WO2025023183A1 - 炎症性腸疾患を抑制する脂肪酸代謝産物 - Google Patents

炎症性腸疾患を抑制する脂肪酸代謝産物 Download PDF

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WO2025023183A1
WO2025023183A1 PCT/JP2024/025989 JP2024025989W WO2025023183A1 WO 2025023183 A1 WO2025023183 A1 WO 2025023183A1 JP 2024025989 W JP2024025989 W JP 2024025989W WO 2025023183 A1 WO2025023183 A1 WO 2025023183A1
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inflammatory bowel
bowel disease
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treating
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French (fr)
Japanese (ja)
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茂呂 和世 大谷
俊哉 畑井
泰隆 本村
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University of Osaka NUC
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    • 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
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
    • A61K31/202—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids having three or more double bonds, e.g. linolenic
    • 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
    • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
    • A61K31/22—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin
    • A61K31/23—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin of acids having a carboxyl group bound to a chain of seven or more carbon atoms
    • A61K31/232—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin of acids having a carboxyl group bound to a chain of seven or more carbon atoms having three or more double bonds, e.g. etretinate
    • 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
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • 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]

Definitions

  • the present invention relates to a pharmaceutical composition for treating and/or preventing inflammatory bowel disease, specifically, the present invention relates to a pharmaceutical composition for treating and/or preventing inflammatory bowel disease, such as ulcerative colitis or Crohn's disease, which contains a group of compounds including 11-hydroxy-eicosapentaenoic acid.
  • Inflammatory bowel disease is an immune disease designated as an intractable disease, with the main symptoms being diarrhea, bloody stools, and abdominal pain. It is broadly divided into two types: ulcerative colitis and Crohn's disease. Currently, the number of ulcerative colitis patients in Japan is estimated to be about 200,000, and Crohn's disease patients to about 45,000, with the number of patients increasing year by year. In ulcerative colitis, chronic inflammation occurs continuously from the large intestine to the rectum, while Crohn's disease is a disease in which chronic inflammation occurs in the entire digestive tract from the mouth to the anus. In both cases, inflammatory bowel disease occurs when immune cells in the intestinal tract react to intestinal bacteria that normally do not cause an immune response, producing inflammatory cytokines. With the development of drugs that neutralize inflammatory cytokines, it has become possible to control the condition to a certain extent.
  • Medicinal drugs used to treat ulcerative colitis include 5-aminosalicylates (e.g., 5-ASA: salazosulfapyridine, mesalazine), corticosteroids (e.g., prednisolone, budesonide), cytapheresis, immunomodulators or suppressants (e.g., azathioprine, 6-mercaptopurine, cyclosporine, tacrolimus), anti-TNF ⁇ antagonists (e.g., infliximab, adalimumab, golimumab), anti-adhesion molecule antibodies (e.g., vedolizumab), anti-interleukin 12/23 antagonists (e.g., ustekinumab), and Janus kinase inhibitors (e.g., tofacitinib) ( https://www.nanbyou.or.jp/entry/62 ).
  • 5-aminosalicylates e.g
  • Non-Patent Document 1 A cohort study in 2001 reported that patients who underwent appendectomy before the age of 20 had a lower lifetime risk of developing ulcerative colitis (Non-Patent Document 1). It has been reported that the appendix is a safe house for intestinal bacteria, and that it is a site where lymphatic tissue accumulates and produces immunoglobulin A (Non-Patent Document 2). However, the mechanism by which appendectomy suppresses colitis is unknown. Therefore, the inventors created appendectomy mice and analyzed the changes that occur in the body, as well as inducing colonic inflammation in these mice and analyzing the effect of appendectomy on the onset of colitis, thereby investigating the mechanism by which appendectomy suppresses the onset of colitis.
  • Non-Patent Document 3 tuft cells (brush cells), which are known as epithelial cells that produce a large amount of IL-25 during parasitic infection and induce the activation of type 2 cytokine-producing cells, and it was revealed that this caused the production of IL-25, the main cytokine produced by tuft cells, to increase, which played an important role in alleviating colitis.
  • brush cells which are known as epithelial cells that produce a large amount of IL-25 during parasitic infection and induce the activation of type 2 cytokine-producing cells, and it was revealed that this caused the production of IL-25, the main cytokine produced by tuft cells, to increase, which played an important role in alleviating colitis.
  • Non-Patent Document 4 On the other hand, appendectomy has been reported to change the colonic flora in both mice and humans.
  • ulcerative colitis carries the risk that the disease can easily develop into colon cancer if it continues for a long time, and that patients with the most severe cases may be forced to have their colon removed. In light of this current situation, there is a need for the development of new treatments based on elucidating the mechanism behind the onset of ulcerative colitis.
  • the inventors focused on the above-mentioned changes in the colonic bacterial flora caused by appendectomy and conducted extensive research into elucidating the factors that induce tuft cell hyperplasia due to changes in the bacterial flora.
  • LCFA long-chain fatty acid
  • 11-HEPE 11-hydroxy-eicosapentaenoic acid
  • composition includes the following aspects.
  • a composition for treating and/or preventing inflammatory bowel disease comprising a compound represented by the following formula (I): [2] The composition according to [1], wherein R1 is hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted aryl.
  • composition according to [3] The composition according to [2], wherein R1 is hydrogen or unsubstituted alkyl having 1 to 4 carbon atoms.
  • R1 is hydrogen or unsubstituted alkyl having 1 to 4 carbon atoms.
  • composition according to any one of [1] to [5], wherein the composition is a pharmaceutical composition.
  • a food composition for treating and/or preventing inflammatory bowel disease comprising Acaryochloris marina and/or Myxococcus xanthus.
  • the food composition according to [15] which is prepared in the form of a tablet, pill, capsule, powder, granule, fine granule, troche, liquid, nasogastric tube feeding agent, or enteral nutrition agent.
  • Figure 1 shows that appendectomy results in a robust mucin layer.
  • Figure 1A is a graph showing the results of q-PCR for muc2 from colons isolated from sham-operated (sham) and appendectomized (APX) mice. * indicates p ⁇ 0.05.
  • Figure 1B shows the results of PAS (Periodic Acid Schiff) staining of colons isolated from sham-operated (sham) and appendectomized (APX) mice.
  • Figure 1C shows the results of FISH (Fluorescence in situ hybridization) staining of colons isolated from sham-operated (sham) and appendectomized (APX) mice. The mucus layer is within the white dotted line.
  • Figure 1D shows the area of the mucus layer in Figure 1C.
  • Figure 2 shows the bacterial counts in feces from sham-operated (sham) and appendectomized (APX) mice.
  • Figure 2A shows the results reported previously, and
  • Figure 2B shows the results of this experiment.
  • 1-6 are the individual mouse numbers.
