Fusobacterium controlling composition and use thereof

Abstract

The present invention addresses the problem of controlling Fusobacterium with a method that is considered to have little influence on beneficial bacteria. Provided is a Fusobacterium controlling composition comprising water soluble dietary fiber. The water soluble dietary fiber is preferably isomaltodextrin.

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, and 5).

[0003] On the other hand, isomaltodextrin is a type of water-soluble dietary fiber. Water-soluble dietary fiber or isomaltodextrin, sometimes in combination with other dietary fibers, is known to have several effects related to the digestive tract and defecation. For example, Patent Document 1 describes an agent for improving maximum gastrointestinal transit time, characterized by containing one or more of indigestible sugars, low-viscosity water-soluble dietary fiber, and magnesium compound. Furthermore, Patent Document 2 describes a composition for promoting the growth of Fusicatenibacter bacteria, which contains water-soluble dietary fiber as an active ingredient, and lists isomaltodextrin as an example of water-soluble dietary fiber. Furthermore, Patent Document 3 describes a composition for preventing, treating, or suppressing intestinal disorders, which is characterized by containing three or more types of indigestible carbohydrate raw materials selected from oligosaccharides (excluding chitin oligosaccharides) and / or dietary fibers, and lists preferred examples of indigestible dietary fibers as inulin, guar gum hydrolysate, psyllium seed gum, indigestible dextrin, isomaltodextrin, cellulose, β-glucan, glucomannan, and agarose.

[0004] It has been reported that the anti-inflammatory effect of isomaltodextrin on dextran sulfate sodium (DSS)-induced colitis was evaluated in a mouse model (Non-Patent Document 6). This report stated that although isomaltodextrin was unable to prevent DSS-induced weight loss or colon shortening, it tended to reduce the inflammatory cytokines TNF-α and IL-6, suppress the expression of IL-1β, MCP-1, and IL-17, and increase the anti-inflammatory cytokine IL-10. It also stated that the anti-inflammatory effect of isomaltodextrin was mediated by suppressing the expression of Toll-like receptor-4.

[0005] Furthermore, Patent Document 4 describes a preparation for dieting and / or improving the intestinal flora, characterized by encapsulating Bifidobacterium breve, Bifidobacterium longum, and N-acetylglucosamine in an acid-resistant hard capsule, and describes that in the group administered with the preparation in the examples, a significant increase in Lachnospiraceae sp. and Roseburia sp. and a significant decrease in Fusobacterium sp., Prevotella sp., and Enterobacteriaceae sp. are confirmed. Patent Document 5 describes a composition containing Bifidobacterium bacteria as an active ingredient for inhibiting the adsorption of Fusobacterium bacteria to mucosal cells, and a pharmaceutical composition containing Bifidobacterium bacteria as an active ingredient for preventing or treating a disease or symptom caused by inflammation of mucosal cells and selected from the group consisting of ulcerative colitis, colon cancer, esophageal cancer, premature birth, and periodontal disease. Non-Patent Document 7 describes that when subjects ingested 15 g / day of oligofructose for 15 days, the number of bacteroides, clostridia, and fusobacteria decreased. Patent Document 6 describes a dietary fiber preparation containing a first soluble fiber and a second soluble fiber, which exhibits greater prebiotic activity than the individual prebiotic activities of the first soluble fiber and the second soluble fiber, and cites fructooligosaccharide (FOS) as an example of the second soluble fiber. This document also describes that FOS can enhance positive immune responses by promoting the growth of bifidobacteria and suppress the growth of pathogenic bacteria that may cause diarrhea, thereby promoting intestinal health.

[0006] International Publication WO2006 / 054710 JP 2020-83830 A JP 2020-178685 A JP 2016-166169 A International Publication No. 2019 / 117212 (Patent Publication No. 2020-178685) JP 2012-111771 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]K. Majumder et. al. Intervention of Isomaltodextrin Mitigates Intestinal Inflammation in a Dextran Sodium Sulfate-Induced Mouse Model of Colitis via Inhibition of Toll-like Receptor-4. J Agric Food Chem. 2017;65(4):810-817. doi: 10.1021 / acs.jafc.6b04903.Glenn R. Gibson et. al. Selective Stimulation of Bifidobacteria in the Human Colon by Oligofructose and Inulin. Gastroenterology 1995;108:975-982.

