Oral microbiome improving agent

Abstract

To provide an oral microbiota improver that selectively and efficiently promotes growth of Neisseria and / or Rothia species in the oral microbiota, enabling effective improvement of the oral microbiota.SOLUTION: An oral microbiota improver comprising, as an active ingredient, one or two selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to an oral flora improving agent.

Background Art

[0002] A wide variety of bacteria inhabit the oral cavity in large numbers, and oral flora formed by these bacteria forming communities are present at various sites such as tooth surfaces, gums, gingival margins, palatal and buccal mucosa, tongue surfaces and dorsal tongue surfaces. In such oral flora, not only pathogenic bacteria involved in oral diseases such as dental caries and periodontal disease, but also non-pathogenic bacteria coexist as resident bacteria. Balancing these non-pathogenic bacteria and pathogenic bacteria is said to be highly useful in the onset, prevention, or inhibition of progression of oral diseases.

[0003] Under such circumstances, as a measure for the onset and prevention of periodontal disease, which is one of the oral diseases, Patent Document 1 discloses a composition containing a copper chlorophyllin salt as an active ingredient for eliminating periodontal disease bacteria present in oral cells and the like, and attempts have been made to purify the inside of cells by sterilizing periodontal disease bacteria that have invaded oral cells.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Summary of Invention

Problem to be Solved by the Invention

[0005] However, the composition described in the above patent document is solely intended to sterilize periodontal disease bacteria, and no investigations have been made at all regarding selectively and effectively promoting the growth of Neisseria bacteria in the oral cavity.

[0006] Therefore, the present invention relates to an oral microbiota improving agent that can selectively and effectively promote the growth of Neisseria species and / or Russula species in the oral microbiota, thereby effectively improving the oral microbiota. [Means for solving the problem]

[0007] Therefore, the inventors conducted diligent research to solve the above problems and, as a result, unexpectedly discovered an agent that can selectively and effectively promote the growth of Neisseria and Russula bacteria in the oral microbiota by using one or more selected chlorophyll metal complex salt derivatives and chlorophyll derivatives as active ingredients.

[0008] In other words, the present invention provides an oral microbiota improving agent, an oral Neisseria and / or Russula growth promoter, and an oral microbiota improving agent, comprising one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives as active ingredients. [Effects of the Invention]

[0009] According to the present invention, it is possible to selectively promote the growth of Neisseria species in the oral cavity and selectively suppress the growth of pathogenic bacteria, thereby effectively improving the oral microbiota, and furthermore, to rapidly exhibit this improvement in the oral microbiota. Moreover, it has the potential to selectively promote the growth of not only Neisseria species but also Russula species. [Modes for carrying out the invention]

[0010] The present invention will be described in detail below. In this invention, "oral microbiota" is also referred to as "oral (mouth or oral) flora" or "oral (mouth or oral) microbiota," and means a community formed by a wide variety of bacteria that inhabit the oral cavity. Furthermore, in this invention, "improving" the oral microbiota means changing the bacterial composition of the oral microbiota to a state that brings about a healthy oral environment by increasing the proportion of non-pathogenic bacteria or decreasing the proportion of pathogenic bacteria. Specifically, this means increasing the proportion of Neisseria species and Russian species (hereinafter collectively referred to as "Neisseria species, etc.") in the oral microbiota by selectively promoting the growth of Neisseria species or, more specifically, Russian species in the oral cavity. It also means increasing the proportion of Neisseria species, etc. in the oral microbiota by selectively suppressing the growth of pathogenic bacteria.

[0011] Here, Neisseria species are non-pathogenic oral commensal bacteria belonging to the genus Neisseria in the family Neisseriaceae. Specifically, these Neisseria species include Neisseria mucosa, Neisseria sicca, Neisseria flava, Neisseria subflava, Neisseria flavescens, and Neisseria elongata. Furthermore, the genus Rothia consists of Gram-positive cocci or bacilli belonging to the genus Rothia in the family Micrococcaceae, and are commensal bacteria of the oral cavity. The genus Rothia includes species such as Rothia aeria, Rothia mucilaginosa, and Rothia dentocariosa. In this invention, "pathogenic bacteria" specifically refers to bacteria of the genus Prevotella, Veillonella, Porphyromonas gingivalis (Pg), Fusobacterium nucleatum (Fn), or Tannerella forsythia.

