Composition for controlling bacteria of genus fusobacterium and use of same

Abstract

The present invention addresses the problem of controlling bacteria of the genus Fusobacterium by a method which is considered to have less impact on beneficial bacteria. The present invention provides a composition for controlling bacteria belonging to the genus Fusobacterium in a digestive tract, particularly in the oral cavity or intestinal flora, the composition containing any one substance selected from the group consisting of a flavonoid that releases a compound represented by formula I in the digestive tract, a transglycosylation product thereof, and a compound represented by formula I.

Description

Composition for controlling Fusobacterium genus bacteria and its use

[0001] The present invention relates to a composition for controlling bacteria of the genus Fusobacterium and uses thereof.

[0002] Fusobacterium is a bacterium present in the human oral cavity and intestines and is known to be a causative agent of periodontal disease. Fusobacterium nucleatum is also known to cause opportunistic infections and has recently been implicated in colon cancer (e.g., Non-Patent Document 1). Furthermore, F. nucleatum has been isolated from gastrointestinal infections such as appendicitis (Non-Patent Document 2), and its association with inflammatory bowel disease has recently been suggested (Non-Patent Documents 3, 4, 5). Oral Fusobacterium nucleatum is known to cause periodontal disease, halitosis, oral cancer, esophageal cancer, aspiration pneumonia, and Alzheimer's disease (Non-Patent Documents 6, 7, 8, 9, 10).

[0003] On the other hand, hesperidin, which is classified as a flavanone, a type of flavonoid, is known to have several effects. For example, Patent Document 1 describes a cancer inhibitor containing hesperidin glycoside. Patent Document 2 describes a functional sweetener composition containing at least one anti-inflammatory agent, at least one high-intensity sweetener, and at least one sweetness-improving composition, and hesperidin is listed as an example of the anti-inflammatory agent. Furthermore, Patent Document 3 describes a food composition for enhancing at least one effect selected from the group consisting of anti-cancer effect, anti-muscle atrophy effect, anti-obesity effect, cholesterol-lowering effect, anti-thrombosis effect, and immune-enhancing effect of a green tea extract or catechin (first component), containing a citrus extract or a flavanone or its glycoside (second component), and the second component is at least one selected from the group consisting of eriodictyol, naringenin, hesperetin, and their glycosides. Non-Patent Document 11 describes the effects of hesperidin, hesperetin, and narigenin on intestinal barrier function and inflammation.

[0004] Furthermore, Non-Patent Document 12 presents the measured minimum inhibitory concentrations and minimum bactericidal concentrations of hesperidin, hesperetin, and naringenin against F. nucleatum, a causative bacterium of periodontitis. Patent Document 4 describes a composition for suppressing bad breath containing (A) at least one selected from the group consisting of citric acid and its salts, and (B) at least one selected from the group consisting of hesperidin, nomilin, and limonin, and presents experimental results showing that the addition of lemon juice containing citric acid, hesperidin, and other ingredients reduced the number of F. nucleatum bacteria in saliva compared to the addition of water. Patent Document 5 describes an agent for improving intestinal flora containing polyphenols and polyphenol oxidase as active ingredients, and presents experimental results showing that the occupancy rate of Fusobacterium bacteria in feces was reduced when sample A containing coffee extract (a plant extract containing polyphenols) and burdock freeze-dried powder (a plant freeze-dried product containing polyphenol oxidase) was ingested.

[0005] Recently, research has been conducted to identify the target of hesperidin in cancer (Non-Patent Document 13), and the effects of hesperidin on cardiovascular disease risk factors and the role of the intestinal flora in hesperidin bioavailability have been reported (Non-Patent Document 14).

[0006] JP 2003-40781 A WO2007 / 061912 (JP 2009-517044) WO2015 / 199169 (Patent No. 6912763) JP 2020-200245 A WO2022-138698 A

[0007] C. A. Brennan et ai. Fusobacterium nucleatum - symbiont, opportunist and oncobacterium. Nat Rev Microbiol. 2019;17(3):156-166. doi: 10.1038 / s41579-018-0129-6.Metzger Z et al. Synergistic pathogenicity of Porphyromonas gingivalis and Fusobacterium nucleatum in the mouse subcutaneous chamber model. J. Endod 35, 86-94 (2009). [PubMed: 19084132]Strauss J et al. Invasive potential of gut mucosa-derived Fusobacterium nucleatum positively correlates with IBD status of the host. Inflamm. Bowel Dis 17, 1971-1978 (2011). [PubMed: 21830275]Gevers D et al. The treatment-naive microbiome in new-onset Crohn’s disease. Cell Host Microbe 15, 382-392 (2014). [PubMed: 24629344]Pascal V et al. A microbial signature for Crohn’s disease. Gut 66, 813-822 (2017). [PubMed: 28179361]Sakanaka A et al. Fusobacterium nucleatum Metabolically Integrates Commensals and Pathogens in Oral Biofilms. mSystems 2022 Aug 30;7(4):e0017022.Wenhua Shao et al.Conversion from epithelial to partial-EMT phenotype by Fusobacterium nucleatum infection promotes invasion of oral cancer cells. Sci Rep. 2021 Jul 22;11(1):14943.Yamamura K et al., Human Microbiome Fusobacterium Nucleatum in Esophageal Cancer Tissue Is Associated with Prognosis. Clin Cancer Res. 2016 Nov 15;22(22):5574-5581.SUZUKI R et al. Periodontopathic Bacterium Fusobacterium nucleatum Affects Matrix Metalloproteinase-9 Expression in Human Alveolar Epithelial Cells and Mouse Lung. In Vivo March 2022, 36 (2) 649-656.Hongle Wu et al. The Periodontal Pathogen Fusobacterium nucleatum Exacerbates Alzheimer’s Pathogenesis via Specific Pathways. Front Aging Neurosci. 2022; 14: 912709.Nutrients 2019, 11(7), 1464Food & Function (2015), 6(3), 719-729Vaishali Aggarwal et al. Molecular mechanisms of action of hesperidin in cancer: Recent trends and advancements. Exp Biol Med (Maywood). 2020 Mar; 245(5): 486-497.Anna Mas-Capdevila et al.Effect of Hesperidin on Cardiovascular Disease Risk Factors: The Role of Intestinal Microbiota on Hesperidin Bioavailability. Nutrients. 2020 May 20;12(5):1488. doi: 10.3390 / nu12051488.

[0008] In the digestive tract, especially in the oral cavity and intestines, Fusobacterium bacteria exist in a complex symbiotic system with other bacteria, making it difficult to control. For example, the administration of disinfectants and antibiotics is thought to reduce not only Fusobacterium bacteria but also beneficial bacteria. Therefore, control using probiotics and prebiotics, which do not affect beneficial bacteria, is desired.

[0009] No effective material has been found to control Fusobacterium.

[0010] The present invention provides the following: [1] A composition for controlling bacteria belonging to the genus Fusobacterium in the gastrointestinal tract, comprising any one selected from the group consisting of a flavonoid that liberates a compound represented by formula I in the gastrointestinal tract, a glycosyltransfer thereof, and the compound represented by formula I. (wherein R1 to R4 are the following combinations): [2] A composition for controlling bacteria belonging to the genus Fusobacterium, comprising any one selected from the group consisting of a flavonoid and its glycosyltransferase that releases a compound represented by formula I upon hydrolysis with glucosidase, and the compound represented by formula I. (wherein R1 to R4 are the following combinations): [3] The composition according to 1 or 2, wherein R1 to R4 are the following combinations: [4] The composition according to any one of items 1 to 3, wherein the compound represented by formula I is hesperetin, the flavonoid that liberates hesperetin is hesperidin, and the composition comprises any one selected from the group consisting of hesperidin, a glycosyltransfer of hesperidin, and hesperetin. [5] The composition according to any one of items 1 to 4, wherein the glycosyltransfer of hesperidin is glucosylhesperidin. [6] A composition for controlling bacteria belonging to the genus Fusobacterium, comprising hesperetin. [7] The composition according to any one of items 1 to 6, wherein the control of bacteria belonging to the genus Fusobacterium is a reduction in the number of bacteria belonging to the genus Fusobacterium. [8] The composition according to any one of items 1 to 7, for use as any one selected from the group consisting of prebiotics and synbiotics. [9] The composition according to any one of 1 to 8 for treating any one selected from inflammatory bowel disease, colon cancer, periodontal disease, halitosis, oral cancer, esophageal cancer, aspiration pneumonia, and Alzheimer's dementia through control of bacteria belonging to the genus Fusobacterium, or the composition according to any one of claims 1 to 8 for treating any one selected from inflammatory bowel disease, colon cancer, periodontal disease, halitosis, oral cancer, esophageal cancer, aspiration pneumonia, and Alzheimer's dementia in a subject in whom Fusobacterium bacteria have been detected in at least one of the intestinal tract and feces.

