Agent for inhibiting bacterial proliferation, and cosmetic, quasi-medicinal and pharmaceutical composition containing said agent, and method for inhibiting bacterial proliferation

Abstract

The present invention relates to an agent for suppressing the proliferation of at least one selected bacterium from the group comprising the genera Gardnerella and Atopobium. The agent contains glyceryl glucoside.

Description

Bacterial growth inhibitor, cosmetics, quasi-drugs, and pharmaceutical compositions containing the same, and method for inhibiting bacterial growth

[0001] The present disclosure relates to an inhibitor of the growth of at least one bacterium selected from the group consisting of the genus Gardnerella and the genus Atopobium, a cosmetic, a quasi-drug, and a pharmaceutical composition containing the same, and a method for inhibiting bacterial growth.

[0002] Various bacteria are present in the vagina of healthy adult women, and Lactobacillus species account for 75-95% of these. Lactobacillus species (Döderlein's bacillus) that are normally present in the vagina produce lactic acid and maintain an acidic pH of 4.5 or less, thereby preventing the invasion of other bacteria (Non-Patent Document 1).

[0003] Known Döderlein's bacilli include Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus jensenii, Lactobacillus casei, Lactobacillus vaginalis, Lactobacillus crispatus, and Lactobacillus iners. Among these, Lactobacillus crispatus and Lactobacillus iners have been reported to be predominant bacteria in all four ethnic groups (Caucasians, Blacks, Hispanics, and Asians) (Non-Patent Document 2).

[0004] Vaginitis (vaginosis) is a disease concept in which abnormal vaginal discharge is the main complaint, and representative examples include bacterial vaginosis (BV), genital candidiasis, and vaginal trichomoniasis. Genital candidiasis is caused by fungi of the genus Candida, and vaginal trichomoniasis is caused by the protozoan Trichomonas vaginalis. However, bacterial vaginosis occurs due to the disruption of normal flora and is not caused by a specific causative microorganism.

[0005] In bacterial vaginosis, the number of Doderlein's bacilli is reduced, and various aerobic and / or anaerobic bacteria are abnormally proliferating. Although more than half of patients with bacterial vaginosis are asymptomatic, the main symptoms of bacterial vaginosis are foul-smelling vaginal discharge, gray to white vaginal discharge, burning sensation during urination, itching around the vagina, etc., and bacterial vaginosis is thought to increase the risk of pelvic inflammatory disease, miscarriage, premature birth, postpartum endometritis, etc.

[0006] In bacterial vaginosis, known aerobic bacteria that overgrow include Streptococcus agalactiae, Escherichia coli, Gardnella vaginalis, etc., and known anaerobic bacteria that overgrow include Atopobium spp. such as Anaerocossus spp. and Atopobium vaginae, Peptostreptococcus spp., Finegoldia spp., Micromonas spp., Peptoniphilus spp., etc.

[0007] Metronidazole is known as a drug that has a bacterial growth inhibitory effect, mainly targeting Gardnella vaginalis, and is widely used to treat bacterial vaginosis (Non-Patent Document 1). However, bacteria of the genus Atopobium are known to be resistant to metronidazole (Patent Document 1), and the effect of metronidazole is insufficient.

[0008] On the other hand, glyceryl glycoside is a sugar derivative with a structure in which glucose is bound to glycerin, and has been used as a cosmetic raw material. α-D-Glucopyranosylglycerol, a type of glyceryl glucoside, is non-irritating to the skin and is extremely safe. It has also been reported to have antibacterial properties against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Pseudomonas cepacia, Staphylococcus epidermidis, and Corynebacterium minutissimum (Patent Document 2).

[0009] However, in Patent Document 2, the antibacterial activity is evaluated by inoculating bacteria into α-D-glucopyranosylglycerol, exposing them for 24 or 48 hours, collecting samples, culturing them in a medium, and counting the number of viable bacteria, and Patent Document 2 does not disclose how α-D-glucopyranosylglycerol acts on growing bacteria. Furthermore, it is not known how α-D-glucopyranosylglycerol acts on other bacteria, particularly bacteria of the genera Lactobacillus, Gardnerella, and Atopobium.

