Vaginal microbiome balance improving agent and vaginal composition

Abstract

This invention provides an agent that improves the balance of the vaginal microbiome, which is useful for maintaining a healthy vaginal condition. [Solution] This is a vaginal microbiota balance improving agent using sophorose and / or laminaribiose. Sophorose and / or laminaribiose have the functionality to suppress the growth of bacteria in the vaginal microbiota that may have a negative impact on the vagina, while promoting the growth of beneficial bacteria that have a positive impact on the vagina. This vaginal microbiota balance improving agent can be suitably used to maintain the balance of the vaginal microbiota and keep the vagina in good condition.

Description

Technical Field

[0004] , , , , ,

[0003]

[0001] The present invention relates to a vaginal resident flora balance improver and a vaginal composition using sophorose and / or laminaribiose.

Background Art

[0002] In the bacterial flora of the vagina of a healthy human, many are occupied by bacteria of the genus Lactobacillus, and the vagina is kept acidic by lactic acid produced by the bacteria of the genus Lactobacillus. When the balance of the bacterial flora in the uterus is disrupted, bacterial vaginosis (BV) develops, causing itching and inflammation. Bacterial vaginosis (BV) can cause miscarriage and premature birth, and the infection can spread to other organs and become severe in some cases. Bacterial vaginosis (BV) is said to occur in one out of three women, and it is a disease with a high recurrence rate even after treatment with antibiotics.

[0003] Examples of bacteria of the genus Lactobacillus that form the bacterial flora of the human vagina include Lactobacillus gasseri, Lactobacillus paragasseri, Lactobacillus crispatus, and Lactobacillus jensenii. These bacteria of the genus Lactobacillus produce D-lactic acid and bacteriocin, maintaining the health of the vagina.

[0004] On the other hand, some of the normal vaginal flora are known to become harmful when they proliferate excessively. For example, Lactobacillus iners produces L-lactic acid, but it is insufficient to inhibit bacterial vaginosis (BV) and is known to easily lead to BV. Gardnerella vaginalis has been observed to proliferate in patients with bacterial vaginosis (BV) and is known to form biofilms that inhibit BV treatment. Mobiluncus curtisii has been observed to proliferate in patients with bacterial vaginosis (BV) and is known to be resistant to metronidazole, a treatment for bacterial vaginosis (BV). In addition, Atopobium vaginae has also been observed to proliferate in patients with bacterial vaginosis (BV).

[0005] For the reasons described above, the vaginal microbiota is considered to be extremely important in maintaining good vaginal health.

[0006] Regarding the vaginal flora, for example, Patent Document 1 discloses an invention for maintaining the pH value of vaginal secretions within the range of 3.5 to 4.5 by using a combination of sugars and at least one of benzoic acid and its sodium salt. Patent Document 2 discloses an invention for a formulation for preventing and / or treating infectious diseases of the vagina, containing a bacteriostatic agent including phenethyl alcohol as an active ingredient. Patent Document 3 discloses an invention for a lactic acid bacteria strain for the prevention and / or treatment of vaginal infections. [Prior art documents] [Patent Documents]

[0007] [Patent Document 1] Special Publication No. 2008-539176 [Patent Document 2] Special Publication No. 2019-534896 [Patent Document 3] Special Publication No. 2024-503366 [Overview of the project] [Problems that the invention aims to solve]

[0008] While attempts to improve the vaginal flora using bacteriostatic agents and lactic acid bacteria strains have been known, the development of new materials is desired for the convenience of consumers.

[0009] The object of the present invention is to provide an agent for improving the balance of the vaginal microbiota, which is useful for maintaining a good vaginal condition. [Means for solving the problem]

[0010] To achieve the above objective, the inventors conducted various studies and, as a result, discovered sophorose and / or laminaribiose as materials that selectively promote the growth of bacteria that have a beneficial effect on the vagina, thus completing the present invention.

[0011] In other words, in its first aspect, the present invention provides a vaginal microbiome balance improving agent comprising sophorose and / or laminaribiose as active ingredients.

