WO2020179871A1 - Eggerthella bacterial count suppressor - Google Patents

Eggerthella bacterial count suppressor Download PDF

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Publication number
WO2020179871A1
WO2020179871A1 PCT/JP2020/009451 JP2020009451W WO2020179871A1 WO 2020179871 A1 WO2020179871 A1 WO 2020179871A1 JP 2020009451 W JP2020009451 W JP 2020009451W WO 2020179871 A1 WO2020179871 A1 WO 2020179871A1
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Prior art keywords
kestose
eggerthella
bacteria
abscess
diabetes
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PCT/JP2020/009451
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French (fr)
Japanese (ja)
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吉弘 門田
勝昭 平野
巧 栃尾
明仁 遠藤
徹 工藤
悠一郎 西本
慎之介 村上
佳紀 水口
真嗣 福田
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株式会社メタジェン
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/702—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/02—Nasal agents, e.g. decongestants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents

Definitions

  • the present invention relates to the use of 1-kestose. Specifically, the present invention uses 1-kestose to control the number of Eggerthella bacteria, and 1-kestose to prevent or treat type 2 diabetes or various diseases involving Eggerthella bacteria. Regarding technology.
  • Eggerathella lentha a non-spore-forming anaerobic Gram-positive bacterium, belongs to the class Coriobacterium.
  • the bacterium is a constituent bacterium of the intestinal flora in healthy humans, on the other hand, in recent years, it has been implicated in infectious diseases caused by multiple microorganisms whose infectious agents are the digestive tract, and in severe disseminated diseases. It has been reported to be involved (Non-Patent Document 1; page 626, left column, paragraph 1-2).
  • Non-Patent Document 2 Specific examples of infectious diseases associated with the onset or exacerbation of Egacera lenta include bacteremia (Non-Patent Document 2), sinusitis (Non-Patent Document 3), purulent myositis (Non-Patent Document 4), and skin.
  • An abscess Non-Patent Document 5
  • a spondylodiscitis Non-Patent Document 6
  • a liver abscess Non-Patent Document 7 have been reported (Non-Patent Document 1; page 631, left column, second paragraph, Non-Patent Document 8). Page 475, left column, second paragraph).
  • the concentration of imidazole propionic acid which is a metabolite of histidine, is elevated in the plasma of patients with type 2 diabetes, imidazole propionic acid weakens glucose tolerance and insulin signaling, and Egasera renter uses imidazole propionic acid. It has been shown that the feces of diabetic patients who have the enzyme to be produced have a higher number of bacteria than those with normal glucose tolerance (Non-Patent Document 9: Page 949, left column, second paragraph). And FIGS. 2D, E, etc.). That is, it has been suggested that Eggerthella renter is involved in the onset or exacerbation of type 2 diabetes by producing imidazole propionic acid in vivo.
  • the present inventors considered that if the number of Eggerthella bacteria in the living body could be suppressed, it would be possible to contribute to the prevention and treatment of type 2 diabetes and various infectious diseases in which this bacterium is involved in the onset or exacerbation. That is, the present invention provides a technique capable of effectively suppressing the number of Eggerthella bacteria and thereby contributing to the prevention and treatment of type 2 diabetes and various infectious diseases in which the bacterium is involved in the onset or exacerbation. The purpose is to do.
  • a food composition containing 1-kestose as an active ingredient for controlling the number of bacteria of the genus Egacera is a food composition containing 1-kestose as an active ingredient for controlling the number of bacteria of the genus Egacera.
  • 1-Kestose used for the prevention or treatment of type 2 diabetes and diseases selected from bacteremia caused by Eggerthella bacteria, sinusitis, pyomyositis, cutaneous abscess, spinal discitis and liver abscess.
  • (5) Contains 1-kestose, which is used for the prevention or treatment of type 2 diabetes and diseases selected from bacteremia due to bacteria of the genus Egella, sinusitis, purulent myositis, skin abscess, spinal discitis and liver abscess Food to do.
  • a method for suppressing the number of bacteria of the genus Egacera which comprises administering 1-kestose to a human.
  • Diseases selected from type 2 diabetes and Eggerthella-induced bloodstream, sinusitis, pyomyositis, cutaneous abscess, spinal discitis and liver abscess, including administration of 1-kestose to humans. Methods for prevention or treatment.
  • Administering 1-kestose to a human suffering from type 2 diabetes and a disease selected from bacteremia due to bacteria of the genus Egella, sinusitis, purulent myositis, skin abscess, spinal discitis and liver abscess A method of treating the disease, including.
  • the number of Eggerthella bacteria in a living body can be effectively suppressed. Further, according to the present invention, by suppressing the number of Eggerthella bacteria, it is possible to contribute to the prevention or treatment of various diseases that develop or worsen due to the metabolic products of the bacteria or infection with the bacteria.
  • 1-kestose which is the active ingredient of the present invention, is a kind of oligosaccharide contained in vegetables and grains such as onion, garlic, barley, and rye, and is a substance having eating experience from ancient times.
  • Mutological toxicity test, acute toxicity test, subchronic toxicity test and chronic toxicity test the safety is extremely high (Food and Development, Vol. 49, No. 12, No. 12). 9 pages, 2014). Therefore, according to the present invention, it is possible to suppress the number of bacteria of the genus Egacera without causing any concern about safety.
  • Non-Patent Document 1 page 633, right column, second paragraph.
  • 1-kestose is a substance used as a food ingredient as described above, it is possible to suppress the number of Eggerthella bacteria without any concern about the appearance of resistant bacteria.
  • 1-kestose is highly water-soluble and has a good sweetness similar to sugar, so that it can be easily ingested as it is or as a seasoning such as a sweetener, and various foods. It can be easily blended in medicines, feeds, etc. Therefore, according to the present invention, it suppresses the number of Eggerthella bacteria that are highly safe and can be easily ingested on a daily basis as they are or by being easily mixed with various foods, pharmaceuticals, feeds and the like. Agents, food compositions, foods or pharmaceuticals for the purpose can be obtained.
  • Eggerthella bacterium refers to a microorganism belonging to the genus Eggerthella.
  • microorganisms include Eggerthella lentha, Eggerthella sinensis, Eggerthella brachy, Eggerthella hongkongensis, and Eggerthella infilm. , Eggerthella minutum, Eggerthella nodatum, Eggerthella saphenum, Eggerthella sulci, Eggerthella pneumonia , etc.
  • 1-Kestose is a trisaccharide oligosaccharide consisting of one molecule of glucose and two molecules of fructose.
  • 1-Kestose can be produced by performing an enzymatic reaction with an enzyme as disclosed in JP-A-58-201980 using sucrose as a substrate. Specifically, first, ⁇ -fructofuranosidase is added to a sucrose solution, and the mixture is allowed to stand at 37 ° C. to 50 ° C. for about 20 hours to carry out an enzymatic reaction to obtain a 1-kestose-containing reaction solution. By subjecting this 1-kestose-containing reaction solution to a chromatographic separation method as disclosed in Japanese Patent Application Laid-Open No. 2000-232878, 1-kestose and other sugars (glucose, fructose, sucrose, tetrasaccharide or more) can be used.
  • 1-kestose is contained in a commercially available fructooligosaccharide
  • 1-kestose may be used as it is, or 1-kestose may be separated and purified from the fructooligosaccharide by the above-mentioned method. That is, as the 1-kestose of the present invention, a 1-kestose-containing composition such as an oligosaccharide containing 1-kestose may be used.
  • the purity of 1-kestose is preferably 80% or more, more preferably 85% or more, and even more preferably 90% or more.
  • the "purity" of 1-kestose means the mass% of 1-kestose when the total mass of sugar is 100.
  • 1-Kestose can be used by administering it to humans or animals.
  • 1-Kestose exerts a function of suppressing the number of bacteria of the genus Egacera in the intestines of humans and animals.
  • the method of administering to humans and animals may be any method of reaching the intestine.
  • 1-kestose is added to an enteral nutritional supplement, and this is added by an enteral nutritional method via a tube inserted into the digestive tract such as the stomach or small intestine.
  • the method of administration can be exemplified.
  • 1-Kestose may be used in the form of a solid or aqueous solution consisting only of 1-Kestose, or in the form of an agent containing 1-Kestose, a drug, a food, a food composition, a food additive, a supplement or an animal feed. You may use it.
  • Examples of the dosage form of agents, pharmaceuticals, food additives, and supplements containing 1-kestose include powders, tablets, dragees, capsules, granules, dry syrups, liquids, syrups, drops, drinks, etc. Solid or liquid dosage forms can be mentioned.
  • the agents, pharmaceuticals, food additives, and supplements of the above dosage forms can be produced by methods known to those skilled in the art.
  • a powder 800 g of 1-kestose and 200 g of lactose are mixed well, and then 300 mL of 90% ethanol is added to moisten the powder. Subsequently, the wet powder is granulated, air-dried at 60 ° C. for 16 hours, and then granulated to obtain 1000 g (1-kestose content 800 mg / 1 g) of a powder having an appropriate fineness.
  • 1-Foods and food compositions containing kestose include confectionery and beverages, processed marine products and meat products, processed foods such as processed grain products, health foods, infant foods, dairy products, and edible granules. Examples include ordinary foods and drinks such as pastes, seasonings, and retort foods.
  • Foods and food compositions containing 1-kestose can be produced by adding 1-kestose to raw materials in a normal production process. The sweetness of 1-kestose is 30, and its taste, physical properties, and processability are close to those of sucrose. Therefore, in the manufacturing process, part or all of the sugar is replaced with 1-kestose, similar to sugar. It can be handled to produce various foods and food compositions.
