Balance regulator for skin microbiota

Abstract

To provide means of regulating or improving the balance of skin microbiota by applying in multiple manners a plant processed product such that it may exist in space.SOLUTION: One embodiment of the present invention relates to a balance regulator for skin microbiota, comprising a Salvia rosmarinus processed product as an active ingredient. Another embodiment of the present invention relates to a method for improving the skin condition of a subject, comprising applying the balance regulator for skin microbiota according to one embodiment of the present invention to the subject or to a space in which the subject exists.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a balance adjuster for resident skin bacteria. In particular, the present invention relates to a balance adjuster for resident skin bacteria containing processed rosemary as an active ingredient, and a method for improving the skin condition of a subject using the balance adjuster for resident skin bacteria. [Background technology]

[0002] Various bacteria exist on human skin. These bacteria are called resident skin flora. It has been suggested that the resident skin flora is related to skin homeostasis and / or disease. For example, in atopic dermatitis, the harmful bacterium Staphylococcus aureus (hereinafter also referred to as "S. aureus" or "S. aureus") is said to colonize the rash in approximately 90% of patients, and this bacterium has been linked to the worsening of the rash. On the other hand, Staphylococcus epidermidis (hereinafter also referred to as "S. epidermidis" or "S. epi") is known as a resident epidermal flora and is said to be a beneficial bacterium that enhances resistance to pathogenic bacteria by promoting the expression of antimicrobial peptides, etc. Therefore, it is important to increase or maintain the number of beneficial skin bacteria on human skin while simultaneously suppressing the number of harmful bacteria.

[0003] For example, Patent Document 1 describes a liquid agent for improving the balance of resident skin bacteria, which contains molasses or an extract thereof and alcohol brewer's lees or an extract thereof and has a sugar content of 60 to 85, and a liquid topical skin preparation containing the agent for improving the balance of resident skin bacteria. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Patent Publication No. 2021-195341 Summary of the Invention [Problem to be solved by the invention]

[0005] As described above, topical skin preparations containing specific active ingredients that adjust or improve the balance of resident skin bacteria are known. However, no means for adjusting or improving the balance of resident skin bacteria using processed plants that may be present in the air have been known. Furthermore, no means for adjusting or improving the balance of resident skin bacteria by other application methods, not limited to application to the skin, have been known.

[0006] Therefore, an object of the present invention is to provide a means for adjusting or improving the balance of normal skin flora by applying processed products of plants that may be present in the space in various ways. [Means for solving the problem]

[0007] The present inventors have investigated various means for solving the above problems. They have found that applying a processed rosemary product to a subject or the space in which the subject is present can increase the proportion of beneficial bacteria that are resident on the subject's skin and / or decrease the proportion of harmful bacteria, thereby adjusting the balance of resident skin bacteria. Based on these findings, the present inventors have completed the present invention.

[0008] That is, the present invention includes the following aspects and embodiments. (Embodiment 1) An agent for adjusting the balance of normal skin bacteria, comprising processed rosemary as an active ingredient. (Embodiment 2) The agent for adjusting the balance of resident skin bacteria according to embodiment 1, wherein adjusting the balance of resident skin bacteria includes at least one of increasing the proportion of Staphylococcus epidermidis and decreasing the proportion of Staphylococcus aureus among the resident skin bacteria. (Embodiment 3) The agent for adjusting the balance of resident skin bacteria according to embodiment 1 or 2, wherein the processed rosemary is a processed rosemary leaf. (Embodiment 4) The agent for adjusting the balance of resident skin bacteria according to any one of embodiments 1 to 3, wherein the processed product of rosemary is a water extract of rosemary leaves. (Embodiment 5) A method for improving the skin condition of a subject, comprising applying the skin resident flora balancing agent according to any one of embodiments 1 to 4 to the subject or a space in which the subject is present. [Effects of the Invention]

[0009] The present invention makes it possible to provide a means for adjusting or improving the balance of normal skin flora by applying processed products of plants that may be present in the space in various ways. [Brief explanation of the drawings]

