Oral nitrate-reducing bacteria growth enhancer

Abstract

To provide an oral nitrate-reducing bacteria growth enhancer that allows selective growth of nitrate-reducing bacteria in the oral cavity and controls the balance of oral bacterial flora.SOLUTION: An oral nitrate-reducing bacteria growth enhancer comprises a nitrate ion-supplying compound and a compound having a sterol skeleton, preferably one represented by formula (1).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 of the bacteria that cause diseases but also of the 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 (NO 3 - ) to nitrite ions (NO 2 - ) or nitric oxide (NO). As mechanisms by which nitrate-reducing bacteria, i.e., bacteria having nitrate-reducing ability, bring benefits to the host human, there are known the blood pressure-lowering action of nitric oxide, the antibacterial action against periodontal pathogenic bacteria, and the action of preventing dental demineralization by suppressing a decrease in salivary pH. Therefore, selectively growing nitrate-reducing bacteria in the oral cavity and controlling the balance of the oral microbiota are useful in preventing dental caries and periodontal disease.

[0003] Since nitrate-reducing bacteria use nitrate as a major 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, there is a desire for a means of growing nitrate-reducing bacteria even under conditions of low nitrate concentration in the oral cavity. Thus, an attempt was made to search for an agent for promoting the growth of nitrate-reducing bacteria among additives (quasi-drug additives, food additives).

[0004] As techniques for controlling the microbiota balance, the following have been proposed. Patent Document 1 describes an oral Streptococcus mutans growth promoter containing a nitrate ion supply compound and a sugar alcohol as active ingredients. Patent Document 2 describes an oral care composition containing a water-soluble nitrate, a basic amino acid, a water-soluble alkali metal polyphosphate, and water. Patent Document 3 describes an intestinal flora composition regulator containing γ-oryzanol and biocompatible particles encapsulating γ-oryzanol, and sterols are exemplified as the biocompatible particles.

Prior Art Documents

Patent Documents

[0005]

Patent Document 1

Patent Document 2

Patent Document 3

Summary of the Invention

Problems to be Solved by the Invention

[0006] However, the effect of selectively growing nitrate-reducing bacteria by a nitrate ion supply compound and a compound having a sterol skeleton is not known.

[0007] An object of the present invention is to provide an oral nitrate-reducing bacteria growth promoter that selectively grows nitrate-reducing bacteria in the oral cavity and controls the oral bacterial flora balance.

Means for Solving the Problems

[0008] The present invention provides the following [1] to

[11] . 〔1〕An oral nitrate-reducing bacteria growth promoter containing (A) component: a nitrate ion supply compound and (B) component: a compound having a sterol skeleton. 〔2〕The agent according to [1], wherein the (B) component is a compound represented by the formula (1).

Chemical Formula

Chemical formula

Advantages 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 the balance of the oral bacterial flora can be controlled by nitrite and nitric oxide, so that dental caries and periodontal disease can be prevented or improved.

Mode for Carrying Out the Invention

[0010] [1. Oral nitrate-reducing bacteria growth agent] The oral nitrate-reducing bacteria growth agent contains the following components (A) and (B).

[0011] [Component (A)] Component (A) is a nitrate ion supply compound. Component (A) can supply nitrate ions utilized by nitrate-reducing bacteria.

[0012] As the nitrate ion supply compound, any compound that can supply nitrate ions (NO 3 - ) may be used. Examples include nitrate esters and nitrates, and nitrates are preferred. Examples of the salts of nitrates include counter ions of nitrate ions such as alkali metal salts (e.g., sodium salts, potassium salts), alkaline earth metal salts (e.g., calcium salts), and ammonium salts. The nitrates are preferably potassium nitrate, sodium nitrate, calcium nitrate, and ammonium nitrate, more preferably potassium nitrate, sodium nitrate, and ammonium nitrate, and still more preferably potassium nitrate.

[0013] The nitrate ion supply compound may be any of a natural raw material (natural) containing the nitrate ion supply compound itself, an extract or purified product obtained from the above natural raw material, and a chemically synthesized product containing the nitrate ion supply compound. 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 thereof (e.g., vegetable extracts, powders). Component (A) may be a single nitrate ion supply compound or a combination of two or more.

[0014] [Component (B)] Component (B) is a compound having a sterol skeleton (hereinafter also referred to as "sterols"), and is preferably a compound represented by the following formula (1). By containing Component (B), while suppressing the intraoral concentration of Component (A), the agent of the present invention can be imparted with an effect of promoting the growth of nitrate-reducing bacteria. [Chemical formula]

[0015] In formula (1), R 1 represents a hydrogen atom or a group represented by the following formula (2). [Chemical formula]

[0016] In formula (2), R 9 represents an alkyl group, or an alkenyl group having an aromatic ring having an alkoxy group and a hydroxyl group. The number of carbon atoms of the alkyl group is preferably 1 to 7, more preferably 1 to 5, and still more preferably 1 to 3. Each group may be linear or branched, but a linear chain is preferred. The alkyl group is preferably a propyl group, and more preferably an n-propyl group. The number of carbon atoms of the alkenyl group is preferably 1 to 7, more preferably 1 to 5, and still more preferably 1 to 3. The alkenyl group may be linear or branched, but a linear chain is preferred. Also, the number of carbon atoms of the alkoxy group is preferably 1 to 5, more preferably 2 to 3, and still more preferably 2. Examples of the alkenyl group having an aromatic ring having an alkoxy group and a hydroxyl group include a 4-hydroxy-3-methoxyphenyl-2-ethenyl group.

