Capsaicin in the treatment of leaky gut (intestinal wall leakage)

Abstract

The present invention relates to the treatment of leaky gut. More specifically, the present invention provides compounds and compositions thereof for use in methods for treating leaky gut and methods for treating asymptomatic increases in intestinal permeability. The present invention further provides compounds and compositions thereof for use in methods for treating leaky gut syndrome. The present invention further provides compounds and compositions thereof for use in methods for ameliorating at least one symptom associated with leaky gut syndrome. The present invention also relates to methods for treating leaky gut, methods for treating leaky gut syndrome, and methods for ameliorating at least one symptom associated with leaky gut syndrome.

Description

Technical Field

[0001] The present invention relates to the treatment of leaky gut (intestinal wall leakage). More particularly, the present invention provides compounds and compositions thereof for use in methods of treating leaky gut and methods of treating asymptomatic increases in intestinal permeability. The present invention further provides compounds and compositions thereof for use in methods of treating leaky gut syndrome. The present invention further provides compounds and compositions thereof for use in methods of improving at least one symptom associated with leaky gut syndrome. The present invention also relates to methods of treating leaky gut, methods of treating leaky gut syndrome, and methods of improving at least one symptom associated with leaky gut syndrome.

Background Art

[0002] The gastrointestinal tract, the tube from mouth to anus present in all vertebrates and most invertebrates, contains all the organs of the digestive system. It is the most extensive barrier separating the internal environment of the body from the external environment; the lumen of the gastrointestinal tract is outside the animal's body, and the cells lining the tube create a barrier between the body and the outside world. Thus, the main role of the gastrointestinal tract is nutrient extraction, but it also forms an important part of the immune system and prevents pathogens such as viruses, molds, and bacteria from entering the blood and lymphatic circulatory systems.

[0003] It is well known that in critically ill patients, loss of intestinal integrity leads to the translocation of bacteria into the circulation and can ultimately lead to multiple organ failure and death. More recently, increasing attention has been focused on the importance of intestinal barrier integrity in non-critically ill subjects. It has been suggested that toxins can cross the epithelial barrier from the intestinal lumen to the host, leading to local and systemic immune responses. Various pathways have been discussed, including increased paracellular permeability of the intestinal mucosa. Such asymptomatic increases in intestinal permeability are often referred to as leaky gut, and the various health problems thought to be caused by this phenomenon are collectively referred to as leaky gut syndrome.

[0004] The gut is a habitat for a broad range of bacteria called the gut microbiota, containing approximately 4,000 different bacterial strains that play diverse roles in maintaining immune health and metabolism. Leaky gut syndrome may involve an imbalance in the gut microbiota, and one theory suggests that this imbalance triggers an immune response in the body, which in turn leads to increased inflammation and intestinal permeability in the gut. Frequently suggested risk factors for leaky gut syndrome, including physiological stressors such as anxiety, the use of nonsteroidal anti-inflammatory drugs (NSALDs), alcohol consumption, and dietary components including emulsifiers and other food additives, lend credibility to the theory of the microbiota's involvement in leaky gut, as it is known that these factors also affect the balance of the gut microbiota.

[0005] Leaky gut is thought to lead to increased systemic entry of pathogenic bacteria and bacterial toxins into the systemic circulation, with recurrent episodes triggering systemic inflammation and causing numerous diseases. Even certain bacteria normally present in a healthy digestive tract are thought to become pathogenic during leaky gut. Quorum sensing among gut bacteria may be a major factor in leaky gut and is suggested to influence its onset and development.

[0006] Increased intestinal permeability is widely accepted to be associated with certain gastrointestinal conditions such as celiac disease, Crohn's disease, irritable bowel syndrome (IBS), and inflammatory bowel disease (IBD). However, it is unclear whether leaky gut is a cause or symptom of any of these conditions, or whether the observed association can be explained by leaky gut syndrome, which frequently occurs concurrently with the aforementioned conditions or diseases. Furthermore, it has been suggested that leaky gut or leaky gut syndrome may also be associated with a wide range of other diseases, including autoimmune diseases (lupus, type 1 diabetes, multiple sclerosis, autoimmune hepatitis), chronic fatigue syndrome, fibromyalgia, arthritis, allergies, asthma, polycystic ovary syndrome, obesity, and even autism and mental disorders. Such causes and effects have not yet been confirmed in clinical trials in humans.

[0007] In addition to its potential contribution to other diseases, leaky gut syndrome is thought to cause symptoms including chronic diarrhea, constipation, bloating, gas, and stomach cramps; skin problems such as acne, rashes, and eczema; food sensitivity and nutritional deficiencies; throbbing and pain including joint pain; and headaches, confusion, difficulty concentrating, and fatigue. These symptoms may contribute to chronic inflammation throughout the body.

[0008] While various restrictive diets, nutritional supplements, and probiotics have been proposed, there is little evidence that the treatments offered for leaky gut are beneficial. Antibiotics have not been found to be useful; in fact, leaky gut appears to be exacerbated by antibiotics. None of the proposed treatments have been sufficiently tested to determine whether they are safe and effective for this purpose, and there is a lack of official recommendations regarding regimens for treating or managing the symptoms of leaky gut syndrome. Therefore, novel compounds and methods for treating leaky gut are needed. [Overview of the project]

[0009] The inventors have discovered that compounds of formula I can have a beneficial effect on leaky gut, and therefore these compounds are useful in methods for treating leaky gut and / or leaky gut syndrome, and in methods for improving at least one symptom associated with leaky gut syndrome. The inventors have found that these compounds inhibit quorum sensing, which is considered to be a major factor in leaky gut. It is strongly suggested that inhibiting bacterial quorum sensing can improve intestinal barrier function by preventing signaling that promotes individual bacteria to become pathogenic, for example, by forming tertiary structures such as biofilms, and thus positively influencing leaky gut. Compounds of formula I are promising candidates for the treatment of leaky gut, leaky gut syndrome, and symptoms associated with leaky gut syndrome.