[0012] The oral microbiota improving agent of the present invention comprises one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives as active ingredients. Specific examples of chlorophyll metal complex salt derivatives include copper chlorophyllin sodium and iron chlorophyllin sodium, while examples of chlorophyll derivatives include chlorophyll a and chlorophyll b. Among these, chlorophyll complex salt derivatives, particularly copper chlorophyllin sodium, are preferred from the viewpoint of increasing the growth-promoting effect of Neisseria species and other bacteria in the oral microbiota. Although the detailed mechanism of the oral microbiota improving effect achieved by the oral microbiota improving agent of the present invention is not yet clear, it has been found that such chlorophyll metal complex salt derivatives and chlorophyll derivatives can unexpectedly promote the selective growth of Neisseria species and other bacteria, rapidly increasing their proportion or decreasing the proportion of pathogenic bacteria, thereby altering the bacterial composition of the oral microbiota. Furthermore, it has been found that this can rapidly manifest its effect of improving the oral microbiota, leading to the discovery of an agent containing these as active ingredients. If the proportion of Neisseria species and other bacteria increases due to the oral microbiota improving agent of the present invention, it will be possible to guide the oral environment to a healthy state.

[0013] The specific form of the oral microbiota improving agent of the present invention is not particularly limited, as long as it contains one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives as an active ingredient, is applicable to the oral cavity, and can remain in the oral cavity for a predetermined period of time. Examples of such forms include non-edible formulations such as liquid toothpaste, paste toothpaste, moistened toothpaste, powder toothpaste, mouthwash, mouth rinse, gargle, and oral fresheners (mouth spray, etc.); and edible oral formulations such as lozenges, edible films, chewing gum, candy, gummy candy, tablets, granules, fine granules, powder, and capsules. In particular, mouthwashes, toothpastes, toothpaste powders, and toothpaste liquids are preferred from the viewpoint of efficiently and easily exhibiting the effect of improving the oral microbiome.

[0014] The amount of one or more chlorophyll metal complex salt derivatives and chlorophyll derivatives selected from the oral microbiota improving agent of the present invention applied to the oral cavity is not particularly limited and can be appropriately selected according to various conditions such as the total amount of oral microbiota improving agent applied per day, the body type and symptoms of the target person, or the form of the agent. For example, the total amount of one or more chlorophyll metal complex salt derivatives and chlorophyll derivatives selected from one or more chlorophyll derivatives per day for one adult may be 0.005 mg or more, 0.01 mg or more, 0.1 mg or more, 10 mg or less, 8 mg or less, or 5 mg or less.

[0015] The total content of one or more chlorophyll metal complex salt derivatives and chlorophyll derivatives selected from the oral microbiota improving agent of the present invention is not particularly limited and may be appropriately adjusted according to the total daily application amount of the oral microbiota improving agent.

[0016] From the viewpoint of more effectively promoting the growth of Neisseria species and other bacteria in the oral microbiota of the present invention, it is preferable that the oral microbiota improving agent further contains one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives, and a nitrate ion supply compound as active ingredients. The nitrate ion supply compound is a compound that can release nitrate (NO3) ions when it comes into contact with water.

[0017] In the oral microbiota improving agent of the present invention, although the detailed mechanism by which one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives, and a nitrate ion supply compound are used as active ingredients is not yet clear, it is believed that Neisseria bacteria and the like utilize the nitrate ion supply compound to produce nitric oxide in the oral cavity, and that the synergistic effect with one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives greatly contributes to promoting the growth of Neisseria bacteria and the like, thereby rapidly increasing the proportion of Neisseria bacteria and the like in the oral microbiota and effectively improving the oral microbiota.

[0018] Examples of nitrate ion-supplying compounds include one or more selected from alkali metal nitrates and alkaline earth metal nitrates. More specifically, such nitrate ion supply compounds include one or more selected from sodium nitrate, potassium nitrate, and calcium nitrate. Among these, potassium nitrate is preferred.