[0012] In the present invention, it is preferable that the vaginal flora balance improving agent is used to promote the growth of one or more species selected from the group consisting of Lactobacillus gasseri, Lactobacillus paragasseri, Lactobacillus crispatus, and Lactobacillus jensenii, while stabilizing one or more species selected from the group consisting of Lactobacillus iners, Gardnerella vaginalis, Mobiluncus curtisii, and Atopobium vaginae.

[0013] Furthermore, it is preferable that the sophorose and / or laminaribiose used as the active ingredient is in the form of a composition containing sophorose and / or laminaribiose produced using a glycosyltransferase / condensation enzyme.

[0014] On the other hand, in a second aspect thereof, the present invention provides a vaginal composition containing the above-mentioned vaginal microbiome balance improving agent. [Effects of the Invention]

[0015] According to the present invention, by using sophorose and / or laminaribiose, it is possible to suppress the growth of bacteria that may have a negative impact on the vagina among the normal vaginal flora, while promoting the growth of beneficial bacteria that have a positive impact on the vagina. [Brief explanation of the drawing]

[0016] [Figure 1] This figure shows the results of an investigation into the changes in bacterial counts when various bacteria, considered as commensal vaginal flora, were cultured under appropriate culture conditions, with α-gluco-oligosaccharide or β-gluco-oligosaccharide added to the culture medium, as in Test Example 1. [Figure 2] This figure shows the results of an investigation into the changes in bacterial counts when various bacteria (L. crispatus, L. paragasseri, L. jensenii, L. gasseri) were cultured under appropriate culture conditions for each species, with high-purity sophorose or laminaribiose added to the culture medium, in Test Example 2. [Figure 3] This figure shows the results of an investigation into the changes in bacterial counts when various bacteria (L. crispatus, L. paragasseri, L. jensenii, L. gasseri) were cultured under appropriate culture conditions for each species, and high-purity sophorose or laminaribiose was added to the culture medium at varying final concentrations during the cultivation process in Test Example 3. [Figure 4]In Test Example 4, when culturing various bacteria (Lactobacillus iners, Gardnerella vaginalis, Mobiluncus curtisii, Atopobium vaginae) as vaginal resident bacteria under culture conditions suitable for each, the chart shows the results of examining the change in the number of bacteria when sophorose or laminaribiose prepared with high purity was added to the medium at different final concentrations and cultured.

Mode for Carrying Out the Invention

[0017] Hereinafter, the present invention will be described in more detail. In this specification, when there is a description of "~" for a numerical range, unless otherwise specified, it represents from the upper limit to the lower limit, including both end values. Also, the upper limit value and the lower limit value can be appropriately combined, and the numerical range obtained thereby is also included in the disclosure of this specification.

[0018] The vaginal resident flora balance improver according to the present invention uses sophorose and / or laminaribiose as its active ingredient.

[0019] Sophorose (English name: Sophorose, alias: 2-O-β-D-glucopyranosyl-D-glucose) is a disaccharide in which two molecules of glucose are β-1,2-linked. It was first found as the sugar component of the glycoside of the pigment in the sheath of the leguminous plant vetch, but it rarely exists in the free disaccharide state in nature and is a sugar that exists only very rarely naturally. It is only known to exist slightly in the acid hydrolysis product of starch or in royal jelly.

[0020] The acquisition of sophorose is not particularly limited. For example, it can be enzymatically prepared by utilizing glycosyl transfer and condensation reactions. Specifically, as described in detail in, for example, Japanese Patent No. 2750374 and Japanese Patent No. 3020583, sophorose can be obtained by allowing a β-glucosidase of microbial origin to act on glucose and / or β-glucooligosaccharide and utilizing the condensation and glycosyl transfer reactions possessed by the β-glucosidase. Further, as described in detail in Japanese Patent No. 7025941, according to the method of allowing 1,2-β-oligoglucan phosphorylase to act on α-glucose-1-phosphate and glucose, sophorose can be obtained in a higher yield.

