Promoter for proliferation of nitrate-reducing bacteria in oral cavity

Abstract

To provide a promoter for proliferation of nitrate-reducing bacteria in the oral cavity, which enables selective proliferation of nitrate-reducing bacteria in the oral cavity.SOLUTION: A promoter for proliferation of nitrate-reducing bacteria in the oral cavity comprises one or more selected from the group consisting of glycyrrhetinic acid, glycyrrhizic acid, and salts thereof. Preferably, the nitrate-reducing bacteria to be proliferated include one or more species selected from the group consisting of bacteria of the genus Neisseria and bacteria of the genus Haemophilus. The promoter further comprises a nitrate ion-supplying compound. The promoter serves as an agent for preventing or improving dental caries or an agent for periodontal disease.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to an agent for promoting the growth of nitrate-reducing bacteria in the oral cavity.

Background Art

[0002] There are more than 700 species of bacteria in the oral cavity. In recent years, it has been reported that the balance of the oral microbiota is disrupted in patients with dental caries and periodontal disease compared to healthy individuals, and it has come to be considered that controlling the oral microbiota is important for preventing dental caries and periodontal disease. Furthermore, with the progress of analysis techniques using bacteria-derived DNA, understanding has advanced not only for bacteria that cause diseases but also for bacteria related to health. For example, bacteria of the genus Neisseria and Haemophilus are representative bacteria that are abundant in healthy individuals and have the ability to reduce nitrate ions (NO3 - ) to nitrite ions (NO2 - ) or nitric oxide (NO). As mechanisms by which bacteria having nitrate-reducing ability, i.e., nitrate-reducing bacteria, bring benefits to the host human, there are known an action of lowering blood pressure by nitric oxide, an antibacterial action against periodontal pathogenic bacteria, and an action of preventing tooth demineralization by suppressing a decrease in saliva pH. Therefore, selectively growing nitrate-reducing bacteria in the oral cavity is useful for preventing dental caries and periodontal disease.

[0003] Since nitrate-reducing bacteria use nitrate as a main energy source, a high concentration of nitrate must be supplied for nitrate-reducing bacteria to grow in the oral cavity. On the other hand, it has also been pointed out that excessive intake of nitrate is associated with a risk of generating carcinogenic N-nitroso compounds. Therefore, even under the situation where the oral nitrate concentration is low, a means for growing nitrate-reducing bacteria is desired. Thus, an investigation was started to search for an agent for promoting the growth of nitrate-reducing bacteria.

[0004] Patent Document 1 describes that an oral care composition containing a water-soluble nitrate, a basic amino acid, a water-soluble alkali metal polyphosphate, and water can promote the nitrate reduction of the oral microbiome.

Prior Art Documents

Patent Documents

[0005] [Patent Document 1] International Publication No. 2022 / 251626 [Summary of the Invention] [Problems to be Solved by the Invention]

[0006] However, there is no known component having an effect of selectively growing specific oral bacteria.

[0007] An object of the present invention is to provide an oral nitrate-reducing bacterium growth agent capable of selectively growing nitrate-reducing bacteria in the oral cavity. [Means for Solving the Problems]

[0008] The present invention provides the following [1] to [5]. [1] An oral nitrate-reducing bacterium growth agent containing one or more selected from the group consisting of glycyrrhetinic acid, glycyrrhizic acid, and salts thereof. [2] The agent according to [1], wherein the nitrate-reducing bacteria include one or more selected from the group consisting of bacteria of the genus Neisseria and bacteria of the genus Haemophilus. [3] The agent according to [2], satisfying at least one of the following: the bacteria of the genus Neisseria include one or more selected from the group consisting of Neisseria mucosa and Neisseria flavescens; and the bacteria of the genus Haemophilus include Haemophilus parainfluenzae. [4] The agent according to any one of [1] to [3], further containing a nitrate ion supply compound. [5] The agent according to any one of [1] to [4], which is a caries prevention / improvement agent or a periodontal disease prevention / improvement agent. [Effects of the Invention]

[0009] According to the present invention, since nitrate-reducing bacteria in the oral cavity can be selectively grown, the nitrate-reducing ability in the oral cavity is enhanced, and caries and periodontal diseases can be prevented or improved by nitrite and nitric oxide.

Mode for Carrying Out the Invention

[0010] [1. Oral nitrate-reducing bacteria growth promoter] [Glycyrrhetinic acids] The oral nitrate-reducing bacteria growth promoter contains at least one selected from the group consisting of glycyrrhetinic acid, glycyrrhizic acid and salts thereof (hereinafter sometimes abbreviated as glycyrrhetinic acids). A combination of two or more of these may be contained.

[0011] Examples of glycyrrhetinic acid include α-glycyrrhetinic acid and β-glycyrrhetinic acid, and β-glycyrrhetinic acid is preferable.

[0012] Glycyrrhizic acid exists as a glycyrrhizic acid salt. Examples of the glycyrrhizic acid salt include dipotassium glycyrrhizate, diammonium glycyrrhizate, disodium glycyrrhizate, trisodium glycyrrhizate, and monoammonium glycyrrhizate, and dipotassium glycyrrhizate is preferable.

[0013] Glycyrrhetinic acid, glycyrrhizic acid and salts thereof may be any of natural raw materials (natural) themselves, extracts or purified products obtained from the above raw materials, and chemically synthesized products. Examples of natural raw materials include the plant body of licorice (e.g., Ural licorice, Spanish licorice), a part thereof (e.g., roots, rhizomes), or processed products thereof (e.g., leachate, extract (water and / or ethanol extract), concentrated extract, powder).

