JP2021183584A - Autoinducer-2 inhibitor - Google Patents

Abstract

[Problem] The object of the present invention is to provide an AI-2 inhibitor having inhibitory activity against autoinducer-2. [Solution] 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 the active ingredient of an AI-2 inhibitor. [Selected Figure] None

Description

The present invention relates to an auto-inducer-2 inhibitor that has inhibitory activity against auto-inducer-2 and can inhibit bacterial quorum sensing.

Quorum sensing is to detect the bacterial density of allogeneic bacteria by sensing the extracellular concentration of auto-inducer, which is a signal substance produced by allogeneic bacteria, and control specific gene expression and phenotype accordingly. It is one of the signal transmission mechanisms between microorganisms. Quorum sensing is known to be involved in bacterial infection, pathogenicity expression, biofilm formation, toxin production and the like.

For example, Streptococcus mutans, the causative agent of caries, Porphyromonas gingivalis, the causative agent of periodontal disease, Helicobacter pylori, which contributes to gastric ulcer and gastric cancer, and contributes to food poisoning and hemorrhagic enteritis. In many pathogenic bacteria such as Clostridium perfringens, autoinducer-2 (hereinafter referred to as AI-2) is used as a signal substance to control quorum sensing. Therefore, by using a component having an inhibitory activity against AI-2, quorum sensing of these pathogenic bacteria can be inhibited, and as a result, various infectious diseases caused by these pathogenic bacteria can be inhibited. Prevention or treatment can be expected.

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

However, in order to further improve the inhibitory activity of AI-2 and follow the diversification of products having the inhibitory activity of AI-2, it is eager to develop a new component having the inhibitory activity against AI-2. Has been done.

Japanese Unexamined Patent Publication No. 2012-97036 International Publication No. 2003/77844

An object of the present invention is to provide an AI-2 inhibitor having an inhibitory activity against AI-2.

As a result of diligent studies to solve the above problems, the present inventor has found that copper chlorophyllin alkali metal salt, vitamin B ₂ , tin fluoride, zinc chloride, and aluminum lactate have a concentration range in which they do not exhibit antibacterial activity (that is,). It has been found that when used in a concentration range that does not suppress the growth and / or growth of bacteria, it exhibits excellent inhibitory activity against AI-2 and can be used as an active ingredient of an AI-2 inhibitor. The present invention has been completed by further studies based on such findings.

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

According to the present invention, since AI-2 can be inhibited without exerting antibacterial activity, quorum sensing of pathogenic bacteria can be inhibited without adversely affecting indigenous bacteria, and infectious diseases can be prevented or treated. Will be possible.

In Test Example 1, the Vibrio Harvey MM32 strain was cultured in the presence of various concentrations of sodium copper chlorophyllin, and the AI-2 inhibitory activity and the viable cell count were measured. In Test Example 2, the _{Vibrio Harvey MM32 strain was cultured in the presence of various concentrations of vitamin B 2} , and the AI-2 inhibitory activity and the viable cell count were measured. In Test Example 3, the Vibrio Harvey MM32 strain was cultured in the presence of various concentrations of tin fluoride, and the AI-2 inhibitory activity and the viable cell count were measured. In Test Example 4, the Vibrio Harvey MM32 strain was cultured in the presence of various concentrations of zinc chloride, and the AI-2 inhibitory activity and the viable cell count were measured. In Test Example 5, the Vibrio Harvey MM32 strain was cultured in the presence of various concentrations of aluminum lactate, and the AI-2 inhibitory activity and the viable cell count were measured.

The AI-2 inhibitor of the present invention _{is characterized by containing at least one selected from the group consisting of copper chlorophyllin alkali metal salt, vitamin B 2} , tin fluoride, zinc chloride, and aluminum lactate as an active ingredient. .. Hereinafter, the AI-2 inhibitor of the present invention will be described in detail.

[Active ingredient]
The AI-2 inhibitor of the present invention uses copper chlorophyllin alkali metal salt, vitamin B ₂ , tin fluoride, zinc chloride, and / or aluminum lactate as active ingredients. When AI-2 is used as a signal substance to coexist with a bacterium controlling quorum sensing at a concentration that does not exert the antibacterial activity of these active ingredients (that is, under the condition that the bacterium is alive), the bacterium is said to be present. Bacterial AI-2 can be inhibited.

Among the active ingredients, examples of the copper chlorophyllin alkali metal salt include copper chlorophyllin sodium and copper chlorophyllin potassium. The copper chlorophyllin alkali metal salt is preferably sodium copper chlorophyllin.