[0008] There are no known effective materials that can control Fusobacterium species.

[0009] Because Fusobacterium bacteria live in a complex symbiotic relationship with other intestinal bacteria in the intestine, selective control of Fusobacterium bacteria is thought to be difficult. For example, the administration of antibiotics is thought to reduce not only Fusobacterium bacteria but also beneficial bacteria. It is desirable to control Fusobacterium bacteria using a method that is thought to have minimal impact on beneficial bacteria.

[0010] The present invention provides the following: [1] A composition for controlling bacteria belonging to the genus Fusobacterium, comprising water-soluble dietary fiber. [2] The composition according to 1, wherein the water-soluble dietary fiber is a low-viscosity water-soluble dietary fiber. [3] The composition according to 1 or 2, wherein the water-soluble dietary fiber is a hyperbranched α-glucan. [4] The composition according to any one of 1 to 3, wherein the water-soluble dietary fiber is isomaltodextrin. [5] The composition according to any one of 1 to 4, wherein the control of bacteria belonging to the genus Fusobacterium is a reduction in the number of bacteria belonging to the genus Fusobacterium. [6] The composition according to any one of 1 to 5, for use as a prebiotic or synbiotic. [7] The composition according to any one of 1 to 6, for treating at least one of inflammatory bowel disease and colorectal cancer via control of bacteria belonging to the genus Fusobacterium. [8] A composition for treating colorectal cancer, comprising water-soluble dietary fiber, preferably an α-glucan, more preferably a hyperbranched α-glucan. [9] The composition according to 8, wherein the multi-branched α-glucan is isomaltodextrin.

[10] The composition according to 8 or 9, for administration to a subject having bacteria belonging to the genus Fusobacterium in the intestine.

[11] A pharmaceutical composition comprising isomaltodextrin.

[12] The pharmaceutical composition according to 13, for use in an intestinal disease.

[13] An enteric-coated formulation comprising isomaltodextrin.

[0011]

[14] 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 containing water-soluble dietary fiber based on the presence or absence or amount of bacteria belonging to the genus Fusobacterium in the intestine of the subject.

[15] A method for suggesting intake of a composition to a subject, a method for supporting the dietary habits of a subject, or a method for supporting the health of a subject, comprising the following steps: suggesting intake of a composition containing water-soluble dietary fiber based on the presence or absence or amount of bacteria belonging to the genus Fusobacterium in the intestine of the subject.

[0012]

[21] A composition comprising water-soluble dietary fiber for use in a method for controlling bacteria belonging to the genus Fusobacterium. Use of water-soluble dietary fiber 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 a composition comprising water-soluble dietary fiber to a subject. Use or non-therapeutic use of a composition comprising water-soluble dietary fiber for controlling bacteria belonging to the genus Fusobacterium.

[22] The composition, use in manufacture, method or non-therapeutic method, or use or non-therapeutic use according to 21, wherein the water-soluble dietary fiber is a low-viscosity water-soluble dietary fiber.

[23] The composition, use in manufacture, method or non-therapeutic method, or use or non-therapeutic use according to 21 or 22, wherein the water-soluble dietary fiber is a multibranched α-glucan.

[24] The composition, use in manufacture, method or non-therapeutic method, or use or non-therapeutic use according to any one of 21 to 23, wherein the water-soluble dietary fiber is isomaltodextrin.

[25] The composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to any one of items 21 to 24, wherein the control of bacteria belonging to the genus Fusobacterium is a reduction in the number of bacteria belonging to the genus Fusobacterium.

[26] The composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to any one of items 21 to 25, wherein the composition is for use as a prebiotic or synbiotic.

[27] The composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to any one of items 21 to 26, for treating at least one of inflammatory bowel disease and colorectal cancer via control of bacteria belonging to the genus Fusobacterium.

[28] A composition comprising water-soluble dietary fiber, preferably α-glucan, more preferably hyperbranched α-glucan, for use in a method for treating colorectal cancer. Use of water-soluble dietary fiber, preferably α-glucan, more preferably hyperbranched α-glucan, in the production of a composition for treating colorectal cancer.A method or non-therapeutic method for treating colorectal cancer, comprising the step of administering to a subject a composition comprising water-soluble dietary fiber, preferably α-glucan, more preferably hyperbranched α-glucan. Use or non-therapeutic use of a composition comprising water-soluble dietary fiber, preferably α-glucan, more preferably hyperbranched α-glucan, for treating colorectal cancer.