[0017] In formula (1), R 2 , R 3 and R 5preferably each independently represents a hydrogen atom or an alkyl group, more preferably commonly represents a hydrogen atom or an alkyl group. The number of carbon atoms of the alkyl group is preferably 1 to 7, more preferably 1 to 5, and even more preferably 1 to 3. The alkyl group is preferably a methyl group, an ethyl group, or a propyl group, and more preferably a methyl group.

[0018] In formula (1), R 4 , and R 7 preferably each independently represents a hydrogen atom or a hydroxyl group.

[0019] In formula (1), R 6 represents an alkyl group which may have a carboxyl group or an alkenyl group. The number of carbon atoms of the alkyl group and the alkenyl group is preferably 1 to 10, more preferably 1 to 9, and even more preferably 2 to 7. The alkyl group may be linear or branched, preferably linear when having a carboxyl group, and preferably branched when not having a carboxyl group. The alkenyl group may be linear or branched, but is preferably branched. As the alkyl group having a carboxyl group, a 2-carboxyethyl group is preferred. As the alkyl group not having a carboxyl group, a 4-methylpentyl group, a 3-ethyl-4-methyl-pentyl group, a 3,4-dimethylpentyl group, and a 3-methylidene-4-methylpentyl group are preferred. As the alkenyl group, a 4-methyl-3-pentenyl group, a 3,4-dimethyl-1-pentenyl group, a 3-ethyl-4-methyl-1-pentenyl group, and a 2,4-dimethyl-3-pentenyl group are preferred.

[0020] R 8 is an alkyl group, and the carbon atom constituting the alkyl group may combine with the carbon atom C 1 to form a ring. The number of carbon atoms of the alkyl group is preferably 1 to 5, more preferably 1 to 3, and even more preferably 1. The ring formed by the carbon atom constituting the alkyl group combining with the carbon atom C 1 includes, for example, a three-membered ring, a four-membered ring, a five-membered ring, and a six-membered ring, and is preferably a three-membered ring. Therefore, R 8 is a methyl group or a carbon atom, and the carbon atom is the carbon atom C1 It is more preferable to form a three-membered ring by combining with

[0021] In the formula, the solid and dotted double lines indicate a single bond or a double bond.

[0022] Component (B) is preferably at least one selected from the following group of compounds; phytosterol, cholesterol: formula (3), lanosterol: formula (4), cholesteryl butyrate: formula (5), γ-oryzanol, and dihydrocholesterol: formula (6). Among these, phytosterol, cholesterol, lanosterol, cholesteryl butyrate, β-sitosterol, γ-oryzanol are more preferable, and phytosterol is even more preferable. [Chemical formula]

[0023] Phytosterol is the main component (usually 85.0% by mass or more) of plant sterols and contains one or more of various sterols such as sitosterol (e.g., β-sitosterol: formula (7)), stigmasterol: formula (8), campesterol: formula (9), and brassicasterol: formula (10), etc., which are represented by the following structural formula. [Chemical formula]

[0024] γ-Oryzanol is a general term for esters in which ferulic acid and sterols are condensed and contained in rice bran. For example, it contains one or more selected from cycloartenol ferulate: formula (11), 24-methylenecycloartanol ferulate: formula (12), campesterol ferulate: formula (13), β-sitosterol ferulate: formula (14), and cyclobranol ferulate: formula (15), which are represented by the following structural formula. [Chemical formula]

[0025] The compound represented by the general formula (1) may be any of the natural raw material (natural) itself containing the compound, an extract or purified product obtained from the above raw material, a chemical synthetic product containing the compound represented by the general formula (1), or a product purified to the compound. The component (B) may be a single compound represented by the general formula (1) or a combination of two or more thereof.

[0026] [Effective amount] -(Component (A))- The effective amount (mass fraction) of the component (A) in the oral nitrate-reducing bacteria growth agent is preferably 50 ppm or more, more preferably 100 ppm or more, and still more preferably 200 ppm or more. Thereby, nitrate ions can be sufficiently provided to the nitrate-reducing bacteria. The upper limit is preferably 5000 ppm or less, more preferably 1000 ppm or less, and still more preferably 800 ppm or less. Thereby, the oral nitrate concentration can be suppressed within an appropriate range. Therefore, the effective amount of the component (A) is preferably 50 to 5000 ppm, more preferably 100 to 1000 ppm, and still more preferably 200 to 800 ppm. The above-mentioned "effective amount" indicates the effective amount of the component (A) in the saliva secreted into the oral cavity assuming that the saliva secretion rate is 1 mL / min. The same applies to the effective amount of the following component (B).