Among the active ingredients, the tin fluoride may be either tin fluoride (II) or tin fluoride (IV), and tin fluoride (II) is preferable.

Among the active ingredients, vitamin B ₂ specifically includes riboflavin, riboflavin butyrate, riboflavin phosphate, salts of riboflavin phosphate and the like. Specific examples of the salt of riboflavin acid include alkali metal salts such as sodium salt and potassium salt. Examples of vitamin B ₂ include riboflavin phosphate and salts thereof, and more preferably riboflavin sodium phosphate.

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

[Concentration used]
The AI-2 inhibitor of the present invention is used at a concentration at which the antibacterial activity of the active ingredient is not exhibited (that is, under conditions in which bacteria are alive). By using the active ingredient at a concentration that does not exert antibacterial activity in this way, while preventing or treating infectious diseases caused by bacteria that control quorum sensing using AI-2 as a signaling substance, the active ingredient is always used. It is possible to prevent the indigenous bacteria from being adversely affected.

In the AI-2 inhibitor of the present invention, the concentration of the active ingredient at the time of use may be as long as the antibacterial activity of the active ingredient is not exhibited and the inhibitory activity against AI-2 can be exhibited. It may be set appropriately according to the type of the component. Specific examples of the concentration of each active ingredient at the time of use include the following ranges.
In the case of copper chlorophyllin alkali metal salt : 10 to 2500 μM, preferably 50 to 2500 μM, more preferably 100 to 2500 μM, still more preferably 500 to 2000 μM.
When Vitamin B _2: 20~800μM, preferably 50～800MyuM, more preferably 100～200MyuM.
For tin fluoride : 10-4500 μM, preferably 50-4500 μM, more preferably 100-4500 μM.
For zinc chloride : 20-2000 μM, preferably 50-2000 μM.
Aluminum lactate : 100 to 3500 μM, preferably 1500 to 3500 μM, more preferably 2000 to 3500 μM, still more preferably 2500 to 3500 μM.

Here, the "concentration of the active ingredient at the time of use" is the concentration at which the active ingredient acts at a site where inhibition of AI-2 is required, that is, quorum sensing is controlled using AI-2 as a signal substance. It is the concentration of the active ingredient when coexisting with the existing bacteria. For example, when the AI-2 inhibitor of the present invention is used as an oral care product, the active ingredient diluted with saliva or the like in the oral cavity may satisfy the above concentration range.

[Use]
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 signaling substance. By inhibiting AI-2 of the bacterium, the quorum sensing can be inhibited, and as a result, pathogenicity expression, biofilm formation, toxin production, etc. can be suppressed, and the preventive or therapeutic effect of the infectious disease caused by the bacterium can be achieved. , The AI-2 inhibitor of the present invention can be used as a prophylactic or therapeutic agent for bacterial infections controlling quorum sensing using AI-2 as a signaling substance.

Examples of the bacterium that controls quorum sensing using AI-2 as a signal substance include Streptococcus mutans, Streptococcus pneumonier, Streptococcus godney, Streptococcus olaris, Streptococcus sobrinus, and Streptococcus anginosus. Bacteria belonging to the genus Porphyromonas such as Porphyromonas gingivalis; Bacteria belonging to the genus Clostridium such as Clostridium perfringens and Clostridium difficile; Vibrio bacterium; Enterobacter bacterium such as Enterobacter cloaka; Staphylococcus bacterium such as Yellow staphylococcus aureus; Pseudomonas bacterium such as Pyogenic bacterium (Pseudomonas erginosa); Salmonella Bacteria of the genus Salmonella such as Enterica, Salmonella tipimurium; Bacteria of the genus Campylobacter such as Campylobacter Jejuni, Campylobacter coli; Bacteria of the genus Escherichia such as Escherichia coli; Examples thereof include bacterium belonging to the genus Fuzobacterium such as Nucleatum; bacterium belonging to the genus Aggregate bacter such as Aggregate bacter actinomycetemcomitans; and bacterium belonging to the genus Prebotera such as Prebotera intermedia.

For example, by inhibiting Streptococcus mutans AI-2, its quorum sensing can be inhibited, and as a result, the formation of a biofilm that causes caries and the acquisition of acid resistance of the bacterium can be suppressed. The AI-2 inhibitor of the present invention can be used as a preventive or therapeutic agent for dental caries.

