Oral Neisseria mucosa enhancer

Abstract

To provide a material that is useful for increasing the Neisseria mucosa content in the oral cavity.SOLUTION: An oral Neisseria-mucosa-increasing agent has, as active ingredients, a nitrate-ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an agent for increasing Neisseria mucosa in the oral cavity. [Background technology]

[0002] In the human body, microorganisms form resident flora based on complex interactions and inhabit various parts of the body. The composition of the resident flora varies between individuals, and in recent years, many studies have pointed out the relationship between these individual differences and the health status of the host, attracting attention. Like the intestine, the oral cavity is a microbial ecosystem in which numerous types of bacteria coexist. Several pathogenic bacteria have been identified as responsible for oral infectious diseases such as dental caries and periodontal disease, but these bacteria also coexist in the oral cavity as part of the normal ecosystem. Therefore, it is speculated that interactions with other resident bacteria play a major role in the development of disease. While the traditional goal in preventing and treating oral disease has been to eliminate pathogenic bacteria that directly cause disease, it is believed that inducing a resident flora that makes it difficult for pathogenic bacteria to thrive could also be an approach to preventing oral disease.

[0003] Neisseria mucosa is known to exist as a non-pathogenic normal bacterium in the oral cavity, along with other Neisseria species such as Neisseria sicca, Neisseria flava, Neisseria subflava, Neisseria flavescens, and Neisseria elongata. The present applicant has recently discovered that Neisseria mucosa cells or cultures thereof suppress the infection of gingival cells by periodontal pathogens and are therefore useful for the prevention or treatment of periodontal disease (Patent Document 1). In addition, Neisseria bacteria possess nitrate reductase and nitrite reductase, which convert nitrate ions (NO3 - ) to nitrite ions (NO2 -It has been reported that nitrate ions have the ability to reduce nitrate to nitrite and nitric oxide (NO) (Non-Patent Document 1). When nitrate ions are orally ingested, a portion of the nitrate ions absorbed into the body migrates from the blood into saliva at high concentrations, where they are reduced to nitrite by nitrate-reducing bacteria normally present in the oral cavity. Nitric oxide (NO) is then produced enzymatically or non-enzymatically in the stomach and various tissues. Nitric oxide is an important compound that exerts various functions throughout the body, including intercellular signaling, antibacterial activity, biological defense against pathogens, vasodilatory activity, and muscle contractility. It is particularly expected to have antibacterial activity against periodontal pathogens, blood pressure reduction due to improved vascular function, and motor function improvement due to improved muscle contractility (Non-Patent Documents 2-5). Therefore, it is useful to increase the oral content of Neisseria bacteria.

[0004] On the other hand, potassium ions have the effect of reducing the hypersensitivity of dental pulp nerves, and potassium nitrate and the like are known to be effective against tooth hypersensitivity (Non-Patent Document 6). In addition, many sugar alcohols have the effect of promoting saliva secretion and tooth remineralization, and it has been reported that erythritol, xylitol, and sorbitol have caries prevention effects (Non-Patent Document 7). It has also been reported that sugar alcohols such as erythritol have the effect of inhibiting biofilm formation (Patent Document 2). However, the effects of potassium nitrate and sugar alcohols on oral Neisseria bacteria are completely unknown. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Patent application 2020-200517 [Patent Document 2] Japanese Patent Application Laid-Open No. 2005-29484 [Non-patent literature]

[0006] [Non-Patent Document 1] Barth KR et al., Microbiology (2009),155:4093-4103 [Non-patent document 2] Carlstrom M et al., Acta Physiol (Oxf).(2018),224:e13080 [Non-patent document 3] Masaru Shibata, Agriculture and Science, April 2019, 1-8 [Non-patent document 4] Omar SA et al., J Intern Med.(2016),279:315-336 [Non-patent document 5] CJ Backlund et al., J Dent Res 93(11):1089-1094, 2014 [Non-patent document 6] Poulsen S et al.,Cochrane Database Syst Rev.;(2006):CD001476 [Non-Patent Document 7] Peter de Cock P et al.,Int J Dent. (2016) ,2016:9868421. Summary of the Invention [Problem to be solved by the invention]

[0007] The present invention relates to providing a material useful for increasing the content of Neisseria mucosa in the oral cavity. [Means for solving the problem]

[0008] The inventors have discovered that by combining a nitrate ion supplying compound with a specific sugar alcohol, the content of Neisseria mucosa and the content of Neisseria bacteria in the oral cavity can be increased, and this is useful for increasing nitric oxide and preventing or treating periodontal disease.