[0027] -(Component (B))- The effective amount (mass fraction) of the component (B) in the total amount of the oral nitrate-reducing bacteria growth agent is preferably 5 ppm or more, more preferably 7 ppm or more, and still more preferably 9 ppm or more. Thereby, the oral nitrate-reducing bacteria growth effect can be exhibited. The upper limit is preferably 10000 ppm or less, more preferably 8000 ppm or less, and still more preferably 6000 ppm or less. Thereby, the oral nitrate-reducing bacteria can be moderately grown. Therefore, the effective amount of the component (B) is preferably 5 to 10000 ppm, more preferably 7 to 8000 ppm, and still more preferably 9 to 6000 ppm.

[0028] -Effective amount ratio of component (B) to component (A)- (A) The effective amount ratio of component (B) to component (A) is preferably 0.005 or more, more preferably 0.01 or more, still more preferably 0.015 or more, or 0.02 or more. The upper limit is preferably 50 or less, more preferably 30 or less, still more preferably 20 or less, or 10 or less. Therefore, the effective amount ratio of component (B) to component (A) is preferably 0.005 to 50, more preferably 0.01 to 30, still more preferably 0.015 to 20 or 0.02 to 10. By the effective amount ratio of component (B) to component (A) satisfying the above numerical range, the effects of the present invention can be more preferably exerted.

[0029] [2. Nitrate-reducing bacteria growth effect] The oral nitrate-reducing bacteria growth agent 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 state the number of nitrate-reducing bacteria in the oral cavity and / or the ratio of the number of nitrate-reducing bacteria to the total number of bacteria in the oral cavity.

[0030] -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 Haemophilus, genus Neisseria, genus Veillonella, genus Granulicatella, genus Lautropia, genus Rothia, genus Actinomyces, genus Leptotrichia, and genus Scardovia can be mentioned.

[0031] Examples of the bacteria of the genus Haemophilus include Haemophilus parainfluenzae. Examples of bacteria of the genus Neisseria include, for example, Neisseria mucosa, Neisseria flavescens, Neisseria subflava, Neisseria perflava, Neisseria sicca, and Neisseria flava. Examples of bacteria of the genus Veillonella include, for example, Veillonella parvula. Examples of bacteria of the genus Granullicatella include, for example, Granullicatella adiacens. Examples of bacteria of the genus Lautropia include, for example, Lautropia mirabilis. Examples of bacteria of the genus Rothia include, for example, Rothia mucilaginosa, Rothia dentocariosa, and Rothia aeria. Examples of bacteria of the genus Actinomyces include, for example, Actinomyces viscosus and Actinomyces naeslundii. Examples of bacteria of the genus Leptotrichia include, for example, Leptotrichia buccalis. Examples of bacteria of the genus Schaalia include, for example, Schaalia odontolytica.

[0032] Among these, bacteria of the genus Haemophilus, Neisseria, and Veillonella are preferred, and Haemophilus parainfluenzae, Neisseria mucosa, and Veillonella parvula are more preferred.

[0033] Confirmation of the effect of promoting the growth of nitrate-reducing bacteria in the oral cavity can be achieved, for example, by determining the ratio (nitrate-reducing bacteria ratio) of the total number of bacteria in the oral cavity to the total number of bacteria of Haemophilus spp., Neisseria spp., Veillonella spp., Granulicatella spp., Rothia spp., Actinomyces spp., Leptotrichia spp., and Scardovia spp. This ratio is usually 30% or more, preferably 35% or more, more preferably 50% or more, and even more preferably 60% or more. The measurement conditions for the number of each type of bacteria can be carried out according to the conditions described in the examples below.

[0034] -Other effects- The combination of the above components (A) and (B) can exert effects associated with the effect of promoting the growth of nitrate-reducing bacteria. For example, it can improve the oral flora, prevent and improve dental caries and / or periodontal disease, increase nitric oxide, improve and enhance motor function, and prevent and improve hypertension.

[0035] -Administration method and target- Examples of the administration method of the agent include oral administration (e.g., intraoral administration, sublingual administration), parenteral administration (e.g., transdermal administration, intravenous administration, intramuscular administration, subcutaneous administration, nasal administration, pulmonary administration). Among these, an administration form with less invasiveness is preferred, and oral administration (oral ingestion) is more preferred.

[0036] The administration target can be any animal including humans, usually humans. The administration target can be a healthy person, but it can also be a person infected with a bacterial infection in the oral cavity (e.g., 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.

[0037] [3. Use as pharmaceuticals, quasi-drugs, cosmetics, and foods] The agent for promoting the growth of nitrate-reducing bacteria in the oral cavity can be used as pharmaceuticals, quasi-drugs, cosmetics, and foods. Examples of the dosage form include liquid agents, spray agents, solid agents, semi-solid agents, liquid agents, powder agents, and granule agents, which can be determined according to various uses.

[0038] [4. Oral compositions] The above components (A) and (B) can be used as an oral composition. Thereby, the effect of promoting the growth of nitrate-reducing bacteria in the oral cavity can be efficiently exerted.

[0039] [Content of components (A) and (B) in the oral composition] -(Component (A)- When the agent of the present invention is an oral composition, the content of component (A) in the composition is as follows.

