Plaque formation inhibitors, sweetening compositions, food and beverages, supplements, toothpaste compositions, and mouthwash compositions

Abstract

To provide a plaque formation inhibitor that can suppress plaque formation to a high degree. [Solution] The plaque formation inhibitor contains at least one compound selected from cyclodextran and cyclodextran derivatives detected from a culture solution of Bacillus microorganisms using dextran as a substrate and / or a reaction solution of cyclic isomaltose synthase using dextran as a substrate, and at least one compound selected from isopropylmethylphenol, catechin, menthol, specific sugar alcohols (sugar alcohols with antibacterial effects), cetylpyridinium chloride, and berberine.

Description

[Technical Field]

[0001] The present invention relates to plaque formation inhibitors, sweetening compositions, food and beverages, supplements, toothpaste compositions, and mouthwash compositions. [Background technology]

[0002] Dental caries refers to tooth decay caused by biological factors, and is primarily known to be caused by demineralization of teeth due to acids produced by bacteria in the oral cavity from carbohydrates. Streptococcus mutans, the bacterium that causes dental caries, is a Gram-positive, facultative anaerobic streptococcus. This bacterium synthesizes glucan, a sticky polysaccharide, from sucrose using glucosyltransferase (GTF), which it produces itself. This glucan, along with other oral bacteria, forms a mass called plaque on the tooth surface. Within this plaque, acid is produced, which erodes the tooth surface, leading to demineralization and the formation of cavities. Inhibiting plaque is considered an effective means of preventing and improving such oral diseases.

[0003] It has been known for some cyclic oligosaccharides to inhibit the enzymatic activity of GTF in Streptococcus mutans and thereby suppress dental caries. For example, anticariogenic agents containing cyclic isomaltoligosaccharide as an active ingredient are known (see Patent Document 1). [Prior art documents] [Patent Documents]

[0004] [Patent Document 1] Patent No. 3400868 [Overview of the project] [Problems that the invention aims to solve]

[0005] In recent years, it has been pointed out that oral diseases such as dental caries may affect diseases other than oral diseases, and the medical importance of suppressing plaque and preventing dental caries is increasing.

[0006] Therefore, the present invention aims to provide a plaque formation inhibitor, etc., that can suppress plaque formation to a high degree. [Means for solving the problem]

[0007] As a result of diligent research, the inventors have discovered that by combining cyclodextran and / or cyclodextran derivatives with specific antimicrobial substances, a unique plaque formation inhibitory effect beyond the predictable range can be achieved, leading to the completion of the present invention. Specifically, the present invention provides the following:

[0008] The invention relating to the first feature comprises (A) at least one compound selected from cyclodextran and cyclodextran derivatives detected from the culture solution of Bacillus microorganisms using dextran as a substrate and / or the reaction solution of cyclic isomaltose synthase using dextran as a substrate, and (B) isopropylmethylphenol, triclosan, hinokitiol, thymol, 2-phenylphenol, chlorooxylenol, resorcinol, catechin, stilpenes, and cinnamic acid Benzoic acids, flavonoids, naphthoquinones, menthol, thymol, linatyl acetate, camphor, limonene, isomenthol, methyl salicylate, menthone, erythritol, xylitol, sorbitol, maltitol, lactitol, cetylpyridinium chloride, benzalkonium chloride, dodecylpyridinium chloride, chlorhexidine, dofemine bromide, tryptanthrin, berberine, plumbagin, lapachol, menadione, indole-3-carbinol and 3,The product contains at least one compound selected from 3'-diisodosolmethane, and if component (B) contains at least one selected from isopropylmethylphenol, triclosan, hinokitiol, thymol, 2-phenylphenol, chlorooxylenol and resorcinol, the concentration of component (A) is 1 ppm to 90 ppm per 100% of the plaque formation inhibitor, and the concentration of component (B) is 1 ppm to 90 ppm per 100% of the plaque formation inhibitor, and component (B) contains catechin, s If the product contains at least one selected from thyrylpenes, cinnamic acids, benzoic acids, flavonoids, and naphthoquinones, the concentration of component (A) is 5 ppm to 50 ppm relative to 100% of the plaque formation inhibitor, and the concentration of component (B) is 500 ppm to 2000 ppm relative to 100% of the plaque formation inhibitor, and if component (B) contains at least one selected from menthol, thymol, linatyl acetate, camphor, limonene, isomenthol, methyl salicylate, and menthone, the concentration of component (A) The concentration of component (B) is 1 ppm to 5000 ppm relative to 100% of the plaque formation inhibitor, and the concentration of component (B) is 800 ppm to 1200 ppm relative to 100% of the plaque formation inhibitor. If component (B) contains at least one selected from erythritol, xylitol, sorbitol, maltitol, and lactitol, the concentration of component (A) is 20 ppm to 500 ppm relative to 100% of the plaque formation inhibitor, and the concentration of component (B) is 9% to 15% relative to 100% of the plaque formation inhibitor. If component (B) contains at least one selected from cetylpyridinium chloride, benzalkonium chloride, dodecylpyridinium chloride, chlorhexidine, and dofemine bromide, the concentration of component (A) is 1 ppm to 5000 ppm per 100% of the plaque formation inhibitor, and the concentration of component (B) is 0.2 ppm to 20 ppm per 100% of the plaque formation inhibitor, and component (B) contains berberine, tryptanthrin, plumbagin, lapachol, menadione, indole-3-carbinol, and 3,When containing at least one selected from 3'-diisosordomethane, the concentration of the component (A) is 1 ppm or more and 100 ppm or less based on 100% of the dental plaque formation inhibitor, and the concentration of the component (B) is 0.1 ppm or more and 10 ppm or less based on 100% of the dental plaque formation inhibitor.,

[0009] The invention according to the second feature is an application example to a sweet composition, the invention according to the third feature is an application example to a food or drink, the invention according to the fourth feature is an application example to a supplement, the invention according to the fifth feature is an application example to a dentifrice composition, and the invention according to the sixth feature is an application example to a mouthwash composition.,

Effects of the Invention

[0010] According to the present invention, it is possible to provide a dental plaque formation inhibitor or the like that can suppress the formation of dental plaque at a high level.,

Brief Description of the Drawings

[0011] [Figure 1] FIG. 1 is an example of the structural formula of cyclodextrin (CI). [Figure 2] FIG. 2 is an example of the structural formula of cyclodextrin (CD). [Figure 3] FIG. 3 is an explanatory diagram showing criteria for determining the presence or absence of a specific effect (synergistic effect).

