Agent for improving the balance of normal vaginal flora and vaginal composition

Abstract

To provide an agent for improving the balance of normal vaginal flora, which is useful for maintaining good vaginal condition. [Solution] The agent for improving the balance of normal vaginal flora uses sophorose and / or laminaribiose. Sophorose and / or laminaribiose have the functionality to suppress the growth of bacteria that may have a negative effect on the vagina among normal vaginal flora, while promoting the growth of beneficial bacteria that have a positive effect on the vagina. This agent for improving the balance of normal vaginal flora can be suitably used for the purpose of maintaining the balance of normal vaginal flora and keeping the vagina in good condition.

Description

[Technical Field]

[0001] The present invention relates to an agent for improving the balance of normal vaginal flora and a vaginal composition using sophorose and / or laminaribiose. [Background technology]

[0002] The vaginal flora of a healthy human is dominated by Lactobacillus bacteria, which maintain an acidic environment due to the lactic acid produced by these bacteria. An imbalance in the intrauterine bacterial flora can lead to bacterial vaginosis (BV), which causes itching and inflammation. BV can lead to miscarriage or premature birth, and can also spread infection to other organs, resulting in serious complications. BV is estimated to affect one in three women, and is a disease with a high recurrence rate, even when treated with antibiotics.

[0003] Lactobacillus bacteria that form the human vaginal microbiota include Lactobacillus gasseri, Lactobacillus paragasseri, Lactobacillus crispatus, and Lactobacillus jensenii. These Lactobacillus bacteria produce D-lactic acid and bacteriocins, which help maintain vaginal health.

[0004] However, some vaginal flora are known to be harmful when overgrowth occurs. For example, Lactobacillus iners produces L-lactic acid, but this is not sufficient to inhibit bacterial vaginosis (BV), making it more likely to progress to bacterial vaginosis (BV). Gardnerella vaginalis has been observed to grow in patients with bacterial vaginosis (BV), forming biofilms and inhibiting BV treatment. Mobiluncus curtisii has been observed to grow in patients with bacterial vaginosis (BV) and is known to be resistant to metronidazole, a treatment for bacterial vaginosis (BV). Atopobium vaginae has also been observed to grow in patients with bacterial vaginosis (BV).

[0005] For these reasons, the normal vaginal flora is considered to be extremely important in maintaining good vaginal condition.

[0006] Regarding normal vaginal flora, for example, Patent Document 1 discloses a method for maintaining the pH value of vaginal secretions within the range of 3.5 to 4.5 by using a combination of a sugar and at least one of benzoic acid and its sodium salt. Patent Document 2 discloses a preparation for preventing and / or treating vaginal infectious diseases, which contains a bacteriostatic agent containing phenethyl alcohol as an active ingredient. Patent Document 3 discloses a lactic acid bacteria strain for preventing and / or treating vaginal infectious diseases. [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 Summary of the Invention [Problem to be solved by the invention]

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

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

[0010] In order to achieve the above-mentioned object, the present inventors conducted various studies and discovered sophorose and / or laminaribiose as materials that selectively promote the growth of bacteria that have a positive effect on the vagina, leading to the completion of the present invention.

[0011] That is, in a first aspect, the present invention provides an agent for improving the balance of normal vaginal flora, which contains sophorose and / or laminaribiose as an active ingredient.

[0012] In the present invention, the agent for improving the balance of normal vaginal flora is preferably one that promotes the growth of one or more species selected from the group consisting of Lactobacillus gasseri, Lactobacillus paragasseri, Lactobacillus crispatus, and Lactobacillus jensenii, while bacteriostatically inhibiting one or more species selected from the group consisting of Lactobacillus iners, Gardnerella vaginalis, Mobiluncus curtisii, and Atopobium vaginae.

[0013] Furthermore, the sophorose and / or laminaribiose as the active ingredient is preferably in the form of a composition containing sophorose and / or laminaribiose produced using a glycosyltransferase-condensing enzyme.