[0040] When the oral composition is a liquid preparation, the content of component (A) in the composition is preferably 10 mg / mL or more, more preferably 20 mg / mL or more. Thereby, nitrate ions can be sufficiently provided to nitrate-reducing bacteria. The upper limit is preferably 1000 mg / mL or less, more preferably 200 mg / mL or less. Thereby, the nitrate concentration in the oral cavity can be suppressed within an appropriate range. Therefore, the content of component (A) in the composition is preferably 10 to 1000 mg / mL, more preferably 20 to 200 mg / mL.

[0041] When the oral composition is a solid such as a tablet, the content of component (A) in the composition is preferably 10 mg / g or more, more preferably 20 mg / g or more. Thereby, nitrate ions can be sufficiently provided to nitrate-reducing bacteria. The upper limit is preferably 1000 mg / g or less, more preferably 200 mg / g or less. Thereby, the nitrate concentration in the oral cavity can be suppressed within an appropriate range. Therefore, the content of component (A) in the composition is preferably 10 to 1000 mg / g, more preferably 20 to 200 mg / g.

[0042] (A) The daily intake of the component is not particularly limited, but when targeting humans, it is usually 20 to 500 mg / day, preferably 20 to 300 mg / day, more preferably 30 to 200 mg / day, and even more preferably 60 to 200 mg / day.

[0043] -(Component (B)- When the agent of the present invention is an oral composition, the content of component (B) in the composition is as follows.

[0044] When the oral composition is a liquid preparation, the content of component (B) in the composition is preferably 10 mg / mL or more, more preferably 100 mg / mL or more. The upper limit is preferably 3000 mg / mL or less, more preferably 500 mg / mL or less. Therefore, the content of component (B) in the composition is preferably 10 to 3000 mg / mL, more preferably 100 to 500 mg / mL.

[0045] When the oral composition is a solid such as a tablet, the content of component (B) in the composition is preferably 10 mg / g or more, more preferably 100 mg / g or more. The upper limit is preferably 3000 mg / g or less, more preferably 500 mg / g or less. Therefore, the content of component (B) in the composition is preferably 10 to 3000 mg / g, more preferably 100 to 500 mg / g.

[0046] The daily intake of component (B) is not particularly limited, but when targeting humans, it is usually 10 to 4000 mg / day, preferably 50 to 3000 mg / day, more preferably 100 to 1000 mg / day.

[0047] -Mass ratio of component (B) to component (A) in the oral composition- The mass ratio of component (B) to component (A) is preferably 0.005 or more, more preferably 0.01 or more, still more preferably 0.015 or more, or 0.02 or more. The upper limit is preferably 50 or less, more preferably 30 or less, still more preferably 20 or less, or 10 or less. Therefore, the mass ratio of component (B) to component (A) is preferably 0.005 to 50, more preferably 0.01 to 30, still more preferably 0.015 to 20 or 0.02 to 10. By the mass ratio of component (B) to component (A) satisfying the above numerical range, the effects of the present invention can be more preferably exerted.

[0048] As used herein, an oral composition means a composition applied to the oral cavity, regardless of whether it is a medicine, quasi-drug, food, or cosmetic. For example, dentifrices (e.g., paste dentifrices, gel dentifrices, moisturizing dentifrices, liquid dentifrices), mouthwashes, tongue cleaners, oral sprays, tablets, oral films, gums, oral refreshers, gargle tablets, oral pastes, gels, and ointments can be mentioned. Examples of foods (food compositions) include food compositions for uses such as health foods, functional foods, health foods, dietary supplements, nutritional supplements, foods for specified health uses, foods with nutritional functions, medical foods, foods for patients, foods for infants, foods for nursing care, and foods for the elderly, and processed products thereof (e.g., drinks). Among these, tablets, drinks, or oral films are preferred.

[0049] [Dosage form] Examples of dosage forms include 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 can be appropriately selected according to the use.

[0050] [Optional components] The oral composition may contain components other than components (A) and (B). Examples of other components include bactericides, preservatives, medicinal ingredients, surfactants, abrasives, wetting agents, binders, buffers, solubilizing agents, 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, seasonings, food raw materials (including food additives), 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, food compositions, and cosmetics, and / or dosage forms, administration methods, etc., and may be one type or a combination of two or more types.

[0051] -Bactericide- Examples of the bactericide include cetylpyridinium chloride, benzalkonium chloride, benzethonium chloride, alkyldiaminoethyl glycine hydrochloride, chlorhexidine or its salts, triclosan, isopropylmethylphenol, hinokitiol, chlorhexidine gluconate, decalinium chloride, iodine, potassium iodide, sulfamethoxazole, sodium sulfamethoxazole, sulfisoxazole, and sodium sulfisomidine.

[0052] -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.

[0053] In the present invention, it is more suitable in terms of the growth of nitrate-reducing bacteria not to contain the above bactericide and / or preservative (each content is 0%), but 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.

