be improved.

[0028] On the other hand, the present skin resident bacteria regulator can be provided as a medicine, cosmetic, or cosmetic product containing the above-mentioned composition. When the skin resident bacteria regulator is a medicine or cosmetic, it can be in the form of a skin topical composition. When it is a skin topical composition, examples of the skin topical composition include, in addition to medicines and cosmetics, body cleansers (e.g., solid cleansers such as solid soaps (solid soap compositions), body soaps, facial cleansers, hand soaps, shampoos, rinses, conditioners, and other liquid cleansers), hair creams, hair styling products, hair tonics, and the like.

[0029] The skin topical composition can be appropriately blended with various ingredients that can be commonly used in the fields of medicines, quasi-drugs, or cosmetics. When blending various ingredients, it is preferable to blend them within a quantitative and qualitative range that does not impair the effect of the skin flora regulator on the regulation of the skin flora. Examples of these ingredients include alcohols, ethers, esters, polyhydric alcohols, sugars, polymers, surfactants, spherical powders and coloring materials, plant and seaweed extracts, amino acids and peptides, vitamins, bactericides and preservatives, steroids, antioxidants, chelating agents, fragrances, water, etc. These ingredients can be blended in any combination of one or more kinds.

[0030] In addition, the skin external composition can contain other active ingredients in addition to the regulator of skin flora.The active ingredients are not particularly limited, but include, for example, nonsteroidal anti-inflammatory agents, whitening agents, anti-wrinkle agents, anti-inflammatory analgesics, antifungal agents, hair growth agents, slimming agents, local anesthetics, antipruritics, antibacterial agents, antiviral agents, keratin softeners, moisturizers, astringents, antioxidants, hair growth inhibitors, ultraviolet absorbing agents, ultraviolet scattering agents, etc.These ingredients can be used alone or in combination of two or more.

[0031] By using this skin resident flora regulator as a composition for external application to the skin, the health condition of the skin can be improved by regulating the skin resident flora in the subjects, mammals including humans. EXAMPLES

[0032] The present invention will be described in more detail below with reference to examples, but the technical scope of the present invention is not limited to the following examples.

[0033] Example 1 In this example, the effect of a certain plant-derived essential oil on skin flora was measured using a human three-dimensional cultured epidermis model. First, in this example, a skin flora symbiosis model was prepared in which Staphylococcus epidermidis and Staphylococcus aureus coexisted in a human three-dimensional cultured epidermis model (manufactured by Japan Tissue Engineering Co., Ltd., product name: LabCyte EPI-MODEL) as follows.

[0034] After replacing the medium with liquid medium, the LabCyte EPI-MODEL was incubated in CO for more than 15 hours. 2 The bacteria cultured in an incubator were used for evaluation. Various bacteria, S. aureus USA300 strain (S. aureus) and S. epidermidis ATCC35984 strain (S. epidermidis), were precultured in brain heart infusion medium and the OD 600 The culture was adjusted to 0.1 and incubated for 18 hours for evaluation. The culture was washed three times with saline and the S. aureus: 1.6 × 10 5 pieces / mL, S. epidermidis: 1.6×10 6 After adjusting the concentration with saline to 2 × 10 cells / mL, S. aureus: 2 × 10 3 pieces / well, S. epidermidis:2×10 4 The bacteria were inoculated into the three-dimensional epidermis model at OD 600 = 1 to 8 x 10 8 Calculated as cells / mL.

[0035] The effects of various plant-derived essential oils on skin flora were evaluated by measuring the skin damage caused by a skin flora symbiosis model. In this example, coriander essential oil (manufactured by Tree of Life, steam distillation), bergamot essential oil (manufactured by Tree of Life, pressing method), ginger essential oil (manufactured by Tree of Life, steam distillation), rosewood essential oil (manufactured by Tree of Life, steam distillation), geranium essential oil (manufactured by Tree of Life, steam distillation), and galbanum essential oil (manufactured by Tree of Life, steam distillation) were tested.

