Drugs and foods for treating and / or preventing enteritis and / or hepatitis

Abstract

The present invention provides a drug for treating and / or preventing enteritis and / or hepatitis, said drug containing D-amino acid as an active ingredient. In addition, the present invention provides a food composition for suppressing enteritis and / or hepatitis, said composition containing D-amino acid as an active ingredient. In addition, the present invention provides a method for assisting diagnosis of enteritis and / or hepatitis in a subject, said method using the amount of D-amino acid in the intestinal tract as an indicator.

Description

Technical Field

[0005] ,

[0001] The present invention relates to drugs and foods for treating and / or preventing enteritis and / or hepatitis. The present invention also relates to a method for assisting the diagnosis of enteritis and / or hepatitis in a subject.

Background Art

[0002] It was thought that only L-amino acids constituted the living body, but it has been revealed that D-amino acids exist in trace amounts, and their various physiological functions and relationships with diseases have been reported. For example, it has been reported that D-Ser binds to the N-methyl-D-Aspartate (NMDA) receptor in the central nervous system in the brain and exhibits a physiological effect (Non-Patent Document 1).

[0003] It has also been reported that intestinal bacteria synthesize D-amino acids. For example, it has been reported that D-Ser synthesized by intestinal bacteria reaches the kidneys via the blood and reduces kidney damage (Non-Patent Document 2). It has also been reported that D-amino acids synthesized by intestinal bacteria are decomposed by D-amino acid oxidase (DAO) secreted from the host's intestinal tract, and the hydrogen peroxide generated in the process suppresses the growth of intestinal pathogenic bacteria (Non-Patent Document 3).

[0004] Further, Patent Document 1 reports that an extract of a Lactobacillus bacterium strain has an anti-inflammatory effect, and that the extract of the Lactobacillus bacterium strain contains D-amino acids (D-alanine, D-serine, D-glutamic acid, D-aspartic acid, etc.) (Patent Document 1). However, in none of the documents has it been reported that D-amino acids themselves can directly become drugs or foods capable of treating and / or preventing enteritis and / or hepatitis, and there is at least no finding that D-amino acids can be indicated as an active ingredient of drugs or foods alone. <​​​​​​​ [Patent Document 1] International Publication No. 2010 / 027344 [Non-patent literature]

[0006] [Non-Patent Document 1] FEBS Journal 275 (2008) 3514-3526. [Non-Patent Document 2] JCI Insight. 2018;3(20):e97957. [Non-Patent Document 3] Nature Microbiology vol. 1(2016), Article number: 16125. [Overview of the project] [Problems that the invention aims to solve]

[0007] The objective is to provide novel drugs and foods for the treatment and / or prevention of enteritis and / or hepatitis. It also aims to provide methods to aid in the diagnosis of enteritis and / or hepatitis. [Means for solving the problem]

[0008] 5-aminosalicylic acid preparations are believed to improve various symptoms of inflammatory bowel disease through their effects such as scavenging reactive oxygen species and inhibiting the production of leukotrienes.

[0009] As a result of diligent research, the inventors of this application have newly discovered that D-amino acids, which generate hydrogen peroxide in the intestinal tract in contrast to 5-aminosalicylic acid preparations, have an effect in suppressing enteritis and / or hepatitis.

[0010] Furthermore, as a result of diligent research, the inventors of this application have newly discovered that the amount of D-amino acids in the intestinal tract can serve as an indicator for diagnosing enteritis and / or hepatitis.

[0011] The present invention includes the following [1] to "7H": [1] A drug for treating and / or preventing enteritis and / or hepatitis, comprising a D-amino acid, a modified or derivative thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. [2] The agent according to [1], wherein the D-amino acid is at least one selected from the group consisting of D-Ala, D-Pro, D-Gln, D-Val, D-Leu, D-Phe, D-Lys, D-Trp, D-Glu, D-Ser and D-Asp. [3] The drug according to [1] or [2], wherein the enteritis is inflammatory bowel disease. [4] The drug described in [3], wherein the inflammatory bowel disease is ulcerative colitis. [5] The drug described in any one of the items [1] to [4], wherein the hepatitis is cholangitis. [6] The drug according to [5], wherein the cholangitis is primary sclerosing cholangitis (PSC).

[0012] [1A] A food composition for suppressing enteritis and / or hepatitis, containing a D-amino acid, a modified or derivative thereof, or a food-acceptable salt thereof as an active ingredient. [2A] The food composition for suppressing enteritis and / or hepatitis according to [2A], wherein the D-amino acid comprises at least one selected from the group consisting of D-Ala, D-Pro, D-Gln, D-Val, D-Leu, D-Phe, D-Lys, D-Trp, D-Glu, D-Ser, and D-Asp. [3A] The food composition according to [1A] or [2A], wherein the enteritis is inflammatory bowel disease. [4A] The food composition according to [3A], wherein the inflammatory bowel disease is ulcerative colitis. [5A] The food composition according to any one of [1A] to [4A], wherein the hepatitis is cholangitis. [6A] The food composition according to [5A], wherein the cholangitis is primary sclerosing cholangitis (PSC).

[0013] A food containing a food composition described in any one of the items [7A], [1A], to [6A].

