Agent for improving permanent bacterial flora in skin and composition for external use on skin

Abstract

Provided is an agent for improving permanent skin microflora, which is useful for satisfactorily maintaining the state of the skin. The present invention is an agent for improving permanent skin flora using sophorose. The sophorose has the functions of inhibiting the growth of bacteria which may cause adverse effects on the skin in the resident bacteria of the skin and promoting the proliferation of beneficial bacteria which cause good effects on the skin in the resident bacteria of the skin. This agent for improving skin resident microflora can be suitably used for the purpose of maintaining the balance of skin resident microflora and maintaining the state of skin satisfactorily.

Description

Technical Field

[0001] The present invention relates to an agent for improving the resident flora of the skin and a composition for external use on the skin using sophora sugar. Background Art

[0002] It is known that various symbiotic microorganisms form groups and reside on the skin. Although there are individual differences, site differences, seasonal changes, etc., a certain balance is usually maintained, and the skin resident flora has an impact on skin function. The skin of a healthy person maintains the pH of the epidermis as weakly acidic, maintains a moisturizing environment, or improves resistance to pathogens through the skin resident flora, so that the skin condition is well maintained. On the other hand, if the balance of the skin resident flora is destroyed due to poor systemic conditions caused by trauma, disease, etc., mental stress, cold, dryness, improper washing, etc., not only will the skin's protective function against the outside world be reduced, but it may also induce various skin symptoms and cause disease manifestations.

[0003] In order to maintain the balance of the resident flora of the skin and maintain the skin function, the resident bacteria that undertake this task include, for example, Staphylococcus epidermidis, which is known as Staphylococcus epidermidis. This resident bacterium decomposes sebum, produces fatty acids that keep the skin weakly acidic, glycerol as a moisturizing component, or produces antimicrobial peptides that inhibit the proliferation of pathogenic bacteria, thereby improving the barrier function of the skin. Similarly, it is known that Staphylococcus hominis, which is known as Staphylococcus epidermidis, is also an epidermal resident bacterium that improves resistance to pathogenic bacteria by promoting the expression of antimicrobial peptides and the like.

[0004] On the other hand, among the bacteria resident on the skin, some bacteria are known to become harmful due to excessive proliferation. For example, Cutibacterium acnes, known as Propionibacterium acnes, has been reported to be useful for protecting the skin, but it is known that if the secretion of sebum increases or the pores are clogged, it will excessively proliferate and become the cause of acne vulgaris (pimples).

[0005] In addition, Staphylococcus aureus, also known as Staphylococcus aureus, is known to produce toxins that cause skin barrier damage and inflammation, and is present in large quantities in the skin of people with rough skin, people with dry skin, and patients with atopic dermatitis.

[0006] Corynebacterium xerosis, known as an actinomycete, is known to produce substances that cause malodor from sebum. Similarly, Micrococcus luteus, known as an actinomycete, is known to exist in large quantities on the skin of people with rough skin and to decompose sweat to produce malodorous substances.

[0007] As described above, the resident bacterial flora of the skin is considered to be extremely important in maintaining a good condition of the skin.

[0008] Regarding the resident flora of the skin, for example, Patent Document 1 records that oligosaccharides with an α1-2 sugar skeleton have the effect of promoting the proliferation of useful bacteria that reside on the skin and inhibiting the proliferation of undesirable bacteria such as odor-causing bacteria and pathogenic bacteria. In addition, Patent Document 2 discloses the invention of an agent for improving the resident flora of the skin containing oligomalto-oligosaccharides and / or sugar alcohols obtained by reducing oligomalto-oligosaccharides. In addition, Patent Document 3 discloses the invention of an agent for improving the resident flora of the skin containing oligoxyloses having uronic acid residues. In addition, Patent Document 4 records that cellobiose and a composition containing cello-oligosaccharides with cellobiose as the main component show antibacterial or proliferation-inhibiting effects on Staphylococcus aureus and Pseudomonas aeruginosa, but on the other hand, do not show antibacterial or proliferation-inhibiting effects on Staphylococcus epidermidis. In addition, Patent Document 5 discloses the invention of an agent for improving the balance of the resident flora of the skin with lactulose as an active ingredient.