Modes for Carrying Out the Invention

[0012] Hereinafter, specific embodiments of the present invention will be described in detail with reference to the drawings. However, the present invention is not limited to the following embodiments, and can be implemented with appropriate modifications within the scope of the object of the present invention.,

[0013] <Dental Plaque Formation Inhibitor> The dental plaque formation inhibitor according to the present embodiment contains at least one compound selected from (A) cyclodextrin and / or its derivatives and (B) a specific antibacterial substance.,

[0014] [(A) Cyclodextran and / or its derivatives] Component (A) is at least one compound selected from cyclodextran and cyclodextran derivatives detected from the culture solution of Bacillus microorganisms using dextran as a substrate and / or the reaction solution of cyclic isomaltose synthase using dextran as a substrate. Figure 1 is an example of the structural formula of cyclodextran (CI). Cyclodextran is a cyclic isomaltoligosaccharide in which 4 to 33 glucose molecules are linked cyclically by α-1,6-glycosidic bonds.

[0015] Derivatives of cyclodextran include both naturally occurring compounds and artificially synthesized compounds. In cyclodextran derivatives, at least one of the alcoholic hydroxyl groups of cyclodextran is substituted with another functional group. The other functional group is not particularly limited, but may be one or more selected from chloro, bromo, iodo, alkoxy, glycosyl, thiol, sulfide, seleno, amino, amide, and phosphate groups. Among these, the functional group (substituent) substituted from the alcoholic hydroxyl group is preferably a polar group. These functional groups may substitute a single hydroxyl group of one molecule of cyclodextran, multiple hydroxyl groups may be substituted with the same functional group, or multiple hydroxyl groups may be substituted with different functional groups.

[0016] Preferred compounds as derivatives of cyclodextran include branched cyclodextran (where the alcoholic hydroxyl group is replaced by a glycosyl group) and amino-substituted cyclodextran (where the hydroxyl group of some glucose residues is replaced by an amino group). Among the monosaccharide residues included in the latter, those in which the hydroxyl group at position 2 of the glucose residue is replaced by an amino group are generally called glucosamine residues.

[0017] For example, cyclodextran and its derivatives can be obtained from the culture medium of Bacillus microorganisms that use dextran as a substrate, or from the reaction solution of cyclic isomaltoligosaccharide synthase that uses dextran as a substrate (see Japanese Patent Nos. 3075873 and 3117328).

[0018] Examples of Bacillus microorganisms capable of producing cyclodextran include strain Bacilluscirculans T-3040 (later renamed strain Paenibacillus agaridevorans T-3040) and strain Paenibacillus sp. 598K. An example of a cyclic isomaltoligosaccharide synthase is cyclic isomaltoligosaccharide glucanotransferase (CITase), obtained by purifying the culture medium of Bacillus microorganisms capable of producing cyclodextran.

[0019] As mentioned above, specific examples of preferred compounds as derivatives of cyclodextran include branched cyclodextran (where alcoholic hydroxyl groups are replaced with glycosyl groups) and amino-substituted cyclodextran (where hydroxyl groups of some glucose residues are replaced with amino groups). However, these are merely examples of known cyclodextran derivatives, and not all cyclodextran derivatives obtainable by microbial and / or enzymatic reactions have been identified.

[0020] Methods for measuring the state of cyclodextran derivatives include chromatography, mass spectrometry, and nuclear magnetic resonance (NMR) spectroscopy. However, these methods can only observe specific components, making it impossible to analyze all of the extremely large number of trace components, such as products resulting from microbial and / or enzymatic reactions. In particular, for carbohydrates like cyclodextran, the UV absorption in chromatography is weak, requiring the use of low-sensitivity IR detectors, making the detection of trace components itself difficult. Even if the main component can be detected, chromatography is not a method for identifying its structure, and there are currently no suitable columns available to separate the countless cyclodextran derivatives.

[0021] Furthermore, even when these methods are combined with mass spectrometry (LC-MS) for analysis, it is extremely difficult to clearly identify which parent peak each mass peak corresponds to based on data obtained from peaks with insufficient separation. In addition, cyclodextran derivatives are difficult to ionize, making the detection of trace components itself challenging. Moreover, while methods such as NMR are effective for structural analysis of carbohydrates, it is not practical to analyze all derivatives in a mixture of countless carbohydrates with unknown structures.

[0022] Even if it were possible to identify all cyclodextran derivatives produced by microbial and / or enzymatic reactions using equipment with extremely low detection limits, this would require an enormous number of trials and enormous costs, making it impractical.

[0023] Therefore, there are currently no appropriate means of measurement and analysis to comprehensively cover all types of cyclodextran derivatives contained in the reaction products of microbial and / or enzymatic reactions. Consequently, it is not possible to comprehensively describe the specific forms of cyclodextran derivatives contained in the reaction products of microbial and / or enzymatic reactions, and it is impossible or impractical to directly identify such substances by their structure or properties. It cannot be said that it is unclear to describe cyclodextran derivatives as "at least one compound detected from the culture solution of Bacillus microorganisms that use dextran as a substrate and / or the reaction solution of cyclic isomaltose synthase that uses dextran as a substrate."

[0024] (Note that the contents of the references are incorporated as part of this specification by reference, but matters specifically stated herein shall be governed by the provisions of this specification.) When cyclodextran is obtained using microorganisms or enzymes, it is obtained as a mixture of cyclodextran and a branched cyclodextran. This mixture of cyclodextran and a branched cyclodextran can be separated, if necessary, into a composition in which the concentration of cyclodextran is 75% or more, preferably 85% or more, more preferably 99% or more, or a composition in which the concentration of the branched cyclodextran is 75% or more, preferably 85% or more, more preferably 99% or more, by using conventionally known affinity chromatography or gel filtration chromatography. The plaque formation inhibitor according to this embodiment may be any of the following, as long as it does not impair the objective of the present invention: a mixture of cyclodextran and a branched cyclodextran; a composition in which the concentration of cyclodextran is 75% or more, preferably 85% or more, and more preferably 99% or more; or a composition in which the concentration of the branched cyclodextran is 75% or more, preferably 85% or more, and more preferably 99% or more.