[0054] -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, glycyrrhetinate (e.g., dipotassium glycyrrhizinate), allantoin chlorhydroxyaluminum, azulene, and dihydrocholesterol; metal salts such as zinc salts, copper salts, and tin salts; tartar inhibitors 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 hydroxyethyl cellulose 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; tartar inhibitors; amino acids such as alanine, glycine, and proline; plant extracts such as thyme, saffron, cloves, and witch hazel; caropeptide; polyvinylpyrrolidone, and the like. Other examples include decongestants, anti-inflammatory agents, astringents, antihistamines, vitamins, amino acids, bactericides, local anesthetics, components having a flora-improving 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, dipotassium glycyrrhetinate, ammonium glycyrrhetinate, glycyrrhetinic acid, 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 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 amino acids include potassium L-aspartate, magnesium L-aspartate, aminoethylsulfonic acid, and sodium chondroitin sulfate. Examples of local anesthetics include lidocaine, lidocaine hydrochloride, dibucaine hydrochloride, and chlorobutanol. Each medicinal ingredient may be used alone or in combination of two or more. The content of the medicinal ingredient can be appropriately set to an effective amount according to conventional methods.

[0055] -Surfactant- Examples of surfactants include nonionic surfactants, anionic surfactants, and amphoteric surfactants.

[0056] Examples of nonionic surfactants include polyoxyethylene alkyl ethers, polyoxyethylene hydrogenated castor oil, sorbitan fatty acid esters, polyoxyethylene sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monostearate), alkylolamides, polyoxyethylene fatty acid esters, polyglycerin fatty acid esters, sucrose fatty acid esters (e.g., maltose fatty acid esters), sugar alcohol fatty acid esters (e.g., maltitol fatty acid esters, lactitol fatty acid esters), fatty acid diethanolamides (e.g., lauric acid mono- or diethanolamide), polyoxyethylene polyoxypropylene copolymers, polyoxyethylene polyoxypropylene fatty acid esters, glycerin fatty acid esters, polyoxyethylene glycerin fatty acid esters, polyoxyethylene glycol fatty acid esters, polyoxyethylene lanolin and lanolin alcohol, polyoxyethylene alkylamines and fatty acid amides, polyoxyethylene alkyl phenyl formaldehyde condensates, polyoxyethylene polyoxypropylene alkyl ethers, and polyoxyethylene alkyl phenyl ethers. Among these, polyoxyethylene alkyl ethers and polyoxyethylene hydrogenated castor oil are preferred, and polyoxyethylene hydrogenated castor oil is more preferred.

[0057] 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 straight-chain or branched-chain, 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 salts, potassium salts, calcium salts, magnesium salts, and ammonium salts; organic base salts such as triethylammonium salts, triethanolammonium salts, pyridinium salts, and diisopropylammonium salts; and basic amino acid salts such as arginine salts. Among them, inorganic base salts are preferred, alkali metal salts (for example, sodium salts, potassium salts) or ammonium salts are more preferred, and sodium salts are even more preferred. Examples of alkyl sulfates include, for example, myristyl sulfate. Examples of acyl amino acid salts include, for example, 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 salts and N-cocoyl methyl taurine salts. Examples of anionic surfactants also include sodium hydrogenated coconut fatty acid monoglyceride monosulfate and sodium lauryl sulfacetate.

[0058] Examples of the amphoteric surfactant include betaine-type amphoteric surfactants such as alkyldimethylaminoacetic acid betaine (e.g., lauryldimethylaminoacetic acid betaine), fatty acid amidopropyldimethylaminoacetic acid betaine; imidazoline-type amphoteric surfactants such as N-fatty acyl-N-carboxymethyl-N-hydroxyethyl ethylenediamine salt, coconut oil fatty acid imidazolinium betaine; and alkyl betaines such as lauryldimethylaminoacetic acid betaine.

[0059] When the surfactant is included, 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 agent.

[0060] - Abrasive - As the abrasive, for example, either an inorganic abrasive or an organic abrasive may be used. Examples of the inorganic abrasive 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, trimagnesium 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 the organic abrasive include polymethyl methacrylate and synthetic resin-based abrasives. Among these, abrasive silica and calcium phosphate-based compounds are preferred, and anhydrous silicic acid is more preferred. The amount of the abrasive is preferably 50% by mass or less, more preferably 8 to 50% by mass based on the whole composition.

[0061] - 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, and 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 Pharmaceutical Excipients Standard. The content of the wetting agent is usually 40% by mass or less, preferably 1 to 30% by mass, based on the whole composition.

[0062] -Binder- Examples of the binder include any conventionally known and suitable organic binders, 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), and synthetic water-soluble polymers (e.g., sodium polyacrylate, carboxyvinyl polymer, polyvinyl pyrrolidone, polyvinyl alcohol, propylene glycol alginate). Furthermore, 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.

[0063] -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.

[0064] - Solubilizing agent - Examples of solubilizing agents include propylene glycol, polyethylene glycol, and combinations of two or more of these.

[0065] - Isotonic agent - Examples of isotonic agents include sodium chloride, potassium chloride, glycerin, and combinations of two or more of these.

[0066] - 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.

[0067] - Chelating agent - Examples of chelating agents include sodium edetate, sodium citrate, and combinations of these.

[0068] - Humectant - Examples of humectants include glycerin, concentrated glycerin, sugar alcohols (e.g., sorbitol, xylitol, maltitol, mannitol, reduced starch syrup, reduced palatinose, erythritol, lactitol, isomalt), and combinations of two or more of these.