[0014] [1B] D-amino acids, their modified or derivatives, or pharmaceutically acceptable salts thereof, for use in the treatment and / or prevention of enteritis and / or hepatitis. [2B] The D-amino acid according to [1B], a modified or derivative thereof, or a pharmaceutically acceptable salt thereof, comprising at least one selected from the group consisting of D-Ala, D-Pro, D-Gln, D-Val, D-Leu, D-Phe, D-Lys, D-Trp, D-Glu, D-Ser, and D-Asp. [3B] The D-amino acid, a modified or derivative thereof, or a pharmaceutically acceptable salt thereof, as described in [1B] or [2B], wherein the enteritis is an inflammatory bowel disease. [4B] The inflammatory bowel disease is ulcerative colitis, the D-amino acid described in [3B], its modified or derivative, or a pharmaceutically acceptable salt thereof. [5B] The hepatitis is cholangitis, and the D-amino acid, its modified or derivative, or a pharmaceutically acceptable salt thereof as described in any one of [1A] to [4A]. [6B] The D-amino acid described in [5B], its modified or derivative, or a pharmaceutically acceptable salt thereof, wherein the cholangitis is primary sclerosing cholangitis (PSC).

[0015] [1C] A method for treating and / or preventing enteritis and / or hepatitis in a subject, comprising administering to the subject an effective amount of a D-amino acid, a modified or derivative thereof, or a pharmaceutically acceptable salt thereof, for suppressing the onset of symptoms of enteritis and / or hepatitis in the subject or alleviating the symptoms of such symptoms. [2C] The method according to [1C], wherein the D-amino acid comprises at least one selected from the group consisting of D-Ala, D-Pro, D-Gln, D-Val, D-Leu, D-Phe, D-Lys, D-Trp, D-Glu, D-Ser, and D-Asp. [3C] The method according to [1C] or [2C], wherein the enteritis is an inflammatory bowel disease. [4C]The method according to [3C], wherein the inflammatory bowel disease is ulcerative colitis. [5C]The method according to any one of [1C] to [4C], wherein the symptoms of the enteritis are diarrhea, bloody stools and / or weight loss. [6C]The method according to any one of [1C] to [5C], wherein the hepatitis is cholangitis. [7C]The method according to [6C], wherein the cholangitis is primary sclerosing cholangitis (PSC).

[0016] [!D]Use of a D-amino acid, a modified form or derivative thereof, or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating and / or preventing enteritis and / or hepatitis. [2D]The use according to [1D], wherein the D-amino acid comprises at least one selected from the group consisting of D-Ala, D-Pro, D-Gln, D-Val, D-Leu, D-Phe, D-Lys, D-Trp, D-Glu, D-Ser and D-Asp. [3D]The use according to [1D] or [2D], wherein the enteritis is an inflammatory bowel disease. [4D]The use according to [3D], wherein the inflammatory bowel disease is ulcerative colitis. [[ID=!7]] [5D]The use according to any one of [1D] to [4D], wherein the hepatitis is cholangitis. [6D]The use according to [6D], wherein the cholangitis is primary sclerosing cholangitis (PSC).

[0017] [!E]A medicament or a food composition for suppressing hepatitis for treating and / or preventing hepatitis, which contains a D-amino acid, a modified form or derivative thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. <![CDATA[ ]]>[2E]The medicament or the food composition for suppressing hepatitis according to [1E], wherein the D-amino acid is at least one selected from the group consisting of D-Ala, D-Pro, D-Gln, D-Val, D-Leu, D-Phe, D-Lys, D-Trp, D-Glu, D-Ser and D-Asp. [3E] The drug or food composition for suppressing hepatitis according to [1E] or [2E], wherein the hepatitis is cholangitis. [4E] The drug or hepatitis-suppressing food composition according to [3E], wherein the cholangitis is primary sclerosing cholangitis (PSC).

[0018] [1F] A method for treating and / or preventing hepatitis in a subject, comprising administering to the subject an effective amount of a D-amino acid, a modified or derivative thereof, or a pharmaceutically acceptable salt thereof, for suppressing the onset of or alleviating the symptoms of hepatitis in the subject. [2F] The method according to [1C], wherein the D-amino acid comprises at least one selected from the group consisting of D-Ala, D-Pro, D-Gln, D-Val, D-Leu, D-Phe, D-Lys, D-Trp, D-Glu, D-Ser, and D-Asp. [3F] The method according to [1F] or [2F], wherein the hepatitis is cholangitis. [4F] The method according to [3F], wherein the cholangitis is primary sclerosing cholangitis (PSC).

[0019] [1G] Use of D-amino acids, their modifications or derivatives, or pharmaceutically acceptable salts thereof in the manufacture of drugs for the treatment and / or prevention of hepatitis. [2G] The use according to [1G], wherein the D-amino acid comprises at least one selected from the group consisting of D-Ala, D-Pro, D-Gln, D-Val, D-Leu, D-Phe, D-Lys, D-Trp, D-Glu, D-Ser, and D-Asp. [3G] The use according to [1G] or [2G], wherein the hepatitis is cholangitis. [4G] The use described in [3G], wherein the cholangitis is primary sclerosing cholangitis (PSC).