[0025] Furthermore, in this specification, when "cyclodextran" is referred to, it refers to cyclodextran, which is a cyclic isomaltoligosaccharide that does not have a branched structure, and those that have a branched structure are distinguished from "cyclodextran" as "branched cyclodextran."

[0026] Cyclodextrin (CD) is known as a compound that is mutually similar to cyclodextran (CI). Figure 2 shows an example of the structural formula of cyclodextrin (CD).

[0027] CI and CD are both cyclic oligosaccharides in which multiple glucose molecules are linked in a ring, but they differ in the following ways: Firstly, CI consists of 4 to 33 glucose molecules, while CD consists of 6 to 8 glucose molecules. Secondly, in CI, the glucose molecules are linked by α-1,6-glycosidic bonds, while in CD, they are linked by α-1,4-glycosidic bonds. Thirdly, the molecular structure of CI has a large and shallow opening, while the molecular structure of CD has a small and deep opening. Fourthly, CI is highly soluble in water, while CD is poorly soluble in water.

[0028] [(B) Specific antimicrobial substances] The plaque formation inhibitor contains at least one compound selected from isopropylmethylphenol, catechin, menthol, certain sugar alcohols, cetylpyridinium chloride, and berberine.

[0029] The mechanism of antibacterial action can be broken down into the following eight items. (1) It disrupts the integrity of the lipid layer of the cell membrane and increases membrane permeability, thus hindering the exchange of substances inside and outside the cell. (2) Disrupting the cell membrane causes intracellular substances to leak out. (3) Suppresses spore growth. (4) The pathogen is initially suppressed by attracting an electric charge to the electronegative components on the bacterial surface, and once a sufficient concentration is reached, the charge is neutralized, causing the membrane components to lose their order and organization. (5) Activates latent ribonucleases in the cells of microorganisms, thereby promoting cellular autolysis. (6) It causes intercalation via binding to DNA, thereby inhibiting nucleic acid synthesis. (7) It inhibits cell membrane permeability and induces the accumulation of reactive oxygen species (ROS), thereby causing apoptosis in bacteria. (8) Inhibiting the activity of COX-2, a type of cyclooxygenase, suppresses the production of prostaglandin E2 (PGE2), creating an unfavorable environment for bacterial infection.

[0030] The inventors investigated representative compounds exhibiting the action of each mechanism and discovered a new finding: the combined use of cyclodextran and / or cyclodextran derivatives with specific antimicrobial agents comprehensively covers antimicrobial activity. These antimicrobial agents are described below.

[0031] [Isopropylmethylphenol, etc.] Isopropylmethylphenol (C 10 H 14 O,Isopropyl Methylphenol (IPMP) is a phenolic compound with a structure in which an isopropyl group and a methyl group are substituted for phenol.

[0032] IPMPs directly attack the cell membrane, increasing its permeability and thereby hindering the exchange of substances inside and outside the cell. This antimicrobial mechanism is known to be due to hydrophobic interactions with the lipid layer of the cell membrane. (H. Ohara et al., Antimicrobial effect of oral care gel containing hinokitiol and 4-isopropyl-3-methylphenol against intraoral pathogenic microorganisms. PLOS ONE 2023:Sep.1, https: / / journals.plos.org / plosone / article?id=10.1371 / journal.pone.0283295)

[0033] Compounds structurally similar to IPMP include triclosan, hinokitiol, thymol, 2-phenylphenol, chlorooxylenol, and resorcinol.

[0034] When component (A) is used in combination with IPMP, etc., the concentration of component (A) is preferably 1 ppm or more, more preferably 10 ppm or more, and even more preferably 40 ppm or more relative to the total. On the other hand, the concentration of component (A) is preferably 90 ppm or less, and more preferably 70 ppm or less, relative to the total.

[0035] (B) The concentration of component IPMP, etc., is preferably 1 ppm or more, more preferably 10 ppm or more, and even more preferably 30 ppm or more, relative to the total. On the other hand, the concentration of IPMP, etc., is preferably 90 ppm or less, and more preferably 70 ppm or less.

[0036] According to the Ministry of Health, Labour and Welfare's standards for approval of the manufacture and sale of medicated toothpastes, the antimicrobial substances used in toothpastes are standardized in the IPMP range of 20 to 1000 ppm or less. These antimicrobial agents, with their broad antimicrobial spectrum, may have a non-specific bactericidal effect not only against the target caries-causing bacteria but also against beneficial bacteria in the oral cavity, such as Veillonella and Neisseria, at concentrations within this range. Bacteria such as Veillonella and Neisseria have been shown to reduce lactic acid levels in the oral microbiome and are expected to contribute to caries prevention by metabolizing nitrates derived from vegetables and saliva into antimicrobial nitrites. Therefore, it is not considered advisable to kill these bacteria as well.

[0037] From this perspective, it is even more preferable that the concentration of IPMP, etc., be 20 ppm or less. This gives the plaque formation inhibitor described herein a significant advantage in that it does not kill beneficial bacteria in the oral cavity and can synergistically inhibit the formation of plaque (biofilm).

[0038] Here, "whole" refers to the entire dosage form, and the same applies hereafter. If the dosage form is a plaque formation inhibitor, "whole" refers to 100% of the plaque formation inhibitor; if the dosage form is a food or beverage, "whole" refers to 100% of the food or beverage; if the dosage form is a supplement, "whole" refers to 100% of the supplement; if the dosage form is a toothpaste composition, "whole" refers to 100% of the toothpaste composition; and if the dosage form is a mouthwash composition, "whole" refers to 100% of the mouthwash composition.