  • the dose can be, for example, 0.04 g / kg body weight or more per day.
  • the dose is not limited to once a day, and may be divided into a plurality of doses.
  • "suppressing the number of bacteria" of an Eggerthella genus means suppressing an increase in the number of cells of the bacterium in any cell or tissue / organ of a living body.
  • the number of Eggerthella bacteria in the intestine is considered to correlate with the number of the bacteria in the feces or the contents of the cecum (hereinafter referred to as "feces, etc.”).
  • feces the number of the bacteria in the feces or the contents of the cecum.
  • a primer specific to Eggerthella bacteria is used as a sample of feces before and after administration of 1-kestose, or feces from an individual who has been administered and feces from an individual who has not been administered.
  • the 16S rRNA gene (16S rDNA) copy number is measured by performing the real-time PCR method used.
  • the 16S rDNA copy number can be used as an index of the bacterial count. Therefore, as a result of measuring the 16S rDNA copy number of the bacterium, if the 16S rDNA copy number in the sample after administration is smaller than that before administration, or the 16S rDNA copy number in the sample from the administered individual is not administered. If it is smaller than the above, it can be judged that 1-kestose has suppressed the number of bacteria of the genus Egacera.
  • Primers specific for 16S rDNA of Eggerthella bacteria can be designed based on known nucleotide sequences. For example, it can be designed based on the partial sequence of 16S rDNA of the genus Egacera (SEQ ID NO: 1) shown in Example 1 (3) described later.
  • the entire genome sequence of the reference strain of Eggerthella renter (VPI 0255, DSM 2243, ATCC 25559, JCM 9979) is available from GenBank accession number CP001726.1 (Saunders E et. Al., Stand Genomic Sci). 2009 Sep28;1(2):174-82.doi: 10.4056/sigs.33592.).
  • the 16SrDNA base sequence (SEQ ID NO: 2) is also disclosed in the whole genome base sequence information, and based on this, a primer specific to 16SrDNA of the genus Egacera can be designed.
  • Example 1 (3) described later stool or the like before and after administration of 1-kestose, or stool or the like from an individual who has been administered and stool or the like from an individual who has not been administered is used as a sample.
  • Bacterial-derived 16S rDNA in the sample is comprehensively analyzed by a generation sequencer, and the abundance ratio of sequence data having high homology with the sequence of the genus Eggerthella (SEQ ID NO: 1) is calculated. It can be said that the abundance ratio correlates with the 16S rDNA copy number of the Eggerthella bacterium, that is, the number of the bacterium.
  • Egacera lenta produces imidazole propionic acid from urocanic acid, and imidazole propione. Acids have been reported to impair glucose tolerance and insulin signaling (Non-Patent Document 9). Therefore, if the number of Escherichia bacteria in the intestine can be suppressed, it is possible to suppress the concentration of imidazole propionic acid in a person who may be affected by type 2 diabetes or a type 2 diabetes patient, which contributes to the prevention or treatment of type 2 diabetes. It is thought that it can be done. Therefore, type 1 diabetes can be prevented or treated by administering 1-kestose to humans or animals to suppress the number of bacteria of the genus Egacera. 1-Kestose can also be used in the manufacture of pharmaceuticals to prevent or treat type 2 diabetes.
  • 1-kestose can be used for the manufacture of a drug for suppressing the number of bacteria of the genus Egacera.
  • Bacteremia is defined as the state in which microorganisms are present in the bloodstream (Herbert M. Transitory bacteremia. Oral Surgery, Oral Medicine, Oral Pathology. 1954;7:609-615.). Bloodstream may be an asymptomatic temporary condition, but it may develop into a serious condition such as sepsis.
  • Non-patent document 2 Bacteremia due to Egacera lenta progresses to sepsis (Non-patent document 2; page 1278, first line, etc.), decubitus ulcer, bleeding and digestive tract obstruction, digestive tract perforation, diverticulum and pancreas, oviduct or It has been reported that symptoms such as abscesses in the ovary are present and that the mortality rate is as high as 20-36% when it develops (Non-Patent Document 8; page 477, right column 4. Discussion, paragraphs 1-2). ..
  • the number of Eggerthella bacteria in the intestine can be suppressed, the number of bacteria excreted in the environment and the number of bacteria invading the bloodstream from the intestine can be suppressed, which can contribute to the prevention or treatment of bloodstream disease caused by Eggerthella bacteria. it is conceivable that. Therefore, by administering 1-kestose to humans or animals to suppress the number of Eggerthella bacteria, it is possible to prevent or treat bloodstream diseases in which Eggerthella bacteria are involved in the onset or exacerbation. In addition, 1-kestose can be used in the manufacture of pharmaceuticals for preventing or treating bloodstream infections in which Eggerthella bacteria are involved in the onset or exacerbation.
  • Egasella lenta is associated with sinusitis (Non-Patent Document 3), purulent myositis (Non-Patent Document 4), skin abscess (Non-Patent Document 5), spinal discitis (Non-Patent Document 6) and It has been reported as a bacterium involved in the onset or exacerbation of liver abscess (Non-Patent Document 7) (Non-Patent Document 1; page 631, left column, second paragraph, non-patent document 8; page 475, left column, second paragraph). ..
  • 1-kestose is a pharmaceutical preparation for preventing or treating a disease selected from sinusitis, purulent myositis, cutaneous abscess, spondylodiscitis and liver abscess, which is involved in the onset or deterioration of bacteria of the genus Egacera. Can be used for.
  • Example 1 Examination of the inhibitory effect on the number of Eggerthella bacteria in vitro (1) Culture of fecal-containing microorganisms Previously reported ⁇ E Olano-Martin et al., Br J Nutr, Vol. 83, pp. 247-255, 2000 The anaerobic buffer and basal medium having the following compositions were prepared according to In addition, 1-kestose aqueous solution (final concentration 0.5% (w/v) was prepared. The anaerobic buffer, basic medium, 1-kestose aqueous solution, sterilized water and glass bottle were cleaned with nitrogen gas and then autoclave sterilized. went.
  • composition of anaerobic buffer / per 1 L >> (final pH: pH 7.0) Peptone 1 g, sodium chloride 8.5 g, cysteine hydrochloride 0.5 g.
  • Composition of basal medium / per 1 L >> (final pH: pH 7.0) Peptone 2 g, yeast extract 2 g, sodium chloride 0.1 g, dipotassium hydrogen phosphate 0.04 g, potassium dihydrogen phosphate 0.04 g, magnesium sulfate heptahydrate 0.01 g, calcium chloride dihydrate 0.
  • Feces were collected from 7 healthy persons (average age 25.6 years old, standard deviation ⁇ 6.4 years old, Body Mass Index range 22-25) and designated as Samples 1-7.
  • the healthy person had not taken antibiotics for more than 8 weeks before the feces were collected, and did not take prebiotics for more than 2 weeks before the feces had been collected.
  • the sample was immediately placed in an anaerobic jar (Aneropack, Mitsubishi Gas Chemical Company), and the following treatment was performed within 2 hours after collection.
  • DNA was extracted from this suspension using Magtration System 12GC (Precision System Science) and GC series MagDEA DNA 200 (Precision System Science). The concentration of DNA was measured using a spectrophotometer ND-1000 (NanDrop Technologies), and the concentration was adjusted to 10 ng/ ⁇ L, which was used as fecal-derived total DNA.
  • the base sequence of the PCR amplification product was decoded by the paired end method using the next-generation sequencer MiSeq (Illumina).
  • the obtained nucleotide sequence data those having low quality and those derived from the chimeric sequence were excluded.
  • a database was searched for the determined nucleotide sequence, and a taxon with an identity of 97% or higher was identified as one bacterial species (genus).
  • the sequence identified as the genus Eggerthella (partial sequence of 16S rDNA) is shown in SEQ ID NO: 1.
  • the abundance ratio was calculated as a percentage of the total number of leads in each group.
  • the quality check is in Reference 1, the chimera sequence check is ⁇ Robert C.
  • Table 1 shows the results of the genus Eggerthella in this comprehensive analysis.
  • the value of the abundance ratio is the average value in the samples 1 to 7.
  • Example 2 Examination of the inhibitory effect on the number of Eggerthella bacteria in vivo 38 healthy subjects (23-62 years old, male, weight 60.0-98.6 kg), 20 in the kestose group, 18 in the placebo group Divided into two groups of names.
  • the kestose group was orally ingested with 1-kestose and the placebo group with maltose daily.
  • the test period it was unknown to the subject whether he was ingesting 1-kestose or maltose.
  • the test period was 12 weeks, and the daily intake of 1-kestose or maltose was 10 g (about 0.17 to 0.10 g/kg body weight/day). Feces were collected at the beginning and end of the test period and cryopreserved at ⁇ 80 ° C.
  • Example 2 shows the results of the genus Eggerthella in this comprehensive analysis.
  • the value of the abundance ratio is the average value of 18 or 20 persons in each group.
  • the numerical range in each group is shown in parentheses.

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Abstract

[Problem] The purpose of the present invention is to make it possible to effectively suppress Eggerthella bacterial counts and thereby contribute to the prevention and treatment of type-2 diabetes and the various infections that are caused or worsened by Eggerthella. [Solution] An Eggerthella bacterial count suppressor that includes 1-kestose as an active ingredient. The present invention makes it possible to effectively suppress Eggerthella bacterial counts and can thereby contribute to the prevention or treatment of type-2 diabetes and infections, such as bacteremias, that are caused by Eggerthella. The Eggerthella bacterial count suppressor of the present invention is also highly safe, can be easily added to a variety of foods and pharmaceuticals, and is easy to take on a daily basis.　