[0021] Laminaribiose (English name: Laminaribiose, alias: 3-O-β-D-glucopyranosyl-D-glucose) is a disaccharide in which two molecules of glucose are β-1,3-linked, and is known as the lowest structural unit of β-1,3 glucans such as laminaran, curdlan, and pachyman.

[0022] The acquisition of laminaribiose is not particularly limited. For example, it can be enzymatically prepared by utilizing glycosyl transfer and condensation reactions. Specifically, as described in detail in, for example, Japanese Patent No. 2750374 and Japanese Patent No. 3020583, laminaribiose can be obtained by allowing a β-glucosidase of microbial origin to act on glucose and / or β-glucooligosaccharide and utilizing the condensation and glycosyl transfer reactions possessed by the β-glucosidase. Further, as described in detail in Japanese Patent No. 2955590, by using sucrose as a raw material and allowing three or four enzymes, namely sucrose phosphorylase, glucose isomerase, laminaribiose phosphorylase, and / or 1,3-β-oligoglucan phosphorylase, to act in combination in the presence of phosphoric acid, laminaribiose can be easily obtained in a high yield.

[0023] Compositions containing sophorose and / or laminaribiose can be obtained by these methods. The obtained compositions containing sophorose and / or laminaribiose can be subjected to treatments such as filtration, decolorization, deodorization, and desalting as needed. They may also be subjected to treatments such as extraction, centrifugation, crystallization, microbial assimilation, and further treatments such as chromatographic fractionation using activated carbon, porous carriers, hydrophobic resins, hydrophilic resins, ion exchange resins, adsorption resins, etc., dialysis, and membrane fractionation using extreme filtration. These treatments can increase the purity of sophorose and / or laminaribiose.

[0024] In the present invention, sophorose and / or laminaribiose can be used in a highly purified form, but any product containing sophorose and / or laminaribiose is also usable, that is, it can be used in the form of a sugar composition containing sophorose and / or laminaribiose. In this case, there are no particular restrictions on the content of sophorose and / or laminaribiose in the sugar composition used in the present invention, but for example, it is preferably 1% by mass or more per dry solids, more preferably 5% by mass or more, even more preferably 10% by mass or more, and most preferably 30% by mass or more. On the other hand, if the content of sophorose and / or laminaribiose per dry solids is less than 0.5% by mass, the functionality of sophorose and / or laminaribiose may not be fully exhibited. In this specification, "sophorose and / or laminaribiose content" means the total content of sophorose and laminaribiose, and if each is included individually, it means the content of sophorose or laminaribiose when included individually.

[0025] The sugar composition containing sophorose and / or laminaribiose described above may also contain other components such as sugars and oligosaccharides with different bonding modes and degrees of polymerization. Typically, for example, one or more oligosaccharides with a degree of polymerization of 2 to 10, such as sophorooligosaccharides (excluding sophorose), gentiooligosaccharides, cellooligosaccharides, and laminarioligosaccharides (excluding laminaribiose), can be exemplified by a composition containing each in the dry solid content equivalent shown below. Furthermore, as shown in the examples described later, the sugar composition may also contain β-glucooligosaccharides obtained by adding β-glucosidase to a glucose solution.

[0026] Sophorooligosaccharide: 5% by mass to 50% by mass Genthio-oligosaccharide: 5% to 80% by mass Cellooligosaccharides: 5% to 50% by mass Laminario oligosaccharide: 5% by mass to 50% by mass

[0027] As shown in the examples described later, sophorose and / or laminaribiose have the functionality to suppress the growth of bacteria in the vaginal flora that may have a negative impact on the vagina (also known as "bacteriostatic"), while promoting the growth of beneficial bacteria that have a positive impact on the vagina. Specifically, beneficial bacteria that have a positive impact on the vagina include, but are not limited to, Lactobacillus gasseri, Lactobacillus paragasseri, Lactobacillus crispatus, and Lactobacillus jensenii. On the other hand, bacteria that may have adverse effects on the vagina include Lactobacillus iners, Gardnerella vaginalis, Mobiluncus curtisii, and Atopobium vaginae.