[0009] Prior art literature

[0010] Patent Literature

[0011] Patent Document 1: Japanese Patent Application Publication No. 2002-113041

[0012] Patent Document 2: Japanese Patent Application Publication No. 2005-89355

[0013] Patent Document 3: Japanese Patent Application Publication No. 2007-77121

[0014] Patent Document 4: International Publication No. 2007 / 037249

[0015] Patent Document 5: International Publication No. 2021 / 132019 Summary of the invention

[0016] Problems to be solved by the invention

[0017] Conventionally, attempts to improve the flora of skin-resident bacteria using various oligosaccharides and the like have been known, but the development of new raw materials is desired for the convenience of consumers.

[0018] An object of the present invention is to provide an agent for improving the resident bacterial flora of the skin which is useful for maintaining a good condition of the skin.

[0019] Means for solving problems

[0020] The present inventors have conducted various studies to achieve the above-mentioned object, and as a result, have discovered sophora sugar as a raw material for selectively growing bacteria that have a good effect on the skin, thereby completing the present invention.

[0021] That is, the present invention, in its first aspect, provides an agent for improving the resident flora of the skin, comprising sophora sugar as an active ingredient.

[0022] In the present invention, the above-mentioned skin resident flora improving agent is preferably used to inhibit one or more species selected from Cutibacterium acnes, Staphylococcus aureus, Corynebacterium xerosis and Micrococcus luteus, and promote the proliferation of Staphylococcus epidermidis and / or Staphylococcus hominis.

[0023] On the other hand, the present invention provides, in its second aspect, a composition for external use on skin, comprising the above-mentioned agent for improving resident skin flora.

[0024] Effects of the Invention

[0025] According to the present invention, by using sophorose, the growth of bacteria that may have adverse effects on the skin among the skin-resident bacteria can be suppressed, and the proliferation of beneficial bacteria that have a good effect on the skin among the skin-resident bacteria can be promoted. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is a graph showing the results of investigating changes in bacterial counts when various bacteria that are resident bacteria on the skin were cultured under culture conditions suitable for each bacteria in Test Example 1 and α-glucooligosaccharide or β-glucooligosaccharide was added to the culture medium.

[0027] Figure 2 This is a graph showing the results of investigating changes in bacterial counts when Staphylococcus epidermidis, a skin resident bacterium, was cultured under culture conditions suitable therefor in Test Example 2 and highly purified sophorose, laminaribiose or gentiobiose was added to the culture medium.

[0028] Figure 3 This is a graph showing the results of investigating changes in bacterial counts when Staphylococcus epidermidis, a skin resident bacterium, was cultured under culture conditions suitable therefor in Test Example 3, and highly purified sophorose was added to the culture medium to give various concentrations of sophorose.

[0029] Figure 4 This is a graph showing the results of investigating changes in bacterial counts when various bacteria that are resident bacteria on the skin were cultured under culture conditions suitable for each bacteria in Test Example 4 and highly purified sophorose was added to the culture medium to give various concentrations of sophorose.

[0030] Figure 5 This is a graph showing the results of investigating changes in bacterial counts when various bacteria that are resident bacteria on the skin were cultured under culture conditions suitable for each bacteria in Test Example 5 and laminarin or gentiobiose prepared with high purity or commercially available cellobiose was added to the culture medium for culture. DETAILED DESCRIPTION

[0031] Hereinafter, the present invention will be described in more detail. It should be noted that in this specification, when there is a record of "~" about a numerical range, if there is no special description, it means any one of the numerical values ​​above to below and including both ends. In addition, the upper limit and the lower limit can be appropriately combined, and the numerical range thus obtained is also included in the disclosure of this specification.

[0032] The skin resident flora improving agent of the present invention uses sophorose as its active ingredient. Sophorose (English name: Sophorose, alias: 2-O-β-D-pyranose glucopyranosyl-D-glucose) is a disaccharide obtained by β-1,2 bonding of two glucose molecules. It was first discovered as a sugar component of the glycoside pigment of the pod of the leguminous plant Sophora japonica, but it rarely exists in the state of free disaccharides in nature and is a sugar that exists in very small amounts in the natural state. It is only known to exist rarely in acid hydrolyzates of starch and royal jelly.