[0014] [Nitrate ion supply compound] The oral nitrate-reducing bacteria growth promoter preferably further contains a nitrate ion supply compound. Thereby, nitrate ions utilized by nitrate-reducing bacteria can be supplied.

[0015] Examples of the nitrate ion supply compound include nitrate ions (NO3 -Any compound that can supply is acceptable. Examples include nitrate esters and nitrates, with nitrates being preferred. Examples of nitrate salts include counterions of nitrate ions such as alkali metal salts (e.g., sodium salt, potassium salt), alkaline earth metal salts (e.g., calcium salt), and ammonium salts. Nitrates are preferably potassium nitrate, sodium nitrate, calcium nitrate, and ammonium nitrate, more preferably potassium nitrate, sodium nitrate, and ammonium nitrate, and even more preferably potassium nitrate.

[0016] The nitrate ion supply compound can be either derived from natural raw materials (natural) containing the nitrate ion supply compound or a chemically synthesized product containing the nitrate ion supply compound, but a chemically synthesized product is preferred. Examples of natural raw materials include vegetables or fruits. More specifically, they include beets, spinach, green and yellow vegetables (e.g., Chinese cabbage, spinach, broccoli, komatsuna), light-colored vegetables (e.g., cabbage, lettuce, celery), combinations of two or more of these, and processed products of these (e.g., powders).

[0017] The nitrate ion supply compound can be used alone or in combination of two or more.

[0018] [The effective amount of glycyrrhetinic acids and the nitrate ion supply compound] The effective amount (mass fraction) of glycyrrhetinic acids with respect to the oral nitrate-reducing bacteria growth agent is preferably 50 ppm or more, more preferably 100 ppm or more, and even more preferably 150 ppm or more. The upper limit is not particularly limited, but is usually 3000 ppm or less, preferably 2200 ppm or less, and more preferably 2100 ppm or less. Therefore, the effective amount of glycyrrhetinic acids is preferably 50 ppm or more, more preferably 50 - 3000 ppm, even more preferably 100 - 2200 ppm, and even more preferably 150 - 2100 ppm. When the effective amount of glycyrrhetinic acids with respect to the oral nitrate-reducing bacteria growth agent satisfies the above numerical range, the agent of the present invention can more preferably exhibit a selective nitrate-reducing bacteria growth effect. The "effective amount" refers to the effective amount of component (A) in the saliva secreted into the oral cavity assuming a saliva secretion rate of 1 mL / min. The same applies to the effective amount of component (B) below.

[0019] - Nitrate ion supply compound - The effective amount (mass fraction) of the nitrate ion supply compound with respect to the oral nitrate-reducing bacteria growth promoter is preferably 5 to 10,000 ppm, more preferably 10 to 5,000 ppm. Nitrate ions are usually present in saliva, but by adding the nitrate ion supply compound to the agent of the present invention so as to satisfy the above numerical range, the effect of promoting the growth of oral nitrate-reducing bacteria is further enhanced.

[0020] When the oral nitrate-reducing bacteria growth promoter contains both glycyrrhetinic acids and a nitrate ion supply compound, the effective amount ratio of glycyrrhetinic acids to the nitrate ion supply compound is preferably 0.005 or more, more preferably 0.005 to 300, and still more preferably 0.02 to 220. By satisfying the above numerical range for the effective amount ratio of glycyrrhetinic acids to the nitrate ion supply compound, the effects of the present invention can be more preferably exhibited.

[0021] [2. Oral nitrate-reducing bacteria growth effect] The oral nitrate-reducing bacteria growth promoter can exhibit an oral nitrate-reducing bacteria growth effect. In this specification, the oral nitrate-reducing bacteria growth effect means increasing or maintaining at an appropriate level the ratio of the number of oral nitrate-reducing bacteria to the total number of bacteria in the oral cavity and / or the number of oral nitrate-reducing bacteria.

[0022] - Oral nitrate-reducing bacteria - The oral nitrate-reducing bacteria may be any microorganisms that are present in the oral cavity and have nitrate-reducing ability. For example, bacteria of the genus Neisseria, bacteria of the genus Haemophilus, bacteria of the genus Veillonella, bacteria of the genus Granulicatella, bacteria of the genus Rothia, bacteria of the genus Rothia, bacteria of the genus Actinomyces, bacteria of the genus Leptotrichia, and bacteria of the genus Scardovia can be mentioned. These bacteria are more numerous in healthy individuals compared to periodontal disease patients and are bacteria with nitrate-reducing ability.

[0023] Examples of Neisseria bacteria include, for example, Neisseria mucosa, Neisseria flavescens, Neisseria subflava, Neisseria denitrificans, and Neisseria sicca. Preferably, they are Neisseria mucosa and Neisseria flavescens. Examples of Haemophilus bacteria include, for example, Haemophilus parainfluenzae, Haemophilus haemoglobinophilus, and Haemophilus haemolyticus. Preferably, it is Haemophilus parainfluenzae. Examples of Veillonella bacteria include, for example, Veillonella parvula. Examples of Granullicatella bacteria include, for example, Granullicatella adiacens. Examples of Lautropia bacteria include, for example, Lautropia mirabilis. Examples of Rothia bacteria include, for example, Rothia mucilaginosa, Rothia dentocariosa, and Rothia aeria. Examples of Actinomyces bacteria include, for example, Actinomyces viscosus and Actinomyces naeslundii. Examples of the genus Leptotrichia bacteria include, for example, Leptotrichia buccalis. Examples of the genus Schaalia bacteria include, for example, Schaalia odontolytica.