  • Figure 3 is a schematic diagram showing the single-cage and shared-cage rearing of sham-operated and appendectomized mice. "Single-housed” and “shared-housed” on the left indicate the rearing method, and "sham-operated” and “appendectomized” on the mice indicate the type of surgery they underwent.
  • Figure 4A shows the change in body weight during induction of colitis in sham-operated (sham) and appendectomized (APX) mice housed in single or shared cages.
  • Figure 4B shows the length of the large intestine.
  • Figure 4C shows the amount of inflammatory cytokines IL-1 ⁇ and IL-6 in colon tissue suspensions.
  • Figure 4D shows the results of H&E staining of the large intestine.
  • co-housed sham-operated when sham-operated and appendectomized mice were housed in the same cage, “co-housed sham-operated” refers to the mice in that cage that underwent sham surgery, and “co-housed appendectomy” refers to the mice in that cage that underwent appendectomy.
  • FIG. 5 shows graphs showing changes in body weight (A) and colon length (B) in mice administered four types of antibiotics, ampicillin, vancomycin, metronidazole, and neomycin, and then housed in a cage with sham-operated or appendectomized mice, and then administered dextran sodium sulfate (DSS).
  • FIG. 6 is a graph showing the amount of tuft cells (A) and IL-25 (B) in sham-operated and appendectomized mice housed in single cages, and in sham-operated and appendectomized mice housed in shared cages.
  • Figure 7A shows the results of measuring the peaks of each metabolite by LCMS in sham-operated (sham) or appendectomized (APX) mice.
  • Figure 7B shows a graph showing that 11-HEPE is significant in suppressing weight loss in DSS-induced colitis.
  • Figure 7C shows that 11-HEPE is significant in suppressing colon shortening in DSS-induced colitis.
  • Figure 8 shows the differences in the abundance of bacterial species (A) or fungal species (B) identified by shotgun metagenomic sequencing of feces from sham-operated (sham) or appendectomized (APX) mice.
  • Figure 9 is a graph showing the OTU counts of Acaryochloris marina and Myxococcus xanthus in fecal samples from sham-operated and appendectomized mice housed in single cages, and from sham-operated and appendectomized mice housed in shared cages.
  • the present invention relates to a compound represented by formula (I):
  • R1 is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, or substituted or unsubstituted cycloalkenyl or substituted or unsubstituted heterocyclyl, preferably hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted aryl, more preferably hydrogen or unsubstituted alkyl having 1 to 4 carbon atoms] or a pharma- ceutically acceptable salt thereof, for use in treating and/or preventing inflammatory bowel disease.
  • the present invention is based on the discovery that 11-HEPE acts directly on epithelial cells in colonic organoids to enhance the expression of IL-25.
  • the enhanced production of IL-25 associated with the activation of tuft cells by 11-HEPE activates group 2 innate lymphoid cells (ILC2) and promotes the production of IL-13.
  • IL-13 acts on epithelial stem cells to enhance the formation of mucin-producing goblet cells and tuft cells. Without being bound by any particular theory, it is thought that the thickening of the mucin layer isolates intestinal bacteria from epithelial cells and inhibits inadvertent adhesion, creating a situation in which colitis is less likely to occur.
  • WO2018/150257 JP 2020-508286 discloses the use of eicosapentaenoic acid as a high-purity free fatty acid to reduce inflammation or modify microbial sacs in patients with ulcerative colitis, but does not mention 11-HEPE.
  • WO2007/076531 JP Patent Publication No.
  • a method for preventing or treating inflammatory bowel disease (IBD) in cats which comprises administering a composition containing docosahexaenoic acid (DHA) and, optionally, one or more fatty acids selected from the group consisting of eicosapentaenoic acid (EPA), arachidonic acid (ARA), linoleic acid (LA), and ⁇ -linoleic acid (ALA), but does not disclose 11-HEPE.
  • DHA docosahexaenoic acid
  • ARA arachidonic acid
  • LA linoleic acid
  • ALA ⁇ -linoleic acid
  • WO2021/019037 JP 2022-542513 discloses a method for resolving inflammatory symptoms by significantly and unexpectedly enhancing the biosynthesis of specific inflammation-resolving lipid mediators (SPMs) using a preparation containing a specific extract and ⁇ -3 fatty acid.
  • SPMs inflammation-resolving lipid mediators
  • 11-HEPE is merely exemplified as an example of SPM.
  • JP 2015-163607 A discloses a method for obtaining hydroxyeicosapentaenoic acid (8-HEPE, 9-HEPE, 12-HEPE) by extraction from krill and/or shrimp. It also discloses that 11-HEPE has excellent activation of PPAR ⁇ , PPAR ⁇ , and PPAR ⁇ , and that HEPE with excellent PPAR activation has a hydroxyl group bonded at the center of the carbon chain (8th to 12th positions). Activation of PPAR is expected to have effects such as activation of fatty acid metabolism, promotion of body fat burning, suppression of obesity, suppression of fatty liver, prevention and improvement of diabetes, prevention and improvement of insulin resistance, and improvement of endurance, but there is no description regarding ulcerative colitis. 6.
  • JP 2010-248213 A discloses a method for treating or preventing inflammation, which is characterized by administering 15-hydroxyeicosapentaenoic acid, 18-hydroxyeicosapentaenoic acid, 5,12,18-hydroxyeicosapentaenoic acid, or 5,6,15-hydroxyeicosapentaenoic acid.
  • 11-HEPE discloses a method for treating or preventing gastrointestinal conditions (ulcerative colitis, Crohn's disease, etc.) by administering 5,6,15-hydroxyeicosapentaenoic acid, etc.
  • 11-HEPE discloses a method for treating or preventing inflammation, which is characterized by administering 15-hydroxyeicosapentaenoic acid, 18-hydroxyeicosapentaenoic acid, 5,12,18-hydroxyeicosapentaenoic acid, or 5,6,15-hydroxyeicosapentaenoic acid.
  • WO2007/076531 JP Patent Publication
  • WO2003/076531 JP Patent Publication 2003-525880 discloses a method for treating or preventing inflammation, which is characterized by administering 15-hydroxyeicosapentaenoic acid, 18-hydroxyeicosapentaenoic acid, 5,12,18-hydroxyeicosapentaenoic acid, or 5,6,15-hydroxyeicosapentaenoic acid.
  • 11-HEPE there is no description of 11-HEPE.
  • the present invention provides a composition for treating and/or preventing inflammatory bowel disease, comprising a compound of formula (I) containing 11-HEPE, or a pharma- ceutically acceptable salt thereof.
  • the substituents in formula (I) are defined as follows:
  • alkyl refers to a straight-chain or branched hydrocarbon group having 1 to 10 carbon atoms. It includes alkyl having 1 to 6 carbon atoms, alkyl having 1 to 4 carbon atoms, alkyl having 1 to 3 carbon atoms, etc.
  • Examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, hexyl, isohexyl, n-heptyl, isoheptyl, n-octyl, isooctyl, n-nonyl, n-decyl, etc.
  • Particularly preferred are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, etc.
  • alkenyl refers to a straight-chain or branched hydrocarbon group having 2 to 10 carbon atoms and having one or more double bonds at any position. It includes alkenyl having 2 to 8 carbon atoms and alkenyl having 2 to 6 carbon atoms. Examples include vinyl, propenyl, isopropenyl, butenyl, isobutenyl, prenyl, butadienyl, pentenyl, isopentenyl, pentadienyl, hexenyl, isohexenyl, hexadienyl, heptenyl, octenyl, nonenyl, decenyl, etc. In particular, vinyl, propenyl, isopropenyl, butenyl, isobutenyl, etc. are preferred.
  • alkynyl refers to a linear or branched hydrocarbon group having 2 to 10 carbon atoms and having one or more triple bonds at any position. It includes alkynyl having 2 to 6 carbon atoms, alkynyl having 2 to 4 carbon atoms, etc. Examples include ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl, etc. Alkynyl may have a double bond in addition to one or more triple bonds at any position. In particular, ethynyl, propynyl, butynyl, etc. are preferred.
  • aryl refers to a monocyclic or polycyclic aromatic carbocyclic group, and a group in which one or two 3- to 8-membered rings are further condensed to such a monocyclic or polycyclic aromatic carbocyclic group.
  • monocyclic or polycyclic aromatic carbocyclic groups include phenyl, naphthyl, anthryl, and phenanthryl. Phenyl is particularly preferred.
  • heteroaryl includes 5- to 6-membered aromatic ring groups having at least one nitrogen atom, oxygen atom, and/or sulfur atom in the ring.
  • condensed aromatic heterocyclic groups include benzoxalinyl, purine, pteridinyl, benzimidazolyl, benzisoxazolyl, benzoxazolyl, benzoxadiazolyl, benzisothiazolyl, benzothiazolyl, benzothiadiazolyl, benzofuranyl, isobenzofuranyl, benzothiophenyl, benzotriazolyl, imidazopyridinyl, triazolopyridinyl, imidazothiazolyl, pyrazinopyridazinyl, and benzimidazolinyl.
  • cycloalkyl includes monocyclic or polycyclic saturated carbocyclic groups having 3 to 10 carbon atoms.
  • monocyclic cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononanyl, and cyclodecanyl.
  • polycyclic cycloalkyl include norbornanyl, tetrahydronaphthalenyl, and adamantanyl. Cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl are particularly preferred.
  • cycloalkenyl includes non-aromatic monocyclic or polycyclic ring groups having 3 to 10 carbon atoms and containing at least one carbon-carbon double bond.
  • monocyclic cycloalkenyls include cyclopentenyl and cyclohexenyl.
  • polycyclic cycloalkenyls include norbornenyl and indenyl. In particular, cyclopentenyl, cyclohexenyl, etc. are preferred.
  • heterocyclyl includes a 5- to 7-membered non-aromatic cyclic group having at least one nitrogen atom, oxygen atom and/or sulfur atom in the ring, a non-aromatic cyclic group in which two or more of these are independently fused, a cyclic group in which a 5- to 7-membered aromatic ring having at least one nitrogen atom, oxygen atom and/or sulfur atom in the ring is fused with one or more of the above-mentioned "cycloalkyl” or “cycloalkenyl” ring portions, and a cyclic group in which a 5- to 7-membered non-aromatic heterocyclic group having at least one nitrogen atom, oxygen atom and/or sulfur atom in the ring is fused with one or more of the above-mentioned "aryl” or “heteroaryl” ring portions.
  • heterocyclic group examples include monocyclic non-aromatic heterocyclic groups such as pyrrolinyl, pyrrolidinyl, imidazolinyl, imidazolidinyl, pyrazolinyl, pyrazolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, dihydropyridinyl, dihydropyridazinyl, dihydropyrazinyl, dioxanyl, oxathiolanyl, thianyl, tetrahydrofuranyl, tetrahydrothiazolinyl, and tetrahydroisothiazolinyl, and condensed non-aromatic heteroaromatic groups such as indolinyl, isoindolinyl, benzopyranyl, benzodioxanyl, tetrahydroquinolinyl, and te
  • substituents of "substituted alkyl”, “substituted alkenyl”, and “substituted alkynyl” include halogen, hydroxy, mercapto, nitro, nitroso, cyano, etc. Halogen is particularly preferred.
  • substituents of "substituted aryl", “substituted heteroaryl”, “substituted cycloalkyl”, “substituted cycloalkenyl” and “substituted heterocyclyl” are preferably halogen, cyano, hydroxy, formyl, carboxy, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted alkyloxy, substituted or unsubstituted alkenyloxy, substituted or unsubstituted alkynyloxy, etc.
  • halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkyloxy, etc. are preferred.
  • the compound represented by formula (I) exists in the S-form and the R-form, and in the present invention, the compound represented by formula (I) includes each of the optically active and racemic forms of the S-form and the R-form, as well as mixtures in which the S-form and the R-form are present in any amount.
  • the present invention provides a composition for treating and/or preventing inflammatory bowel disease, comprising a pharma- ceutically acceptable salt of a compound represented by formula (I).
  • pharma- ceutically acceptable salts of the compound represented by formula (I) include salts of the compound represented by formula (I) with alkali metals (e.g., lithium, sodium, potassium, etc.), alkaline earth metals (e.g., calcium, barium, etc.), magnesium, transition metals (e.g., zinc, iron, etc.), ammonia, organic bases (e.g., trimethylamine, triethylamine, dicyclohexylamine, ethanolamine, diethanolamine, triethanolamine, meglumine, ethylenediamine, pyridine, picoline, quinoline, etc.) and amino acids, or salts of inorganic acids (e.g., hydrochloric acid, sulfuric acid, nitric acid, carbonic acid, hydrobromic
  • inorganic acids e.
  • Prodrugs The compound of formula (I) or a pharma- ceutically acceptable salt thereof may form a prodrug.
  • the present invention provides a composition for treating and/or preventing inflammatory bowel disease, comprising a prodrug of the compound of formula (I).
  • a prodrug is a derivative of the compound of formula (I) having a chemically or metabolically decomposable group, which is converted to a pharma- ceutically active compound in vivo by solvolysis or under physiological conditions.
  • Prodrugs include compounds that are converted to the compound of formula (I) by enzymatic oxidation, reduction, hydrolysis, etc.
  • examples of prodrugs include acyloxy derivatives and sulfonyloxy derivatives, which are produced by reacting the compound represented by formula (I) with an appropriate acyl halide, an appropriate acid anhydride, an appropriate sulfonyl chloride, an appropriate sulfonyl anhydride and a mixed anhydride, or by reacting the compound represented by formula (I) using a condensing agent.