Description

エガセラ属細菌の菌数抑制剤Eggerthella bacteria count suppressant
　本発明は、１－ケストースの用途に関する。詳細には、本発明は、１－ケストースを用いてエガセラ属細菌の菌数を抑制する技術、および、１－ケストースを用いて２型糖尿病もしくはエガセラ属細菌が関与する各種疾患を予防または治療する技術に関する。 The present invention relates to the use of 1-kestose. Specifically, the present invention uses 1-kestose to control the number of Eggerthella bacteria, and 1-kestose to prevent or treat type 2 diabetes or various diseases involving Eggerthella bacteria. Regarding technology.
　エガセラ属細菌（Eggerthella）のエガセラ・レンタ（Eggerthella lentha）は、非芽胞形成性の嫌気性グラム陽性細菌であり、コリオバクテリウム綱に属する。当該細菌は健康なヒトにおける腸内細菌叢の構成細菌であるが、その一方で、近年、消化管を感染源とする複数微生物による感染症に関与することや、重篤な播種性の疾患に関与することが報告されている（非特許文献１；第６２６頁左欄第１－２段落）。エガセラ・レンタが発症または悪化に関与する感染症として、具体的には、菌血症（非特許文献２）や副鼻腔炎（非特許文献３）、化膿性筋炎（非特許文献４）、皮膚膿瘍（非特許文献５）、脊椎椎間板炎（非特許文献６）、肝膿瘍（非特許文献７）が報告されている（非特許文献１；第６３１頁左欄第２段落、非特許文献８；第４７５頁左欄第２段落）。 Eggerathella lentha, a non-spore-forming anaerobic Gram-positive bacterium, belongs to the class Coriobacterium. Although the bacterium is a constituent bacterium of the intestinal flora in healthy humans, on the other hand, in recent years, it has been implicated in infectious diseases caused by multiple microorganisms whose infectious agents are the digestive tract, and in severe disseminated diseases. It has been reported to be involved (Non-Patent Document 1; page 626, left column, paragraph 1-2). Specific examples of infectious diseases associated with the onset or exacerbation of Egacera lenta include bacteremia (Non-Patent Document 2), sinusitis (Non-Patent Document 3), purulent myositis (Non-Patent Document 4), and skin. An abscess (Non-Patent Document 5), a spondylodiscitis (Non-Patent Document 6), and a liver abscess (Non-Patent Document 7) have been reported (Non-Patent Document 1; page 631, left column, second paragraph, Non-Patent Document 8). Page 475, left column, second paragraph).
　また、２型糖尿病患者の血漿中では、ヒスチジンの代謝産物であるイミダゾールプロピオン酸濃度が上昇していること、イミダゾールプロピオン酸は耐糖能およびインスリンシグナル伝達を弱めること、エガセラ・レンタはイミダゾールプロピオン酸を生成する酵素を持っており、かつ、糖尿病患者の糞便において、耐糖能が正常な者と比較して菌数が多いことが示されている（非特許文献９：第９４９頁左欄第２段落および図２Ｄ，Ｅなど）。すなわち、エガセラ・レンタは、生体内でイミダゾールプロピオン酸を生成することにより２型糖尿病の発症ないし悪化に関与することが示唆されている。 In addition, the concentration of imidazole propionic acid, which is a metabolite of histidine, is elevated in the plasma of patients with type 2 diabetes, imidazole propionic acid weakens glucose tolerance and insulin signaling, and Egasera renter uses imidazole propionic acid. It has been shown that the feces of diabetic patients who have the enzyme to be produced have a higher number of bacteria than those with normal glucose tolerance (Non-Patent Document 9: Page 949, left column, second paragraph). And FIGS. 2D, E, etc.). That is, it has been suggested that Eggerthella renter is involved in the onset or exacerbation of type 2 diabetes by producing imidazole propionic acid in vivo.
　そこで、本発明者らは、生体内のエガセラ属細菌の菌数を抑制できれば、２型糖尿病や、本細菌が発症または悪化に関与する各種感染症の予防や治療に貢献できると考えた。すなわち、本発明は、エガセラ属細菌の菌数を効果的に抑制することができ、もって２型糖尿病や本細菌が発症または悪化に関与する各種感染症の予防や治療に寄与しうる技術を提供することを目的とする。 Therefore, the present inventors considered that if the number of Eggerthella bacteria in the living body could be suppressed, it would be possible to contribute to the prevention and treatment of type 2 diabetes and various infectious diseases in which this bacterium is involved in the onset or exacerbation. That is, the present invention provides a technique capable of effectively suppressing the number of Eggerthella bacteria and thereby contributing to the prevention and treatment of type 2 diabetes and various infectious diseases in which the bacterium is involved in the onset or exacerbation. The purpose is to do.
　本発明者らは、鋭意研究の結果、１－ケストースが、腸内のエガセラ属細菌の菌数を顕著に抑制することを見出した。そこで、この知見に基づいて、下記の各発明を完成した。 As a result of diligent research, the present inventors have found that 1-kestose remarkably suppresses the number of Eggerthella bacteria in the intestine. Therefore, based on this finding, each of the following inventions was completed.
（１）１－ケストースを有効成分とする、エガセラ属細菌の菌数抑制剤。 (1) An agent for suppressing the number of Eggerthella bacteria containing 1-kestose as an active ingredient.
（２）２型糖尿病ならびにエガセラ属細菌による菌血症、副鼻腔炎、化膿性筋炎、皮膚膿瘍、脊椎椎間板炎および肝膿瘍から選択される疾患の予防または治療に用いられることを特徴とする、（１）に記載の菌数抑制剤。 (2) It is characterized by being used for the prevention or treatment of a disease selected from type 2 diabetes and bacteremia caused by Eggerthella bacteria, sinusitis, pyomyositis, skin abscess, spinal discitis and liver abscess. The bacterial count inhibitor according to (1).
（３）１－ケストースを有効成分とする、エガセラ属細菌の菌数抑制用食品組成物。 (3) A food composition containing 1-kestose as an active ingredient for controlling the number of bacteria of the genus Egacera.
（４）２型糖尿病ならびにエガセラ属細菌による菌血症、副鼻腔炎、化膿性筋炎、皮膚膿瘍、脊椎椎間板炎および肝膿瘍から選択される疾患の予防または治療に用いられる、１－ケストース。 (4) 1-Kestose used for the prevention or treatment of type 2 diabetes and diseases selected from bacteremia caused by Eggerthella bacteria, sinusitis, pyomyositis, cutaneous abscess, spinal discitis and liver abscess.
（５）２型糖尿病ならびにエガセラ属細菌による菌血症、副鼻腔炎、化膿性筋炎、皮膚膿瘍、脊椎椎間板炎および肝膿瘍から選択される疾患の予防または治療に用いられる、１－ケストースを含有する食品。 (5) Contains 1-kestose, which is used for the prevention or treatment of type 2 diabetes and diseases selected from bacteremia due to bacteria of the genus Egella, sinusitis, purulent myositis, skin abscess, spinal discitis and liver abscess Food to do.
（６）１－ケストースをヒトに投与することを含む、エガセラ属細菌の菌数を抑制するための方法。 (6) A method for suppressing the number of bacteria of the genus Egacera, which comprises administering 1-kestose to a human.
（７）１－ケストースをヒトに投与することを含む、２型糖尿病ならびにエガセラ属細菌による菌血症、副鼻腔炎、化膿性筋炎、皮膚膿瘍、脊椎椎間板炎および肝膿瘍から選択される疾患の予防または治療のための方法。 (7) Diseases selected from type 2 diabetes and Eggerthella-induced bloodstream, sinusitis, pyomyositis, cutaneous abscess, spinal discitis and liver abscess, including administration of 1-kestose to humans. Methods for prevention or treatment.
（８）２型糖尿病ならびにエガセラ属細菌による菌血症、副鼻腔炎、化膿性筋炎、皮膚膿瘍、脊椎椎間板炎および肝膿瘍から選択される疾患を患うヒトに、１－ケストースを投与することを含む、前記疾患を治療する方法。 (8) Administering 1-kestose to a human suffering from type 2 diabetes and a disease selected from bacteremia due to bacteria of the genus Egella, sinusitis, purulent myositis, skin abscess, spinal discitis and liver abscess A method of treating the disease, including.
（９）エガセラ属細菌の菌数を抑制するための医薬品の製造における１－ケストースの使用。 (9) Use of 1-kestose in the manufacture of pharmaceuticals to control the number of Eggerthella bacteria.
（１０）２型糖尿病ならびにエガセラ属細菌による菌血症、副鼻腔炎、化膿性筋炎、皮膚膿瘍、脊椎椎間板炎および肝膿瘍から選択される疾患の予防または治療のための医薬品の製造における１－ケストースの使用。 (10) 1- in the manufacture of pharmaceuticals for the prevention or treatment of types 2 diabetes and diseases selected from Eggerthella-induced bloodstream, sinusitis, pyomyositis, cutaneous abscess, spinal discitis and liver abscess. Use of Kestose.
　本発明によれば、生体におけるエガセラ属細菌の菌数を効果的に抑制することができる。また、本発明によれば、エガセラ属細菌の菌数を抑制することにより、当該細菌の代謝生成物あるいは当該細菌への感染によって発症あるいは悪化する各種疾患の予防や治療に寄与することができる。 According to the present invention, the number of Eggerthella bacteria in a living body can be effectively suppressed. Further, according to the present invention, by suppressing the number of Eggerthella bacteria, it is possible to contribute to the prevention or treatment of various diseases that develop or worsen due to the metabolic products of the bacteria or infection with the bacteria.