[0010] JP-T-2012-524117 A JP-A-2004-331577 A

[0011] Bacterial vaginosis, Sexually Transmitted Diseases Diagnosis and Treatment Guidelines 2008, Japanese Society for Sexually Transmitted Diseases, pp. 77-80, November 30, 2008. "Vaginal microbiome of reproductive-age women", Jacques Ravel et al., Proc Natl Acad Sci USA (2011), 108(Suppl 1); 4680-4687

[0012] In light of the above background, the present inventors have considered that if it were possible to reduce the number of bacteria belonging to the genus Gardnerella and Atopobium without reducing the number of Döderlein's bacillus, this would be preferable in terms of maintaining the health of areas where at least one bacterium selected from the group consisting of the genus Gardnerella and Atopobium is present or suspected to be present, such as delicate areas, and preventing or treating infections.

[0013] Therefore, an object of the present disclosure is to provide a bacterial growth inhibitor that does not inhibit the growth of Döderlein's bacillus but inhibits the growth of at least one bacterium selected from the group consisting of the genus Gardnerella and the genus Atopobium, a formulation containing the same, and a method for inhibiting bacterial growth.

[0014] The present inventors have conducted extensive research to solve the above-mentioned problems. As a result, they have found that the above-mentioned problems can be solved by having the following configuration, and have completed the present invention. The present invention relates to, for example, the following [1] to [7]. [1] A growth inhibitor for at least one bacterium selected from the group consisting of the genus Gardnerella and the genus Atopobium, comprising glyceryl glucoside. [2] The growth inhibitor according to [1], wherein the bacterium is at least one selected from the group consisting of Gardnerella vaginalis and Atopobium vaginae. [3] The growth inhibitor according to [1] or [2], which is used as a cleanser or external preparation. [4] A cosmetic or quasi-drug, comprising the growth inhibitor according to [1] or [2]. [5] A pharmaceutical composition for treating or preventing a disease associated with Gardnerella vaginalis or Atopobium vaginae, comprising the growth inhibitor according to [2]. [6] The pharmaceutical composition according to [5], wherein the disease is bacterial vaginosis. [7] A method for inhibiting the growth of at least one bacterium selected from the group consisting of the genus Gardnerella and the genus Atopobium, comprising the step of administering glyceryl glucoside to a subject.

[0015] According to the present disclosure, it is possible to provide a bacterial growth inhibitor that does not inhibit the growth of Döderlein's bacillus but inhibits the growth of at least one bacterium selected from the group consisting of the genus Gardnerella and the genus Atopobium, a formulation containing the same, and a method for inhibiting bacterial growth.

[0016] Glyceryl glucoside is highly safe, as no toxicity has been observed in any of the following tests: primary skin irritation test (rabbit), continuous skin irritation test, skin sensitization test (guinea pig intradermal maximization method), phototoxicity test (guinea pig), photosensitization test, eye irritation test (rabbit), mutagenicity test (in vitro), and human patch test. Therefore, according to the present disclosure, the growth of at least one bacterium selected from the group consisting of the genus Gardnerella and the genus Atopobium can be inhibited without concerns about safety or side effects.

[0017] Figure 1 is a graph showing the actual measured values ​​of turbidity in a bacterial growth test (mean values ​​of n=3). Figure 2 is a graph showing the relative values ​​of turbidity in a bacterial growth test to the control (mean values ​​of n=3).

[0018] The present invention will now be described in detail. The expression "A to B" regarding a numerical range means A or more and B or less unless otherwise specified. Furthermore, % means mass %.

[0019] <Glyceryl glucoside> In this specification, glyceryl glucoside, also referred to as GG, refers to a compound formed by α- or β-glucosidic bonding of glycerin with one or more glucose molecules. Examples of glyceryl glucoside include compounds represented by formulas (1) to (5). These compounds may be used singly or in combination of two or more.

[0020]

[0021]

[0022] The compound represented by formula (1) is 1-O-(α- or β-)D-(mono- or poly-)glucopyranosylglycerol, which is also called 1-O-(α- or β-)D-dihydroxypropyl (mono- or poly-)glucopyranoside.

[0023] The carbon atom at the 2-position marked with an asterisk in formula (1) is an asymmetric carbon atom, and optical isomers, i.e., a (2R) isomer and a (2S) isomer, exist. n in formula (1) represents the degree of sugar condensation and represents an integer of 1 or more. In commercially available glyceryl glucosides produced by known methods such as enzymatic methods and organic synthesis methods, n is usually an integer of 1 to 6, with most being an integer of 1 to 3, and compounds in which n is 1 are particularly abundant in GG compositions.

[0024] The compound represented by formula (2) is 2-O-(α- or β-)D-(mono- or poly-)glucopyranosylglycerol, which is also called 2-O-(α- or β-)D-dihydroxypropyl (mono- or poly-)glucopyranoside.