[10] The composition according to any one of 1 to 8 for treating any one selected from inflammatory bowel disease, colon cancer, periodontal disease, halitosis, oral cancer, esophageal cancer, aspiration pneumonia, and Alzheimer's dementia.

[11] The composition according to any one of items 1 to 9, wherein the digestive tract is the oral cavity, or the composition according to item 1, wherein the digestive tract is the oral cavity, contains as an active ingredient any one selected from the group consisting of hesperetin and transglycosylation products of hesperidin.

[12] The composition according to any one of items 1 to 9, wherein the digestive tract is the intestine.

[13] A food composition, which is any one selected from the group consisting of a flavonoid that liberates a compound represented by formula I in the digestive tract and a transglycosylation product thereof, as defined in item 1, and a compound represented by formula I (excluding transglycosylation hesperidin), and is selected from the group consisting of a food with functional claims, a food for specified health use, a fermented milk, a milk beverage, and a powdered food.

[14] A pharmaceutical composition comprising any one selected from the group consisting of a flavonoid and a transglycosylation product thereof that liberates a compound represented by formula I in the gastrointestinal tract, and a compound represented by formula I, as defined in 1, and a pharmaceutically acceptable excipient.

[15] An enteric-coated formulation comprising any one selected from the group consisting of a flavonoid and a transglycosylation product thereof that liberates a compound represented by formula I in the gastrointestinal tract, as defined in 1, and a compound represented by formula I.

[16] The composition according to 13 or 14, or the formulation according to 15, containing 0.01 g or more and 7 g or less of any one selected from the flavonoid and a transglycosylation product thereof, and the compound.

[0011]

[21] A method for controlling bacteria belonging to the genus Fusobacterium in the intestine of a subject, or a method for reducing the risk of at least one of inflammatory bowel disease and colorectal cancer in a subject, comprising the following steps: having the subject ingest a composition or a food material comprising any one selected from the group consisting of a flavonoid and a transglycosylation product thereof that liberates a compound represented by formula I defined above (preferably hesperetin) in the intestine, and the compound represented by formula I defined above (preferably hesperetin), based on the presence or amount of bacteria belonging to the genus Fusobacterium in the intestine of the subject.

[22] A method for suggesting to a subject the intake of a composition, a method for supporting a subject's diet, or a method for supporting a subject's health, comprising the following steps: suggesting to a subject the intake of a composition or a food material comprising any one selected from the group consisting of a flavonoid and a transglycosylation product thereof that liberates a compound represented by formula I defined above (preferably hesperetin) in the intestine, and the compound represented by formula I defined above (preferably hesperetin), based on the presence or amount of bacteria belonging to the genus Fusobacterium in the intestine of the subject.

[23] A method for controlling bacteria belonging to the genus Fusobacterium in the oral cavity of a subject, or a method for reducing the risk of any one selected from periodontal disease, halitosis, oral cancer, esophageal cancer, aspiration pneumonia, and Alzheimer's disease in a subject, comprising the following steps: based on the presence or amount of bacteria belonging to the genus Fusobacterium in the oral cavity of the subject, having the subject ingest or gargle with a composition or food material containing any one selected from the group consisting of flavonoids and transglycosylation products thereof that liberate a compound represented by formula I defined above (preferably hesperetin) in the oral cavity, and the compound represented by formula I defined above (preferably hesperetin).

[24] A method for suggesting to a subject that they take a composition or gargle, a method for supporting the subject's dietary habits, or a method for supporting the health of a subject, the method comprising the following steps: a step of suggesting to a subject that they take a composition or food material containing any one selected from the group consisting of flavonoids and transglycosylation products thereof that liberate a compound represented by formula I defined above (preferably hesperetin) in the oral cavity, and the compound represented by formula I defined above (preferably hesperetin), based on the presence or amount of bacteria belonging to the genus Fusobacterium in the oral cavity of the subject.

[0012]

[31] A composition for use in a method for controlling bacteria belonging to the genus Fusobacterium in the gastrointestinal tract, comprising any one selected from the group consisting of a flavonoid (preferably hesperetin) that liberates the compound represented by formula I defined above in the gastrointestinal tract and a transglycosylation product thereof, and the compound represented by formula I defined above (preferably hesperetin). Use of any one selected from the group consisting of a flavonoid (preferably hesperetin) that liberates the compound represented by formula I defined above in the gastrointestinal tract and a transglycosylation product thereof, and the compound represented by formula I defined above (preferably hesperetin), in the manufacture of a composition for controlling bacteria belonging to the genus Fusobacterium in the gastrointestinal tract. A method or non-therapeutic method for controlling bacteria belonging to the genus Fusobacterium in the gastrointestinal tract, comprising the step of administering to a subject a composition comprising any one selected from the group consisting of a flavonoid (preferably hesperetin) that liberates the compound represented by formula I defined above in the gastrointestinal tract and a transglycosylation product thereof, and the compound represented by formula I defined above (preferably hesperetin). Use or non-therapeutic use of a composition comprising any one selected from the group consisting of a flavonoid (preferably hesperetin) and a transglycosylation product thereof that liberates the compound represented by formula I defined above in the gastrointestinal tract, and the compound represented by formula I defined above (preferably hesperetin), for controlling bacteria belonging to the genus Fusobacterium in the gastrointestinal tract.

[32] A composition comprising any one selected from the group consisting of a flavonoid (preferably hesperetin) and a transglycosylation product thereof that liberates the compound represented by formula I defined above upon hydrolysis with a glucosidase, and the compound represented by formula I defined above (preferably hesperetin), for use in a method for controlling bacteria belonging to the genus Fusobacterium. Use of any one selected from the group consisting of a flavonoid (preferably hesperetin) and a transglycosylation product thereof that liberates the compound represented by formula I defined above upon hydrolysis with a glucosidase, and the compound represented by formula I defined above (preferably hesperetin), in the manufacture of a composition for controlling bacteria belonging to the genus Fusobacterium.A method or non-therapeutic method for controlling bacteria belonging to the genus Fusobacterium, comprising the step of administering to a subject a composition comprising any one selected from the group consisting of a flavonoid (preferably hesperetin) and a trans-glycosylation product thereof that liberates the compound represented by formula I defined above upon hydrolysis with a glucosidase, and a compound represented by formula I defined above (preferably hesperetin). Use or non-therapeutic use of a composition comprising any one selected from the group consisting of a flavonoid (preferably hesperetin) and a trans-glycosylation product thereof that liberates the compound represented by formula I defined above upon hydrolysis with a glucosidase, and a compound represented by formula I defined above (preferably hesperetin), for controlling bacteria belonging to the genus Fusobacterium.

[33] The composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to 31 or 32, wherein R1 to R4 are any of the following combinations:

[34] The composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to any one of items 31 to 33, wherein the compound represented by formula I is hesperetin and the flavonoid that liberates hesperetin is hesperidin, and the composition comprises any one selected from the group consisting of hesperidin, a transglycosyl hesperidin, and hesperetin.

[35] The composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to any one of items 31 to 34, wherein the transglycosyl hesperidin is glucosylhesperidin.

