Autoinducer-2 inhibitor

Abstract

To provide an AI-2 inhibitor having the activity of inhibiting autoinducer-2.SOLUTION: When copper chlorophyllin alkali metal salt, vitamin B2, tin fluoride, zinc chloride, and aluminium lactate are used in a concentration range in which they do not exhibit antibacterial activity (i.e, a concentration range in which they do not inhibit the proliferation and / or growth of bacteria), they show excellent activity of inhibiting AI-2 and can be used as active ingredients of an AI-2 inhibitor.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to an autoinducer-2 inhibitor that has inhibitory activity against autoinducer-2 and can inhibit bacterial quorum sensing. [Background technology]

[0002] Quorum sensing is a microbial signaling mechanism that senses the density of bacteria by sensing the extracellular concentration of autoinducers, which are signal substances produced by the same bacteria, and controls specific gene expression and phenotype accordingly. Quorum sensing is known to be involved in bacterial infection, pathogenicity expression, biofilm formation, toxin production, etc.

[0003] For example, many pathogenic bacteria such as Streptococcus mutans, which causes dental caries, Porphyromonas gingivalis, which causes periodontal disease, Helicobacter pylori, which causes gastric ulcers and gastric cancer, and Clostridium perfringens, which causes food poisoning and hemorrhagic enteritis, use autoinducer-2 (AI-2) as a signal substance to control quorum sensing. Therefore, by using a component that has inhibitory activity against AI-2, it is possible to inhibit quorum sensing of these pathogenic bacteria, and as a result, it is expected that various infectious diseases caused by these pathogenic bacteria can be prevented or treated.

[0004] Various components that have inhibitory activity against AI-2 have been studied. For example, Patent Document 1 lists 5-amino-n-valeric acid, butane-1,4-diamine, guanine, acetoacetic acid, γ-hydroxybutyric acid, piperidine, gluconic acid, 3-hydroxy-3-methylglutaric acid, N 8 It is disclosed that 5-acetylspermidine and urocanic acid, as well as salts thereof, can be used as AI-2 inhibitors. In addition, Patent Document 2 discloses that 4,5-dihydroxy-2-cyclopenten-1-one can be used as an AI-2 inhibitor.

[0005] However, in order to further improve the inhibitory activity of AI-2 and to keep up with the diversification of products that have inhibitory activity against AI-2, there is a strong demand for the development of new components that have inhibitory activity against AI-2. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] JP 2012-97036 A [Patent Document 2] WO 2003 / 77844 Summary of the Invention [Problem to be solved by the invention]

[0007] An object of the present invention is to provide an AI-2 inhibitor having inhibitory activity against AI-2. [Means for solving the problem]

[0008] The present inventors have conducted extensive research to solve the above problems and have found that copper chlorophyllin alkali metal salt, vitamin B2, tin fluoride, zinc chloride, and aluminum lactate exhibit excellent inhibitory activity against AI-2 when used in a concentration range that does not exert antibacterial activity (i.e., a concentration range that does not suppress bacterial proliferation and / or growth), and can be used as active ingredients of AI-2 inhibitors. The present invention was completed based on this finding and through further research.

[0009] That is, the present invention provides the following aspects. Item 1. An AI-2 inhibitor containing as an active ingredient at least one selected from the group consisting of copper chlorophyllin alkali metal salt, vitamin B2, tin fluoride, zinc chloride, and aluminum lactate. Item 2. The active ingredient is copper chlorophyllin alkali metal salt, Item 2. The AI-2 inhibitor according to Item 1, wherein the concentration of the copper chlorophyllin alkali metal salt is 10 to 2500 μM at the site where AI-2 inhibition is required. Item 3. The active ingredient is vitamin B2, Item 2. The AI-2 inhibitor according to Item 1, which is used so that the concentration of vitamin B2 at the site where AI-2 inhibition is required is 20 to 800 μM. Item 4. The active ingredient is stannous fluoride, Item 2. The AI-2 inhibitor according to item 1, wherein the stannous fluoride is used at a concentration of 10 to 4500 μM at the site where inhibition of AI-2 is required. Item 5. The active ingredient is zinc chloride, Item 2. The AI-2 inhibitor according to Item 1, which is used so that the concentration of zinc chloride at the site where AI-2 inhibition is required is 20 to 2000 μM. Item 6. The active ingredient is aluminum lactate, Item 2. The AI-2 inhibitor according to Item 1, which is used so that the concentration of aluminum lactate at the site where AI-2 inhibition is required is 100 to 3500 μM. Item 7. The AI-2 inhibitor according to any one of Items 1 to 6, which is an oral care product. Effect of the Invention

