Oral microbiota improver

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 technology]

[0002] The oral cavity is home to a wide variety of bacteria, and oral flora, consisting of colonies of these bacteria, exists in various locations, such as the tooth surfaces, gums, gingival margin, palate, buccal mucosa, and tongue surface and dorsum. This oral flora contains not only pathogenic bacteria involved in oral diseases such as dental caries and periodontal disease, but also non-pathogenic bacteria as resident bacteria. Maintaining a balance between these non-pathogenic and pathogenic bacteria is believed to be highly useful in preventing or suppressing the onset and progression of oral diseases.

[0003] In this context, as a measure to prevent and treat periodontal disease, which is one of the oral diseases, Patent Document 1 discloses a composition containing copper chlorophyllin salt as an active ingredient for eliminating periodontal disease bacteria present within oral cells, etc., in an attempt to purify the interior of the cells by killing periodontal disease bacteria that have invaded oral cells. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Japanese Patent Application Publication No. 2023-34874 Summary of the Invention [Problem to be solved by the invention]

[0005] However, the compositions described in the above patent documents are solely intended to kill periodontal disease bacteria, and no consideration has been given to selectively and effectively promoting the growth of Neisseria bacteria in the oral cavity.

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

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

[0008] In other words, the present invention provides an oral flora improving agent, a growth promoter for oral Neisseria and / or Neisseria spp. bacteria, and an agent for accelerating the oral flora improving effect, each of which contains one or more active ingredients selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives. [Effects of the Invention]

[0009] According to the present invention, the growth of Neisseria bacteria in the oral cavity can be selectively promoted and the growth of pathogenic bacteria can be selectively inhibited, thereby effectively improving the oral flora, and the improvement of the oral flora can be rapidly achieved.Furthermore, the present invention has the potential to selectively promote the growth of not only Neisseria bacteria but also Russia bacteria. DETAILED DESCRIPTION OF THE INVENTION

[0010] The present invention will be described in detail below. In the present invention, the term "oral flora" is also referred to as "oral (oral or oral cavity) flora" or "oral (oral or oral cavity) microbiota," and refers to a community formed by a wide variety of bacteria that inhabit the oral cavity. In the present invention, "improving" the oral flora means changing the bacterial composition of the oral flora to increase the proportion of non-pathogenic bacteria or decrease the proportion of pathogenic bacteria, thereby converting the state of the oral flora to a state that leads to a healthy oral environment. Specifically, it means selectively promoting the growth of Neisseria or, more specifically, Russia in the oral cavity, thereby increasing the proportion of Neisseria and Russia bacteria (hereinafter collectively referred to as "Neisseria, etc."). It also means selectively suppressing the growth of pathogenic bacteria, thereby increasing the proportion of Neisseria, etc. in the oral flora.

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

[0012] The oral flora-improving agent of the present invention contains, as an active ingredient, one or more compounds selected from chlorophyll metal complex derivatives and chlorophyll derivatives. Specific examples of chlorophyll metal complex derivatives include copper chlorophyllin sodium and iron chlorophyllin sodium, while examples of chlorophyll derivatives include chlorophyll a and chlorophyll b. Among these, chlorophyll complex derivatives, especially copper chlorophyllin sodium, are preferred from the viewpoint of enhancing the growth-promoting effect on Neisseria bacteria and other bacteria in the oral flora. Although the detailed mechanism of the oral flora improving effect exerted by the oral flora improving agent of the present invention is not clear, it has been found that such chlorophyll metal complex derivatives and chlorophyll derivatives can unexpectedly promote the selective growth of bacteria of the genus Neisseria and the like, rapidly increasing the proportion of such bacteria or reducing the proportion of pathogenic bacteria, thereby changing the bacterial composition of the oral flora, and furthermore, it has been found that they can rapidly exert the oral flora improving effect, leading to the discovery of an agent containing these as active ingredients. If the proportion of Neisseria bacteria and the like is increased by the oral flora-improving agent of the present invention, it will be possible to lead the oral environment to a healthy state.