By inhibiting AI-2 of Porphyromonas gingivalis, Streptococcus godney, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, or Prebotera intermedy, its quorum sensing can be inhibited, resulting in The AI-2 inhibitor of the present invention can be used as a preventive or therapeutic agent for periodontal disease because it can suppress the formation of biofilms that cause periodontal disease and the production of toxins by the bacteria.

By inhibiting Actinomyces Neslandi's AI-2, its quorum sensing can be inhibited, and as a result, biofilm formation that causes caries or periodontal disease can be suppressed. Therefore, the AI- of the present invention can be suppressed. 2 The inhibitor can be used as a preventive agent for dental caries or periodontal disease.

By inhibiting AI-2 of food poisoning bacteria such as Salmonella enterica, Salmonella tipimurium, Campylobacter jejuni, Campylobacter coli, Escherichia coli, etc., its quorum sensing can be inhibited, and as a result, toxins that cause food poisoning. Since the production can be suppressed, the AI-2 inhibitor of the present invention can be used as a preventive or therapeutic agent for food poisoning.

Since the production of enterotoxin in the intestine can be suppressed by inhibiting AI-2 of Clostridium perfringens, the AI-2 inhibitor of the present invention is a preventive or therapeutic agent for food poisoning, hemorrhagic colitis or gas gangrene. Can be used as.

By inhibiting AI-2 of Helicobacter pylori, the production of urease can be inhibited and the colonization of the bacterium in the stomach can be suppressed. Therefore, the AI-2 inhibitor of the present invention can prevent gastritis, gastric ulcer or gastric cancer. It can be used as a therapeutic agent.

Since the production of Shiga toxin in enterohemorrhagic Escherichia coli can be suppressed by inhibiting AI-2 of Enterobacter bacteria, the AI-2 inhibitor of the present invention can be used as a preventive or therapeutic agent for enterohemorrhagic colon infection. Can be used.

Since the production of Staphylococcus aureus toxin can be suppressed by inhibiting AI-2 of Staphylococcus aureus, the AI-2 inhibitor of the present invention can be used as a prophylactic or therapeutic agent for atopic dermatitis.

Since the production of Pseudomonas aeruginosa toxin can be suppressed by inhibiting AI-2 of Pseudomonas aeruginosa, the AI-2 inhibitor of the present invention can be used as a prophylactic or therapeutic agent for Pseudomonas aeruginosa infection. ..

[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 solid, semi-solid, or liquid.

Specific examples of the product containing the AI-2 inhibitor of the present invention include oral care products, pharmaceuticals, foods and drinks, and the like. Among these, oral care products are preferable.

When the AI-2 inhibitor of the present invention is incorporated into an oral care product (that is, when provided as an oral care product for AI-2 inhibition), the active ingredient is desired as it is or in combination with other additive ingredients. It may be prepared in the form of. The oral care product may be any product that can be applied to the oral cavity and stay in the oral cavity for a certain period of time. Specifically, liquid dentifrices, dentifrices, mouthwashes, mouth refreshers (mouth spray, etc.) ), Oral hygiene agents such as oral pasta and gingival massage cream.

When the AI-2 inhibitor of the present invention is blended into an oral care product, the content of the active ingredient in the oral care product is such that the active ingredient in the oral cavity is used as described above when applied into the oral cavity. It may be appropriately set according to the type of oral care product, the type of active ingredient, etc. so as to be within the concentration range at the time. For example, the content of each active ingredient in the oral care product includes the following range. ..
For copper chlorophyllin alkali metal salt : 0.0007 to 0.18% by weight, preferably 0.036 to 0.18% by weight, more preferably 0.036 to 0.15% by weight.
When Vitamin B _2: from .0009 to .038% by weight, preferably 0.0047 to 0.038 wt%, more preferably from 0.0047 to 0.01 wt%.
For tin fluoride : 0.0002 to 0.4% by weight, preferably 0.0016 to 0.4% by weight, 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, 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, more preferably 0.07 to 0.1% by weight.

When the AI-2 inhibitor of the present invention is compounded in a drug (including a quasi-drug) (that is, when it is provided as a drug for AI-2 inhibition), the active ingredient is used as it is or as another additive. It may be prepared in a desired form in combination with the above. Specific examples of pharmaceuticals include tablets, suppositories, powders, fine granules, granules, capsules (including hard capsules and soft capsules), troches, chewables, extracts (soft extracts, dry extracts, etc.). ), Jelly, syrup, liquor, elixir, liposome preparation, etc.; ointments, creams, lotions, gels, sprays, patches, emulsions, suspensions, paps Examples include external pharmaceuticals such as agents, liniment agents, aerosol agents, ointments, packs, inhalants, suppositories; injections and the like.