[0039] [Catechins, etc.] Catechin (C 15 H 14O6) is a type of plant-derived polyphenol and is a flavanol (a type of flavonoid) that has a benzene ring. Catechin has several isomers, including catechin (C), epicatechin (EC), gallocatechin (GC), epigallocatechin (EGC), epicatechin gallate (ECG), epigallocatechin gallate (EGCG), epigallocatechin gallate (EGCG), and EGCG (epigallocatechin gallate). In this specification, catechin refers not only to catechin (C) in the narrow sense, but also to all catechins in the broad sense, including multiple isomers.

[0040] The antibacterial mechanism of catechins is due to the filtration of intracellular substances by disrupting the cell membrane (from the Japan Catechin Society website, Top page > Effects and Actions of Catechins > Antibacterial and Bactericidal Effects, https: / / www.catechin-society.com / effect_01.html).

[0041] Many polyphenols have similar structural properties, and many of them have been reported to have antibacterial effects. Examples include stilpenes containing resveratrol, cinnamic acid derivatives containing cinnamic acid, benzoic acid derivatives containing gallic acid, flavonoids containing catechins, and naphthoquinones containing naphthalene. (LBChibane et al., Antibacterial Properties of Polyphenols: Characterization and QSAR (Quantitative Structure-Activity Relationship) Models. Frontiers in Microbiology. 2019; Vol.10.)

[0042] When component (A) is used in combination with catechin, etc., the concentration of component (A) is preferably 5 ppm or more relative to the total. On the other hand, the concentration of component (A) is preferably 50 ppm or less relative to the total, more preferably 30 ppm or less, and even more preferably 10 ppm or less.

[0043] (B) The concentration of catechin and the like, which is component (B), is preferably 500 ppm or more of the total, more preferably 700 ppm or more, and even more preferably 750 ppm or more. On the other hand, the concentration of catechin and the like, which is preferably 2000 ppm or less of the total, more preferably 1500 ppm or less, and even more preferably 1000 ppm or less.

[0044] [Menthol, etc.] Menthol (C 10 H 20 L-menthol (L-Menthol) is a terpenoid with a structure in which a hydroxyl group (-OH) is bonded to a cyclohexane ring, and is a naturally occurring organic compound found in mint. While L-menthol (natural form) is primarily used, synthetic menthol also exists. In this specification, menthol includes not only natural L-menthol but also synthetic menthol.

[0045] It is hypothesized that the antibacterial effect of menthol arises, at least in part, from the disruption of the lipid fraction of microbial cell membranes, resulting in altered membrane permeability and leakage of intracellular substances. (D. Trombetta et al., Mechanisms of Antibacterial Action of Three Monoterpenes. Antimicrobial Agents and Chemotherapy. 2005;49;6)

[0046] Substances with a structure similar to menthol include thymol, linatyl acetate, camphor, limonene, isomenthol, methyl salicylate, and menthone.

[0047] When using the component (A) in combination with menthol or the like, the concentration of the component (A) is preferably 1 ppm or more, more preferably 10 ppm or more, still more preferably 200 ppm or more, even more preferably 500 ppm or more, and particularly preferably 1000 ppm or more with respect to the whole. On the other hand, the concentration of the component (A) is preferably 5000 ppm or less, more preferably 3000 ppm or less with respect to the whole.

[0048] The concentration of menthol or the like as the component (B) is preferably 800 ppm or more, more preferably 900 ppm or more with respect to the whole. On the other hand, the concentration of menthol or the like is preferably 1200 ppm or less, more preferably 1100 ppm or less with respect to the whole.

[0049] [Specific sugar alcohol (sugar alcohol having antibacterial effect)] As the specific sugar alcohol, erythritol (C4H 10 O4, Erythritol), xylitol (C5H 12 O5, Xylitol), sorbitol (C6H 14 O6, Sorbitol), maltitol (C 12 H 24 O 11 , Maltitol) and lactitol (C 12 H 24 O 11 , Lactitol) can be mentioned.

[0050] Regarding the antibacterial action of the sugar alcohol, for example, xylitol effectively inhibits the growth of spores, and bacteriostatic activity against thermophilic spore-forming bacteria has been reported. (J. Piao et al., Bacteriostatic Activities of Monoacyl Sugar Alcohols against Thermophilic Sporeformers. Bioscience, Biotechnology, and Biochemistry. 2006; 70.1: 263 - 265)

[0051] When component (A) and sugar alcohols are used in combination, the concentration of component (A) is preferably 20 ppm or more, and more preferably 50 ppm or more, relative to the total. On the other hand, the concentration of component (A) is preferably 500 ppm or less, and more preferably 300 ppm or less, relative to the total.

[0052] (B) The concentration of the sugar alcohol, which is component (B), is preferably 9% or more of the total. On the other hand, the concentration of the sugar alcohol is preferably 15% or less of the total, and more preferably 12% or less.

[0053] [Cetylpyridinium chloride, etc.] Cetylpyridinium chloride (C 21 H 38 ClN,Cetylpyridinium Chloride (CPC) is a quaternary ammonium salt in which a pyridine ring and a cetyl group are bonded, and a chloride ion is associated with it. It is known as a cationic surfactant.

[0054] The following has been reported in previous literature regarding the antibacterial activity of CPC: Application of 0.8% CPC leads to the loss of electrostatic repulsion between individual cells, the loss of integrity of the microbial membrane, and ultimately, emulsification of membrane lipids along with leakage of cytoplasmic contents. It has been shown that CPC inhibits the pathogen by initially attracting charge to the electronegative components on the surface of Salmonella bacteria, and at sufficient concentrations, the neutralization of the charge leads to the loss of order and organization of membrane components. (Y. Yegin. et al., Cetylpyridinium chloride produces increased zeta-potential on Salmonella Typhimurium cells, a mechanism of the pathogen's inactivation. npj Science of Food. 2019;3:21)

[0055] Furthermore, at low concentrations, CPC activates latent ribonucleases within microorganisms, promoting cellular autodegradation. At high concentrations, CPC is known to exhibit antibacterial activity by forming vesicle-like structures on the bacterial cell surface, leading to leakage of cytoplasmic contents. (Nasila K. et al., A Review on Cetylpyridinium Chloride. International Journal of Research and Review. 2021:April.vol8.4)

[0056] Compounds with a structure similar to CPC include benzalkonium chloride, dodecylpyridinium chloride, chlorhexidine, and dofemine bromide.