[0025] In formula (2), n represents the degree of sugar condensation and is an integer of 1 or greater. In commercially available glyceryl glucosides produced by known methods such as enzymatic methods and organic synthesis, n is usually an integer of 1 to 6, with most glyceryl glucosides being an integer of 1 to 3. In particular, compounds with n=1 are abundant in GG compositions.

[0026] m and n in formula (3), n and l in formula (4), and l, m, and n in formula (5) are sugar condensation degrees, each independently representing an integer of 1 or more, usually an integer of 1 to 6, mostly an integer of 1 to 3, and particularly compounds in which n, m, and l are 1 are abundant in GG compositions. In formulas (3), (4), and (5), when n, m, and l are 2 or more, the glucopyranosyl groups may be linked to each other via an α1-4 bond, a β1-4 bond, an α1-6 bond, a β1-6 bond, or the like.

[0027] The glyceryl glucoside in the present invention is represented by formula (1) or formula (2), and preferably contains at least one of 1-O-D-glucopyranosylglycerol and 2-O-D-glucopyranosylglycerol, where n is 1. More specifically, it preferably contains at least one selected from 1-O-α-D-glucopyranosylglycerol, 1-O-β-D-glucopyranosylglycerol, 2-O-α-D-glucopyranosylglycerol, and 2-O-β-D-glucopyranosylglycerol.

[0028] The glyceryl glucoside in the present invention may be one compound selected from 1-O-α-D-glucopyranosylglycerol, 1-O-β-D-glucopyranosylglycerol, 2-O-α-D-glucopyranosylglycerol, and 2-O-β-D-glucopyranosylglycerol, or may contain two or more compounds.

[0029] Furthermore, the glyceryl glucosides of 1-O-α-D-glucopyranosylglycerol, 1-O-β-D-glucopyranosylglycerol, 2-O-α-D-glucopyranosylglycerol, and 2-O-β-D-glucopyranosylglycerol are formed by binding glycerin to one glucose molecule. However, in the present invention, the glyceryl glucoside may be one to which an additional glucose molecule is bound, i.e., one in which n is 2 or greater, or may be a mixture of a compound in which n is 1 and a compound in which n is 2 or greater.

[0030] For example, in formulas (1) to (5), when n, m, and 1 are each independently 2 or greater, the bonding mode between glucopyranosyl groups is not particularly limited. Note that the wavy lines in formulas (1) to (5) each independently indicate that the hydroxyl group of glycerin may bond to the hydroxyl group at position 1 of the α-D-glucopyranosyl group by reacting with the hydroxyl group at position 1 of the α-D-glucopyranosyl group (α-bond) or may bond to the hydroxyl group at position 1 of the β-D-glucopyranosyl group by reacting with the hydroxyl group at position 1 of the β-D-glucopyranosyl group (β-bond).

[0031] The glyceryl glucoside of the present invention preferably has a purity (the mass proportion of components in glyceryl glucoside where n = 1 to 3 in formula (1) and formula (2), n + m = 2 or 3 in formula (3), n + l = 2 or 3 in formula (4), and n = m = l = 1 in formula (5)) of 70 to 100% by mass relative to 100% by mass of glyceryl glucoside, more preferably 60 to 95% by mass, and even more preferably 75 to 90% by mass. Furthermore, the glyceryl glucoside preferably contains 1-O-α-D-glucopyranosylglycerol as the main component, more preferably 1-O-α-D-glucopyranosylglycerol in an amount of 10 to 50% by mass relative to 100% by mass of glyceryl glucoside.

[0032] Glyceryl glucoside may be used as a composition (also referred to as a GG composition) that further contains, in addition to the compounds represented by formulas (1) to (5), compounds other than the compounds represented by formulas (1) to (5), such as water, unreacted substances or reaction by-products (impurities) that may be mixed in when preparing the compounds represented by formulas (1) to (5) by organic synthesis.

[0033] The impurities include glycerin and reaction by-products derived therefrom, which are used as raw materials for producing glyceryl glucoside, as well as glucose sources and reaction by-products derived therefrom, such as alcohols such as ethanol, isopropyl alcohol, ethylene glycol, diethylene glycol, triethylene glycol, propylene glycol, dipropylene glycol, 1,3-butylene glycol, 1,2-pentanediol, glycerin, diglycerin, and polyglycerin, and sugars such as glucose.