[0013] The specific form of the oral flora-improving agent of the present invention is not particularly limited, as long as it contains one or more active ingredients selected from chlorophyll metal complex derivatives and chlorophyll derivatives, can be applied to the oral cavity, and can remain in the oral cavity for a predetermined period of time. Examples of such forms include non-edible preparations such as liquid dentifrices, toothpastes, lubricating dentifrices, powdered dentifrices, mouthwashes, mouth rinses, gargles, and mouth fresheners (such as mouth sprays); and edible preparations for the oral cavity such as lozenges, edible films, chewing gums, candies, gummy candies, tablets, granules, fine granules, powders, and capsules. Among these, mouthwashes, mouthwashes, toothpastes, powdered toothpastes, and water toothpastes are preferred from the viewpoint of efficiently and simply exerting the effect of improving the oral flora.

[0014] The amount of one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives to be administered to the oral cavity in the oral flora improving agent of the present invention is not particularly limited, and can be appropriately selected depending on various conditions, such as the total amount of the oral flora improving agent to be administered per day, the body type and symptoms of the subject, the form of the agent, etc. For example, the total amount of one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives to be administered per day to an adult may be 0.005 mg or more, 0.01 mg or more, or even 0.1 mg or more, and may be 10 mg or less, 8 mg or less, or even 5 mg or less.

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

[0016] In order to more effectively promote the growth of Neisseria bacteria and other bacteria in the oral flora, the oral flora-improving agent of the present invention preferably further contains, as active ingredients, one or more selected from chlorophyll metal complex derivatives and chlorophyll derivatives, and a nitrate ion-donating compound. The nitrate ion-donating compound is a compound that can release nitrate (NO3) ions when it comes into contact with water.

[0017] Although the detailed mechanism achieved by using one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives and a nitrate ion supplying compound as active ingredients in the oral flora improving agent of the present invention is not clear, it is believed that in the oral cavity, Neisseria bacteria and the like use the nitrate ion supplying compound to produce nitric oxide, and 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 flora and effectively improving the oral flora.

[0018] Specific examples of the nitrate ion supplying compound include one or more compounds selected from alkali metal nitrates and alkaline earth metal nitrates. More specifically, such nitrate ion supplying compounds include one or more selected from sodium nitrate, potassium nitrate, and calcium nitrate, with potassium nitrate being preferred.

[0019] When the oral flora-improving agent of the present invention contains one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives and a nitrate ion-donating compound as active ingredients, the proportion of these active ingredients can be appropriately selected depending on various conditions, such as the total daily dosage of the oral flora-improving agent, the body type and symptoms of the subject, and the formulation of the agent. For example, per 1 part by mass of one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives, the proportion of the nitrate ion-donating compound may be 3 parts by mass or more, 6 parts by mass or more, or even 8 parts by mass or more, or may be 1500 parts by mass or less, 900 parts by mass or less, or even 400 parts by mass or less. When the oral flora-improving agent of the present invention contains one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives and a nitrate ion-donating compound as active ingredients, the proportion of these active ingredients may be 3 to 1500 parts by mass, 6 to 900 parts by mass, or even 8 to 400 parts by mass.

[0020] Furthermore, the amount of the nitrate ion supplying compound to be applied to the oral cavity may be, for example, 1 mg or more, 5 mg or more, or even 10 mg or more, or 500 mg or less, or 300 mg or less, or even 200 mg or less, 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 according to the total daily application amount of the oral flora improving agent, the application amount of the chlorophyll metal complex salt derivative, the application amount of the chlorophyll derivative, etc.