[0069] - Flavoring agent - Examples of flavoring agents include sweeteners (e.g., sodium saccharin, dipotassium glycyrrhizinate, aspartame, stevia, stevioside, paramethoxycinnamic aldehyde, neohesperidin dihydrochalcone, perillartine, aspartylphenylalanine methyl ester, thaumatin, acesulfame potassium, sucralose, maltitol, sorbitol, mannitol, reduced maltose, reduced palatinose, xylitol, erythritol, lactitol, etc., artificial sweeteners); 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., fragrance components contained in the above natural essential oils; ethyl acetate, ethyl butyrate, isoamyl acetate, hexanal, hexanal, methyl anthranilate, ethyl methylphenylglycidate, benzaldehyde, vanilla, ethyl vanillin, furaneol, N-ethyl-p-menthane-3-carboxamide, menthyl lactate, ethylene glycol-l-menthyl carbonate, etc., fragrance components; and various compounded flavors such as mint, fruit, herb, etc., formed by combining some 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.

[0070] - 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.

[0071] - 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 alcohol include glycerin; glycols such as ethylene glycol, propylene glycol, dipropylene glycol, butylene glycol, polyethylene glycol (PEG); and reduced starch saccharides. As the polyethylene glycol, 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 Raw Material Specifications for Quasi-Drugs 2006. The amount of the humectant is usually 40% by mass or less, preferably 1 to 30% by mass, based on the whole composition.

[0072] - Colorant - Examples of the colorant include natural colorants such as safflower red pigment, gardenia yellow pigment, gardenia blue pigment, perilla pigment, red kojic pigment, red cabbage pigment, carrot pigment, hibiscus pigment, cocoa pigment, spirulina blue pigment, tamarind pigment, etc., legal colorants 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.

[0073] -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 (e.g., potassium salt, sodium salt, and ammonium salt), and 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 agent after addition is 5 to 9, preferably 6 to 8.5. In this specification, the pH value usually refers to the value 3 minutes after the start of measurement at 25°C. The pH value can be measured, for example, using a pH meter (model number Hm-30S) manufactured by Toa Denpa Kogyo Co., Ltd.

[0074] -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.

[0075] -Excipient- Examples of excipients include celluloses 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, 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 particulate silicon dioxide), titanium oxide, and aluminum hydroxide gel; and combinations of two or more of these.

[0076] - 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.

[0077] - Binder - Examples of binders include hydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose, ethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, gelatin, dextrin, starch, pregelatinized starch, and combinations of two or more of these.

[0078] - 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.

[0079] -Other optional components- Examples of the optional components other than the 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 hinder the effects of the present invention.

[0080] [5. Manufacturing method] The method for manufacturing an oral nitrate-reducing bacteria 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, after preparing the components that dissolve in a solvent, the other insoluble components are mixed, and defoaming (e.g., under reduced pressure) is performed as necessary. Another example is a method in which the active 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). As the container for the oral agent, for example, a laminated tube can be mentioned, and as the material, resins such as polyethylene, polypropylene, polyethylene terephthalate, and nylon can be used. In the case of a spray agent, a container equipped with a spraying means (e.g., a trigger type, pump type, aerosol type container) can be selected. The obtained dentifrice can be contained in a container to make a product. The container is not particularly limited in shape and material, and a container usually used for an oral composition can be used.

[0081] [6. Usage method] The method of using the oral nitrate-reducing bacteria growth agent may be, for example, to administer the agent to the application site. The number of administrations per day is not particularly limited, but 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

[0082] Hereinafter, the present invention will be described with reference to examples. The following examples do not limit the present invention.

[0083] (1) Preparation of a multi-species biofilm using human saliva To 20 mL of a liquid medium (Table 1), 16 μL of a 0.4 ppm vitamin K1 solution (manufactured by FUJIFILM Wako Pure Chemical Corporation), 400 μL of a 350,000 ppm sucrose aqueous solution (manufactured by FUJIFILM Wako Pure Chemical Corporation), and 800 μL of resting saliva were added to prepare a medium (medium (1)). To each well of a 24-well plate (manufactured by Corning), a control (0.75 mL of medium (1) and 0.75 mL of a 10 mM potassium nitrate aqueous solution; Comparative Examples 1 to 3), a seeds·potassium nitrate addition group (0.75 mL of medium (1), 0.75 mL of a 10 mM potassium nitrate aqueous solution, 1% of the active ingredient ((B) component) with respect to medium (1); Examples 1 to 9), and a potassium nitrate non-addition group (1.5 mL of medium (1), 1% of the active ingredient ((B) component) with respect to medium (1); Comparative Examples 4 to 6) were added and mixed respectively. A test plate equipped with a HAP plate was inserted into the 24-well plate, and anaerobic culture was performed at 37°C. After 8 hours, the HAP plate was washed with a washing solution (Table 2), and medium (1), the potassium nitrate aqueous solution, and seeds (component (B)) were added as described above, and anaerobic culture was performed at 37°C. After 16 hours, the HAP plate was washed again with the washing solution. Thereafter, the HAP plate was washed and the medium was exchanged for one week (excluding Saturdays and Sundays) to form a biofilm on the HAP plate. After culture, the HAP plate was removed with tweezers and transferred to a test tube containing 2 mL of PBS (manufactured by gibco), and ultrasonic treatment (manufactured by BRANSON) was performed 10 times for 1 second. The bacterial suspension was transferred to a 2 mL tube, centrifuged at 16,400×g for 5 minutes, and the supernatant was removed and the pellet was collected.