[36] A composition comprising hesperetin for use in a method for controlling bacteria belonging to the genus Fusobacterium. Use of hesperetin in the production of a composition for controlling bacteria belonging to the genus Fusobacterium. A method or non-therapeutic method for controlling bacteria belonging to the genus Fusobacterium, comprising the step of administering a composition comprising hesperetin to a subject.

[37] The composition according to any one of items 31 to 35, wherein the control of bacteria belonging to the genus Fusobacterium is a reduction in the number of bacteria belonging to the genus Fusobacterium.

[38] The composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to any one of items 31 to 36, wherein the composition is for use as any selected from the group consisting of prebiotics and synbiotics.

[39] The composition, use in manufacturing, method or non-therapeutic method, or use or non-therapeutic use according to any one of items 31 to 38, wherein the composition is for treating any one selected from inflammatory bowel disease, colon cancer, periodontal disease, halitosis, oral cancer, esophageal cancer, aspiration pneumonia, and Alzheimer's dementia via control of bacteria belonging to the genus Fusobacterium; or the composition, use in manufacturing, method or non-therapeutic method, or use or non-therapeutic use according to any one of items 31 to 38, wherein the composition is for treating any one selected from inflammatory bowel disease, colon cancer, periodontal disease, halitosis, oral cancer, esophageal cancer, aspiration pneumonia, and Alzheimer's dementia in a subject in whom Fusobacterium bacteria have been detected in at least one of the intestinal tract and stool.

[40] The composition, use in manufacturing, method or non-therapeutic method, or use or non-therapeutic use according to any one of items 31 to 38, which is for the treatment of any one selected from inflammatory bowel disease, colon cancer, periodontal disease, halitosis, oral cancer, esophageal cancer, aspiration pneumonia, and Alzheimer's dementia.

[41] The composition, use in manufacturing, method or non-therapeutic method, or use or non-therapeutic use according to any one of items 31 to 40, wherein the digestive tract is the oral cavity, or the composition, use in manufacturing, method or non-therapeutic method, or use or non-therapeutic use according to any one of items 31 to 40, wherein the composition comprises as an active ingredient any one selected from the group consisting of hesperetin and transglycosylation products of hesperidin, and wherein the digestive tract is the oral cavity.

[42] The composition, use in manufacturing, method or non-therapeutic method, or use or non-therapeutic use according to any one of items 31 to 40, wherein the digestive tract is the intestine.

[0013] Using any one selected from the group consisting of flavonoids and their glycosyltransferases that release a compound represented by formula I in the gastrointestinal tract, and the compound represented by formula I, it is possible to provide a means for controlling bacteria belonging to the genus Fusobacterium in the gastrointestinal flora. Controlling bacteria belonging to the genus Fusobacterium in the intestinal flora is expected to lead to the treatment of colon cancer and inflammatory bowel disease. Controlling bacteria belonging to the genus Fusobacterium in the oral flora is expected to lead to the treatment of periodontal disease, halitosis, esophageal cancer, aspiration pneumonia, and Alzheimer's disease.

[0014] Number of Fusobacterium bacteria after cultivation *P <0.05 versus Control, Wilcoxon rank sum test Number of Fusobacterium colonies after cultivation

[0015] [Active Ingredient] The present invention relates to a composition for controlling bacteria of the genus Fusobacterium, comprising as an active ingredient any one selected from the group consisting of flavonoids and their transglycosylation products that release a compound represented by formula I in the digestive tract, preferably the oral cavity or intestine, and the compound represented by formula I. The present invention also relates to a composition for controlling bacteria belonging to the genus Fusobacterium, more particularly, for controlling bacteria belonging to the genus Fusobacterium in the presence of glucosidase, comprising any one selected from the group consisting of flavonoids and their transglycosylation products that release a compound represented by formula I upon hydrolysis with glucosidase, and the compound represented by formula I. "Any one" refers to any number and type. The composition may contain only one active ingredient or multiple active ingredients. "Containing as an active ingredient" refers to the use in the composition of an effective amount to exert the intended function, or the use of an ingredient that contributes to the intended purpose and is identified in the label. In functional food products, active ingredients are sometimes referred to as functional ingredients (ingredients that contribute to specific health purposes (excluding those related to reducing disease risk)).

[0016] (Compound represented by formula I and flavonoid liberating the same) In one embodiment, the composition contains, as an active ingredient, any one selected from the group consisting of a flavonoid that liberates a compound represented by formula I in the digestive tract, preferably in the oral cavity or intestine, and the compound represented by formula I. Flavonoid is a general term for polyphenolic compounds synthesized by plants. Flavonoids may be glycosides. The compound represented by formula I may be an aglycone.

[0017] In the formula, R1 to R4 are the following combinations:

[0018]

[0019] The release of the compound represented by formula I in the digestive tract refers to the production of the compound represented by formula I by the action of an enzyme produced by bacteria in the digestive tract or an enzyme secreted into the digestive tract. The enzyme may be an oxidoreductase, transferase, hydrolase, lyase, isomerase, ligase, or ABC transporter.

[0020] In one embodiment, R1 to R4 are the following combinations: That is, the compound represented by formula I can be hesperetin, naringenin, or eriodictyol.

[0021]

[0022] In a particularly preferred embodiment, the compound of formula I is hesperetin and the flavonoid that liberates hesperetin is hesperidin.

[0023] Hesperidin (molecular formula: C 28 H 34 O 15 Hesperetin (molecular formula: C) is a polyphenol known as vitamin P. 16 H 14 Hesperidin is a glycoside of β-rutinose (6-O-α-L-rhamnosyl-D-β-glucose). It has very low solubility in water (approximately 0.002 g / 100 mL).

[0024] In the following, the present invention and its embodiments will be explained using hesperetin and hesperidin as examples, but those skilled in the art will be able to understand the explanation by applying it to the compound represented by formula I and the flavonoids that liberate it.

[0025] In the intestine, hesperidin is acted upon by β-glucosidase to liberate hesperetin (Non-Patent Document 14). Because not only humans but many intestinal bacteria possess β-glucosidase, it is thought that most humans possess bacteria with β-glucosidase in their intestines (Marta Dabek et al., Deglycosylation of flavonoid and isoflavonoid glycosides by human small intestine and liver β-glucosidase activity. FEMS Microbiol Ecol. 2008 Dec;66(3):487-95. doi: 10.1111 / j.1574-6941.2008.00520.x.).

[0026] In the oral cavity, hesperidin is released into hesperetin when β-glucosidase produced by oral bacteria acts on it. Because many oral bacteria possess β-glucosidase, it is thought that most humans possess bacteria that possess β-glucosidase in their oral cavity. β-glucosidase enzyme activity has also been reported in human saliva (Lucimari Teixeira Essenfelder et al. Salivary β-glucosidase as a direct factor influencing the occurrence of halitosis. Biochem Biophys Rep. 2021 Mar 6;26:100965.).

[0027] Flavonoids that release various compounds represented by formula I and the compounds represented by formula I can be obtained by extraction from plants. Preferred examples of plants from which hesperidin and hesperetin can be obtained are citrus fruits, more preferred examples being mandarin oranges, unshu mandarin oranges, yuzu citrus, oranges, tankan oranges, ponkan oranges, sudachi, shikwasa, and lemons. They are preferably obtained from the fruits, peels, and unripe fruits of these fruits. The composition may contain the flavonoids that release the compounds represented by formula I and the compounds represented by formula I as plant extracts.

[0028] (Glycosyltransferate, Monoglucosyl Hesperidin) In one embodiment, the composition comprises a flavonoid transglycosyltransferate that releases the compound represented by formula I in the digestive tract, preferably in the oral cavity or intestine. A flavonoid transglycosyltransferate refers to a flavonoid to which 1 to 20 monosaccharides have been added. A flavonoid transglycosyltransferate is usually more water-soluble than the parent flavonoid itself.

[0029] In one embodiment, the composition contains a transglycosylation product of hesperidin (also referred to as transglycosyl hesperidin) as a transglycosylation product of a flavonoid that liberates hesperetin in the digestive tract, preferably in the oral cavity or intestines. Examples of transglycosylation hesperidins include α-glucosyl hesperidin (also referred to as monoglucosyl hesperidin), α-diglucosyl hesperidin, α-triglucosyl hesperidin, α-tetraglucosyl hesperidin, and α-pentaglucosyl hesperidin.