[0010] According to the present invention, AI-2 can be inhibited without exerting antibacterial activity, and therefore quorum sensing of pathogenic bacteria can be inhibited without adversely affecting resident bacteria, making it possible to prevent or treat infectious diseases. [Brief description of the drawings]

[0011] [Figure 1] In Test Example 1, Vibrio harveyi MM32 strain was cultured in the presence of various concentrations of sodium copper chlorophyllin, and the AI-2 inhibitory activity and viable cell count were measured. [Diagram 2] In Test Example 2, Vibrio harveyi MM32 strain was cultured in the presence of various concentrations of vitamin B2, and the AI-2 inhibitory activity and viable cell count were measured. [Diagram 3]In Test Example 3, Vibrio harveyi MM32 strain was cultured in the presence of various concentrations of stannous fluoride, and the AI-2 inhibitory activity and viable cell count were measured. [Figure 4] In Test Example 4, Vibrio harveyi MM32 strain was cultured in the presence of zinc chloride at various concentrations, and the AI-2 inhibitory activity and viable cell count were measured. [Diagram 5] In Test Example 5, Vibrio harveyi MM32 strain was cultured in the presence of aluminum lactate at various concentrations, and the AI-2 inhibitory activity and viable cell count were measured. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] The AI-2 inhibitor of the present invention is characterized in that it contains at least one active ingredient selected from the group consisting of copper chlorophyllin alkali metal salt, vitamin B2, tin fluoride, zinc chloride, and aluminum lactate. The AI-2 inhibitor of the present invention will be described in detail below.

[0013] [Active ingredients] The AI-2 inhibitor of the present invention uses copper chlorophyllin alkali metal salt, vitamin B2, tin fluoride, zinc chloride, and / or aluminum lactate as active ingredients. When these active ingredients are allowed to coexist with bacteria that control quorum sensing using AI-2 as a signal substance at a concentration that does not exert the antibacterial activity of these active ingredients (i.e., under conditions in which the bacteria are alive), AI-2 in the bacteria can be inhibited.

[0014] Among the above-mentioned effective ingredients, the copper chlorophyllin alkali metal salt specifically includes sodium copper chlorophyllin and potassium copper chlorophyllin. The copper chlorophyllin alkali metal salt is preferably sodium copper chlorophyllin.

[0015] Among the above-mentioned active ingredients, the tin fluoride may be either tin (II) fluoride or tin (IV) fluoride, with tin (II) fluoride being preferred.

[0016] Among the above-mentioned active ingredients, specific examples of vitamin B2 include riboflavin, riboflavin butyrate, riboflavin phosphate, and salts of riboflavin phosphate.Specific examples of salts of riboflavin phosphate include alkali metal salts such as sodium salt and potassium salt.Preferably, vitamin B2 includes riboflavin phosphate and its salts, and more preferably, sodium riboflavin phosphate.

[0017] In the AI-2 inhibitor of the present invention, the active ingredient may be one selected from copper chlorophyllin alkali metal salt, vitamin B2, tin fluoride, zinc chloride, and aluminum lactate and used alone, or two or more of these may be used in combination.