[0024] Among these, preferably, the bacteria are of the genus Neisseria and the genus Haemophilus, more preferably Neisseria mucosa, Neisseria flavescens, and Haemophilus parainfluenzae. Thereby, in the oral cavity, the ability to reduce nitrate to nitrite or nitric oxide is further enhanced, which is more suitable for the prevention or improvement of dental caries and periodontal disease.

[0025] In the agent of the present invention, as the nitrate-reducing bacteria, any one of the above-mentioned bacteria may be used alone, or a combination of two or more may be used.

[0026] The oral cavity nitrate-reducing bacteria growth agent can exert a growth effect on all oral cavity nitrate-reducing bacteria, and is also useful as an oral cavity nitrate-reducing bacteria growth agent for Neisseria mucosa, Neisseria flavescens, or Haemophilus parainfluenzae. That is, the oral cavity nitrate-reducing bacteria growth agent can also exert a good effect on each individual bacterial species among the nitrate-reducing bacteria.

[0027] It is known that nitrite ions and nitric oxide reduced by bacteria of the genus Neisseria or the genus Haemophilus have an antibacterial action against periodontal pathogenic bacteria and dental caries pathogenic bacteria, and an action of preventing tooth demineralization by suppressing the decrease in saliva pH. In addition, nitrite is considered to produce nitric oxide (NO) in tissues such as the stomach, and exert a blood pressure lowering action due to the effect of improving vascular function and an action of improving motor function due to the effect of improving skeletal muscle contractility. Therefore, the above-mentioned glycyrrhetinic acids can exert effects associated with the nitrate-reducing bacteria growth effect. For example, prevention or improvement of periodontal disease, prevention or improvement of dental caries, increase in nitric oxide, prevention or improvement of hypertension, and improvement or enhancement of motor function can be mentioned.

[0028] In addition, the inhibitory effect of Neisseria mucosa on the infection of periodontal pathogenic bacteria to gingival cells is known. When an oral nitrate-reducing bacteria growth promoter is used as the oral composition described below, the inhibitory effect of Neisseria mucosa can suppress the amount of bactericide used, and a more highly safe preparation can be obtained.

[0029] - Administration method and target - Examples of the administration method of the agent include oral administration (for example, intraoral administration, sublingual administration), parenteral administration (for example, transdermal administration, intravenous administration, intramuscular administration, subcutaneous administration, nasal administration, pulmonary administration). Among these, an administration form with less invasiveness is preferable, transdermal administration (for external use), oral administration (for internal use) is more preferable, oral administration is even more preferable, and mucosal administration (for external use on mucosa) is even more preferable.

[0030] The administration target may be an animal including humans, and is usually a human. The administration target may be a healthy person, but may also be a person infected with a bacterial infection in the oral cavity (for example, periodontal disease, dental caries, halitosis) or a person suspected of being infected. Examples of animals other than humans include mammals such as mice, rats, hamsters, dogs, cats, sheep, goats, cows, pigs, and monkeys.

[0031] [3. Use as pharmaceuticals, quasi-drugs, and cosmetics] The oral nitrate-reducing bacteria growth promoter can be used as a pharmaceutical, a quasi-drug, or a cosmetic. Examples of the dosage form include a liquid agent, a spray agent, a solid agent, a semi-solid agent, a liquid agent, a powder agent, and a granule agent, which can be determined according to various uses.

[0032] [4. Oral composition] The agent containing the above glycyrrhetinic acids can be used as an oral composition. Thereby, the oral nitrate-reducing bacteria growth effect can be efficiently exhibited. In this specification, the oral composition means a composition applied to the oral cavity, regardless of whether it is a pharmaceutical, a quasi-drug, or a cosmetic. For example, dentifrices (for example, paste dentifrice, gel dentifrice, moist dentifrice, liquid dentifrice), mouthwashes, tongue cleaners, oral sprays, oral tablets, oral films, gums, oral cooling agents, gargle tablets, oral pastes, gel agents, and ointment agents can be mentioned.

[0033] Among these, preferably they are pharmaceuticals or quasi-drugs, more preferably dentifrices or mouthwashes.

[0034] [5. Dosage form] Examples of dosage forms include, for example, liquid (solution), syrup-like (syrup), cream-like, paste-like, tablet (tablet, tablet), capsule-like (capsule), powder-like (granule, fine granule), film-like, soft capsule-like (soft capsule based on gelatin, etc.), hard capsule-like (hard capsule), and they may be appropriately selected according to the administration form.

[0035] [Optional components] The oral composition may contain other optional components other than glycyrrhetinic acids. Examples of other components include, for example, bactericides, preservatives, medicinal ingredients, surfactants, abrasives, wetting agents, binders, buffers, solubilizers, tonicity agents, stabilizers, chelating agents, moisturizing agents, flavoring agents (sweeteners, fragrances, acidulants), oily components, coloring agents, pH adjusters, solvents, excipients, disintegrants, binders, lubricants, color developers, antioxidants, fortifiers, swelling agents, thickeners, cooling agents, astringents, ultraviolet absorbers, aqueous solvents, etc., components other than the above active ingredients. The type and content of the optional components may be selected according to the uses of pharmaceuticals, quasi-drugs, cosmetics, and / or dosage forms, administration methods, etc., and may be one type or a combination of two or more types.