[0030] In a preferred embodiment, the composition contains monoglucosyl hesperidin as a glycosyltransferase of a flavonoid that liberates hesperetin in the digestive tract, preferably in the oral cavity or intestine.

[0031] Monoglucosyl hesperidin (molecular weight: 772.70) has a higher solubility in water than hesperidin (approximately 197 g / 100 mL of water). Monoglucosyl hesperidin is broken down by α-glucosidase and β-glucosidase to produce hesperidin. α-glucosidase is contained in saliva and is also secreted by intestinal bacteria. β-glucosidases have been identified in humans, intestinal bacteria, and oral bacteria (AJ Day at al. Deglycosylation of flavonoid and isoflavonoid glycosides by human small intestine and liver β-glucosidase activity. FEBS Lett. 25;436(1):71-5, 2019, Matsumoto S. et al. Glucosyl hesperidin: safety studies. Fundam Toxicol Sci 8:299-317, 2019).

[0032]

[0033] Flavonoid transglycosylation products are usually produced by enzymatically transferring sugars from a glycosyl donor to a flavonoid. Japanese Patent Application Laid-Open No. 7-107972 can be referenced for transglycosylation products of hesperidin, eriocitrin, naringin, neohesperidin, diosmin, linarin, poncirin, prunin, or hesperetin-7-glucoside and their production methods. Japanese Patent Application Laid-Open No. 3-7593 can be referenced for transglycosylation products of hesperidin and their production methods.

[0034] [Uses] (Function / Action) The composition of the present invention can be used to control bacteria belonging to the genus Fusobacterium (sometimes simply referred to as Fusobacterium bacteria). In a preferred embodiment, the composition can be used to control Fusobacterium bacteria in the digestive tract, preferably in the oral or intestinal flora. Controlling Fusobacterium bacteria in the oral or intestinal flora refers to controlling bacteria belonging to the genus Fusobacterium in a state where various bacteria living in the oral cavity or intestine are present. Controlling bacteria refers to controlling the number of bacteria, regulating the intestinal flora including Fusobacterium bacteria, or assisting in regulating the intestinal flora including Fusobacterium bacteria. In a preferred embodiment, the composition is used to reduce the number of Fusobacterium bacteria or to assist in reducing the number of Fusobacterium bacteria.

[0035] The oral cavity includes the teeth, periodontal tissue, tongue, and salivary glands. The intestine refers to the digestive tract in humans and animals where bacteria reside and where ingested food is digested and absorbed. The intestine (sometimes called the intestinal tract) includes the small intestine and large intestine, preferably the large intestine.

[0036] Examples of Fusobacterium genus bacteria include: Fusobacterium nucleatum subsp. nucleated, Fusobacterium nucleatum subsp. animal, Fusobacterium nucleatum subsp. vincentius (fusiform), Fusobacterium nucleatum subsp. polymorphic, Fusobacterium periodonticum, Fusobacterium russii, Fusobacterium canifelinum, Fusobacterium simiae, Fusobacterium necrophorum, Fusobacterium equinus, Fusobacterium gonidiaformans, Fusobacterium deadly, Fusobacterium necrogenes, Fusobacterium ulcerans, Fusobacterium varium, Fusobacterium perfoetens

[0037] In the present invention, the term "Fusobacterium" refers to bacteria identified as belonging to the genus Fusobacterium by molecular phylogenetic analysis based on the 16S rRNA gene. Criteria for determining genera based on molecular phylogenetic analysis based on the 16S rRNA gene are well known to those skilled in the art (Stackebrandt E, Ebers J. Taxonomic parameters revisited: tarnished gold standards. Microbiol Today 2006;33:152-155).

[0038] Whether a certain component controls Fusobacterium can be evaluated as follows. Specifically, an appropriate Fusobacterium (e.g., Fusobacterium nucleatum JCM JCM8532T) is inoculated onto GAM agar media containing and not containing the component of interest, and the media is cultured under appropriate conditions (e.g., 37°C, anaerobic conditions, 48 ​​hours), and the colony counts are counted and compared. The results are then compared with those for agar media cultured under identical conditions except for the addition of a control (e.g., sterilized water) instead of the component of interest.

[0039] Whether a certain component controls Fusobacterium bacteria in the intestinal flora can be evaluated as follows: Feces provided by a healthy individual is added to an appropriate medium and, if necessary, cultured for a certain period of time. After that, the component to be evaluated is added and cultured under appropriate conditions (e.g., 16 hours of culture at 37°C under anaerobic conditions, similar to the conditions in the intestinal tract). If the feces does not contain Fusobacterium bacteria, any available Fusobacterium bacteria is added at the start of culture and culture is continued. An example of the Fusobacterium bacteria to be added is F. nucleatum JCM8532T. The amount added can be adjusted appropriately depending on the experimental conditions, but can be, for example, 0.1 to 10 × 10 4 The number of Fusobacterium bacteria in the bacterial flora of the culture is then measured. The results can then be compared with those of a culture grown under identical conditions except that a control (e.g., sterile water) was added instead of the component to be evaluated, and a judgment can be made.

[0040] The number of Fusobacterium bacteria contained in the bacterial flora can be determined by known methods. One preferred method is to perform 16S metagenomic analysis (sequence analysis of 16S rRNA gene amplicons) on DNA extracted from the culture. When performing 16S metagenomic analysis, DNA extraction can be performed using a commercially available kit. The genomic region to be analyzed is not particularly limited as long as it allows identification of the bacteria, but the V3-V4 region of the 16S rRNA gene can be used. Primers, amplification conditions, amplicon purification, and other methods for bacterial analysis are also well known to those skilled in the art. Sequence analysis is preferably performed using a next-generation sequencer with higher performance. QIIME 2 is used to analyze the obtained data. TM For 16S metagenomic analysis, those skilled in the art can refer to information such as Sanschagrin S, Yergeau E. Next-generation sequencing of 16S ribosomal RNA gene amplicons. J Vis Exp. 2014;(90):51709. Published 2014 Aug 29. doi:10.3791 / 51709.

[0041] The Fusobacterium bacteria of the present invention are bacteria present in the human oral cavity and intestinal tract and are known to be the causative bacteria of periodontal disease. Fusobacterium nucleatum is also known to cause opportunistic infections and has recently been implicated in colon cancer (e.g., Non-Patent Document 1). Furthermore, F. nucleatum has been isolated from gastrointestinal infections such as appendicitis (Non-Patent Document 2), and its association with inflammatory bowel disease has recently been suggested (Non-Patent Documents 3, 4, and 5). Therefore, in one embodiment, the composition can be used for the treatment of any of inflammatory bowel disease and colon cancer via the control of Fusobacterium bacteria.

[0042] Inflammatory bowel diseases include ulcerative colitis and Crohn's disease.

[0043] Fusobacterium nucleatum in the oral cavity is known to cause periodontal disease, halitosis, oral cancer, esophageal cancer, aspiration pneumonia, and Alzheimer's dementia (Non-Patent Documents 6, 7, 8, 9, and 10). Therefore, in one embodiment, the composition can be used for periodontal disease, halitosis, oral cancer, esophageal cancer, aspiration pneumonia, and Alzheimer's dementia. More specifically, the composition can be used for periodontal disease, halitosis, oral cancer, esophageal cancer, aspiration pneumonia, and Alzheimer's dementia through control of Fusobacterium bacteria.

[0044] Furthermore, the composition can be used for treating a variety of diseases or conditions that can be improved by controlling the proliferation of Fusobacterium bacteria in the intestinal tract.