[0020] [1H] A method to aid in the diagnosis of enteritis and / or hepatitis, using the amount of D-amino acids in the intestinal tract as an indicator. [2H] The method according to [1H], using the amount of D-amino acids in the stool of the subject as an indicator. [3H] The method according to [1H] or [2H], wherein the D-amino acid comprises at least one selected from the group consisting of D-Ala, D-Pro, D-Gln, D-Val, D-Leu, D-Phe, D-Lys, D-Trp, D-Glu, D-Ser, and D-Asp. [4H] The method according to any one of [1H] to [3H], wherein the colitis is inflammatory bowel disease. [5H] The method according to [4H], wherein the inflammatory bowel disease is ulcerative colitis. [6H] The method according to any one of [1H] to [5H], wherein the hepatitis is cholangitis. [7H] The method according to [6H], wherein the cholangitis is primary sclerosing cholangitis (PSC). [Effects of the Invention]

[0021] The present invention makes it possible to provide novel drugs and foods for treating and / or preventing new types of enteritis and / or hepatitis, as well as methods for treating or preventing enteritis and / or hepatitis. Furthermore, the present invention makes it possible to provide methods for assisting in the diagnosis of target types of enteritis and / or hepatitis. [Brief explanation of the drawing]

[0022] [Figure 1] Figure 1 shows the effect of D-amino acids on inhibiting colitis in a DSS-induced colitis model mouse in one embodiment of the present invention. Figure 1A: Experimental schedule; Figure 1B: Change in body weight over time; Figure 1C: DAI (disease activity index) score on day 7 after the start of DSS administration; Figure 1D: Percentage of IL-17 positive cells in the colon on day 7 after the start of DSS administration. [Figure 2] Figure 2 shows the effect of D-amino acids on suppressing colitis in immunodeficient mice (RAG2KO) in one embodiment of the present invention. Figure 2A: Changes in body weight over time in immunodeficient mice (left panel) and body weight 8 days after the start of DSS administration (right panel); Figure 2B: DAI score 8 days after the start of DSS administration. [Figure 3]Figure 3 shows the inhibitory effect of various D-amino acids on enteritis in one embodiment of the present invention. Figure 3A: Experimental schedule; Figure 3B: Body weight on the 8th day after the start of DSS administration; Figure 3C: DAI score on the 8th day after the start of DSS administration. [Figure 4] Figure 4 shows the ratio of D-amino acids to L-amino acids in the feces of healthy individuals and patients with inflammatory bowel disease (IBD). [Figure 5] Figure 5 is a diagram showing the anti-inflammatory and preventive effects of D-amino acid administration in a mouse model of liver disease complicated with ulcerative colitis.

Mode for Carrying Out the Invention

[0023] Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings, but the present invention is not necessarily limited thereto. The objects, features, advantages, and ideas of the present invention will be apparent to those skilled in the art from the description in this specification, and those skilled in the art can easily reproduce the present invention from the description in this specification. The embodiments and specific examples of the invention described below show preferred embodiments of the present invention and are shown for the purpose of illustration or explanation, and the present invention is not limited thereto. It will be apparent to those skilled in the art that various modifications and variations can be made within the spirit and scope of the present invention disclosed in this specification based on the description in this specification.

[0024] <D-amino acid> In this disclosure, D-amino acids are optical isomers of L-amino acids, including D-Ala (D-alanine), D-Arg (D-arginine), D-Asn (D-asparagine), D-Asp (D-aspartic acid), D-Cys (D-cysteine), D-Gln (D-glutamine), D-Glu (D-glutamic acid), D-His (D-histidine), D-Ile (D-isoleucine), and D-Leu (D- This includes D-amino acids (D-leucine), D-Lys (D-lysine), D-Met (D-methionine), D-Phe (D-phenylalanine), D-Pro (D-proline), D-Ser (D-serine), D-Thr (D-threonine), D-Trp (D-tryptophan), D-Tyr (D-tyrosine), D-Val (D-valine), their modifications or derivatives, and their pharmaceutically or food-acceptable salts. Modified or derivative D-amino acids include, for example, D-α-amino-n-butyric acid, D-homophenylalanine, D-allo-isoleucine, D-norleucine, D-norvaline, D-ornithine, etc., but derivatives may include D-phenylglycine, which is a derivative of glycine that does not have optical isomers.

[0025] In the present invention, the pharmaceutically or food-acidically acceptable salt of a D-amino acid may be in any form, such as an acidic salt, a basic salt, or an amphoteric salt. Any physiologically acceptable salt can be used, such as hydrochloride, sulfate, nitrate, sodium salt, potassium salt, calcium salt, or ammonium salt.

[0026] In this invention, modified or derivative D-amino acids refer to those that alter hydrophobic or electrostatic properties, or those that can increase the concentration of D-amino acids in the blood or tissues after administration. Any modified or derivative D-amino acid can be used as long as it can adjust the concentration of D-amino acids in the blood or tissues to an optimal level.