[0010] [Figure 1] This figure shows the skin damaging activity of each sample against Staphylococcus aureus (S. aureus) in Example 1. In the figure, the horizontal axis represents the bacterial species and added samples on the human three-dimensional cultured epidermal model, and the vertical axis represents the skin damaging activity (LDH / OD490). [Figure 2] This figure shows the number of bacteria on a three-dimensional cultured human epidermal model to which each sample was added in Example 1. In the figure, the horizontal axis represents the bacterial species on the symbiotic model and the added sample, and the vertical axis represents the number of Staphylococcus epidermidis and Staphylococcus aureus bacteria (cell counts). In each bar, the upper row represents the number of Staphylococcus epidermidis (S. epi) bacteria, and the lower row represents the number of Staphylococcus aureus (S. aureus) bacteria. [Figure 3] This figure shows the number of bacteria on a three-dimensional cultured human epidermal model to which a water extract of rosemary leaves was added in Example 2. In the figure, the horizontal axis represents the bacterial species on the model and the dilution rate of the sample, and the vertical axis represents the number of bacteria (cell counts) of Staphylococcus epidermidis (S. epi) and Staphylococcus aureus (S. aureus). DETAILED DESCRIPTION OF THE INVENTION

[0011] Preferred embodiments of the present invention will now be described in detail.

[0012] <1. Skin flora balance adjuster> One aspect of the present invention relates to an agent for adjusting the balance of resident skin bacteria. The agent for adjusting the balance of resident skin bacteria of this aspect contains a processed product of rosemary (Salvia rosmarinus) as an active ingredient.

[0013] The subjects to which each aspect of the present invention is applied are preferably humans or non-human mammals (e.g., warm-blooded animals such as pigs, dogs, cows, rats, mice, guinea pigs, rabbits, chickens, sheep, cats, monkeys, hamadryas baboons, or chimpanzees), and more preferably humans.

[0014] In each aspect of the present invention, the term "resident skin bacteria" refers to a group of bacteria that are normally present on the skin of a subject. Resident skin bacteria can be classified into beneficial bacteria and harmful bacteria. Examples of beneficial bacteria that are resident skin bacteria include Staphylococcus epidermidis (S. epidermidis). Examples of harmful bacteria that are resident skin bacteria include Staphylococcus aureus (S. aureus).

[0015] In each aspect of the present invention, adjusting the balance of resident skin bacteria preferably includes improving the ratio of beneficial bacteria to harmful bacteria (beneficial bacteria / harmful bacteria ratio) among resident skin bacteria, more preferably improving the ratio of Staphylococcus epidermidis (S. epidermidis) to Staphylococcus aureus (S. aureus) (S. epidermidis / S. aureus ratio), and even more preferably at least one of increasing the proportion of S. epidermidis and decreasing the proportion of S. aureus. Resident skin bacteria on human skin have a defensive function to prevent the invasion of pathogenic bacteria from outside. On the surface of the skin, S. epidermidis has low pathogenicity and predominates over S. aureus, which is also a staphylococcus but is highly pathogenic. S. epidermidis produces fatty acids from sebum and sweat to maintain the epidermal surface in a slightly acidic state. This allows S. epidermidis to inhibit the growth of S. aureus and fungi (molds), which prefer a slightly alkaline environment. However, due to poor overall health, such as injury or illness, or some change in skin condition due to dryness or improper cleansing, the proper balance of skin flora may be disrupted. In such cases, excessive proliferation of resident bacteria and / or invasion or proliferation of other pathogenic bacteria may occur, potentially resulting in diseases or symptoms associated with resident skin bacteria, such as atopic dermatitis or acne vulgaris. Therefore, in each embodiment of the present invention, by adjusting the balance of resident skin bacteria, particularly Staphylococcus, in the subject exemplified above, it is possible to maintain the barrier function of the subject's skin, maintain the subject's skin in a healthy state, restore the subject's poor skin condition to a healthy state, and / or improve or prevent diseases or symptoms associated with resident skin bacteria, such as atopic dermatitis or acne vulgaris. This allows the skin condition of the subject to be improved by applying each embodiment of the present invention.