  • the prodrug of the compound represented by formula (I) may be the above-mentioned prodrug, or may form a triglyceride, diglyceride, or monoglyceride in which one, two, or three of the three hydroxyl groups of glycerin are esterified with the compound represented by formula (I) and/or other fatty acid.
  • a triglyceride in which all three hydroxyl groups of glycerin are esterified with the compound represented by formula (I) a triglyceride in which two compounds represented by formula (I) and other fatty acid are esterified, or a triglyceride in which one compound represented by formula (I) and two other fatty acid are esterified may be formed.
  • a diglyceride a diglyceride in which all two hydroxyl groups of glycerin are esterified with the compound represented by formula (I), or a diglyceride in which one compound represented by formula (I) and other fatty acid are esterified may be formed.
  • compositions of the present invention can be pharmaceutical compositions and food compositions.
  • the pharmaceutical composition of the present invention is a composition for treating and/or preventing inflammatory bowel disease.
  • the food composition of the present invention may be a composition for treating and/or preventing inflammatory bowel disease, or may be a composition for treating and/or preventing diarrhea, bloody stool and/or abdominal pain, particularly diarrhea, bloody stool and/or abdominal pain that are thought to be caused by inflammatory bowel disease.
  • the present invention relates to a pharmaceutical composition for treating and/or preventing inflammatory bowel disease, which comprises a compound represented by formula (I) or a pharma- ceutically acceptable salt thereof.
  • the present invention relates to a pharmaceutical composition for treating and/or preventing inflammatory bowel disease, comprising linoleate 11-lipoxygenase (11-LOX).
  • 11-LOX 11-lipoxygenase
  • eicosapentaenoic acid is enzymatically converted by linoleic acid-11-lipoxygenase (11-LOX) to produce 11-hydroxy-eicosapentaenoic acid (11-HEPE).
  • compounds represented by formula (I) such as 11-HEPE, can treat and/or prevent inflammatory bowel disease. Therefore, it is believed that 11-LOX, which can induce 11-HEPE, can also treat or manage and/or prevent inflammatory bowel disease.
  • the present invention relates to a pharmaceutical composition for treating and/or preventing inflammatory bowel disease, comprising Acaryochloris marina and/or Myxococcus xanthus.
  • a pharmaceutical composition for treating and/or preventing inflammatory bowel disease comprising Acaryochloris marina and/or Myxococcus xanthus.
  • 11-LOX is also believed to be capable of treating and/or preventing inflammatory bowel disease
  • the present inventors have searched for the origin of 11-LOX and found that it is derived from Acaryochloris marina or Myxococcus xanthus. Since 11-LOX can treat and/or prevent inflammatory bowel disease in the present invention, it is believed that Acaryochloris marina or Myxococcus xanthus, which are its origins, can also treat and/or prevent inflammatory bowel disease.
  • the invention relates in another embodiment to Acaryochloris marina and/or Myxococcus xanthus as probiotics for treating and/or preventing inflammatory bowel disease.
  • Probiotics means live microorganisms which, when taken in sufficient amounts, confer a beneficial effect on the host.
  • the present invention relates in a further embodiment to a microbial pharmaceutical composition for treating and/or preventing inflammatory bowel disease, comprising Acaryochloris marina and/or Myxococcus xanthus.
  • the present invention relates to fecal microbiota transplantation (FMT) using a microbiota containing Acaryochloris marina and/or Myxococcus xanthus.
  • Fecal transplantation is generally a treatment in which intestinal bacteria contained in the stool of a healthy person are administered to a sick patient, and is also known as gut microbiota transplantation.
  • intestinal bacteria contained in the stool of a healthy person are administered to a sick patient, and is also known as gut microbiota transplantation.
  • gut microbiota transplantation for example, inflammatory bowel disease can be treated and/or prevented by selecting a donor by confirming that the donor's stool contains Acaryochloris marina and/or Myxococcus xanthus, and administering the microbiota obtained from the donor to the patient.
  • the present invention relates to a bacterial flora comprising Acaryochloris marina and/or Myxococcus xanthus for treating or preventing inflammatory bowel disease.
  • the present invention relates to the use of a bacterial flora comprising Acaryochloris marina and/or Myxococcus xanthus for the manufacture of a medicament for treating or preventing inflammatory bowel disease.
  • treatment means a method or process that aims to (1) delay the onset of inflammatory bowel disease; (2) slow or stop the progression, worsening or aggravation of the symptoms of inflammatory bowel disease; (3) bring about the amelioration of the symptoms of inflammatory bowel disease; or (4) cure inflammatory bowel disease.
  • Treatment may be administered prior to the onset of the disease or condition as a preventative measure, or treatment may be administered after the onset of the disease.
  • inflammatory bowel disease means an immune disease designated as an intractable disease whose main symptoms are diarrhea, bloody stools, and abdominal pain, and includes ulcerative colitis and Crohn's disease. Ulcerative colitis is a disease in which chronic inflammation occurs continuously from the large intestine to the rectum, while Crohn's disease is a disease in which chronic inflammation occurs in the entire digestive tract from the mouth to the anus.
  • prevention means preventing the onset of inflammatory bowel disease in advance.
  • compositions of the present invention can be administered by a variety of conventional routes of administration, including orally, topically, parenterally, e.g., intravenously or subcutaneously. Additionally, the compositions of the present invention can be administered intranasally or as a rectal suppository.
  • compositions of the present invention may be administered in single or multiple doses, with the active ingredient alone or in combination with a pharma- ceutically acceptable carrier, vehicle or diluent.
  • Suitable pharmaceutical carriers, vehicles and diluents include inert solid diluents or fillers, sterile aqueous solutions and various organic solvents.
  • the pharmaceutical compositions formed by combining the composition with a pharma- ceutically acceptable carrier, vehicle or diluent are easy to administer in a variety of dosage forms, such as tablets, powders, lozenges, syrups, injectable solutions, and the like.
  • These pharmaceutical compositions may contain additional ingredients, such as flavorings, binders, excipients, and the like, if desired.
  • solutions of the compositions may be prepared, for example, in sesame or peanut oil, aqueous propylene glycol, or may be used in sterile aqueous solutions.
  • aqueous solutions should be suitably buffered if necessary and sufficient saline or glucose first rendered isotonic.
  • These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration.
  • compositions of the present invention can be administered to humans and animals in unit dosage forms, such as tablets, capsules, pills, powders, granules, suppositories, sterile parenteral solutions or suspensions, sterile parenteral solutions of suspensions, and oral solutions or suspensions, containing an appropriate amount of the active ingredient.
  • compositions of the present invention are mixed with one or more pharma- ceutically acceptable carriers and/or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and/or silicic acid; (2) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and/or gum arabic; (3) humectants, such as glycerol; (4) disintegrants, such as powdered agar, calcium carbonate, etc.