[0009] That is, the present invention relates to the following 1) to 8). 1) An oral Neisseria mucosa increasing agent having as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. 2) An agent for increasing the number of oral Neisseria bacteria, the agent comprising as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. 3) An oral flora improving agent containing as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. 4) A periodontal pathogen infection inhibitor comprising as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. 5) A preventive or ameliorating agent for periodontal disease, which comprises as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. 6) A nitric oxide enhancer comprising a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol as active ingredients. 7) A hypertension prevention or amelioration agent comprising as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. 8) A motor function improving or enhancing agent comprising as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. [Effects of the Invention]

[0010] The present invention can increase the content of Neisseria mucosa and Neisseria bacteria in the oral cavity, thereby enabling the prevention or treatment of periodontal disease. Furthermore, the supply of nitric oxide can improve vascular function and skeletal muscle contractility, thereby preventing or ameliorating hypertension and improving or enhancing motor function. [Brief explanation of the drawings]

[0011] [Figure 1]Effect of potassium nitrate and sugar alcohols in combination on Neisseria mucosa in dental plaque biofilm flora. [Figure 2] Effect of potassium nitrate and sugar alcohols in combination on Neisseria bacteria in dental plaque biofilm flora. [Figure 3] Dose-dependent effect of combined potassium nitrate and erythritol on Neisseria mucosa in dental plaque biofilm flora. [Figure 4] The effect of combined potassium nitrate and erythritol on Neisseria species in dental plaque biofilm flora (dose-dependent). DETAILED DESCRIPTION OF THE INVENTION

[0012] In the present invention, the nitrate ion supplying compound includes a compound capable of generating nitrate ions when contacted with water, and is preferably a nitrate salt. Examples of the nitrate salt include alkali metal nitrate salts and alkaline earth metal nitrate salts, and specific examples include sodium nitrate, potassium nitrate, and calcium nitrate, with potassium nitrate being preferred. The nitrate salt is preferably of synthetic origin, but may also be of natural origin.

[0013] In the present invention, the sugar alcohol may be one or more selected from erythritol, xylitol, and mannitol. From the viewpoint of increasing the content of Neisseria mucosa in the oral cavity, it is preferable to use at least erythritol as the sugar alcohol.

[0014] The nitrate ion-supplying compound and sugar alcohol are used in combination, and the form of the combination may be a formulation in which effective amounts of each compound are formulated in a suitable ratio into a single dosage form as a combination drug, or a kit in which drugs containing effective amounts of each compound are formulated separately and can be used simultaneously or separately at intervals. A combination drug form is preferred.

[0015] In the present invention, the ratio of the combination of the nitrate ion-donating compound and the sugar alcohol can be appropriately selected, but for example, the sugar alcohol is preferably 2.5 parts by mass or more, more preferably 5 parts by mass or more, more preferably 10 parts by mass or more, and preferably 40 parts by mass or less, more preferably 30 parts by mass or less, more preferably 20 parts by mass or less, relative to 1 part by mass of the nitrate ion-donating compound. Also, the ratio is preferably 2.5 to 40 parts by mass, more preferably 5 to 20 parts by mass, more preferably 10 to 20 parts by mass.