[0019] In the oral flora improving agent of the present invention, when one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives and a nitrate ion supplying compound are used as active ingredients, the ratio of these active ingredients can be appropriately selected according to various conditions such as the total application amount of the oral flora improving agent per day, the body type and symptoms of the subject to be applied, or the form of the agent. For example, relative to 1 part by mass of one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives, the nitrate ion supplying compound may be 3 parts by mass or more, may be 6 parts by mass or more, may be 8 parts by mass or more, may be 1500 parts by mass or less, may be 900 parts by mass or less, and may be 400 parts by mass or less. In the oral flora improving agent of the present invention, when one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives and a nitrate ion supplying compound are used as active ingredients, the ratio of these active ingredients may be 3 to 1500 parts by mass, may be 6 to 900 parts by mass, and may be 8 to 400 parts by mass.

[0020] In addition, the application amount of the nitrate ion supplying compound into the oral cavity may be, for example, 1 mg or more, may be 5 mg or more, may be 10 mg or more, may be 500 mg or less, may be 300 mg or less, and may be 200 mg or less, as the nitrate ion supplying compound per adult per day.

[0021] The content of the nitrate ion supplying compound in the oral flora improving agent of the present invention is not particularly limited, and may be appropriately adjusted in accordance with the total application amount of the oral flora improving agent per day, or the application amount of the chlorophyll metal complex salt derivative, the application amount of the chlorophyll derivative, and the like.

[0022] From the viewpoint of enhancing the growth promoting effect on bacteria of the genus Neisseria and the like in the oral flora, the oral flora improving agent of the present invention preferably further comprises one or more compounds selected from organic acid salts, glycyrrhetinic acid, and fluorine-containing compounds as active ingredients. Specific examples of the organic acid salt include one or two or more selected from citrate, gluconate, succinate, fumarate, lactate, malate, and tartrate. Among these, citrate, gluconate, succinate, fumarate, lactate, and malate are preferred, and citrate is more preferred. Examples of the salt include metal salts such as sodium, potassium, magnesium, aluminum, and calcium. Among these, sodium salt and potassium salt are preferred. More specifically, among these organic acid salts, sodium citrate is preferred from the viewpoint of effectively improving the oral microbiota.

[0023] Specific examples of glycyrrhetinic acid include α-glycyrrhetinic acid, β-glycyrrhetinic acid and the like. Among these, β-glycyrrhetinic acid is preferred.

[0024] Specific examples of the fluorine-containing compound include one or two or more selected from a fluoride ion supplying compound and a monofluorophosphate ion supplying compound. The fluoride ion supplying compound is a fluorine-containing compound other than compounds that supply monofluorophosphate ions. Examples of such a fluoride ion supplying compound include one or two or more selected from sodium fluoride, stannous fluoride, potassium fluoride, zinc fluoride, betaine fluoride, stannous fluoride alanine, sodium fluorosilicate, hexylamine fluoride and the like. Among these, sodium fluoride or stannous fluoride is preferred. Specific examples of the monofluorophosphate ion supplying compound include one or two or more selected from sodium monofluorophosphate, potassium monofluorophosphate, magnesium monofluorophosphate, calcium monofluorophosphate and the like. Among these, sodium monofluorophosphate is preferred. Among these, as the fluorine-containing compound, a monofluorophosphate ion supplying compound is preferred from the viewpoint of effectively improving the oral microbiota.

[0025] From the viewpoint of more effectively promoting the growth of Neisseria species and other bacteria in the oral microbiota, these compounds are preferably one or more selected from glycyrrhetinic acid and fluorine-containing compounds, and more preferably contain glycyrrhetinic acid.

[0026] The total content of these compounds in the oral microbiota improving agent of the present invention is not particularly limited and can be appropriately adjusted according to the total daily application amount of the oral microbiota improving agent, or the application amount of one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives, or the application amount of nitrate ion supply compounds, etc. For example, the total content of these compounds in the total amount of the oral microbiota improving agent of the present invention is preferably 0.005% by mass or more, preferably 0.02% by mass or more, preferably 0.3% by mass or more, more preferably 0.5% by mass or more, preferably 5% by mass or less, more preferably 4% by mass or less, even more preferably 3% by mass or less, and even more preferably 2% by mass or less. The total content of these compounds in the total amount of the oral microbiota improving agent of the present invention is preferably 0.005 to 5% by mass, more preferably 0.02 to 4% by mass, even more preferably 0.3 to 3% by mass, and even more preferably 0.5 to 2% by mass.