[0010] In one embodiment, the present invention provides a compound of formula I or a pharmaceutically acceptable salt or solvate thereof for use in a method of treating leaky gut, [ka] During the ceremony, X is selected from oxygen and sulfur. R is selected from the group including cyclohexyl, phenyl, and substituted phenyl; The substituted phenyl is substituted at any one or more positions with one to five identical or different substituents selected from the group consisting of fluoro; chloro; bromo; iodo, cyano, nitro, trifluoromethyl, C1-C6 linear and branched alkoxy, C1-C6 sulfoxy, -S-C1-C6 alkyl, C1-C6 linear and branched alkyl, alkenyl and alkynyl, C2-C6 linear and branched alkenyl, C2-C6 linear and branched alkynyl, C1-C6 fluoroalkyl, chloroalkyl, bromoalkyl, and iodoalkyl, COO-C1-C6 alkyl, O(CO)-C1-C6 alkyl, NH-C1-C6 alkyl, N(C1-C6 alkyl)2, CON(C1-C6 alkyl)2, and NH(CO)-C1-C6 alkyl; R' is selected from the group including hydrogen, C1-C6 linear and branched alkyl groups, and C3-C6 cycloalkyl groups, and R'' is selected from the group including hydrogen, benzyl, and acetyl.

[0011] In another embodiment, the present invention provides a compound of formula I for use in a method of treating asymptomatic increase in intestinal permeability in subjects experiencing one or more symptoms associated with leaky gut syndrome and showing a significant deviation in the level of at least one biomarker compared to normal levels of at least one biomarker associated with leaky gut, leaky gut syndrome and / or increased intestinal permeability.

[0012] In another embodiment, the present invention provides a composition comprising a compound of formula I or a pharmaceutically acceptable salt or solvate thereof for use in a method of treating leaky gut.

[0013] In another embodiment, the present invention provides a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome, comprising the step of administering to a subject an effective amount of a compound of formula I or a pharmaceutically acceptable salt or solvate thereof, or a composition comprising an effective amount of a compound of formula I or a pharmaceutically acceptable salt or solvate thereof.

[0014] In another aspect, the present invention provides the use of a compound of formula I or a pharmaceutically acceptable salt or solvate thereof, or a composition comprising an effective amount of a compound of formula I or a pharmaceutically acceptable salt or solvate thereof, for inhibiting quorum sensing, inhibiting quorum sensing in the gut, treating leaky gut, treating leaky gut syndrome, or improving at least one symptom associated with leaky gut syndrome.

[0015] In another embodiment, the present invention provides a method for treating leaky gut, leaky gut syndrome, or increased intestinal permeability, comprising the step of administering an effective amount of a compound of formula I or a pharmaceutically acceptable salt or solvate thereof to a subject.

[0016] In another embodiment, the present invention provides the use of a compound of formula I or a pharmaceutically acceptable salt or solvate thereof for inhibiting quorum sensing. [Brief explanation of the drawing]

[0017] [Figure 1] Figure 1 shows the time-bacterial curves of different concentrations of phenylcapsaicin against Salmonella ATCC14028. [Figure 2]Figure 2 shows the time-kill curves of different concentrations of phenylcapsaicin against Listeria monocytogenes ATCC11915. [Figure 3] Figure 3 shows the time-kill curves of different concentrations of phenylcapsaicin against Campylobacter jejuni ATCC11168.

Mode for Carrying Out the Invention

[0018] Unless otherwise defined, all technical terms, notations and other scientific or technical terms used herein are intended to have the meanings commonly understood by those skilled in the art to which the present invention pertains. In some cases, terms with commonly understood meanings are defined herein for the purpose of clarification and / or ease of reference, and including such definitions herein should not necessarily be construed as representing a substantial difference from what is commonly understood in the art.

[0019] The term "capsaicin" as used herein refers to a hepta-6-ene derivative of capsaicin ((E)-N-(4-hydroxy-3-methoxybenzyl)-8-methylnon-6-enamide) of general formula a, wherein Y represents any substituent and Z represents any one or more substituents at any one or more positions on the phenyl ring.

Chemical formula

[0020] The reader is aware that the nomenclature used for capsaicin derivatives in the art is not consistent and that the term "capsaicin" is used, but for ease of reading, when referring to a group of common compounds, the well-established common name "phenylcapsaicin" is used to refer to N-[(4-hydroxy-3-methoxyphenyl)methyl]-7-phenylhepta-6-ynamide (Structure V), and "phenylcapsaicins" should be noted to be used to refer to its derivatives.

[0021] As used herein, the term "thioamide capsaicin" refers to a hepta-6-yne derivative of capsaicin containing a thioamide group rather than an amide group of general formula b, wherein Y represents any substituent and Z represents any one or more substituents at any one or more positions on the phenyl ring.

Chemical formula

[0022] As will be appreciated, any of the compounds described herein may be provided in the form of its pharmaceutically acceptable salts or solvates. Procedures for salt formation and solvate formation are conventional in the art.

[0023] As used herein, the terms linear and branched alkyl, alkenyl, alkynyl, and alkoxy include all such substituents of a given chain length, whether cyclic or including substituents containing a ring.

[0024] As used herein, the term “derivative” refers to a molecule that differs in chemical structure from its parent compound. Examples of derivatives include, but are not limited to, homologs that progressively differ from the parent's chemical structure, such as differences in the length of aliphatic chains; molecular fragments; structures that differ from the parent compound in one or more functional groups, for example, by transforming one or more functional groups of the parent compound; changes in the ionization state of the parent, such as ionization of an acid to a conjugate base; isomers, including positional isomers, geometric isomers, and stereoisomers; and combinations thereof.

[0025] The terms “to treat,” “treatment,” and “therapy” (and their grammatical variations) are used interchangeably herein and refer to: 1) inhibiting a disease, e.g., prevention of the disease (i.e., preventive measures, prevention of further progression of the pathology and / or overall symptoms), or 2) alleviating the symptoms of a disease; or 3) improving a disease, e.g., improving a disease, condition, or disorder (i.e., reversing the pathology and / or overall symptoms) in an object experiencing or exhibiting the pathology or overall symptoms of the disease; or 3) improving a disease, e.g., improvement of the disease, condition, or disorder (i.e., reversing the pathology and / or overall symptoms). These terms may be used in relation to the use and / or administration of pharmaceuticals, active pharmaceutical ingredients (APIs), food additives, supplements, health supplements, dietary supplements, over-the-counter (OTC) supplements, medical foods, and / or pharmaceutical-grade supplements.

[0026] As used herein, the term “composition” means a mixture of one or more compounds according to the present invention and one or more further chemical components in any formulation.

[0027] As used herein, the terms “administer,” “dosage,” and “administer” mean (1) providing, giving, administering and / or prescribing a preparation, preparation or composition according to the Disclosure by a healthcare professional or an authorized agent thereof, or under their direction, or by self-administration, and (2) introducing a preparation, preparation or composition according to the Disclosure into a subject, ingesting or consuming it by a subject.