[0016] As shown in the Examples below, a dental plaque suspension containing plaque collected from healthy individuals was cultured in a medium containing potassium nitrate and various sugar alcohols (erythritol, xylitol, mannitol) alone or in combination to produce a dental plaque biofilm, and the content of Neisseria mucosa and Neisseria spp. was calculated. When potassium nitrate or various sugar alcohols alone, or potassium nitrate in combination with sorbitol or maltitol, no significant changes were observed in the content of Neisseria mucosa and the content of Neisseria spp. compared to the control group, however, when potassium nitrate was used in combination with erythritol, xylitol, or mannitol, the content of Neisseria mucosa and the content of Neisseria spp. increased significantly. Therefore, a combination of a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol can be an oral Neisseria mucosa-increasing agent, an oral Neisseria spp.-increasing agent, or an oral flora-improving agent, and can be used to produce an oral Neisseria mucosa-increasing agent, an oral Neisseria spp.-increasing agent, or an oral flora-improving agent. Furthermore, a combination of a nitrate ion-receiving compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol can be used to increase oral Neisseria mucosa, increase oral Neisseria bacteria, or improve oral flora. The use can be administered to humans or non-human animals, or to specimens derived therefrom, and can be therapeutic or non-therapeutic. Non-human animals include non-human mammals, such as apes, other primates, mice, rats, horses, cows, pigs, sheep, dogs, cats, hamsters, and companion animals.

[0017] In the present invention, Neisseria mucosa is a gram-negative cocci belonging to the genus Neisseria in the family Neisseria, and is a non-pathogenic normal bacterium inhabiting the oral cavity. The genus Neisseria refers to non-pathogenic oral bacteria belonging to the genus Neisseria in the family Neisseria, and includes Neisseria mucosa, as well as Neisseria sicca, Neisseria flava, Neisseria subflava, Neisseria flavescens, and Neisseria elongata.

[0018] In the present invention, "increasing oral Neisseria mucosa" means increasing the content of Neisseria mucosa in the oral cavity by, for example, activating the metabolism of at least Neisseria mucosa among oral Neisseria bacteria. Also, "increasing oral Neisseria bacteria" means increasing oral Neisseria bacteria. In addition, "improvement of oral bacterial flora" refers to an increase in the content of oral Neisseria mucosa or oral Neisseria spp., the physiological functions of Neisseria mucosa or oral Neisseria spp. being exerted more effectively, and the oral bacterial flora changing in a direction that is favorable for the host.

[0019] The Neisseria mucosa content can be calculated, for example, by measuring the copy number of the nitrate reductase gene (narG) of Neisseria mucosa using real-time PCR as the target gene, and calculating the ratio to the copy number of the rRNA16S gene of all bacteria, as shown in the examples described below. In addition, the content of Neisseria bacteria can be calculated by measuring the copy number of the rRNA16S gene by real-time PCR using a probe that specifically detects the rRNA16S gene of all Neisseria species, and calculating the ratio to the copy number of the rRNA16S gene of all bacteria.

[0020] As mentioned above, Neisseria mucosa is useful for preventing or treating periodontal disease because its bacterial cells or cultures thereof suppress infection of gingival cells by periodontal pathogens and suppress gingival inflammation (Patent Document 1). Periodontal pathogens refer to bacteria such as Porphyromonas gingivalis and Fusobacterium nucleatum that infect periodontal tissues and cause gingivitis and even periodontal tissue destruction. In addition, Neisseria bacteria produce nitrate ions (NO3 - ) to nitrite ions (NO2 - Since nitrate ion-supplying compounds have the ability to reduce nitrate ions to nitrite and nitric oxide (NO) (Non-Patent Document 1), they are thought to be reduced by Neisseria bacteria to nitrite and then converted into nitrogen oxides, including nitric oxide, which exert a blood pressure lowering effect due to improved vascular function and an exercise function improving effect due to improved skeletal muscle contractility (Non-Patent Documents 2, 3, and 4). Therefore, the oral Neisseria mucosa-increasing agent, Neisseria bacteria-increasing agent, or oral bacterial flora-improving agent of the present invention, i.e., a combination of a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol, serves as a nitric oxide-increasing agent, an agent for preventing or improving high blood pressure, an agent for improving motor function, and an agent for preventing or improving periodontal disease, and can be used to increase nitric oxide, improve or improve motor function, prevent or improve high blood pressure, and prevent or improve periodontal disease. Here, the agent for preventing or improving periodontal disease exerts its effect by inhibiting the infection of gingival cells by periodontal pathogens caused by Neisseria mucosa, and therefore the amount of bactericide used can be reduced, making it possible to produce a safer formulation.