[0014] On the other hand, in a second aspect, the present invention provides a vaginal composition containing the above-mentioned agent for improving the balance of normal vaginal flora. [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 are normally present in the vagina and that may have a negative effect on the vagina, while promoting the growth of beneficial bacteria that have a positive effect on the vagina. [Brief explanation of the drawings]

[0016] [Figure 1] This is a graph showing the results of an investigation into the change in bacterial count when various bacteria, which are normal vaginal flora, were cultured under culture conditions appropriate for each species, and α-glucooligosaccharide or β-glucooligosaccharide was added to the culture medium in Test Example 1. [Figure 2] This is a graph showing the results of an investigation into the change in bacterial count when various vaginal flora (L. crispatus, L. paragasseri, L. jensenii, L. gasseri) were cultured under culture conditions appropriate for each species in Test Example 2, and when highly purified sophorose or laminaribiose was added to the culture medium, the results were examined. [Figure 3] This is a graph showing the results of Test Example 3, in which various vaginal flora (L. crispatus, L. paragasseri, L. jensenii, L. gasseri) were cultured under culture conditions appropriate for each species, and highly purified sophorose or laminaribiose was added to the culture medium at various final concentrations to examine the change in bacterial count. [Figure 4]This is a graph showing the results of Test Example 4, in which various bacteria (Lactobacillus iners, Gardnerella vaginalis, Mobiluncus curtisii, Atopobium vaginae) that are normal vaginal flora were cultured under culture conditions appropriate for each bacteria, and the changes in the number of bacteria were examined when highly purified sophorose or laminaribiose was added to the culture medium at various final concentrations. DETAILED DESCRIPTION OF THE INVENTION

[0017] The present invention will be described in more detail below. In this specification, when a numerical range is indicated with "to", it means a range from above to below, and both ends of the range are included, unless otherwise specified. Furthermore, upper and lower limits can be appropriately combined, and the resulting numerical range is also included in the disclosure of this specification.

[0018] The agent for improving the balance of normal vaginal flora according to the present invention uses sophorose and / or laminaribiose as its active ingredients.

[0019] Sophorose (also known as 2-O-β-D-glucopyranosyl-D-glucose) is a disaccharide formed by the β-1,2 bond between two glucose molecules. It was first discovered as a sugar component of the glycoside pigment in the pods of the legume Sophora japonica, but it rarely exists in nature as a free disaccharide, and is a sugar that occurs in very small amounts in nature. It is only known to exist in small amounts in acid hydrolysates of starch and in royal jelly.

[0020] Sophorose can be obtained by any method, including, but not limited to, enzymatic preparation using a transglycosylation / condensation reaction. Specifically, as described in detail in Japanese Patent Nos. 2750374 and 3020583, sophorose can be obtained by allowing microbial β-glucosidase to act on glucose and / or β-glucooligosaccharides and utilizing the condensation / transglycosylation reaction inherent to the β-glucosidase. Furthermore, as described in detail in Japanese Patent No. 7025941, sophorose can be obtained in higher yields by allowing 1,2-β-oligoglucan phosphorylase to act on α-glucose-1-phosphate and glucose.

[0021] Laminaribiose (also known as 3-O-β-D-glucopyranosyl-D-glucose) is a disaccharide consisting of two glucose molecules linked together in a β-1,3 bond, and is known as the minimum building block of β-1,3 glucans such as laminaran, curdlan, and pachyman.

[0022] Laminaribiose can be obtained by, but is not limited to, enzymatic preparation using a transglycosylation / condensation reaction. Specifically, as described in detail in Japanese Patent Publication Nos. 2750374 and 3020583, laminaribiose can be obtained by allowing microbial β-glucosidase to act on glucose and / or β-glucooligosaccharides and utilizing the condensation / transglycosylation reaction inherent in β-glucosidase. Furthermore, as described in detail in Japanese Patent Publication No. 2955590, laminaribiose can be easily obtained in high yields using sucrose as a raw material in the presence of phosphoric acid in combination with three or four enzymes: sucrose phosphorylase, glucose isomerase, and laminaribiose phosphorylase and / or β-1,3-oligoglucan phosphorylase.

[0023] These methods allow for the production of compositions containing sophorose and / or laminaribiose. The resulting compositions containing sophorose and / or laminaribiose can be subjected to treatments such as filtration, decolorization, deodorization, and desalting, as necessary. Furthermore, the compositions may be subjected to treatments such as extraction, centrifugation, crystallization, microbial assimilation, and membrane fractionation such as chromatographic fractionation using activated carbon, porous carriers, hydrophobic resins, hydrophilic resins, ion exchange resins, and adsorption resins, dialysis, and ultrafiltration. 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, or any form containing sophorose and / or laminaribiose can be used, i.e., in the form of a sugar composition containing sophorose and / or laminaribiose. In this case, the content of sophorose and / or laminaribiose in the sugar composition used in the present invention is not particularly limited, but is preferably 1% by mass or more, more preferably 5% by mass or more, even more preferably 10% by mass or more, and most preferably 30% by mass or more, based on the dry solid content. On the other hand, if the content of sophorose and / or laminaribiose based on the dry solid content is less than 0.5% by mass, the functionality of sophorose and / or laminaribiose may not be fully exhibited. In this specification, the term "content of sophorose and / or laminaribiose" means the total content of sophorose and laminaribiose, and when each is contained alone, it means the content of sophorose or laminaribiose when contained alone.