[0028] As used herein, “Subject” means any human or non-human animal selected for treatment or therapy, and may include and be limited to “Patient.” Neither of these terms should be construed as requiring constant or otherwise supervision by a medical professional (e.g., physician, nurse, nurse practitioner, physician's assistant, hospital janitor, clinical research coordinator, etc.) or a scientific researcher.

[0029] As used herein, the term “therapeutic dose” means the amount of the compound according to the present invention that is effective in producing the desired therapeutic effect in a subject with a reasonable benefit / risk ratio applicable to any treatment. The therapeutic dose may vary depending on the route of administration and the form of drug delivery.

[0030] As used herein, the term “pharmaceutically acceptable” means that the compound must be physiologically acceptable to the recipient and, if it is part of a composition, must be compatible with the other components of the composition.

[0031] As used herein, the term "prodrug" means any compound that is converted to any of the compounds according to the present invention under physiological conditions.

[0032] As used herein, the term “symptom” means a disease, any physical disability, or any pathological phenomenon or deviation from normal of structure, function or sensation observed in or by an object, whether subjective or objective. As used herein, the term “improvement of symptoms” or any variation thereof means the alleviation or improvement of such symptoms.

[0033] As used herein, the term “leaky gut” refers to an asymptomatic increase in intestinal permeability. The symptoms caused by leaky gut are called “leaky gut syndrome.”

[0034] Bacteria become pathogenic, and their numbers in the host increase significantly through quorum sensing (QS). Quorum sensing is an intercellular communication mechanism that leads to differential gene expression in response to high population density. This process causes certain bacterial species to form biofilms, where individual cells begin to act as one larger tertiary organism. This process is mediated by the cell density-dependent expression of hormone-like compounds called autoinducers. Several processes involved in the successful establishment of bacterial infection are mediated by quorum sensing and the expression of pathogenic genes. Therefore, inhibiting quorum sensing is considered a suitable target for reducing the population of pathogenic bacteria in a host without antimicrobial activity.

[0035] The relationship between quorum sensing and biofilm formation is well established; see, for example, the 2005 review article “Sociomicrobiology: the connections between quorum sensing and biofilms” (Parsek and Greenberg, Trends Microbiol. 2005, 13(1), 27).

[0036] Based on the premise that leaky gut syndrome involves an imbalance in the gut microbiota, the theory that leaky gut can be treated by limiting quorum sensing has recently gained increasing support. Targeting bacterial quorum sensing has been shown to be promising in improving gut barrier function, lending credence to this theory. For example, in (Mol.Med.Rep.2019,19,4057), Adiliaghdam et al. demonstrated a robust relationship between inhibition of the important Pseudomonas aeruginosa quorum sensing system and the regulation of gut integrity after burns and burn wound infections. The results disclosed there provide evidence that silencing the Pseudomonas aeruginosa quorum sensing transcription factor MvfR leads to a less pronounced induction of gut barrier damage, and the results presented in the paper show that inhibition of this quorum sensing system reduces increased intestinal permeability.

[0037] Hepta-6-in derivatives of capsaicin, referred to herein as capsaicin(plural), are valuable compounds with a variety of potential applications. Distinguished from the natural compound capsaicin by the alkyne moiety that replaces the alkene moiety of capsaicin, these synthetic capsaicin derivatives have found use in a variety of fields, including the food industry, agriculture, pharmacology, and marine antifouling coatings. Phenylcapsaicin(V), perhaps the most widely used derivative, has been shown to have low systemic toxicity and be safe in terms of gene mutation and chromosomal damage (Rage Paulsen et al., Toxicology Research and Application 2018,2,1), and has been tested and deemed safe by the European Food Safety Authority (EFSA NDA Panel et al., EFSA Journal 2019,17(6),e05718).

[0038] Natural capsaicin is widely suspected of causing or contributing to leaky gut and leaky gut syndrome, and those affected are often advised to avoid foods containing capsaicin.

[0039] Surprisingly, the inventors have found that the compound of structure I is useful in a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome. [ka]

[0040] Our experiments demonstrated that the compound is a dose-dependent quorum sensing inhibitor at doses lower than the sublethal antimicrobial dose. Specifically, phenylcapsaicin(V) at doses of 61.5, 31.25, 15.625, and 7.81 μg / ml showed dose-responsive quorum sensing inhibitory activity ranging from strong to weak, but exhibited antimicrobial activity at higher doses (500 / 250 / 125 μg / ml).

[0041] The natural compound capsaicin did not exhibit quorum sensing inhibitory activity at non-antimicrobial doses (31.25 / 15.625 / 7.81 μg / ml), further reassuring the unexpected nature of the quorum sensing inhibitory activity of the compound of structure I for use in the method for treating leaky gut, the method for treating leaky gut syndrome, or the method for improving at least one symptom associated with leaky gut syndrome according to the present invention.

[0042] The results from these experiments indicate that the compound of structure I has a mechanism of action at sublethal antimicrobial doses that reduces bacterial pathogenicity and thus improves leaky gut syndrome. This effect is entirely unexpected; while the use of capsaicin(s) as TRPV1 agonists with a completely unrelated mechanism of action has been previously disclosed, the use of the compound of structure I to inhibit quorum sensing for leaky gut or in other therapeutic methods has not been demonstrated.

[0043] The fact that the compound inhibits quorum sensing at sublethal antimicrobial doses is particularly important. Lethal antimicrobial effects are undesirable and may even be harmful for compounds used in the treatment of leaky gut, as such effects would undoubtedly cause further imbalance in the gut microbiota, likely leading to a worsening of leaky gut. This is supported by evidence that antibiotics can exacerbate leaky gut syndrome.

[0044] Therefore, in the method for treating leaky gut, the method for treating leaky gut syndrome, or the method for improving at least one symptom associated with leaky gut syndrome according to the present invention, the dosage and administration regimen are always selected to avoid any lethal antimicrobial effect, taking into account the half-life of the compound according to the present invention as necessary. Lethal antimicrobial effects are avoided by selecting a sufficiently low dose of compound I, as is known to those skilled in the art, and / or based on the findings in the experiments presented in Example 2 or similar experiments. In this way, its beneficial effects, such as the gut microbiota and thus nutrient absorption, are not interfered with as much as possible.

[0045] The patient group of the present invention includes subjects suffering from leaky gut and / or leaky gut syndrome, for example, patients diagnosed with or suspected of having leaky gut or leaky gut syndrome. Another patient group of the present invention includes subjects experiencing at least one symptom associated with leaky gut syndrome. In some embodiments, subjects are selected from at least one of these patient groups.