  • fillers or extenders such as starch, lactose, sucrose, glucose, mannitol, and/or silicic acid
  • binders such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and/or gum arabic
  • humectants such as glycerol
  • disintegrants such as powdered agar, calcium carbonate, etc.
  • poor solvents such as paraffin
  • absorption enhancers such as quaternary ammonium compounds
  • wetting agents such as acetyl alcohol and glycerol monostearate
  • absorbents such as kaolin and bentonite clay
  • lubricants such as talc, calcium stearate, magnesium stearate, polyethylene glycol solids, sodium lauryl sulfate, and mixtures thereof
  • coloring agents such as paraffin, (6) absorption enhancers, such as quaternary ammonium compounds; (7) wetting agents, such as acetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such as talc, calcium stearate, magnesium stearate, polyethylene glycol solids, sodium lauryl sulfate, and mixtures thereof; and (10) coloring agents.
  • Liquid dosage forms for oral administration include pharma- ceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs.
  • liquid dosage forms can contain inert diluents commonly used in the art, such as water or other solvents, solubilizing and emulsifying agents, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (especially cottonseed oil, corn oil, peanut oil, sunflower oil, soybean oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.
  • inert diluents commonly used in the art, such as water or other solvents, solubilizing and emuls
  • the suspension may contain suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol, and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, powdered agar, and tragacanth, and mixtures thereof.
  • suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol, and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, powdered agar, and tragacanth, and mixtures thereof.
  • compositions of the present invention can be administered as compositions for rectal administration, for example, as suppositories.
  • Suppositories can be prepared by mixing with one or more suitable non-irritating carriers, including, for example, cocoa butter, polyethylene glycol, suppository wax, or salicylic acid, which are solid at room temperature but liquid at body temperature and melt in a suitable body cavity to release the encapsulated compound(s) and composition(s).
  • suitable non-irritating carriers including, for example, cocoa butter, polyethylene glycol, suppository wax, or salicylic acid, which are solid at room temperature but liquid at body temperature and melt in a suitable body cavity to release the encapsulated compound(s) and composition(s).
  • a composition for rectal administration may be in the form of a liquid dosage form, such as a pharma- ceutically acceptable emulsion, microemulsion, solution, or suspension.
  • the liquid dosage form may contain an inert diluent commonly used in the art, such as water or other solvent, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (especially cottonseed oil, corn oil, peanut oil, sunflower oil, soybean oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan, and mixtures thereof.
  • a prodrug technique may be used to convert an orally administered compound
  • the present invention relates to a method for treating or preventing inflammatory bowel disease, which comprises administering a compound represented by formula (I) of the present invention or a pharma- ceutically acceptable salt thereof to a subject in need of such treatment.
  • the present invention relates to a compound represented by formula (I) of the present invention or a pharma- ceutically acceptable salt thereof for treating or preventing inflammatory bowel disease.
  • the present invention relates to use of a compound represented by formula (I) of the present invention, or a pharma- ceutically acceptable salt thereof, for the manufacture of a medicament for treating or preventing inflammatory bowel disease.
  • the present invention relates to a method for treating or preventing inflammatory bowel disease, comprising administering 11-HEPE to a subject in need of such treatment. Furthermore, in another embodiment, the present invention relates to 11-HEPE for treating or preventing inflammatory bowel disease. In yet another aspect, the present invention relates to use of 11-HEPE for the manufacture of a medicament for treating or preventing inflammatory bowel disease.
  • the present invention relates to a method for treating or preventing inflammatory bowel disease, comprising administering Acaryochloris marina and/or Myxococcus xanthus salts to a subject in need of such treatment. Furthermore, in another aspect, the present invention relates to Acaryochloris marina and/or Myxococcus xanthus for treating or preventing inflammatory bowel disease. In yet another aspect, the present invention relates to the use of Acaryochloris marina and/or Myxococcus xanthus for the manufacture of a medicament for treating or preventing inflammatory bowel disease.
  • the present invention relates to a food composition for treating and/or preventing inflammatory bowel disease, or a food composition for treating and/or preventing diarrhea, bloody stool and/or abdominal pain, particularly diarrhea, bloody stool and/or abdominal pain thought to be caused by inflammatory bowel disease, which contains a compound represented by formula (I) or a pharma- ceutically acceptable salt thereof.
  • treatment refers to a method or process intended to (1) delay the onset of diarrhea, bloody stool and/or abdominal pain; (2) slow or stop the progression, worsening or aggravation of the symptoms of diarrhea, bloody stool and/or abdominal pain; or (3) bring about the amelioration of the symptoms of diarrhea, bloody stool and/or abdominal pain.
  • Treatment may be administered prior to the onset of a disease or condition as a preventative measure, or treatment may be administered after the onset of the disease.
  • prevention means preventing the onset of diarrhea, bloody stool and/or abdominal pain in advance.
  • the food composition of the present invention may consist of only the compound represented by formula (I) or a pharma- ceutically acceptable salt thereof, may consist of only a prodrug of the compound represented by formula (I), may consist of only a monoglyceride, diglyceride or triglyceride of the compound represented by formula (I), or may be incorporated into food (food additives, food, beverages, etc.).
  • the food composition of the present invention may contain the compound represented by formula (I) or the like and may be orally ingested by the subject, and there is no particular limit to the type or shape of the food composition.
  • the present invention relates to a food composition for treating and/or preventing inflammatory bowel disease, comprising linoleate 11-lipoxygenase (11-LOX), or a food composition for treating and/or preventing diarrhea, bloody stool and/or abdominal pain, particularly diarrhea, bloody stool and/or abdominal pain thought to be caused by inflammatory bowel disease.
  • the food composition of this embodiment may further comprise eicosapentaenoic acid or may be administered with a meal.
  • 11-LOX linoleic acid-11-lipoxygenase
  • the food composition of the present invention may consist of only 11-HEPE, or may be blended with food (food additives, food, beverages, etc.).
  • the food composition of the present invention may contain 11-HEPE and may be orally ingested by a subject, and there are no particular limitations on the type or shape of the food composition.
  • the present invention relates to a food composition for treating and/or preventing inflammatory bowel disease, comprising Acaryochloris marina and/or Myxococcus xanthus, or a food composition for treating and/or preventing diarrhea, bloody stools and/or abdominal pain, in particular diarrhea, bloody stools and/or abdominal pain thought to be caused by inflammatory bowel disease.
  • the food composition of this embodiment may further comprise eicosapentaenoic acid or may be administered with a meal.
  • 11-LOX is also believed to be capable of treating and/or preventing inflammatory bowel disease
  • the present inventors have searched for the origin of 11-LOX and found that it is derived from Acaryochloris marina or Myxococcus xanthus. Since 11-LOX can treat and/or prevent inflammatory bowel disease in the present invention, it is believed that Acaryochloris marina or Myxococcus xanthus, from which it originates, can also treat and/or prevent inflammatory bowel disease.