When the AI-2 inhibitor of the present invention is blended in a drug, the content of the active ingredient in the drug is adjusted so that the active ingredient in the affected area after administration is within the above-mentioned concentration range at the time of use. It may be appropriately set according to the type of the active ingredient, the type of the active ingredient, and the like, and examples thereof include the following ranges as the content of each active ingredient in the pharmaceutical product.
For copper chlorophyllin alkali metal salt : 0.0007 to 0.18% by weight, preferably 0.036 to 0.18% by weight, more preferably 0.036 to 0.15% by weight.
When Vitamin B _2: from 0.0009 to .038% by weight, preferably 0.0047 to 0.038 wt%, more preferably from 0.0047 to 0.01 wt%.
For tin fluoride : 0.0002 to 0.4% by weight, preferably 0.0016 to 0.4% by weight, 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, 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, more preferably 0.07 to 0.1% by weight.

When the AI-2 inhibitor of the present invention is blended into a food or drink (that is, when it is provided as a food or drink for inhibiting AI-2), the active ingredient is desired as it is or in combination with other food materials or additive components. It may be prepared in the form of. Examples of foods and drinks include general foods and drinks, foods with health claims (including foods for specified health use, foods with nutritional claims, and foods with functional claims), foods for the sick, and the like. The form of these foods and drinks is not particularly limited, but specifically, beverages such as tea beverages, nutritional drinks, fruit juice beverages, carbonated beverages, and lactic acid beverages; capsules (soft capsules, hard capsules), tablets, and granules. , Powders, jellies, supplements such as liposome preparations; beverages such as gummy, candy, and jelly. Among these foods and drinks, supplements are preferably mentioned.

When the AI-2 inhibitor of the present invention is blended in a food or drink, the content of the active ingredient in the food or drink is described above as the concentration of the active ingredient in the body part where AI-2 inhibition is required after ingestion. The concentration range at the time of use may be appropriately set according to the type of food and drink, the type of active ingredient, and the like. For example, the content of each active ingredient in food and drink includes the following range.
For copper chlorophyllin alkali metal salt : 0.0007 to 0.18% by weight, preferably 0.036 to 0.18% by weight, more preferably 0.036 to 0.15% by weight.
When Vitamin B _2: .0009 to 0.038 wt%, preferably from 0.0047 to 0.038 wt%, more preferably from 0.0047 to 0.01 wt%.
For tin fluoride : 0.0002 to 0.4% by weight, preferably 0.0016 to 0.4% by weight, 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, 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, more preferably 0.07 to 0.1% by weight.

Hereinafter, the present invention will be specifically described with reference to Examples, but the present invention is not limited to these Examples.

Test Example 1: Verification of AI-2 inhibitory activity of sodium copper chlorophyllin
1. 1. Test method AI-2 inhibitory activity was measured using a reporter bacterium that emits light by recognizing AI-2. The test materials and measurement conditions used are as follows.

(1) Test material / reporter bacterium Vibrio Harvey MM32 strain (ATCC BAA-1121) was used as the reporter bacterium.

-Km-LB medium LB medium containing sodium chloride 10 g / L, bactotrypton 10 g / L, and yeast extract 5 g / L is sterilized by high pressure steam (121 ° C, 15 minutes), and then canamycin sulfate is 50 mg / L. Km-LB medium was prepared by adding it aseptically so as to be.

-AB medium First, an aqueous solution containing 0.3 M sodium chloride, 0.05 M magnesium sulfate, and 2 g / L casamino acid (vitamin-free, manufactured by Difco) was adjusted to pH 7.5 with potassium hydroxide, and the pressure was increased. Steam sterilization (121 ° C., 15 minutes) was performed. An aqueous solution containing 1 mL of 1 M potassium dihydrogen phosphate buffer (buffer adjusted to pH 7.0 by mixing 1 M aqueous solution of dipotassium dihydrogen phosphate and 1 M aqueous solution of dipotassium hydrogen phosphate) and 0.1 M of L-arginine in 100 mL of this aqueous solution. 1 mL, 2 mL of an aqueous solution containing 50% by weight of 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, and the pore size was 0.22 μm. AB medium was obtained by performing filtration sterilization with a router.