[29] The composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to 28, wherein the hyperbranched α-glucan is isomaltodextrin.

[30] The composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to 28 or 29, wherein the composition is for administration to a subject having bacteria belonging to the genus Fusobacterium in their intestines.

[31] A pharmaceutical composition comprising isomaltodextrin.

[32] The pharmaceutical composition according to 11, for use in an intestinal disease.

[33] An enteric-coated formulation comprising isomaltodextrin.

[0013] The use of isomaltodextrin as a prebiotic provides a means to reduce Fusobacterium, which in turn may prevent or improve colon cancer and inflammatory bowel disease.

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

[0015] The present invention relates to a composition for controlling bacteria of the genus Fusobacterium, which contains water-soluble dietary fiber as an active ingredient.

[0016] [Active Ingredient] The composition of the present invention contains water-soluble dietary fiber as an active ingredient. In the present invention, dietary fiber is defined as the total amount of indigestible components in food that are not digested by digestive enzymes. Proteins, lipids, and carbohydrates are broken down (digested) by digestive enzymes in the digestive tract and absorbed into the body from the small intestine, but dietary fiber is a component that passes through the small intestine without being affected by digestive enzymes and reaches the large intestine. Dietary fiber includes starch (resistant dextrin) and oligosaccharides, which are resistant to digestion. Dietary fiber can be broadly classified into water-soluble and water-insoluble dietary fibers. The composition of the present invention contains water-soluble dietary fiber. The composition may contain only one type of water-soluble dietary fiber or two or more types as an active ingredient. Water-soluble dietary fiber has been reported to be easily utilized by intestinal bacteria. Water-soluble dietary fiber also has many advantages, such as being easily dispersed in the system during production and preventing residue on the production line and adhesion to filters. "Containing an active ingredient" means that the composition contains an effective amount of the ingredient to achieve the intended function, and that the ingredient is specified in the label as contributing to the intended purpose. In foods with functional claims, active ingredients are sometimes called functional ingredients (ingredients that contribute to specific health purposes (excluding those related to reducing disease risk)).

[0017] Examples of water-soluble dietary fiber include isomaltodextrin, barley β-glucan, polydextrose, indigestible dextrin, indigestible glucan, alginic acid, fucoidan, guar gum hydrolyzate, inulin, and acacia dietary fiber.

[0018] In one embodiment, the composition contains, as an active ingredient, a low-viscosity water-soluble dietary fiber. The low-viscosity water-soluble dietary fiber refers to a water-soluble dietary fiber that has a low viscosity in an aqueous solution. Specifically, the low-viscosity water-soluble dietary fiber refers to a water-soluble dietary fiber whose viscosity, measured in a 1% aqueous solution at 20°C using a Brookfield viscometer, is 100 mPa·s or less.

[0019] In one embodiment, the composition contains, as an active ingredient, a highly branched α-glucan, a low-viscosity water-soluble dietary fiber, such as isomaltodextrin.

[0020] Isomaltodextrin is a water-soluble dietary fiber produced by the action of enzymes on starch. Compared to regular dextrin obtained by hydrolyzing starch, it has a higher proportion of branched structures with α-1,6 glucoside bonds and does not contain glucose with β-glucosidic bonds. Isomaltodextrin is composed only of glucose with α-bonds and has more branches than the starch it is made from, so it is considered a type of hyperbranched α-glucan.

[0021] In a preferred embodiment, isomaltodextrin having a DE of 10 or less is used. A DE of 9 or less is more preferable, a DE of 8 or less is even more preferable, and a DE of 7.0 to 7.5 is even more preferable. DE here means dextrose equivalent, and is a value that represents a relative measure when the reducing power of dextrose (glucose) is set to 100. Isomaltodextrin has a lower viscosity than maltodextrin with a similar DE.

[0022] The weight-average molecular weight of the isomaltodextrin used is preferably 500 to 10,000, more preferably 3,000 to 7,000, and even more preferably 4,500 to 5,500. The number-average molecular weight of the isomaltodextrin is preferably 1,000 to 5,000, and more preferably 2,000 to 3,000.