[0027] Examples of D-amino acid derivatives include compounds in which the carboxyl group, amino group, or hydroxyl group of a D-amino acid is protected or substituted. The carboxyl group can be esterified, amidated, etc. The amino group can be amidated. The hydroxyl group can be etherified or esterified. Examples of modified or derivative D-amino acids, such as modified or derivative D-alanine, include D-alanine methyl ester, D-alanine ethyl ester, and peptides containing D-alanine, such as dipeptides, tripeptides, oligopeptides, and polypeptides. Also, examples of modified or derivative D-amino acids, such as modified or derivative D-serine, include D-serine methyl ester, D-serine ethyl ester, and peptides containing D-serine, such as dipeptides, tripeptides, oligopeptides, and polypeptides.

[0028] When peptides are used, they may consist only of D-amino acids as active ingredients, or they may consist of D-amino acids as active ingredients plus other amino acids, such as alanine, glycine, valine, leucine, isoleucine, threonine, cysteine, methionine, aspartic acid, glutamic acid, asparagine, glutamine, lysine, arginine, phenylalanine, tyrosine, tryptophan, histidine, etc., and these may be in D-form or L-form.

[0029] <enteritis> In this disclosure, enteritis is a general term for diseases that exhibit inflammation of the intestinal mucosa. While not specifically limited to these diseases, examples include infectious enteritis (e.g., enteritis caused by Staphylococcus aureus, Vibrio parahaemolyticus, Salmonella, Vibrio cholerae, etc.), Crohn's disease, and inflammatory bowel diseases including ulcerative colitis.

[0030] Inflammatory bowel disease (IBD) is a condition that includes Crohn's disease and ulcerative colitis, characterized by chronic inflammation in various parts of the digestive tract, causing diarrhea and abdominal pain, and characterized by repeated relapses and remissions.

[0031] Ulcerative colitis (UC) is an inflammatory disease of the large intestine characterized by erosions and ulcers in the mucous membrane of the large intestine. Characteristic symptoms include diarrhea with or without bloody stools and abdominal pain. The lesions spread continuously from the rectum upwards, and can extend from the rectum to the entire colon. Ulcerative colitis is classified as follows based on the extent and course of the lesions.

[0032] 1) Classification by extent of lesions: pancolitis, left-sided colitis, proctitis 2) Classification of disease stages: active phase, remission phase 3) Classification by severity: mild, moderate, severe, fulminant 4) Classification based on clinical course: relapse-remitting type, chronic persistent type, acute severe type, first attack type

[0033] The UC-DAI (Disease Active Index), which combines bowel movement frequency, amount of blood in stool, mucosal findings, and a physician's overall assessment, is used as an indicator of the severity of ulcerative colitis or improvement with medication.

[0034] Ulcerative colitis is a designated intractable disease. Traditionally, in mild cases, symptoms (inflammation) are controlled by administering 5-aminosalicylic acid preparations, which improve various symptoms of inflammatory bowel disease by removing reactive oxygen species and suppressing leukotriene production. If these are ineffective, steroids are administered, leukocyte apheresis is performed, and if these still do not subside, immunomodulators or biological agents such as TFα are administered. If these treatments are also ineffective, colectomy may be performed, and the development of new treatments is hoped for. The cause of ulcerative colitis is thought to be due to abnormalities in autoimmune responses or dietary habits, but it remains unknown.

[0035] <Hepatitis> In this disclosure, hepatitis is a general term for diseases in which inflammation of liver cells is caused by some factor. Depending on the cause, hepatitis can include viral hepatitis, alcoholic liver disease, non-alcoholic steatohepatitis, drug-induced liver injury, autoimmune hepatitis, and various types of cholangitis, and there are conditions that present with symptoms acutely or chronically. <Cholangitis> In this disclosure, cholangitis is a general term for diseases exhibiting inflammation of the bile ducts. Specific diseases are not limited, but examples include acute cholangitis that occurs when bacterial infection is added to the presence of gallstones in the common bile duct, or when there is impacted gallstones or stricture of the bile duct; primary sclerosing cholangitis (PSC). In particular, primary sclerosing cholangitis (PSC) is a disease characterized by patchy inflammation, fibrosis, and stricture of the bile ducts, the cause of which remains unknown. 80% of patients with primary sclerosing cholangitis (PSC) also have inflammatory bowel disease, particularly ulcerative colitis. In one embodiment, the present invention may treat or prevent cholangitis, such as primary sclerosing cholangitis (PSC).

[0036] <Medications and methods of administration for treating and / or preventing enteritis and / or hepatitis> One aspect of the present invention is a drug (or pharmaceutical composition) for treating and / or preventing enteritis and / or hepatitis, comprising a D-amino acid, a modified or derivative thereof, or a pharmaceutically acceptable salt thereof. In addition to the D-amino acid, its modified or derivative thereof, or a pharmaceutically acceptable salt thereof, the drug of one aspect of the present invention may also contain a pharmaceutically acceptable carrier, diluent, or excipient. In addition to the D-amino acid, its modified or derivative thereof, or a pharmaceutically acceptable salt thereof, the drug of the present invention may also contain further anti-enteritis agents, anti-hepatitis agents, etc. The drug of the present invention can be formulated in a dosage form suitable for its route of administration. For oral administration, dosage forms such as tablets, capsules, liquids, powders, granules, and chewable preparations may be designed; for parenteral administration, dosage forms such as injections, powders, and intravenous solutions may be designed. Furthermore, these formulations may contain various auxiliary agents used in pharmaceuticals, namely carriers and other auxiliary agents, such as stabilizers, preservatives, analgesics, flavoring agents, taste enhancers, fragrances, emulsifiers, fillers, and pH adjusters, and can be incorporated within a range that does not impair the effects of the composition of the present invention. In order to improve or maintain the effect as a pharmaceutical agent, the optical purity of the D-amino acid is preferably 50% or higher, and more preferably 90% or higher.