　また、本発明が有効成分とする１－ケストースは、タマネギやニンニク、大麦、ライ麦などの野菜や穀物にも含まれているオリゴ糖の一種であり、古来より食経験を有する物質であることや、変異原性試験、急性毒性試験、亜慢性毒性試験および慢性毒性試験のいずれにおいても毒性が認められていないことから、安全性は極めて高い（食品と開発、Ｖｏｌ．４９、Ｎｏ．１２、第９頁、２０１４年）。したがって、本発明によれば、安全性への懸念を全く生じさせずにエガセラ属細菌の菌数を抑制することができる。 In addition, 1-kestose, which is the active ingredient of the present invention, is a kind of oligosaccharide contained in vegetables and grains such as onion, garlic, barley, and rye, and is a substance having eating experience from ancient times. , Mutological toxicity test, acute toxicity test, subchronic toxicity test and chronic toxicity test, the safety is extremely high (Food and Development, Vol. 49, No. 12, No. 12). 9 pages, 2014). Therefore, according to the present invention, it is possible to suppress the number of bacteria of the genus Egacera without causing any concern about safety.
　特に、エガセラ属感染症の治療には、従来、各種の抗生物質が用いられているが、抗生物質の多用は耐性細菌の出現リスクを伴う（非特許文献１；第６３３頁右欄第２段落など）。この点、１－ケストースは上述のとおり食品成分に用いられる物質であるため、耐性細菌の出現を全く懸念することなく、エガセラ属細菌の菌数を抑制することができる。 In particular, various antibiotics have been conventionally used for the treatment of Eggerthella infectious diseases, but heavy use of antibiotics involves a risk of emergence of resistant bacteria (Non-Patent Document 1; page 633, right column, second paragraph). Such). In this respect, since 1-kestose is a substance used as a food ingredient as described above, it is possible to suppress the number of Eggerthella bacteria without any concern about the appearance of resistant bacteria.
　また、１－ケストースは、水溶性が高く、砂糖に似た良好な甘味質を有するため、そのまま、あるいは甘味料等の調味料として、日常的に簡便に摂取することができるほか、様々な食品や医薬品、飼料等に容易に配合することができる。したがって、本発明によれば、安全性が高く、そのまま、あるいは様々な食品や医薬品、飼料等に容易に配合して日常的に簡便に摂取することができる、エガセラ属細菌の菌数を抑制するための剤、食品組成物、食品または医薬品を得ることができる。 In addition, 1-kestose is highly water-soluble and has a good sweetness similar to sugar, so that it can be easily ingested as it is or as a seasoning such as a sweetener, and various foods. It can be easily blended in medicines, feeds, etc. Therefore, according to the present invention, it suppresses the number of Eggerthella bacteria that are highly safe and can be easily ingested on a daily basis as they are or by being easily mixed with various foods, pharmaceuticals, feeds and the like. Agents, food compositions, foods or pharmaceuticals for the purpose can be obtained.
　以下、本発明について詳細に説明する。 The present invention will be described in detail below.
　本発明において、「エガセラ属細菌」は、エガセラ属に属する微生物をいう。係る微生物としては、例えば、エガセラ・レンタ（Eggerthella lentha）やエガセラ・シネンシス（Eggerthella sinensis）、エガセラ・ブラキ(Eggerthella brachy)、エガセラ・ホンコンジェンシス(Eggerthella hongkongensis)、エガセラ・インフィルムム(Eggerthella infirmum)、エガセラ・ミヌタム(Eggerthella minutum)、エガセラ・ノダタム(Eggerthella nodatum)、エガセラ・サフェヌム(Eggerthella saphenum)、エガセラ・スルシ(Eggerthella sulci)、エガセラ・テヌエ(Eggerthella tenue)などを挙げることができる。 In the present invention, "Eggerthella bacterium" refers to a microorganism belonging to the genus Eggerthella. Examples of such microorganisms include Eggerthella lentha, Eggerthella sinensis, Eggerthella brachy, Eggerthella hongkongensis, and Eggerthella infilm. , Eggerthella minutum, Eggerthella nodatum, Eggerthella saphenum, Eggerthella sulci, Eggerthella tenue, etc.
　１－ケストースは、１分子のグルコースと２分子のフルクトースからなる三糖類のオリゴ糖である。 1-Kestose is a trisaccharide oligosaccharide consisting of one molecule of glucose and two molecules of fructose.
　１－ケストースは、スクロースを基質として、特開昭５８－２０１９８０号公報に開示されているような酵素による酵素反応を行うことにより作ることができる。具体的には、まず、β－フルクトフラノシダーゼをスクロース溶液に添加し、３７℃～５０℃で２０時間程度静置することにより酵素反応を行って、１－ケストース含有反応液を得る。この１－ケストース含有反応液を、特開２０００－２３２８７８号公報で開示されているようなクロマト分離法に供することよって、１－ケストースと他の糖（ブドウ糖、果糖、ショ糖、４糖以上のオリゴ糖）とを分離して精製し、高純度１－ケストース溶液を得る。続いて、この高純度１－ケストース溶液を濃縮した後、特公平６－７００７５号公報に開示されているような結晶化法で結晶化することにより、１－ケストースを結晶として得ることができる。 1-Kestose can be produced by performing an enzymatic reaction with an enzyme as disclosed in JP-A-58-201980 using sucrose as a substrate. Specifically, first, β-fructofuranosidase is added to a sucrose solution, and the mixture is allowed to stand at 37 ° C. to 50 ° C. for about 20 hours to carry out an enzymatic reaction to obtain a 1-kestose-containing reaction solution. By subjecting this 1-kestose-containing reaction solution to a chromatographic separation method as disclosed in Japanese Patent Application Laid-Open No. 2000-232878, 1-kestose and other sugars (glucose, fructose, sucrose, tetrasaccharide or more) can be used. (Oligosaccharide) is separated and purified to obtain a high-purity 1-kestose solution. Subsequently, this high-purity 1-kestose solution is concentrated and then crystallized by a crystallization method as disclosed in Japanese Patent Publication No. 6-70075, whereby 1-kestose can be obtained as crystals.
　また、１－ケストースは市販のフラクトオリゴ糖に含まれているため、これをそのまま、あるいは、フラクトオリゴ糖から上述の方法により１－ケストースを分離精製して用いてもよい。すなわち、本発明の１－ケストースとして、１－ケストースを含有するオリゴ糖などの１－ケストース含有組成物を用いてもよい。１－ケストース含有組成物を用いる場合、１－ケストースの純度は８０％以上であることが好ましく、８５％以上であることがより好ましく、９０％以上であることがさらに好ましい。なお、本発明において、１－ケストースの「純度」とは、糖の総質量を１００とした場合の、１－ケストースの質量％をいう。 Further, since 1-kestose is contained in a commercially available fructooligosaccharide, 1-kestose may be used as it is, or 1-kestose may be separated and purified from the fructooligosaccharide by the above-mentioned method. That is, as the 1-kestose of the present invention, a 1-kestose-containing composition such as an oligosaccharide containing 1-kestose may be used. When a 1-kestose-containing composition is used, the purity of 1-kestose is preferably 80% or more, more preferably 85% or more, and even more preferably 90% or more. In the present invention, the "purity" of 1-kestose means the mass% of 1-kestose when the total mass of sugar is 100.
　１－ケストースは、ヒトまたは動物に投与することにより使用することができる。１－ケストースは、ヒトや動物の腸内において、エガセラ属細菌の菌数を抑制する機能を発揮する。このことから、ヒトや動物に投与する方法としては、その腸内に到達させる方法であればよい。係る方法としては、例えば、経口摂取させる方法のほか、１－ケストースを経腸栄養剤に添加して、これを、胃や小腸などの消化管に挿入したチューブを経由して経腸栄養法により投与する方法を例示することができる。 1-Kestose can be used by administering it to humans or animals. 1-Kestose exerts a function of suppressing the number of bacteria of the genus Egacera in the intestines of humans and animals. From this, the method of administering to humans and animals may be any method of reaching the intestine. As such a method, for example, in addition to a method of ingesting orally, 1-kestose is added to an enteral nutritional supplement, and this is added by an enteral nutritional method via a tube inserted into the digestive tract such as the stomach or small intestine. The method of administration can be exemplified.
　１－ケストースは、１－ケストースのみからなる固体や水溶液の形態で用いてもよく、１－ケストースを含有する剤、医薬品、食品、食品組成物、食品添加剤、サプリメントまたは動物飼料などの形態で用いてもよい。 1-Kestose may be used in the form of a solid or aqueous solution consisting only of 1-Kestose, or in the form of an agent containing 1-Kestose, a drug, a food, a food composition, a food additive, a supplement or an animal feed. You may use it.
　１－ケストースを含有する剤や医薬品、食品添加剤、サプリメントの剤型としては、例えば、散剤、錠剤、糖衣剤、カプセル剤、顆粒剤、ドライシロップ剤、液剤、シロップ剤、ドロップ剤、ドリンク剤等の固形または液状の剤型を挙げることができる。 Examples of the dosage form of agents, pharmaceuticals, food additives, and supplements containing 1-kestose include powders, tablets, dragees, capsules, granules, dry syrups, liquids, syrups, drops, drinks, etc. Solid or liquid dosage forms can be mentioned.