HMF-containing liquid 4-hydroxy-5-methyl-3 (2H) -furanone (HMF) 2.35 mg, ethanol 1 mL, and sterile distilled water 9 mL were mixed in a glass container, and the HMF-containing liquid (2,000 μM HMF) was mixed. (Contains) was prepared. HMF is known to have AI-2 activity and HMF was used as an AI-2 substitute in this study.

-BMF-containing liquid 4-bromo-5- (4-methoxyphenyl) -2 (5H) -furanone (BMF) 2.15 mg, ethanol 4 mL, and sterile distilled water 36 mL are mixed, and a BMF-containing liquid (containing 200 μM BMF) is mixed. ) Was prepared. BMF is known to have an activity of inhibiting AI-2, and in this study, BMF was used as a positive control.

-Test solution Add sodium copper chlorophyllin to an aqueous solution containing 10% by volume of ethanol so as to be 200 μM, 400 μM, 1000 μM, 1400 μM, 2000 μM, 3000 μM, 4000 μM, 10000 μM, 16000 μM, 20000 μM, 30000 μM, 40,000 μM, and 50000 μM. Test solutions of various concentrations were prepared.

(2) Measurement conditions Preculture the reporter bacteria in Km-LB medium, _{measure the 600} nm absorbance (OD 600) of the preculture solution, and add an appropriate amount of Km-LB medium so that OD ₆₀₀ becomes 0.50. The reporter's bacterial solution was obtained. This reporter bacterial solution was diluted 2,500 times with AB medium to obtain a reporter bacterial solution.

In a sterile glass test tube, 0.9 mL of the reporter bacterial diluted solution and 0.05 mL of the test solution were mixed and incubated for 10 minutes under the conditions of 30 ° C. and stirring (180 rpm) under shading. Then, 0.05 mL of the HMF-containing solution was added, and the mixture was incubated under shading at 30 ° C. and stirring (180 rpm) for 4 hours. Since the test solution was diluted 20 times at this point, the final concentrations of sodium copper chlorophyllin 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 It is 2500 μM. 0.1 mL of the liquid after incubation was transferred to a luminescence measurement plate, and the luminescence intensity (sample luminescence intensity) of 578 nm was measured with a microplate reader (device: SpectraMax iD3, measurement mode: LUM endpoint, MOLECULAR DEVICE). In addition, the viable number of reporter bacteria in the solution after incubation was measured by the colony counting method.

As a blank, an aqueous solution containing 10% by volume ethanol was used instead of the test solution, and an aqueous solution containing 10% by volume ethanol was used instead of the HMF-containing solution, and the emission intensity (of the blank) was the same as described above. Emission intensity) and viable cell count were measured.

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

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

The AI-2 inhibition rate was calculated from the measured emission intensity values according to the following formula.

The obtained results are shown in FIG. As a result, it was clarified that sodium copper chlorophyllin does not show antibacterial activity in the concentration range of 10 to 2500 μM and has an activity of effectively inhibiting AI-2. In particular, it was confirmed that copper chlorophyllin sodium showed remarkably excellent AI-2 inhibitory activity in the concentration range of 100 to 2500 μM.

Test Example 2: in an aqueous solution containing a verification 10% by volume of ethanol AI-2 inhibitory activity of vitamin B _2, 400 [mu] M vitamin B ₂ (sodium riboflavin phosphate), 1000μM, 2000μM, 3000μM, 4000μM, 10000μM, and a 16000μM The test solutions having various concentrations were prepared. Using the obtained test solution, the AI-2 inhibition rate and the viable cell count were measured under the same conditions as in Test Example 1. _{That is, in this test, the final concentrations of vitamin B 2} at the time of measuring the AI-2 inhibition rate and the viable cell count were 20 μM, 50 μM, 100 μM, 150 μM, 200 μM, 500 μM, and 800 μM.

The obtained results are shown in FIG. As a result, _{it was clarified that vitamin B 2} did not show antibacterial activity in the concentration range of 20 to 800 μM and had an activity of effectively inhibiting AI-2. In particular, _{it was confirmed that vitamin B 2} showed remarkably excellent AI-2 inhibitory activity in the concentration range of 100 to 800 μM.