[0033] There is no particular limitation on the acquisition of sophorose, and for example, it can be prepared using an enzyme by utilizing a glycosyl transfer / condensation reaction. Specifically, for example, as described in detail in Japanese Patent No. 2750374 and Japanese Patent No. 3020583, a β-glucosidase derived from a microorganism is allowed to act on glucose and / or β-glucooligosaccharide, and a condensation / glycosyl transfer reaction possessed by the β-glucosidase is utilized to obtain sophorose. In addition, as described in detail in Japanese Patent No. 7025941, by allowing 1,2-β-oligoglucan phosphorylase to act on α-glucose-1-phosphate and glucose, sophorose can be obtained at a higher yield.

[0034] By these methods, a composition containing sophorose is obtained. The obtained composition containing sophorose can be subjected to treatments such as filtration, decolorization, deodorization, and desalination as needed. In addition, the following treatments can also be carried out: extraction, centrifugation, crystallization, microbial assimilation, and classification by chromatography using activated carbon, porous carriers, hydrophobic resins, hydrophilic resins, ion exchange resins, adsorption resins, etc., dialysis, membrane classification such as ultrafiltration, etc. By these treatments, the purity of sophorose can be improved.

[0035] In the present invention, sophorose can also be used in the form of highly purified sophorose, but any substance containing sophorose can be used, that is, a sophorose-containing sugar composition can also be used. In this case, the content of sophorose in the sophorose-containing sugar composition used in the present invention is not particularly limited, for example, based on dry solid content, preferably 10% by mass or more, more preferably 30% by mass or more, further preferably 50% by mass or more, and most preferably 80% by mass or more. When the content of sophorose based on dry solid content is less than 5% by mass, the functionality of sophorose may not be fully exerted.

[0036] It should be noted that the sophorose-containing sugar composition used in the present invention may contain other components such as sugars and oligosaccharides having different bonding modes and polymerization degrees. Typically, for example, there can be exemplified a sugar composition containing one or more of sophorose oligosaccharides (Japanese: ソホロオリゴsugar) (excluding sophorose), gentiooligosaccharides, cellooligosaccharides, and laminarin oligosaccharides, etc., as oligosaccharides having a polymerization degree of 2 to 10, in the following dry solid content conversion content.

[0037] Sophora oligosaccharide: 5% to 80% by mass

[0038] Gentioligosaccharide: 5% to 50% by mass

[0039] Cellooligosaccharides: 5% to 50% by mass

[0040] Laminaria oligosaccharide: 5% to 50% by mass

[0041] As shown in the examples described below, sophorose has the functionality of inhibiting the growth of bacteria that may have adverse effects on the skin among the resident bacteria of the skin (also called "antibacterial"), and promoting the proliferation of beneficial bacteria that have a good effect on the skin among the resident bacteria of the skin. Specifically, there is no limitation on the beneficial bacteria that have a good effect on the skin, for example, Staphylococcus epidermidis, known as Staphylococcus epidermidis, Staphylococcus hominis, etc. On the other hand, as bacteria that may have adverse effects on the skin, there are Cutibacterium acnes, known as Propionibacterium acnes, Staphylococcus aureus, known as Staphylococcus aureus, Corynebacterium xerosis, known as actinomycetes, and Micrococcus luteus, known as actinomycetes.

[0042] In the present invention, since sophora sugar having the above-mentioned functionality is used as an active ingredient, it can be suitably used for the purpose of maintaining the balance of the resident bacterial flora of the skin and keeping the skin in good condition.

[0043] Specifically, in a certain mode, the present invention can be used for the purpose of improving the balance of the resident flora of the skin. If the balance of the resident flora of the skin is destroyed, it will cause excessive proliferation of the resident bacteria, cause the invasion and / or proliferation of other harmful bacteria, and induce various skin symptoms. However, according to the present invention, it can be used for the purpose of preventing from falling into such a state, or alleviating or suppressing the degree of the symptoms. In this case, the improvement of the balance can be carried out, for example, by increasing the proportion of Staphylococcus epidermidis, or reducing the proportion of Staphylococcus aureus. Staphylococcus epidermidis has an antagonistic effect on pathogenic bacteria, such as Staphylococcus aureus, and can prevent the proliferation of such harmful bacteria. Therefore, by promoting the proliferation of Staphylococcus epidermidis, it can play the role of a balance regulator of the resident flora of the skin. In addition, the improvement of the balance of the resident flora of the skin can also be, for example, not affecting the growth of beneficial bacteria such as Staphylococcus epidermidis, but inhibiting the growth of harmful bacteria such as Staphylococcus aureus.