[0084]

Table 1

[0085] [Footnote of Table 1] ※1 The liquid medium was prepared by adjusting the composition in Table 1 to pH 7.0 with 1N NaOH, making up to 1 L with distilled water, and subjecting it to autoclave treatment (121°C, 40 minutes). ※2 The hemin solution was prepared according to the following procedure. 0.25 g of Hemin (manufactured by Fujifilm Wako Pure Chemical Corporation) was placed in a 500 mL beaker, and 5 mL of 1N NaOH was added and mixed well. Approximately 400 mL of distilled water was added, and the mixture was stirred with a stirrer. It was adjusted to pH 7.0 with 1N HCl, made up to 500 mL, and autoclaved (121 °C, 20 minutes).

[0086]

Table 2

[0087] [Footnote of Table 2] ※ The HAP plate cleaning solution was prepared by adjusting the composition of Table 1 to pH 7.3 with 1N NaOH, making up to 1 L with distilled water, and autoclaving (121 °C, 40 minutes).

[0088] (2) DNA extraction DNA extraction was performed on the pellet recovered in (1) using the Nexttec 1-Step DNA Isolation Kit (manufactured by TOHO).

[0089] (3) Bacterial flora analysis Using the next-generation sequencer MiSeq (manufactured by Illumina), the bacterial flora was analyzed as follows. The amplification primers for the V1-V2 region of the 16S rRNA gene shown in Table 3 (forward primer: 27Fmod consisting of the nucleotide sequence represented by SEQ ID NO: 1, reverse primer: 338R consisting of the nucleotide sequence represented by SEQ ID NO: 2) were used to amplify the 16S rRNA gene using Ex Taq polymerase (manufactured by Takara Bio Inc.) by performing PCR under the composition shown in Table 4 and the conditions shown in Table 5. The obtained PCR product was subjected to electrophoresis to confirm the amplification of the sequence length of the target region. DNA purification was performed using AM Pure XP (manufactured by Beckman Coulter) according to a predetermined procedure. The purified samples were measured for concentration using the Quant-iT® PicoGreen® dsDNA Assay Kit (manufactured by Thermo Fisher Scientific), and libraries were prepared by mixing the samples so that each sample had the same amount of DNA. Library purification was performed using the MinElute PCR Purification Kit (manufactured by QIAGEN) according to a predetermined procedure. The concentration of the library was measured by real-time PCR using the KAPA Library Quantification Kit (manufactured by Kapa Biosystems). The sequence length of the library was confirmed using a Bioanalyzer (manufactured by Agilent). Acquisition of sequence information was performed using the MiSeq Reagent Kit v3 (Illumina) and the next-generation sequencer MiSeq (Illumina) according to a predetermined procedure.

[0090]

Table 3

[0091]

Table 4

[0092]

Table 5

[0093] The obtained sequences were compared with the Forward primer, the sequence containing both the Forward primer and the Reverse primer, the sequence with a Quality value indicating the reliability of the sequence being 25 or more, and the gene sequences in the database, and filtering was performed on the condition that the alignment length was 90% or more. Only the sequences that passed the filtering were used for subsequent analysis. For each sample, 10,000 sequences were randomly extracted to unify the number of data. Sequences with a homology of 97% or more were grouped as one Operational Taxonomic Unit (OTU). The representative sequence of each OTU was compared with four public databases (RDP, CORE, NCBI, HOMD) to identify bacterial species. For the analysis at the species level, only bacterial species with a homology of 97% or more were adopted, and the rest were regarded as Undefined.

[0094] (2) Using the sample obtained by diluting the extracted DNA 100-fold as a template, quantitative PCR was performed using SsoAdvanced Universal Probes Supermix (manufactured by BIO-RAD) (Table 6). The probes and primers shown in Table 7 were used. Also, the reaction conditions were carried out as shown in Table 8, and for the calibration curve, a plasmid with a known concentration (10 8 copies / μL) was used and diluted stepwise up to 10 7 times. From the DNA concentration of the calibration curve and the number of cycles (Ct value) in the linear part of the curve, a calibration curve was created, and the total number of bacteria (copies / μL) in each sample was calculated.