[0037] Furthermore, one aspect of the present invention is a method for administering an agent for treating and / or preventing enteritis, comprising a D-amino acid, a modified or derivative thereof, or a pharmaceutically acceptable salt thereof. Treatment of enteritis and / or hepatitis includes achieving remission (i.e., a state in which symptoms subside). Symptoms of enteritis include diarrhea, bloody stools, and / or weight loss, and if hepatitis persists, liver fibrosis may progress, potentially leading to cirrhosis and hepatocellular carcinoma.

[0038] This drug may be provided in dosage forms suitable for local administration (on the skin, by inhalation, enema, eye drops, ear drops, nasal, vaginal, etc.), enteral administration, or parenteral administration (intravenous, intra-arterial, transdermal, intramuscular injection, etc.), but enteral administration is preferred. Enteral administration includes oral administration, tube administration, and enema administration. Tube administration includes administration via nasogastric tube, gastrostomy, or duodenal fistula. Enema administration includes administration using suppositories or enemas. In any case, the dosage form of the drug is not particularly limited and may be liquid or solid, and can be manufactured according to the common technical knowledge of those skilled in the art. The specific method of administration is also not particularly limited and can be suitably administered according to the common technical knowledge of those skilled in the art.

[0039] The target of administration is not particularly limited, but it is preferably a human or a non-human vertebrate, more preferably a mammal such as a human or mouse, and even more preferably a human.

[0040] The dosage of D-amino acids as a drug or as a food composition can be set at any dose from the viewpoint of exerting an effect in treating or preventing enteritis and / or hepatitis. However, in the case of D-serine, it is preferable to appropriately adjust the concentration in the intestinal tract and / or liver, based on the following reports.

[0041] D-serine has been reported to cause nephrotoxicity in rats when administered intraperitoneally (Non-patent literature (1): Alexander WK, Am J Physiol Renal Physiol 2007, 293, F382-F390; Non-patent literature (2): M. Maekawa et al., Chem. Res. Toxicol. 2005, 18, 1678-1682; Non-patent literature (3): M. Orozco-Ibarra et al., Toxicology 229 (2007) 123-135; Non-patent literature (4): RE Williams et al., Toxicology 207 (2005) 179-190; Non-patent literature (5): RE Williams, EA Lock, Toxicology 201 (2004) 231-238). These documents disclose that nephrotoxicity is induced by oxidative stimulation and the action of DAO when administered at doses exceeding 250 mg / kg, for example, 800 mg / kg (Non-Patent Document (2)), 400 mg / kg (Non-Patent Document (3)), and 250 mg / kg (Non-Patent Documents (4) and (5)). Specifically, it is thought that D-serine is metabolized by the action of D-amino acid oxidase (DAO) in the proximal tubules, and that the reactive oxygen species generated in this reaction cause cell damage, leading to necrosis.

[0042] Non-patent document (1) describes a study in rats that investigated the effects of intraperitoneal administration of D-serine at concentrations of 0.25, 0.76, 2.54, and 7.6 mmol / kg on the kidneys. Since the molecular weight of D-serine is 105, these concentrations are converted to 26 mg / kg, 80 mg / kg, 267 mg / kg, and 800 mg / kg. When these concentrations of D-serine were administered intraperitoneally, the 26 mg / kg and 80 mg / kg concentrations showed no effect on amino acid excretion after 2 hours, indicating no toxicity. However, the 267 mg / kg and 800 mg / kg concentrations significantly increased amino acid excretion, demonstrating the renal toxicity of D-serine. Furthermore, when amino acid excretion after 4 hours was examined, no toxicity was observed at 26 mg / kg, but a considerably weak toxicity was observed at 80 mg / kg. Therefore, the no-observed-adverse-effect level (NOAEL) in rats can be set at 25, 30, 40, or 50 mg / kg. Therefore, the D-serine dose may be set to the amount of NOAEL, or, taking into account species differences, a safety factor of 10 may be set, and the upper limit of the dose may be set to 2.5 to 5.0 mg / kg / day. The safety factor can be set arbitrarily. Preferably, the upper limit of the dose is 5.0 mg / kg / day, more preferably 4.0 mg / kg / day, even more preferably 3.0 mg / kg / day, and particularly preferably 2.5 mg / kg.

[0043] In this example, by allowing mice to freely drink water containing 1% D-serine, a blood D-serine concentration of 100 nmol / mL was achieved, and at this concentration, the therapeutic effect of D-serine on enteritis and hepatitis was demonstrated. Although consideration of differences in species is necessary, not limited to theory, the effects of the present invention, i.e., the therapeutic effect on enteritis and hepatitis, can be exerted if the blood concentration of D-serine is 1 nmol / mL to 1 μmol / mL. The blood concentration after D-serine administration is preferably 5 nmol / mL or higher, more preferably 10 nmol / mL or higher, and even more preferably 50 nmol / mL or higher. On the other hand, the blood concentration of D-serine is preferably 1 μmol / mL or lower, more preferably 0.5 μmol / mL or lower, and even more preferably 0.1 μmol / mL or lower.