Test Example 3: Verification of AI-2 Inhibitory Activity of Tin Fluoride To an aqueous solution containing 10% by volume of ethanol, tin (II) fluoride was added at 200 μM, 1000 μM, 2000 μM, and 4000 μM to test various concentrations. A solution was prepared. Separately, in a bentonite dispersion prepared by dispersing bentonite (Benton DE, manufactured by Kirest Co., Ltd.) in sterile distilled water, tin (II) fluoride is 10000 μM, 20000 μM, 40,000 μM, 70000 μM, 80000 μM, and 90000 μM. To prepare test solutions of various concentrations. Using the obtained test solution, the AI-2 inhibition rate and the viable cell count were measured under the same conditions as in Test Example 1. That is, in this test, the final concentrations of tin 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. In this test, an aqueous solution containing 10% by volume of ethanol or a bentonite dispersion was used for the measurement of blank and negative control.

The obtained results are shown in FIG. As a result, it was clarified that tin fluoride did not show antibacterial activity in the concentration range of 10 to 4500 μM and had an activity of effectively inhibiting AI-2. In particular, it was confirmed that tin fluoride showed remarkably excellent AI-2 inhibitory activity in the concentration range of 100 to 4500 μM.

Test Example 4: Verification of AI-2 Inhibitory Activity of Zinc Chloride Zinc chloride is added to an aqueous solution containing 10% by volume of ethanol so as to be 400 μM, 1000 μM, 4000 μM, 10000 μM, 16000 μM, 20000 μM, 30000 μM and 40,000 μM, and various types are added. A test solution having a concentration was prepared. Using the obtained test solution, the AI-2 inhibition rate and the 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 at the time of measuring the AI-2 inhibition rate and the viable cell count are 20 μM, 50 μM, 200 μM, 500 μM, 800 μM, 1000 μM, 1500 μM and 2000 μM.

The obtained results are shown in FIG. As a result, it was clarified that zinc chloride does not show antibacterial activity in the concentration range of 20 to 2000 μM and has an activity of effectively inhibiting AI-2. In particular, it was confirmed that zinc chloride exhibits remarkably excellent AI-2 inhibitory activity in the concentration range of 50 to 2000 μM.

Test Example 5: Verification of AI-2 inhibitory activity of aluminum lactate In an aqueous solution containing 10% by volume of ethanol, aluminum lactate was added to 2000 μM, 3000 μM, 4000 μM, 6000 μM, 10000 μM, 20000 μM, 30000 μM, 40,000 μM, 50000 μM, 60000 μM, and 70000 μM. The test solutions having various concentrations were prepared. Using the obtained test solution, the AI-2 inhibition rate and the 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 the viable cell count were 100 μM, 150 μM, 200 μM, 300 μM, 500 μM, 1000 μM, 1500 μM, 2000 μM, 2500 μM, 3000 μM, and 3500 μM. ..

The obtained results are shown in FIG. As a result, it was clarified that aluminum lactate does not show antibacterial activity in the concentration range of 100 to 3500 μM and has an activity of effectively inhibiting AI-2. In particular, it was confirmed that aluminum lactate exhibits remarkably excellent AI-2 inhibitory activity in the concentration range of 2000 to 3500 μM.

Claims (7)

An autoinducer-2 inhibitor containing at least one selected from the group consisting of copper chlorophyllin alkali metal salt, vitamin B ₂ , tin fluoride, zinc chloride, and aluminum lactate as an active ingredient. The active ingredient is copper chlorophyllin alkali metal salt,
The auto-inducer-2 inhibitor according to claim 1, which is used so that the concentration of the copper chlorophyllin alkali metal salt is 10 to 2500 μM at the site where inhibition of auto-inducer-2 is required. The active ingredient is vitamin B ₂
The auto-inducer-2 inhibitor according to claim 1, which is used so that the concentration _{of vitamin B 2} is 20 to 800 μM at a site where inhibition of auto-inducer-2 is required. The active ingredient is tin fluoride
The auto-inducer-2 inhibitor according to claim 1, which is used so that the concentration of tin fluoride is 10 to 4500 μM at a site where inhibition of auto-inducer-2 is required. The active ingredient is zinc chloride
The auto-inducer-2 inhibitor according to claim 1, which is used so that the concentration of zinc chloride is 20 to 2000 μM at a site where inhibition of auto-inducer-2 is required. The active ingredient is aluminum lactate.
The auto-inducer-2 inhibitor according to claim 1, which is used so that the concentration of aluminum lactate is 100 to 3500 μM at a site where inhibition of auto-inducer-2 is required. The auto-inducer-2 inhibitor according to any one of claims 1 to 6, which is an oral care product.

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