[0045] In the present invention, the term "treatment" for a disease or condition includes reducing the risk of onset, delaying onset, prevention, cure, and halting or delaying progression. It also includes helping to improve from a disease or condition, alleviating a disease or condition, and reducing the risk of developing a disease or condition. Treatment includes radical treatment (treatment that eliminates the cause) and symptomatic treatment (treatment that improves symptoms). Treatment for improvement or treatment includes medical procedures performed by physicians and nurses and midwives under the direction of physicians, as well as non-therapeutic procedures performed by persons other than physicians, such as pharmacists, nutritionists (including registered dietitians and sports nutritionists), public health nurses, midwives, nurses, clinical laboratory technicians, sports instructors, pharmaceutical manufacturers, pharmaceutical distributors, food manufacturers, and food distributors. Prevention or reduction of the risk of onset also includes recommending the intake of specific foods and providing nutritional guidance (including nutritional guidance necessary for the recovery of injured or sick individuals, and nutritional guidance for maintaining and promoting health).

[0046] (Subject) The composition of the present invention is suitable for administration to subjects for whom it is desirable or necessary to control Fusobacterium bacteria in the digestive tract, preferably in the oral cavity or intestinal flora, particularly to reduce the number of bacteria; subjects selected from the group consisting of inflammatory bowel disease or colon cancer; subjects with a disease or condition that can be improved by controlling the proliferation of Fusobacterium bacteria in the intestinal tract; subjects with a disease or condition that can be improved by controlling the proliferation of Fusobacterium bacteria in the oral cavity; subjects with a disease or condition that can be improved by controlling the proliferation of Fusobacterium bacteria in the oral cavity. In the present invention, the term "administer" is used not only to refer to administering a pharmaceutical product to a subject, but also to refer to having a subject ingest a non-pharmaceutical product such as a food product. "Administer" can be interpreted as "intake" (or "to make someone take"), and "administer" can be interpreted as "administered".

[0047] Subjects for whom it is desirable or necessary to control Fusobacterium bacteria in the intestinal tract include subjects in whom Fusobacterium bacteria have been detected in at least one of their intestinal tract and feces, and subjects in whom relatively high numbers of Fusobacterium bacteria have been detected in at least one of their intestinal tract and feces. Subjects for whom it is desirable or necessary to control Fusobacterium bacteria in the intestinal tract also include subjects who have been found to have relatively high numbers of Fusobacterium bacteria by any test or analysis, such as an intestinal flora test, a test for substances in feces, or fecal metabolome analysis, subjects in whom Fusobacterium bacteria have been detected, subjects who have been found to be at high risk of inflammatory bowel disease, subjects who have been found to be at high risk of colorectal cancer, and subjects who have been recommended, based on the test or analysis results, by a doctor, nurse, pharmacist, nutritionist, etc., or by a computer program, to consume foods, beverages, or medicines that can control Fusobacterium bacteria.

[0048] Subjects for whom it is desirable or necessary to control Fusobacterium bacteria in the oral cavity include those in whom Fusobacterium bacteria have been detected in the oral cavity (e.g., any one selected from saliva, tongue coating, and dental plaque), and those in whom relatively high numbers of Fusobacterium bacteria have been detected in any one selected from saliva, tongue coating, and dental plaque. Furthermore, subjects for whom it is desirable or necessary to control Fusobacterium in the oral cavity include subjects who have been found to have a relatively high number of Fusobacterium bacteria through any test or analysis, such as an oral flora test, a test for substances contained in a sample selected from saliva, tongue coating, and dental plaque, or metabolomic analysis of a sample selected from saliva, tongue coating, and dental plaque, subjects in which Fusobacterium bacteria have been detected, subjects who have been found to be at high risk of periodontal disease, subjects who have been found to be at high risk of developing halitosis, subjects who suffer from halitosis, and subjects who have been recommended, based on the test or analysis results, by a doctor, nurse, pharmacist, nutritionist, etc., or by a computer program, to consume foods, beverages, or medicines that can control Fusobacterium bacteria. Also included are subjects who wish to reduce the number of Fusobacterium bacteria in at least one of their intestines and feces, subjects who wish to reduce their risk of colorectal cancer, and subjects who wish to reduce their risk of inflammatory bowel disease. In one embodiment, the subject is a subject in which Fusobacterium bacteria have been detected in at least one of the intestines and feces, or a subject in which the number of Fusobacterium bacteria has been found to be relatively high. The term "a subject in which the number of Fusobacterium bacteria has been detected in at least one of the intestines and feces" means, for example, that when feces collected from a subject is cultured at a concentration of 0.1% (w / v) under anaerobic conditions at 37°C for 16 hours, the number of Fusobacterium bacteria in the culture medium is 1 x 10 3 cells / mL or more, preferably 1 x 10 4 cells / mL or more, preferably 1×10 4 cells / mL or more, more preferably 5 x 10 5 cells / mL or more, or the number of Fusobacterium bacteria in the feces collected from the subject is 1 x 10 4cells / g or more, preferably 5×10 4 cells / g or more, preferably 1×10 5 cells / g or more, more preferably 5×10 5 This refers to the case where the number of bacteria is more than 10 ...

[0049] In one embodiment, the subject is healthy (not having a medically diagnosed illness). The subject may be a human or a non-human animal.

[0050] The subject's age is not particularly limited, and the subject may be, for example, a newborn (within 28 days of birth); an infant (less than 1 year of age); a toddler (1-6 years of age); a child (7 years or older, but younger than 15 years of age); an adult (15 years or older); or a person 65 years or older. In a preferred embodiment, the subject is a relatively elderly person, such as a person 60 years or older, 65 years or older, 70 years or older, or 75 years or older. It has been reported that the number of Fusobacterium bacteria in the intestinal tract increases with age (e.g., Odamaki et al. BMC Microbiology (2016) 16:90 DOI 10.1186 / s12866-016-0708-5). It has also been shown that saliva containing Fusobacterium bacteria in the oral cavity can reach the lungs via the trachea, causing lung infection and aspiration pneumonia.

[0051] [Composition] (Food composition, etc.) The composition of the present invention can be a food composition or a pharmaceutical composition. Foods and pharmaceuticals include not only those for humans but also those for non-human animals, unless otherwise specified. Foods include general foods, functional foods, nutritional compositions, and also therapeutic foods (those that serve the purpose of treatment; prepared based on a menu prepared by a nutritionist or the like in accordance with a doctor's dietary prescription), dietary therapy foods, ingredient-modified foods, nursing care foods, and foods for medical support, unless otherwise specified. Foods include not only solid foods but also liquid foods, such as beverages, energy drinks, liquid foods, and soups, unless otherwise specified. Functional foods refer to foods that can impart a specific functionality to the body, and include, for example, foods for specified health uses (including conditional FOSHUs [foods for specified health uses]), foods with functional claims, health functional foods including foods with nutrient function claims, foods for special dietary uses, dietary supplements, health supplements, supplements (e.g., tablets, coated tablets, sugar-coated tablets, enteric-coated preparations, enteric-coated preparation capsules (e.g., enteric-coated soft capsules, enteric-coated hard capsules, colon delivery capsules), liquid preparations, and various other dosage forms), dietary supplements, food supplements, medical foods (as defined by the U.S. Food and Drug Administration (FDA)), beauty foods (e.g., diet foods), and other health foods in general. Furthermore, in the present invention, "functional foods" also includes health foods to which a health claim based on the Codex Alimentarius (Joint FAO / WHO Food Standards Commission) is applied. A food supplement is a concentrated form of nutrients or other substances, either alone or in combination, that supplements the normal diet and has a nutritional or physiological effect, and is labeled as a food supplement. A dietary supplement is a product (other than tobacco) intended to supplement the diet, contains one or more of the ingredients in question, and is labeled as a dietary supplement.

[0052] The composition may be for dental or oral use. The composition for dental or oral use may be a pharmaceutical composition or a non-pharmaceutical composition such as a quasi-drug.

[0053] In one embodiment, a food material can be used instead of the food composition. In the present invention, the food material refers to all or part of an unprocessed plant. Preferred examples of the food material are citrus fruits (Rutaceae, Rutaceae, Citrus family), more preferably fruits, peels, and unripe fruits such as mandarin oranges, unshiu mandarins, oranges, tankan oranges, ponkan oranges, sudachi citrus, shiikuwasa lemons, lemons, kabosu citrus, and kumquats.