[0044] <Foods for suppressing enteritis and / or hepatitis> One aspect of the present invention relates to a food for suppressing enteritis and / or hepatitis, and a food containing such food, containing D-amino acids, their modified or derivatives, or food-acceptable salts thereof. A food for suppressing enteritis and / or hepatitis refers to a food labeled for consumption by individuals suffering from enteritis and / or hepatitis or those at risk of developing enteritis and / or hepatitis. The form of the food is not particularly limited, but it may be added to a daily diet or taken as a supplement or dietary aid. In this disclosure, food also includes beverages. Such foods may be provided as functional foods or health foods. D-amino acids, their modified or derivatives, or food-acceptable salts thereof, and raw materials containing them can be incorporated into any food. To improve or maintain the effectiveness of the food, the optical purity of the D-amino acids is preferably 50% or higher, and more preferably 90% or higher.

[0045] By consuming this food for suppressing enteritis and / or hepatitis, the symptoms of enteritis and / or hepatitis can be improved. It can also prevent enteritis and / or hepatitis and can be used, for example, as a digestive aid.

[0046] <Methods to assist in the diagnosis of enteritis and / or hepatitis> One aspect of the present invention provides a method to assist in the diagnosis of a target enteritis and / or hepatitis, using the amount of D-amino acids in the intestinal tract as an indicator. Through diligent research, the inventors have found that there are variations in the amount of D-amino acids in the stool of healthy individuals and patients with enteritis, particularly those with inflammatory bowel disease (IBD). This has led to the discovery that measuring the amount of D-amino acids in the target intestinal tract can be used to determine and diagnose the pathology of a target enteritis, such as inflammatory bowel disease, particularly ulcerative colitis. The pathology of enteritis can be determined by measuring the amount of D-amino acids in the intestinal contents, such as stool. Furthermore, this indicator can be used not only for diagnosing the pathology but also for diagnosing disease progression and prognosis, and for evaluating the effectiveness of treatment and medication. In this case, values ​​corrected for, for example, L-amino acids may be used. The D-amino acids used for determination may be at least one selected from the group consisting of, for example, D-Ala, D-Pro, D-Gln, D-Val, D-Leu, D-Phe, D-Lys, D-Trp, D-Glu, D-Ser, and D-Asp, or a combination of multiple D-amino acids may be used. Alternatively, the amount of L-amino acids corresponding to the selected D-amino acids may also be measured and the determination may be made using the corrected value, or the determination may be made by the ratio of the amount of D-amino acids to the amount of L-amino acids. Furthermore, the present invention may be used to determine and diagnose the pathology of enteritis, such as inflammatory bowel disease, particularly ulcerative colitis, and hepatitis, such as cholangitis, particularly primary sclerosing cholangitis (PSC), which occurs concurrently with it.

[0047] Subjects diagnosed with enteritis and / or hepatitis by the method of the present invention can be treated by administering or ingesting agents or food compositions for treating and / or preventing enteritis and / or hepatitis, as already described in other embodiments.

[0048] Furthermore, subjects whose enteritis and / or hepatitis status is determined by the method of the present invention may be treated with known therapies used for the treatment of enteritis, such as aminosalicylate preparations (e.g., mesalazine (5-ASA)), anti-TNF-α antibody preparations (e.g., infliximab), anti-IL-12 / 23p40 antibody preparations (e.g., ustekinumab), anti-α4β7 integrin antibody preparations (e.g., betrizumab), JAK inhibitors (e.g., tofacitinib citrate), steroids, immunomodulators (e.g., thiopurine preparations: azathioprine), immunosuppressants (e.g., calcineurin inhibitors: cyclosporine), glycyrrhizin preparations, synthetic disaccharides, antibacterial agents, antihypertensive agents, diuretics, albumin preparations, and carnitine. Treatment may be provided by the administration of pharmaceutical preparations, opioid potassium receptor agonists, carnitine, acetylcysteine, resins, dyslipidemia treatments (e.g., statins, small intestinal cholesterol transporter inhibitors, fibrates), thiazoline derivatives, oily contrast agents, choleretic agents (e.g., ursodeoxycholic acid), protease inhibitors, antithrombin, alcoholism treatments (e.g., cyanamide), HA vaccines, anti-HB human immunoglobulins, HB vaccines, anti-hepatitis virus drugs (e.g., interferon preparations, RNA polymerase inhibitors, protease inhibitors), nutritional therapy (e.g., intake of elemental nutritional formulas, digested nutritional formulas, semi-digested nutritional formulas, etc.), apheresis, or surgical intervention.

[0049] The present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples. [Examples]

[0050] Example 1: Inhibitory effect of D-amino acids on colitis in a DSS-induced colitis model.