[0025] The sugar composition containing sophorose and / or laminaribiose may also contain other components, such as sugars or oligosaccharides with different linkages or degrees of polymerization. Typical examples include those containing one or more oligosaccharides with degrees of polymerization of 2 to 10, such as sophorooligosaccharides (excluding sophorose), gentiooligosaccharides, cellooligosaccharides, and laminarioligosaccharides (excluding laminaribiose), each in the amount calculated on a dry solid content basis as shown below. Furthermore, as shown in the Examples below, the sugar composition may also contain β-glucooligosaccharides obtained by adding β-glucosidase to a glucose solution.

[0026] Sophorooligosaccharides: 5% 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 below, sophorose and / or laminaribiose have the functionality to inhibit the growth of bacteria that may have a negative effect on the vagina among the normal vaginal flora (also known as "bacteriostatic"), while promoting the growth of beneficial bacteria that have a positive effect on the vagina. Specific examples of beneficial bacteria that have a positive effect 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] The present invention uses sophorose and / or laminaribiose, which have the above-mentioned functionality, as active ingredients, and can therefore be suitably used for the purpose of maintaining the balance of normal vaginal flora and keeping the vagina in good condition.

[0029] Specifically, in one embodiment, the present invention can be suitably used to improve the balance of normal vaginal flora. Disruption of the balance of normal vaginal flora can lead to excessive proliferation of normal bacteria and the invasion and proliferation of other harmful bacteria, resulting in various symptoms. The present invention can be suitably used to prevent such conditions or to alleviate or suppress the severity of these symptoms. In this case, improving the balance can be achieved, for example, by increasing the proportion of beneficial bacteria, such as Lactobacillus, or by decreasing the proportion of harmful bacteria. Lactobacillus bacteria have an antagonistic effect against pathogenic bacteria, such as Gardnerella vaginalis, and can inhibit the growth of such harmful bacteria. Therefore, by promoting the growth of Lactobacillus bacteria, they can function as a balance adjuster for normal vaginal flora. Furthermore, improving the balance of normal vaginal flora may involve, for example, suppressing the growth of harmful bacteria without affecting the growth of beneficial bacteria such as Lactobacillus.

[0030] In one embodiment, the present invention can be suitably used to maintain a slightly acidic vagina and prevent infection by other bacteria. For example, by promoting the growth of beneficial bacteria such as Lactobacillus, these bacteria produce lactic acid and antimicrobial peptides that suppress the growth of pathogenic bacteria, and therefore can be suitably used for the above-mentioned purposes.

[0031] In some embodiments, 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 a biofilm caused by this bacterium can be prevented, thereby suppressing the inhibition of bacterial vaginosis (BV) treatment, and therefore the present invention can be suitably used for the purpose described above.

[0032] In one aspect, the present invention can be suitably used for the purpose of suppressing the recurrence of bacterial vaginosis (BV). For example, Lactobacillus iners usually proliferates after a patient with bacterial vaginosis (BV) is treated with antibiotics. By inhibiting this proliferation, the vaginal environment can be prevented from becoming prone to the recurrence of bacterial vaginosis (BV), and therefore the present invention can be suitably used for the purpose described above.

[0033] (Vaginal composition) The vaginal flora balance improving agent provided by the present invention may be used in the form of various vaginal compositions, such as cosmetics, external medicines, and quasi-drugs. Examples of vaginal compositions include solid formulations such as suppositories and capsules; semi-solid formulations such as creams, gels, and ointments; and liquid formulations such as liquids and lotions. The preferred forms of the present invention are creams, gels, ointments, and suppositories.

