Oral Neisseria mucosa increasing agent

Abstract

This material provides a useful component for increasing the content of Neisseria mucosa in the oral cavity. [Solution] An oral Neisseria mucosa increasing agent comprising a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients.

Description

Technical Field

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

Background Art

[0002] In the human body, microorganisms construct and inhabit a commensal flora based on complex interactions in various sites. The composition of the commensal flora varies among individuals, and the relationship between this individual difference and the host's health status has been pointed out many times in recent years and has attracted attention. The oral cavity, like the intestine, is a microbial ecosystem in which a wide variety of bacteria coexist. Although several pathogenic bacteria have been identified for oral infectious diseases such as dental caries and periodontal diseases, these bacteria also coexist in the oral cavity as members of the normal ecosystem. Therefore, it is presumed that the interaction with other commensal bacteria surrounding the onset of the disease plays a major role. Conventionally, in the prevention and treatment of oral diseases, the removal of pathogenic bacteria that directly cause the disease has been the goal, but it is considered that the induction of a commensal flora in which pathogenic bacteria are less likely to thrive can also be an approach in the prevention of oral diseases.

[0003] Neisseria mucosa is known to exist as a non-pathogenic commensal bacterium in the oral cavity, together with Neisseria species such as Neisseria sicca, Neisseria flava, Neisseria subflava, Neisseria flavescens, and Neisseria elongata. The present applicant has recently found that the cells or cultures of Neisseria mucosa inhibit the infection of periodontal pathogenic bacteria to gingival cells and are useful for the prevention or treatment of periodontal disease (Patent Document 1). In addition, Neisseria bacteria possess nitrate reductase and nitrite reductase and have the ability to reduce nitrate ions (NO 3 - ) to nitrite ions (NO2 - ) or nitric oxide (NO). It has been reported (Non-Patent Literature 1). When nitrate ions are ingested orally, it is thought that a portion of the nitrate ions absorbed into the body migrate from the blood to the saliva in high concentrations, where they are reduced to nitrite by nitrate-reducing bacteria that reside in the oral cavity, and then enzymatically or non-enzymatically produce nitric oxide (NO) in the stomach and various tissues. Nitric oxide is an important compound that exerts various effects throughout the body, including intercellular communication, antibacterial activity, defense against pathogens, vasodilation, and effects on muscle contractility. In particular, it is expected to have antibacterial activity against periodontal pathogens, as well as blood pressure reduction based on improved vascular function and improvement or enhancement of motor function based on improved muscle contractility (Non-Patent Literature 2-5). Therefore, increasing the oral cavity content of Neisseria bacteria is beneficial.

[0004] On the other hand, potassium ions have the effect of reducing the hypersensitivity of dental pulp nerves, and potassium nitrate and other substances are known to be effective against tooth sensitivity (Non-Patent Literature 6). Furthermore, many sugar alcohols have the effect of promoting saliva secretion and tooth remineralization, and erythritol, xylitol, and sorbitol have been reported to have a caries-preventive effect (Non-Patent Literature 7). Additionally, sugar alcohols such as erythritol have been reported to inhibit biofilm formation (Patent Literature 2). However, the effects of potassium nitrate and sugar alcohols on Neisseria bacteria in the oral cavity are completely unknown. [Prior art documents] [Patent Documents]

[0005] [Patent Document 1] Japanese Patent Application No. 2020-200517 [Patent Document 2] Japanese Patent Publication 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] Katsumi Shibata, Agriculture and Science, April 2019, pp. 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. [Overview of the project] [Problems that the invention aims to solve]

[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 found that by combining a nitrate ion supply compound with a specific sugar alcohol, the content of Neisseria mucosa and Neisseria species in the oral cavity can be increased, which is useful for increasing nitric oxide and preventing or treating periodontal disease.