[0051] 1. Method Mice (B6CL57) were given free access to either drinking water containing five D-amino acids (1% D-serine, 0.75% D-glutamic acid, 1% D-asparagine, 1% D-tryptophan, and 1% D-alanine) (shown as D-Amino in the figure) or drinking water containing five L-amino acids (1% L-serine, 0.75% L-glutamic acid, 1% L-asparagine, 1% L-tryptophan, and 1% L-alanine) (shown as L-Amino in the figure) for three weeks. Mice (control) were given free access to normal drinking water for three weeks. Chemical colitis was then induced in these mice by giving them free access to a 2% DSS (Dextran sulfate sodium) aqueous solution for seven days, and the severity of the colitis (body weight, pathology score (DAI score)) was measured (Figures 1B and C).

[0052] The DAI score was calculated by scoring each mouse based on three items: diarrhea symptoms (normal: 0, loose stools: 1, mild diarrhea: 2, moderate diarrhea: 3, severe diarrhea: 4); bloody stool symptoms (normal: 0, mild bloody stools: 1, moderate bloody stools: 2, severe bloody stools: 3, severe bloody stools with anal bleeding: 4); and percentage of weight loss (none: 0, 1-5%: 1, 5-10%: 2, 10-20%: 3, ≥20%: 4), and then summing the scores (minimum 0 points, maximum 12 points). Furthermore, the proportion of Th17 cells, which are IL-17-positive T cells that induce inflammation, was measured in the large intestine (Figure 1D).

[0053] 2.Results In the group of mice that consumed drinking water containing D-amino acids, weight loss was significantly suppressed from day 6 onwards, as shown in Figure 1B, and the DAI score on day 7 was significantly lower, as shown in Figure 1C, compared to the groups of mice that consumed drinking water containing L-amino acids or the groups that consumed regular drinking water.

[0054] Furthermore, as shown in Figure 1D, the proportion of IL-17-positive Th17 cells in the colon on day 7 was significantly lower in the mouse group that consumed drinking water containing D-amino acids compared to the mouse group that consumed drinking water containing L-amino acids. Th17 cells, which produce IL-17 and are present in the intestinal tract, are a cell population that increases in inflammatory bowel disease and other conditions, and are inflammation-inducing cells. Therefore, Th17 cells increase in the intestinal tract where inflammation is occurring, and decrease in the intestinal tract where inflammation is suppressed. Consequently, this indicates that inflammation caused by DSS intake was suppressed in the mouse group that consumed drinking water containing D-amino acids.

[0055] Thus, in the group of mice that consumed drinking water containing D-amino acids, the exacerbation of enteritis was significantly suppressed.

[0056] Example 2: Inhibitory effect of D-amino acids on enteritis in immunodeficient mice

[0057] 1. Method Immunodeficient mice (RAG2-deficient mice; RAG2KO) were given free access to drinking water containing five D-amino acids (1% D-serine, 0.75% D-glutamic acid, 1% D-asparagine, 1% D-tryptophan, and 1% D-alanine) or five L-amino acids (1% L-serine, 0.75% L-glutamic acid, 1% L-asparagine, 1% L-tryptophan, and 1% L-alanine) for three weeks. Immunodeficient mice were also given free access to normal drinking water for three weeks. Chemical colitis was induced in these mice by providing them with free access to a 2% DSS aqueous solution for seven days, and the severity of the colitis (body weight, pathology score (DAI score)) was measured (Figures 2A and B).

[0058] 2.Results In RAG2KO mice, the group of mice that consumed drinking water containing D-amino acids showed significantly suppressed exacerbation of colitis compared to the groups of mice that consumed drinking water containing L-amino acids or regular drinking water, similar to the results obtained in Example 1. This indicates that D-amino acids have a colitis-suppressing effect even in mice that lack acquired immune function.

[0059] Example 3: Inhibitory effect of a single D-amino acid on colitis in a DSS-induced colitis model mouse.

[0060] 1. Method Mice (B6CL57) were given free access to drinking water containing five types of D-amino acids (1% D-serine, 0.75% D-glutamic acid, 1% D-asparagine, 1% D-tryptophan, and 1% D-alanine) or a single D-amino acid (1% D-serine, 0.75% D-glutamic acid, 1% D-asparagine, 1% D-tryptophan, or 1% alanine) for three weeks. Chemical colitis was then induced by free access to a 2% DSS aqueous solution for seven days, and the severity of the colitis (body weight, pathology score (DAI score)) was measured (Figure 3A).

[0061] 2.Results In terms of both weight loss suppression effect and DAI score, drinking water containing D-Ser, D-Ala, and D-Asp D-amino acids had a comparable enteritis-suppressing effect to drinking water containing five D-amino acids, while drinking water containing D-Trp and D-Glu D-amino acids suppressed enteritis significantly more effectively than drinking water containing five D-amino acids (Figures 3B and 3C).

[0062] Example 4: Pathological indicators of inflammatory bowel disease (IBD) using D-amino acids in the stool of patients.

[0063] Stool samples from healthy individuals and inflammatory bowel disease (IBD) patients were homogenized with Milli-Q water, methanol was added, and amino acids were extracted by centrifugation. These amino acids were modified with NBD-F, and the data were separated and quantified by liquid chromatography for analysis. The D-amino acids in the stool of IBD patients (corrected using L-amino acids) were characteristically lower than those in healthy individuals, suggesting that the amount of D-amino acids in stool is useful for understanding the disease state, determining drug dosages, and evaluating the effectiveness of treatment and medications.