[0034] Incidentally, the amount of glycerin that can be incorporated into foods and beverages, seasonings, cosmetics, etc. may be limited. Therefore, when the GG composition is used to manufacture such products, the proportion of glycerin remaining in the GG composition is preferably 70% by mass or less, more preferably 50% by mass or less, even more preferably 45% by mass or less, particularly preferably 10% by mass or less, and most preferably 5% by mass or less, relative to the total solid content (components excluding water) in the GG composition. It is preferable that no glycerin is contained, and the proportion may be 0% by mass or less, and is usually 10% by mass or less, relative to the total solid content in the GG composition.

[0035] The method for producing glyceryl glucoside in the present invention is not particularly limited, and it can be produced by known methods such as an enzymatic method, an organic synthesis method, etc. Alternatively, commercially available products containing glyceryl glucoside can be purchased and used, for example, "COSARTE-2G (registered trademark) (manufactured by Toyo Sugar Refining Co., Ltd.)" and "C-mo 2G (manufactured by Toyo Sugar Refining Co., Ltd.)" can be used.

[0036] "COSARTE-2G" has a glyceryl glucoside content of approximately 66 to 78% by mass and a glycerin content of approximately 10% by mass or less, based on 100% by mass of "COSARTE-2G." "C-mo 2G" has a glyceryl glucoside content of approximately 30 to 38% by mass and a glycerin content of approximately 36 to 44% by mass, based on 100% by mass of "C-mo 2G."

[0037] Glyceryl glucoside has good water solubility, so it can be uniformly dissolved or dispersed even when added to water or preparations with a high moisture content. Furthermore, glyceryl glucoside has a high moisturizing effect and is less irritating to the skin, so it can be suitably used in sensitive areas such as delicate zones.

[0038] <Growth Inhibitor> A growth inhibitor according to one embodiment of the present invention contains glyceryl glucoside and inhibits the growth of at least one bacterium selected from the group consisting of the genus Gardnerella and the genus Atopobium.

[0039] The amount of glyceryl glucoside contained in the growth inhibitor is not particularly limited. For example, the lower limit of the amount of glyceryl glucoside contained in the growth inhibitor can be, for example, 20% by mass, 25% by mass, 30% by mass, 40% by mass, 45% by mass, 50% by mass, 60% by mass, 70% by mass, or 80% by mass. Furthermore, the upper limit of the amount of glyceryl glucoside contained in the growth inhibitor can be, for example, 100% by mass, 99% by mass, 98% by mass, 95% by mass, 90% by mass, or 85% by mass. The range of the amount of glyceryl glucoside contained in the growth inhibitor can be arbitrarily set by combining the lower and upper limits, for example, 25 to 100% by mass, 30 to 90% by mass, 60 to 80% by mass, or the like. From the viewpoint of the growth inhibitory effect, the amount of glyceryl glucoside contained in the growth inhibitor is preferably 20% by mass or more, more preferably 30% by mass or more.

[0040] The growth inhibitor may contain glycerin, diglycerin, glucose, water, and other components as needed.

[0041] Examples of bacteria of the genus Gardnerella include Gardnerella vaginalis, and examples of bacteria of the genus Atopobium include Atopobium vaginae (Fannyhessea vaginae), Atopobium rimae, Atopobium parvulum, Atopobium fossor, Atopobium minutum, and Atopobium deltae. Since these bacteria are the main bacteria that overgrow in bacterial vaginosis, it is preferable that the bacterium be at least one selected from the group consisting of Gardnerella vaginalis and Atopobium vaginae.

[0042] The method for evaluating the effect of inhibiting bacterial growth is not particularly limited, and any known method can be used. For example, if the degree of growth of bacteria cultured with the addition of a test substance is lower than the degree of growth of bacteria cultured without the addition of the test substance under the same culture conditions, the test substance can be evaluated as having an effect of inhibiting bacterial growth.

[0043] More specifically, as will be described later in the Examples, when the number of bacteria cultured without the addition of a test substance is taken as 100%, if the number of bacteria cultured with the addition of a test substance is less than 100%, the test substance can be evaluated as having an inhibitory effect on bacterial growth. In this case, the number of bacteria cultured with the addition of a test substance is preferably 90% or less, more preferably 80% or less, even more preferably 70% or less, and particularly preferably 60% or less of the number of bacteria cultured without the addition of the test substance.

[0044] The bacteria used are preferably growing bacteria, more preferably bacteria in the logarithmic growth phase.