[0044] In a certain embodiment, the present invention can be used to keep the skin weakly acidic, prevent infection by other bacteria, and prevent dryness. For example, by promoting the proliferation of Staphylococcus epidermidis, the bacteria decompose sebum, produce fatty acids that keep the skin weakly acidic, glycerin as a moisturizing component, or produce antimicrobial peptides that inhibit the proliferation of pathogenic bacteria, and thus can be used for the above purposes.

[0045] In one embodiment, the present invention can be used to suppress the production of toxins from harmful bacteria. For example, by suppressing the growth of Staphylococcus aureus, the bacteria can suppress the production of toxins, and thus can be used to suppress the above purpose.

[0046] In a certain embodiment, the present invention can be used for the purpose of preventing acne or alleviating or suppressing the degree of its symptoms. For example, by inhibiting the growth of Propionibacterium acnes, the onset of symptoms of acne vulgaris (acne) caused by the bacteria can be prevented, or the degree of symptoms can be alleviated or suppressed, and thus the present invention can be used for the above purpose.

[0047] In a certain embodiment, the present invention can be used for the purpose of preventing body odor, or reducing or suppressing its degree. For example, by suppressing the growth of Corynebacterium xerosis and Micrococcus luteus, which are known as actinomycetes, it is possible to suppress the bacteria from decomposing sebum and sweat to produce causative substances of bad odor, and thus it can be used for the above purpose.

[0048] In a certain aspect, the present invention is not limited to the prevention of specific skin symptoms, the alleviation and / or suppression of the degree of the symptoms, and can be used for the purpose of maintaining skin health, improving skin conditions, and imparting a barrier function to prevent pathogens from invading from the outside, for example.

[0049] (Composition for external use on skin)

[0050] The skin resident flora improving agent provided by the present invention can be used in the form of various skin external compositions such as cosmetics, external medicines, quasi-drugs, etc. Examples of skin external compositions include cosmetic creams, emulsions, lotions, masks, lotions, gels, powders, lipsticks, lip base products, foundations, sunscreen products, bath preparations, shampoos, conditioners, shower gels, body lotions, soaps, cleansing foams, ointments, patches, gels, aerosols, and the like.

[0051] When the skin resident flora improving agent provided by the present invention is used in the form of a skin external composition, the above-mentioned sophora sugar or sophora sugar-containing sugar composition can be prepared into respective forms according to conventional methods using appropriate pharmaceutically acceptable formulation carriers / ingredients. Examples of such formulation carriers / ingredients include: moisturizers such as glycerin, vaseline, urea, hyaluronic acid, and heparin; PABA derivatives (para-aminobenzoic acid, Escalol 507, etc.), cinnamic acid derivatives (Neo Heliopan, Parsol MCX, Sungard B, etc.), salicylic acid derivatives (octyl salicylate, etc.), benzophenone derivatives (ASL-24, ASL-24S, etc.), dibenzoylmethane derivatives (Parsol A, Parsol DAM, etc.), heterocyclic derivatives (Tinuvin series, etc.), titanium oxide and other ultraviolet absorbers / scatterers; metal chelators such as disodium edetate, trisodium edetate, citric acid, sodium citrate, tartaric acid, sodium tartrate, lactic acid, malic acid, sodium polyphosphate, sodium metaphosphate, gluconic acid, etc.; sebum inhibitors such as salicylic acid, sulfur, caffeine, tannin, etc.; bactericidal / disinfectants such as benzalkonium chloride, benzethonium chloride, chlorhexidine gluconate, etc.; anti-inflammatory agents such as diphenhydramine hydrochloride, tranexamic acid, guaiazulene, azulene, allantoin, cypress phenol, glycyrrhizic acid and its salts, glycyrrhizic acid derivatives, glycyrrhetinic acid, etc.; vitamins such as vitamin A, B vitamins (B1, B2, B6, B12, B15), folic acid, niacin, pantothenic acid, biotin, vitamin C, vitamin D vitamins (D2, D3), vitamin E, ubiquinones, vitamin K (K1, K2, K3, K4), etc. ; Amino acids and their derivatives such as aspartic acid, glutamic acid, alanine, lysine, glycine, glutamine, serine, cysteine, cystine, tyrosine, proline, arginine, pyrrolidone carboxylic acid, etc.; whitening agents such as retinol, tocopherol acetate, magnesium ascorbyl phosphate, ascorbyl glucoside, arbutin, kojic acid, ellagic acid, placenta extract, etc.; antioxidants such as butylated hydroxytoluene, butylated hydroxyanisole, propyl gallate, etc.; astringents such as zinc chloride, zinc sulfate, zinc carbolate, zinc oxide, potassium aluminum sulfate, etc.; sugars such as glucose, fructose, maltose, sucrose, trehalose, erythritol, mannitol, xylitol, lactitol, etc.; various plant extracts such as licorice, chamomile, horse chestnut, saxifrage, peony, papaya, scutellaria, phellodendron, coptis, houttuynia cordata, ginkgo leaf, etc.; as well as oily ingredients, surfactants, thickeners, alcohols, powder ingredients, pigments, etc. These formulation carriers / ingredients can be processed by conventional methods according to the type and form of the product to be added (for example, arbitrarily selecting and combining treatments such as pulverization, milling, washing, hydrolysis, fermentation, purification, pressing, extraction, classification, filtration, drying, powderization, granulation, dissolution, sterilization, pH adjustment, deodorization, and decolorization), and can be arbitrarily selected and supplied from a variety of raw materials.