[0057] When component (A) is used in combination with CPC, etc., the concentration of component (A) is preferably 1 ppm or more relative to the total, and more preferably 0.2 ppm or more. On the other hand, the concentration of component (A) is preferably 5000 ppm or less relative to the total, more preferably 2000 ppm or less, and even more preferably 500 ppm or less.

[0058] (B) The concentration of component CPC, etc., is preferably 0.2 ppm or higher relative to the total. On the other hand, the concentration of CPC, etc., is preferably 20 ppm relative to the total.

[0059] According to the Ministry of Health, Labour and Welfare's standards for approval of the manufacture and sale of medicated toothpastes, antimicrobial substances used in toothpastes are standardized in the CPC range of 10 to 50 ppm or less. These antimicrobial agents, with their broad antimicrobial spectrum, may have a non-specific bactericidal effect not only against the target caries-causing bacteria but also against beneficial bacteria in the oral cavity, such as Veillonella and Neisseria, at concentrations within this range. Bacteria such as Veillonella and Neisseria have been shown to reduce lactic acid levels in the oral microbiome and are expected to contribute to caries prevention by metabolizing nitrates derived from vegetables and saliva into antimicrobial nitrites. Therefore, it is not considered advisable to kill these bacteria as well.

[0060] From this perspective, it is more preferable that the concentration of CPC, etc., be 10 ppm or less. This gives the plaque formation inhibitor described herein a significant advantage in that it does not kill beneficial bacteria in the oral cavity and can synergistically inhibit the formation of plaque (biofilm).

[0061] [Berberine, etc.] Berberine (C 20 H 18 NO4,berberine is a type of benzylisoquinoline alkaloid and is a compound found in plants such as Phellodendron amurense (Rutaceae) and Coptis japonica (Ranunculaceae).

[0062] Berberine derivatives undergo DNA interaction and exhibit inhibitory activity against bacteria and fungi. (GBZhang et al., Discovery of natural berberine-derived nitroimidazoles as potentially multi-targeting agents against drug-resistant Escherichia coli. Scinece China Chemistriy.2018:61,557-568)

[0063] Furthermore, berberine and its derivatives are known to interact with bacterial DNA and promote the production of reactive oxygen species (ROS), thereby inducing cell death. (GFDu.et al., Proteomic investigation into the action mechanism of berberine against Streptococcus pyogenes. Journal of Proteomics. 2020;215:20)

[0064] Furthermore, berberine and berberine derivatives inhibit the activity of cyclooxygenase-2 (COX-2), which is an inflammatory mediator involved in the production of prostaglandins (particularly PGE2) within cells. PGE2 plays a role in promoting host responses and the inflammatory environment in bacterial infections. Excessive production of PGE2 during bacterial infection exacerbates inflammation and promotes bacterial growth. Berberine and berberine derivatives are known to suppress PGE2 production by inhibiting COX-2, and are therefore expected to exert antibacterial effects. (X. Liu et al., Berberine Inhibits Invasion and Metastasis of Colorectal Cancer Cells via COX-2 / PGE2 Mediated JAK2 / STAT3 Signaling Pathway. PLOS one. 2015; May 8)

[0065] Compounds with a structure similar to berberine include tryptanthrin, plumbagin, lapachol, menadione, indole-3-carbinol, and 3,3'-diisodosolmethane.

[0066] When using component (A) in combination with berberine, etc., the concentration of component (A) is preferably 1 ppm or more relative to the total.

[0067] (B) The concentration of component berberine, etc., is preferably 0.1 ppm or more relative to the total. On the other hand, the concentration of berberine, etc., is preferably 10 ppm or less relative to the total, and more preferably 3 ppm or less.

[0068] In this invention, the plaque formation inhibitor contains (B) a specific antimicrobial substance, which suppresses the growth of caries-causing bacteria, including Streptococcus mutans, and reduces the expression of glucosyltransferase (GTF). As a result, it is believed that (A) cyclodextran and / or its derivatives can exert a high plaque formation inhibitory effect with a smaller amount compared to when used alone.

[0069] Furthermore, it is thought that component (A) inhibits GTF and suppresses the amount of biofilm formation, thereby weakening the resistance of caries-causing bacteria to antimicrobial substances, allowing component (B) to exert a greater effect.

[0070] It is presumed that one or both of these hypotheses enabled the unique plaque formation inhibitory effect, exceeding the predictable range, compared to using either component (A) or component (B) alone.

[0071] <Uses of plaque formation inhibitors> Plaque formation inhibitors can be used in applications such as sweetening compositions, foods and beverages, supplements, toothpaste compositions, and mouthwash compositions.

[0072] [Sweet composition] The sweetening composition contains the plaque formation inhibitor described above and optionally (C) carbohydrates.

[0073] [Carbohydrates] The carbohydrate is not particularly limited, as long as it is different from the carbohydrates contained in the plaque formation inhibitors mentioned above, specifically, different from cyclodextran and its derivatives, as well as erythritol, xylitol, sorbitol, maltitol, and lactitol.

[0074] Carbohydrates include sugars, oligosaccharides, polysaccharides, sugar alcohols, and others. Sugars are a general term for substances remaining after removing oligosaccharides, polysaccharides, sugar alcohols, and others from carbohydrates, and include monosaccharides and disaccharides.

[0075] [Sugars] Sugars include monosaccharides and disaccharides.

[0076] (Monosaccharides) Monosaccharides are broadly classified into aldoses and ketoses. Aldoses are C n H 2n O n Aldoses are monosaccharides represented by (where n is an integer greater than or equal to 3) and have one aldehyde group at their terminus. More specifically, examples of monosaccharides that are aldoses include glyceraldehyde (C3), erythrose (C4), threose (C4), ribose (C5), arabinose (C5), xylose (C5), lyxose (C5), allose (C5), altrose (C6), glucose (C6), mannose (C6), glucose (C6), idose (C6), galactose (C6), talose (C6), etc.

[0077] Regarding ketoses, C is the same as for aldoses. n H 2n O n Although represented as (where n is an integer of 3 or greater), it refers to a monosaccharide that has one keto group (ketonic carbonyl group) inside its chain structure. More specifically, examples of monosaccharides that are ketoses include dihydroxyacetone (C3), erythrulose (C4), xylulose (C5), ribulose (C5), psicose (C6), fructose (C6), sorbose (C6), tagatose (C6), etc.