[0036] - Bactericide - Examples of bactericides include, for example, cetylpyridinium chloride, benzalkonium chloride, benzethonium chloride, alkyl diminoethyl glycine hydrochloride, chlorhexidine or its salts, triclosan, isopropylmethylphenol, hinokitiol, chlorhexidine gluconate, decalinium chloride, iodine, potassium iodide, sulfamethoxazole, sodium sulfamethoxazole, sulfisoxazole, sodium sulfisomidine.

[0037] - Preservative - Examples of the preservative include paraoxybenzoic acid esters such as methyl paraben, ethyl paraben, propyl paraben, and butyl paraben, benzoic acid or its salts, dehydroacetic acid or its salts, propionic acid or its salts, sorbic acid or its salts (such as potassium sorbate), boric acid, borax, alcohol derivatives such as phenylethyl alcohol, benzyl alcohol, phenol, and acrinol, and alkyl polyaminoethyl glycine.

[0038] In the present invention, it is more preferable not to contain the above-mentioned bactericide and / or preservative (each content is 0%) in terms of inhibiting the growth of nitrate-reducing bacteria. However, the bactericide and / or preservative may be contained as long as the effects of the present invention are not impaired. When contained, usually, the total amount of the bactericide and / or preservative is 0.1% by mass or less, preferably 0.05% by mass or less, more preferably 0.03% by mass or less, still more preferably 0.01% by mass or less, and even more preferably 0.005% by mass or less of the whole agent composition. Thereby, non-selective sterilization against pathogenic bacteria and resident bacteria can be suppressed, and the risk of dysbacteriosis due to the disruption of the bacterial balance in the oral cavity can be reduced.

[0039] -Medicinal ingredient- Examples of the medicinal ingredients include enzymes such as dextranase, mutanase, amylase, protease, and lytic enzyme; fluorides such as sodium fluoride, sodium monofluorophosphate, and stannous fluoride; anti-inflammatory agents such as tranexamic acid, epsilon-aminocaproic acid, allantoin, allantoin chlorhydroxyaluminum, azulene, and dihydrocholesterol; metal salts such as zinc salts, copper salts, and tin salts; calculus preventives such as condensed phosphates and ethanehydroxydiphosphonate; blood flow accelerators such as vitamin E (e.g., tocopherol acetate); hypersensitivity inhibitors such as strontium chloride; coating agents such as hydroxyethylcellulose dimethyldiallylammonium chloride; astringents such as vitamin C (e.g., ascorbic acid or its salts), lysozyme chloride, and sodium chloride; water-soluble copper compounds such as copper chlorophyll; calculus preventives; amino acids such as alanine, glycine, and proline; plant extracts such as thyme, saffron, cloves, and witch hazel; caropeptide; polyvinylpyrrolidone, etc. Other examples include decongestants, anti-inflammatory agents, astringents, antihistamines, vitamins, amino acids, bactericides, local anesthetics, components having a nitrate-reducing bacterium growth promoting effect other than the active ingredients in the present invention, and combinations of two or more selected from these. Examples of the decongestants include naphazoline hydrochloride, tetrahydrozoline hydrochloride, phenylephrine hydrochloride, epinephrine, epinephrine hydrochloride, ephedrine hydrochloride, and dl-methylephedrine hydrochloride. Examples of the anti-inflammatory and astringent agents include neostigmine methylsulfate, allantoin, berberine chloride, berberine sulfate, zinc sulfate, zinc lactate, lysozyme chloride, bromelain, methyl salicylate, sodium azulenesulfonate, chamomile, and sodium cromoglycate. Examples of the antihistamines include iproheptine hydrochloride, diphenhydramine hydrochloride, diphenhydramine, isothipendyl hydrochloride, and chlorpheniramine maleate. Examples of the vitamins include flavin adenine dinucleotide sodium, pyridoxine hydrochloride, cyanocobalamin, vitamin A compounds (e.g., retinol acetate, retinol palmitate), and vitamin E compounds (tocopherol acetate (e.g., d-α-tocopherol acetate)).Examples of the amino acids include potassium L-aspartate, magnesium L-aspartate, aminoethylsulfonic acid, and sodium chondroitin sulfate. Examples of the local anesthetics include lidocaine, lidocaine hydrochloride, dibucaine hydrochloride, and chlorobutanol. Each of the medicinal components may be used alone or in combination of two or more. The content of the medicinal component can be appropriately set to an effective amount according to a conventional method.

[0040] -Surfactant- Examples of the surfactant include nonionic surfactants, anionic surfactants, and amphoteric surfactants.

[0041] Examples of the nonionic surfactants include polyoxyethylene alkyl ether, polyoxyethylene hydrogenated castor oil, sorbitan fatty acid ester, polyoxyethylene sorbitan fatty acid ester (e.g., polyoxyethylene sorbitan monostearate), alkylolamide, polyoxyethylene fatty acid ester, polyglycerin fatty acid ester, sucrose fatty acid ester (e.g., maltose fatty acid ester), sugar alcohol fatty acid ester (e.g., maltitol fatty acid ester, lactitol fatty acid ester), fatty acid diethanolamide (e.g., lauric acid mono- or diethanolamide), polyoxyethylene polyoxypropylene copolymer, polyoxyethylene polyoxypropylene fatty acid ester, glycerin fatty acid ester, polyoxyethylene glycerin fatty acid ester, polyoxyethylene glycol fatty acid ester, polyoxyethylene phytosterol and phytostanol, polyoxyethylene lanolin and lanolin alcohol, polyoxyethylene alkylamine and fatty acid amide, polyoxyethylene alkyl phenyl formaldehyde condensate, polyoxyethylene polyoxypropylene alkyl ether, and polyoxyethylene alkyl phenyl ether. Among these, polyoxyethylene alkyl ether and polyoxyethylene hydrogenated castor oil are preferable, and polyoxyethylene hydrogenated castor oil is more preferable.