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

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

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

[0025] From the viewpoint of more effectively promoting the growth of Neisseria bacteria and the like in the oral flora, 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 flora improving agent of the present invention is not particularly limited and may be appropriately adjusted according to the total daily dosage of the oral flora improving agent, the dosage of one or more compounds selected from chlorophyll metal complex derivatives and chlorophyll derivatives, or the dosage of the nitrate ion-supplying compound, etc. For example, the total content of these 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, based on the total amount of the oral flora improving agent of the present invention. The total content of these compounds 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 an organic acid salt is contained as the compound, the content thereof 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, and preferably 18% by mass or less, more preferably 8% by mass or less, and even more preferably 5% by mass or less, based on the total amount of the oral flora-improving agent of the present invention, from the viewpoint of effectively increasing the growth-promoting effect of Neisseria bacteria and the like in oral flora. 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, based on the total amount of the oral flora-improving agent of the present invention.

[0028] Furthermore, when glycyrrhetinic acid is contained as the compound, 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, and preferably 1% by mass or less, more preferably 0.5% by mass or less, and even more preferably 0.2% by mass or less, based on the total amount of the oral flora-improving agent of the present invention, from the viewpoint of effectively increasing the growth-promoting effect of Neisseria bacteria and the like in oral flora. The content of glycyrrhetinic acid 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, based on the total amount of the oral flora-improving agent of the present invention.

[0029] Furthermore, when a fluorine-containing compound is contained as the compound, its content can be appropriately adjusted depending on the total daily application amount of the oral flora-improving agent, and is, for example, preferably 850 ppm or more, more preferably 900 ppm or more, even more preferably 950 ppm or more, and preferably 5000 ppm or less, more preferably 2000 ppm or less, and even more preferably 1400 ppm or less, in terms of fluorine atoms. The content of the fluorine-containing compound 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, based on the total amount of the oral flora-improving agent of the present invention.

[0030] When the fluorine-containing compound is a fluoride ion-donating compound, the content of the fluoride ion-donating compound can be appropriately adjusted depending on the total daily application amount of the oral flora-improving agent, etc., but from the viewpoint of effectively achieving the oral flora-improving effect by promoting the growth of Neisseria bacteria, etc., it is preferably 0.1 to 1.5 mass%, more preferably 0.5 to 1.4 mass%, and even more preferably 0.7 to 1.3 mass% of the total amount of the oral flora-improving agent of the present invention. Furthermore, when the fluorine-containing compound is a monofluorophosphate ion-donating compound, the content of the monofluorophosphate ion-donating compound is preferably 0.2 to 4 mass%, more preferably 0.4 to 2 mass%, and even more preferably 0.6 to 1.5 mass% of the total amount of the oral flora-improving agent of the present invention.

[0031] In the oral flora improving agent of the present invention, when the active ingredients are one or more selected from chlorophyll metal complex derivatives and chlorophyll derivatives, and a nitrate ion supplying compound, and the compound contains a fluorine-containing compound, the mass ratio (NO3 / F) of the nitric acid (NO3) equivalent amount of the nitrate ion supplying compound to the fluorine atom equivalent amount of the fluorine-containing compound is, from the viewpoint of increasing the growth-promoting effect of Neisseria and other bacteria in the oral flora, preferably 0.5 or more, more preferably 2 or more, even more preferably 4 or more, still more preferably 8 or more, still more preferably 12 or more, still more preferably 14 or more, preferably 50 or less, more preferably 48 or less, still more preferably 46 or less, still more preferably 44 or less, still more preferably 42 or less, and still more preferably 38 or less. In the oral flora improving agent of the present invention, when one or more compounds selected from chlorophyll metal complex derivatives and chlorophyll derivatives and a nitrate ion supplying compound are used as active ingredients and the compound contains a fluorine-containing compound, the mass ratio (NO3 / F) of the nitric acid (NO3) equivalent amount of the nitrate ion supplying compound to the fluorine atom equivalent amount of the fluorine-containing compound is preferably 0.5 to 50, more preferably 2 to 48, even more preferably 4 to 46, still more preferably 8 to 44, still more preferably 12 to 42, and even more preferably 14 to 38.