[0018] [Concentration used] The AI-2 inhibitor of the present invention is used at a concentration where the active ingredient does not exert its antibacterial activity (i.e., under conditions where bacteria are alive). By using the active ingredient at a concentration where the antibacterial activity is not exerted in this way, it is possible to prevent or treat infectious diseases caused by bacteria that control quorum sensing using AI-2 as a signal substance, while avoiding the active ingredient from adversely affecting normal bacteria.

[0019] In the AI-2 inhibitor of the present invention, the concentration of the active ingredient during use may be within a range in which the active ingredient does not exert its antibacterial activity and can exert its inhibitory activity against AI-2, and may be appropriately set depending on the type of active ingredient used. Specific examples of the concentration of each active ingredient during use include the following ranges: Copper chlorophyllin alkali metal salt : 10 to 2500 μM, preferably 50 to 2500 μM, more preferably 100 to 2500 μM, and even more preferably 500 to 2000 μM. Vitamin B 2 in the case of : 20 to 800 μM, preferably 50 to 800 μM, and more preferably 100 to 200 μM. In the case of tin fluoride : 10 to 4500 μM, preferably 50 to 4500 μM, more preferably 100 to 4500 μM. In the case of zinc chloride : 20 to 2000 μM, preferably 50 to 2000 μM. Aluminum Lactate : 100 to 3500 μM, preferably 1500 to 3500 μM, more preferably 2000 to 3500 μM, and even more preferably 2500 to 3500 μM.

[0020] Here, the "concentration of the active ingredient during use" refers to the concentration at which the active ingredient acts at the site where inhibition of AI-2 is required, i.e., the concentration of the active ingredient when coexisting with bacteria that control quorum sensing using AI-2 as a signal substance. For example, when the AI-2 inhibitor of the present invention is used as an oral care product, the active ingredient diluted by saliva or the like in the oral cavity may satisfy the above-mentioned concentration range.

[0021] [Application] The AI-2 inhibitor of the present invention is used to inhibit AI-2 in bacteria that control quorum sensing using AI-2 as a signal substance. By inhibiting AI-2 in the bacteria, the quorum sensing can be inhibited, and as a result, pathogenic expression, biofilm formation, toxin production, etc. can be suppressed, and an effect of preventing or treating infections caused by the bacteria can be achieved. Therefore, the AI-2 inhibitor of the present invention can be used as an agent for preventing or treating infections caused by bacteria that control quorum sensing using AI-2 as a signal substance.

[0022] Examples of bacteria that control quorum sensing using AI-2 as a signal substance include Streptococcus mutans, Streptococcus pneumoniae, Streptococcus gordonii, Streptococcus oralis, Streptococcus sobrinus, and Streptococcus anginosus; Porphyromonas gingivalis; Clostridium perfringens and Clostridium difficile; Helicobacter pylori; Vibrio harveyi, Vibrio cholerae, and Vibrio fischeri; and Enterobacter cloacae. Examples of bacteria that can be used include Enterobacter bacteria; Staphylococcus bacteria such as Staphylococcus aureus; Pseudomonas bacteria such as Pseudomonas aeruginosa; Salmonella bacteria such as Salmonella enterica and Salmonella typhimurium; Campylobacter bacteria such as Campylobacter jejuni and Campylobacter coli; Escherichia bacteria such as Escherichia coli; Actinomyces bacteria such as Actinomyces naeslundii; Fusobacterium bacteria such as Fusobacterium nucleatum; Aggregatibacter bacteria such as Aggregatibacter actinomycetemcomitans; and Prevotella bacteria such as Prevotella intermedia.

[0023] For example, by inhibiting AI-2 of Streptococcus mutans, its quorum sensing can be inhibited, and as a result, the formation of biofilms that cause dental caries and the acquisition of acid resistance by the bacteria can be suppressed. Therefore, the AI-2 inhibitor of the present invention can be used as an agent for preventing or treating dental caries.

[0024] By inhibiting AI-2 of Porphyromonas gingivalis, Streptococcus gordonii, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, or Prevotella intermedii, it is possible to inhibit their quorum sensing, thereby suppressing the formation of biofilms that cause periodontal disease and the production of toxins by the bacteria, etc., and therefore the AI-2 inhibitor of the present invention can be used as an agent for preventing or treating periodontal disease.