  • the food composition of the present invention may consist solely of Acaryochloris marina and/or Myxococcus xanthus, or may be blended with food (food additives, food, beverages, etc.).
  • the food composition of the present invention may be any food composition that contains Acaryochloris marina or Myxococcus xanthus and can be orally ingested by the subject, with no particular limitations on the type, shape, etc. of the food composition.
  • the invention relates, in another embodiment, to Acaryochloris marina and/or Myxococcus xanthus as probiotics for the treatment and/or prevention of diarrhea, bloody stools and/or abdominal pain, in particular diarrhea, bloody stools and/or abdominal pain which may be due to inflammatory bowel disease.
  • probiotics live microorganisms which, when taken in sufficient amounts, confer a beneficial effect on the host.
  • one aspect of the present invention relates to a food composition.
  • the food composition of the present invention include confectioneries such as troches, drops, candies, ramune, gummies, and chewing gum; Western confectioneries such as cookies, crackers, biscuits, potato chips, bread, cakes, chocolates, donuts, puddings, and jellies; Japanese confectioneries such as rice crackers, yokan, daifuku, ohagi, manju, and castella; frozen desserts such as ice cream, popsicles, sorbets, and gelato; breads such as white bread, French bread, and croissants; noodles such as udon, soba, Chinese noodles, and kishimen; fish paste products such as kamaboko, chikuwa, and fish sausage; meat products such as ham, sausage, hamburger steak, and corned beef; salt, pepper, miso, soy sauce, sauces, dressings, mayonnaise, ketchup, and sweeteners (
  • condiments such as troches
  • Beverages include food and beverages such as soup and miso soup; powdered food and beverages such as instant coffee, instant tea, instant milk, instant soup, and instant miso soup; alcoholic beverages such as whiskey, bourbon, spirits, liqueur, wine, fruit wine, Japanese sake, Chinese wine, shochu, beer, non-alcoholic beer with an alcohol content of 1% or less, sparkling wine, and chuhai; beverages containing fruit juice (e.g., apple, mandarin orange, grape, banana, pear, plum juice, etc.), and vegetable juice (e.g., These include non-alcoholic beverages such as beverages containing vegetable juices (such as tomato, carrot, celery, cucumber, and watermelon), beverages containing fruit and vegetable juices, soft drinks, milk, soy milk, dairy beverages, drink-type yogurt, coffee, cocoa, tea beverages (black tea, green tea, barley tea, brown rice tea, sencha, gyokuro tea, roasted green tea, oolong tea, turmeric tea, pu-erh tea, r
  • Suitable examples of such food compositions include foods or beverages such as natto, yogurt, vegetable oil, jelly, tea drinks, alcoholic drinks, drops, candy, Ramune, cookies, crackers, biscuits, chocolate, cheese, butter, margarine, chewing gum, etc., to which a compound represented by formula (I) or the like has been added.
  • the food composition of the present invention may also be prepared as a functional food, health food, food for specified health uses, food for special dietary uses (e.g., food for sick people such as hospital food, food for sick people, and nursing care food), supplement, etc., and is preferably prepared as a food for specified health uses, food for special dietary uses, or supplement.
  • food for specified health uses e.g., food for sick people such as hospital food, food for sick people, and nursing care food
  • supplement e.g., a food for specified health uses, food for special dietary uses, or supplement.
  • the food composition of the present invention may be in the form of, for example, tablets, pills, capsules (including hard capsules, soft capsules, and microcapsules), powders, granules, fine granules, troches, liquids (including syrups, emulsions, and suspensions), nasogastric tube feeding preparations, and enteral nutrition preparations, with tablets or capsules being preferred.
  • Another preferred embodiment of the food composition of the present invention is a food or beverage to which a compound represented by formula (I) or the like has been added.
  • the food composition of the present invention is preferably a food for specified health uses, a food for special dietary uses, or a supplement, particularly in the form of a tablet or capsule.
  • supplements do not only mean dietary supplements and functional foods for supplementing nutrients, etc., but also health supplements and functional foods that have functions that are useful for maintaining, restoring, improving, etc., health.
  • the food composition of the present invention can be produced, for example, by adding the compound represented by formula (I) or the like by a known method.
  • a tablet food composition can be produced by adding and mixing the compound represented by formula (I) or the like and excipients (e.g., lactose, white sugar, mannitol, corn starch, etc.), sweeteners, flavorings, and other materials, and molding them into a tablet shape by applying pressure with a tablet press or the like.
  • excipients e.g., lactose, white sugar, mannitol, corn starch, etc.
  • sweeteners e.g., lactose, white sugar, mannitol, corn starch, etc.
  • sweeteners e.g., lactose, white sugar, mannitol, corn starch, etc.
  • sweeteners e.g., lactose, white sugar, mannitol, corn starch, etc.
  • sweeteners e
  • the food composition of the present invention may contain physiologically acceptable carriers in addition to commonly used food ingredients, food additives, various nutrients, vitamins, flavoring substances (e.g., cheese, chocolate, etc.), so long as the effects of the present invention are not inhibited.
  • physiologically acceptable carriers include various conventional organic or inorganic carrier substances, such as excipients, binders, disintegrants, lubricants, colorants, sweeteners, preservatives, antioxidants, thickeners, emulsifiers, etc.
  • Food additives include colorants, sweeteners, preservatives, antioxidants, flavoring agents, etc.
  • other materials such as minerals such as iron and calcium, and dietary fibers such as pectin, carrageenan, and mannan, may also be contained.
  • excipients may be the same as those used in the pharmaceutical composition of the present invention described above.
  • Vitamins may be either water-soluble or fat-soluble, and examples include retinol palmitate, ⁇ -tocopherol (vitamin E), bisbentiamine, riboflavin, pyridoxine hydrochloride, cyanocobalamin, sodium ascorbate (vitamin C), cholecalciferol, nicotinamide, calcium pantothenate, folic acid, biotin, choline bitartrate, niacin, and nicotinamide mononucleotide (NMN).
  • Food compositions in the form of tablets, granules, or fine granules may be coated with a coating substrate by a method known per se for purposes such as taste masking, improving photostability, improving appearance, or enteric properties.
  • a coating substrate include the same ones as those used for the pharmaceutical composition of the present invention described above, and coating can be carried out in the same manner.
  • the content of the compound represented by formula (I) in the food composition of the present invention is not particularly limited, and is, for example, 0.1% by weight or more and 50% by weight or less based on the entire food composition.
  • the lower limit is preferably 0.5% by weight or more, and more preferably 1.0% by weight or more, based on the entire food composition.