[0032] In order to ensure a high oral flora-improving effect, it is preferable to limit the content of sulfate ester salts in the oral flora-improving agent of the present invention. 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 flora improving effect, the content of sulfate ester salts in the total amount of the oral flora improving agent of the present invention is preferably 0.1 mass% or less, more preferably 0.05 mass% or less, and even more preferably 0.01 mass% or less. Alternatively, it is preferable that the oral flora improving agent of the present invention does not contain sulfate ester salts.

[0034] It is preferable that the oral flora-improving agent of the present invention contains a limited amount of bactericide, from the viewpoint of avoiding a decrease in the effect of promoting the growth of Neisseria bacteria and ensuring a high oral flora-improving effect.

[0035] From the viewpoint of effectively enhancing the oral flora improving effect, the content of the bactericide in the total amount of the oral flora 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 flora improving agent of the present invention does not contain a bactericide.

[0036] Furthermore, among the disinfectants, it is preferable to limit the content of cationic disinfectants. Specific examples of cationic disinfectants include one or more selected from quaternary ammonium compounds and biguanide compounds. Examples of 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. Examples of 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 flora improving effect, the content of the cationic bactericide in the total amount of the oral flora 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. Alternatively, it is preferable that the oral flora improving agent of the present invention does not contain a cationic bactericide.

[0038] In addition to the above-mentioned ingredients, the oral flora improving agent of the present invention may contain solvents such as water and ethanol; pH adjusters; binders; humectants such as glycerin, propylene glycol, and polyethylene glycol; abrasives; sweeteners; foaming agents and foaming aids; preservatives; colorants; fragrances, etc., as appropriate, within the scope that does not inhibit the effects of the present invention, depending on various conditions such as the total daily dosage of the oral flora improving agent, the body type and symptoms of the subject, or the form of the agent.

[0039] In this way, by using one or more active ingredients selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives, the growth of Neisseria and other bacteria can be promoted, and the proportion of Neisseria and other bacteria in the oral flora can be effectively increased. Therefore, the active ingredients can be used to promote the growth of oral Neisseria and other bacteria, or as agents for accelerating the improvement of oral flora. Similarly, if one or more selected from chlorophyll metal complex derivatives and chlorophyll derivatives and a nitrate ion supply compound are used as active ingredients, the growth of Neisseria and other bacteria can be promoted, and the proportion of Neisseria and other bacteria in the oral flora can be effectively increased.Therefore, the composition can be used to promote the growth of oral Neisseria and other bacteria, or to accelerate the improvement of the oral flora.

[0040] In relation to the above-mentioned embodiment, the present invention further discloses the following oral flora improving agent. [1] An oral flora improving agent containing one or more active ingredients selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives. [2] The oral flora improving agent according to the above [1], wherein the chlorophyll metal complex salt derivative is preferably copper chlorophyllin sodium and / or iron chlorophyllin sodium. [3] The oral flora improving agent according to [1] or [2] above, wherein the chlorophyll derivative is chlorophyll a and / or chlorophyll b. [4] The oral flora improving agent of any one of [1] to [3] above, wherein the amount of one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives to be applied to the oral cavity may be 0.005 mg or more, or may be 0.01 mg or more, or may be 0.1 mg or more, or may be 10 mg or less, or may be 8 mg or less, or may be 5 mg or less, as the total amount of one or more selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives per adult per day.

[0041] [5] The oral flora improving agent according to any one of the above [1] to [4], further comprising a nitrate ion supplying compound as an active ingredient. [6] The oral flora improving agent of [5] above, wherein the nitrate ion supplying 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 flora improving agent of [5] or [6] above, wherein the amount of the nitrate ion supply compound is 3 parts by mass or more, or 6 parts by mass or more, or even 8 parts by mass or more, per 1 part by mass of chlorophyll metal complex salt derivatives and one or more selected from chlorophyll derivatives, and 1,500 parts by mass or less, or 900 parts by mass or less, or even 400 parts by mass or less. [8] The oral flora improving agent according to any one of [5] to [7] above, wherein the amount of the nitrate ion supplying compound to be applied to the oral cavity may be 1 mg or more, or may be 5 mg or more, or may be 10 mg or more, or may be 500 mg or less, or may be 300 mg or less, or may be 200 mg or less, as the nitrate ion supplying compound per adult per day.