[0034] When the agent for improving the balance of vaginal resident flora provided by the present invention is used in the form of a vaginal composition, the above-mentioned sophorose and / or laminaribiose, or a sugar composition containing sophorose and / or laminaribiose, can be prepared into the respective forms using appropriate pharmaceutically acceptable carriers and ingredients according to conventional methods. Examples of such pharmaceutical 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, cetyl alcohol, cetostearyl alcohol, sucrose fatty acid esters, diethanolamine, dimethylpolysiloxane, squalane, sodium bicarbonate, medium-chain fatty acid triglycerides, propylene glycol, propylene glycol fatty acid esters, 1,3-butylene glycol, polyethylene glycol, polyoxyethylene cetyl ether, isopropyl myristate, octyldodecyl myristate, cetyl myristate, and the like.Other ingredients include stabilizers such as dibutylhydroxytoluene, sodium edetate, sodium sulfite, and dried sodium sulfite; thickeners such as xanthan gum, hydroxypropylmethylcellulose, 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. inorganic 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, aminoethylsulfonic acid, and other organic acids; inorganic bases such as sodium bicarbonate, sodium carbonate, potassium hydroxide, sodium hydroxide, calcium hydroxide, magnesium hydroxide, and other buffers such as borate buffer, phosphate buffer, carbonate buffer, citrate buffer, acetate buffer, epsilon-aminocaproic acid, aspartic acid, and aspartate salts. These pharmaceutical carriers and ingredients may be selected from a variety of materials and provided after undergoing conventional processing (e.g., grinding, milling, washing, hydrolysis, fermentation, refining, pressing, extraction, fractionation, filtration, drying, powdering, granulation, dissolution, sterilization, pH adjustment, deodorization, decolorization, etc., optionally selected and combined) according to the type and form of the product to which they are to be added.

[0035] Furthermore, by combining the various medicinal effects of each raw material known to date, it is possible to create a product that is expected to have multifunctional effects in addition to the functionality of improving the balance of the normal vaginal flora, which is the objective of this invention.

[0036] In the vaginal flora balance improving agent or vaginal composition provided by the present invention, the content of sophorose and / or laminaribiose can be in the range of 0.01% to 100% by mass based on the dry solid content. If the content based on the dry solid content is less than 0.01% by mass, the functionality of sophorose and / or laminaribiose may not be fully exerted. 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 flora balance improving agent or vaginal composition provided by the present invention is in a form containing water or another medium, such as a liquid, gel, or cream, the content of sophorose and / or laminaribiose in such a form may be set appropriately and is not particularly limited, but may typically be in the range of 0.01 mg / g to 400 mg / g (the denominator indicates the weight of the preparation), 0.1 mg / g to 100 mg / g, or 1 mg / g to 50 mg / g.

[0038] When the vaginal flora balance improving agent or vaginal composition provided by the present invention is applied to the vagina, the amount to be administered may be appropriately determined and is not particularly limited, and may be, for example, in the range of 0.001 g to 1 g, 0.005 g to 0.5 g, or 0.02 g to 0.05 g of sophorose and / or laminaribiose. [Example]

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

[0040] 1. Preparation of α-Glucooligosaccharides A glucoamylase enzyme preparation was added to a 75 (w / w)% glucose solution at a rate of 50 mg per 1 g of solids, and the solution was adjusted to pH 5.0 and maintained at 66°C. After 63 hours of reaction, the enzyme was inactivated by adjusting the pH to 4.0 and raising the temperature to 80°C. Monosaccharides were removed from the product by membrane fractionation to obtain α-glucooligosaccharides.

[0041] Preparation of 2. β - Glucooligosaccharide A β - glucosidase enzyme preparation was added to a 60 (w / w)% glucose solution at 9.3 mg per 1 g of solid content, 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, and 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 (protein disclosed as "BSGP" in Patent No. 7025941) at 19 mg / mL and 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 was subjected to an activated carbon column to remove monosaccharides and disaccharides. To this, hydrochloric acid was added to a final concentration of 85 mM and maintained at 80°C for 17 hours for acid hydrolysis. The decomposed sample was subjected to HPLC under the following conditions, and the disaccharide fraction was collected 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 mixture was centrifuged and the supernatant was recovered. 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. The analysis of α-glucooligosaccharides was carried out according to HPLC condition A, and the 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 Denko K.K.) Column temperature: 50 °C Eluent: Ultrapure water Flow rate: 0.9 mL / min Detector: Differential refractive index detector 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 glucooligosaccharides prepared above had a sugar composition mainly composed of disaccharides. The β-glucooligosaccharides 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 laminaribiose prepared above all had a purity of at least 90% by mass.

[0053] [Test Example 1] The various bacteria shown in Table 3, which are normal vaginal flora, were cultured under appropriate culture conditions, and the changes in bacterial counts were examined when the α-glucooligosaccharides or β-glucooligosaccharides prepared above were added to the culture medium.

[0054] [Table 3]

[0055] Specifically, glucose was added to the appropriate medium to a final concentration of 0.5 (w / v)%. Each bacterial species was cultured overnight to prepare a preculture solution. The α-glucooligosaccharide or β-glucooligosaccharide prepared above was added to 3 mL of fresh medium to a final concentration of 0.5 (w / v)%. 0.03 mL of the preculture solution was then added and cultured for 24 hours. The bacterial turbidity (OD660) after culture was measured. A similar test was also performed using the medium supplemented with glucose as a carbohydrate, as a control. The results are shown as a relative value (% of the control without added sugar), with the bacterial turbidity when sterilized ultrapure water was added instead of carbohydrate being taken as 100%.