[0025] By inhibiting AI-2 of Actinomyces naeslundii, its quorum sensing can be inhibited, and as a result, the formation of biofilms that cause dental caries or periodontal disease can be suppressed. Therefore, the AI-2 inhibitor of the present invention can be used as an agent for preventing dental caries or periodontal disease.

[0026] By inhibiting AI-2 of food poisoning bacteria such as Salmonella enterica, Salmonella typhimurium, Campylobacter jejuni, Campylobacter coli, and Escherichia coli, it is possible to inhibit their quorum sensing, thereby suppressing the production of toxins that cause food poisoning. Therefore, the AI-2 inhibitor of the present invention can be used as an agent for preventing or treating food poisoning.

[0027] By inhibiting AI-2 of Clostridium perfringens, the production of enterotoxins in the intestines can be suppressed, and therefore the AI-2 inhibitor of the present invention can be used as an agent for preventing or treating food poisoning, hemorrhagic enteritis, or gas gangrene.

[0028] By inhibiting AI-2 of Helicobacter pylori, the production of urease can be inhibited and the establishment of the bacterium in the stomach can be suppressed, so the AI-2 inhibitor of the present invention can be used as an agent for preventing or treating gastritis, gastric ulcers or gastric cancer.

[0029] By inhibiting AI-2 of Enterobacter bacteria, the production of Shiga toxin by enterohemorrhagic Escherichia coli can be suppressed, and therefore the AI-2 inhibitor of the present invention can be used as an agent for preventing or treating enterohemorrhagic Escherichia coli infections.

[0030] By inhibiting AI-2 of Staphylococcus aureus, the production of toxins by Staphylococcus aureus can be suppressed, and therefore the AI-2 inhibitor of the present invention can be used as an agent for preventing or treating atopic dermatitis.

[0031] Since the toxin production by Pseudomonas aeruginosa can be suppressed by inhibiting AI-2 of Pseudomonas aeruginosa, the AI-2 inhibitor of the present invention can be used as an agent for preventing or treating Pseudomonas aeruginosa infections.

[0032] [Products containing AI-2 inhibitors] The dosage form of the product containing the AI-2 inhibitor of the present invention is not particularly limited, and may be any of solid, semi-solid, and liquid.

[0033] Specific examples of products in which the AI-2 inhibitor of the present invention is incorporated include oral care products, pharmaceuticals, foods and beverages, etc. Among these, oral care products are preferred.

[0034] When the AI-2 inhibitor of the present invention is incorporated into an oral care product (i.e., when it is provided as an oral care product for AI-2 inhibition), the active ingredient may be prepared into a desired form as is or in combination with other additive ingredients. The oral care product may be any product that can be applied to the oral cavity and remain there for a certain period of time, and specific examples of the oral care product include oral hygiene products such as liquid dentifrices, toothpastes, mouthwashes, mouth fresheners (mouth sprays, etc.), oral pastes, and gum massage creams.

[0035] When the AI-2 inhibitor of the present invention is incorporated into an oral care product, the content of the active ingredient in the oral care product may be appropriately set depending on the type of oral care product, the type of active ingredient, etc., so that the active ingredient in the oral cavity when applied to the oral cavity is within the concentration range during use described above. For example, the content of each active ingredient in the oral care product may be in the following ranges. Copper chlorophyllin alkali metal salt : 0.0007 to 0.18% by weight, preferably 0.036 to 0.18% by weight, and more preferably 0.036 to 0.15% by weight. Vitamin B 2 in the case of : 0.0009 to 0.038% by weight, preferably 0.0047 to 0.038% by weight, and more preferably 0.0047 to 0.01% by weight. In the case of tin fluoride : 0.0002 to 0.4% by weight, preferably 0.0016 to 0.4% by weight, and more preferably 0.0016 to 0.07% by weight. In the case of zinc chloride : 0.0003 to 0.1% by weight, preferably 0.0007 to 0.1% by weight, and more preferably 0.0007 to 0.03% by weight. Aluminum Lactate : 0.002 to 1.0% by weight, preferably 0.07 to 1.0% by weight, and more preferably 0.07 to 0.1% by weight.