[0054] (Route of Administration, etc.) The composition of the present invention may be administered orally, parenterally, for example, via a tube (gastrostomy, enterostomy), or nasally, but is preferably administered orally.

[0055] The composition can be administered to a subject repeatedly or continuously for a long period of time. The period is not particularly limited, but to ensure sufficient efficacy, it is preferable to administer the composition continuously for a relatively long period of time, for example, 3 days or more, 1 week or more, 2 weeks or more, 1 month or more, 3 months or more, 6 months or more, or 1 year or more.

[0056] The compositions may be administered routinely, proactively, such as when at risk, or when the need arises. The compositions may be administered with a meal, before, after, or between meals, or upon notification by an app or electronic device at the onset of the disease or condition the composition is intended to ameliorate.

[0057] (Dose, Content) The dose of the composition of the present invention may be any amount that achieves the desired effect. The dose can be appropriately determined taking into consideration various factors such as the age, weight, and symptoms of the subject.

[0058] The daily dose of the composition can be 0.01 g or more of hesperetin (or the total amount of free hesperetin if multiple active ingredients are used), 0.05 g or more, 0.1 g or more, preferably 0.15 g or more, more preferably 0.2 g or more, even more preferably 0.4 g or more, even more preferably 0.8 g or more, and even more preferably 1 g or more. Studies of changes in bacterial flora due to polyphenol intake have shown changes at around 100 mg / day (Melanie Le Sayec et al. Clinical Nutrition 41 (2022) 2549e2561. https: / / doi.org / 10.1016 / j.clnu.2022.08.024). Therefore, it is believed that such an amount will produce the desired effect. The upper limit of the active ingredient per day, whatever the lower limit, can be 10 g or less, 7 g or less, 5 g or less, 4 g or less, 3 g or less, or 2 g or less.

[0059] Administration may be once a day or multiple times a day, for example, 2 to 10 times. The amount of active ingredient per dose can be 0.003 g or more, 0.015 g or more, or 0.01 g or more, preferably 0.015 g or more, more preferably 0.02 g or more, even more preferably 0.04 g or more, even more preferably 0.08 g or more, and even more preferably 0.1 g or more. The upper limit of the amount of active ingredient per dose, regardless of the lower limit, can be 1 g or less, 0.7 g or less, 0.5 g or less, 0.4 g or less, 0.3 g or less, or 0.2 g or less.

[0060] The content of the active ingredient in the composition can be adjusted appropriately depending on the form of the composition. For example, the content of the active ingredient per solid content of the composition can be 5% or more, preferably 10% or more, more preferably 20% or more, and even more preferably 25% or more. The upper limit of the active ingredient per solid content, regardless of the lower limit, can be 90% or less, 80% or less, 70% or less, 60% or less, or 50% or less. In the present invention, % means % by mass unless otherwise specified.

[0061] When the composition is a liquid, the content of the active ingredient can be 0.01% or more, preferably 0.03% or more, more preferably 0.06% or more, and even more preferably 0.1% or more. When the composition is a liquid, the upper limit of the content of the active ingredient, regardless of the lower limit, can be 10% or less, 8% or less, 6% or less, 4% or less, or 3% or less.

[0062] (Other Ingredients, Additives) In the present invention, the composition may contain other active ingredients or nutritional ingredients that are acceptable as foods or pharmaceuticals. Examples of such ingredients include lipids (e.g., milk fat, vegetable oil, medium-chain fatty acid-containing oil), proteins (e.g., milk protein, milk protein concentrate (MPC), whey protein concentrate (WPC), whey protein isolate (WPI), α-lactalbumin (α-La), β-lactoglobulin (β-Lg), heat-denatured whey protein, and enzyme-treated whey protein), amino acids (e.g., lysine, arginine, glycine, alanine, glutamic acid, leucine, isoleucine, valine), carbohydrates (glucose, sugar, sucrose, fructose, maltose, trehalose, erythritol, maltitol, palatinose, xylitol, dextrin), electrolytes (e.g., sodium, potassium, calcium, magnesium), vitamins (e.g., vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D, vitamin E, vitamin K, biotin, folic acid, pantothenic acid, and nicotinic acids), minerals (e.g., copper, zinc, iron, cobalt, manganese), antibiotics, dietary fiber, etc.

[0063] In the present invention, the composition may contain a prebiotic other than the active ingredient. Prebiotics can be defined as indigestible food ingredients that have a beneficial effect on the host and improve the host's health by selectively altering the growth and activity of specific bacteria in the large intestine. The composition may contain one or more prebiotics other than the active ingredient.

[0064] The prebiotics other than the active ingredient are not particularly limited as long as they do not interfere with the effects of the active ingredient contained in the composition. Examples of prebiotics other than the active ingredient include galactooligosaccharides, fructooligosaccharides, xylooligosaccharides, isomaltooligosaccharides, raffinose, lactulose, lactosucrose, soybean oligosaccharides, coffee oligosaccharides, dietary fiber, and gluconic acid.

[0065] The composition may further contain additives acceptable for use as food or pharmaceuticals, such as inert carriers (solid or liquid carriers), excipients, surfactants, binders, disintegrants, lubricants, solubilizers, suspending agents, coating agents, colorants, preservatives, buffers, pH adjusters, emulsifiers, stabilizers, sweeteners, antioxidants, flavors, acidulants, and natural products. More specifically, examples of the additives include water, other aqueous solvents, pharmaceutically acceptable organic solvents, collagen, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymers, sodium alginate, water-soluble dextran, water-soluble dextrin, sodium carboxymethyl starch, pectin, xanthan gum, gum arabic, casein, gelatin, agar, glycerin, propylene glycol, polyethylene glycol, petrolatum, paraffin, stearyl alcohol, stearic acid, human serum albumin, mannitol, sorbitol, lactose, sucralose, stevia, aspartame, acesulfame potassium, citric acid, lactic acid, malic acid, tartaric acid, phosphoric acid, acetic acid, fruit juice, and vegetable juice.

[0066] (Dosage Form / Form) The food composition may be prepared in any form, such as a solid, liquid, mixture, suspension, powder, granules, paste, jelly, gel, or capsule. The food composition of the present invention can also be prepared in any form, such as dairy products, supplements, confectioneries, beverages, energy drinks, seasonings, processed foods, prepared dishes, or soups. More specifically, the composition of the present invention can be in the form of liquid food, semi-liquid food, jelly, gel, powder, infant formula, infant formula, liquid milk, powdered or liquid milk for pregnant or nursing women, fermented milk, bar, mousse, chocolate, biscuits, ice cream, fermented milk, lactic acid bacteria beverage, dairy beverage, soft drink, tablet, cheese, bread, biscuit, cracker, pizza crust, food for the sick, nutritional food, frozen food, or processed food. It can also be in the form of granules, powder, paste, or concentrated liquid for administration by mixing with beverages or foods. The granules and powders can be cube-shaped or stick-shaped (single-serving amounts packaged). In the present invention, modified powdered milk refers to raw milk, cow's milk, special cow's milk, or raw buffalo milk, or foods made from these as raw ingredients, processed or used as the main ingredient, to which nutrients necessary for infants have been added, and made into a powder, as defined in the Ministerial Ordinance on Milk, etc. In the present invention, modified liquid milk refers to raw milk, cow's milk, special cow's milk, or raw buffalo milk, or foods made from these as raw ingredients, processed or used as the main ingredient, to which nutrients necessary for infants have been added, and made into a liquid, as defined in the Ministerial Ordinance on Milk, etc.

[0067] Formulated powdered milk or formulated liquid milk (these are sometimes collectively referred to as formulae. In addition, formulae are sometimes explained using formulae as an example, but the explanation also applies to formulae liquid milk) can be for infants, follow-up (to supplement weaning food), low birth weight infants, children, adults, pregnant women, nursing women, the elderly, those with allergies, those with lactose intolerance, and those with inborn errors of metabolism. Preferred examples of formulae are those for infants, follow-up, low birth weight infants, and children.