[0021] Here, "increasing nitric oxide" means increasing the natural concentration of nitric oxide in the body. Increasing nitric oxide makes it possible to prevent or improve vascular dysfunction caused by nitric oxide deficiency (e.g., vascular endothelial dysfunction, arteriosclerosis, etc.) and the associated pathologies (hypertension, myocardial infarction, cerebral infarction, etc.). Increasing nitric oxide also improves skeletal muscle contractility, thereby improving or enhancing motor function. "Periodontal disease" refers to an inflammatory disease in which inflammation of the gums is caused by bacteria in dental plaque, and includes gingivitis and periodontitis. Additionally, "prevention" refers to preventing or delaying the onset of a disease or condition in an individual, or reducing the risk of an individual developing a disease or condition. Furthermore, "improvement" means improvement of a disease, symptom or condition, prevention or delay of worsening of a disease, symptom or condition, or reversal, prevention or delay of progression of a disease or symptom, and includes the concept of so-called "treatment." "Improvement" refers to a physical function moving in a better or more advanced direction from the current state.

[0022] The agent for increasing oral Neisseria mucosa, agent for increasing Neisseria spp., agent for improving oral bacterial flora, agent for preventing or improving periodontal disease, agent for increasing nitric oxide, agent for preventing or improving hypertension, and agent for improving or enhancing motor function of the present invention can be used by itself as a pharmaceutical, quasi-drug, food, or oral composition for increasing oral Neisseria mucosa, increasing oral Neisseria spp., improving oral bacterial flora, preventing or improving periodontal disease, increasing nitric oxide, preventing or improving hypertension, or improving or enhancing motor function, or can be a material or preparation to be incorporated into such pharmaceutical, quasi-drug, food, or oral composition. In addition, foods include foods that are based on concepts such as increasing oral Neisseria mucosa, increasing oral Neisseria bacteria, improving oral flora, suppressing periodontal pathogen infection, preventing or improving periodontal disease, increasing nitric oxide, preventing or improving high blood pressure, improving or enhancing motor function, etc., and are labeled as such as functional foods, foods for specified health uses, foods for the sick, and supplements.

[0023] When used as a pharmaceutical or quasi-drug, the formulation is preferably an oral administration form, and examples of the dosage form include liquid preparations; solid preparations such as tablets, granules, fine granules, powders, and tablets; and various forms such as capsules, oral sprays, and troches containing the liquid preparations or solid preparations.

[0024] When used as a food, the forms include beverages such as fruit or vegetable juice drinks, carbonated drinks, tea drinks, dairy drinks, fermented milk, fermented fruit juice, fermented vegetable juice, alcoholic drinks, and soft drinks, as well as various foods such as jelly foods, various snacks, baked goods, cakes, chocolate, jam, bread, gum, candy, soups, pickles, and tsukudani (foods boiled in soy sauce), as well as supplements in the same form as the oral administration formulations described above (tablets, capsules, syrup, etc.).

[0025] Specific forms for use as oral compositions include mouthwash, mouthwash, toothpaste, powder toothpaste, dentifrice, oral ointment, gel, tablet, granule, fine granule, gummy jelly, troche, tablet, capsule, candy, chewing gum, etc., and preferred are toothpaste, mouthwash, gummy jelly, and troche.

[0026] The pharmaceuticals, quasi-drugs, foods, and oral compositions can be prepared according to standard methods by appropriately combining a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol with other active ingredients, or additives acceptable for pharmaceuticals, foods, etc., or any food ingredients, etc.

[0027] The content of the nitrate ion-supplying compound and sugar alcohol in the above-mentioned pharmaceuticals, quasi-drugs, foods, and oral compositions is not particularly limited, and may be adjusted appropriately according to the daily dosage, etc.

[0028] The intake or administration amount of the above-mentioned pharmaceuticals, quasi-drugs, foods, and oral compositions can be determined appropriately depending on various conditions such as the patient's weight, age, sex, and symptoms, but for an adult, the daily amount of nitrate ion-supplying compounds is 60 mg or more, preferably 200 mg or more, more preferably 600 mg or more, and 6000 mg or less, preferably 2000 mg or less, and more preferably 1000 mg or less. Also, the daily amount of sugar alcohols is 300 mg or more, preferably 1000 mg or more, more preferably 3000 mg or more, and 30000 mg or less, preferably 10000 mg or less, and more preferably 5000 mg or less.