[0045] The method for measuring the bacterial count is not particularly limited, and known methods can be used. The bacterial count may be measured by direct detection methods that count the number of bacteria, such as counting with a bacterial counting chamber, culturing on an agar plate and counting colonies, or culturing on an agar plate and observing the presence and size of growth inhibition zones formed, or by indirect methods that quantitatively detect various indicators attributable to the presence of bacteria, such as measuring turbidity (absorbance) using a spectrophotometer, measuring the weight of bacterial bodies, or measuring bacterial components or biochemical activity. Because it is simple and easy, measuring the bacterial count by an indirect method is preferred, and measuring turbidity using a spectrophotometer is more preferred.

[0046] <Uses of the growth inhibitor> The uses of the growth inhibitor of the present invention are not particularly limited, but since the growth inhibitor has a high effect of inhibiting the growth of at least one bacterium selected from the group consisting of the Gardnerella and Atopobium genera, it can be administered to a living body once or multiple times, for example, for the purpose of maintaining the health of delicate areas, etc. and / or for the purpose of preventing or treating infectious diseases.

[0047] The growth inhibitor is useful for preventing or ameliorating a disease or symptom associated with at least one bacterium selected from the group consisting of the genus Gardnerella and the genus Atopobium, and contributes to preventing or ameliorating the onset and progression of, for example, bacterial vaginosis, urinary tract infection, endometritis, acute salpingitis, postoperative infection, amniotic fluid infection, infertility, premature birth, neonatal sepsis, etc. The disease or symptom is preferably bacterial vaginosis.

[0048] The dosage of the growth inhibitor may be appropriately selected depending on the type or severity of the disease or symptom associated with at least one bacterium selected from the group consisting of the genus Gardnerella and the genus Atopobium, the route of administration, and the like.

[0049] The administration period of the growth inhibitor may be appropriately selected depending on the type or severity of the disease or symptom associated with at least one bacterium selected from the group consisting of the genus Gardnerella and the genus Atopobium, the administration route, etc. From the viewpoint of the growth inhibitory effect, the administration period is preferably 3 to 80 days, more preferably 7 to 60 days.

[0050] The route of administration of the growth inhibitor is not particularly limited, and known administration routes can be used, and oral or parenteral administration can be used. Because of ease of administration, the growth inhibitor is preferably administered parenterally, and more preferably applied externally. When administered orally, the growth inhibitor may be used as a pharmaceutical product or a food or beverage. From the viewpoint of the effect of inhibiting bacterial growth, the growth inhibitor is preferably used as a cleanser or external preparation.

[0051] The target site of the cleanser or topical agent is a site where at least one bacterium selected from the group consisting of the genus Gardnerella and the genus Atopobium is present or suspected to be present, preferably the female or male intimate area, more preferably the female intimate area, and even more preferably the vagina. In this specification, the female intimate area refers to the vulva, vagina, and surrounding skin, and the male intimate area refers to the penis, testicles, and surrounding skin.

[0052] An external preparation means a preparation to be applied directly to the surface of the skin, mucous membrane, etc., and may be any of a pharmaceutical composition, a quasi-drug, or a cosmetic. The external preparation may be in any form, including solid, semi-solid, liquid, emulsion, cream, gel, mousse, spray, or a two-component system using a mixture of these, so long as it can be applied to or penetrated into the skin, mucous membrane, etc., and there are no limitations on its properties or dosage form.

[0053] The cleanser refers to a preparation for cleansing the skin, mucous membranes, etc., and may be any of a pharmaceutical composition, a quasi-drug, or a cosmetic. The cleanser may be in any form, including solid, semi-solid, liquid, emulsion, cream, gel, mousse, spray, or a two-component formulation in which these are mixed, without any limitations on its properties and formulation.

[0054] The cleanser may contain a surfactant and may be a cleanser that is typically foamed before use (e.g., soap, body wash), or may be a cleanser that is filled into a cleaner and sprayed onto the area to be cleaned.

[0055] The wash is preferably a vaginal wash. The vaginal wash is filled, for example, in a container (vaginal wash) equipped with a nozzle that can be inserted into the vagina, and sprayed into the vagina to wash away residual blood and / or secretions remaining inside or outside the vagina.

[0056] <Preparation Containing Antiproliferative Agent> The antiproliferative agent may be administered to a living body as it is, or may be administered as a preparation in which an effective amount of the antiproliferative agent is formulated with a pharmaceutically acceptable carrier.