[0028] Since the present invention uses sophorose and / or laminaribiose, which possess the above-described functionalities, as an active ingredient, it can be suitably used for the purpose of maintaining the balance of the vaginal flora and keeping the vagina in good condition.

[0029] Specifically, in one embodiment, the present invention can be suitably used for the purpose of improving the balance of the vaginal microbiota. When the balance of the vaginal microbiota is disrupted, excessive proliferation of commensal bacteria and invasion and proliferation of other harmful bacteria occur, inducing various symptoms. The present invention can be suitably used to prevent such conditions from occurring, or to alleviate or suppress the severity of such symptoms. In this case, improving the balance may involve, for example, increasing the proportion of beneficial bacteria such as Lactobacillus species or decreasing the proportion of harmful bacteria. Lactobacillus bacteria have an antagonistic effect against pathogenic bacteria, such as Gardnerella vaginalis, and can inhibit the proliferation of such harmful bacteria. Therefore, by promoting the proliferation of Lactobacillus bacteria, they can function as balance regulators for the vaginal microbiota. Furthermore, improving the balance of the vaginal flora may not affect the growth of beneficial bacteria such as Lactobacillus, but rather suppress the growth of harmful bacteria.

[0030] In one embodiment, the present invention can be suitably used for the purpose of maintaining a slightly acidic environment in the vagina and preventing infection by other bacteria. For example, by promoting the growth of beneficial bacteria such as Lactobacillus, these bacteria produce lactic acid or antimicrobial peptides that suppress the growth of pathogenic bacteria, and thus can be suitably used for the above-mentioned purpose.

[0031] In one embodiment, the present invention can be suitably used for the purpose of preventing bacterial vaginosis (BV), or reducing or suppressing the severity of its symptoms. For example, by inhibiting the growth of Gardnerella vaginalis, the formation of biofilms caused by this bacterium can be prevented, thereby suppressing the inhibition of bacterial vaginosis (BV) treatment, and thus it can be suitably used for the purposes described above.

[0032] In one embodiment, the present invention can be suitably used for the purpose of suppressing the recurrence of bacterial vaginosis (BV). For example, Lactobacillus iners usually proliferate after a patient with bacterial vaginosis (BV) receives antibiotic treatment, but by suppressing this, it is possible to prevent the vagina from becoming an environment that is prone to the recurrence of bacterial vaginosis (BV), and therefore the present invention can be suitably used for the above-mentioned purpose.

[0033] (Vaginal composition) The vaginal microbiome balance improving agent provided by the present invention may be used in the form of various vaginal compositions such as cosmetics, topical pharmaceuticals, and quasi-drugs. Examples of vaginal compositions include solid preparations such as suppositories and capsules; semi-solid preparations such as creams, gels, and ointments; and liquid preparations such as liquids and lotions. Preferably, the present invention is provided in the form of a cream, gel, ointment, or suppository.