[0046] The subject may be a human or a non-human animal, such as a human or non-human mammal, preferably a human patient. The subject may be male or female. In some embodiments of the present invention, the subject is an adult (i.e., 18 years of age or older). In certain embodiments, the subject is elderly. In certain embodiments, the subject is not elderly.

[0047] Leaky gut shares many of its symptoms with other health conditions, which can make it difficult to identify the condition. This invention relates to both the treatment of leaky gut or leaky gut syndrome itself in subjects, and the improvement of at least one symptom associated with leaky gut syndrome. In some embodiments, subjects are selected from a group of subjects who experience one or more symptoms associated with leaky gut syndrome, and the one or more symptoms are not attributable to other clinical diagnoses such as celiac disease, IBD, ulcerative colitis, or Crohn's disease.

[0048] In other embodiments, subjects are selected from a group of subjects experiencing one or more symptoms associated with leaky gut syndrome and further exhibiting a significant deviation in the level of at least one biomarker compared to normal levels of at least one biomarker associated with leaky gut, leaky gut syndrome, and / or increased intestinal permeability. In certain embodiments, the latter group is further limited to subjects who have not been diagnosed with celiac disease, IBD, ulcerative colitis, or Crohn's disease.

[0049] One such biomarker is zonulin. Studies and clinical trials of the protein zonulin and the zonulin signaling pathway have demonstrated the clinical efficacy of zonulin as a biomarker of intestinal permeability. Zonulin has been found to increase the permeability of the epithelial layer of the small intestine by reversibly regulating intercellular tight junctions. Dysregulation of the zonulin signaling pathway disrupts normal intestinal barrier function and alters the immune response. As a result, high levels of serum zonulin may indicate the presence of increased intestinal permeability. Other related biomarkers include enteric fatty acid-binding protein (I-FABP), soluble CD14, interleukin-6 (IL-6), and lipopolysaccharide (LPS).

[0050] Compounds for use in the treatment method for leaky gut according to the present invention can be obtained commercially or by any procedure known to those skilled in the art. Non-limiting examples of procedures for obtaining the compounds according to the present invention are disclosed by the applicant in European Patent No. 1670310 and Norwegian Patent Application No. 20200333.

[0051] In one embodiment, the present invention provides a compound of formula I or a pharmaceutically acceptable salt or solvate thereof for use in a method of treating leaky gut. [ka]

[0052] In a further embodiment, the present invention provides a compound of formula I or a pharmaceutically acceptable salt or solvate thereof for use in a method for treating leaky gut syndrome.

[0053] In a further embodiment, the present invention provides a compound of formula I or a pharmaceutically acceptable salt or solvate thereof for use in a method of improving at least one symptom associated with leaky gut syndrome in a subject, the at least one symptom not attributable to another clinical diagnosis.

[0054] In some embodiments, the subjects do not have celiac disease, IBD, ulcerative colitis, or Crohn's disease.

[0055] In a further embodiment, the present invention provides a compound of formula I or a pharmaceutically acceptable salt or solvate thereof for use in a method of improving at least one symptom associated with leaky gut syndrome in a subject, wherein the subject exhibits a significant deviation in the level of at least one biomarker compared to a normal level of at least one biomarker associated with leaky gut, leaky gut syndrome and / or increased intestinal permeability.

[0056] In some embodiments, at least one biomarker is selected from the list of zonulin, I-FABP, soluble CD14, IL-6, LPS, and any combination thereof. In certain embodiments, at least one biomarker is zonulin.

[0057] In some embodiments, the normal level is the average level in a population. In some embodiments, the normal level is the average level for a subject. In some embodiments, the normal level is determined by a physician based on the physician's common general knowledge.

[0058] In some embodiments, the subjects do not have celiac disease, IBD, ulcerative colitis, or Crohn's disease.

[0059] As discussed herein, leaky gut and leaky gut syndrome are associated with increased permeability, such as increased intestinal permeability. In another embodiment, the present invention provides compounds of formula I for use in a method of treating increased intestinal permeability in subjects experiencing one or more symptoms associated with leaky gut syndrome and showing a significant deviation in the level of at least one biomarker compared to normal levels of at least one biomarker associated with leaky gut, leaky gut syndrome and / or increased intestinal permeability.

[0060] Increased intestinal permeability may be a pathological increase in intestinal permeability. Preferably, the increase in intestinal permeability is an asymptomatic increase in intestinal permeability.

[0061] In some embodiments, the present invention provides compounds of formula I for use in a method of treating asymptomatic increase in intestinal permeability in subjects experiencing one or more symptoms associated with leaky gut syndrome and showing a significant deviation in the level of at least one biomarker compared to the normal level of at least one biomarker associated with increased intestinal permeability.

[0062] The biomarker may be, for example, any of the biomarkers disclosed above, preferably zonulin.

[0063] In some embodiments, the subjects are diagnosed with leaky gut and / or leaky gut syndrome. In some embodiments, the subjects are not diagnosed with celiac disease, IBD, ulcerative colitis, or Crohn's disease.

[0064] For both methods of improving at least one symptom associated with leaky gut syndrome in the subject of the present invention, the at least one symptom may be selected from the list of chronic diarrhea, constipation, bloating, gas, stomach cramps, skin problems, food sensitivity, nutritional deficiencies; aching, pain, confusion, difficulty concentrating, fatigue, chronic inflammation in any one or more parts of the body, and any combination thereof. In some embodiments, the at least one symptom is selected from the list of chronic diarrhea, constipation, bloating, gas, stomach cramps, acne, rash, eczema, food sensitivity, nutritional deficiencies, arthralgia, headache, confusion, difficulty concentrating, fatigue, chronic inflammation in any one or more parts of the body, and any combination thereof. In some embodiments, the at least one symptom is selected from the list of chronic diarrhea, constipation, bloating, gas, stomach cramps, chronic inflammation in any one or more parts of the body, and any combination thereof.

[0065] In some embodiments, at least one symptom is at least two symptoms selected from the list of chronic diarrhea, constipation, bloating, gas, stomach cramps, acne, rash, eczema, food sensitivity, malnutrition, arthralgia, headache, confusion, difficulty concentrating, fatigue, chronic inflammation of any one or more parts of the body, and any combination thereof. In some embodiments, at least one symptom is at least three symptoms selected from the list of chronic diarrhea, constipation, bloating, gas, stomach cramps, acne, rash, eczema, food sensitivity, malnutrition, arthralgia, headache, confusion, difficulty concentrating, fatigue, chronic inflammation of any one or more parts of the body, and any combination thereof.