[0029] The oral Neisseria mucosa-increasing agent, oral Neisseria bacteria-increasing agent, oral bacterial flora improving agent, periodontal pathogen infection inhibitor, periodontal disease prevention or amelioration agent, nitric oxide increasing agent, hypertension prevention or amelioration agent, and motor function improving or enhancing agent of the present invention are preferably applicable to humans infected with periodontal pathogens, humans suffering from gingival inflammation such as gingivitis or periodontitis, humans wishing to prevent the onset of gingival inflammation, humans with hypertension, humans wishing to lower their blood pressure, humans wishing to improve or enhance motor function, etc.

[0030] In relation to the above-described embodiment, the present invention further discloses the following aspects. <1> An oral Neisseria mucosa increasing agent containing as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. <2> An agent for increasing the number of Neisseria bacteria in the oral cavity, which comprises as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. <3> An oral flora improving agent containing, as active ingredients, a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. <4> A periodontal pathogen infection inhibitor comprising a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol as active ingredients. <5> An agent for preventing or ameliorating periodontal disease, comprising as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. <6> A nitric oxide enhancer comprising a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol as active ingredients. <7> An agent for preventing or improving hypertension, comprising as active ingredients a nitrate ion-donating compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. <8> An agent for improving or enhancing motor function, comprising as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol.

[0031] <9> A food for increasing Neisseria mucosa in the oral cavity, the food containing as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. <10> A food for increasing the number of Neisseria bacteria in the oral cavity, which contains as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. <11> A food for improving oral flora, which contains as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. <12> A food for inhibiting periodontal pathogen infection, comprising as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. <13> A food for preventing or improving periodontal disease, which contains as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. <14> A food for increasing nitric oxide, which contains as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. <15> A food for preventing or improving hypertension, which contains as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol. <16> A food for improving or enhancing motor function, which contains as active ingredients a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol.

[0032] <17> Use of a combination of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol for producing an oral Neisseria mucosa-increasing agent or a food for increasing oral Neisseria mucosa. <18> 1. Use of a combination of a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol for producing an agent for increasing oral Neisseria bacteria or a food for increasing oral Neisseria bacteria. <19> 1. Use of a combination of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol for producing an oral flora improver or a food for improving oral flora. <20> 1. Use of a combination of a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol for producing an agent for inhibiting periodontal pathogen infection or a food for inhibiting periodontal pathogen infection. <21> 1. Use of a combination of a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol for producing an agent for preventing or ameliorating periodontal disease or a food for preventing or ameliorating periodontal disease. <22> 1. Use of a combination of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol for producing a nitric oxide enhancer or a food for increasing nitric oxide. <23> 1. Use of a combination of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol for producing an agent for preventing or ameliorating hypertension or a food for preventing or ameliorating hypertension. <24> 1. Use of a combination of a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol for producing an agent for improving or enhancing motor function or a food for improving or enhancing motor function.

[0033] <25> A combination of a nitrate ion-donating compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol for use in increasing oral Neisseria mucosa. <26> A combination of a nitrate ion-donating compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol for use in increasing oral Neisseria bacteria. <27> A combination of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol for use in improving oral bacterial flora. <28> A combination of a nitrate ion-donating compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol for use in inhibiting periodontal pathogen infection. <29> A combination of a nitrate ion-donating compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol for use in preventing or ameliorating periodontal disease. <30 A combination of a nitrate ion donating compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol for use in increasing nitric oxide. <31> A combination of a nitrate ion-donating compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol for use in preventing or improving hypertension. <32> A combination of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol for use in improving or enhancing motor function.

[0034] <33> Non-therapeutic use of a combination of a nitrate ion-donating compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to increase Neisseria mucosa in the oral cavity. <34> 1. Non-therapeutic use of a combination of a nitrate ion-providing compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to increase oral Neisseria bacteria. <35> 1. Non-therapeutic use of a combination of a nitrate ion-donating compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol for improving oral flora. <36> A non-therapeutic use of a combination of a nitrate ion-donating compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol for suppressing periodontal pathogen infection. <37> 1. Non-therapeutic use of a combination of a nitrate ion-donating compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol for preventing or ameliorating periodontal disease. <38> 1. Non-therapeutic use of a combination of a nitrate ion-donating compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol to increase nitric oxide. <39> 1. Non-therapeutic use of a combination of a nitrate ion-donating compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol for preventing or ameliorating high blood pressure. <40> 1. Non-therapeutic use of a combination of a nitrate ion-donating compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol to improve or enhance motor function.