[0023] [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 intestinal flora. In a more preferred embodiment, the composition can be used to control Fusobacterium bacteria in human intestinal flora. Controlling Fusobacterium bacteria in intestinal flora refers to controlling bacteria belonging to the genus Fusobacterium in a state where various bacteria living in the intestine are present. Controlling bacteria refers to controlling the number of bacteria, regulating intestinal flora including Fusobacterium bacteria, and assisting in regulating intestinal flora including Fusobacterium bacteria. In a preferred embodiment, the composition is used to reduce the number of Fusobacterium bacteria or assist in reducing the number of Fusobacterium bacteria.

[0024] The intestine refers to the digestive organ in humans and animals where bacteria normally reside and where ingested food is digested and absorbed. The intestine includes the small intestine and the large intestine, and preferably the large intestine.

[0025] 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

[0026] 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).

[0027] 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, incubated for a certain period of time. After that, the component to be evaluated is added and incubated under appropriate conditions (e.g., incubation at 37°C under anaerobic conditions similar to those in the intestine for 16 hours). If the feces does not contain Fusobacterium bacteria, any available Fusobacterium bacteria is added at the start of incubation and then incubated. 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.

[0028] 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. TMFor 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.

[0029] In general, the intestinal flora contained in the bacterial flora may include, but is not limited to, the following bacteria. In one embodiment, the composition can control the number of bacteria of the genus Fusobacterium while maintaining the numbers of bacteria of the genus Parabacteroides and Bifidobacterium. In another embodiment, the composition controls the number of bacteria of the genus Fusobacterium while not affecting any of the following bacteria, or has only a small effect, if any. Bacteroides uniformis, Alistipes putredinis, Parabacteroides merdae, Dorea longicatena, Ruminococcus bromii L2-63, Bacteroides caccae, Clostridium sp SS2-1, Bacteroides thetaiotaomicron VPI-5482, Eubacterium hallii, Ruminococcus torques L2-14, unknown sp SS3 4, Ruminococcus sp SR1 5, Faecalibacterium prausnitzii SL3 3, Ruminococcus lactaris, Collinsella aerofaciens, Dorea formicigenerans, Bacteroides vulgatus ATCC 8482, Roseburia intestinalis M50 1, Bacteroides sp. 2_1_7, Eubacterium siraeum 70 3, Parabacteroides distasonis ATCC 8503, Bacteroides sp. 9_1_42FAA, Bacteroides ovatus, Bacteroides sp. 4_3_47FAA, Bacteroides sp. 2_2_4, Eubacterium rectale M104 1, Bacteroides xylanisolvens XB1A, Coprococcus comes SL7 1, Bacteroides sp. D1, Bacteroides sp.D4, Eubacterium ventriosum, Bacteroides dorei, Ruminococcus obeum A2-162, Subdoligranulum variabile, Bacteroides capillosus, Streptococcus thermophilus LMD-9, Clostridium leptum, Holdemania filiformis, Bacteroides stercoris, Coprococcus eutactus, Clostridium sp M62 1, Bacteroides eggerthii, Butyrivibrio crossotus, Bacteroides finegoldii, Parabacteroides johnsonii, Clostridium sp L2-50, Clostridium nexile, Bacteroides pectinophilus, Anaerotruncus colihominis, Ruminococcus gnavus, Bacteroides intestinalis, Bacteroides fragilis 3_1_12, Clostridium asparagiforme, Enterococcus faecalis TX0104, Clostridium scindens, Blautia hansenii (Nature 464, 59-65 (2010) doi:10.1038 / nature08821).

[0030] The Fusobacterium bacteria of the present invention are bacteria present in the human oral cavity and intestines 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 selected from the group consisting of inflammatory bowel disease and colon cancer.

[0031] Inflammatory bowel diseases include ulcerative colitis and Crohn's disease.

[0032] When used to treat inflammatory bowel disease, the composition may be configured to exclude any of the following: oligosaccharides (e.g., raffinose, stachyose, galactooligosaccharides, isomaltooligosaccharides, lactulose, xylooligosaccharides, agarooligosaccharides, mannooligosaccharides, or fructooligosaccharides), inulin, pectin, modified pectin, guar gum, hydrolyzed guar gum, psyllium seed gum, karaya gum, tragacanth gum, gum arabic, resistant starch, resistant dextrin, polydextrose, cellulose, hemicellulose, soybean polysaccharides, β-glucan, glucomannan, galactomannan, chondroitin sulfate, hyaluronic acid, levan, lignin, alginic acid and its salts, agarose, and chitosan. Fructooligosaccharides are also called short-chain fructose polymers or oligofructose (Patent Document 6, Non-Patent Document 7).