[0036] When the AI-2 inhibitor of the present invention is incorporated into a medicine (including quasi-drugs) (i.e., when provided as a medicine for AI-2 inhibition), the active ingredient may be prepared into a desired form as is or in combination with other additives, etc. Specific examples of the medicine include internal medicines such as tablets, pills, powders, fine granules, granules, capsules (including hard capsules and soft capsules), troches, chewables, extracts (including soft extracts, dry extracts, etc.), jellies, syrups, spirits, elixirs, and liposome preparations; external medicines such as ointments, creams, lotions, gels, sprays, patches, emulsions, suspensions, poultices, liniments, aerosols, ointments, packs, inhalants, and suppositories; and injections.

[0037] When the AI-2 inhibitor of the present invention is incorporated into a pharmaceutical, the content of the active ingredient in the pharmaceutical may be appropriately set depending on the type of pharmaceutical, the type of active ingredient, etc., so that the active ingredient at the affected area after administration is within the concentration range at the time of use described above. For example, the content of each active ingredient in the pharmaceutical may be in the following ranges. Copper chlorophyllin alkali metal salt : 0.0007 to 0.18% by weight, preferably 0.036 to 0.18% by weight, and more preferably 0.036 to 0.15% by weight. Vitamin B 2 in the case of : 0.0009 to 0.038% by weight, preferably 0.0047 to 0. 0.038% by weight, more preferably 0.0047 to 0.01% by weight. In the case of tin fluoride : 0.0002 to 0.4% by weight, preferably 0.0016 to 0.4% by weight, and more preferably 0.0016 to 0.07% by weight. In the case of zinc chloride : 0.0003 to 0.1% by weight, preferably 0.0007 to 0.1% by weight, and more preferably 0.0007 to 0.03% by weight. Aluminum Lactate : 0.002 to 1.0% by weight, preferably 0.07 to 1.0% by weight, and more preferably 0.07 to 0.1% by weight.

[0038] When the AI-2 inhibitor of the present invention is incorporated into a food or drink (i.e., when it is provided as a food or drink for AI-2 inhibition), the active ingredient may be prepared into a desired form as it is or in combination with other food materials or additives. Examples of the food or drink include general food and drink, as well as foods with health claims (including foods for specified health uses, foods with nutritional functions, and foods with functional claims), foods for sick people, and the like. The form of these foods and drinks is not particularly limited, but specifically includes beverages such as tea drinks, nutritional drinks, fruit juice drinks, carbonated drinks, and lactic acid drinks; supplements such as capsules (soft capsules, hard capsules), tablets, granules, powders, jellies, and liposome preparations; and luxury items such as gummies, candies, and jellies. Among these foods and drinks, supplements are preferred.

[0039] When the AI-2 inhibitor of the present invention is incorporated into food or beverages, the content of the active ingredient in the food or beverage may be appropriately set depending on the type of food or beverage, the type of active ingredient, etc., so that the concentration of the active ingredient in the part of the body where AI-2 inhibition is required after ingestion falls within the concentration range during use described above. For example, the content of each active ingredient in the food or beverage may be in the following ranges. Copper chlorophyllin alkali metal salt : 0.0007 to 0.18% by weight, preferably 0.036 to 0.18% by weight, and more preferably 0.036 to 0.15% by weight. Vitamin B 2 in the case of : 0.0009 to 0.038% by weight, preferably 0.0047 to 0. 0.038% by weight, more preferably 0.0047 to 0.01% by weight. In the case of tin fluoride : 0.0002 to 0.4% by weight, preferably 0.0016 to 0.4% by weight, and more preferably 0.0016 to 0.07% by weight. In the case of zinc chloride : 0.0003 to 0.1% by weight, preferably 0.0007 to 0.1% by weight, and more preferably 0.0007 to 0.03% by weight. Aluminum Lactate : 0.002 to 1.0% by weight, preferably 0.07 to 1.0% by weight, and more preferably 0.07 to 0.1% by weight. EXAMPLES

[0040] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples.