[0035] <41> A method for increasing Neisseria mucosa in the oral cavity, comprising administering or ingesting an effective amount of a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol and mannitol to a subject in need thereof. <42> A method for increasing Neisseria bacteria in the oral cavity, comprising administering or ingesting an effective amount of a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to a subject in need thereof. <43> A method for improving oral bacterial flora, comprising administering or ingesting an effective amount of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to a subject in need thereof. <44> A method for inhibiting periodontal pathogen infection, comprising administering or ingesting an effective amount of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to a subject in need thereof. <45> A method for preventing or ameliorating periodontal disease, comprising administering or ingesting an effective amount of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to a subject in need thereof. <46> A method for increasing nitric oxide, comprising administering or ingesting an effective amount of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to a subject in need thereof. <47> A method for preventing or ameliorating hypertension, comprising administering or ingesting an effective amount of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to a subject in need thereof. <48> A method for improving or enhancing motor function, comprising administering or ingesting an effective amount of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to a subject in need thereof. <49> <1> ~ <48> In the above, the sugar alcohol is preferably erythritol, or erythritol and xylitol, or erythritol and mannitol, or erythritol, xylitol and mannitol. <50> <1> ~ <49> In the above, the nitrate ion supplying compound is preferably a nitrate salt, more preferably sodium nitrate or potassium nitrate. <51> <1> ~ <50> In the above, the ratio of the combination of the nitrate ion supplying compound and the sugar alcohol is preferably 2.5 parts by mass or more, more preferably 5 parts by mass or more, more preferably 10 parts by mass or more, and preferably 40 parts by mass or less, more preferably 30 parts by mass or less, more preferably 20 parts by mass or less, per part by mass of the nitrate ion supplying compound, and is also preferably 2.5 to 40 parts by mass, more preferably 5 to 20 parts by mass, more preferably 10 to 20 parts by mass. <52> <1> ~ <50> In this case, the intake or administration amount of the nitrate ion-supplying compound and sugar alcohol per adult per day is 60 mg or more, preferably 200 mg or more, more preferably 600 mg or more, and 6000 mg or less, preferably 2000 mg or less, and more preferably 2000 mg or less, of the nitrate ion-supplying compound, and 300 mg or more, preferably 1000 mg or more, more preferably 3000 mg or more, and 30000 mg or less, preferably 10000 mg or less, and more preferably 5000 mg or less of the sugar alcohol. <53> <3> , <11> , <19> , <27> or <35> In this study, improvement of oral flora is an increase in the content of oral Neisseria mucosa or oral Neisseria spp. [Example]

[0036] Effect of combined administration of nitrate and sugar alcohols on Neisseria mucosa in dental plaque biofilm flora (1) Plaque collection and pretreatment Plaque adhering to the tooth surface of a healthy male in his twenties 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 1Plus (Juji Field). The prepared plaque suspension was stored at -80°C until use.

[0037] (2) Preparation of a dental plaque biofilm model using dental plaque suspension culture A culture medium containing plaque bacteria was prepared by diluting the dental plaque suspension 1000-fold with Brain Heart Infusion (BHI) medium (BD Japan) containing the test compounds potassium nitrate (Wako), erythritol (Wako), xylitol (Wako), mannitol (Wako), sorbitol (Wako), and maltitol (Wako). One milliliter 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 Anaeropack Bikouki (Mitsubishi Gas Chemical) for 48 hours to produce a dental plaque biofilm. After 48 hours of culture, the supernatant was carefully aspirated, taking care not to disrupt the plaque biofilm on the bottom, to prepare a dental plaque biofilm sample.