　上記各剤型の剤や医薬品、食品添加剤、サプリメントは、当業者に公知の方法で製造することができる。例えば、散剤であれば、１－ケストース８００ｇおよび乳糖２００ｇをよく混合した後、９０％エタノール３００ｍＬを添加して湿潤させる。続いて、湿潤粉末を造粒した後、６０℃で１６時間通風乾燥し、その後、整粒して、適当な細かさの散剤１０００ｇ（１－ケストース含有量８００ｍｇ／１ｇ）を得ることができる。また、錠剤であれば、１－ケストース３００ｇ、粉末還元水飴３８０ｇ、コメデンプン１８０ｇおよびデキストリン１００ｇをよく混合した後、９０（ｖ／ｖ）％エタノール３００ｍＬを添加して湿潤させ、湿潤粉末を得る。この湿潤粉末を押し出し造粒した後、６０℃で１６時間通風乾燥して顆粒を得る。この顆粒を８５０μｍの篩を用いて整粒した後、顆粒４７０ｇに対してショ糖脂肪酸エステル５０ｇを添加して混合する。これを、ロータリー打錠機（６Ｂ－２、菊水製作所）に供して打錠することにより、直径８ｍｍ、重量２００ｍｇの錠剤５０００錠（１－ケストース含有量６０ｍｇ／１錠）を得ることができる。 The agents, pharmaceuticals, food additives, and supplements of the above dosage forms can be produced by methods known to those skilled in the art. For example, in the case of a powder, 800 g of 1-kestose and 200 g of lactose are mixed well, and then 300 mL of 90% ethanol is added to moisten the powder. Subsequently, the wet powder is granulated, air-dried at 60 ° C. for 16 hours, and then granulated to obtain 1000 g (1-kestose content 800 mg / 1 g) of a powder having an appropriate fineness. In the case of tablets, after thoroughly mixing 1-kestose (300 g), powdered reduced starch syrup (380 g), rice starch (180 g) and dextrin (100 g), 90 (v/v)% ethanol (300 mL) is added to wet the mixture to obtain a wet powder. After extruding and granulating this wet powder, it is air-dried at 60 ° C. for 16 hours to obtain granules. The granules are sized using a 850 μm sieve, and then 50 g of sucrose fatty acid ester is added to 470 g of the granules and mixed. By using this in a rotary tableting machine (6B-2, Kikusui Seisakusho) for tableting, 5000 tablets (1-kestose content 60 mg / tablet) having a diameter of 8 mm and a weight of 200 mg can be obtained.
　１－ケストースを含有する食品や食品組成物の形態としては、菓子や飲料、水産加工品や食肉加工品、穀物加工品などの加工食品、健康食品、乳幼児食品、乳製品、食用に供する顆粒、ペースト、調味料、レトルト食品などの通常の飲食物を挙げることができる。１－ケストースを含有する食品や食品組成物は、通常の製造過程で、原材料に１－ケストースを添加して製造することができる。１－ケストースの甘味度は３０で、その味質・物性・加工性はショ糖に近いことから、製造過程において、砂糖の一部または全部を１－ケストースに置き換えるなどして、砂糖と同様に扱って各種食品・食品組成物を製造することができる。 1-Foods and food compositions containing kestose include confectionery and beverages, processed marine products and meat products, processed foods such as processed grain products, health foods, infant foods, dairy products, and edible granules. Examples include ordinary foods and drinks such as pastes, seasonings, and retort foods. Foods and food compositions containing 1-kestose can be produced by adding 1-kestose to raw materials in a normal production process. The sweetness of 1-kestose is 30, and its taste, physical properties, and processability are close to those of sucrose. Therefore, in the manufacturing process, part or all of the sugar is replaced with 1-kestose, similar to sugar. It can be handled to produce various foods and food compositions.
　１－ケストースをヒトや動物に対して用いる場合の投与量（摂取量）としては、例えば、１日あたり０．０４ｇ／ｋｇ体重以上を挙げることができる。係る投与量は、１日１回に限らず、複数回に分割して投与してもよい。 When 1-kestose is used for humans and animals, the dose (intake) can be, for example, 0.04 g / kg body weight or more per day. The dose is not limited to once a day, and may be divided into a plurality of doses.
　本発明において、エガセラ属細菌の「菌数を抑制する」とは、生体のいずれかの細胞ないし組織・器官における当該細菌の細胞数の増加を抑制することをいう。 In the present invention, "suppressing the number of bacteria" of an Eggerthella genus means suppressing an increase in the number of cells of the bacterium in any cell or tissue / organ of a living body.
　例えば、腸におけるエガセラ属細菌の菌数は、糞便あるいは盲腸内容物（以下、「糞便等」という。）中の当該細菌の菌数と相関していると考えられるため、糞便等の中の当該細菌の菌数を計測することにより、腸においてエガセラ属細菌の菌数が抑制されたか否かを確認することができる。具体的には、例えば、１－ケストースの投与前後の糞便等、または、投与した個体からの糞便等と投与していない個体からの糞便等とを試料として、エガセラ属細菌に特異的なプライマーを用いたリアルタイムＰＣＲ法を行って１６Ｓ ｒＲＮＡ遺伝子（１６Ｓ ｒＤＮＡ）コピー数を計測する。当該細菌の１６Ｓ ｒＤＮＡコピー数と当該細菌の菌数とは相関関係にあるため、１６Ｓ ｒＤＮＡコピー数は、菌数の指標とすることができる。よって、当該細菌の１６Ｓ ｒＤＮＡコピー数を計測した結果、投与後の試料における１６Ｓ ｒＤＮＡコピー数が投与前よりも小さければ、あるいは、投与した個体からの試料における１６Ｓ ｒＤＮＡコピー数が投与していない個体よりも小さければ、１－ケストースによりエガセラ属細菌の菌数が抑制されたと判断することができる。 For example, the number of Eggerthella bacteria in the intestine is considered to correlate with the number of the bacteria in the feces or the contents of the cecum (hereinafter referred to as "feces, etc."). By measuring the number of bacteria, it is possible to confirm whether or not the number of Eggerthella bacteria is suppressed in the intestine. Specifically, for example, a primer specific to Eggerthella bacteria is used as a sample of feces before and after administration of 1-kestose, or feces from an individual who has been administered and feces from an individual who has not been administered. The 16S rRNA gene (16S rDNA) copy number is measured by performing the real-time PCR method used. Since there is a correlation between the 16S rDNA copy number of the bacterium and the bacterial number of the bacterium, the 16S rDNA copy number can be used as an index of the bacterial count. Therefore, as a result of measuring the 16S rDNA copy number of the bacterium, if the 16S rDNA copy number in the sample after administration is smaller than that before administration, or the 16S rDNA copy number in the sample from the administered individual is not administered. If it is smaller than the above, it can be judged that 1-kestose has suppressed the number of bacteria of the genus Egacera.
　エガセラ属細菌の１６Ｓ ｒＤＮＡに特異的なプライマーは、公知の塩基配列に基づいて設計することができる。例えば、後述する実施例１（３）に示すエガセラ属の１６Ｓ ｒＤＮＡの部分配列（配列番号１）に基づいて設計することができる。また、エガセラ・レンタの基準株（ＶＰＩ　０２５５、ＤＳＭ　２２４３、ＡＴＣＣ　２５５５９、ＪＣＭ　９９７９）の全ゲノム塩基配列はＧｅｎＢａｎｋアクセッション番号ＣＰ００１７２６．１から入手可能である（Saunders E et. al., Stand Genomic Sci. 2009 Sep 28;1(2):174-82. doi: 10.4056/sigs.33592.）。当該全ゲノム塩基配列情報においては、１６Ｓ ｒＤＮＡ塩基配列（配列番号２）も開示されており、これに基づいて、エガセラ属細菌の１６Ｓ ｒＤＮＡに特異的なプライマーを設計することができる。 Primers specific for 16S rDNA of Eggerthella bacteria can be designed based on known nucleotide sequences. For example, it can be designed based on the partial sequence of 16S rDNA of the genus Egacera (SEQ ID NO: 1) shown in Example 1 (3) described later. In addition, the entire genome sequence of the reference strain of Eggerthella renter (VPI 0255, DSM 2243, ATCC 25559, JCM 9979) is available from GenBank accession number CP001726.1 (Saunders E et. Al., Stand Genomic Sci). 2009 Sep28;1(2):174-82.doi: 10.4056/sigs.33592.). The 16SrDNA base sequence (SEQ ID NO: 2) is also disclosed in the whole genome base sequence information, and based on this, a primer specific to 16SrDNA of the genus Egacera can be designed.
　また、後述する実施例１（３）に示すように、１－ケストースの投与前後の糞便等、または、投与した個体からの糞便等と投与していない個体からの糞便等とを試料として、次世代シークエンサーにより試料中の細菌由来１６Ｓ ｒＤＮＡの網羅的解析を行い、エガセラ属の配列（配列番号１）と相同性の高い配列データの存在比率を算出する。当該存在比率は、エガセラ属細菌の１６Ｓ ｒＤＮＡコピー数、すなわち当該細菌の菌数と相関関係にあるといえる。よって、投与後の試料における当該配列データの存在比率が投与前よりも小さければ、あるいは、投与した個体の試料における当該配列データの存在比率が投与していない個体のものよりも小さければ、１－ケストースによりエガセラ属細菌の菌数が抑制されたと判断することができる。 Further, as shown in Example 1 (3) described later, stool or the like before and after administration of 1-kestose, or stool or the like from an individual who has been administered and stool or the like from an individual who has not been administered is used as a sample. Bacterial-derived 16S rDNA in the sample is comprehensively analyzed by a generation sequencer, and the abundance ratio of sequence data having high homology with the sequence of the genus Eggerthella (SEQ ID NO: 1) is calculated. It can be said that the abundance ratio correlates with the 16S rDNA copy number of the Eggerthella bacterium, that is, the number of the bacterium. Therefore, if the abundance ratio of the sequence data in the sample after administration is smaller than that before administration, or if the abundance ratio of the sequence data in the sample of the administered individual is smaller than that of the individual not administered, 1- It can be judged that the number of Eggerthella bacteria was suppressed by Kestose.