[0027] More specifically, when the above compound includes an organic acid salt, its content is preferably 0.1% by mass or more, more preferably 0.5% by mass or more, even more preferably 0.8% by mass or more, preferably 18% by mass or less, more preferably 8% by mass or less, and even more preferably 5% by mass or less, in the total amount of the oral microbiota improving agent of the present invention, from the viewpoint of effectively increasing the growth-promoting effect of Neisseria species and the like in the oral microbiota. Furthermore, the content of the organic acid salt is preferably 0.1 to 18% by mass, more preferably 0.5 to 8% by mass, and even more preferably 0.8 to 5% by mass, in the total amount of the oral microbiota improving agent of the present invention.

[0028] Furthermore, when the above compound includes glycyrrhetinic acid, its content is preferably 0.005% by mass or more, more preferably 0.01% by mass or more, even more preferably 0.02% by mass or more, preferably 1% by mass or less, more preferably 0.5% by mass or less, and even more preferably 0.2% by mass or less, in the total amount of the oral microbiota improving agent of the present invention, from the viewpoint of effectively increasing the growth-promoting effect of Neisseria species and the like in the oral microbiota. The glycyrrhetinic acid content is preferably 0.005 to 1% by mass, more preferably 0.01 to 0.5% by mass, and even more preferably 0.02 to 0.2% by mass, in the total amount of the oral microbiota improving agent of the present invention.

[0029] Furthermore, when the above compound includes a fluorine-containing compound, its content can be appropriately adjusted according to the total amount of the oral microbiota improving agent applied per day, for example, in terms of fluorine atoms, it is preferably 850 ppm or more, more preferably 900 ppm or more, even more preferably 950 ppm or more, preferably 5000 ppm or less, more preferably 2000 ppm or less, and even more preferably 1400 ppm or less. The content of the fluorine-containing compound in the total amount of the oral microbiota improving agent of the present invention is preferably 850 to 5000 ppm, more preferably 900 to 2000 ppm, and even more preferably 950 to 1400 ppm in terms of fluorine atoms.

[0030] Furthermore, when the fluorine-containing compound is a fluoride ion-supplying compound, the content of such fluoride ion-supplying compound can be appropriately adjusted according to the total amount of the oral microbiota improving agent applied per day, etc. However, from the viewpoint of effectively bringing about the oral microbiota improving effect through the growth-promoting action of Neisseria species and the like, the content of such compound in the total amount of the oral microbiota improving agent of the present invention is preferably 0.1 to 1.5% by mass, more preferably 0.5 to 1.4% by mass, and even more preferably 0.7 to 1.3% by mass. Furthermore, when the fluorine-containing compound is a monofluorophosphate ion-supplying compound, the content of such monofluorophosphate ion-supplying compound in the total amount of the oral microbiota improving agent of the present invention is preferably 0.2 to 4% by mass, more preferably 0.4 to 2% by mass, and even more preferably 0.6 to 1.5% by mass.

[0031] In the oral microbiota improving agent of the present invention, one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives, and a nitrate ion supplying compound are used as active ingredients, and the compound also contains a fluorine-containing compound, the mass ratio (NO3 / F) of the amount of nitrate (NO3) of the nitrate ion supplying compound to the amount of fluorine atoms of the fluorine-containing compound is preferably 0.5 or higher, more preferably 2 or higher, even more preferably 4 or higher, even more preferably 8 or higher, even more preferably 12 or higher, even more preferably 14 or higher, preferably 50 or lower, more preferably 48 or lower, even more preferably 46 or lower, even more preferably 44 or lower, even more preferably 42 or lower, and even more preferably 38 or lower. Furthermore, in the oral microbiota improving agent of the present invention, when one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives and a nitrate ion supply compound are used as active ingredients, and the compound includes a fluorine-containing compound, the mass ratio (NO3 / F) of the amount of nitrate (NO3) of the nitrate ion supply compound to the amount of fluorine atoms of the fluorine-containing compound is preferably 0.5 to 50, more preferably 2 to 48, even more preferably 4 to 46, even more preferably 8 to 44, even more preferably 12 to 42, and even more preferably 14 to 38.

[0032] Furthermore, in the oral microbiota improving agent of the present invention, it is preferable to limit the content of sulfate ester salts from the viewpoint of ensuring a high oral microbiota improving effect. Examples of such sulfate ester salts include one or more selected from alkyl sulfates, alkenyl sulfates, polyoxyalkylene alkyl ether sulfates, polyoxyalkylene alkenyl ether sulfates, and polyoxyalkylene alkylphenyl ether sulfates.