[0052] It should be noted that when extracts (herbal medicines) from various plants or additives derived from animal raw materials are provided to the skin external composition of the present embodiment, in addition to the protection of the skin and hair, which can be expected to have moisturizing, improvement of touch / texture, provision of softness, relief of irritation, stress relief due to fragrance, cell activation (prevention of cell aging), suppression of inflammation, improvement of skin / hair quality, prevention and improvement of skin roughness, hair growth, hair growth, prevention of hair loss, provision of gloss, cleansing effect, relief of fatigue, promotion of blood flow, hot bath effect and other beauty effects, it is also possible to expect fragrance, deodorization, thickening, antiseptic, buffering and other effects.

[0053] Furthermore, in addition to these, various cosmetic effects and medicinal effects of the raw materials known so far can be expected, and by combining them, a product can be produced that is expected to have multifunctional effects in addition to the functionality of improving skin resident bacteria as the target of the present invention.

[0054] In the skin resident flora improving agent or skin external composition provided by the present invention, the content of sophorose can be in the range of 0.01% to 100% by mass based on dry solid content. When the content based on dry solid content is less than 0.01% by mass, the functionality based on sophorose may not be fully exerted. As the content of sophorose, it is more preferably 0.6% by mass or more, and further more preferably 0.75% by mass or more.

[0055] When the skin resident flora improving agent or skin external composition provided by the present invention is in the form of liquid, gel, cream or other medium containing water, the content of sophora sugar in such a form can be appropriately set without particular limitation, and typically, for example, can be in the range of 0.01 mg / g to 400 mg / g (the denominator represents the weight of the preparation), can be in the range of 0.1 mg / g to 100 mg / g, or can be in the range of 1 mg / g to 50 mg / g.

[0056] When the skin resident flora improving agent or skin external composition provided by the present invention is applied to the skin, the dosage thereof can be appropriately set without particular limitation. For example, it can be 0.001 mg / cm 2 ~1mg / cm 2 (the denominator represents the area of ​​the skin to which it is applied) can range from 0.005 mg / cm 2 ~0.5mg / cm 2 The range can be 0.02mg / cm 2 ~0.05mg / cm 2 range.

[0057] Example

[0058] The following are examples for a more specific description of the present invention, but these examples do not limit the scope of the present invention.

[0059] 〔1. Preparation of α-gluco-oligosaccharide〕

[0060] To a 75 (w / w)% glucose solution, 50 mg of glucoamylase preparation was added per 1 g of solid component. After adjusting the pH to 5.0, it was maintained at 66 °C. After reacting for 63 hours, the pH was adjusted to 4.0 and the temperature was raised to 80 °C to inactivate the enzyme. Monosaccharides were removed from the product by membrane fractionation to obtain α-gluco-oligosaccharide.