[0078] (disaccharide) Disaccharides have monosaccharides that are aldoses and / or ketoses as constituent sugars. Specifically, examples include sucrose, lactose, maltose, trehalose, cellobiose, isomaltose, partinose, xylobiose, laminaribiose, genthiobiose, turanose, maltulose, and palatinose.

[0079] [Oligosaccharides] Oligosaccharides that can produce a mixture containing aldoses and / or ketoses by hydrolysis refer to oligosaccharides having monosaccharides that are aldoses and / or ketoses as constituent sugars. More specifically, examples include cellobiose, trehalose, isomaltose, partinose, xylobiose, laminalibiose, genthiobiose, turanose, maltulose, palatinose, galactooligosaccharides, fructooligosaccharides, lactosucrose, isomaltoligosaccharides, and kestose. Cyclodextrins, such as cyclodextrins, are also included.

[0080] [Cyclic oligosaccharides] Cyclic oligosaccharides that can produce a mixture containing aldoses and / or ketoses upon hydrolysis refer to cyclic oligosaccharides having monosaccharides, which are aldoses and / or ketoses, as constituent sugars. Cyclodextrins, as mentioned above, are typical examples of cyclic oligosaccharides, and among these, those with 6, 7, or 8 glucose units linked are known. These cyclodextrins are sometimes referred to as α-cyclodextrin (cyclohexamylose, α-CD; with 6 glucose units linked), β-cyclodextrin (cycloheptamylose, β-CD; with 7 glucose units linked), and γ-cyclodextrin (cyclooctaamylose, γ-CD; with 8 glucose units linked), depending on the context. However, cyclic oligosaccharides are not limited to cyclodextrins; they may also be cyclic oligosaccharides having monosaccharides other than glucose as constituent sugars.

[0081] [Polysaccharide] Examples of polysaccharides include reduced maltose syrup, sorbitol, lactitol, xylitol, and maltitol.

[0082] [Polysaccharide] Polysaccharides that can produce a mixture containing aldoses and / or ketoses by hydrolysis refer to polysaccharides that have monosaccharides consisting of aldoses and / or ketoses as constituent sugars. More specifically, examples include amylose, amylopectin, glycogen, and galactogen. Of these, amylose, amylopectin, and glycogen are all polysaccharides that have glucose as a constituent sugar, and tend to differ from each other in terms of branching structure, etc. Galactogen, on the other hand, is a polysaccharide that has galactose as a constituent sugar.

[0083] [Sugar alcohols] Examples of sugar alcohols include reduced maltose syrup.

[0084] [others] In addition to the monosaccharides, disaccharides, oligosaccharides, cyclic oligosaccharides, and polysaccharides mentioned above, aspartame, acesulfame potassium, stevia, and others are also included in the category of carbohydrates.

[0085] The carbohydrate content is not particularly limited, as long as it does not affect its function as a plaque formation inhibitor.

[0086] [Food and beverages] Beverages containing plaque formation inhibitors are not particularly limited and include soft drinks, vegetable / fruit juices, alcoholic beverages, dairy beverages, lactic acid bacteria beverages, teas, coffee beverages, etc. Additives commonly found in beverages, such as antioxidants, flavorings, inorganic acids, inorganic acid salts, inorganic salts, colorants, emulsifiers, preservatives, seasonings, sweeteners, acidulants, fruit extracts, vegetable extracts, nectar extracts, lactic acid bacteria, pH adjusters, and quality stabilizers, may be added individually or in combination.

[0087] Furthermore, foods containing plaque formation inhibitors are not particularly limited and include not only finished products but also semi-finished products. Examples include frozen foods, frozen dough, canned foods, bottled foods, chilled foods, pressure-heated sterilized foods, dried foods, processed meat products, processed seafood products, processed vegetable products, processed fruit products, processed grain products, seasonings, beverages, baked goods, Japanese sweets, Western sweets, chocolate, semi-chocolate, chocolate confectionery, semi-chocolate confectionery, edible film, candies (gummies, hard candies, etc.), gum, tablets, jelly, yogurt, ice cream, ice milk, lacto ice, frozen desserts, cream, butter, butter oil, cheese, concentrated whey, concentrated milk, skimmed concentrated milk, unsweetened condensed milk, unsweetened skimmed condensed milk, sweetened condensed milk, sweetened skimmed condensed milk, whole milk powder, skimmed milk powder, premixed products, cream powder, whey powder, protein-concentrated whey powder, buttermilk powder, sweetened milk powder, prepared milk powder, prepared liquid milk, fermented milk, lactic acid bacteria beverages, pet food, etc.

[0088] 〔supplement〕 Furthermore, examples of supplements containing plaque formation inhibitors include base supplements, health supplements, and optional supplements, and examples of nutritional functional food components contained in each include vitamins, minerals, amino acids, enzymes, dietary fiber, coenzyme Q10, placenta, hyaluronic acid, collagen, collagen peptides, astaxanthin, omega-3 fatty acids, glucosamine, and chondroitin, as long as they do not impair the effects of the present invention.

[0089] [Other ingredients that may be included in sweetening compositions, foods, and supplements] Sweetening compositions, foods, and supplements containing plaque formation inhibitors can be classified as food additives, foods, and nutritional supplements, respectively, and may contain additives other than components (A) and (B) that are commonly used in the fields of food additives, foods, and nutritional supplements. Such additives are not particularly limited as long as they do not impair the purpose of the present invention, but examples include acidulants, enzymes, spices, pigments, preservatives, flavorings, sweeteners, seasonings, emulsifiers, thickeners, stabilizers, foaming agents, leavening agents, colorants, oxidizing agents, preservatives, confectionery gelatin, confectionery albumin, confectionery starch, sterilizers, antioxidants, and fungicides. Furthermore, sweetening compositions, foods, and supplements containing plaque formation inhibitors according to this embodiment may contain linear, branched, or cyclic polysaccharides that are structurally similar to component (A), to the extent that they do not impair the purpose of the present invention.