[0042] Examples of anionic surfactants include, for example, alkyl sulfates, acyl amino acid salts, acyl taurine salts, α-olefin sulfonates, hydrogenated coconut fatty acid monoglyceride monosulfates, and lauryl sulfacetates. The alkyl group and acyl group may be either linear or branched, and may be either saturated or unsaturated, and the number of carbon atoms thereof is usually 10 to 20, preferably 12 to 18, and more preferably 12 to 14. The salt can be selected from pharmacologically acceptable salts. Examples of pharmacologically acceptable salts include, for example, base addition salts and amino acid salts. Specific examples thereof include inorganic base salts such as sodium salt, potassium salt, calcium salt, magnesium salt, and ammonium salt; organic base salts such as triethylammonium salt, triethanolammonium salt, pyridinium salt, and diisopropylammonium salt; and basic amino acid salts such as arginine salt. Among them, inorganic base salts are preferred, alkali metal salts (for example, sodium salt, potassium salt) or ammonium salts are more preferred, and sodium salt is even more preferred. Examples of alkyl sulfates include, for example, myristyl sulfate. Examples of acyl amino acid salts include acyl glutamic acid salts such as lauroyl glutamate, myristoyl glutamate, and palmitoyl glutamate; acyl glycine salts such as N-lauroyl-N-methylglycine salt and cocoyl glycine salt; acyl alanine salts such as N-lauroyl-β-alanine salt, N-myristyl-β-alanine salt, N-cocoyl-β-alanine salt, N-lauroyl-N-methyl-β-alanine salt, N-myristoyl-N-methyl-β-alanine salt, and N-methyl-N-acylalanine salt; and acyl aspartic acid salts such as lauroyl aspartate. Examples of acyl taurine salts include, for example, N-methyl-N-acyl taurine salt and N-cocoyl methyl taurine salt. Examples of anionic surfactants also include sodium hydrogenated coconut fatty acid monoglyceride monosulfate and sodium lauryl sulfacetate.

[0043] Examples of amphoteric surfactants include betaine-type amphoteric surfactants such as alkyl dimethylaminoacetic acid betaines (e.g., lauryl dimethylaminoacetic acid betaine), fatty acid amide propyl dimethylaminoacetic acid betaine; imidazoline-type amphoteric surfactants such as N-fatty acyl-N-carboxymethyl-N-hydroxyethyl ethylenediamine salts, coconut oil fatty acid imidazolinium betaine; and alkyl betaines such as lauryl dimethylaminoacetic acid betaine.

[0044] When the composition contains a surfactant, the content of each of the anionic, nonionic, and amphoteric surfactants is usually 0.01 to 10% by mass, preferably 0.1 to 5% by mass, more preferably 0.2 to 3% by mass of the whole composition.

[0045] -Abrasive agent- As the abrasive agent, for example, either an inorganic abrasive agent or an organic abrasive agent may be used. Examples of inorganic abrasive agents include abrasive silica such as precipitated silica, aluminosilicate, zirconosilicate, crystalline zirconium silicate, titanium-bonded silica; calcium phosphate-based compounds such as dibasic calcium phosphate dihydrate or anhydrate, monobasic calcium phosphate, tribasic calcium phosphate, calcium pyrophosphate; calcium carbonate-based abrasives such as calcium carbonate; calcium-based abrasives other than carbonate / phosphate such as calcium hydroxide, calcium sulfate; aluminum-based materials such as aluminum oxide, aluminum hydroxide, alumina; silicate-based materials such as anhydrous silicic acid, zeolite, zirconium silicate; magnesium-based materials such as magnesium carbonate, tribasic magnesium phosphate; apatite-based materials such as hydroxyapatite, fluoroapatite, calcium-deficient apatite; titanium-based materials such as titanium dioxide, mica titanium, titanium oxide; and minerals such as bentonite. Examples of organic abrasive agents include polymethyl methacrylate and synthetic resin-based abrasive agents. Among these, abrasive silica and calcium phosphate-based compounds are preferred, and anhydrous silicic acid is more preferred. The amount of the abrasive agent is preferably 50% by mass or less, more preferably 8 to 50% by mass based on the whole composition.

[0046] -Wetting agent- Examples of the wetting agent include sugar alcohols and polyhydric alcohols other than sugar alcohols. Examples of the sugar alcohol include sorbitol (sorbit), erythritol, maltitol, lactitol, xylitol, etc. Examples of the polyhydric alcohol other than sugar alcohols include glycerin; glycols such as ethylene glycol, propylene glycol, dipropylene glycol, butylene glycol, polyethylene glycol (PEG); and reduced starch saccharides. As the polyethylene glycol, for example, polyethylene glycol having an average molecular weight of 150 to 6000 is preferable, and polyethylene glycol having an average molecular weight of 190 to 630 (PEG200, PEG300, PEG400, PEG600) is more preferable. The average molecular weight is the average molecular weight described in the 2006 Specifications for Quasi-drug Raw Materials. The content of the wetting agent is usually 40% by mass or less, preferably 1 to 30% by mass, based on the whole composition.