[0061] 〔2. Preparation of β-gluco-oligosaccharide〕

[0062] To a 60 (w / w)% glucose solution, 9.3 mg of β-glucosidase preparation was added per 1 g of solid component. After adjusting the pH to 6.6, it was maintained at 60 °C. After reacting for 72 hours, the pH was adjusted to 4.0 and the temperature was raised to 80 °C to inactivate the enzyme. Monosaccharides were removed from the product by membrane fractionation to obtain β-gluco-oligosaccharide.

[0063] 〔3. Preparation of high-purity sophorose〕

[0064] 1.0 M sucrose and 1.0 M glucose were dissolved in 100 mM potassium phosphate buffer (pH 7.0). To each 1 mL of this substrate, 10 μL of a protein having 1,2-β-oligoglucan phosphorylase activity derived from Bifidobacterium scardobii (the protein disclosed as "BSGP" in Japanese Patent No. 7025941) at 19 mg / mL and 10 μL of sucrose phosphorylase (Sigma-Aldrich) at 17 mg / mL were added, and the reaction was carried out at 30 °C. After 7 days, the enzyme was inactivated by boiling for 10 minutes, the reaction solution was appropriately concentrated, and it was applied to an activated carbon column to remove monosaccharides and disaccharides. Hydrochloric acid was added thereto to a final concentration of 85 mM, and it was maintained at 80 °C for 17 hours for acid decomposition. The decomposed sample was applied to HPLC under the following conditions, and the disaccharide fraction was recovered to obtain high-purity sophorose.

[0065] <HPLC conditions>

[0066] Column: ULTRON PS-80NL (Showa Chemical Industry Co., Ltd.)

[0067] Column temperature: 50 °C

[0068] Eluent: ultrapure water

[0069] Flow rate: 0.9 mL / min

[0070] Detector: differential refractive index detector

[0071] Analysis time: 20 minutes

[0072] [4. Preparation of High-Purity Laminaribiose]

[0073] Mix 8.0 g of curdlan (Mitsubishi Corporation Life Science Co., Ltd.) with 720 mL of ultrapure water, add 8 mL of 1 M sodium hydroxide, and stir for 10 minutes. After adding 40 mL of 1 M Tris-HCl buffer (pH 7.0) and 8 mL of 1 M hydrochloric acid and stirring, add 80 mg of yeast cell wall lytic enzyme (“Zymolyase (registered trademark)”, Nacalai Tesque, Inc.). Add cysteine to a final concentration of 10 mM and react overnight at 37°C with vigorous stirring. Heat at 80°C for 10 minutes to inactivate the enzyme, cool to room temperature, and then centrifuge to recover the supernatant. Subject it to HPLC under the same conditions as for the preparation of high-purity sophorose, recover the disaccharide fraction, and obtain high-purity laminaribiose.

[0074] [5. Preparation of High-Purity Gentiobiose]

[0075] Use the β-gluco-oligosaccharide mainly composed of disaccharide prepared in [2. Preparation of β-Gluco-Oligosaccharide] above as a raw material, perform fractionation using an activated carbon column, and obtain high-purity gentiobiose.

[0076] [6. Sugar Composition Analysis]

[0077] Sugar composition analysis is performed using HPLC. The content of each degree of polymerization component is determined from the peak area of the obtained chromatogram. The analysis of gluco-oligosaccharide is carried out according to HPLC condition A, and the analysis of high-purity samples is carried out according to HPLC condition A and HPLC condition B.

[0078] [HPLC Condition A]

[0079] Column: ULTRON PS-80NL (Showa Denko K.K.)

[0080] Column temperature: 50°C

[0081] Eluent: Ultrapure water

[0082] Flow rate: 0.9 mL / min

[0083] Detector: Differential refractive index detector

[0084] Analysis time: 20 minutes

[0085] [HPLC Condition B]

[0086] Column: HILICpak VG-50 4E (Showa Denko K.K.)

[0087] Column temperature: 40°C

[0088] Eluent: acetonitrile: ultrapure water = 8:2

[0089] Flow rate: 0.6mL / min

[0090] Detector: Charged particle detector

[0091] Analysis time: 60 minutes

[0092] Tables 1 and 2 show the results of sugar composition analysis.

[0093]

[0094] As shown in Table 1, the oligosaccharides prepared above all have a sugar composition with disaccharides as the main component.

[0095]

[0096] As shown in Table 2, the high-purity preparations of sophorose, laminaribiose and gentiobiose prepared above all had a purity of at least 90% by mass.