[0056] As shown in Figure 1, the glucose-supplemented medium showed significant growth of all bacterial species, with little selectivity for specific bacterial species. Growth of L. iners was also observed in the α-glucooligosaccharide-supplemented medium. On the other hand, growth of L. crispatus, L. paragasseri, L. jensenii, and L. gasseri was observed in the β-glucooligosaccharide-supplemented medium, but the other four bacterial species showed little growth.

[0057] From the above, it has become clear that β-glucooligosaccharides, 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] Sophorose and laminaribiose, which are disaccharide β-glucooligosaccharides, were evaluated for their growth-promoting effects on L. crispatus, L. paragasseri, L. jensenii, and L. gasseri. Specifically, the growth-promoting effects on L. crispatus, L. paragasseri, L. jensenii, and L. gasseri were evaluated in the same manner as in Test Example 1, except that the highly purified sophorose or laminaribiose prepared above was used as the carbohydrate added to the 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 sophorose and laminaribiose prepared to high purity as described above were used as disaccharide β-glucooligosaccharides, and the effects of 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 they were added to the medium at 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 highly purified sophorose or laminaribiose was added to a medium at a final concentration of 0.1 (w / v)%, the growth of L. crispatus, L. paragasseri, L. jensenii, and L. gasseri was promoted. Furthermore, at a final concentration of 1.0 (w / v), a greater growth promotion effect was observed than the growth promotion effect of L. crispatus, L. paragasseri, L. jensenii, and L. gasseri shown at a final concentration of 0.5 (w / v)% in Test Example 2.

[0062] [Test Example 4] The sophorose or laminaribiose prepared to high purity as described above was used as the disaccharide β-glucooligosaccharide, and the effect of promoting the growth of Lactobacillus iners, Gardnerella vaginalis, Mobiluncus curtisii, and Atopobium vaginae was evaluated in the same manner as in Test Example 1, except that the amount added to the 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, no effect of promoting the growth of Lactobacillus iners, Gardnerella vaginalis, Mobiluncus curtisii, or Atopobium vaginae was observed, regardless of the amount of preparation containing highly purified sophorose or laminaribiose added, compared to the relative value to the control with no added sugar (% of the no sugar added group).

[0064] Considering the results of Test Examples 2 to 4, it has become clear that sophorose and laminaribiose, which are disaccharide β-glucooligosaccharides, have the effect of suppressing the growth of bacteria harmful to the vagina while promoting the growth of bacteria beneficial to the vagina.

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

[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 a formulation example in which β-glucooligosaccharide is used as an agent for improving the balance of normal vaginal flora according to the present invention in the form of an ointment.

[0068] (Raw material name) Content (mass%) β-glucooligosaccharides 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 normal 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 a formulation example in which β-glucooligosaccharide is used as a vaginal suppository as an agent for improving the balance of normal vaginal flora according to the present invention.

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

[0073] (Prescription Example 5) This is a formulation example in which sophorose or laminaribiose is used as a cream as an agent for improving the balance of normal 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 a formulation example in which β-glucooligosaccharide is used as a cream formulation as an agent for improving the balance of normal vaginal flora according to the present invention.

[0076] (Raw material name) Content (mass%) β-glucooligosaccharides 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. 1. An agent for improving the balance of normal vaginal flora, which contains sophorose and / or laminaribiose as active ingredients, and which is to be applied to the vagina.

2. The present invention relates to a method for producing a microbial colony containing Lactobacillus iners, Gardnerella vaginalis, Mobiluncus curtisii, and Atopobium vaginae, and a method for producing the same, wherein the microbial colony containing Lactobacillus gasseri, Lactobacillus paragasseri, Lactobacillus crispatus, and the like are subjected to bacteriostatic action against one or more bacteria selected from the group consisting of Lactobacillus iners, Gardnerella vaginalis, Mobiluncus curtisii, and Atopobium vaginae.

2. The vaginal flora balance improving agent according to claim 1, which is for promoting the growth of one or more species selected from the group consisting of Lactobacillus crispatus and Lactobacillus jensenii.

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

4. A vaginal composition for improving the balance of normal vaginal flora to be applied to the vagina, comprising the agent for improving the balance of normal vaginal flora according to any one of claims 1 to 3.

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