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

[10] The oral flora-improving agent of 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, and preferably 5% by mass or less, more preferably 4% by mass or less, even more preferably 3% by mass or less, and still more preferably 2% by mass or less.

[11] The oral flora 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, and preferably 18% by mass or less, more preferably 8% by mass or less, even more preferably 5% by mass or less.

[12] The oral flora-improving agent according to any one of the above [9] to

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

[13] The oral flora improving agent according to any one of the above [9] to

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

[0043]

[14] The oral flora improving agent according to any one of the above [1] to

[13] , wherein the content of sulfate ester salt 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 the oral flora improving agent of the present invention preferably does not contain sulfate ester salt.

[15] The oral flora improving agent according to any one of [1] to

[14] above, which is a growth promoter for oral Neisseria and / or Neisseria spp.

[16] The oral flora improving agent according to any one of the above [1] to

[14] , which is an agent for accelerating the oral flora improving effect. [Example]

[0044] The present invention will be described in detail below with reference to the following examples. Unless otherwise specified in the tables, the content of each component is expressed in mass %.

[0045] <Test Example 1> According to the methods (1) to (4) below, the effect of improving the oral flora was evaluated using the growth-promoting effect of Neisseria bacteria in the dental plaque biofilm flora (increase ratio (fold) and increase amount (%)) as indicators.

[0046] (1) Plaque collection and pretreatment Dental plaque from healthy female teeth was collected using a dental scaler, placed in a 1.5 mL tube, and washed twice with PBS(-) by centrifugation at 7,000 g for 2 minutes at room temperature. The pellet containing plaque bacteria was suspended in 300 μL of CELLBANKER (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 copy number of the total bacterial rRNA16S gene in the plaque suspension using the DNA extraction method and real-time PCR described below.

[0047] (2) Preparation of a dental plaque biofilm model using dental plaque suspension culture Oral compositions were prepared using copper chlorophyllin sodium (manufactured by Tama Biochemical Co., Ltd.), chlorophyll (manufactured by Tokyo Chemical Industry Co., Ltd.), potassium nitrate (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), sodium citrate (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), β-glycyrrhetinic acid (manufactured by Alps Pharmaceutical Industries, Ltd.), sodium fluoride (manufactured by Stella Chemifa Co., Ltd.), sodium monofluorophosphate (manufactured by BK Giulini GmbH), with the remainder being water, according to the formula shown in Tables 3 to 5. Each of these was diluted 10-fold with Brain Heart Infusion (BHI) medium (Japan BD) to prepare evaluation samples. The evaluation samples were prepared by measuring the total bacterial rRNA16S gene copy number of 10 in dental plaque suspensions. 7 A culture medium containing dental plaque bacteria was prepared by diluting the culture medium to 100 copies / mL. 1 mL of the culture medium was added to a Poly-L-Lysine Coated 24-Well Microplate (AGC Techno Glass) and cultured at 37°C under microaerobic conditions using an Anaeropack Bikouki (Mitsubishi Gas Chemical Company, Inc.) for 48 hours to produce dental plaque biofilms.

[0048] (3) Extraction of genomic DNA from dental plaque biofilm The dental plaque biofilm attached to the bottom of the plate was thoroughly mixed with the culture supernatant by pipetting and then collected in a 1.5 mL tube. The plate was then centrifuged (14,000 rpm, room temperature for 10 minutes). The culture supernatant was removed, and the resulting precipitate was added to 200 μL / well of 20 mg / mL lysozyme (Fujifilm Wako Pure Chemical Industries, Ltd.) solution (containing 20 mM Tris-HCl (pH 8.0), 2 mM EDTA, and 1.2% Triton X-100). The plate was then 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 bacterial species was calculated by real-time PCR using Taqman probes. The primers and Taqman probes used and the reaction conditions for real-time PCR are shown in Tables 1 and 2.