[0097] [Test Example 1]

[0098] When various bacteria shown in Table 3 as skin resident bacteria were cultured under culture conditions suitable for each bacteria, changes in bacterial counts were investigated when the α-glucooligosaccharide or β-glucooligosaccharide prepared above was added to the culture medium.

[0099]

[0100] Specifically, various bacteria were cultured overnight in a medium suitable for each bacterium to prepare a pre-culture solution, and α-glucooligosaccharide or β-glucooligosaccharide prepared above was added to 3 mL of fresh medium to a final concentration of 0.5 (w / v)%, and 0.03 mL of the pre-culture solution was further mixed therein, and cultured for 24 hours, and the bacterial turbidity (OD660) after culture was measured. In addition, glucose was added to the medium as a sugar and the test was performed in the same manner as a comparative control. The results are expressed as relative values ​​(relative to the sugar-free area %) when the bacterial turbidity when sterilized ultrapure water is added instead of sugar is set to 100%.

[0101] As a result, if Figure 1As shown in the figure, in the glucose addition area, S. epidermidis and S. hominis proliferated significantly, and the growth of S. aureus, C. acnes, and C. xerosis was also confirmed, which is the result of lack of selectivity for specific bacterial species. In addition, in the α-glucooligosaccharide addition area, in addition to S. epidermidis, the growth of S. aureus and C. xerosis was also confirmed. On the other hand, in the β-glucooligosaccharide addition area, the growth of S. epidermidis and S. hominis was confirmed, but the growth of the other four bacterial species was almost not confirmed.

[0102] As described above, it is clear that β-glucoligosaccharide, which is mainly composed of disaccharides, has the effect of inhibiting the growth of bacteria that are harmful to the skin and promoting the proliferation of Staphylococcus epidermidis (S. epidermidis, S. hominis) that are beneficial to the skin.

[0103] [Test Example 2]

[0104] The effect of promoting the proliferation of S. epidermidis was evaluated for sophorose, laminaribiose, and gentiobiose, which are β-glucooligosaccharides formed by disaccharides. Specifically, as the sugar added to the culture medium, sophorose, laminaribiose, or gentiobiose prepared with high purity as described above was used, and the amount added to the culture medium was set to a final concentration of 0.3 (w / v)% and 0.5 (w / v)%, and the same operation as in Experimental Example 1 was performed to evaluate the effect of promoting the proliferation of S. epidermidis.

[0105] As a result, if Figure 2 As shown, among various β-glucoligosaccharides (disaccharides), sophorose has a significant effect on promoting the proliferation of S. epidermidis. On the other hand, laminaribiose and gentiobiose have no effect on promoting the proliferation of S. epidermidis.

[0106] [Test Example 3]

[0107] The effect of promoting the proliferation of S. epidermidis was evaluated in the same manner as in Test Example 2 except that the sophorose prepared with high purity as described above was used as the disaccharide β-glucopyranosaccharide and the amount added to the medium was adjusted to a final concentration of 0.1 (w / v) % or 1.0 (w / v) %.

[0108] As a result, if Figure 3As shown in the example, when a preparation containing sophorose at a high purity was added to a medium to a final concentration of 0.1 (w / v)% and cultured, an effect of promoting the proliferation of S. epidermidis was also confirmed. On the other hand, when the final concentration was 1.0 (w / v)%, no significant difference was confirmed compared to the effect of promoting the proliferation of S. epidermidis shown in Test Example 2 when the final concentration was 0.5 (w / v)%.

[0109] [Test Example 4]

[0110] The effect of promoting the proliferation of S. hominis, S. aureus, C. acnes, C. xerosis or M. luteus was evaluated in the same manner as in Test Example 1, except that the sophorose prepared with high purity as described above was used as the β-glucopyranose formed from a disaccharide and the amount added to the medium was adjusted to a final concentration of 0.1 (w / v)%, 0.5 (w / v)% or 1.0 (w / v)%.

[0111] As a result, if Figure 4 As shown, by adding a preparation containing highly purified sophorose to the medium at a final concentration of 0.5 (w / v)% or 1.0 (w / v)%, the proliferation of S. hominis increased to 118.0% and 118.8%, respectively, as expressed as a relative value to the control to which no sugar was added (% relative to the sugar-free zone). On the other hand, no effect of promoting the proliferation of S. aureus, C. acnes, C. xerosis, or M. luteus was observed at any addition amount compared to the control.