[0064] Example 5: Inflammatory effect in a DDC-induced liver disease model

[0065] 1. Method In germ-free mice (GFs) that were given free access to drinking water containing five D-amino acids (1% D-serine, 0.75% D-glutamic acid, 1% D-asparagine, 1% D-tryptophan, and 1% D-alanine) (D-mix in the figure), feces from patients with ulcerative colitis (UC) and primary sclerosing cholangitis (PSC) were transplanted. In these model mice, cholangitis was induced with 3,5-diethoxvcarbonyl-1,4-dihydrocollidine (DDC), and in similar model mice given free access to normal drinking water (Figure 5A), total bilirubin (T-Bill) and aminotransferase (ALT), liver disease markers, were measured in the blood.

[0066] 2.Results In the group of mice that consumed drinking water containing D-amino acids, liver disease markers were significantly lower, and hepatitis was suppressed and prevented compared to the group of mice that consumed regular drinking water (Figure 5B, C). [Industrial applicability]

[0067] The present invention makes it possible to provide novel drugs and foods for treating and / or preventing enteritis and / or hepatitis, as well as methods for treating or preventing enteritis and / or hepatitis.

Claims

1. A drug used to treat and / or prevent enteritis and / or hepatitis when enteritis and / or hepatitis is diagnosed, based on the characteristically low amount of D-amino acids or their ratio to L-amino acids in the subject's body compared to a healthy control group, It contains D-amino acids or pharmaceutically acceptable salts thereof as active ingredients, The aforementioned enteritis is inflammatory bowel disease, The aforementioned hepatitis is cholangitis. A drug in which the D-amino acid is selected from the group consisting of D-serine, D-glutamic acid, D-aspartic acid, D-tryptophan, D-alanine, D-leucine, and D-lysine.

2. The agent according to claim 1, wherein the inflammatory bowel disease is ulcerative colitis.

3. The agent according to claim 1 or 2, wherein the cholangitis is primary sclerosing cholangitis (PSC).

4. The drug according to any one of claims 1 to 3, wherein the patient determines enteritis and / or hepatitis using the ratio of the amount of D-amino acids to the amount of L-amino acids in the body of the subject, or a corrected value obtained by correcting the amount of D-amino acids by the amount of L-amino acids.

5. A drug according to any one of claims 1 to 4, wherein the amount of D-amino acids in the subject's body decreases, and the subject is determined to have enteritis and / or hepatitis.

6. Enteritis and / or hepatitis are determined based on the fact that the amount of D-amino acids in the subject's body or the ratio of D-amino acids to L-amino acids is characteristically lower than that of a healthy control group, and if enteritis and / or hepatitis is determined, the use of D-amino acids or pharmaceutically acceptable salts thereof in the manufacture of drugs used to treat and / or prevent enteritis and / or hepatitis, The aforementioned enteritis is inflammatory bowel disease, The aforementioned hepatitis is cholangitis. The D-amino acid is selected from the group consisting of D-serine, D-glutamic acid, D-aspartic acid, D-tryptophan, D-alanine, D-leucine, and D-lysine.

7. The use according to claim 6, wherein the inflammatory bowel disease is ulcerative colitis.

8. The use according to claim 6 or 7, wherein the cholangitis is primary sclerosing cholangitis (PSC).

9. The use according to any one of claims 6 to 8, wherein the ratio of the amount of D-amino acids to the amount of L-amino acids in the subject's body, or a corrected value obtained by correcting the amount of D-amino acids by the amount of L-amino acids, is used to determine whether enteritis and / or hepatitis have occurred.

10. The use according to any one of claims 6 to 9, wherein when the amount of D-amino acids in the subject's body decreases, it is determined that the subject has enteritis and / or hepatitis.

11. A food composition for use in improving enteritis and / or hepatitis, in which enteritis and / or hepatitis is determined based on the characteristically low amount of D-amino acids or their ratio to L-amino acids in the subject's body compared to a healthy control group, and which contains D-amino acids or food-acceptable salts thereof as active ingredients, The aforementioned enteritis is inflammatory bowel disease, The aforementioned hepatitis is cholangitis. A food composition in which the D-amino acid is selected from the group consisting of D-serine, D-glutamic acid, D-aspartic acid, D-tryptophan, D-alanine, D-leucine, and D-lysine.

12. The food composition according to claim 11, wherein the inflammatory bowel disease is ulcerative colitis.

13. The food composition according to claim 11 or 12, wherein the cholangitis is primary sclerosing cholangitis (PSC).

14. The food composition according to any one of claims 11 to 13, wherein the determination of enteritis and / or hepatitis is made using the ratio of the amount of D-amino acids to the amount of L-amino acids in the subject's body, or a corrected value obtained by correcting the amount of D-amino acids by the amount of L-amino acids.

15. A food composition according to any one of claims 11 to 14, wherein if the amount of D-amino acids in the subject's body decreases, it is determined that the subject has enteritis and / or hepatitis.

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