[0049] [Table 1]

[0050] [Table 2]

[0051] (4) Calculation of the abundance ratio of each bacteria The abundance ratio (%) of each bacterium was calculated by first creating a calibration curve using PCR products with known copy numbers as standards, then calculating the copy number of the rRNA16S gene of each bacterium using the following formula. Percentage of each bacterium present (%) = (copy number of each bacterium) / (total bacterial copy number) × 100

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

[0053] Specifically, as shown in Table 3, Comparative Example 1 was selected as a control example for Examples 1 to 8. Furthermore, as shown in Table 4, Example 2 was selected as a control example for Example 9, Example 3 was selected as a control example for Example 10, Example 4 was selected as a control example for Example 11, Example 5 was selected as a control example for Example 12, Example 6 was selected as a control example for Examples 13 and 14, Example 7 was selected as a control example for Examples 15 and 16, and Example 8 was selected 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 carried out according to the following method. The results are shown in Tables 3 to 5.

[0054] Evaluation 1: Neisseria growth promotion effect (increase ratio (times)) Based on the quantitative bacterial counts obtained for each agent, the growth promoting effect of Neisseria bacteria (increase factor (fold)) was calculated using the following formula. Neisseria growth promotion effect (increase ratio (fold)) = (proportion of Neisseria bacteria present) / (the proportion of Neisseria present in the comparative example selected as the control)

[0055] [Table 3]

[0056] [Table 4]

[0057] [Table 5]

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

[0059] [Table 6]

[0060] <Test Example 3 [Example 60]> Mouthwashes were prepared according to the formulation shown in Table 7, with the remainder being purified water to make up 100% by mass, and their activity against bacterial growth in saliva was 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 bacteria One subject used the mouthwash three times a day (gargling 20 mL of mouthwash for 30 seconds) for three days, and the bacterial ratio in stimulated saliva was compared before and after use. Furthermore, from three days before using the mouthwash until the end of the test period, the subject was required to use the designated commercially available toothpaste and toothbrush to eliminate the influence of oral hygiene practices other than the use of the mouthwash. The stimulated saliva was collected by chewing commercially available paraffin wax gum for 3 minutes, and the leaked saliva was spat out into a plastic container at regular intervals. After collection, the saliva was promptly stored in a freezer at -20°C.

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

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

[0064] [Table 7]

Claims

1. An agent for improving oral flora, comprising one or more active ingredients selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives.

2. The oral flora improving agent according to claim 1, further comprising a nitrate ion supplying compound as an active ingredient.

3. 3. The oral flora-improving agent according to claim 1, further comprising one or more compounds selected from the group consisting of organic acid salts, glycyrrhetinic acid, and fluorine-containing compounds as active ingredients.

4. A growth promoter for oral Neisseria and / or Neisseria spp. bacteria, comprising one or more active ingredients selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives.

5. The growth promoter for oral Neisseria and / or Russia bacteria according to claim 4, further comprising a nitrate ion supplying compound as an active ingredient.

6. 6. The oral growth promoter for Neisseria and / or Neisseria spp. according to claim 4 or 5, further comprising one or more compounds selected from organic acid salts, glycyrrhetinic acid, and fluorine-containing compounds as active ingredients.

7. The growth promoter for oral Neisseria and / or Neisseria spp. bacteria according to claim 4 or 5, which has an inhibitory effect on the growth of oral pathogenic bacteria.

8. An agent for accelerating the action of improving oral flora, which comprises one or more active ingredients selected from chlorophyll metal complex salt derivatives and chlorophyll derivatives.

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