[0033] From the viewpoint of effectively enhancing the oral microbiota improving effect, the content of sulfate ester salts in the total amount of the oral microbiota improving agent of the present invention is preferably 0.1% by mass or less, more preferably 0.05% by mass or less, and even more preferably 0.01% by mass or less, or it is preferable that the oral microbiota improving agent of the present invention does not contain sulfate ester salts.

[0034] In order to avoid a decrease in the effect of promoting the growth of Neisseria species and other bacteria in the oral microbiota improving agent of the present invention and to ensure a high oral microbiota improving effect, it is preferable to limit the content of the bactericide.

[0035] From the viewpoint of effectively enhancing the oral microbiome improving effect, the amount of bactericide in the total amount of the oral microbiome improving agent of the present invention is preferably less than 0.5% by mass, more preferably 0.1% by mass or less, and even more preferably 0.05% by mass or less, or it is preferable that the oral microbiome improving agent of the present invention does not contain a bactericide.

[0036] Furthermore, it is preferable to limit the content of cationic disinfectants among the disinfectants. Cationic disinfectants specifically include, for example, one or more selected from quaternary ammonium compounds and biguanide compounds. Cationic disinfectants belonging to quaternary ammonium compounds include one or more selected from cetylpyridinium chloride, benzethonium chloride, depotassium chloride, benzalkonium chloride, alkyldimethylammonium chloride, alkyltrimethylammonium chloride, and methylbenzethonium chloride. Cationic disinfectants belonging to biguanide compounds include one or more selected from chlorhexidine and its salts, such as chlorhexidine gluconate and chlorhexidine hydrochloride.

[0037] From the viewpoint of effectively enhancing the oral microbiota improving effect, the content of cationic bactericide in the total amount of the oral microbiota improving agent of the present invention is preferably less than 0.5% by mass, more preferably 0.1% by mass or less, and even more preferably 0.05% by mass or less, or it is preferable that the oral microbiota improving agent of the present invention does not contain a cationic bactericide.

[0038] The oral microbiota improving agent of the present invention may contain, in addition to the above-mentioned components, solvents such as water or ethanol; pH adjusters; binders; wetting agents such as glycerin, propylene glycol, or polyethylene glycol; abrasives; sweeteners; foaming agents or foaming aids; preservatives; colorants; fragrances, etc., as appropriate, depending on various conditions such as the total daily application amount of the oral microbiota improving agent, the body type and symptoms of the target person, or the form of the agent, as long as it does not inhibit the effects of the present invention.

[0039] Thus, by using one or more chlorophyll metal complex salt derivatives and chlorophyll derivatives as active ingredients, it is possible to promote the growth of Neisseria species and effectively increase the proportion of Neisseria species in the oral microbiota. Therefore, it can be widely used as a growth promoter for Neisseria species in the oral cavity or as an agent to accelerate the improvement of the oral microbiota. Similarly, if one or more chlorophyll metal complex salt derivatives and chlorophyll derivatives selected from these, along with a nitrate ion supply compound, are used as active ingredients, it is possible to promote the growth of Neisseria species and effectively increase the proportion of Neisseria species in the oral microbiota. Therefore, it can be widely used as a growth promoter for Neisseria species in the oral cavity, or as an agent to accelerate the improvement of the oral microbiota.

[0040] With regard to the embodiments described above, the present invention further discloses the following oral microbiota improving agent. [1] An oral microbiota improving agent comprising one or more chlorophyll metal complex salt derivatives and chlorophyll derivatives selected from among the active ingredients. [2] The oral microbiota improving agent according to [1] above, wherein the chlorophyll metal complex salt derivative is either copper chlorophyllin sodium and / or iron chlorophyllin sodium. [3] The oral microbiota improving agent according to [1] or [2] above, wherein the chlorophyll derivative is chlorophyll a and / or chlorophyll b. [4] The amount of one or more chlorophyll metal complex salt derivatives and chlorophyll derivatives selected from the above-mentioned oral microbiota improving agent may be any one of the above [1] to [3], where the total amount of one or more chlorophyll metal complex salt derivatives and chlorophyll derivatives selected from the above-mentioned oral microbiota improving agent may be 0.005 mg or more per adult per day, or 0.01 mg or more, or 0.1 mg or more, or 10 mg or less, or 8 mg or less, or 5 mg or less.