[0034] When the vaginal microbiota balance improving agent provided by the present invention is used in the form of a vaginal composition, the sophorose and / or laminaribiose, or the sugar composition containing sophorose and / or laminaribiose, may be prepared in their respective forms using appropriate pharmaceutically acceptable formulation carriers and components according to conventional methods as appropriate. Examples of such formulation carriers and components include water, other aqueous solvents, petrolatum, glycerin, glyceryl monostearate, glyceryl stearate, self-emulsifying glyceryl monostearate, corn starch, sodium carboxymethyl starch, crystalline cellulose, cellulose, lactose, D-sorbitol, stearic acid, magnesium stearate, palmitic acid, cetyl palmitate, tartaric acid, hard fat, octyldodecanol, paraffin, light liquid paraffin, gelling hydrocarbons, stearyl alcohol, cetanol, cetostearyl alcohol, sucrose fatty acid esters, diethanolamine, dimethylpolysiloxane, squalane, sodium bicarbonate, medium-chain triglyceride, propylene glycol, propylene glycol fatty acid esters, 1,3-butylene glycol, polyethylene glycol, polyoxyethylene cetyl ether, isopropyl myristate, octyldodecyl myristate, cetyl myristate, etc.Other ingredients include stabilizers such as dibutylhydroxytoluene, sodium edetate, sodium sulfite, and anhydrous sodium sulfite; thickeners such as xanthan gum, hydroxypropyl methylcellulose, hydroxypropylcellulose, and carboxyvinyl polymer; surfactants such as sorbitan stearate, sorbitan monostearate, and polyoxyl stearate; preservatives such as butylparaben, methylparaben, propylparaben, ethylparaben, sodium benzoate, and benzyl alcohol; and pH adjusters such as hydrochloric acid, sulfuric acid, and phosphorus. Examples of buffering agents include inorganic acids such as acids, polyphosphates, and boric acid; organic acids such as lactic acid, acetic acid, citric acid, tartaric acid, malic acid, succinic acid, oxalic acid, gluconic acid, fumaric acid, propionic acid, acetic acid, aspartic acid, epsilon-aminocaproic acid, glutamic acid, and aminoethylsulfonic acid; and inorganic bases such as sodium bicarbonate, sodium carbonate, potassium hydroxide, sodium hydroxide, calcium hydroxide, and magnesium hydroxide. Examples of buffering agents include boric acid buffers, phosphate buffers, carbonate buffers, citrate buffers, acetate buffers, epsilon-aminocaproic acid, aspartic acid, and aspartates. These pharmaceutical carriers and components can be processed according to the type and form of product to be added (for example, by arbitrarily selecting and combining processes such as crushing, milling, washing, hydrolysis, fermentation, purification, pressing, extraction, fractionation, filtration, drying, powdering, granulation, dissolution, sterilization, pH adjustment, deodorization, decolorization, etc.), and supplied by arbitrarily selecting from various materials.

[0035] Furthermore, by combining the various pharmacological effects of each known raw material, it is possible to create a product that offers not only the functionality of improving the balance of the vaginal microbiota, which is the objective of this invention, but also multifunctional effects.

[0036] In the vaginal microbiota balance improving agent or vaginal composition provided by the present invention, the content of sophorose and / or laminaribiose may be in the range of 0.01% to 100% by mass per dry solids. If the content per dry solids is less than 0.01% by mass, the functionality of sophorose and / or laminaribiose may not be fully exhibited. The content of sophorose and / or laminaribiose is more preferably 0.1% by mass or more, and even more preferably 0.3% by mass or more.

[0037] When the vaginal microbiota balance improving agent or vaginal composition provided by the present invention is in a liquid, gel, cream, or other form containing water or other medium, the content of sophorose and / or laminaribiose in such form may be set as appropriate and is not particularly limited, but is typically in the range of 0.01 mg / g to 400 mg / g (the denominator is the weight of the preparation), 0.1 mg / g to 100 mg / g, or 1 mg / g to 50 mg / g.

[0038] The vaginal microbiota balance improving agent or vaginal composition provided by the present invention, when applied to the vagina, may be administered in an appropriate amount, and is not particularly limited. For example, the amount of sophorose and / or laminaribiose may be in the range of 0.001g to 1g, 0.005g to 0.5g, or 0.02g to 0.05g. [Examples]

[0039] The present invention will be described in more detail below with reference to examples, but these examples are not intended to limit the scope of the present invention.

[0040] [1. Preparation of α-gluco-oligosaccharides] A glucoamylase enzyme preparation was added at a rate of 50 mg per gram of solids to a 75 (w / w)% glucose solution. The pH was adjusted to 5.0, and the mixture was maintained at 66°C. After 63 hours of reaction, 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-oligosaccharides.