[0068] The enteric formulation and the pharmaceutical composition can be in any dosage form suitable for oral administration, such as a solid formulation such as a tablet, granules, powder, pill, or capsule; a liquid formulation such as a solution, suspension, or syrup; a gel; or an aerosol.

[0069] When the composition is intended to control bacteria belonging to the genus Fusobacterium in the oral cavity, it is preferably in a form that can be retained in the oral cavity for a relatively long period of time. Examples of such forms include mouthwash, candy, gummies, chewing gum, lozenges, sublingual tablets, oral patches, ointments, pastes, jellies, and oral tablets. When the composition is intended for oral use, it may be a toothpaste composition or a mouthwash composition.

[0070] (Other) In the production of the composition of the present invention, the stage of blending the active ingredient can be selected as appropriate. The stage of blending is not particularly limited as long as it does not significantly impair the properties of the active ingredient. For example, the active ingredient can be blended by mixing it with raw materials. Alternatively, the active ingredient can be added at the final stage of production to produce a composition containing the active ingredient.

[0071] The compositions of the present invention may be labeled with the intended use (application) and may also be labeled to recommend administration to a particular subject.

[0072] The composition of the present invention can be labeled with its intended use (application), and in one embodiment, it is recommended for administration to a specific subject. Examples of subjects to be labeled are as described above. In one embodiment, the composition of the present invention can be labeled with the function of the composition or active ingredient or a usage method based on that function. For example, it can be labeled that the composition can control the growth of Fusobacterium bacteria in the intestinal tract, can inhibit the growth of Fusobacterium bacteria in the intestinal tract, can treat diseases or conditions that are improved by controlling (preferably inhibiting) the growth of Fusobacterium bacteria in the intestinal tract, can be used as a prebiotic, can be used as a synbiotic (a combination of probiotics and prebiotics), or can treat inflammatory bowel disease and colorectal cancer. Note that a period such as "temporary" or "long-term" may be added to the beginning of each statement, as appropriate.

[0073] Representation can be explicit or implicit. Examples of explicit representations are direct descriptions on tangible objects such as the product itself, packaging, containers, labels, tags, etc., while examples of implicit representations (which can also be called implied) include advertising and promotional activities by place or means such as websites, stores, pamphlets, exhibitions, seminars such as media seminars, books, newspapers, magazines, television, radio, mail, email, and audio.

[0074] In one embodiment, the recommendation to consume the composition is displayed personally. Such a display can be made using a document (whether written or electronic) addressed to the subject, the subject's tablet device, smartphone, personal computer, the subject's social media, etc. Furthermore, such a display can be made together with the display of the results of any test or analysis, such as a gut flora test, a fecal substance test, fecal metabolome analysis, an oral flora test, a substance test in the oral cavity (e.g., saliva, tongue coating, dental plaque), or a metabolome analysis selected from saliva, tongue coating, and dental plaque.

[0075]

[0013] In one aspect, there is provided a method for controlling bacteria belonging to the genus Fusobacterium in the intestinal tract of a subject, or a method for reducing the risk of at least one of inflammatory bowel disease and colorectal cancer in a subject, the method comprising the step of having the subject ingest a composition or a food material comprising any one selected from the group consisting of flavonoids and transglycosylation products thereof that release hesperetin in the intestine, and hesperetin, based on the presence or amount of bacteria belonging to the genus Fusobacterium in the subject's intestinal tract. In another aspect, there is provided a method for suggesting to a subject to consume a composition, a method for supporting a subject's diet, or a method for supporting a subject's health, the method comprising the step of suggesting to a subject to consume a composition or a food material comprising any one selected from the group consisting of flavonoids and transglycosylation products thereof that release hesperetin in the intestine, and hesperetin, based on the presence or amount of bacteria belonging to the genus Fusobacterium in the subject's intestinal tract. These methods may include other steps, for example, any of the following: collecting a fecal sample from a subject or having the subject collect a sample; analyzing the sample for the presence or amount of Fusobacterium bacteria; displaying the analysis results; determining, based on the analysis results, whether it is desirable or necessary for the subject to reduce the number of Fusobacterium bacteria; displaying the determination results; and displaying a composition or food ingredient product containing any one selected from the group consisting of flavonoids and their glycosyltransferases that liberate hesperetin in the intestine, and hesperetin.

[0076]

[0013] In one aspect, there is provided a method for controlling bacteria belonging to the genus Fusobacterium in the oral cavity of a subject, or a method for reducing the risk of any of periodontal disease, halitosis, oral cancer, esophageal cancer, aspiration pneumonia, and Alzheimer's disease in a subject, the method comprising the step of having the subject ingest a composition or food material containing any one selected from the group consisting of a flavonoid and a transglycosylation product thereof that releases hesperetin in the oral cavity, and hesperetin, or gargling with the composition, based on the presence or amount of bacteria belonging to the genus Fusobacterium in the oral cavity of the subject. In another aspect, there is provided a method for suggesting to a subject to ingest a composition, a method for suggesting to a subject to gargle with the composition, a method for supporting a subject's dietary habits, or a method for supporting a subject's health, the method comprising the step of suggesting to ingest a composition or food material containing any one selected from the group consisting of a flavonoid and a transglycosylation product thereof that releases hesperetin in the oral cavity, and hesperetin, or suggesting to gargle with the composition, based on the presence or amount of bacteria belonging to the genus Fusobacterium in the oral cavity of the subject. These methods may include other steps, for example, any of the following: collecting a sample from the oral cavity of a subject, or having the subject collect a sample; analyzing the sample for the presence or amount of Fusobacterium bacteria; displaying the analysis results; determining, based on the analysis results, whether it is desirable or necessary for the subject to reduce the number of Fusobacterium bacteria; displaying the determination results; and displaying a composition or food ingredient product containing any one selected from the group consisting of flavonoids and their glycosyltransferases that release hesperetin in the oral cavity, and hesperetin.

[0077] The present invention will be described in more detail below using examples, although the technical scope of the present invention is not limited to these examples.

[0078] Example 1: Feces were cultured in a medium containing hesperidin (Fujifilm Wako Pure Chemical Industries, Ltd.), and the number of Fusobacterium bacteria was quantified to confirm that hesperidin reduces the number of Fusobacterium bacteria. Specifically, feces from six individuals were cultured in mYCFA medium containing 0.3% (w / v) hesperidin (0.15% (w / v) as hesperetin) at 37°C under anaerobic conditions for 16 hours, and DNA was extracted from each culture using a Maxwell® RSC Instrument (Promega Corporation).

[0079] The bacterial count was measured as follows. Quantitative PCR was performed on DNA extracted from the culture medium in which the number of Fusobacterium bacteria had been measured in advance, and a standard curve was created. Quantitative PCR was performed on the DNA obtained above, and the number of Fusobacterium bacteria was calculated based on the standard curve. The results were then corrected to the number of bacteria per mL of culture medium. The primer sequences used for quantitative PCR are shown in Table 1. DNA extracted from Fusobacterium nucleatum subsp. nucleatum JCM 8532T (the type species of Fusobacterium bacteria) was used as the standard.

[0080]

[0081] For stool samples in which Fusobacterium is not detected before the culture stage, 1 × 10 Fusobacterium nucleatum JCM8532T was added at the start of the culture. 4 cfu / ml was added.

[0082] As a result, the number of Fusobacterium bacteria was reduced in all cultures to which hesperidin was added compared to the control (Purewater was added instead of hesperidin) (Figure 1).

[0083] These results suggest that hesperidin may reduce the number of Fusobacterium bacteria.

[0084] Example 2: Next, Fusobacterium was cultured on an agar medium containing hesperetin (Fujifilm Wako Pure Chemical Industries, Ltd.), and it was confirmed that Fusobacterium did not grow in the medium containing hesperetin. Specifically, Fusobacterium nucleatum JCM JCM8532T was inoculated onto GAM agar medium containing 0.3% (w / v) hesperetin and onto GAM agar medium not containing hesperetin, and cultured at 37°C under anaerobic conditions for 48 hours, after which the number of colonies was counted. As a result, colonies were detected in the medium without hesperetin, but no colonies appeared in the medium containing hesperetin (Figure 2).