[0033] When used to treat inflammatory bowel disease, in a particularly preferred embodiment, the composition contains isomaltodextrin as an active ingredient and may contain no other dietary fiber.

[0034] When used for the treatment of colon cancer, the composition may contain various water-soluble dietary fibers as active ingredients, but in a particularly preferred embodiment, the composition contains isomaltodextrin as an active ingredient and may not contain any other dietary fibers.

[0035] The composition can also be used to treat a variety of diseases or conditions that can be improved by controlling the growth of Fusobacterium bacteria in the intestine.

[0036] 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).

[0037] (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 intestine, particularly to reduce the number of bacteria; subjects with any one selected from the group consisting of inflammatory bowel disease and colorectal cancer; and subjects with a disease or condition that can be improved by controlling the proliferation of Fusobacterium bacteria in the intestine. 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 food or the like. "Administer" can be read as "intake" (or "ingestion"), and "administer" can be read as "administered".

[0038] Subjects for whom it is desirable or necessary to control Fusobacterium bacteria in their intestines include subjects in whom Fusobacterium bacteria have been detected in at least one of their intestines and feces, and subjects in whom relatively high numbers of Fusobacterium bacteria have been detected in at least one of their intestines and feces. Furthermore, subjects for whom it is desirable or necessary to control Fusobacterium bacteria in their intestines 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. The subject also includes a subject for whom it is desired to reduce the number of Fusobacterium bacteria in at least one of the intestines and feces, a subject for whom it is desired to reduce the risk of colorectal cancer, and a subject for whom it is desired to reduce the risk of inflammatory bowel disease. In one embodiment, the subject is a subject for whom Fusobacterium bacteria have been detected in at least one of the intestines and feces, or a subject for whom it has been recognized that the number of Fusobacterium bacteria is relatively high. For example, the number of Fusobacterium bacteria detected in at least one of the intestines and feces, or a relatively high number of Fusobacterium bacteria, means that when a culture solution containing stool collected from a subject at a concentration of 0.1% (w / v) is cultured at 37°C under anaerobic conditions for 16 hours, the number of Fusobacterium bacteria in the culture solution 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 4 cells / g or more, preferably 5×10 4 cells / g or more, preferably 1×10 5 cells / g or more, more preferably 5×10 5This refers to the case where the number of bacteria is more than 10 ...

[0039] In one embodiment, the subject is healthy (not having a medically diagnosed illness). The subject may be a human or a non-human animal.

[0040] The age of the subject 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 to 6 years of age); a child (7 years or older, but less 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, for example, a person 60 years or older, a person 65 years or older, a person 70 years or older, or a person 75 years or older. It has often been reported that the number of Fusobacterium bacteria in the intestine increases with age (e.g., Odamaki et al. BMC Microbiology (2016) 16:90 DOI 10.1186 / s12866-016-0708-5).

[0041] [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.

[0042] (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.

[0043] 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.

[0044] The compositions may be administered routinely, proactively, such as when at high risk, or when the need arises. The compositions may be administered with a meal, before a meal, after a meal, between meals, or at the onset of the disease or condition that the composition is intended to ameliorate.

[0045] (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.

[0046] The daily dose of the composition can be 0.1 g or more, preferably 0.3 g or more, more preferably 0.6 g or more, and even more preferably 1 g or more, of the active ingredient (the total amount if multiple active ingredients are included). The upper limit of the active ingredient per day, regardless of the lower limit, can be 30 g or less, 25 g or less, 20 g or less, 18 g or less, 17 mg or less, 16 g or less, or 15 g or less. When multiple active ingredients are included in the composition, the amount of active ingredient refers to the total amount of the active ingredients included.

[0047] Administration may be once a day or multiple times a day, for example, 2 to 10 divided doses. The amount of active ingredient per dose can be, for example, 0.01 g or more, preferably 0.3 g or more, more preferably 0.6 g or more, and even more preferably 1 g or more. The upper limit of the amount of active ingredient per dose, regardless of the lower limit, can be 20 g or less, 18 g or less, 16 g or less, 14 g or less, 12 g or less, 10 g or less, or 5 g or less.