[0057] The dosage form of the preparation is not particularly limited and may be any of a drug, a quasi-drug, or a cosmetic. In other words, one aspect of the present invention is a cosmetic or a quasi-drug containing the growth inhibitor. Another aspect of the present invention is a pharmaceutical composition containing the growth inhibitor.

[0058] The content of glyceryl glucoside in the formulation can be set appropriately depending on the form and / or application of the formulation, but from the viewpoint of the effect of inhibiting bacterial growth, examples include 0.002 mass% or more, 0.003 mass% or more, 0.004 mass% or more, 0.005 mass% or more, 0.006 mass% or more, 30 mass% or less, 29 mass% or less, 28 mass% or less, 27 mass% or less, 26 mass% or less, 25 mass% or less, 24 mass% or less, 23 mass% or less, 22 mass% or less, 21 mass% or less, and 20 mass% or less.

[0059] The cosmetic or quasi-drug is preferably a cosmetic or quasi-drug for the delicate zone, more preferably a cream, ointment, gel, or lotion for the delicate zone.

[0060] The cosmetic or quasi-drug may contain, as needed, ingredients typically used in cosmetics, pharmaceutical skin care products, external preparations, or cleansers, such as fats and oils, waxes, hydrocarbon oils, ester oils, higher alcohols, silicone oils, moisturizers, surfactants, water-soluble polymers, thickeners, powders, skin protectants, whitening agents, wrinkle treatment agents, anti-aging agents, plant extracts, preservatives, anti-inflammatory agents, pH adjusters, sequestering agents, antioxidants, stabilizers, fragrances, colorants, pigments, etc., within limits that do not impair the effects of the present invention. These ingredients may be used alone or in combination of two or more. The cosmetic or quasi-drug may also contain probiotics, such as Döderlein's bacillus.

[0061] The pharmaceutical composition is preferably for treating or preventing a disease associated with Gardnerella vaginalis or Atopobium vaginae, such as bacterial vaginosis, urinary tract infection, endometritis, acute salpingitis, postoperative infection, amniotic fluid infection, infertility, premature birth, and neonatal sepsis, with bacterial vaginosis being preferred.

[0062] The dosage form of the pharmaceutical composition is not particularly limited, and may be a solid preparation such as a powder, granules, tablets, or capsules; or a liquid preparation such as a solution, suspension, or emulsion.

[0063] The pharmaceutical composition is preferably a pharmaceutical composition for the intimate area, more preferably a vaginal suppository or a cream, ointment, gel, or lotion to be applied to the skin or mucous membrane of the intimate area, and even more preferably a vaginal suppository or a cream, ointment, gel, or lotion to be applied to the skin or mucous membrane of a woman's intimate area. A vaginal suppository is an external preparation to be applied to the vagina, and may be in any dosage form such as a tablet, capsule, ointment, or gel, and may be inserted into the vagina using an applicator.

[0064] The pharmaceutical composition may contain antibiotics, antifungals, germicides, disinfectants, local anesthetics, antihistamines, antipruritics, preservatives, pH adjusters, excipients, binders, disintegrants, lubricants, stabilizers, solubilizers, suspending agents, isotonicity agents, buffers, antioxidants, probiotics such as Doderlein's bacillus, etc. These components may be used alone or in combination of two or more.

[0065] Antibiotics include, for example, metronidazole, clindamycin, secnidazole, chloramphenicol, and nifrateral.

[0066] Antifungal agents include, for example, fluconazole, butoconazole, clotrimazole, miconazole, and tioconazole.

[0067] Examples of disinfectants include benzalkonium chloride, benzethonium chloride, propyl iodine, and isopropyl methylphenol.

[0068] Examples of local anesthetics include lidocaine, xylocaine, dibucaine, and ethyl aminobenzoate.

[0069] Antihistamines include, for example, diphenhydramine, cetirizine, doxepin, and hydroxyzine.

[0070] An example of an antipruritic agent is crotamiton.

[0071] The preparation can be produced by adding the antiproliferative agent according to a method commonly used for these preparations. The antiproliferative agent may be added at the beginning, middle or end of the production process of the preparation, and the method of addition may be selected from mixing, kneading, dissolving, immersing, sprinkling, spraying, coating, etc., depending on the form of the preparation.

[0072] <Method for inhibiting bacterial growth> One aspect of the present invention is a method for inhibiting the growth of at least one bacterium selected from the group consisting of the genus Gardnerella and the genus Atopobium, which comprises the step of administering glyceryl glucoside to a subject.