[0009] In other words, the present invention relates to the following 1) to 8). 1) An oral Neisseria mucosa increasing agent comprising a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. 2) An oral Neisseria bacteria increasing agent comprising a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. 3) An oral microbiome improving agent comprising a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. 4) A periodontal pathogen infection inhibitor comprising a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. 5) A preventive or ameliorative agent for periodontal disease, comprising a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. 6) A nitric oxide increasing agent comprising a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. 7) A hypertension preventive or ameliorative agent comprising a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. 8) An agent for improving or enhancing athletic performance, comprising a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. [Effects of the Invention]

[0010] According to the present invention, the content of Neisseria mucosa and Neisseria species in the oral cavity can be increased, making it possible to prevent or treat periodontal disease. Furthermore, by supplying nitric oxide, it is possible to improve vascular function and skeletal muscle contractility, making it possible to prevent or improve hypertension and improve or enhance motor function. [Brief explanation of the drawing]

[0011] [Figure 1]Effect of the combined use of potassium nitrate and each sugar alcohol on Neisseria mucosa in dental plaque biofilm flora. [Figure 2] Effect of the combined use of potassium nitrate and each sugar alcohol on Neisseria spp. in dental plaque biofilm flora. [Figure 3] Effect of the combined use of potassium nitrate and erythritol on Neisseria mucosa in dental plaque biofilm flora (dose-dependence). [Figure 4] Effect of the combined use of potassium nitrate and erythritol on Neisseria spp. in dental plaque biofilm flora (dose-dependence).

Mode for Carrying Out the Invention

[0012] In the present invention, examples of the nitrate ion supply compound include compounds capable of generating nitrate ions when contacted with water, and nitrates are preferred. Examples of nitrates include alkali metal nitrates and alkaline earth metal nitrates. Specific examples include sodium nitrate, potassium nitrate, calcium nitrate, etc., and potassium nitrate is preferred. Nitrates are preferably of synthetic origin, but those of natural origin may also be used.

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

[0014] Such a nitrate ion supply compound and sugar alcohol are used in combination. The form may be a formulation in which each effective amount is formulated into one dosage form at an appropriate mixing ratio as a formulation, or a kit in which drugs containing each effective amount are separately formulated and can be used simultaneously or separately at intervals. Preferably, it is in the form of a formulation.

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

[0016] As shown in the examples below, when a 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 plaque biofilm, and the content of Neisseria mucosa and Neisseria species was calculated, no significant change in the content of Neisseria mucosa and Neisseria species was observed compared to the control group when potassium nitrate or various sugar alcohols were used alone, or when potassium nitrate was used in combination with sorbitol or maltitol. However, when potassium nitrate was used in combination with erythritol, xylitol, or mannitol, the content of Neisseria mucosa and Neisseria species increased significantly. Therefore, a combination of a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol can serve as an oral Neisseria mucosa increasing agent, an oral Neisseria bacteria increasing agent, or an oral microbiota improving agent, and can be used to manufacture oral Neisseria mucosa increasing agents, oral Neisseria bacteria increasing agents, or oral microbiota improving agents. Furthermore, combinations of nitrate ion-supplying compounds with one or more sugar alcohols selected from erythritol, xylitol, and mannitol can be used to increase oral Neisseria mucosa, increase oral Neisseria species, or improve the oral microbiota. Here, such use may be administration to humans or non-human animals, or use in specimens derived therefrom, and may be therapeutic or non-therapeutic. Here, non-human animals include non-human mammals such as apes, other primates, mice, rats, horses, cattle, pigs, sheep, dogs, cats, hamsters, and companion animals.

[0017] In this invention, Neisseria mucosa is a Gram-negative cocci belonging to the genus Neisseria of the family Neisseriaceae, and is a non-pathogenic commensal bacterium of the oral cavity. Furthermore, the genus Neisseria refers to non-pathogenic oral commensal bacteria belonging to the genus Neisseria of the family Neisseriaceae, and includes not only Neisseria mucosa, but also Neisseria sicca, Neisseria flava, Neisseria subflava, Neisseria flavescens, Neisseria elongata, and others.