[0048] 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 0.5% or more, preferably 1% or more, more preferably 2% or more, and even more preferably 3% or more. The upper limit of the active ingredient per solid content, regardless of the lower limit, can be 20% or less, 18% or less, 16% or less, 13% or less, or 10% or less. In the present invention, % means % by mass unless otherwise specified.

[0049] When the composition is a liquid such as a beverage, the content of the active ingredient can be, for example, 0.1% or more, preferably 0.3% or more, more preferably 0.6% or more, even more preferably 1% or more, and even more preferably 2% 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 15% or less, 10% or less, 8% or less, 6% or less, 4% or less, or 3% or less.

[0050] When the composition is in the form of a tablet or granular powder, the content of the active ingredient can be, for example, 10% or more, preferably 30% or more, more preferably 50% or more, and even more preferably 60% or more. When the composition is in the form of a tablet or granular powder, the upper limit of the content of the active ingredient can be 95% or less, 90% or less, or even 80% or less, regardless of the lower limit.

[0051] When the composition is a fermented milk such as yogurt, the content may be, for example, 2% or more, preferably 4% or more, and more preferably 10% or more. When the composition is a fermented milk such as yogurt, the upper limit of the content of the active ingredient may be 30% or less, 25% or less, or even 15% or less, regardless of the lower limit.

[0052] (Other Ingredients, Additives) The composition of the present invention 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), and sugars other than water-soluble dietary fiber. Sugars (glucose, 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.

[0053] In the present invention, the composition may contain a prebiotic other than water-soluble dietary fiber. 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 types of prebiotics other than water-soluble dietary fiber.

[0054] The prebiotics other than water-soluble dietary fiber are not particularly limited as long as they do not interfere with the effects of the active ingredients contained in the composition. Examples of prebiotics other than water-soluble dietary fiber include galactooligosaccharides, fructooligosaccharides, xylooligosaccharides, isomaltooligosaccharides, raffinose, lactulose, lactosucrose, soybean oligosaccharides, coffee oligosaccharides, insoluble dietary fiber, and gluconic acid.

[0055] The composition may further contain additives acceptable for food or pharmaceutical use, 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, dextrin, water-soluble dextran, water-soluble dextrin, corn starch, sodium carboxymethyl starch, pectin, xanthan gum, gum arabic, casein, gelatin, agar, glycerin, propylene glycol, polyethylene glycol, petrolatum, paraffin, stearyl alcohol, stearic acid, magnesium stearate, precipitated calcium carbonate, talc, human serum albumin, mannitol, sorbitol, lactose, sucralose, lactose hydrate, candy powder, white sugar, stevia, aspartame, acesulfame potassium, citric acid, lactic acid, malic acid, tartaric acid, phosphoric acid, acetic acid, fruit juice, and vegetable juice.

[0056] (Dosage Form / Form) The food composition of the present invention 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 may also be prepared in any form, such as a dairy product, supplement, confectionery, beverage, drink, seasoning, processed food, prepared dish, or soup. More specifically, the composition of the present invention may be in the form of a liquid diet, semi-liquid diet, jelly, gel, powder, infant formula, infant formula, liquid milk, powder or liquid milk for pregnant or nursing women, fermented milk, bar, mousse, chocolate, biscuit, ice cream, fermented milk, lactic acid bacteria drink, dairy drink, soft drink, tablet, cheese, bread, biscuit, cracker, pizza crust, food for the sick, nutritional food, frozen food, or processed food. It may also be in the form of a granule, powder, paste, or concentrated liquid for administration by mixing with beverages or foods. The granules and powder may be in the form of cubes or sticks (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.

[0057] 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.

[0058] The enteric coated formulation and pharmaceutical composition of the present invention can be made into any dosage form suitable for oral administration, such as solid formulations such as tablets, granules, powders, pills, and capsules; liquid formulations such as solutions, suspensions, and syrups; gels; and aerosols.

[0059] (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.

[0060] The composition of the present invention may be labeled with its intended use (application), and in one embodiment, it may be labeled with a recommendation that the composition be administered to a specific subject. Examples of subjects to be labeled are as described above. In one embodiment, the composition of the present invention may be labeled with the function of the composition or active ingredient or a usage method based on that function. For example, it may be labeled with the following: the ability to control the growth of Fusobacterium bacteria in the intestine, the ability to inhibit the growth of Fusobacterium bacteria in the intestine, the ability to treat diseases or conditions that are improved by controlling (preferably inhibiting) the growth of Fusobacterium bacteria in the intestine, the ability to use the composition as a prebiotic, the ability to use the composition as a synbiotic (a combination of probiotics and prebiotics), or the ability to treat inflammatory bowel disease and colorectal cancer. Note that a term such as "temporary" or "long-term" may be added to the beginning of each statement, as appropriate.