[0038] (3) Extraction of genomic DNA from dental plaque biofilm A 200 μL / well solution of 20 mg / mL lysozyme (WAKO) (containing 20 mM Tris-HCl (pH 8.0), 2 mM EDTA, and 1.2% Triton X-100) was added to the dental plaque biofilm sample attached to the bottom of the plate, and the plate was incubated at 37°C with shaking (160 rpm). Genomic DNA was extracted using the Dneasy Blood & Tissue kit (QIAGEN) according to the kit's standard protocol. Finally, the sample was eluted with 300 μL of elution solution to prepare the genomic DNA sample.

[0039] (4) Quantification of Neisseria mucosa and Neisseria spp. in genomic DNA samples Neisseria mucosa and Neisseria species were quantified by real-time PCR using TaqMan probes. To specifically detect Neisseria mucosa, only Neisseria mucosa possesses the nitrate reductase gene (narG) (see Non-Patent Document 1). Therefore, TaqMan probes and primers were prepared using narG as the target gene (Table 1). TaqMan probes and primers were also prepared to specifically detect the rRNA16S genes of all Neisseria species (Table 1). TaqMan probes and primers to detect the rRNA16S genes of all bacteria were prepared using the sequences previously described (Nadkarni, MA, et al., Microbiology (2002) 148, 257-266) (Table 1). The composition of the reaction solution and reaction conditions for real-time PCR are shown in Table 2.

[0040] [Table 1]

[0041] [Table 2]

[0042] A calibration curve was created using PCR products with known copy numbers as standards, and the copy numbers of the Neisseria mucosa nitrate reductase gene (narG) and the rRNA16S gene of Neisseria spp., as well as the copy numbers of the rRNA16S gene of all bacteria, were calculated. The Neisseria mucosa content and the Neisseria spp. content were calculated using the following formula.

[0043]

number

[0044] (5) Results The effects of 1% (W / V) potassium nitrate and 5% (W / V) sugar alcohols (erythritol, xylitol, mannitol, sorbitol, and maltitol) on Neisseria mucosa and Neisseria spp. in dental plaque biofilm flora are shown in Figures 1 and 2. As shown in Figures 1 and 2, potassium nitrate, erythritol, xylitol, mannitol, sorbitol, and maltitol alone did not have a significant effect on the Neisseria mucosa content or the Neisseria spp. content compared to the control group, and the combination of potassium nitrate with sorbitol or maltitol did not have a significant effect either. However, the combination of potassium nitrate with erythritol, xylitol, or mannitol significantly increased the Neisseria mucosa content and the Neisseria spp. content. Furthermore, the dose dependency of the effect of combined use of potassium nitrate and erythritol on increasing the content of Neisseria mucosa and Neisseria spp. is shown in Figures 3 and 4. As shown in Figures 3 and 4, potassium nitrate alone showed a weak effect of increasing the content of Neisseria mucosa and Neisseria spp. only at 0.2% potassium nitrate, but when combined with erythritol, a significant increase in the content of Neisseria mucosa and Neisseria spp. was observed in an erythritol dose-dependent manner.

Claims

1. An agent for increasing the amount of Neisseria mucosa in the oral cavity, which comprises a nitrate ion supplying compound and one or more sugar alcohols selected from xylitol and mannitol as active ingredients.

2. An agent for increasing the number of Neisseria bacteria in the oral cavity, which comprises a nitrate ion supplying compound and one or more sugar alcohols selected from xylitol and mannitol as active ingredients.

3. An oral flora improving agent comprising, as active ingredients, a nitrate ion supplying compound and one or more sugar alcohols selected from xylitol and mannitol.

4. The oral flora improving agent according to claim 3, wherein the improvement of the oral flora is an increase in the content of Neisseria mucosa or Neisseria bacteria in the oral cavity.

5. A periodontal pathogen infection inhibitor comprising a nitrate ion supplying compound and one or more sugar alcohols selected from xylitol and mannitol as active ingredients.

6. An agent for preventing or ameliorating periodontal disease, comprising as active ingredients a nitrate ion-donating compound and one or more sugar alcohols selected from xylitol and mannitol.

7. The agent according to any one of claims 1 to 6, wherein the nitrate ion-donating compound is a nitrate salt.

8. 8. The agent according to claim 7, wherein the nitrate is sodium nitrate or potassium nitrate.

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