[0041] Test Example 1: Verification of AI-2 inhibitory activity of sodium copper chlorophyllin 1. Test Method The AI-2 inhibitory activity was measured using reporter bacteria that emit light upon recognition of AI-2. The test materials and measurement conditions used are as follows.

[0042] (1) Test materials Reporter bacteria Vibrio harveyi MM32 strain (ATCC BAA-1121) was used as the reporter bacterium.

[0043] ·Km-LB medium LB medium containing 10 g / L sodium chloride, 10 g / L bactotryptone, and 5 g / L yeast extract was sterilized by high-pressure steam (121°C, 15 minutes), and then kanamycin sulfate was added aseptically to a concentration of 50 mg / L to prepare Km-LB medium.

[0044] AB medium First, an aqueous solution containing 0.3 M sodium chloride, 0.05 M magnesium sulfate, and 2 g / L casamino acids (vitamin-free, Difco) was adjusted to pH 7.5 with potassium hydroxide, and sterilized by high-pressure steam (121 ° C, 15 minutes). 1 mL of 1 M potassium phosphate buffer (a buffer adjusted to pH 7.0 by mixing 1 M potassium dihydrogen phosphate aqueous solution and 1 M dipotassium hydrogen phosphate aqueous solution), 1 mL of an aqueous solution containing 0.1 M L-arginine, 2 mL of an aqueous solution containing 50 wt% glycerol, 0.01 mL of an aqueous solution containing 0.1 mg / mL riboflavin, and 0.01 mL of an aqueous solution containing 10 mg / mL thiamine hydrochloride were added to 100 mL of this aqueous solution, and filtration sterilization was performed with a filter with a pore size of 0.22 μm to obtain AB medium.

[0045] ·HMF-containing liquid 2.35 mg of 4-hydroxy-5-methyl-3(2H)-furanone (HMF), 1 mL of ethanol, and 9 mL of sterile distilled water were mixed in a glass container to prepare an HMF-containing solution (containing 2,000 μM HMF). HMF is known to have AI-2 activity, and in this test, HMF was used as an AI-2 substitute.

[0046] ·BMF-containing liquid 2.15 mg of 4-bromo-5-(4-methoxyphenyl)-2(5H)-furanone (BMF), 4 mL of ethanol, and 36 mL of sterile distilled water were mixed to prepare a BMF-containing solution (containing 200 μM BMF). BMF is known to have the activity of inhibiting AI-2, and in this test, BMF was used as a positive control.

[0047] Test solution Sodium copper chlorophyllin was added to an aqueous solution containing 10% by volume of ethanol at concentrations of 200 μM, 400 μM, 1000 μM, 1400 μM, 2000 μM, 3000 μM, 4000 μM, 10000 μM, 16000 μM, 20000 μM, 30000 μM, 40000 μM, and 50000 μM to prepare test solutions of various concentrations.

[0048] (2) Measurement conditions The reporter bacteria were precultured in Km-LB medium, and the optical density (OD) of the preculture solution at 600 nm was measured. 600 ) and add an appropriate amount of Km-LB medium to obtain the OD 600 The reporter bacteria solution was adjusted to 0.50, and the reporter bacteria solution was obtained. This reporter bacteria solution was diluted 2,500-fold with AB medium to obtain a reporter bacteria dilution solution.