[0085] These results indicate that hesperetin inhibits the growth of Fusobacterium.

[0086] [Summary] As described above, flavonoids such as hesperidin, which release hesperetin by β-glucosidase, and their transglycosylation products, as well as hesperetin, can be used to reduce the number of Fusobacterium bacteria in the digestive tract (oral cavity and intestinal tract) where β-glucosidase-producing bacteria are present. Flavonoids and their transglycosylation products, which release hesperetin by β-glucosidase produced by enteric bacteria in the oral cavity and intestine, as well as hesperetin, can be used to reduce the number of Fusobacterium bacteria in the oral cavity and intestinal flora. Furthermore, hesperetin itself can be used to reduce the number of Fusobacterium bacteria. Fusobacterium bacteria have recently been suggested to be involved in colon cancer (e.g., Non-Patent Document 1). F. nucleatum has been isolated from gastrointestinal infections such as appendicitis (Non-Patent Document 2), and its association with inflammatory bowel disease has also been suggested (Non-Patent Documents 3, 4, and 5). Oral Fusobacterium bacteria are known to cause periodontal disease, halitosis, oral cancer, esophageal cancer, aspiration pneumonia, and Alzheimer's disease (Non-Patent Documents 6, 7, 8, 9, 10). Therefore, flavonoids and their glycosyltransferases that release hesperetin in the digestive tract by β-glucosidase, as well as hesperetin, can be used to treat these diseases.

[0087] [Production Example 1: Fermented Milk] A yogurt base mix was prepared by mixing milk, a dairy product (derived from milk), and water so that the final product would have a non-fat milk solids content of 9.5%, a milk fat content of 3.0%, and a monoglucosyl hesperidin content of 0.2%. The prepared yogurt base mix was homogenized, sterilized by heating at 95°C for 5 minutes, and then cooled to approximately 40°C. Lactobacillaceae lactic acid bacteria were inoculated into this sterilized yogurt base mix, and the Lactobacillaceae lactic acid bacteria were inoculated at a concentration of 5 x 10 per 100g. 9 The yogurt is fermented until the number of Fusobacterium species reaches 100 or more, thereby producing yogurt for controlling Fusobacterium species in the intestinal flora.

[0088] [Production Example 2: Health Food] Each ingredient (acidulant, sweetener, stabilizer, fruit juice, flavoring, monoglycosyl hesperidin, and water) was mixed to obtain a soft drink, which was then bottled in approximately 500 mL portions to prepare a drink for controlling Fusobacterium spp. Monoglycosyl hesperidin was blended at 400 mg per 500 mL.

[0089] [Production Example 3: Tablets] The raw materials (excipient, acidulant, sweetener, stabilizer, fruit juice, flavoring, and monoglycosyl hesperidin) are mixed and compressed into tablets to prepare tablets for controlling Fusobacterium in the oral cavity. 6 mg of monoglycosyl hesperidin is added per 1 g of tablet.

[0090] [Production Example 4: Gum] The ingredients (maltitol, gum base, acidulant, sweetener, thickener (gum arabic), hesperidin, glazing agent, and coloring agents (gardenia, safflower yellow)) were mixed to produce a granular gum for controlling Fusobacterium in the oral cavity. Monoglycosyl hesperidin was added at 4 mg per 1 g of gum.

[0091] [Production Example 5: Capsules] Hayashibara Hesperidin (registered trademark) S (transglycosylated hesperidin) is powder-filled into enteric-coated capsules in amounts of 20 mg each to produce a pharmaceutical for controlling Fusobacterium bacteria in the intestines, for intestinal disorders, for regulating bowel movements, or for improving loose stools, constipation, and abdominal bloating.

[0092] [Production Example 6: Enteric-coated supplement (tablets)] A powder mixture of raw materials containing hesperetin is tableted and coated to obtain tablets containing 10 to 40 mg of hesperetin per tablet. The additives other than Lactobacillaceae lactic acid bacteria are corn starch, dextrin, lactose hydrate, precipitated calcium carbonate, candy powder, white sugar, talc, and magnesium stearate.

[0093] Preparation Example 7: Enteric-Release Supplement (Capsules) Hesperetin is filled at 10-40 mg per capsule into pulse capsules (Pulsincap®, Scherer DDS, Ltd.), which are designed to release their contents after approximately 8 hours of contact with the contents of the gastrointestinal tract.

[0094] The present invention supports the maintenance and improvement of people's health by providing a composition for controlling Fusobacterium bacteria. The present invention also provides a food composition and a method for producing food that support the maintenance and improvement of people's health. Furthermore, the present invention can improve the nutrition of various people, ensure healthy lifestyles, and promote welfare.

[0095] [Sequences listed in the sequence listing] SEQ ID NO: 1 PCR Forward Primer SEQ ID NO: 2 PCR Reverse Primer

Claims

1. A composition for controlling bacteria belonging to the genus Fusobacterium in the digestive tract, comprising any one selected from the group consisting of flavonoids and their glycosyltransferases that liberate a compound represented by formula I in the digestive tract, and the compound represented by formula I. (wherein R1 to R4 are the following combinations):

2. A composition for controlling bacteria belonging to the genus Fusobacterium, comprising any one selected from the group consisting of flavonoids and their glycosyltransferases which release a compound represented by formula I upon hydrolysis by glucosidase, and the compound represented by formula I. (wherein R1 to R4 are the following combinations):

3. The composition according to claim 1 or 2, wherein R1 to R4 are the following combinations:

4. The composition according to claim 1 or 2, wherein the compound represented by formula I is hesperetin, the flavonoid that liberates hesperetin is hesperidin, and the composition comprises any one selected from the group consisting of hesperidin, a glycosyltransferase of hesperidin, and hesperetin.

5. The composition according to claim 4, wherein the glycosyltransferase of hesperidin is glucosylhesperidin.

6. A composition for controlling bacteria belonging to the genus Fusobacterium, comprising hesperetin.

7. The composition according to claim 1, 2, or 6, wherein the control of bacteria belonging to the genus Fusobacterium is a reduction in the number of bacteria belonging to the genus Fusobacterium.

8. The composition of claim 1, 2 or 6 for use as any one selected from the group consisting of prebiotics and synbiotics.

9. The composition described in claim 1, 2, or 6 for treating any one selected from inflammatory bowel disease, colon cancer, periodontal disease, halitosis, oral cancer, esophageal cancer, aspiration pneumonia, and Alzheimer's disease in a subject in whom Fusobacterium genus has been detected in at least one of the intestinal tract and feces.

10. The composition of claim 1, 2, or 6 for treating any one selected from inflammatory bowel disease, colon cancer, periodontal disease, halitosis, oral cancer, esophageal cancer, aspiration pneumonia, and Alzheimer's dementia.

11. The composition according to claim 1, comprising as an active ingredient any one selected from the group consisting of hesperetin and glycosyltransferases of hesperidin, and the digestive tract is the oral cavity.

12. The composition of claim 1, wherein the digestive tract is the intestine.

13. A food composition comprising any one selected from the group consisting of flavonoids and their glycosyltransferases that liberate a compound represented by formula I in the digestive tract, as defined in claim 1, and compounds represented by formula I (excluding glycosyltransferase hesperidin), which is any one selected from the group consisting of functional foods, foods for specified health uses, fermented milk, milk beverages, and powdered foods.

14. A pharmaceutical composition comprising any one selected from the group consisting of a flavonoid and its glycosyltransferase which liberates a compound represented by formula I in the digestive tract, and the compound represented by formula I, as defined in claim 1.

15. An enteric formulation comprising any one selected from the group consisting of a flavonoid and its glycosyltransferase which liberates a compound represented by formula I in the digestive tract, and the compound represented by formula I, as defined in claim 1.

16. A composition according to claim 13 or 14, or a formulation according to claim 15, containing 0.01 g or more and 7 g or less of any one selected from the flavonoids and their glycosyltransferases, and the compounds.