[0041] Preparation of 2. β-Glucooligosaccharide To a 60 (w / w)% glucose solution, 9.3 mg of β-glucosidase enzyme preparation per 1 g of solid content was added, and after adjusting the pH to 6.6, it was maintained at 60 °C. After 72 hours of reaction, 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-glucooligosaccharide.

[0042] Preparation of 3. High-purity sophorose 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 each of a protein derived from Bifidobacterium scardovii having 1,2-β-oligoglucan phosphorylase activity of 19 mg / mL (the protein disclosed as "BSGP" in Patent No. 7025941) and 17 mg / mL sucrose phosphorylase (Sigma-Aldrich) were added and reacted at 30 °C. After 7 days, the enzyme was inactivated by boiling for 10 minutes, the reaction solution was appropriately concentrated, and applied to an activated carbon column to remove monosaccharides and disaccharides. To this, hydrochloric acid was added so that the final concentration became 85 mM, and acid decomposition was carried out by holding at 80 °C for 17 hours. The decomposed sample was subjected to HPLC under the following conditions, and the disaccharide fraction was recovered to obtain high-purity sophorose.

[0043] <HPLC conditions> Column: ULTRON PS-80NL (Showa Chemical Industry Co., Ltd.) Column temperature: 50 °C Eluent: Ultrapure water Flow rate: 0.9 mL / min Detector: Differential refractive index detector Analysis time: 20 minutes

[0044] Preparation of 4. High-purity laminaribiose 8.0 g of Cardlan (Mitsubishi Corporation Life Sciences Ltd.) and 720 mL of ultrapure water were mixed, 8 mL of 1 M sodium hydroxide was added, and the mixture was stirred 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, 80 mg of yeast cell wall lytic enzyme (「Zymolyase (registered trademark)」, Nacalai Tesque, Inc.) was added. Cysteine was added to a final concentration of 10 mM, and the reaction was allowed to proceed overnight at 37 °C with vigorous stirring. The enzyme was inactivated by heating at 80 °C for 10 minutes, and after cooling to room temperature, the supernatant was recovered by centrifugation. This was subjected to HPLC under the same conditions as for the preparation of high-purity sophorose, and the disaccharide fraction was recovered to obtain high-purity laminaribiose.

[0045] 〔5. Sugar composition analysis〕 Sugar composition analysis was performed using HPLC. The content of each degree of polymerization component was determined from the peak area of the obtained chromatogram. Analysis of α-glucooligosaccharides was carried out according to HPLC condition A, and analysis of β-glucooligosaccharides and high-purity samples was carried out according to HPLC condition A and HPLC condition B.

[0046] <HPLC condition A> Column: ULTRON PS-80NL (Showa Chemical Industry Co., Ltd.) Column temperature: 50 °C Eluent: Ultrapure water Flow rate: 0.9 mL / min Detector: Differential refractive index detector [[ID=二十一]] [[ID=二十二]]Analysis time: 20 minutes

[0047] <HPLC condition B> Column: HILICpak VG-50 4E (Showa Denko K.K.) Column temperature: 40 °C Eluent: Acetonitrile: Ultrapure water = 8:2 Flow rate: 0.6 mL / min Detector: Charged particle detector Analysis time: 60 minutes

[0048] Tables 1 and 2 show the results of the sugar composition analysis.

[0049] [Table 1]

[0050] As shown in Table 1, all of the gluco-oligosaccharides prepared above had a sugar composition mainly consisting of disaccharides. The β-gluco-oligosaccharide contained 12.4% by mass of sophorose and 10.5% by mass of laminaribiose.

[0051] [Table 2]

[0052] As shown in Table 2, the high-purity preparations of sophorose and laminariviose prepared above all had a purity of at least 90% by mass.

[0053] [Test Example 1] For each type of bacteria shown in Table 3 as a normal vaginal flora, we investigated the change in bacterial count when culturing them under appropriate culture conditions and adding the α-gluco-oligosaccharide or β-gluco-oligosaccharide prepared above to the culture medium.