[0018] In this invention, "increase in oral Neisseria mucosa" means an increase in the content of Neisseria mucosa in the oral cavity, such as by activating the metabolism of at least Neisseria mucosa among oral Neisseria bacteria. Furthermore, "increase in oral Neisseria bacteria" means an increase in oral Neisseria bacteria. Furthermore, "improvement of oral microbiota" refers to an increase in the content of Neisseria mucosa or Neisseria species in the oral cavity, the more effectively the physiological functions of Neisseria mucosa or Neisseria species in the oral cavity are exerted, and a change in the oral microbiota that is beneficial to the host.

[0019] Furthermore, the Neisseria mucosa content can be calculated, for example, by using the Neisseria mucosa nitrate reductase gene (narG) as the target gene, measuring the copy number of the gene by real-time PCR, and determining the ratio to the copy number of the rRNA16S gene of all bacteria, as shown in the example described later. Furthermore, the Neisseria content can be calculated by measuring the copy number of the rRNA16S gene using a real-time PCR method with a probe that specifically detects the rRNA16S gene of all Neisseria species, and then determining the ratio to the total copy number of the rRNA16S gene of all bacteria.

[0020] As mentioned above, Neisseria mucosa is useful in the prevention or treatment of periodontal disease because its cells or cultures suppress the infection of gingival cells by periodontal pathogens and thus suppress gingival inflammation (see 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. Furthermore, Neisseria bacteria produce nitrate ions (NO3 - ) nitrite ions (NO2 - Because it has the ability to reduce nitrate ions to nitrites and nitric oxide (NO) (as mentioned in Non-Patent Document 1), it is thought that nitrate ion supply compounds are reduced to nitrites by Neisseria bacteria, and further converted to nitrogen oxides containing nitric oxide, thereby exerting blood pressure lowering effects based on improved vascular function and improved exercise function effects based on improved skeletal muscle contractility (as mentioned in Non-Patent Documents 2, 3, and 4). Therefore, the present invention provides an oral Neisseria mucosa increasing agent, a Neisseria species increasing agent, or an oral microbiota improving agent, that is, a combination of a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol, which can be used to increase nitric oxide, prevent or improve hypertension, improve athletic performance, and prevent or improve periodontal disease. Here, since periodontal disease prevention or improvement agents exert their effects through the inhibitory action of Neisseria mucosa on the infection of gingival cells by periodontal pathogens, the amount of bactericide used can be reduced, potentially resulting in a safer formulation.

[0021] Here, "increase in nitric oxide" means increasing the naturally occurring nitric oxide concentration in the body. By increasing nitric oxide, it is possible to prevent or improve the decline in vascular function (e.g., impaired endothelial function, arteriosclerosis, etc.) and the resulting pathological conditions (hypertension, myocardial infarction, cerebral infarction, etc.) caused by nitric oxide deficiency. In addition, an increase in nitric oxide can improve the contractility of skeletal muscles and improve or enhance motor function. "Periodontal disease" refers to an inflammatory disease in which bacteria in plaque cause inflammation of the gums, and it encompasses both gingivitis and periodontitis. Furthermore, "prevention" refers to preventing or delaying the onset of a disease or symptom in an individual, or reducing the risk of an individual developing a disease or symptom. Furthermore, "improvement" means a rise in the disease, symptoms, or condition, prevention or delay of the worsening of the disease, symptoms, or condition, or a reversal, prevention, or delay of the progression of the disease or symptoms, and encompasses the concept of so-called "treatment." Furthermore, "improvement" refers to a movement towards a better or superior state of physical function, etc., from the current point in time.