　上述のとおり、２型糖尿病患者の糞便すなわち腸内には、耐糖能正常者と比較してエガセラ・レンタが多く存在すること、エガセラ・レンタはウロカニン酸からイミダゾールプロピオン酸を生成すること、イミダゾールプロピオン酸は耐糖能およびインスリンシグナル伝達を弱めることが報告されている（非特許文献９）。よって、腸内のエガセラ属細菌の菌数を抑制できれば、２型糖尿病に罹患可能性のある者あるいは２型糖尿病患者におけるイミダゾールプロピオン酸の濃度を抑制できることから、２型糖尿病の予防または治療に寄与できると考えられる。したがって、１－ケストースをヒトや動物に投与して、エガセラ属細菌の菌数を抑制することにより、２型糖尿病を予防または治療することができる。また、１－ケストースは、２型糖尿病を予防または治療するための医薬品の製造に使用することができる。 As described above, in the feces of type 2 diabetic patients, that is, in the intestine, there is more Egacera lenta than in those with normal glucose tolerance, Egacera lenta produces imidazole propionic acid from urocanic acid, and imidazole propione. Acids have been reported to impair glucose tolerance and insulin signaling (Non-Patent Document 9). Therefore, if the number of Escherichia bacteria in the intestine can be suppressed, it is possible to suppress the concentration of imidazole propionic acid in a person who may be affected by type 2 diabetes or a type 2 diabetes patient, which contributes to the prevention or treatment of type 2 diabetes. It is thought that it can be done. Therefore, type 1 diabetes can be prevented or treated by administering 1-kestose to humans or animals to suppress the number of bacteria of the genus Egacera. 1-Kestose can also be used in the manufacture of pharmaceuticals to prevent or treat type 2 diabetes.
　また、１－ケストースは、エガセラ属細菌の菌数を抑制するための医薬品の製造に使用することができる。 Moreover, 1-kestose can be used for the manufacture of a drug for suppressing the number of bacteria of the genus Egacera.
　菌血症は、微生物が血流中に存在する状態として定義される（Herbert M. Transitory bacteremia. Oral Surgery, Oral Medicine, Oral Pathology. 1954;7:609-615.）。菌血症は無症状な一時的な状態である場合もあるが、敗血症などの重篤な状態に進展する場合がある。エガセラ・レンタによる菌血症は、敗血症に進行し（非特許文献２；第１２７８頁第１行目など）、褥瘡性潰瘍、出血や消化管閉塞、消化管穿孔、憩室や膵臓、卵管ないし卵巣における膿瘍といった症状を呈することや、発症した場合には死亡率が２０－３６％と高いことが報告されている（非特許文献８；第４７７頁右欄４．Ｄｉｓｃｕｓｓｉｏｎ第１－２段落）。腸内のエガセラ属細菌の菌数を抑制できれば、環境中に排泄される菌数や腸から血流に侵入する菌数を抑制できることから、エガセラ属細菌による菌血症の予防または治療に寄与できると考えられる。したがって、１－ケストースをヒトや動物に投与して、エガセラ属細菌の菌数を抑制することにより、エガセラ属細菌が発症または悪化に関与する菌血症を予防または治療することができる。また、１－ケストースは、エガセラ属細菌が発症もしくは悪化に関与する菌血症を、予防または治療するための医薬品の製造に使用することができる。 Bacteremia is defined as the state in which microorganisms are present in the bloodstream (Herbert M. Transitory bacteremia. Oral Surgery, Oral Medicine, Oral Pathology. 1954;7:609-615.). Bloodstream may be an asymptomatic temporary condition, but it may develop into a serious condition such as sepsis. Bacteremia due to Egacera lenta progresses to sepsis (Non-patent document 2; page 1278, first line, etc.), decubitus ulcer, bleeding and digestive tract obstruction, digestive tract perforation, diverticulum and pancreas, oviduct or It has been reported that symptoms such as abscesses in the ovary are present and that the mortality rate is as high as 20-36% when it develops (Non-Patent Document 8; page 477, right column 4. Discussion, paragraphs 1-2). .. If the number of Eggerthella bacteria in the intestine can be suppressed, the number of bacteria excreted in the environment and the number of bacteria invading the bloodstream from the intestine can be suppressed, which can contribute to the prevention or treatment of bloodstream disease caused by Eggerthella bacteria. it is conceivable that. Therefore, by administering 1-kestose to humans or animals to suppress the number of Eggerthella bacteria, it is possible to prevent or treat bloodstream diseases in which Eggerthella bacteria are involved in the onset or exacerbation. In addition, 1-kestose can be used in the manufacture of pharmaceuticals for preventing or treating bloodstream infections in which Eggerthella bacteria are involved in the onset or exacerbation.
　また、上述のとおり、エガセラ・レンタは、副鼻腔炎（非特許文献３）、化膿性筋炎（非特許文献４）、皮膚膿瘍（非特許文献５）、脊椎椎間板炎（非特許文献６）および肝膿瘍（非特許文献７）の発症または悪化に関与する細菌として報告されている（非特許文献１；第６３１頁左欄第２段落、非特許文献８；第４７５頁左欄第２段落）。腸内のエガセラ属細菌の菌数を抑制できれば、これらの感染症の予防または治療に寄与できると考えられる。したがって、１－ケストースをヒトや動物に投与して、エガセラ属細菌の菌数を抑制することにより、エガセラ属細菌が発症または悪化に関与する、副鼻腔炎、化膿性筋炎、皮膚膿瘍、脊椎椎間板炎および肝膿瘍から選択される疾患を、予防または治療することができる。また、１－ケストースは、エガセラ属細菌が発症または悪化に関与する、副鼻腔炎、化膿性筋炎、皮膚膿瘍、脊椎椎間板炎および肝膿瘍から選択される疾患を予防または治療するための医薬品の製造に使用することができる。 In addition, as described above, Egasella lenta is associated with sinusitis (Non-Patent Document 3), purulent myositis (Non-Patent Document 4), skin abscess (Non-Patent Document 5), spinal discitis (Non-Patent Document 6) and It has been reported as a bacterium involved in the onset or exacerbation of liver abscess (Non-Patent Document 7) (Non-Patent Document 1; page 631, left column, second paragraph, non-patent document 8; page 475, left column, second paragraph). .. It is considered that if the number of bacteria of the genus Egacera in the intestine can be suppressed, it can contribute to the prevention or treatment of these infections. Therefore, sinusitis, purulent myositis, skin abscess, spinal disc involving the involvement of Egella bacteria in the onset or exacerbation of Egacera bacteria by administering 1-kestose to humans or animals to suppress the number of Egacella bacteria A disease selected from inflammation and liver abscess can be prevented or treated. In addition, 1-kestose is a pharmaceutical preparation for preventing or treating a disease selected from sinusitis, purulent myositis, cutaneous abscess, spondylodiscitis and liver abscess, which is involved in the onset or deterioration of bacteria of the genus Egacera. Can be used for.
　以下、本発明について、各実施例に基づいて説明する。なお、本発明の技術的範囲は、これらの実施例によって示される特徴に限定されない。また、本実施例においては、「１－ケストース」として、純度９８質量％以上で１－ケストースを含有する組成物（物産フードサイエンス社）を用いた。 Hereinafter, the present invention will be described based on each embodiment. The technical scope of the present invention is not limited to the features shown in these examples. Further, in this example, as "1-kestose", a composition containing 1-kestose with a purity of 98% by mass or more (Busan Food Science Co., Ltd.) was used.
＜実施例１＞生体外でのエガセラ属細菌の菌数抑制作用の検討
（１）糞便含有微生物の培養
　既報〈Ｅ　Ｏｌａｎｏ－Ｍａｒｔｉｎら、Ｂｒ　Ｊ　Ｎｕｔｒ、第８３巻、第２４７－２５５頁、２０００年〉に従って下記の組成の嫌気緩衝液および基本培地を調製した。また、１－ケストース水溶液（終濃度０．５％（ｗ／ｖ）を調製した。嫌気緩衝液、基本培地、１－ケストース水溶液、滅菌水および硝子瓶は窒素ガスで浄化した後、オートクレーブ滅菌を行った。
　《嫌気緩衝液の組成／１Ｌあたり》（最終ｐＨ：ｐＨ７．０）
　ペプトン１ｇ、塩化ナトリウム８．５ｇ、システイン塩酸塩０．５ｇ。
　《基本培地の組成／１Ｌあたり》（最終ｐＨ：ｐＨ７．０）
　ペプトン２ｇ、酵母抽出物２ｇ、塩化ナトリウム０．１ｇ、リン酸水素二カリウム０．０４ｇ、リン酸二水素カリウム０．０４ｇ、硫酸マグネシウム七水和物０．０１ｇ、塩化カルシウム二水和物０．０１ｇ、炭酸水素ナトリウム２ｇ、システイン塩酸塩０．５ｇ、胆汁酸塩０．５ｇ、ヘミン０．００５ｇ、Ｔｗｅｅｎ　８０　２ｍＬ、フィロキノン１０μＬ、レザズリンナトリウム。 <Example 1> Examination of the inhibitory effect on the number of Eggerthella bacteria in vitro (1) Culture of fecal-containing microorganisms Previously reported <E Olano-Martin et al., Br J Nutr, Vol. 83, pp. 247-255, 2000 The anaerobic buffer and basal medium having the following compositions were prepared according to In addition, 1-kestose aqueous solution (final concentration 0.5% (w/v) was prepared. The anaerobic buffer, basic medium, 1-kestose aqueous solution, sterilized water and glass bottle were cleaned with nitrogen gas and then autoclave sterilized. went.