[0073] The method of administering glyceryl glucoside to a subject is not particularly limited, and any suitable means can be used depending on the subject. As the administration method, parenteral administration is preferred, and external application to the skin or mucosa is more preferred. External application to the skin or mucosa may be, for example, a simple application method, layering therapy, occlusive therapy, moist therapy, etc.

[0074] The target site to which glyceryl glucoside is administered is a site where at least one bacterium selected from the group consisting of the genus Gardnerella and the genus Atopobium is present or suspected to be present, preferably the intimate area of ​​a woman or a man, more preferably the intimate area of ​​a woman, and even more preferably the vagina.

[0075] One aspect of the present invention is a therapeutic or preventive agent for a disease associated with Gardnerella vaginalis or Atopobium vaginae, which comprises glyceryl glucoside.

[0076] One aspect of the present invention is glyceryl glucoside for use as a therapeutic or preventative agent for diseases involving Gardnerella vaginalis or Atopobium vaginae.

[0077] One aspect of the present invention is the use of glyceryl glucoside to inhibit the growth of at least one bacterium selected from the group consisting of the genera Gardnerella and Atopobium.

[0078] One aspect of the present invention is the use of glyceryl glucoside in the manufacture of an inhibitor of the growth of at least one bacterium selected from the group consisting of the genera Gardnerella and Atopobium.

[0079] One aspect of the present invention is a method for treating or preventing a disease associated with Gardnerella vaginalis or Atopobium vaginae, comprising administering an effective amount of glyceryl glucoside to a subject.

[0080] The present invention will now be described in more detail with reference to examples, but the present invention is not limited to these examples.

[0081] Example 1 Growth Test Using Gardnerella vaginalis, Atopobium vaginae, and Döderlein's Bacillus (Test Substance) C-mo 2G (manufactured by Toyo Sugar Refining Co., Ltd.) was used as the glyceryl glucoside. Penicillin-streptomycin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd., No. 168-23191) was used as a positive control. Sterilized water was used as a control.

[0082] (Bacteria and Culture Media) The bacteria and culture media shown in Table 1 were used and cultured under the conditions shown in Table 1.

[0083]

[0084] The GAM semi-solid high layer medium used was No. 05424 manufactured by Nissui Pharmaceutical Co., Ltd. The number of days for culture was set to a number of days until bacterial growth could be visually confirmed in the control.

[0085] Although various Döderlein's bacilli are known, experiments were conducted using Lactobacillus crispatus and Lactobacillus iners as representative Döderlein's bacilli because they are the predominant bacteria in the vagina in all four ethnic groups (Caucasians, Blacks, Hispanics, and Asians). Experiments were also conducted using Atopobium vaginae as a representative bacterium of the Atopobium genus.

[0086] (Preculture) Each bacterium, which had been frozen and stocked (medium containing 15% (vol / vol) glycerin), was inoculated into 9 mL of medium and culture was initiated. After culturing until the logarithmic growth phase, the medium was thoroughly stirred to homogenize the turbidity, and the absorbance at 600 nm was measured. The medium was diluted to an absorbance (600 nm) of 0.1, and this was used as a bacterial dilution.

[0087] (Test method) C-mo 2G was added to the medium to a final concentration of 0.5%, 1.0%, or 2.0%. As a positive control, penicillin-streptomycin was added to Lactobacillus crispatus at a final concentration of 10%, Lactobacillus iners at a final concentration of 1%, Gardnerella vaginalis at a final concentration of 1%, and Atopobium vaginae at a final concentration of 10%.

[0088] 200 μL of the bacterial dilution was added to 9 mL of medium containing the test substance, and cultivation was initiated. After cultivation, the medium was thoroughly stirred to make it uniformly turbid, and then the absorbance (turbidity) at 600 nm was measured. The test was performed with n=3 for each point. A significant difference test was performed using Student's t-test, with *: p<0.05 and **: p<0.01 compared to the control.

[0089] (Results) The growth test results for each bacterium are shown in Figures 1 and 2. In Figure 1, the vertical axis shows the measured values ​​of absorbance (600 nm) (average value of n = 3). In Figure 2, the vertical axis shows the relative growth rate when the average value of absorbance (600 nm) of the control is set to 100%, and shows the relative value of bacterial growth to the control.