[0061] 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.

[0062] In one embodiment, the recommendation to consume the composition is displayed personally. Such a display can be provided via a document (whether written or electronic) addressed to the subject, the subject's tablet, smartphone, personal computer, or social networking site. Furthermore, such a display can be provided together with the results of any test or analysis, such as a gut flora test, a fecal substance test, or a fecal metabolome analysis, for the subject. This can also be utilized in precision nutrition (individualized nutrition, which refers to proposing an appropriate diet (nutrition) tailored to an individual's constitution).

[0063] In one aspect, 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 is provided, the method comprising the step of having the subject ingest a composition containing water-soluble dietary fiber based on the presence or amount of bacteria belonging to the genus Fusobacterium in the intestine of the subject. In another aspect, a method for suggesting to a subject to ingest a composition, a method for supporting a subject's diet, or a method for supporting a subject's health is provided, the method comprising the step of suggesting to a subject to ingest a composition containing water-soluble dietary fiber based on the presence or amount of bacteria belonging to the genus Fusobacterium in the intestine of the subject. These methods may include other steps, such as any of the following: collecting a fecal sample from the subject or having the subject collect a fecal sample; analyzing the presence or amount of Fusobacterium bacteria in the fecal sample; 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 product that is a composition containing water-soluble dietary fiber.

[0064] 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.

[0065] [Control of Fusobacterium spp. by water-soluble dietary fiber] (1) Preparation of test solution 90 mg of isomaltodextrin (Fibrixa (registered trademark), loss on drying 8.0% or less, pH 3.5 to 6.5 of 10% aqueous solution, dietary fiber (enzyme-HPLC method) 80.0% or more (per solids), nutritional values ​​(per 100 g): 216 kcal, carbohydrates 12.7 g, dietary fiber 82.7 g) was weighed out and dissolved in 1 mL of pure water at pH 4.5 in an anaerobic chamber. The solution was transferred through a 0.22 μm filter into a tube whose gas phase had been replaced in advance in an anaerobic chamber.

[0066] (2) Culture medium preparation: 12 mg of feces from six healthy adults was weighed into an Eppendorf tube in an anaerobic chamber, the gas phase of which had been replaced beforehand. Then, 500 μL of YCFA medium was added, homogenized, and YCFA medium was added to prepare fecal juice. The fecal juice had a fecal concentration of 0.1% (w / v). Then, 290 μL of fecal juice was added to each well of a 96-well plate, and 10 μL of the isomaltodextrin solution prepared in (1) (final concentration 0.3% (w / v)) or pure water was added to each well. These wells were designated as the test group (isomaltodextrin group) and the control group (water group), respectively.

[0067] (3) Culture The test and control groups prepared in (2) above were cultured for 16 hours at 37°C under anaerobic conditions. DNA was then extracted from each culture using a Maxwell® RSC Instrument (Promega).

[0068] (4) Measurement of bacterial count 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 in (3) 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 a standard.

[0069]

[0070] For stool samples in which Fusobacterium genus bacteria are not detected before culture, 1 × 10 Fusobacterium nucleatum JCM8532T was added at the start of culture. 4 cfu / ml was added.

[0071] As a result, the number of Fusobacterium bacteria was reduced in all cultures to which isomaltodextrin was added compared to the control (Figure 1).

[0072] These results suggest that isomaltodextrin may reduce the number of Fusobacterium bacteria.

[0073] Thus, water-soluble dietary fiber, such as isomaltodextrin, can be used to control Fusobacterium bacteria, particularly to reduce the number of bacteria in the intestinal flora. Fusobacterium bacteria have recently been suggested to be involved in colon cancer (e.g., Non-Patent Document 1), and F. nucleatum has been isolated from gastrointestinal infections such as appendicitis (Non-Patent Document 2), and has recently been suggested to be associated with inflammatory bowel disease (Non-Patent Documents 3, 4, and 5). Therefore, water-soluble dietary fiber, such as isomaltodextrin, can be used to treat these diseases.