[0066] At least one symptom is significantly improved, for example, by at least 5%, at least 10%, at least 20%, or at least 50%, compared to not using the method according to the present invention. The improvement can be assessed by a healthcare professional using any method known in the art, or by using a self-reported outcome scale of the subject.

[0067] In some embodiments, the present invention provides compounds of formula I or pharmaceutically acceptable salts or solvates thereof for use in methods of treating leaky gut, in which the subject of treatment is not suffering from celiac disease, IBD, IBS, ulcerative colitis, or Crohn's disease.

[0068] In some embodiments, the present invention provides compounds of formula I or pharmaceutically acceptable salts or solvates thereof for use in methods of treating leaky gut, where treatment is prevention.

[0069] In some embodiments, the present invention provides compounds of formula I or pharmaceutically acceptable salts or solvates thereof for use in methods of treating leaky gut, During the ceremony, X is selected from oxygen and sulfur. R is selected from the group including cyclohexyl, phenyl, and substituted phenyl; The substituted phenyl is substituted at any one or more positions with one to five identical or different substituents selected from the group consisting of fluoro; chloro; bromo; iodo, cyano, nitro, trifluoromethyl, C1-C6 linear and branched alkoxy, C1-C6 sulfoxy, -S-C1-C6 alkyl, C1-C6 linear and branched alkyl, alkenyl and alkynyl, C2-C6 linear and branched alkenyl, C2-C6 linear and branched alkynyl, C1-C6 fluoroalkyl, chloroalkyl, bromoalkyl, and iodoalkyl, COO-C1-C6 alkyl, O(CO)-C1-C6 alkyl, NH-C1-C6 alkyl, N(C1-C6 alkyl)2, CON(C1-C6 alkyl)2, and NH(CO)-C1-C6 alkyl; R' is selected from the group including hydrogen, C1-C6 linear and branched alkyl groups, and C3-C6 cycloalkyl groups, and R'' is selected from the group including hydrogen, benzyl, and acetyl.

[0070] In some embodiments, the present invention provides compounds of formula I or pharmaceutically acceptable salts or solvates thereof for use in a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome, wherein, X is selected from oxygen and sulfur. R is selected from the group including phenyl and substituted phenyl; The substituted phenyl is substituted at any one or more positions with one to five identical or different substituents selected from the group consisting of fluoro; chloro; bromo; iodo, cyano, nitro, trifluoromethyl, C1-C6 linear and branched alkoxy, C1-C6 sulfoxy, -S-C1-C6 alkyl, C1-C6 linear and branched alkyl, alkenyl and alkynyl, C2-C6 linear and branched alkenyl, C2-C6 linear and branched alkynyl, C1-C6 fluoroalkyl, chloroalkyl, bromoalkyl, and iodoalkyl, COO-C1-C6 alkyl, O(CO)-C1-C6 alkyl, NH-C1-C6 alkyl, N(C1-C6 alkyl)2, CON(C1-C6 alkyl)2, and NH(CO)-C1-C6 alkyl; R' is selected from the group including hydrogen, C1-C6 linear and branched alkyl groups, and C3-C6 cycloalkyl groups, and R'' is selected from the group including hydrogen, benzyl, and acetyl.

[0071] R may be selected from the group including phenyl and substituted phenyl, wherein the substituted phenyl is substituted at any one or more positions with one to five identical or different substituents selected from the group including fluoro; chloro; bromo; iodo; cyano; nitro; trifluoromethyl, C1-C6 linear and branched alkoxy; C1-C6 sulfoxy; -S-C1-C6 alkyl; C1-C6 linear and branched alkyl, alkenyl and alkynyl; C1-C6 fluoroalkyl, chloroalkyl, bromoalkyl and iodoalkyl; COO-C1-C6 alkyl, O(CO)-C1-C6 alkyl, NH-C1-C6 alkyl, N(C1-C6 alkyl)2, CON(C1-C6 alkyl)2, and NH(CO)-C1-C6 alkyl. Such a variation is represented as a1.

[0072] R may be selected from the group including phenyl and substituted phenyl, wherein the substituted phenyl is substituted at any one or more positions with one to five identical or different substituents selected from the group including fluoro; chloro; bromo; iodo; cyano; nitro; trifluoromethyl; C1-C6 linear and branched alkoxy; C1-C6 sulfoxy; -S-C1-C6 alkyl; C1-C6 linear and branched alkyl, alkenyl and alkynyl; and C1-C6 fluoroalkyl, chloroalkyl, bromoalkyl and iodoalkyl. Such a variation is represented as a2.

[0073] R may be selected from the group including phenyl and substituted phenyl, wherein the substituted phenyl is substituted at any one or more positions with 1 to 5 identical or different substituents selected from the group including fluoro; chloro; bromo; iodo; cyano; nitro; trifluoromethyl, NH-C1-C6 alkyl, N(C1-C6 alkyl)2, COO-C1-C6 alkyl, and O(CO)-C1-C6 alkyl. Such a variation is represented as a3.

[0074] R may be selected from the group including phenyl and substituted phenyl, wherein the substituted phenyl is substituted at any one or more positions with one to five identical or different substituents selected from the group including fluoro; chloro; bromo; iodo; C1-C6 linear and branched alkyl, alkenyl and alkynyl; NH-C1-C6 alkyl, N(C1-C6 alkyl)2, and C1-C6 linear and branched alkoxy. Such a variation is represented as a4.

[0075] R may be selected from the group including phenyl and substituted phenyl, wherein the substituted phenyl is substituted at any one or more positions with 1 to 5 identical or different substituents selected from the group including fluoro; chloro; bromo; iodo; C1-C3 linear and branched alkyl, alkenyl and alkynyl; COO-C1-C3 alkyl, and O(CO)-C1-C3 alkyl. Such a variation is represented as a5.

[0076] R may be selected from the group including phenyl and substituted phenyl, wherein the substituted phenyl is substituted at any one or more positions with one to five identical or different substituents selected from the group including C1-C6 linear and branched alkyl, alkenyl and alkynyl; and C1-C6 fluoroalkyl, chloroalkyl, bromoalkyl and iodoalkyl. Such a variation is represented as a6.

[0077] R may be phenyl. Such a variation is represented as a7.