[0049] In a sterile glass test tube, 0.9 mL of the reporter bacteria dilution solution and 0.05 mL of the test solution were mixed and incubated for 10 minutes under conditions of 30°C and stirring (180 rpm) in the dark. Then, 0.05 mL of HMF-containing solution was added and incubated for 4 hours under conditions of 30°C and stirring (180 rpm) in the dark. At this point, the test solution was diluted 20 times, so the final concentrations of copper chlorophyllin sodium in the solution were 10 μM, 20 μM, 50 μM, 70 μM, 100 μM, 150 μM, 200 μM, 500 μM, 800 μM, 1000 μM, 1500 μM, 2000 μM, and 2500 μM. 0.1 mL of the incubation solution was transferred to a plate for luminescence measurement, and the luminescence intensity at 578 nm (luminescence intensity of the sample) was measured using a microplate reader (instrument: SpextraMax iD3, measurement mode: LUM endpoint, MOLECULAR DEVICE Co., Ltd.). In addition, the number of live reporter bacteria in the incubation solution was measured by colony counting method.

[0050] As a blank, an aqueous solution containing 10% by volume of ethanol was used instead of the test solution, and an aqueous solution containing 10% by volume of ethanol was used instead of the HMF-containing solution, and the luminescence intensity (blank luminescence intensity) and viable cell count were measured under the same conditions as above.

[0051] As a negative control, an aqueous solution containing 10% by volume of ethanol was used instead of the test solution, and the luminescence intensity (luminescence intensity of the negative control) and the viable cell count were measured under the same conditions as above.

[0052] As a positive control, a BMF-containing solution was used instead of the test solution, and the luminescence intensity (luminescence intensity of the positive control) and viable cell count were measured under the same conditions as above.

[0053] The AI-2 inhibition rate was calculated from the measured luminescence intensity according to the following formula.

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[0054] The results are shown in Figure 1. As a result, it was revealed that sodium copper chlorophyllin does not exhibit antibacterial activity in the concentration range of 10 to 2500 μM, and has the activity of effectively inhibiting AI-2. In particular, it was confirmed that sodium copper chlorophyllin exhibits significantly superior AI-2 inhibitory activity in the concentration range of 100 to 2500 μM.

[0055] Test Example 2: Vitamin B 2 Verification of AI-2 inhibitory activity of Vitamin B2 (riboflavin sodium phosphate) was added to an aqueous solution containing 10% by volume of ethanol to give concentrations of 400μM, 1000μM, 2000μM, 3000μM, 4000μM, 10000μM, and 16000μM to prepare test solutions of various concentrations. Using the obtained test solutions, the AI-2 inhibition rate and viable cell count were measured under the same conditions as in Test Example 1. That is, in this test, the final concentrations of vitamin B2 when measuring the AI-2 inhibition rate and viable cell count were 20μM, 50μM, 100μM, 150μM, 200μM, 500μM, and 800μM.

[0056] The results are shown in Figure 2. As a result, it was revealed that vitamin B2 does not exhibit antibacterial activity in the concentration range of 20 to 800 μM, and has the activity of effectively inhibiting AI-2. In particular, it was confirmed that vitamin B2 exhibits significantly superior AI-2 inhibitory activity in the concentration range of 100 to 800 μM.

[0057] Test Example 3: Verification of the AI-2 inhibitory activity of stannous fluoride Tin(II) fluoride was added to an aqueous solution containing 10% by volume of ethanol at 200μM, 1000μM, 2000μM, and 4000μM to prepare test solutions of various concentrations. In addition, bentonite (Benton DE, manufactured by Chelesto Co., Ltd.) was dispersed in sterile distilled water to prepare a bentonite dispersion, and tin(II) fluoride was added to the bentonite dispersion at 10000μM, 20000μM, 40000μM, 70000μM, 80000μM, and 90000μM to prepare test solutions of various concentrations. Using the obtained test solution, the AI-2 inhibition rate and viable cell count were measured under the same conditions as in Test Example 1. That is, in this test, the final concentrations of stannous fluoride at the time of measuring the AI-2 inhibition rate and the viable cell count were 10 μM, 50 μM, 100 μM, 200 μM, 500 μM, 1000 μM, 2000 μM, 3500 μM, 4000 μM, and 4500 μM. Note that in this test, an aqueous solution containing 10% by volume of ethanol or a bentonite dispersion was used for the blank and negative control measurements.