  • the upper limit is preferably 20% by weight or less, and more preferably 10% by weight or less, based on the entire food composition.
  • the food composition obtained in this manner is safe and can be given continuously to subjects, particularly preferably humans.
  • Example 1 Appendectomy strengthens the mucin layer
  • Example 1-1 Quantitative PCR The colon was removed from sham-operated (sham) or appendectomized (APX) mice. Each colon was cut open longitudinally and the contents were washed with PBS. The colon was cut into 2 mm cubes, and 15 ml of pre-warmed EDTA buffer (5 mM EDTA/PBS) was placed in an Erlenmeyer flask and stirred at 37°C for 30 minutes. The supernatant was collected by passing through a 100 ⁇ m filter, and the pellet was collected by centrifugation at 1500 rpm for 5 minutes. RNAIso was added to the resulting pellet and dissolved well.
  • RNAiso 0.2 times the amount of chloroform as RNAiso was added, mixed by shaking vigorously, and left at room temperature for 5 minutes. Centrifuged at 12000xG for 15 minutes at 4°C. The supernatant was transferred to a new tube, the same amount of isopropanol was added, mixed, and left at room temperature for 10 minutes. Centrifuged at 12000xG for 10 minutes at 4°C. Washed with 75% ethanol. Centrifuged at 7500xG for 5 minutes at 4°C. The supernatant was discarded, leaving the precipitate, and the mixture was dried. The mixture was dissolved in water and RNA was recovered. The RNA was reverse transcribed to prepare cDNA. q-PCR was performed using the cDNA.
  • ⁇ -actin was simultaneously measured as a housekeeping gene (normalizer), and the ⁇ CT value of mucin-2 (mcu2) gene expression, a component of secretory mucus in the small intestine, was calculated.
  • the results are shown in Figure 1A. Appendectomy (APX) was confirmed to significantly increase muc2 gene expression in the large intestine, supporting the strengthening of the mucin layer.
  • Example 1-2 Periodic Acid Schiff Staining Colons were removed from sham-operated (sham) or appendectomized (APX) mice and fixed overnight in 10% formalin solution. They were dehydrated with three phases of methanol, infiltrated and embedded in two phases of xylene and paraffin to prepare paraffin blocks. Sections were prepared at a thickness of 4 mm. Slides were immersed in 0.5% periodic acid for 5 minutes. They were immersed in Schiff's test solution for 15 minutes. They were rinsed thoroughly with running water. They were immersed in hematoxylin for 1 minute. They were rinsed thoroughly with running water. They were dehydrated with three phases of ethanol, dialyzed with three phases of xylene, mounted, and observed under a microscope. The results are shown in Figure 1B. By performing appendectomy (APX), the area of the mucus that was stained dark black was confirmed to have expanded.
  • APX appendectomized
  • Example 1-3 FISH (fluorescence in situ hybridization) staining Colons were excised from sham-operated (sham) or appendectomized (APX) mice. They were immersed in Carnoy's solution and fixed at room temperature for 2 hours. Paraffin blocks were prepared in the same manner as PAS staining, and sections were prepared at a thickness of 4 mm. Fluorescent staining was performed using DAPI antibody for nuclear staining, MUC2 antibody, and EUB338 antibody for intestinal bacteria staining, and the sections were observed under a fluorescent microscope. The results are shown in Figure 1C. The area within the white dotted line is the mucus layer. It can be seen that the mucus layer expanded due to appendectomy. Figure 1D shows the area of the mucus layer in Figure 1C. These confirmed that appendectomy (APX) increases the area of the mucus layer, increasing the distance between epithelial cells and bacteria.
  • APX appendectomized
  • Example 2 Alterations in gut microbiota following appendectomy
  • 251 bp paired-end sequencing of the V1-V2 variable region of the 16S rRNA gene was performed on an Illumina MiSeq platform (Illumina, San Diego, CA, USA).
  • the resulting paired-end sequences were merged, filtered, and denoised using DADA2. Taxonomic assignment was performed using the QIIME2 feature-classifier plugin with the Greengenes 13_8 database. The results are shown in Figure 2.
  • appendectomy APX was found to result in a decrease in Bacteroidia and an increase in Clostridia in the feces.
  • Example 3 Suppression of colitis dependent on bacterial flora changed by appendectomy (APX)
  • Example 3-1 Shared cage rearing experiment As shown in Figure 3, a sham-operated mouse group and an appendectomized mouse group were reared in the same cage for two weeks from the day of surgery, allowing them to share the bacterial flora. The single-rearing groups were reared only with each surgical group.
  • Example 3-2 Rate of change in body weight As shown in the schematic diagram of Figure 3, cages for sham-operated mice only, appendectomy mice only, and cages for sham-operated and appendectomized mice were prepared. After two weeks of postoperative rearing under the above conditions, the rate of change in body weight when colitis was induced with DSS was shown. The results are shown in Figure 4 A. Co-rearing prevented the suppression of weight loss caused by appendectomy.
  • Example 3-3 Shortening of the large intestine The length of the large intestine under the above conditions was examined. The results are shown in Figure 4B. By co-rearing, shortening of the large intestine due to appendectomy was no longer observed.
  • Example 3-4 Inflammatory Cytokines
  • the large intestine was removed from sham-operated (sham) or appendectomized (APX) mice.
  • the large intestine was cut open longitudinally, and the contents were washed with PBS.
  • the large intestine was cut into 2 mm cubes, and 15 ml of pre-warmed EDTA buffer (5 mM EDTA/PBS) was placed in an Erlenmeyer flask and stirred at 37°C for 30 minutes.
  • the supernatant was collected by passing through a 100 ⁇ m filter, and the pellet was collected by centrifugation at 1500 rpm for 5 minutes.
  • the collected pellet was dissolved in cell lysis buffer, centrifuged at 1500 rpm for 5 minutes, and the supernatant was collected (epithelial fraction).
  • the large intestine from which the epithelium had been removed was homogenized in cell lysis buffer, and the supernatant was collected by centrifugation at 1500 rpm for 5 minutes (lamina propria fraction).
  • the amounts of IL-1 ⁇ , IL-6, and TNF- ⁇ in the lamina intestinal fraction were measured by ELISA.
  • the total protein amount was measured by the Bradford method and normalized. The results are shown in Figure 4C.
  • the inflammatory cytokines IL-1 ⁇ and IL-6 which were decreased by appendectomy, were no longer observed in the co-rearing group. Inflammation occurred at the same level as in the sham operation.
  • Example 3-5 H&E staining Slides were prepared according to the method of Example 1-2. The resulting slides were immersed in hematoxylin solution for 30 seconds, washed thoroughly with running water, and then immersed in eosin solution for 1 minute. They were dehydrated, dialyzed, mounted, and observed under a microscope. The results are shown in Figure 4D. The disappearance of crypts, muscularis hypertrophy, and inflammatory cell infiltration, which are indicators of the severity of colitis, were suppressed by appendectomy, but these suppressions were no longer observed when the mice were kept in the same cage.