[0078] The substituted phenyl may have one substituent at position 2, for example, position 3, or position 4. The phenyl ring may have two substituents at positions 2 and 6, for example, position 2 and 5, for example, position 2 and 3, for example, position 3 and 5, for example, position 2 and 4, for example, position 3 and 4. The phenyl ring may have three substituents at positions 2, 3 and 6, for example, position 2, 4 and 6, for example, position 2, 3 and 4, for example, position 2, 3 and 5, for example, position 3, 4 and 5. The phenyl ring may have four substituents at positions 2, 3, 4 and 6, for example, position 2, 3, 4 and 5, for example, position 2, 3, 5 and 6. The phenyl ring may have five substituents.

[0079] In all of the modifications a1 to a7, the group from which R is selected may further contain cyclohexyl.

[0080] In some variations, two of the substituents on the substituted phenyl are identical. In some variations, three of the substituents are identical. In some variations, four of the substituents are identical. In some variations, five of the substituents are identical. In other variations, all of the substituents are different.

[0081] R' may be selected from the group including hydrogen, C1-C6 linear and branched alkyl groups, and C3-C6 cycloalkyl groups. Such variations are represented as b1.

[0082] R' may be selected from the group including C3-C6 linear and branched alkyl groups, as well as C3-C6 cycloalkyl groups. Such variations are represented as b2.

[0083] R' may be selected from the group including hydrogen and C1-C3 linear and branched alkyl groups. Such modifications are represented as b3.

[0084] R' may be selected from methyl and ethyl. Such a variation is represented as b4.

[0085] R'' may be selected from the group including hydrogen, benzyl, and acetyl. Such a variant is represented as c1.

[0086] R'' may also be hydrogen. Such a variant is represented as c2.

[0087] X may be oxygen or sulfur. Such variations are represented as d1.

[0088] X may also be oxygen. Such a variation is represented as d2.

[0089] X may be sulfur. Such a variation is represented as d3.

[0090] It should be understood that each possible selection of X, R, R', and R'' groups disclosed herein is disclosed for use in any combination with any one or more other possible selections of X, R, R', and R'' groups disclosed herein.

[0091] Accordingly, in some embodiments, the present invention provides compounds of formula I or pharmaceutically acceptable salts or solvates thereof for use in a method of treating leaky gut, where the selection of X, R, R', and R'' is as listed below: a1+b1+c1+d1, a1+b2+c1+d1, a1+b3+c1+d1, a1+b4+c1+d1, a2+b1+c1+d1, a2+b2+c1+d1, a2+b3+c1+d1, a2+b4+c1+d1, a3+b1+c1+d1, a3+b2+c1+d1, a3+b 3+c1+d1、a3+b4+c1+d1、a4+b1+c1+d1、a4+b2+c1+d1、a4+b3+c1+d1、a4+b4+ c1+d1、a5+b1+c1+d1、a5+b2+c1+d1、a5+b3+c1+d1、a5+b4+c1+d1、a6+b1+c1+ d1, a6+b2+c1+d1, a6+b3+c1+d1, a6+b4+c1+d1, a7+b1+c1+d1, a7+b2+c1+d1, a7+b3+c1+d1, a7+b4+c1+d1, a1+b1+c2+d1, a1+b2+c2+d1, a1+b3+c2+d1, a 1+b4+c2+d1, a2+b1+c2+d1, a2+b2+c2+d1, a2+b3+c2+d1, a2+b4+c2+d1, a3+b1+c2+d1, a3+b2+c2+d1, a3+b3+c2+d1, a3+b4+c2+d1, a4+b1+c2+d1, a4+b2+ c2+d1、a4+b3+c2+d1、a4+b4+c2+d1、a5+b1+c2+d1、a5+b2+c2+d1、a5+b3+c2 +d1、a5+b4+c2+d1、a6+b1+c2+d1、a6+b2+c2+d1、a6+b3+c2+d1、a6+b4+c2+d 1. a7+b1+c2+d1, a7+b2+c2+d1, a7+b3+c2+d1, a7+b4+c2+d1, a1+b1+c1+d2, a1+b2+c1+d2, a1+b3+c1+d2, a1+b4+c1+d2, a2+b1+c1+d2, a2+b2+c1+d2, a2+ b3+c1+d2, a2+b4+c1+d2, a3+b1+c1+d2, a3+b2+c1+d2, a3+b3+c1+d2, a3+b4+c1+d2, a4+b1+c1+d2, a4+b2+c1+d2, a4+b3+c1+d2, a4+b4+c1+d2, a5+b1+c 1+d2, a5+b2+c1+d2, a5+b3+c1+d2, a5+b4+c1+d2, a6+b1+c1+d2, a6+b2+c1+d2, a6+b3+c1+d2, a6+b4+c1+d2, a7+b1+c1+d2, a7+b2+c1+d2, a7+b3+c1+d2a7+b4+c1+d2, a1+b1+c2+d2, a1+b2+c2+d2, a1+b3+c2+d2, a1+b4+c2+d2, a2+b1+c2+d2, a2+b2+c2+d2, a2+b3+c2+d2, a2+b4+c2+d2, a3+b1+c2+d2, a3+b 2+c2+d2, a3+b3+c2+d2, a3+b4+c2+d2, a4+b1+c2+d2, a4+b2+c2+d2, a4+b3+c2+d2, a4+b4+c2+d2, a5+b1+c2+d2, a5+b2+c2+d2, a5+b3+c2+d2, a5+b4+c2+ d2, a6+b1+c2+d2, a6+b2+c2+d2, a6+b3+c2+d2, a6+b4+c2+d2, a7+b1+c2+d2, a7+b2+c2+d2, a7+b3+c2+d2, a7+b4+c2+d2, a1+b1+c1+d3, a1+b2+c1+d3, a 1+b3+c1+d3、a1+b4+c1+d3、a2+b1+c1+d3、a2+b2+c1+d3、a2+b3+c1+d3、a2+ b4+c1+d3、a3+b1+c1+d3、a3+b2+c1+d3、a3+b3+c1+d3、a3+b4+c1+d3、a4+b1+ c1+d3、a4+b2+c1+d3、a4+b3+c1+d3、a4+b4+c1+d3、a5+b1+c1+d3、a5+b2+c1 +d3、a5+b3+c1+d3、a5+b4+c1+d3、a6+b1+c1+d3、a6+b2+c1+d3、a6+b3+c1+d 3、a6+b4+c1+d3、a7+b1+c1+d3、a7+b2+c1+d3、a7+b3+c1+d3、a7+b4+c1+d3、 a1+b1+c2+d3、a1+b2+c2+d3、a1+b3+c2+d3、a1+b4+c2+d3、a2+b1+c2+d3、a2+ b2+c2+d3、a2+b3+c2+d3、a2+b4+c2+d3、a3+b1+c2+d3、a3+b2+c2+d3、a3+b3 +c2+d3、a3+b4+c2+d3、a4+b1+c2+d3、a4+b2+c2+d3、a4+b3+c2+d3、a4+b4+c 2+d3、a5+b1+c2+d3、a5+b2+c2+d3、a5+b3+c2+d3、a5+b4+c2+d3、a6+b1+c2+ d3、a6+b2+c2+d3、a6+b3+c2+d3、a6+b4+c2+d3、a7+b1+c2+d3、a7+b2+c2+d3、a7+b3+c2+d3, a7+b4+c2+d3. ,