[0022] The present invention provides an oral Neisseria mucosa increasing agent, a Neisseria species increasing agent, an oral microbiota improving agent, a periodontal disease prevention or improvement agent, a nitric oxide increasing agent, a hypertension prevention or improvement agent, and an exercise function improvement or enhancement agent, which can themselves serve as pharmaceuticals, quasi-drugs, foods, or oral compositions for increasing oral Neisseria mucosa, increasing oral Neisseria species, improving oral microbiota, preventing or improving periodontal disease, increasing nitric oxide, preventing or improving hypertension, or improving or enhancing exercise function. They can also be materials or formulations used when incorporated into such pharmaceuticals, quasi-drugs, foods, or oral compositions. Furthermore, the term "food" includes foods, functional foods, foods for specified health uses, foods for sick people, and supplements that are based on the concepts of increasing oral Neisseria mucosa, increasing oral Neisseria bacteria, improving oral microbiota, suppressing periodontal pathogen infection, preventing or improving periodontal disease, increasing nitric oxide, preventing or improving hypertension, improving or enhancing motor function, etc., and that display such claims as appropriate.

[0023] When used as a pharmaceutical or quasi-drug, the preferred form is for oral administration. The dosage forms include, for example, liquid preparations; solid preparations such as tablets, granules, fine granules, powders, or tablets; or various forms such as capsules, oral sprays, and lozenges containing the liquid or solid preparation.

[0024] When used as a food product, it can take the form of beverages such as fruit or vegetable juices, carbonated drinks, tea-based beverages, milk beverages, fermented milk, fermented fruit juices, fermented vegetable juices, alcoholic beverages, and soft drinks, as well as various foods such as jelly-like foods, various snacks, baked goods, cakes, chocolates, jams, bread, gum, candies, soups, pickles, and preserved foods, and supplements in the same form as the oral administration preparations mentioned above (tablets, capsules, syrups, etc.).

[0025] Specific forms in which the oral composition can be used include mouthwash, toothpaste, toothpaste powder, toothpaste liquid, oral ointment, gel, tablet, granule, fine granule, gummy candy, lozenge, capsule, candy, chewing gum, etc., with toothpaste, mouthwash, gummy candy, and lozenge being preferred.

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

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

[0028] In the above-mentioned pharmaceuticals, quasi-drugs, foods, and oral compositions, the intake or dosage can be appropriately determined according to various conditions such as the patient's weight, age, sex, and symptoms. However, for an adult, a daily dose of nitrate ion supplying compound 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. In addition, a sugar alcohol dose of 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 present invention's oral Neisseria mucosa increasing agent, oral Neisseria species increasing agent, oral microbiota improving agent, periodontal pathogen infection inhibitor, periodontal disease prevention or improvement agent, nitric oxide increasing agent, hypertension prevention or improvement agent, and motor function improvement or enhancement agent are preferably intended for use by people infected with periodontal pathogens, people suffering from gingivitis such as gingivitis or periodontitis, people who wish to prevent the onset of gingivitis, people with hypertension, people who wish to lower their blood pressure, and people who wish to improve or enhance their motor function.

[0030] With regard to the embodiments described above, the present invention further discloses the following embodiments. <1> An oral Neisseria mucosa increasing agent comprising a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. <2> An oral Neisseria bacteria-promoting agent containing a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. <3> An oral microbiome improving agent containing a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. <4> A periodontal pathogen infection inhibitor containing a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. <5> A periodontal disease prevention or improvement agent comprising a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. <6> A nitric oxide increasing agent comprising a nitrate ion supply 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 a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. <8> An agent for improving or enhancing athletic performance, comprising a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients.

[0031] <9> A food product for increasing Neisseria mucosa in the oral cavity, containing a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. <10> A food product for increasing Neisseria bacteria in the oral cavity, containing a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. <11> A food product for improving oral microbiota, containing a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. <12> A food product for suppressing periodontal disease-causing bacteria infection, containing a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. <13> A food product for the prevention or improvement of periodontal disease, containing a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. <14> A food product for increasing nitric oxide, containing a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. <15> A food product for preventing or improving hypertension, containing a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients. <16> A food product for improving or enhancing athletic performance, containing a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients.