[0047] -Binder- Examples of the binder include any conventionally known suitable organic binder, such as polysaccharides, cellulose-based binders (e.g., carboxymethyl cellulose (CMC), hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, methyl cellulose, cationized cellulose, etc.), other polysaccharide thickeners (e.g., xanthan gum, guar gum, gellan gum, tragacanth gum, karaya gum, gum arabic, locust bean gum, carrageenan, sodium alginate), synthetic water-soluble polymers (e.g., sodium polyacrylate, carboxyvinyl polymer, polyvinyl pyrrolidone, polyvinyl alcohol, propylene glycol alginate). Further, inorganic binders such as thickening silica and aluminum silicate can also be contained. The content of the organic binder is preferably 0 to 3% by mass, more preferably 0.1 to 2% by mass, based on the whole composition. The content of the inorganic binder is preferably 0 to 10% by mass, more preferably 1 to 8% by mass.

[0048] -Buffer- Examples of buffers include citric acid or its salts (e.g., sodium citrate), phosphoric acid or its salts (e.g., sodium hydrogen phosphate, sodium dihydrogen phosphate, dipotassium phosphate, potassium dihydrogen phosphate), tartaric acid or its salts (e.g., sodium tartrate), gluconic acid or its salts (e.g., sodium gluconate), acetic acid or its salts (e.g., sodium acetate), carbonic acid or its salts (e.g., sodium bicarbonate), trometamol, amino acids (e.g., potassium aspartate, aminoethylsulfonic acid, glutamic acid, sodium glutamate), and combinations of two or more of these.

[0049] - Solubilizing agent - Examples of solubilizing agents include propylene glycol, polyethylene glycol, and combinations of two or more of these.

[0050] - Isotonic agent - Examples of isotonic agents include sodium chloride, potassium chloride, glycerin, and combinations of two or more of these.

[0051] - Stabilizing agent - Examples of stabilizing agents include sodium edetate, cyclodextrin, sulfite, citric acid or its salts, dibutylhydroxytoluene, ascorbic acid, and combinations of two or more of these.

[0052] - Chelating agent - Examples of chelating agents include sodium edetate, sodium citrate, and combinations of these.

[0053] - Humectant - Examples of humectants include glycerin, concentrated glycerin, sugar alcohols (e.g., sorbitol, xylitol, maltitol, mannitol, reduced maltose, reduced palatinose, erythritol, lactitol, isomalt), and combinations of two or more of these.

[0054] - Flavoring agent - Examples of flavoring agents include, for example, sweeteners (e.g., artificial sweeteners such as sodium saccharin, aspartame, stevia, stevioside, paramethoxycinnamic aldehyde, neohesperidin dihydrochalcone, perillartine, aspartylphenylalanine methyl ester, thaumatin, acesulfame potassium, sucralose, maltitol, sorbitol, mannitol, reduced starch syrup, reduced palatinose, xylitol, erythritol, lactitol, etc.); fragrances (e.g., natural essential oils such as anise oil, cassia oil, wintergreen oil, mastic oil, neroli oil (orange flower oil), lemongrass oil, jasmine oil, rose oil, iris oil, clove oil, sage oil, cardamom oil, rosemary oil, laurel oil, chamomile oil, basil oil, marjoram oil, lemon oil, orange oil, lime oil, yuzu oil, nutmeg oil, lavender oil, paracresol oil, vanilla oil, cinnamon oil, pimento oil, bay leaf oil, perilla oil, wintergreen oil, peppermint oil, lychee oil, etc.); menthol, carvone, cinnamic aldehyde, anethole, methyl salicylate, eugenol, linalool, limonene, menthone, menthyl acetate, citral, decanal, camphor, borneol, pinene, spiranthol, n-decyl alcohol, citronellol, α-terpineol, citronellyl acetate, ethyl linalool, vanillin, etc., which are fragrance components contained in the above natural essential oils; ethyl acetate, ethyl butyrate, isoamyl acetate, hexanal, hexanal, methyl anthranilate, ethyl methylphenyl glycidate, benzaldehyde, vanilla, ethyl vanillin, furaneol, N-ethyl-p-menthane-3-carboxamide, menthyl lactate, ethylene glycol-l-menthyl carbonate, etc., which are fragrance components; and various compounded flavors such as mint-based, fruit-based, and herb-based flavors formed by combining several fragrance components and natural essential oils (e.g., peppermint micron X-8277-T, dry coat matcha #421), acidulants (e.g., citric acid, tartaric acid, malic acid), and green tea powder.

[0055] - Oil-based components - Examples of the oily component include fatty acid esters (e.g., glycerin fatty acid esters), hydrocarbons (e.g., paraffin, liquid paraffin, ceresin, squalane, petrolatum, microcrystalline wax), higher fatty acids (e.g., fatty acids having 8 to 22 carbon atoms such as lauric acid, myristic acid, oleic acid, isostearic acid), higher alcohols (e.g., alcohols having 8 to 22 carbon atoms such as lauryl alcohol, cetyl alcohol, cetostearyl alcohol, oleyl alcohol, isostearyl alcohol), vegetable oils and fats (e.g., vegetable oils such as olive oil, castor oil, coconut oil; fatty acid esters such as isopropyl myristate), beeswax, and combinations of two or more of these.