[0092] The letter-number combinations refer to the variants defined above, and as a result, for example, a1+b1+c1+d1, R is selected from the group including phenyl and substituted phenyl, and the substituted phenyl is at any one or more positions fluoro; chloro; bromo; iodo; cyano; nitro; trifluoromethyl; C1-C6 linear and branched alkoxy; C1-C6 sulfoxy; -S-C1-C6 alkyl; C1-C6 linear and branched alkyl, alkenyl and alkynyl; C1-C6 fluoroalkyl, chloroalkyl, bromoalkyl and iodoalkyl; COO-C1 The compounds are represented by being substituted with 1 to 5 identical or different substituents selected from the group including -C6 alkyl, O(CO)-C1-C6 alkyl, NH-C1-C6 alkyl, N(C1-C6 alkyl)2, CON(C1-C6 alkyl)2, and NH(CO)-C1-C6 alkyl (a1), where R' is selected from the group including hydrogen, C1-C6 linear and branched alkyl, and C3-C6 cycloalkyl (b1), where R'' is selected from the group including hydrogen, benzyl, and acetyl (c1), and where X is selected from oxygen and sulfur (d1).

[0093] In some embodiments, the present invention provides a compound of formula I or a pharmaceutically acceptable salt or solvate thereof for use in a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome, wherein the compound is capsaicin. In some embodiments, the present invention provides a compound of formula I or a pharmaceutically acceptable salt or solvate thereof for use in a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome, wherein the compound is thioamide capsaicin.

[0094] In preferred embodiments, the present invention provides a compound of formula I or a pharmaceutically acceptable salt or solvate thereof for use in a method of treating leaky gut, a method of treating leaky gut syndrome, or a method of improving at least one symptom associated with leaky gut syndrome, wherein R is a substituted phenyl having at least one substituent at the 4-position, the at least one substituent being selected from the group listed above for one of the options a1, a2, a3, a4, a5, or a6.

[0095] In other preferred embodiments, the present invention provides compounds of formula I or pharmaceutically acceptable salts or solvates thereof for use in a method of treating leaky gut, a method of treating leaky gut syndrome, or a method of improving at least one symptom associated with leaky gut syndrome, wherein R is a substituted phenyl having only one substituent, the substituent being at the 4-position and selected from the group listed above for one of the options a1, a2, a3, a4, a5, or a6.

[0096] In preferred embodiments, the present invention provides compounds of formula I or pharmaceutically acceptable salts or solvates thereof for use in a method of treating leaky gut, a method of treating leaky gut syndrome, or a method of improving at least one symptom associated with leaky gut syndrome, wherein X is oxygen; R is phenyl or a substituted phenyl as defined above (a1, a2, a3, a4, a5, a6, or a7); R' is methyl, ethyl, or isopropyl; and R'' is hydrogen, acetyl, or benzyl.

[0097] In other preferred embodiments, the present invention provides compounds of formula I or pharmaceutically acceptable salts or solvates thereof for use in a method of treating leaky gut, a method of treating leaky gut syndrome, or a method of improving at least one symptom associated with leaky gut syndrome, wherein X is oxygen; R is phenyl, 4-methylphenyl, 4-chlorophenyl, or 4-acetoxyphenyl; R' is methyl, ethyl, or isopropyl; and R'' is hydrogen, acetyl, or benzyl.

[0098] Preferred non-limiting examples of compounds for use in a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome according to the present invention include a list of compounds of formula I, where R, R', R'', and X are selected as follows: -R=phenyl; R'=methyl, R''=hydrogen, X=oxygen (phenylcapsaicin, V) -R=phenyl; R'=methyl, R''=acetyl, X=oxygen -R=phenyl; R'=methyl, R''=benzyl, X=oxygen -R=phenyl; R'=ethyl, R''=acetyl, X=oxygen -R=4-methylphenyl; R'=methyl, R''=acetyl, X=oxygen -R=4-chlorophenyl; R'=methyl, R''=acetyl, X=oxygen -R=4-methylphenyl; R'=methyl, R''=hydrogen, X=oxygen -R=4-acetoxyphenyl; R'=methyl, R''=hydrogen, X=oxygen -R=4-methylphenyl; R'=methyl, R''=benzyl, X=oxygen -R=4-acetoxyphenyl; R'=ethyl, R''=benzyl, X=oxygen -R=cyclohexyl; R'=methyl, R''=hydrogen, X=oxygen -R = cyclohexyl; R' = ethyl, R'' = hydrogen, X = oxygen

[0099] In some embodiments, the present invention provides compounds of formula I or pharmaceutically acceptable salts or solvates thereof for use in a method of treating leaky gut, a method of treating leaky gut syndrome, or a method of improving at least one symptom associated with leaky gut syndrome, wherein the compounds are selected from the group of phenyl-substituted 6-in derivatives of capsaicin, often called phenylcapsaicin(plural), where R is phenyl or substituted phenyl according to any of the above modifications a1 to a7.

[0100] In some embodiments, the present invention provides compounds of formula II or pharmaceutically acceptable salts or solvates thereof for use in a method of treating leaky gut, a method of treating leaky gut syndrome, or a method of improving at least one symptom associated with leaky gut syndrome, wherein R represents a phenyl or substituted phenyl selected from the above modifications a1 to a7. [ka]

[0101] In some embodiments, the present invention provides compounds of formula II or pharmaceutically acceptable salts or solvates thereof for use in a method of treating leaky gut, a method of treating leaky gut syndrome, or a method of improving at least one symptom associated with leaky gut syndrome, wherein R represents cyclohexyl or phenyl or substituted phenyl selected from the above modifications a1 to a7, and X is oxygen.