  • mice were given free access to water containing four antibiotics, ampicillin, vancomycin, metronidazole, and neomycin, for two weeks. These mice were housed in cages with sham-operated or appendectomized mice for two weeks (drinking normal water). After the two-week period, the mice that had previously been given antibiotics were administered dextran sulfate sodium (DSS) to induce colitis. Body weights were measured for eight days after the start of DSS drinking, and colon lengths were measured eight days after the start of DSS drinking. The results are shown in Figures 5A and B. Compared to the group housed with sham-operated mice, weight loss and shortening of the large intestine were significantly suppressed in the group housed with appendectomized mice.
  • DSS dextran sulfate sodium
  • Example 5 Tuft cell hyperplasia depends on changes in bacterial flora
  • Example 5-1 Tuft cell count The colon was removed from sham-operated (sham) or appendectomized (APX) mice. The colon was cut open longitudinally and the contents were washed with PBS. The colon was cut into 2 mm cubes, and 15 ml of pre-warmed EDTA buffer (5 mM EDTA/PBS) was placed in an Erlenmeyer flask and stirred at 37°C for 30 minutes. The supernatant was collected by passing through a 100 ⁇ m filter, and the pellet was collected by centrifugation at 1500 rpm for 5 minutes.
  • pre-warmed EDTA buffer 5 mM EDTA/PBS
  • the collected pellet was stained with anti-CD45.2, anti-CD326, anti-CD24, and anti-Siglec-F.
  • CD45.2-negative, CD326-positive, CD24-positive, and Siglec-F-positive cells were identified as tuft cells by flow cytometry, and the cell numbers were counted. The results are shown in Figure 6A. The increase in colonic tuft cells following appendectomy was not observed when the mice were reared in sham-operated mice.
  • Example 5-2 IL-25 protein amount
  • Large intestines were excised from sham-operated (sham) or appendectomized (APX) mice.
  • the large intestines were cut open longitudinally, and the contents were washed with PBS.
  • the large intestines were cut into 2 mm cubes, and 15 ml of pre-warmed EDTA buffer (5 mM EDTA/PBS) was placed in an Erlenmeyer flask and stirred at 37°C for 30 minutes. The supernatant was collected while passing through a 100 ⁇ m filter, and the pellet was collected by centrifugation at 1500 rpm for 5 minutes.
  • pre-warmed EDTA buffer 5 mM EDTA/PBS
  • the collected pellet was dissolved in cell lysis buffer, centrifuged at 1500 rpm for 5 minutes, and the supernatant was collected (epithelial fraction).
  • the large intestines from which the epithelium had been removed were homogenized in cell lysis buffer, and the supernatant was collected by centrifugation at 1500 rpm for 5 minutes (lamina propria fraction).
  • the amount of IL-25 in the epithelial fraction was measured by ELISA. Total protein amount was measured by the Bradford method and normalized. The results are shown in Figure 6B. The increase in colonic IL-25 following appendectomy was not observed when the mice were reared in the same compartment as sham-operated mice.
  • Example 6-1 Identification of Metabolites The peaks of each metabolite were identified by LCMS, and metabolites that were significantly increased were selected. Specifically, methanol was added to the feces, and the feces were disrupted and extracted using zirconia beads (6500 rpm, 15 seconds x 2). Extraction was performed overnight at -30°C. The obtained sample was centrifuged (1600g x 10 minutes, 4°C), 200 ⁇ l of the supernatant was transferred to a new tube, 200 ⁇ l of ultrapure water was added, and the mixture was vortexed and centrifuged (10000g x 1 minute, 4°C).
  • Example 6-2 Effect of 11-HEPE Mice were put to sleep under anesthesia 7, 5, 3, and 1 day before the start of DSS drinking, and 11-HEPE was administered enterally. Body weight was measured from the start of DSS drinking until the 8th day, and the rate of change in body weight was measured. In addition, the length of the large intestine on the 8th day was measured. Administration of 11-HEPE significantly suppressed weight loss (Figure 7B) and colon shortening (Figure 7C).
  • Example 7 Identification of long-chain fatty acids that improve colitis through tuft cell hyperplasia It has been reported that eicosapentaenoic acid contained in food is enzymatically converted by linoleate 11-lipoxygenase (11-LOX) to produce 11-hydroxy-eicosapentaenoic acid (11-HEPE) (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6210903/pdf/jlrM088823.pdf, Stabilization and improved activity of arachidonate 11S-lipoxygenase from proteobacterium Myxococcus xanthus).
  • 11-LOX 11-lipoxygenase
  • 11-HEPE 11-hydroxy-eicosapentaenoic acid
  • 11-LOX originates from, i.e., whether it is present in the host or in intestinal bacteria.
  • BRENDA https://www.brenda-enzymes.org/
  • 11-LOX is also known as linoleic acid-11-lipoxygenase and is registered under the EC number 1.13.11.45.
  • the sources of this enzyme are registered as bacteria and fungi, but not animals, especially mammals.
  • Figure 8 shows that appendectomy caused significant changes in the bacterial flora, with an increase in various bacterial species ( Figure 8A), whereas there was little change in the fungal flora, and no fungal species that increased following appendectomy were identified ( Figure 8B). Therefore, we concluded that bacteria, rather than fungi, are the major source of 11-LOX production.
  • Example 8 Changes in Acaryochloris marina and Myxococcus xanthus in feces due to appendectomy
  • a joint cage rearing experiment was carried out according to the method described in Example 3. Cages containing only sham-operated mice, cages containing only appendectomized mice, and cages containing both sham-operated and appendectomized mice were prepared, and the OTU counts of Acaryochloris marina and Myxococcus xanthus were measured in mouse fecal samples from each cage.
  • mice were collected from four groups of mice, namely, mice reared alone with sham surgery, mice reared alone with appendectomy, mice reared together with sham surgery, and mice reared together with appendectomy, and shotgun metagenomic analysis was performed to identify the bacterial species.
  • the colitis-suppressing effect was observed in the "appendectomy alone” group, and as shown in Example 3, the colitis-suppressing effect disappeared in the "appendectomy together with appendectomy” group, despite the mice being subjected to appendectomy.
  • composition includes the following aspects.
  • a composition for treating and/or preventing inflammatory bowel disease comprising a compound represented by the following formula (I): [2] The composition according to [1], wherein R1 is hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted aryl.
  • a food composition for treating and/or preventing inflammatory bowel disease comprising Acaryochloris marina and/or Myxococcus xanthus.
  • the food composition according to [15] which is prepared in the form of a tablet, pill, capsule, powder, granule, fine granule, troche, liquid, nasogastric tube feeding agent, or enteral nutrition agent.

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