[0090] [Toothpaste composition, mouthwash composition] The plaque formation inhibitor according to this embodiment may be in the form of a toothpaste composition or a mouthwash composition. These toothpaste compositions and mouthwash compositions may be classified as pharmaceuticals, quasi-drugs, or cosmetics, and may contain medicinal ingredients or cosmetic ingredients.

[0091] The toothpaste composition and mouthwash composition according to this embodiment may, if necessary, be used in combination with other medicinal ingredients to treat, suppress, or prevent oral diseases such as bad breath, periodontal disease, and hypersensitivity, or to strengthen tooth enamel. Examples of medicinal ingredients that may be contained in the toothpaste composition and mouthwash composition according to this embodiment include conventionally used ones such as fluoride, xylitol, propolis, Swertia japonica extract, and hydroxyapatite. Specific embodiments of the toothpaste composition and mouthwash composition include oral care products such as toothpaste, mouthwash, gargle, and mouth spray, as well as oral care foods such as edible films, tablets, chewables, candies, gummies, gum, and beverages.

[0092] The toothpaste composition and mouthwash composition according to this embodiment may contain any conventionally used additives, depending on their form. Such additives are not limited as long as they do not hinder the objective of the present invention, but may include abrasives, binders, colorants, fragrances, sweeteners, etc. Furthermore, the caries inhibitor, toothpaste composition, and mouthwash composition according to this embodiment may contain linear, branched, or cyclic polysaccharides that are structurally similar to component (A), to the extent that they do not hinder the objective of the present invention. [Examples]

[0093] The present invention will be specifically described below with reference to examples, but the present invention is not limited to these examples.

[0094] <Test 1> CI-Dextran mix + Isopropylmethylphenol (IPMP) [Table 1] [Table 2]

[0095] As shown in Table 1, samples were prepared using four types of CI-Dextran mix (manufactured by Wellneo Sugar Co., Ltd., abbreviated as CI mix as appropriate, with a CI concentration of approximately 13%): 0.01%, 0.025%, 0.05%, and 0.075% (of which the net CI concentrations were 13 ppm, 32.5 ppm, 65 ppm, and 97.5 ppm). Samples were also prepared using two types of IPMP: 50 ppm and 100 ppm. As shown in Table 2, for each concentration, four types of samples were prepared with and without the addition of CI mix and with and without the addition of IPMP.

[0096] For example, for a CI mix concentration of 0.010% and an IPMP concentration of 50 ppm, samples were prepared in four types: A) CI mix 0%, IPMP 0 ppm, B) CI mix 0%, IPMP 50 ppm, C) CI mix 0.010%, IPMP 0 ppm, and D) CI mix 0.010%, IPMP 50 ppm.

[0097] For each sample, Streptococcus mutans was inoculated onto THB medium (Trypticase Soy Broth medium, manufactured by Becton Dickinson, Inc.) and pre-cultured under anaerobic conditions for 16 hours. After that, the bacterial count of S. mutans was measured using 1 / 4 THB medium containing 0.5% sucrose until it reached 1.0 × 10⁶. 7 A suspension was prepared to a concentration of 100 cells / mL, each sample was mixed, and seeded in a 96-well plate. The plate was incubated for 24 hours at 37°C under anaerobic conditions. After discarding the culture supernatant and washing with distilled water, a 0.1% crystal violet solution was added to the biofilm remaining on the bottom of the plate and allowed to stand for 15 minutes to fix the dye. The dye solution was then discarded and washed with distilled water, and the plate was immersed in ethanol for 15 minutes to elute the dye. The amount of dye was measured by absorbance (590 nm).

[0098] As shown in Figure 3, sample A' (with IPMP added, without CI mix) should suppress biofilm production more effectively than sample A (without IPMP added, without CI mix), resulting in lower turbidity in the post-culture solution. Let a be the magnitude (slope) of the decrease in turbidity in the post-culture solution. Similarly, sample B' (with IPMP added, with CI mix added) should suppress biofilm production more effectively than sample B (without IPMP added, with CI mix added), resulting in lower turbidity in the post-culture solution. Let b be the magnitude (slope) of the decrease in turbidity in the post-culture solution.

[0099] If a ≥ b (b / a ≤ 1), then it cannot be said that there is a specific effect (synergistic effect) that exceeds expectations, and a<b(b / a> 1) If this is true, then a specific effect can be said to exist. Therefore, the presence or absence of a specific effect was evaluated based on the magnitude of b / a. The results are shown in Table 1.

[0100] As shown in Table 1, it was confirmed that when the concentration of component (A) is between 1 ppm and 90 ppm relative to the total, and the concentration of component (B), IPMP, is between 1 ppm and 90 ppm relative to the total, a specific plaque formation inhibitory effect can be exerted that exceeds the effect of combinations predictable from component (A) alone or component (B) alone. In particular, it was confirmed that a more remarkable effect can be exerted when the concentration of component (A) is between 40 ppm and 70 ppm relative to the total, and the concentration of component (B), IPMP, is between 30 ppm and 70 ppm relative to the total.

[0101] <Test 2> CI-Dextran mix + catechin [Table 3]

[0102] The b / a values ​​were evaluated using the same method as in Experiment 1, except that IPMP was replaced with catechin and the concentrations of CI-Dextran mix and catechin were as shown in Table 3. The results are shown in Table 3.

[0103] As shown in Table 3, it was confirmed that when the concentration of component (A) is between 5 ppm and 50 ppm relative to the total, and the concentration of component (B), catechin, is between 500 ppm and 2000 ppm relative to the total, a specific plaque formation inhibitory effect exceeding the effects of combinations predictable from component (A) alone or component (B) alone can be achieved. In particular, it was confirmed that a more remarkable effect can be achieved when the concentration of component (A) is between 5 ppm and 10 ppm relative to the total, and the concentration of component (B), catechin, is between 750 ppm and 1000 ppm relative to the total.

[0104] <Test 3> CI-Dextran mix + menthol [Table 4]

[0105] The b / a values ​​were evaluated using the same method as in Experiment 1, except that IPMP was replaced with menthol and the concentrations of CI-Dextran mix and menthol were as shown in Table 4. The results are shown in Table 4.