<< Composition of anaerobic buffer / per 1 L >> (final pH: pH 7.0)
Peptone 1 g, sodium chloride 8.5 g, cysteine hydrochloride 0.5 g.
<< Composition of basal medium / per 1 L >> (final pH: pH 7.0)
Peptone 2 g, yeast extract 2 g, sodium chloride 0.1 g, dipotassium hydrogen phosphate 0.04 g, potassium dihydrogen phosphate 0.04 g, magnesium sulfate heptahydrate 0.01 g, calcium chloride dihydrate 0. 01 g, sodium hydrogen carbonate 2 g, cysteine hydrochloride 0.5 g, bile salt 0.5 g, hemin 0.005 g, Tween 80 2 mL, phylloquinone 10 μL, resazurin sodium.
　７名の健常者（平均年齢２５．６歳、標準偏差±６．４歳、体格指数（Body Mass Index）範囲２２～２５）から糞便を採取し、検体１～７とした。なお、当該健常者は、糞便採取時から以前８週間以上抗生剤を摂取しておらず、糞便採取時から以前２週間以上プレバイオティクスを摂取していない。検体は速やかに嫌気ジャー（アネロパック、三菱ガス化学）に入れ、採取から２時間以内に下記の処理を行った。 Feces were collected from 7 healthy persons (average age 25.6 years old, standard deviation ±6.4 years old, Body Mass Index range 22-25) and designated as Samples 1-7. The healthy person had not taken antibiotics for more than 8 weeks before the feces were collected, and did not take prebiotics for more than 2 weeks before the feces had been collected. The sample was immediately placed in an anaerobic jar (Aneropack, Mitsubishi Gas Chemical Company), and the following treatment was performed within 2 hours after collection.
　各検体から適量を取って培養前検体とし、－８０℃にて凍結保存した。また、残りの各検体を９ｍＬの嫌気緩衝液に入れて１０倍（ｗ／ｖ）に希釈した後、窒素ガスを満たした環境下で２倍濃度の基本培地４０ｍＬを入れた硝子瓶に移した。このうち５ｍＬを、同様の嫌気環境下の硝子瓶にて、等量の１－ケストース水溶液または滅菌水と混合した。これを３７℃で４８時間静置培養した後、１２０００×ｇで５分間遠心分離を行い、沈殿物を回収して、これを培養後検体とし、－８０℃にて凍結保存した。 An appropriate amount was taken from each sample to prepare a pre-culture sample, which was cryopreserved at -80 ° C. In addition, each of the remaining samples was placed in 9 mL of anaerobic buffer, diluted 10-fold (w / v), and then transferred to a glass bottle containing 40 mL of 2-fold concentrated basal medium in an environment filled with nitrogen gas. .. Of these, 5 mL was mixed with an equal volume of 1-kestose aqueous solution or sterile water in a glass bottle under the same anaerobic environment. This was statically cultured at 37 ° C. for 48 hours, then centrifuged at 12000 × g for 5 minutes, and the precipitate was collected, which was used as a sample after culturing and cryopreserved at −80 ° C.
（２）総ＤＮＡの抽出
　〈Ｓｈｕｎｓｕｋｅ Ｔａｋａｈａｓｈｉら、ＰＬｏｓＯＮＥ、第９巻、第８号、ｅ１０５５９２、２０１４年８月：参考文献１〉に記載の方法に従って、本実施例１（１）の培養前検体および培養後検体から総ＤＮＡを抽出した。具体的には、まず、４Ｍのグアニジンチオシアネート、１００ｍＭのトリスＨＣｌ（ｐＨ９．０）および４０ｍＭのＥＤＴＡを含む水溶液に、氷上で融解した培養前検体または培養後検体１００ｍｇを懸濁し、FastPrep FP100A(MP Biomedicals)を用いてジルコニアビーズで粉砕して懸濁液を得た。Magtration System 12GC（Precision System Science）およびGC series MagDEA DNA 200（Precision System Science）を用いて、この懸濁液からＤＮＡを抽出した。ＤＮＡの濃度を分光測光器ND-1000（NanDrop Technologies）を用いて測定し、１０ｎｇ／μＬとなるように調製して、これを糞便由来総ＤＮＡとした。 (2) Extraction of total DNA According to the method described in <Shunsuke Takahashi et al., PLOS ONE, Vol. 9, No. 8, e105592, August 2014: Reference 1>, the pre-culture sample of Example 1 (1). And total DNA was extracted from the sample after culturing. Specifically, first, 100 mg of a pre-culture sample or a post-culture sample melted on ice is suspended in an aqueous solution containing 4 M guanidine thiocyanate, 100 mM Tris HCl (pH 9.0) and 40 mM EDTA, and FastPrep FP100A (MP). Biomedicals) was used to grind with zirconia beads to obtain a suspension. DNA was extracted from this suspension using Magtration System 12GC (Precision System Science) and GC series MagDEA DNA 200 (Precision System Science). The concentration of DNA was measured using a spectrophotometer ND-1000 (NanDrop Technologies), and the concentration was adjusted to 10 ng/μL, which was used as fecal-derived total DNA.
（３）糞便中の細菌由来ＤＮＡの網羅的解析
　上記参考文献１に記載の方法に従い、本実施例１（１）の培養前後の糞便に含まれる細菌の種類と存在比率の網羅的解析を行った。すなわち、まず、本実施例１（２）の糞便来総ＤＮＡを鋳型として、下記配列番号３および４のユニバーサルプライマーを用いてＤｕａｌ－ｉｎｄｅｘ法（Ｈｉｓａｄａ Ｔａｋａｙｏｓｈｉら、Ａｒｃｈ Ｍｉｃｒｏｂｉｏｌ、１９７巻、第７号、第９１９‐３４頁、２０１５年６月）によりポリメラーゼ連鎖反応（ＰＣＲ）を行い、細菌由来の１６Ｓ ｒＤＮＡのＶ３－Ｖ４領域を増幅した。
　フォワードプライマー（３４１ｆ）：ＣＣＴＡＣＧＧＧＡＧＧＣＡＧＣＡＧ（配列番号３）
　リバースプライマー（Ｒ８０６）：ＧＧＡＣＴＡＣＨＶＧＧＧＴＷＴＣＴＡＡＴ（配列番号４） (3) Comprehensive analysis of bacterial-derived DNA in feces According to the method described in Reference 1 above, a comprehensive analysis of the types and abundance ratios of bacteria contained in feces before and after culturing in Example 1 (1) was performed. It was. That is, first, the dual-index method (Hisada Takayoshi et al., Arch Microbiol, Vol. 197, No. 7) was performed using the fecal total DNA of Example 1 (2) as a template and the universal primers of SEQ ID NOs: 3 and 4 below. , 919-34, June 2015) to amplify the V3-V4 region of 16S rDNA derived from bacteria by polymerase chain reaction (PCR).
Forward primer (341f): CCTACGGGAGGCAGCAG (SEQ ID NO: 3)
Reverse primer (R806): GGACTACHVGGGWTTCTAAT (SEQ ID NO: 4)
　続いて、次世代シークエンサーＭｉＳｅｑ（Illumina）によりＰＣＲ増幅産物の塩基配列をペアエンド法により解読した。得られた塩基配列データはデータの質（クオリティ）が低いものおよびキメラ配列由来のものを排除した。決定した塩基配列についてデータベースによる検索を行い、同一性が９７％以上で検出される分類群を１菌種（属）として同定した。エガセラ属として同定された配列（１６Ｓ ｒＤＮＡの部分配列）を配列番号１に示す。存在比率は、総リード数に占める各群のリード数の割合を百分率として算出した。なお、クオリティのチェックは参考文献１に、キメラ配列のチェックは〈Robert C. Edgarら、BIOINFORMATICS、第２７巻、１６号、第２１９４－２２００頁、２０１１年６月〉に、データベースによる検索は〈Ｈｉｓａｄａ Ｔａｋａｙｏｓｈｉら、Ａｒｃｈ Ｍｉｃｒｏｂｉｏｌ、１９７巻、第７号、第９１９‐３４頁、２０１５年６月〉に、それぞれ記載の方法に準じて行った。この網羅的解析におけるエガセラ属の結果を表１に示す。なお、表１において、存在比率の値は、検体１～７における平均値である。
Figure JPOXMLDOC01-appb-T000001
 Subsequently, the base sequence of the PCR amplification product was decoded by the paired end method using the next-generation sequencer MiSeq (Illumina). As for the obtained nucleotide sequence data, those having low quality and those derived from the chimeric sequence were excluded. A database was searched for the determined nucleotide sequence, and a taxon with an identity of 97% or higher was identified as one bacterial species (genus). The sequence identified as the genus Eggerthella (partial sequence of 16S rDNA) is shown in SEQ ID NO: 1. The abundance ratio was calculated as a percentage of the total number of leads in each group. The quality check is in Reference 1, the chimera sequence check is <Robert C. Edgar et al., BIOINFORMATICS, Vol. 27, No. 16, No. 2194-2200, June 2011>, and the database search is < Hizada Takayoshi et al., Arch Microbiol, Vol. 197, No. 7, pp. 919-34, June 2015>, respectively, according to the method described. Table 1 shows the results of the genus Eggerthella in this comprehensive analysis. In Table 1, the value of the abundance ratio is the average value in the samples 1 to 7.