[0056] - Humectant - Examples of the humectant include sugar alcohols and polyhydric alcohols other than sugar alcohols. Examples of the sugar alcohol include sorbitol (sorbit), erythritol, maltitol, lactitol, xylitol, etc. Examples of the polyhydric alcohol other than sugar alcohols include glycerin; glycols such as ethylene glycol, propylene glycol, dipropylene glycol, butylene glycol, polyethylene glycol (PEG); and reduced starch saccharides. As the polyethylene glycol, for example, polyethylene glycol having an average molecular weight of 150 to 6000 is preferable, and polyethylene glycol having an average molecular weight of 190 to 630 (PEG200, PEG300, PEG400, PEG600) is preferable. The average molecular weight is the average molecular weight described in the Specifications for Quasi-Drug Raw Materials 2006. The amount of the humectant is usually 40% by mass or less, preferably 1 to 30% by mass, based on the whole composition.

[0057] - Colorant - Examples of the colorant include natural pigments such as safflower red pigment, gardenia yellow pigment, gardenia blue pigment, perilla pigment, red yeast rice pigment, red cabbage pigment, carrot pigment, hibiscus pigment, cocoa pigment, spirulina blue pigment, tamarind pigment, etc., legal pigments such as Red No. 2, Red No. 3, Red No. 104, Red No. 105, Red No. 106, Red No. 227, Yellow No. 4, Yellow No. 5, Green No. 3, Blue No. 1, etc., riboflavin, copper chlorophyllin sodium, and titanium dioxide. When a colorant is included, its content is preferably 0.00001 to 3% by mass based on the whole composition.

[0058] -pH adjuster- Examples of the pH adjuster include organic acids such as phthalic acid, citric acid, succinic acid, acetic acid, fumaric acid, malic acid, and lactic acid or their salts (sodium citrate), inorganic acids such as phosphoric acid (orthophosphoric acid) or their salts (for example, potassium salt, sodium salt, and ammonium salt), hydroxides such as sodium hydroxide and potassium hydroxide. Examples of the inorganic acid salt include disodium hydrogen phosphate and sodium dihydrogen phosphate. The content of the pH adjuster can usually be an amount such that the pH of the composition after addition is 5 to 9, preferably 6 to 8.5. In this specification, the pH value usually refers to the value 25 °C and 3 minutes after the start of measurement. The pH value can be measured, for example, using a pH meter (model number Hm-30S) manufactured by Toa Denpa Kogyo Co., Ltd.

[0059] -Solvent- Examples of the solvent include water (purified water) and ethanol, and water is preferred. The solvent may be used alone or in combination of two or more.

[0060] -Excipient- Examples of excipients include cellulose such as hydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose, crystalline cellulose, ethylcellulose, methyl ethylcellulose, low-substituted hydroxypropyl cellulose, and pharmacologically acceptable derivatives thereof; synthetic polymers such as polyvinylpyrrolidone and partially hydrolyzed polyvinyl alcohol; polysaccharides such as gelatin, gum arabic powder, pullulan, agar, alginic acid, sodium alginate, and chitosan gum; starches such as corn starch, potato starch, pregelatinized starch, and hydroxypropyl starch, and pharmacologically acceptable derivatives thereof; lactose, lactose granules, fructose, glucose, sucrose, granulated sugar, hydrated glucose, trehalose, palatinose, mannitol, sorbitol, erythritol, xylitol, maltotetraose, lactitol, isomalt, reduced palatinose, reduced maltose syrup, powdered reduced maltose syrup, and maltitol; inorganic excipients such as magnesium carbonate, calcium carbonate, light anhydrous silicic acid, silicon dioxide (also known as anhydrous silicic acid, fine particle silicon dioxide), titanium oxide, and aluminum hydroxide gel; and combinations of two or more of these.

[0061] - Disintegrant - Examples of disintegrants include crospovidone, calcium carboxymethylcellulose, croscarmellose sodium, low-substituted hydroxypropyl cellulose, carboxymethylcellulose, sodium carboxymethyl starch, croscarmellose sodium, cross-linked insoluble polyvinylpyrrolidone, hydroxypropyl starch, partially pregelatinized starch, corn starch, and combinations of two or more of these.

[0062] - Binder - Examples of binders include hydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose, ethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, gelatin, dextrin, starch, and pregelatinized starch, and combinations of two or more of these.

[0063] - Lubricant - Examples of the lubricant include calcium stearate, magnesium stearate, sucrose fatty acid ester, light anhydrous silicic acid, sodium stearyl fumarate, polyethylene glycol, talc, stearic acid, and combinations of two or more of these.

[0064] -Other optional components- Examples of the optional components other than those described above include polyisobutylene, polybutadiene, urethane, silicon, and natural rubber. The content of these other optional components can be appropriately set within a range that does not interfere with the effects of the present invention.

[0065] [6. Manufacturing method] The method for manufacturing an oral nitrate-reducing bacterium growth agent and an oral composition containing the same may be determined according to the dosage form, use, and application site. For example, when used as a dentifrice, a method may be adopted in which components soluble in a solvent are prepared, and then other insoluble components are mixed, and defoaming (e.g., under reduced pressure) is performed as necessary. As another example, an effective ingredient and other components used as necessary are dispersed and dissolved in an aqueous solvent (e.g., water such as purified water, sterilized water), and the composition is prepared and filled into an appropriate container (e.g., made of glass or resin). Examples of the container for an oral preparation include a laminated tube, and the material may be a resin such as polyethylene, polypropylene, polyethylene terephthalate, or nylon. In the case of a spray agent, a container equipped with a spraying means (e.g., a trigger-type, pump-type, or aerosol-type container) may be selected. The obtained dentifrice can be contained in a container to make a product. The shape and material of the container are not particularly limited, and a container commonly used for an oral composition can be used.