[0036] Specifically, the essential oils derived from the above plants were added to the culture medium of the skin resident flora symbiosis model at a concentration of 1% by weight, and lactate dehydrogenase (LDH) secreted into the culture medium was measured after 48 hours. LDH was measured using the Cytotoxicity LDH assay Kit-WST (manufactured by Dojindo Chemical Industries, Ltd.). 490 [nm] was measured using a microplate reader (TECAN, Spark).

[0037] The number of Staphylococcus epidermidis and Staphylococcus aureus bacteria on the skin resident flora symbiosis model was measured as follows. That is, the three-dimensional epidermis model after 48 hours of culture was washed with physiological saline, and the grown bacteria were collected. The collected liquid was subjected to various fluorescent staining using anti-Staphylococcus aureus (ab20920, Abcam), anti-gram-positive bacteria (ab15098, Abcam), goat anti-rabbit IgG antibody Alexa Fluor 488, and goat anti-rabbit IgG antibody Alexa Fluor 647, and then subjected to a flow cytometer (SONY, SA3800) to count the number of each bacteria.

[0038] The results of measuring LDH when various plant-derived essential oils were used are shown in Figure 1, and the results of measuring the number of Staphylococcus epidermidis and Staphylococcus aureus bacteria are shown in Figure 2. As shown in Figure 1, it was found that when coriander essential oil was used, the skin toxicity caused by Staphylococcus aureus was significantly reduced. Also, as shown in Figure 2, it was found that when coriander essential oil and bergamot essential oil were used, the proportion of Staphylococcus epidermidis, which is a beneficial bacteria among the skin resident bacteria, was significantly increased in the skin resident bacterial symbiosis model. In particular, when coriander essential oil and bergamot essential oil were used, the number of Staphylococcus aureus bacteria was significantly reduced, and as a result, skin toxicity was significantly reduced and the health of the skin was significantly improved.

[0039] Example 2 In this example, the effects of coriander essential oil and bergamot essential oil on skin inflammation caused by Staphylococcus aureus were evaluated using a human three-dimensional cultured epidermis model. First, in the same manner as in Example 1, a skin resident flora symbiosis model was prepared in which Staphylococcus epidermidis and Staphylococcus aureus coexisted in a human three-dimensional cultured epidermis model (manufactured by Japan Tissue Engineering Co., Ltd., product name: LabCyte EPI-MODEL).

[0040] Then, similarly to the examples, various plant-derived essential oils (coriander essential oil, bergamot essential oil, ginger essential oil, rosewood essential oil, geranium essential oil, and galbanum essential oil) were added to the medium at 1% by weight, and after culturing for 48 hours, the inflammatory cytokine IL-1α secreted into the medium was measured as follows: A human IL1-alpha measurement ELISA kit (Cosmo Bio, product number: KE00123) was used, and the absorbance at 450 nm was measured with a microplate reader (TECAN, Spark).

[0041] The results are shown in Figure 3. As shown in Figure 3, when coriander essential oil was used among the plant-derived essential oils tested, the concentration of the inflammatory cytokine IL-1α was significantly reduced. This result suggests that coriander essential oil can reduce the inflammatory effects of Staphylococcus aureus on the skin.

Claims

1. A skin flora regulator containing coriander or bergamot essential oil and / or extract.

2. The skin resident flora regulator according to claim 1, characterized in that the essential oil and / or the extract increases the presence rate of Staphylococcus epidermidis.

3. The skin resident flora regulator according to claim 1, characterized in that the essential oil and / or the extract has a growth-promoting effect on Staphylococcus epidermidis and / or a growth-inhibiting effect on Staphylococcus aureus.

4. A space having the effect of regulating the resident skin bacteria, and equipped with a device that slowly releases coriander or bergamot essential oil and / or extract.

5. A method for constructing a space having a regulating effect on resident skin bacteria by slowly releasing coriander or bergamot essential oil and / or extract into the space.

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