[0016] In each aspect of the present invention, the balance of resident skin bacteria can be evaluated, for example, by methods known in the art. Examples of such methods include an evaluation method using a three-dimensionally cultured human epidermal model seeded with either or both beneficial bacteria (e.g., Staphylococcus epidermidis) and harmful bacteria (e.g., Staphylococcus aureus). In an evaluation method using a three-dimensionally cultured human epidermal model, a sample is added to the model, the model is cultured for a certain period, and the number of bacteria is counted to evaluate the growth-promoting or growth-inhibiting effect of the sample on beneficial and harmful bacteria. Alternatively, the skin-damaging activity of the sample is evaluated by measuring a skin damage marker (e.g., lactate dehydrogenase (LDH)) secreted into the culture medium after culturing using the above procedure. The number of bacteria can be measured, for example, using a flow cytometer or a colony count method in which bacteria are seeded on a solid medium. The measurement of skin damage markers can be performed, for example, using a marker measurement kit commonly used in the art.

[0017] In each aspect of the present invention, the type, origin, place of origin, etc. of rosemary used as an active ingredient are not particularly limited. Rosemary of any type, origin, and place of origin can be used as an active ingredient.

[0018] In each embodiment of the present invention, the processed rosemary used as an active ingredient may be any part of the plant. The processed rosemary may be, for example, leaves, stems, flowers, or roots, particularly leaves. The parts of the rosemary plant exemplified above may be used alone or in combination of one or more.

[0019] In each embodiment of the present invention, the processed rosemary product used as the active ingredient may be an extract of the rosemary plant parts exemplified above, a separated or purified fraction thereof, or an active substance isolated therefrom. The extract of the rosemary plant parts is preferably an extract obtained by immersing the rosemary plant parts in water or an organic solvent such as alcohol, acetone, acetonitrile, diethyl ether, or ethyl acetate, or a volatile extract or essential oil obtained from the rosemary plant parts by steam distillation, hot water distillation (direct distillation), pressing, solvent extraction, or carbon dioxide extraction. An extract obtained by immersing the rosemary plant parts in pure water or an aqueous solution such as physiological saline (hereinafter also referred to simply as "aqueous extract") is more preferred, and an aqueous extract of rosemary leaves is even more preferred. The use of the processed rosemary products exemplified above as the active ingredient can adjust or improve the balance of resident skin bacteria.

[0020] The skin resident flora balancing agent of this embodiment may be in a form containing only processed rosemary as the active ingredient, or may be in the form of a composition containing processed rosemary and one or more additional ingredients.

[0021] The skin resident microbiota balancing agent of this embodiment can be provided in any form, such as a pharmaceutical, cosmetic, or fragrance. When the skin resident microbiota balancing agent of this embodiment is provided in the form of a pharmaceutical or cosmetic, it is preferably a composition for topical application to the skin. In the case of an embodiment of a composition for topical application to the skin, it is preferably in the form of a pharmaceutical or cosmetic preparation, or a body cleanser (for example, a solid cleanser such as a solid soap (solid soap composition), or a liquid cleanser such as a body soap, facial cleanser, hand soap, shampoo, rinse, or conditioner), hair cream, hair styling product, hair tonic, or the like.

[0022] In the case of an embodiment of a topical skin composition, the skin resident flora balance adjuster of this embodiment can optionally contain one or more additional ingredients commonly used in the fields of pharmaceuticals, quasi-drugs, or cosmetics. These one or more additional ingredients can include additives such as one or more of vehicles, salts or sugars, pH adjusters, buffers, antioxidants, preservatives, antibacterial agents, excipients, solubilizers, stabilizers, surfactants, emulsifiers, oily liquids (e.g., vegetable oils), suspending agents, chelating agents, sweeteners, and flavoring agents. When one or more of the above-mentioned additional ingredients are contained, it is preferable that the amount and quality of the ingredients be within a quantitative and qualitative range that does not impair the effect of the skin resident flora balance adjuster of this embodiment in regulating the skin resident flora.

[0023] For example, the medium may include water, alcohols such as ethanol, propylene glycol, glycerin, edible oils and fats, and medium-chain fatty acids.

[0024] For example, salts or sugars include sodium chloride, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, glucose, and the like.

[0025] Examples of pH adjusters include adipic acid, citric acid, trisodium citrate, gluconodeltalactone, gluconic acid, potassium gluconate, sodium gluconate, DL-tartaric acid, L-tartaric acid, potassium hydrogen DL-tartrate, potassium hydrogen L-tartrate, sodium DL-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.