[0041] [5] Furthermore, an oral microbiota improving agent according to any one of the above [1] to [4], wherein a nitrate ion supply compound is used as the active ingredient. [6] The oral microbiota improving agent according to [5] above, wherein the nitrate ion supply compound is preferably one or more selected from alkali metal nitrates and alkaline earth metal nitrates, more preferably one or more selected from sodium nitrate, potassium nitrate and calcium nitrate, and even more preferably potassium nitrate. [7] The oral microbiota improving agent according to [5] or [6] above, wherein the nitrate ion supplying compound may be 3 parts by mass or more, 6 parts by mass or more, 8 parts by mass or more, 1500 parts by mass or less, 900 parts by mass or less, or 400 parts by mass or less, with respect to 1 part by mass of a chlorophyll metal complex salt derivative and one or more selected from chlorophyll derivatives. [8] The amount of nitrate ion supplying compound applied to the oral cavity may be 1 mg or more per adult per day, 5 mg or more, 10 mg or more, 500 mg or less, 300 mg or less, or 200 mg or less, as an oral microbiota improving agent according to any one of the above [5] to [7].

[0042] [9] Furthermore, an oral microbiota improving agent according to any one of the above [1] to [8], comprising one or more compounds selected from organic acid salts, glycyrrhetinic acid, and fluorine-containing compounds as an active ingredient, preferably comprising one or more compounds selected from glycyrrhetinic acid and fluorine-containing compounds as an active ingredient, and more preferably comprising glycyrrhetinic acid as an active ingredient.

[10] An oral microbiota improving agent according to any one of the above [1] to [9], wherein the total content of one or more compounds selected from organic acid salts, glycyrrhetinic acid, and fluorine-containing compounds is preferably 0.005% by mass or more, preferably 0.02% by mass or more, preferably 0.3% by mass or more, more preferably 0.5% by mass or more, preferably 5% by mass or less, more preferably 4% by mass or less, even more preferably 3% by mass or less, and even more preferably 2% by mass or less.

[11] The oral microbiota improving agent according to [9] or

[10] above, wherein the content of the organic acid salt is preferably 0.1% by mass or more, more preferably 0.5% by mass or more, even more preferably 0.8% by mass or more, preferably 18% by mass or less, more preferably 8% by mass or less, and even more preferably 5% by mass or less.

[12] An oral microbiota improving agent according to any one of the above [9] to

[11] , wherein the glycyrrhetinic acid content is preferably 0.005% by mass or more, more preferably 0.01% by mass or more, even more preferably 0.02% by mass or more, preferably 1% by mass or less, more preferably 0.5% by mass or less, and even more preferably 0.2% by mass or less.

[13] An oral microbiota improving agent according to any one of the above [9] to

[12] , wherein the content of the fluorine-containing compound is preferably 850 ppm or more, more preferably 900 ppm or more, even more preferably 950 ppm or more, preferably 5000 ppm or less, more preferably 2000 ppm or less, and even more preferably 1400 ppm or less, in terms of fluorine atoms.

[0043]

[14] The oral microbiota improving agent of the present invention preferably contains 0.1% by mass or less, more preferably 0.05% by mass or less, and even more preferably 0.01% by mass or less, or preferably does not contain sulfate ester salts, according to any one of the above [1] to

[13] .

[15] An oral microbiota improving agent that is a growth promoter of Neisseria species and / or Russula species in the oral cavity, as described in any one of [1] to

[14] above.

[16] An oral microbiome improving agent that is an agent that accelerates the action of improving the oral microbiome, the oral microbiome improving agent of any one of the above [1] to

[14] . [Examples]

[0044] The present invention will be described in detail below based on examples. Unless otherwise specified in the table, the content of each component is expressed in mass percent.

[0045] <Test Example 1> The effect of improving the oral microbiota was evaluated using the method described in (1) to (4) below, with the growth-promoting effect of Neisseria species in dental plaque biofilm microbiota (increase ratio (times) and increase amount (%)) as indicators.