[0032] <17> The use of a combination of a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol for the manufacture of oral Neisseria mucosa increasing agents or oral Neisseria mucosa increasing foods. <18> The use of a combination of a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol for the production of an oral Neisseria bacteria increasing agent or oral Neisseria bacteria increasing food. <19> The use of a combination of a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol for the manufacture of an oral microbiome improving agent or a food for improving the oral microbiome. <20> The use of a combination of a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol for the manufacture of a periodontal pathogen infection inhibitor or a food for inhibiting periodontal pathogen infection. <21> The use of a combination of a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol for the manufacture of a periodontal disease prevention or improvement agent or a food for the prevention or improvement of periodontal disease. <22> The use of a combination of a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol for the production of a nitric oxide increasing agent or a food for increasing nitric oxide. <23> The use of a combination of a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol for the manufacture of an agent for preventing or improving hypertension or a food for preventing or improving hypertension. <24> The use of a combination of a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol for the manufacture of an agent for improving or enhancing athletic function or a food for improving or enhancing athletic function.

[0033] <25> A combination of a nitrate ion supply 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-supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol, for use in increasing Neisseria bacteria in the oral cavity. <27> A combination of a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol, for use in improving the oral microbiome. <28> A combination of a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol, for use in suppressing periodontal pathogen infection. <29> A combination of a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol, for use in the prevention or improvement of periodontal disease. <30 A combination of a nitrate ion supply 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-supplying 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 supply 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 supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to increase oral Neisseria mucosa. <34> Non-therapeutic use of a combination of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to increase Neisseria bacteria in the oral cavity. <35> Non-therapeutic use of a combination of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to improve the oral microbiome. <36> Non-therapeutic use of a combination of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to suppress periodontal pathogen infection. <37> Non-therapeutic use of a combination of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol for the prevention or improvement of periodontal disease. <38> Non-therapeutic use of a combination of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to increase nitric oxide. <39> Non-therapeutic use of a combination of a nitrate ion-supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol for the prevention or improvement of hypertension. <40> Non-therapeutic use of a combination of a nitrate ion-supplying 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 orally, 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 requiring it. <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 requiring them. <43> A method for improving the oral microbiome, 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 requiring such treatment. <44> A method for suppressing periodontal pathogen infection, comprising administering or ingesting, in an effective amount, a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to a subject requiring them. <45> A method for preventing or improving periodontal disease, comprising administering or ingesting, in an effective amount, a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to a subject in need. <46> A method for increasing nitric oxide, comprising administering or ingesting in an effective amount a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to a subject requiring it. <47> A method for preventing or improving 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. <48> A method for improving or enhancing motor function, comprising administering or ingesting, in an effective amount, a nitrate ion supplying compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol to a subject requiring such assistance. <49> <1> ~ <48> In this, the sugar alcohol is preferably erythritol, or erythritol and xylitol, or erythritol and mannitol, or erythritol, xylitol and mannitol. <50> <1> ~ <49> In this context, the nitrate ion supply compound is preferably a nitrate, and more preferably sodium nitrate or potassium nitrate. <51> <1> ~ <50> In this case, the ratio of the nitrate ion supply 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, and more preferably 20 parts by mass or less of the sugar alcohol per 1 part by mass of the nitrate ion supply compound. Also preferably 2.5 to 40 parts by mass, more preferably 5 to 20 parts by mass, and more preferably 10 to 20 parts by mass. <52> <1> ~ <50> In this study, the intake or administration of the nitrate ion supply 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, as the nitrate ion supply 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, as the sugar alcohol. <53> <3> , <11> , <19> , <27> or <35> In this context, improvement of the oral microbiota is defined as an increase in the content of oral Neisseria mucosa or Neisseria species in the oral cavity. [Examples]

[0036] The effects of combining nitrates and sugar alcohols on Neisseria mucosa in dental plaque biofilm microbiota. (1) Plaque collection and pretreatment Plaque was collected from the tooth surface of a healthy male in his 20s using a dental scaler, placed in a 1.5 mL tube, and washed twice by centrifugation at room temperature for 7,000 g with PBS(-) for 2 minutes. The pellet containing plaque bacteria was suspended in 300 μL of CELLBANKER 1Plus (Juji Field), and the prepared plaque suspension was stored at -80°C until use.