[0095]

Table 6

[0096]

Table 7

[0097]

Table 8

[0098] (5) Data analysis For each evaluation sample, taking the ratio of nitrate-reducing bacteria in the bacterial flora measured by the bacterial flora analysis in (3) (total ratio of Haemophilus, Neisseria, Veillonella, Granullicatella, Lautropia, Rothia, Actinomyces, Leptotrichia, Schaalia, which are known as nitrate-reducing bacteria more common in healthy individuals) as a (%), and the total number of bacteria (copies / μL) as b, the number of nitrate-reducing bacteria (copies / μL) was calculated using the following formula. The growth effect on nitrate-reducing bacteria was evaluated from the ratio and number of nitrate-reducing bacteria (Tables 9 and 10). Number of nitrate-reducing bacteria (copies / μL) = b × a / 100

[0099] [Potassium nitrate and seeds used] Potassium nitrate (manufactured by FUJIFILM Wako Pure Chemical Corporation) Phytosterol (raw material name: Phytosterol-FKP, manufactured by Tama Seikagaku Corporation) Cholesterol (manufactured by Kanto Chemical Co., Inc.) Lanosterol (manufactured by Tokyo Chemical Industry Co., Ltd.) Cholesteryl butyrate (manufactured by Tokyo Chemical Industry Co., Ltd.) β-Sitosterol (manufactured by FUJIFILM Wako Pure Chemical Corporation) γ-Oryzanol (raw material name: ORYZA GAMMAX, manufactured by Oryza Oil & Fat Chemical Co., Ltd.) Dihydrocholesterol (manufactured by Tokyo Chemical Industry Co., Ltd.)

[0100]

Table 9

[0101]

Table 10

[0102] In the comparative examples using component (A) or component (B), the ratio of nitrate-reducing bacteria was less than 30% in both cases. On the other hand, in the examples using the agent of the present invention combining component (A) and component (B), the ratio of nitrate-reducing bacteria in all cases exceeded 30%.

[0103] For example, the ratio of nitrate-reducing bacteria in Example 2 far exceeded the numerical value obtained by adding the ratio of nitrate-reducing bacteria by the sole use of 500 ppm of component (A) in Comparative Example 2 and the ratio of nitrate-reducing bacteria by the sole use of 1000 ppm of component (B) in Comparative Example 5. From this, it is shown that by combining component (A) and component (B) in the present invention, a synergistic effect is exerted, and an unexpected and remarkable nitrate-reducing bacteria growth effect is exerted.

[0104] These results indicate that the agent of the present invention can selectively grow nitrate-reducing bacteria in the oral cavity and control the oral bacterial flora balance.

Claims

1. Component (A): A nitrate ion supply compound, and Component (B): A compound having a sterol skeleton An oral nitrate-reducing bacteria growth promoter containing the same.

2. The agent according to claim 1, wherein component (B) is a compound represented by formula (1). 【Chemical 1】 (In formula (1), R １ represents a hydrogen atom or a group represented by the following formula (2). R ２ , R ３ and R ５ each independently represent a hydrogen atom or an alkyl group having 1 to 7 carbon atoms. R ４ , and R ７ each independently represent a hydrogen atom or a hydroxyl group. R ６ represents an alkyl group having 1 to 10 carbon atoms which may have a carboxyl group, or an alkenyl group having 1 to 10 carbon atoms. R ８ is an alkyl group, and the carbon atoms constituting the alkyl group may combine with the carbon atom C １ to form a ring. In the formula, the solid and dotted double lines represent a single bond or a double bond.) [Chemical 2] (In formula (2), R ９ represents an alkyl group or an alkoxy group having 1 to 7 carbon atoms, and an alkenyl group having 1 to 7 carbon atoms and an aromatic ring having a hydroxyl group.)

3. The agent according to claim 1 or 2, wherein component (A) contains one or more selected from the group consisting of potassium nitrate, sodium nitrate, and ammonium nitrate.

4. The agent according to claim 1 or 2, wherein component (B) contains one or more selected from the group consisting of phytosterol, cholesterol, lanosterol, cholesteryl butyrate, β-sitosterol, γ-oryzanol, and dihydrocholesterol.

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

6. The bacteria of the genus Haemophilus contain Haemophilus parainfluenzae, The bacteria of the genus Neisseria contain one or more selected from the group consisting of Neisseria mucosa, Neisseria flavescens, Neisseria subflava, Neisseria perflava, Neisseria sicca, and Neisseria flava, The bacteria of the genus Veillonella contain Veillonella parvula, The bacteria of the genus Granulicatella contain Granulicatella adiacens, The bacteria of the genus Rothia contain Rothia mucilaginosa, The bacteria of the genus Rothia contain one or more selected from the group consisting of Rothia dentocariosas, and Rothia aeria, The bacteria of the genus Actinomyces contain one or more selected from the group consisting of Actinomyces viscosus, and Actinomyces naeslundii, The bacteria of the genus Leptotrichia contain Leptotrichia buccalis, and The bacteria of the genus Scardovia contain Scardovia odontolytica, The agent according to claim 5, satisfying at least any one of the above.

7. The effective amount of component (A) is 50 to 5000 ppm, or The agent according to claim 1 or 2, wherein the effective amount of component (B) is 5 to 10000 ppm.

8. The agent according to claim 1 or 2, wherein the effective amount ratio of component (B) to component (A) is 0.005 to 50.

9. The agent according to claim 1 or 2, which is an oral flora improver, a caries prevention / improvement agent, and a periodontal disease prevention / improvement agent.

10. Component (A): A nitrate ion supply compound, and Component (B): At least one selected from the group consisting of phytosterol, cholesterol, lanosterol, cholesteryl butyrate, β-sitosterol, γ-oryzanol, and dihydrocholesterol An oral composition containing the same.

11. The oral composition according to claim 10, which is a tablet, a drink, or an oral film.

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