[0026] Examples of antioxidants include butylhydroxytoluene, butylhydroxyanisole, citric acid, glutathione, selenium, lycopene, vitamin A, vitamin E, and vitamin C, as well as free radical scavengers obtained from pyrrolopyrrole derivatives or extracts of various plants, and enzymes with antioxidant properties such as superoxide dismutase and glutathione peroxidase.

[0027] Examples of preservatives or 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, Bleached syrup protein extract, enzymatically hydrolyzed Job's tears extract, tea catechins, apple polyphenols, pectin hydrolysates, chitosan, lysozyme, and ε-polylysine.

[0028] In the case of an embodiment of a topical skin composition, the skin resident flora balance adjuster of this aspect may contain one or more additional active ingredients in addition to the active ingredient, processed rosemary, such as nonsteroidal anti-inflammatory agents, whitening agents, anti-wrinkle agents, anti-inflammatory and analgesic agents, antifungal agents, hair growth agents, slimming agents, local anesthetics, antipruritics, antibacterial agents, antiviral agents, keratin softening agents, moisturizers, astringents, antioxidants, hair growth inhibitors, ultraviolet absorbers, and ultraviolet scattering agents.

[0029] The skin resident microbiota balancing agent of this embodiment is preferably applied to a subject or a space where the subject is present. When applying the skin resident microbiota balancing agent of this embodiment to a subject, it is preferable to apply or spray the skin resident microbiota balancing agent of this embodiment to the skin of the subject. In this case, the skin resident microbiota balancing agent of this embodiment is preferably an embodiment of a topical skin composition. Alternatively, the skin of the subject may be exposed to the skin resident microbiota balancing agent of this embodiment via an aroma bath or hot water bath containing the skin resident microbiota balancing agent of this embodiment. When applying the skin resident microbiota balancing agent of this embodiment to a space where a subject is present, it is preferable to spray the skin resident microbiota balancing agent of this embodiment into the gas phase present in the space and / or on the surface of an article. When spraying the skin resident microbiota balancing agent of this embodiment, it may be sprayed sequentially using a spray, or may be sprayed slowly and continuously using a nebulizer or diffuser.

[0030] Examples of spaces in which the subject exists include, for example, homes, hospital rooms, workplaces, and commercial facilities if the subject is a human; and poultry farms, cowsheds, pigpens, and pet cages if the subject is a non-human mammal.

[0031] <2. Method for improving the skin condition of a subject> Another aspect of the present invention relates to a method for improving the skin condition of a subject, which method comprises applying the agent for balancing resident skin bacteria of one aspect of the present invention to the subject or a space where the subject is present.

[0032] The method of this embodiment can be carried out by applying the skin resident flora balance adjuster of one embodiment of the present invention to a subject or a space in which the subject is present by the means exemplified above.

[0033] In the method of this aspect, by applying the agent for adjusting the balance of resident skin bacteria of one aspect of the present invention to a subject or a space in which the subject is present, the balance of resident skin bacteria of the subject can be adjusted to maintain the barrier function of the subject's skin, maintain the subject's skin in a healthy state, restore the subject's poor skin condition to a healthy state, and / or improve or prevent diseases or symptoms associated with resident skin bacteria, such as atopic dermatitis or acne vulgaris, thereby improving the subject's skin condition.

[0034] As described above in detail, by applying the agent for adjusting the balance of resident skin bacteria of one embodiment of the present invention to a subject or the space in which the subject is present, the balance of resident skin bacteria of the subject can be adjusted, thereby improving the subject's skin condition. Therefore, by implementing each embodiment of the present invention, a space (well-being space) that can improve the health of the subject can be created. [Example]

[0035] The present invention will be described in more detail below using examples, although the technical scope of the present invention is not limited to these examples.