[0090] In the positive control, the growth of all bacteria, Lactobacillus crispatus, Lactobacillus iners, Gardnerella vaginalis, and Atopobium vaginae, was strongly inhibited. The addition of glyceryl glucoside significantly suppressed the growth of Gardnerella vaginalis and Atopobium vaginae, but the number of Lactobacillus crispatus remained unchanged, while the number of Lactobacillus iners increased compared to the control. Furthermore, the addition of glyceryl glucoside significantly suppressed the growth of Gardnerella vaginalis and Atopobium vaginae, and this growth-inhibitory effect was concentration-dependent.

[0091] That is, it was revealed that glyceryl glucoside did not inhibit the growth of Lactobacillus crispatus and Lactobacillus iners, which are Doderlein's bacilli, but inhibited the growth of Gardnerella vaginalis and Atopobium vaginae.

[0092] Reference Example 1 Growth Test Using Propionibacterium acnes, Streptococcus mutans, and Staphylococcus aureus (Test Substance) COSARTE-2G (manufactured by Toyo Sugar Refining Co., Ltd.) was used as glyceryl glucoside, and was diluted with ion-exchanged water to a concentration of 40% by mass. BD (Sensi-Disc Tetracyclone 30 Drug Sensitivity Kit, manufactured by Nippon Pepton Tickison Co., Ltd.) was used as a positive control. Glycerin (manufactured by Nacalai Tesque, Inc.) was used as a control, and was diluted with ion-exchanged water to a concentration of 40% by mass.

[0093] (Propionibacterium acnes) As a representative strain of Propionibacterium acnes, Propionibacterium acnes (JCM 6425) was used. The Propionibacterium acnes strain was placed in a GAM liquid medium, which was then placed in an Anaeropack at 35°C and pre-cultured under anaerobic conditions.

[0094] (Dental caries bacteria) Streptococcus mutans (JCM 5705) was used as a representative strain of dental caries bacteria. The dental caries bacteria strain was placed in a BHI liquid medium and pre-cultured at 35°C.

[0095] (Staphylococcus aureus) Staphylococcus aureus (NBRC 13276) was used as a representative strain of Staphylococcus aureus. The Staphylococcus aureus strain was placed in a BHI liquid medium and pre-cultured at 37°C.

[0096] (Test Method) GAM agar plate medium and BHI agar plate medium were prepared. Pre-cultured Propionibacterium acnes strain, Propionibacterium dental caries strain, and Staphylococcus aureus strain in the logarithmic growth phase were added at 1 × 10 7The test substance was suspended in liquid medium to a concentration of 1000 cells / mL, and 50 μL of this suspension was smeared onto each agar plate using a Conlarger rod. Each test substance was sterilized by filtration using a membrane filter, and then 50 μL of the substance was impregnated into a sterilized paper disk. The paper disk was placed aseptically on the agar plate and cultured for 48 hours under conditions suitable for each bacterial strain. The bacterial growth inhibitory effect was assessed based on the presence and size of a growth inhibition zone around the paper disk.

[0097] (Results) In BD (positive control), large zones of growth inhibition were observed for all of Propionibacterium acnes, Streptococcus mutans, and Staphylococcus aureus, and the growth of these bacteria was strongly suppressed.

[0098] Glyceryl glucoside did not show any growth inhibition zones for Propionibacterium acnes, Streptococcus mutans, or Staphylococcus aureus. This indicates that glyceryl glucoside does not inhibit the growth of Propionibacterium acnes, Streptococcus mutans, or Staphylococcus aureus. The results for glycerin, used as a control, were similar to those for glyceryl glucoside.

Claims

1. A growth inhibitor for at least one bacterium selected from the group consisting of Gardnerella and Atopobium, comprising glyceryl glucoside.

2. The proliferation inhibitor according to claim 1, wherein the bacterium is at least one selected from the group consisting of Gardnerella vaginalis and Atopobium vaginae.

3. The proliferation inhibitor according to claim 1 or 2, which is used as a cleansing agent or an external preparation.

4. A cosmetic or quasi-drug comprising the proliferation inhibitor according to claim 1 or 2.

5. A pharmaceutical composition for treating or preventing a disease involving Gardnerella vaginalis or Atopobium vaginae, comprising the proliferation inhibitor according to claim 2.

6. The pharmaceutical composition according to claim 5, wherein the disease is bacterial vaginosis.

7. A method for inhibiting the growth of at least one bacterium selected from the group consisting of the genera Gardnerella and Atopobium, which comprises a step of administering glyceryl glucoside to a subject.

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