[0054] [Table 3]

[0055] Specifically, glucose was added to a suitable culture medium to a final concentration of 0.5 (w / v)%, and each bacterial species was cultured overnight to obtain a pre-culture solution. α-gluco-oligosaccharide or β-gluco-oligosaccharide prepared above was added to 3 mL of fresh medium to a final concentration of 0.5 (w / v), and then 0.03 mL of the pre-culture solution was mixed in and cultured for 24 hours. The bacterial turbidity (OD660) after culturing was measured. The same test was also performed with glucose added as a carbohydrate to the culture medium, and this was used as a comparative control. The results are shown as relative values ​​(percentage compared to the sugar-free group) with the bacterial turbidity when sterile ultrapure water was added instead of carbohydrates set to 100%.

[0056] As a result, as shown in Figure 1, significant growth was observed in all bacterial species in the glucose-added group, indicating poor selectivity for specific species. Growth of L. iners was also observed in the α-gluco-oligosaccharide-added group. On the other hand, in the β-gluco-oligosaccharide-added group, growth was observed in L. crispatus, as well as L. paragasseri, L. jensenii, and L. gasseri, but almost no growth was observed in the other four species.

[0057] From the above, it has become clear that β-gluco-oligosaccharides, which are mainly composed of disaccharides, have the effect of suppressing the growth of bacteria harmful to the skin while promoting the growth of beneficial bacteria in the vagina (L. crispatus, L. paragasseri, L. jensenii, and L. gasseri).

[0058] [Test Example 2] The effects of sophorose and laminalibiose, β-glucooligosaccharides composed of disaccharides, on promoting the growth of L. crispatus, L. paragasseri, L. jensenii, and L. gasseri were evaluated. Specifically, the effects on promoting the growth of L. crispatus, L. paragasseri, L. jensenii, and L. gasseri were evaluated in the same manner as in Test Example 1, except that sophorose or laminalibiose, which were prepared to a high purity as described above, were used as the carbohydrate added to the culture medium.

[0059] As a result, as shown in Figure 2, sophorose and laminaribiose had the effect of promoting the growth of L. crispatus, L. paragasseri, L. jensenii, and L. gasseri.

[0060] [Test Example 3] The effects of using sophorose and laminaribiose, which were prepared to high purity as disaccharide β-glucooligosaccharides as described above, on promoting the growth of L. crispatus, L. paragasseri, L. jensenii, and L. gasseri were evaluated in the same manner as in Test Example 2, except that the amount added to the culture medium was set to a final concentration of 0.1 (w / v)% or 1.0 (w / v)%.

[0061] As a result, as shown in Figure 3, when a preparation containing high purity sophorose or laminaribiose was added to the culture medium to a final concentration of 0.1 (w / v)% and cultured, an effect promoting the growth of L. crispatus, L. paragasseri, L. jensenii, and L. gasseri was observed. Furthermore, at a final concentration of 1.0 (w / v)%, a greater growth effect was observed than that shown in Test Example 2 at a final concentration of 0.5 (w / v)%.

[0062] [Test Example 4] The effects of using sophorose or laminaribiose, prepared to high purity as described above, as a β-gluco-oligosaccharide composed of disaccharides, on promoting the growth of Lactobacillus iners, Gardnerella vaginalis, Mobiluncus curtisii, and Atopobium vaginae were evaluated in the same manner as in Test Example 1, except that the amount added to the culture medium was set to a final concentration of 0.1(w / v)%, 0.5(w / v)%, or 1.0(w / v)%.

[0063] As a result, as shown in Figure 4, neither the amount of sophorose or laminaribiose added to the preparations containing high purity showed any effect in promoting the growth of Lactobacillus iners, Gardnerella vaginalis, Mobiluncus curtisii, and Atopobium vaginae compared to the control group without added sugar (percentage compared to the sugar-free group).

[0064] Based on the above, and considering the results of Test Examples 2-4 together, it has become clear that sophorose and laminaribiose, which are β-glucooligosaccharides composed of disaccharides, have the effect of suppressing the growth of harmful bacteria in the vagina while promoting the growth of beneficial bacteria in the vagina.