[0036] [Example 1] A three-dimensional human epidermal model (product name: LabCyte EPI-MODEL12, manufactured by Japan Tissue Engineering Co., Ltd.) was used to create a symbiotic skin microbiota model by seeding the harmful bacterium Staphylococcus aureus (S. aureus) and the beneficial bacterium Staphylococcus epidermidis (S. epidermidis). Samples were added to this symbiotic skin microbiota model, and the skin-damaging activity of each sample derived from S. aureus was investigated. The samples used were processed rosemary, tea tree, and orange geranium. The processed rosemary, tea tree, or orange geranium samples were aqueous extracts obtained by extracting 3 g of rosemary, tea tree, or orange geranium leaves with 5 mL of saline at room temperature. Physiological saline was used as a control sample. Skin-damaging activity was assessed by adding each sample to a separate symbiotic model, culturing the model for 48 hours, and then measuring lactate dehydrogenase (LDH) secreted into the culture medium. LDH was measured using a commercially available kit that measures absorbance at 490 nm (OD 490). The skin-damaging activity of each sample derived from Staphylococcus aureus (S. aureus) is shown in Figure 1. In the figure, the horizontal axis represents the bacterial species and added sample in the symbiotic model, and the vertical axis represents the skin-damaging activity (LDH / OD 490).

[0037] As shown in FIG. 1, adding a water extract of rosemary leaves to the symbiotic model reduced the skin damaging activity compared to the control.

[0038] Bacteria were collected from the symbiotic model after 48 hours of culture. The number of collected Staphylococcus epidermidis (S. epi) and Staphylococcus aureus (S. aureus) bacteria was measured using a flow cytometer. Figure 2 shows the number of bacteria on the symbiotic model to which each sample was added. In the figure, the horizontal axis represents the bacterial species on the symbiotic model and the added sample, and the vertical axis represents the number of bacteria (cell counts) of Staphylococcus epidermidis and Staphylococcus aureus. In addition, the upper row of each bar represents the number of Staphylococcus epidermidis (S. epi) bacteria, and the lower row represents the number of Staphylococcus aureus (S. aureus) bacteria.

[0039] As shown in FIG. 2, it was confirmed that when the water extract of rosemary leaves was added to the symbiotic model, the S. epi / S. au ratio improved (beneficial bacteria dominance) compared to the control.

[0040] [Example 2] Using a human three-dimensional culture epidermal model similar to that in Example 1, we prepared separate models of resident skin bacteria by seeding the harmful bacterium Staphylococcus aureus or the beneficial bacterium Staphylococcus epidermidis. Samples were added to the S. aureus model or the S. epidermidis model to investigate the effect of each sample on cell proliferation. The same water extract of rosemary leaves as in Example 1 was diluted in several steps (×1, ×1 / 5, and ×1 / 20) and used as the added sample. After adding the samples to the separate models, the models were cultured for 48 hours. After 48 hours of culture, S. aureus or S. epidermidis were recovered from the models. The numbers of recovered S. epidermidis and S. aureus bacteria were counted using a flow cytometer. The numbers of bacteria on the models to which the water extract of rosemary leaves was added are shown in Figure 3. In the figure, the horizontal axis represents the bacterial species on the model and the dilution rate of the sample, and the vertical axis represents the bacterial count (cell count) of Staphylococcus epidermidis (S. epi) and Staphylococcus aureus (S. aureus).

[0041] As shown in FIG. 3, when a water extract of rosemary leaves was added to the model, a growth inhibitory effect was observed for Staphylococcus aureus and a slight growth promoting effect was observed for Staphylococcus epidermidis compared to the control.

[0042] The present invention is not limited to the above-described embodiments, but includes various modifications. For example, the above-described embodiments have been described in detail to clearly explain the present invention, and the present invention is not necessarily limited to those including all of the described configurations. Furthermore, it is possible to add, delete, and / or replace part of the configuration of each embodiment with other configurations.

Claims

1. A skin flora balancing agent containing processed rosemary as an active ingredient.

2. 2. The agent for adjusting the balance of resident skin bacteria according to claim 1, wherein the adjustment of the balance of resident skin bacteria includes at least one of increasing the proportion of Staphylococcus epidermidis and decreasing the proportion of Staphylococcus aureus among the resident skin bacteria.

3. 2. The agent for adjusting the balance of skin resident bacteria according to claim 1, wherein the processed rosemary is a processed rosemary leaf.

4. 2. The skin resident flora balance adjuster according to claim 1, wherein the processed rosemary is a water extract of rosemary leaves.

5. A method for improving the skin condition of a subject, comprising applying the skin resident flora balancing agent according to claim 1 to the subject or the space in which the subject is present.

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