[0037] (2) Preparation of a plaque biofilm model by culturing plaque suspension Plaque suspensions were diluted 1000-fold in Brain Heart Infusion (BHI) medium (BD Japan) containing the following test compounds: potassium nitrate (Wako), erythritol (Wako), xylitol (Wako), mannitol (Wako), sorbitol (Wako), and maltitol (Wako), to prepare a culture medium containing plaque bacteria. One mL of the prepared culture medium was added to each of the Poly-L-Lysine Coat 24-Well Microplates (AGC TECHNO GLASS), and the plates were incubated at 37°C under microaerophilic conditions using an aneropack / bikouki (Mitsubishi Gas Chemical) for 48 hours to produce plaque biofilms. After 48 hours of incubation, the supernatant was carefully aspirated without disturbing the plaque biofilm at the bottom of the plate to prepare plaque biofilm samples.

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

[0039] (4) Determination of Neisseria mucosa and Neisseria species in genomic DNA samples The quantitative analysis of Neisseria mucosa and Neisseria species was performed using real-time PCR with a Taqman probe. For the specific detection of Neisseria mucosa, since only Neisseria mucosa possesses the nitrate reductase gene (narG) among Neisseria species (see Non-Patent Literature 1), a Taqman probe and primer were constructed targeting narG (Table 1). Additionally, a Taqman probe and primer were constructed to specifically detect the rRNA16S gene of all Neisseria species (Table 1). Furthermore, the Taqman probe and primer for detecting the rRNA16S gene of all bacteria were constructed using sequences previously reported (Nadkarni, MA, et al. Microbiology (2002) 148, 257-266) (Table 1). The reaction solution composition 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. The copy numbers of the Neisseria mucosa nitrate reductase gene (narG) and the rRNA16S gene of Neisseria species, as well as the copy number of the rRNA16S gene of all bacteria, were calculated, and the Neisseria mucosa content and Neisseria species content were calculated using the following formula.

[0043]

number

[0044] (5) Results Figures 1 and 2 show the effects of 1% (W / V) potassium nitrate and 5% (W / V) various sugar alcohols (erythritol, xylitol, mannitol, sorbitol, maltitol) on Neisseria mucosa and Neisseria species in the plaque biofilm flora. Figures 1 and 2 show that potassium nitrate, erythritol, xylitol, mannitol, sorbitol, and maltitol alone did not show a significant effect on the content of Neisseria mucosa and Neisseria species compared to the control group. Furthermore, the combined use of potassium nitrate with sorbitol and maltitol did not show a significant effect. However, the combined use of potassium nitrate with erythritol, xylitol, or mannitol resulted in a marked increase in the content of Neisseria mucosa and Neisseria species. Furthermore, Figures 3 and 4 show the dose-dependent effect of combining potassium nitrate and erythritol on increasing the content of Neisseria mucosa and Neisseria species bacteria. Figures 3 and 4 show that potassium nitrate alone showed only a weak increase in Neisseria mucosa and Neisseria species content at 0.2% potassium nitrate, but when used in combination with erythritol, a significant increase in Neisseria mucosa and Neisseria species content was observed in an erythritol dose-dependent manner.

Claims

1. An oral microbiome improving agent comprising a nitrate ion supply compound and one or more sugar alcohols selected from erythritol, xylitol, and mannitol as active ingredients, for the purpose of increasing nitric oxide, preventing or improving hypertension, or improving or enhancing motor function.

2. The oral microbiota improving agent according to Claim 1, wherein the improvement of the oral microbiota is an increase in Neisseria mucosa in the oral cavity.

3. The oral microbiota improving agent according to Claim 1, wherein the improvement of the oral microbiota is an increase in Neisseria species in the oral cavity.

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