[0074] [Production Example 1: Tablets] A tablet-type food composition for controlling bacteria belonging to the genus Fusobacterium was prepared using Fiberixa according to the following formulation in a standard manner. A similar food composition can be produced by adding indigestible dextrin or inulin instead of Fiberixa.

[0075] Granulated sugar 718 parts, concentrated fruit juice 50 parts, citric acid 60 parts, flavoring 20 parts, gelatin 10 parts, fiberixa 100 parts, calcium carbonate 15 parts, magnesium oxide 5 parts, potassium dihydrogen phosphate 22 parts

[0076] [Production Example 2: Powdered Drink] A powdered drink-type food composition for controlling bacteria belonging to the genus Fusobacterium was prepared using Fiberixa according to the following formulation in a standard manner. A similar food composition can be produced by adding indigestible dextrin or inulin instead of Fiberixa.

[0077] Anhydrous crystalline glucose 650 parts, citric acid granules 60 parts, tartaric acid 60 parts, baking soda granules 55 parts, flavor 25 parts, Fiberixa 100 parts, dolomite 50 parts

[0078] [Production Example 3: Beverage] Using Fiberixa, the ingredients were mixed and dissolved in the following proportions, sealed in a container, and heated at 90°C for 5 minutes to obtain a prebiotic beverage containing isomaltodextrin. Similar beverages can be produced by adding indigestible dextrin or inulin instead of Fiberixa.

[0079] Fiberixa 3 parts by weight, 5 times concentrated peach juice 2 parts by weight, sugar 5 parts by weight, citric acid 0.3 parts by weight, sodium citrate 0.05 parts by weight, water 89.65 parts by weight

[0080] [Production Example 4: Fermented Milk] A yogurt base mix is ​​prepared by mixing milk, a dairy product (derived from milk), and water so that the final product has a non-fat milk solids content of 9.5% and a milk fat content of 3.0%. The prepared yogurt base mix is ​​homogenized, sterilized by heating at 95°C for 5 minutes, and then cooled to approximately 40°C. This sterilized yogurt base mix is ​​inoculated with lactic acid bacteria Lactobacillus delbrueckii subsp. bulgaricus and lactic acid bacteria Streptococcus thermophilus, and the fermented product is added with isomaltodextrin to produce yogurt for controlling Fusobacterium bacteria in the intestinal flora, containing 200 mg of isomaltodextrin per product.

[0081] [Production Example 6: Capsules] Fiberixa is filled into enteric soft capsules at 200 mg each to produce a medicine for intestinal disorders, intestinal regulation (regulating bowel movements), or for improving loose stools, constipation, and abdominal distension.

[0082] Preparation Example 8: Enteric-coated supplement (tablets) A powder mixture of raw materials containing isomaltodextrin is tableted and coated to obtain tablets containing 100-400 mg of isomaltodextrin per tablet. The additives other than isomaltodextrin are corn starch, dextrin, lactose hydrate, precipitated calcium carbonate, candy powder, white sugar, talc, and magnesium stearate.

[0083] Preparation Example 7: Enteric-Release Supplement (Capsules) 100-400 mg of isomaltodextrin is filled 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.

[0084] 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.

[0085] [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, comprising water-soluble dietary fiber.

2. The composition according to claim 1, wherein the water-soluble dietary fiber is a low-viscosity water-soluble dietary fiber.

3. The composition according to claim 1, wherein the water-soluble dietary fiber is a hyperbranched α-glucan.

4. The composition according to claim 1, wherein the water-soluble dietary fiber is isomaltodextrin.

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

6. A composition according to any one of claims 1 to 5 for use as a prebiotic or synbiotic.

7. A composition according to any one of claims 1 to 5 for treating at least one of inflammatory bowel disease and colon cancer via control of bacteria belonging to the genus Fusobacterium.

8. A composition for treating colon cancer comprising α-glucan.

9. The composition according to claim 8, wherein the α-glucan is isomaltodextrin.

10. The composition according to claim 8 or 9, for administration to a subject having a bacterium belonging to the genus Fusobacterium in its intestines.

11. A pharmaceutical composition comprising isomaltodextrin.

12. A pharmaceutical composition according to claim 11 for use in treating intestinal diseases.

13. An enteric-coated preparation containing isomaltodextrin.

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