[0046] (1) Plaque collection and pretreatment Plaque was collected from the tooth surface of healthy women using a dental scaler, placed in a 1.5 mL tube, and washed twice with PBS(-) at 7,000 g for 2 minutes at room temperature. The pellet containing plaque bacteria was suspended in 300 μL of CELLBANKER (manufactured by Nippon Zenyaku Kogyo Co., Ltd.), and the prepared plaque suspension was stored at -80°C until use. A portion of this plaque suspension was used to measure the total bacterial rRNA16S gene copy number in the plaque suspension using the DNA extraction method and real-time PCR method described later.

[0047] (2) Preparation of a plaque biofilm model by culturing plaque suspension Oral compositions were prepared using the formulations shown in Tables 3-5, with copper chlorophyllin sodium (Tama Biochemical Co., Ltd.), chlorophyll (Tokyo Chemical Industries Co., Ltd.), potassium nitrate (Fujifilm Wako Pure Chemical Industries, Ltd.), sodium citrate (Fujifilm Wako Pure Chemical Industries, Ltd.), β-glycyrrhetinic acid (Alps Pharmaceutical Co., Ltd.), sodium fluoride (Stella Chemifa Co., Ltd.), and sodium monofluorophosphate (BK Giulini GmbH), with the remainder being water. Each of these compositions was diluted 10-fold in Brain Heart Infusion (BHI) medium (BD Japan) and used as evaluation samples. In the evaluation samples, the total bacterial rRNA16S gene copy number in the plaque suspension was 10 7 A culture medium containing plaque bacteria was prepared by diluting the solution to a copy / mL ratio. One mL of the prepared culture medium was added to each Poly-L-Lysine Coated 24-Well Microplate (AGC TECHNO GLASS), and the cells were incubated at 37°C under microaerophilic conditions using an Anaeropack Bikouki (manufactured by Mitsubishi Gas Chemical Co., Ltd.) for 48 hours to produce plaque biofilm.

[0048] (3) Extraction of genomic DNA from dental plaque biofilm The plaque biofilm adhering to the bottom of the plate was thoroughly mixed with the culture supernatant by pipetting and then collected in a 1.5 mL tube. Next, it was centrifuged (14000 rpm at room temperature for 10 minutes), and 20 mg / mL lysozyme solution (20 mM Tris-HCl (pH 8.0), 2 mM MEDTA, 1.2% Triton X-100) was added to the precipitate after removing the culture supernatant at a rate of 200 μL / well, and incubated at 37°C for 60 minutes with shaking (160 rpm). Genomic DNA was extracted using the Dneasy Blood & Tissue kit (QIAGEN), and the proportion of each bacterium was calculated by real-time PCR using a Taqman probe. Tables 1 and 2 show the primers and Taqman probes used, as well as the reaction conditions for real-time PCR.

[0049] [Table 1]

[0050] [Table 2]

[0051] (4) Calculation of the relative abundance of each bacterium The percentage of each bacterium was determined by first creating a calibration curve using PCR products with known copy numbers as standards, then calculating the copy number of the rRNA16S gene for each bacterium, and finally determining the percentage using the following formula. The percentage of each bacterium (%) = (copy number of each bacterium) / (total bacterial copy number) × 100

[0052] [Examples 1-39, Comparative Example 1] Oral compositions were prepared according to the formulations shown in Tables 3-5, with the remainder being purified water, making a total volume of 100% by mass. The obtained oral compositions were diluted 10-fold in BHI medium to produce evaluation samples, and the bacteria were quantified according to (1)-(4) above. For Examples 1-8, samples without the additive were selected as comparative examples. For Examples 9-39, examples containing only the same amount of copper chlorophyllin sodium were selected as control examples, depending on the amount of copper chlorophyllin sodium in each example, and these were used as the basis for evaluation in Evaluation 1 below.

[0053] Specifically, as shown in Table 3, Comparative Example 1 was selected as a control example for Examples 1 to 8. Also, as shown in Table 4, Example 2 was selected as a control example for Example 9, Example 3 as a control example for Example 10, Example 4 as a control example for Example 11, Example 5 as a control example for Example 12, Example 6 as a control example for Examples 13 and 14, Example 7 as a control example for Examples 15 and 16, and Example 8 as a control example for Example 17. Furthermore, as shown in Table 5, Example 3 was selected as a control example for Examples 18 to 27, including Example 10, Example 4 was selected as a control example for Examples 28 to 36, including Example 11, and Example 6 was selected as a control example for Examples 37 to 39, including Example 14. Next, Evaluation 1 was performed according to the following method. The results are shown in Tables 3-5.