[0112] As described above, when the results of Test Examples 2 and 3 are considered together, it is clear that sophorose, which is a disaccharide β-glucopyranosaccharide, has the effect of inhibiting the growth of bacteria harmful to the skin and promoting the proliferation of Staphylococcus epidermidis (S. epidermidis, S. hominis) beneficial to the skin.

[0113] [Test Example 5]

[0114] As β-glucopyranose formed from disaccharides, the above-prepared high-purity laminarin and gentiobiose and commercially available cellobiose (Nacalai Tesque Co., Ltd.) were used, and the addition amount to the medium was adjusted to a final concentration of 0.5 (w / v)%, and the effect of promoting the proliferation of S. hominis, S. aureus, C. acnes, C. xerosis, or M. luteus was evaluated in the same manner as in Test Example 1.

[0115] As a result, if Figure 5 As shown in the results, no proliferation-promoting effect was observed with the addition of laminarin, gentiobiose, and cellobiose. In addition, in the laminarin addition area, the proliferation of S. aureus increased to 108.5% as a relative value relative to the control without sugar addition (% relative to the sugar-free area), and in the gentiobiose and cellobiose addition areas, the proliferation of M. luteus increased to 121.9% and 111.7% as a relative value relative to the sugar-free control (% relative to the sugar-free area), respectively.

[0116] From the above, when the results of Test Example 2 are considered together, it can be seen that the effects of laminaribiose, gentiobiose, and cellobiose, which are β-glucooligosaccharides formed from disaccharides, in inhibiting the growth of bacteria harmful to the skin and promoting the proliferation of Staphylococcus epidermidis (S. epidermidis, S. hominis) beneficial to the skin are inferior to sophorose.

[0117] (Formulation Example 1)

[0118] This is a formulation example of using sophora sugar in a lotion as the agent for improving the resident skin flora of the present invention.

[0119] (Raw material name) Content (mass %)

[0120] Sophora sugar 0.5%

[0121] Betaine 1.0%

[0122] Glycerin 5.0%

[0123] 1,3-Butanediol 10.0%

[0124] Sodium carboxymethyl dextrin 0.1%

[0125] Dipotassium glycyrrhizinate 0.1%

[0126] Purified water balance

[0127] (Formulation Example 2)

[0128] This is a formulation example of using sophora sugar in shampoo as the skin resident flora improving agent of the present invention.

[0129] (Raw material name) Content (mass %)

[0130] Sophora sugar 0.5%

[0131] Betaine 0.5%

[0132] Sodium lauroyl aspartate solution 45.0%

[0133] Sodium lauroyl alanine 12.5%

[0134] Polyoxyethylene hexadecyl octadecyl diether 1.0%

[0135] Laureth-1,5 Glyceryl 1.0%

[0136] Citric acid 0.5%

[0137] Polyquaternium-10 0.8%

[0138] Methylparaben 0.2%

[0139] EDTA-2Na 0.1%

[0140] Purified water balance

[0141] (Formulation Example 3)

[0142] This is a formulation example of an agent for improving the resident skin flora of the present invention, in which sophora sugar is used in an emulsion.

[0143] (Raw material name) Content (mass %)

[0144] Sophora flavescens 1.0%

[0145] Betaine 1.0%

[0146] Glycerin 8.0%

[0147] 1,3-Butanediol 5.0%

[0148] Squalane 3.0%

[0149] Olive oil 1.0%

[0150] Cetyl Caprylate 5.0%

[0151] Stearic acid 0.3%

[0152] Polydimethylsiloxane 0.5%

[0153] Polysorbate 60 0.5%

[0154] Glyceryl monostearate 1.0%

[0155] Carboxyvinyl polymer 0.1%

[0156] Potassium hydroxide 0.01%

[0157] Purified water balance

Claims

1. A skin resident flora improving agent, which contains sophora sugar as an active ingredient.

2. The agent for improving the resident skin flora according to claim 1, which is used to inhibit one or more selected from Cutibacterium acnes, Staphylococcus aureus, Corynebacterium xerosis and Micrococcus luteus, and promote the proliferation of Staphylococcus epidermidis and / or Staphylococcus hominis.

3. A composition for external use on the skin, comprising the agent for improving the resident skin flora according to claim 1 or 2.

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