[0065] (Prescription example 1) This is an example of a formulation in which sophorose or laminaribiose is used as an ointment to improve the balance of the vaginal flora according to the present invention.

[0066] (Raw material name) Content (mass%) Sophorose or laminaribiose 0.5% White petrolatum 25.0% Stearyl alcohol 20.0% Propylene glycol 12.0% Polyoxyethylene hydrogenated castor oil 60 4.0% Glyceryl monostearate 1.0% Purified water remainder

[0067] (Prescription example 2) This is an example of a formulation in which β-gluco-oligosaccharide is used in an ointment as an agent for improving the balance of the vaginal flora according to the present invention.

[0068] (Raw material name) Content (mass%) β-Gluco-oligosaccharide 1.0% White petrolatum 25.0% Stearyl alcohol 20.0% Propylene glycol 12.0% Polyoxyethylene hydrogenated castor oil 60 4.0% Glyceryl monostearate 1.0% Purified water remainder

[0069] (Prescription example 3) This is a formulation example in which sophorose or laminaribiose is used as a vaginal suppository as an agent for improving the balance of the vaginal flora according to the present invention.

[0070] (Raw material name) Content (mass%) Sophorose or laminaribiose 11.5% Hard fat 88.5%

[0071] (Prescription example 4) This is an example of a formulation in which β-gluco-oligosaccharide is used in a vaginal suppository as an agent for improving the balance of the vaginal flora according to the present invention.

[0072] (Raw material name) Content (mass%) β-Gluco-oligosaccharide 11.5% Hard fat 88.5%

[0073] (Prescription example 5) This is an example of a formulation in which sophorose or laminaribiose is used as a cream to improve the balance of the vaginal flora according to the present invention.

[0074] (Raw material name) Content (mass%) Sophorose or laminaribiose 1.0% Stearyl alcohol 8.0% Behenyl alcohol 2.0% Isopropyl palmitate 6.0% Polysorbate 60 2.0% Sorbitan stearate 1.0% Self-emulsifying glyceryl monostearate 0.3% 1,3-Butylene glycol 20.0% Sodium dihydrogen phosphate 0.3% Hydrochloric acid (appropriate amount) Sodium hydroxide (appropriate amount) Purified water remainder

[0075] (Prescription example 6) This is an example of a formulation in which β-gluco-oligosaccharide is used in a cream as an agent for improving the balance of the vaginal flora according to the present invention.

[0076] (Raw material name) Content (mass%) β-Gluco-oligosaccharide 2.0% Stearyl alcohol 8.0% Behenyl alcohol 2.0% Isopropyl palmitate 6.0% Polysorbate 60 2.0% Sorbitan stearate 1.0% Self-emulsifying glyceryl monostearate 0.3% 1,3-Butylene glycol 20.0% Sodium dihydrogen phosphate 0.3% Hydrochloric acid (appropriate amount) Sodium hydroxide (appropriate amount) Purified water remainder

Claims

1. A vaginal microbiome balance-improving agent containing sophorose and / or laminaribiose as active ingredients.

2. While stimulating the action of one or more species selected from the group consisting of Lactobacillus iners, Gardnerella vaginalis, Mobiluncus curtisii, and Atopobium vaginae, Lactobacillus gasseri, Lactobacillus paragasseri, and Lactobacillus crispatus A vaginal microbiota balance improving agent according to claim 1, which is for promoting the growth of one or more species selected from the group consisting of *Crispatus* and *Lactobacillus jensenii*.

3. The vaginal microbiota balance improving agent according to claim 1, wherein the sophorose and / or laminaribiose used as the active ingredient is in the form of a sugar composition containing sophorose and / or laminaribiose produced using a glycosyltransferase / condensation enzyme.

4. A vaginal composition containing a vaginal microbiome balance improving agent according to any one of claims 1 to 3.

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