[0066] [7. Usage method] The method of using the oral nitrate-reducing bacteria growth promoter may be, for example, to administer the agent to the application site. The number of administrations per day is not particularly limited, and may be, for example, 1 to 6 times or more. In the case of an oral agent, for example, an appropriate amount of the agent is placed on a toothbrush and the tooth surface is brushed, and then rinsed with water after use (toothpaste), an appropriate amount of the agent is held in the mouth and gargled and then spit out (mouthwash), an appropriate amount of the agent is chewed and swallowed (oral tablet), or an appropriate amount of the agent is sprayed into the oral cavity (oral spray).

Examples

[0067] Hereinafter, the present invention will be described with reference to examples. The following examples do not limit the present invention.

[0068] (1) Preparation of bacterial solution Each strain shown in Table 1 was inoculated into Todd Hewitt Broth medium (Becton Dickinson) containing 5 μg / mL of hemin (FUJIFILM Wako Pure Chemical Corporation), 1 μg / mL of menadione (Sigma-Aldrich), 5% sterile defibrinated horse blood (Japan BioMaterials Center), and 1.5% Agar (Becton Dickinson), and aerobically cultured (37°C, 72 hours).

[0069] The grown colonies were suspended in 4 mL of Todd Hewitt Broth medium (Becton Dickinson) containing 5 μg / mL of hemin (FUJIFILM Wako Pure Chemical Corporation), 1 μg / mL of menadione (Sigma-Aldrich), and 0.5% Yeast Extract (Gibco), and aerobically cultured (37°C, 24 hours). For Haemophilus parainfluenzae, in addition to the above, it was aerobically cultured (37°C, 24 hours) using a medium supplemented with 1 mg / mL of β-nicotinamide adenine dinucleotide hydrate (Roche). Further, 2.5% of this culture solution was taken into 20 mL of the same medium and aerobically cultured (37°C, 24 hours) for use in the test.

[0070] (2) Confirmation of growth promotion effect The culture broth obtained in (1) was centrifuged at 13,000×g for 5 min, and the precipitate was resuspended in 2×Todd Hewitt Broth medium (Becton Dickinson) containing 10 μg / mL hemin (FUJIFILM Wako Pure Chemical Corporation) and 2 μg / mL menadione (Sigma-Aldrich) to a turbidity (OD550) of 1.0. For Haemophilus parainfluenzae, in addition to the above, the medium supplemented with 2 mg / mL β-nicotinamide adenine dinucleotide hydrate (Roche) was used for resuspension to a turbidity (OD550) of 1.0. Further, the adjusted bacterial suspension was diluted 50-fold using the same medium.

[0071] 100 μL of the adjusted bacterial suspension was added to a 96-well plate (Sumitomo Bakelite). Then, 100 μL of β-glycyrrhetinic acid or dipotassium glycyrrhizinate adjusted to the concentrations shown in Tables 2 and 3 was added, and aerobic culture was performed (37 °C, 24 hours) (Examples 1 to 8). For the control, 100 μL of water was added (Comparative Examples 1 and 2). For the blank, a medium not containing the bacterial suspension was used and adjusted in the same manner as the evaluation sample.

[0072] [Raw materials used] Dipotassium glycyrrhizinate (External standard dipotassium glycyrrhizinate standard product, manufactured by Maruzen Pharmaceutical Co., Ltd.) β-Glycyrrhetinic acid (External standard β-glycyrrhetinic acid (manufactured by Maruzen Pharmaceutical Co., Ltd.))

[0073] (3) Evaluation method The turbidity (OD550) of the 96-well plate cultured in (2) was measured using a plate reader (Tecan Japan). The samples evaluated for β-glycyrrhetinic acid were diluted 20-fold using the adjusted medium and then the turbidity (OD550) was measured. The growth promotion ratio was calculated according to the following formula, where the turbidity of the samples added with β-glycyrrhetinic acid and dipotassium glycyrrhizinate is a, the turbidity of the control is b, the blank of the sample is c, and the blank of the control is d. Growth promotion ratio = (a - c) / (b - d)

[0074]

Table 1

[0075]

Table 2

[0076]

Table 3

[0077] In Examples 1 to 8 containing dipotassium glycyrrhizinate or β-glycyrrhetinic acid, the turbidity of each nitrate-reducing bacterium in the culture plate was higher than that in Comparative Examples 1 and 2 which did not contain any of them, and the growth promotion ratio was high. Also, in any of the examples, the turbidity and growth promotion ratio of S. salivarius were comparable to or lower than those of the comparative examples, and the influence on the resident bacteria was relatively small. These results indicate that the agent of the present invention can act selectively on nitrate-reducing bacteria present in the oral cavity and exhibit a growth effect.

Claims

1. An oral nitrate-reducing bacteria growth agent containing one or more selected from the group consisting of glycyrrhetinic acid, glycyrrhizic acid, and salts thereof.

2. The agent according to claim 1, wherein the nitrate-reducing bacteria include one or more selected from the group consisting of bacteria of the genus Neisseria and bacteria of the genus Haemophilus.

3. The agent according to claim 2, wherein the bacteria of the genus Neisseria include one or more selected from the group consisting of Neisseria mucosa and Neisseria flavescens, and the bacteria of the genus Haemophilus include Haemophilus parainfluenzae, satisfying at least any one of the above.

4. The agent according to claim 1 or 2, further containing a nitrate ion supply compound.

5. The agent according to claim 1 or 2, which is a caries prevention / amelioration agent or a periodontal disease prevention / amelioration agent.

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