Figure JPOXMLDOC01-appb-T000001
　表１に示すように、１－ケストース存在下で培養した場合、エガセラ属細菌の１６Ｓ ｒＤＮＡの存在比率は、培養前検体では０．３０％であったのに対して、培養後検体では０．０７％であり、培養前と比較して１／４．２倍と顕著に減少した。これに対して、１－ケストース非存在下で培養した場合、培養後検体では１．６０％であり、培養前と比較して５．３倍と顕著に増加した。この結果から、１－ケストースの存在下では、糞便中のエガセラ属細菌の１６Ｓ ｒＤＮＡの存在比率が減少することが明らかになった。すなわち、１－ケストースは、生体外においてエガセラ属細菌の菌数を抑制する効果を有することが明らかになった。 As shown in Table 1, when cultured in the presence of 1-kestose, the abundance ratio of 16S rDNA of Eggerthella spp. Was 0.30% in the pre-culture sample, whereas it was 0. It was 07%, which was a significant decrease of 1 / 4.2 times as compared with that before culturing. On the other hand, when culturing in the absence of 1-kestose, it was 1.60% in the sample after culturing, which was a remarkable increase of 5.3 times as compared with that before culturing. From this result, it was clarified that the abundance ratio of 16S rDNA of Eggerthella bacteria in feces decreased in the presence of 1-kestose. That is, it was revealed that 1-kestose has an effect of suppressing the number of bacteria of the genus Egacera in vitro.
＜実施例２＞生体内でのエガセラ属細菌の菌数抑制作用の検討
　健常者（２３～６２歳、男性、体重６０．０～９８．６ｋｇ）３８名を、ケストース群２０名、プラセボ群１８名の２つの群に分けた。ケストース群には１－ケストースを、プラセボ群にはマルトースを、試験期間中、毎日経口摂取させた。なお、試験期間中、被検者には、１－ケストースまたはマルトースのいずれを摂取しているのか不明な状態とした。試験期間は１２週間、１－ケストースまたはマルトースの１日の摂取量は１０ｇ（約０．１７～０．１０ｇ／ｋｇ体重／日）とした。試験期間の開始時および終了時に糞便を採取し、－８０℃にて凍結保存した。 <Example 2> Examination of the inhibitory effect on the number of Eggerthella bacteria in vivo 38 healthy subjects (23-62 years old, male, weight 60.0-98.6 kg), 20 in the kestose group, 18 in the placebo group Divided into two groups of names. The kestose group was orally ingested with 1-kestose and the placebo group with maltose daily. During the test period, it was unknown to the subject whether he was ingesting 1-kestose or maltose. The test period was 12 weeks, and the daily intake of 1-kestose or maltose was 10 g (about 0.17 to 0.10 g/kg body weight/day). Feces were collected at the beginning and end of the test period and cryopreserved at −80 ° C.
　採取した糞便から、実施例１（２）に記載の方法により総ＤＮＡを抽出した。続いて、実施例１（３）に記載の方法により糞便中の細菌由来ＤＮＡの網羅的解析を行った。この網羅的解析におけるエガセラ属の結果を表２に示す。なお、表２において、存在比率の値は、各群１８名または２０名における平均値である。また、括弧内に、各群における数値範囲を示す。
Figure JPOXMLDOC01-appb-T000002
 Total DNA was extracted from the collected feces by the method described in Example 1 (2). Subsequently, a comprehensive analysis of bacterial DNA in feces was performed by the method described in Example 1 (3). Table 2 shows the results of the genus Eggerthella in this comprehensive analysis. In Table 2, the value of the abundance ratio is the average value of 18 or 20 persons in each group. In addition, the numerical range in each group is shown in parentheses.
Figure JPOXMLDOC01-appb-T000002
　表２に示すように、プラセボ群では、エガセラ属細菌の１６Ｓ ｒＤＮＡの存在比率は試験開始時に０．１００％であったのに対して試験終了時には０．１７９％であり、増加していた。これに対して、ケストース群では、エガセラ属細菌の１６Ｓ ｒＤＮＡの存在比率は試験開始時に０．０８９％であったのに対して試験終了時には０．０６７％であり、減少していた。この結果から、１－ケストースの摂取により、糞便中のエガセラ属細菌の１６Ｓ ｒＤＮＡの存在比率が減少することが明らかになった。すなわち、１－ケストースは、生体内においてエガセラ属細菌の菌数を抑制する効果を有することが明らかになった。 As shown in Table 2, in the placebo group, the abundance ratio of 16S rDNA of Eggerthella spp. Was 0.100% at the start of the test and 0.179% at the end of the test, which was an increase. On the other hand, in the Kestose group, the abundance ratio of 16S rDNA of Eggerthella bacteria was 0.089% at the start of the test, whereas it was 0.067% at the end of the test, which was a decrease. From this result, it was clarified that ingestion of 1-kestose reduces the abundance ratio of 16S rDNA of Eggerthella bacteria in feces. That is, it was revealed that 1-kestose has an effect of suppressing the number of Egella bacteria in vivo.

Claims (10)

  1. 　１－ケストースを有効成分とする、エガセラ属細菌の菌数抑制剤。 1-Kestose is an active ingredient, an agent for suppressing the number of Eggerthella bacteria.
  2. 　２型糖尿病ならびにエガセラ属細菌による菌血症、副鼻腔炎、化膿性筋炎、皮膚膿瘍、脊椎椎間板炎および肝膿瘍から選択される疾患の予防または治療に用いられることを特徴とする、請求項１に記載の菌数抑制剤。 Claim 1 characterized by being used for the prevention or treatment of type 2 diabetes and diseases selected from Eggerthella-induced sepsis, sinusitis, pyomyositis, cutaneous abscess, spinal discitis and liver abscess. Bacterial count inhibitor according to.
  3. 　１－ケストースを有効成分とする、エガセラ属細菌の菌数抑制用食品組成物。 A food composition for controlling the number of bacteria of the genus Egacera, which contains 1-kestose as an active ingredient.
  4. 　２型糖尿病ならびにエガセラ属細菌による菌血症、副鼻腔炎、化膿性筋炎、皮膚膿瘍、脊椎椎間板炎および肝膿瘍から選択される疾患の予防または治療に用いられる、１－ケストース。 1-Kestose used for the prevention or treatment of type 2 diabetes and diseases selected from bacteremia caused by Eggerthella bacteria, sinusitis, pyomyositis, cutaneous abscess, spinal discitis and liver abscess.
  5. 　２型糖尿病ならびにエガセラ属細菌による菌血症、副鼻腔炎、化膿性筋炎、皮膚膿瘍、脊椎椎間板炎および肝膿瘍から選択される疾患の予防または治療に用いられる、１－ケストースを含有する食品。 A food containing 1-kestose, which is used for the prevention or treatment of type 2 diabetes and a disease selected from bacteremia due to bacteria of the genus Egella, sinusitis, purulent myositis, skin abscess, spinal discitis and liver abscess.
  6. 　１－ケストースをヒトに投与することを含む、エガセラ属細菌の菌数を抑制するための方法。 A method for suppressing the number of bacteria of the genus Egacera, which comprises administering 1-kestose to a human.
  7. 　１－ケストースをヒトに投与することを含む、２型糖尿病ならびにエガセラ属細菌による菌血症、副鼻腔炎、化膿性筋炎、皮膚膿瘍、脊椎椎間板炎および肝膿瘍から選択される疾患の予防または治療のための方法。 Prevention or treatment of type 2 diabetes mellitus including administration of 1-kestose to a human and a disease selected from bacteremia due to bacteria of the genus Egella, sinusitis, purulent myositis, skin abscess, spondylodiscitis and liver abscess Method for.
  8. 　２型糖尿病ならびにエガセラ属細菌による菌血症、副鼻腔炎、化膿性筋炎、皮膚膿瘍、脊椎椎間板炎および肝膿瘍から選択される疾患を患うヒトに、１－ケストースを投与することを含む、前記疾患を治療する方法。 Comprising administering 1-kestose to a human suffering from type 2 diabetes and a disease selected from bacteremia due to Egella spp., sinusitis, purulent myositis, skin abscess, spinal discitis and liver abscess How to treat a disease.
  9. 　エガセラ属細菌の菌数を抑制するための医薬品の製造における１－ケストースの使用。 Use of 1-kestose in the manufacture of pharmaceuticals to control the number of Eggerthella bacteria.
  10. 　２型糖尿病ならびにエガセラ属細菌による菌血症、副鼻腔炎、化膿性筋炎、皮膚膿瘍、脊椎椎間板炎および肝膿瘍から選択される疾患の予防または治療のための医薬品の製造における１－ケストースの使用。 Use of 1-Kestose in the manufacture of pharmaceuticals for the prevention or treatment of types 2 diabetes and diseases selected from Eggerthella-induced bloodstream, sinusitis, pyomyositis, cutaneous abscess, spinal discitis and liver abscess ..
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WO2023284848A1 (en) * 2021-07-15 2023-01-19 量子高科(广东)生物有限公司 Fructooligosaccharide having high kestose content and application thereof

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