[0106] As shown in Table 4, it was confirmed that when the concentration of component (A) is between 1 ppm and 5000 ppm relative to the total, and the concentration of component (B), menthol, is between 800 ppm and 1200 ppm relative to the total, a specific plaque formation inhibitory effect exceeding the effects predictable from component (A) alone or combinations of component (B) alone can be achieved. In particular, it was confirmed that a more remarkable effect was achieved when the concentration of component (A) is between 500 ppm and 5000 ppm relative to the total, and the concentration of component (B), menthol, is between 800 ppm and 1200 ppm relative to the total, and an even more remarkable effect was achieved when the concentration of component (A) is between 1000 ppm and 5000 ppm relative to the total, and the concentration of component (B), IPMP, is between 800 ppm and 1200 ppm relative to the total.

[0107] <Test 4> CI-Dextran mix + erythritol [Table 5]

[0108] The b / a values ​​were evaluated using the same method as in Experiment 1, except that IPMP was replaced with erythritol (ERT) and the concentrations of CI-Dextran mix and erythritol were as shown in Table 5. For ERT, "Erythritol SS" (Tsuruya Chemical Industries Co., Ltd.) was used. The results are shown in Table 5.

[0109] As shown in Table 5, it was confirmed that when the concentration of component (A) is between 20 ppm and 500 ppm relative to the total, and the concentration of component (B), ERT, is between 9% and 15% relative to the total, a specific plaque formation inhibitory effect exceeding the effects of component (A) alone or combinations of component (B) alone can be achieved. In particular, it was confirmed that a more remarkable effect can be achieved when the concentration of component (A) is between 50 ppm and 500 ppm relative to the total, and the concentration of component (B), ERT, is between 9% and 12% relative to the total.

[0110] <Test 5> CI-Dextran mix + CPC [Table 6]

[0111] The b / a values ​​were evaluated using the same method as in Experiment 1, except that IPMP was replaced with CPC and the concentrations of CI-Dextran mix and CPC were as shown in Table 6. The results are shown in Table 6.

[0112] As shown in Table 6, it was confirmed that when the concentration of component (A) is between 1 ppm and 5000 ppm relative to the total, and the concentration of component (B), CPC, is between 0.2 ppm and 20 ppm relative to the total, a specific plaque formation inhibitory effect exceeding the effects of combinations predictable from component (A) alone or component (B) alone can be achieved. In particular, it was confirmed that a more remarkable effect can be achieved when the concentration of component (A) is between 1 ppm and 2000 ppm relative to the total, and the concentration of component (B), CPC, is between 0.2 ppm and 20 ppm relative to the total.

[0113] <Study 6> CI-Dextran mix + tryptanthrin [Table 7]

[0114] The b / a values ​​were evaluated using the same method as in Experiment 1, except that IPMP was replaced with tryptanthrin and the concentrations of CI-Dextran mix and tryptanthrin were as shown in Table 7.

[0115] As shown in Table 7, it was confirmed that when the concentration of component (A) is between 1 ppm and 100 ppm relative to the total, and the concentration of component (B), tryptanthrin, is between 0.1 ppm and 10 ppm relative to the total, a specific plaque formation inhibitory effect exceeding the effects of combinations predictable from component (A) alone or component (B) alone can be achieved. In particular, it was confirmed that a more remarkable effect can be achieved when the concentration of component (A) is between 1 ppm and 70 ppm relative to the total, and the concentration of component (B), tryptanthrin, is between 1 ppm and 3 ppm relative to the total.

Claims

[Claim 1] (A) at least one compound selected from cyclodextran and cyclodextran derivatives detected from the culture solution of Bacillus microorganisms using dextran as a substrate and / or the reaction solution of cyclic isomaltose synthase using dextran as a substrate, (B) At least one compound selected from isopropylmethylphenol, triclosan, hinokitiol, thymol, 2-phenylphenol, chloroxylenol, resorcinol, menthol, thymol, linatyl acetate, camphor, limonene, isomenthol, methyl salicylate, menthone, erythritol, xylitol, sorbitol, maltitol, lactitol, cetylpyridinium chloride, benzalkonium chloride, dodecylpyridinium chloride, chlorhexidine, dofemine bromide, berberine, tryptanthrin, plumbagin, lapachol, menadione, indole-3-carbinol and 3,3'-diisodosolmethane, It contains, If component (B) contains at least one selected from isopropylmethylphenol, triclosan, hinokitiol, thymol, 2-phenylphenol, chlorooxylenol, and resorcinol, the concentration of component (A) is 13 ppm to 90 ppm relative to 100% of the plaque formation inhibitor, and the concentration of component (B) is 50 ppm to 90 ppm relative to 100% of the plaque formation inhibitor. If component (B) contains at least one selected from menthol, thymol, linatyl acetate, camphor, limonene, isomenthol, methyl salicylate, and menthone, the concentration of component (A) is 13 ppm to 1300 ppm relative to 100% of the plaque formation inhibitor, and the concentration of component (B) is 800 ppm to 1200 ppm relative to 100% of the plaque formation inhibitor. If component (B) contains at least one selected from erythritol, xylitol, sorbitol, maltitol, and lactitol, the concentration of component (A) is 20 ppm to 130 ppm relative to 100% of the plaque formation inhibitor, and the concentration of component (B) is 9% to 10% relative to 100% of the plaque formation inhibitor. If component (B) contains at least one selected from cetylpyridinium chloride, benzalkonium chloride, dodecylpyridinium chloride, chlorhexidine, and dofemine bromide, the concentration of component (A) is 13 ppm to 1300 ppm relative to 100% of the plaque formation inhibitor, and the concentration of component (B) is 0.2 ppm to 10 ppm relative to 100% of the plaque formation inhibitor. A plaque formation inhibitor in which, if component (B) contains at least one selected from berberine, tryptanthrin, plumbagin, lapachol, menadione, indole-3-carbinol, and 3,3'-diisodosolmethane, the concentration of component (A) is 1 ppm or more and 100 ppm or less per 100% of the plaque formation inhibitor, and the concentration of component (B) is 0.1 ppm or more and 5 ppm or less per 100% of the plaque formation inhibitor.

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