[0058] The results are shown in Figure 3. As a result, it was revealed that tin fluoride does not exhibit antibacterial activity in the concentration range of 10 to 4500 μM, and has the activity of effectively inhibiting AI-2. In particular, it was confirmed that tin fluoride exhibits significantly superior AI-2 inhibitory activity in the concentration range of 100 to 4500 μM.

[0059] Test Example 4: Verification of AI-2 inhibitory activity of zinc chloride Zinc chloride was added to an aqueous solution containing 10% by volume of ethanol so as to have a concentration of 400μM, 1000μM, 4000μM, 10000μM, 16000μM, 20000μM, 30000μM and 40000μM to prepare test solutions of various concentrations. Using the obtained test solutions, the AI-2 inhibition rate and viable cell count were measured under the same conditions as in Test Example 1. That is, in this test, the final concentrations of zinc chloride when measuring the AI-2 inhibition rate and viable cell count were 20μM, 50μM, 200μM, 500μM, 800μM, 1000μM, 1500μM and 2000μM.

[0060] The results are shown in Figure 4. As a result, it was revealed that zinc chloride does not exhibit antibacterial activity in the concentration range of 20 to 2000 μM, and has the activity of effectively inhibiting AI-2. In particular, it was confirmed that zinc chloride exhibits significantly superior AI-2 inhibitory activity in the concentration range of 50 to 2000 μM.

[0061] Test Example 5: Verification of AI-2 inhibitory activity of aluminum lactate Aluminum lactate was added to an aqueous solution containing 10% by volume of ethanol so as to be 2000μM, 3000μM, 4000μM, 6000μM, 10000μM, 20000μM, 30000μM, 40000μM, 50000μM, 60000μM, and 70000μM to prepare test solutions of various concentrations. Using the obtained test solutions, the AI-2 inhibition rate and viable cell count were measured under the same conditions as in Test Example 1. That is, in this test, the final concentrations of aluminum lactate at the time of measuring the AI-2 inhibition rate and viable cell count were 100μM, 150μM, 200μM, 300μM, 500μM, 1000μM, 1500μM, 2000μM, 2500μM, 3000μM, and 3500μM.

[0062] The results are shown in Figure 5. As a result, it was revealed that aluminum lactate does not exhibit antibacterial activity in the concentration range of 100 to 3500 µM, and has the activity of effectively inhibiting AI-2. In particular, it was confirmed that aluminum lactate exhibits significantly superior AI-2 inhibitory activity in the concentration range of 2000 to 3500 µM.

Claims

1. Copper chlorophyllin alkali metal salt, Vitamin B 2 The autoinducer-2 inhibitor contains at least one active ingredient selected from the group consisting of tin fluoride, zinc chloride, and aluminum lactate.

2. The active ingredient is copper chlorophyllin alkali metal salt, The autoinducer-2 inhibitor according to claim 1, wherein the concentration of the copper chlorophyllin alkali metal salt is 10 to 2500 μM at the site where inhibition of autoinducer-2 is required.

3. The active ingredient is vitamin B 2 and Vitamin B12 is required to inhibit autoinducer-2 2 The autoinducer-2 inhibitor according to claim 1, wherein the concentration of the inhibitor is 20 to 800 μM.

4. the active ingredient is stannous fluoride; The autoinducer-2 inhibitor according to claim 1, wherein the concentration of stannous fluoride is 10 to 4500 μM at the site where inhibition of autoinducer-2 is required.

5. The active ingredient is zinc chloride, The autoinducer-2 inhibitor according to claim 1, which is used so that the concentration of zinc chloride at the site where inhibition of autoinducer-2 is required is 20 to 2000 μM.

6. The active ingredient is aluminum lactate, The autoinducer-2 inhibitor according to claim 1, wherein the concentration of aluminum lactate at the site where inhibition of autoinducer-2 is required is 100 to 3500 μM.

7. The autoinducer-2 inhibitor according to any one of claims 1 to 6, which is an oral care product.

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