[0054] 《Evaluation 1: Neisseria fungal growth promoting effect (increase ratio (times))》 Based on the quantitative values ​​of bacteria obtained for each agent, the value of the growth-promoting effect (increase ratio (times)) on Neisseria species was calculated using the following formula. Neisseria growth-promoting effect (increase ratio (times)) = (proportion of Neisseria bacteria present) (Percentage of Neisseria species present in comparative examples selected as control cases)

[0055] [Table 3]

[0056] [Table 4]

[0057] [Table 5]

[0058] <Test Example 2 [Examples 40-59, Comparative Examples 2-3]> For Examples 40-49 and Comparative Example 2, the culture time was 12 hours, and for Examples 50-59 and Comparative Example 3, the culture time was 24 hours. The effect of improving the microbial flora was evaluated in the same manner as in Test 1. For Examples 40-49, Comparative Example 2 was selected as a comparative example, and for Examples 50-59, Comparative Example 3 was selected as a comparative example and used as the basis for evaluation 1 above. The results are shown in Table 6.

[0059] [Table 6]

[0060] <Test Example 3 [Example 60]> A mouthwash was prepared according to the formulation shown in Table 7, with the remainder being purified water to make up 100% by mass. The bacterial growth activity in saliva was then evaluated according to the methods shown in (1) and (2) below. The results are shown in Table 7.

[0061] (1) Calculation of the ratio of each bacterium Each subject used a mouthwash three times a day (20 mL of mouthwash gargled for 30 seconds) for three days, and the bacterial ratio in saliva before and after use was compared. Furthermore, from three days before mouthwash use until the end of the study period, subjects used a designated commercially available toothpaste and toothbrush to eliminate the influence of oral hygiene practices other than mouthwash use. Stimulated saliva was collected by having participants spit out any saliva that leaked out while chewing commercially available paraffin wax gum for 3 minutes, and then immediately storing the collected saliva in a -20°C freezer.

[0062] Next, 500 μL of saliva was collected after storage and centrifuged (14,000 rpm at room temperature for 10 minutes). Genomic DNA was extracted from the precipitate after removing the culture supernatant using the same method as for extracting genomic DNA from dental plaque biofilm as described above, and the proportion of each bacterium was calculated using real-time PCR with a Taqman probe. In addition, the primers and Taqman probes shown in Table 1 were used, as in Test Example 1 above, and the real-time PCR reaction conditions shown in Table 2 were followed.

[0063] (2) Calculation of the relative growth rate of each bacterium The percentage of each bacterium was determined by first creating a calibration curve using PCR products with known copy numbers as standards, then calculating the copy number of the rRNA16S gene for each bacterium, and finally determining the percentage using the following formula. The percentage of each bacterium (%) = (copy number of each bacterium) / (total bacterial copy number) × 100 Furthermore, the relative growth rate of each bacterium was calculated using the following formula. Relative increase rate of each bacterium (%) = (Percentage of each bacterium present after 3 days of use) / (Percentage of each bacterium present before use) × 100 Furthermore, bacteria with a relative growth rate of 100% or more were considered to have a growth-promoting effect, while those with a relative growth rate of less than 100% were considered to have a growth-inhibiting effect.

[0064] [Table 7]

Claims

1. A growth promoter for Neisseria species and / or Russula species in the oral cavity, comprising copper chlorophyllin sodium as an active ingredient.

2. Furthermore, the oral growth promoter for Neisseria species and / or Russula species according to claim 1, comprising a nitrate ion supply compound as an active ingredient.

3. Furthermore, the oral growth promoter for Neisseria species and / or Russula species according to claim 1 or 2 comprises one or more compounds selected from organic acid salts, glycyrrhetinic acid, and fluorine-containing compounds as active ingredients.

4. An oral growth promoter of Neisseria species and / or Russula species according to claim 1 or 2, which has an inhibitory effect on the growth of oral pathogenic bacteria.

5. The oral growth promoter for Neisseria species and / or Russula species according to claim 1, wherein the content of the active ingredient is 0.005% by mass or more and 0.5% by mass or less.

6. The oral growth promoter for Neisseria species and / or Russula species according to Claim 2, wherein the nitrate ion supply compound is 3 to 1500 parts by mass per 1 part by mass of copper chlorophyllin sodium.

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