[0102] In some embodiments, the present invention provides compounds of formula II or pharmaceutically acceptable salts or solvates thereof for use in a method of treating leaky gut, a method of treating leaky gut syndrome, or a method of improving at least one symptom associated with leaky gut syndrome, wherein R represents a phenyl or substituted phenyl selected from the above modifications a1 to a7, and X is oxygen.

[0103] In some embodiments, the present invention provides compounds of formula II or pharmaceutically acceptable salts or solvates thereof for use in a method of treating leaky gut, a method of treating leaky gut syndrome, or a method of improving at least one symptom associated with leaky gut syndrome, wherein R represents a phenyl or substituted phenyl selected from the above modifications a1 to a7, and X is sulfur.

[0104] In some embodiments, the present invention provides a compound of formula III or a pharmaceutically acceptable salt or solvate thereof for use in a method of treating leaky gut, a method of treating leaky gut syndrome, or a method of improving at least one symptom associated with leaky gut syndrome, wherein X is selected from oxygen and sulfur (modification d1). [ka]

[0105] In some embodiments, the present invention provides compounds of formula IV or pharmaceutically acceptable salts or solvates thereof for use in a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome. [ka]

[0106] In some embodiments, the present invention provides phenylcapsaicin of formula V or a pharmaceutically acceptable salt or solvate thereof for use in a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome. [ka]

[0107] Alternatively, R', R'' and X are any combination of the bases outlined above (b1+c1+d1, b1+c2+d1, b1+c1+d2, b1+c2+d2, b1+c1+d3, b1+c2+d3, b2+c1+d1, b2+c2+d1, b2+c1+d2, b2+c2+d2, b2+c1+d3, b2+c2+d3, b3+c1+d1, b3+c2+d1, b3+c1+d2, R may be selected from b3+c2+d2, b3+c1+d3, b3+c2+d3, b4+c1+d1, b4+c2+d1, b4+c1+d2, b4+c2+d2, b4+c1+d3, b4+c2+d3), and R is selected from C1-C6 linear or branched alkyl groups, e.g., tert-butyl, e.g., cyclohexyl, e.g., cyclohexenyl, e.g., cyclohexadienyl.

[0108] The compounds of the present invention may contain one or more chiral centers and / or double bonds, and therefore may exist in different stereoisomeric forms such as double bond isomers (i.e., geometric isomers), enantiomers, and / or diastereomers. It should be understood that both stereoisomerically pure forms (e.g., geometrically pure, enantiomerically pure, or diastereomerically pure) and stereoisomeric mixtures are encompassed by the present invention. The present invention is considered to extend to diastereomers and enantiomers, as well as racemic mixtures. The compounds described herein can be divided into their geometric isomers, enantiomers, and / or diastereomers using methods known in the art.

[0109] Any of the compounds according to the present invention may be provided in the form of a prodrug. The term "prodrug" refers to a derivative of a pharmacologically active compound that, after administration, undergoes transformation such as metabolism in the body to release a pharmacologically active drug. Prodrugs may, but are not necessarily, be pharmacologically inactive until they are converted into an active compound. Prodrugs can be obtained by derivatizing one or more functional groups in an active compound with a pro group, i.e., a group that masks a functional group in the active compound and, under specific use conditions such as in vivo, is converted to release the functional group. The pro group should be nontoxic. A wide range of pro groups and methods for providing prodrugs are known to those skilled in the art.

[0110] Compounds for use in a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome according to the present invention may be present as active ingredients in desired dosage unit formulations, such as pharmaceutically acceptable compositions containing conventional pharmaceutically acceptable carriers.

[0111] Compositions for use in a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome according to the present invention are prepared in a substantially pure form from a compound of structure I according to the present invention. In some embodiments, the purity of the compound of structure I used to formulate the composition is at least about 95%, for example, at least 96%, 97%, 98%, or 99%. Preferably, the purity of the compound is at least 98%.

[0112] The composition may further comprise one or more of any conventional pharmaceutically acceptable excipients and / or carriers, such as solvents, fillers, diluents, binders, lubricants, flow enhancers, viscosity modifiers, surfactants, dispersants, disintegrants, emulsifiers, wetting agents, suspending agents, thickeners, buffers, pH adjusters, absorption retarders, stabilizers, antioxidants, preservatives, antifungal agents, chelating agents, adjuvants, sweeteners, fragrances, and colorants. The composition may be formulated using conventional formulation techniques known in the art, such as conventional mixing, dissolution, suspension, granulation, sugar-coated tablet manufacturing, polishing, emulsification, encapsulation, sealing, or compression processes.

[0113] In some embodiments, compositions for use in a method of treating leaky gut, a method of treating leaky gut syndrome, or a method of improving at least one symptom associated with leaky gut syndrome are formulated for oral and / or rectal and / or intraperitoneal administration.

[0114] The amount of the compound of structure I according to the present invention present in the composition may vary. In some embodiments, the amount of the compound according to the present invention present in the composition is 1 to 50% by weight, for example 1 to 30% by weight, for example 20 to 50% by weight. In other embodiments, the amount of the compound according to the present invention present in the composition is 30 to 70% by weight, for example 40 to 60% by weight. In yet another embodiment, the amount of the compound according to the present invention present in the composition is 50 to 100% by weight, for example 50 to 70% by weight, for example 50 to 80% by weight, for example 60 to 98% by weight, for example 70 to 95% by weight, for example 80 to 99% by weight, for example 95 to 100% by weight.

[0115] Furthermore, the compositions for use in the methods for treating leaky gut, leaky gut syndrome, or improving at least one symptom associated with leaky gut syndrome according to the present invention are substantially free of contaminants or impurities. In some embodiments, the level of contaminants or impurities other than the residual solvent in the composition is less than about 5% of the combined weight of the compound according to the present invention and other intended components. In certain embodiments, the level of contaminants or impurities other than the residual solvent in the composition is about 2% or 1% or less of the combined weight of the compound according to the present invention and other intended components.

[0116] In certain embodiments, the compounds or compositions used in the methods for treating leaky gut, treating leaky gut syndrome, or improving at least one symptom associated with leaky gut syndrome according to the present invention are sterile. Sterilization can be achieved by any suitable method, including but not limited to applying heat, chemicals, irradiation, high pressure, filtration, or a combination thereof.

[0117] In certain embodiments, compounds or compositions for use in a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome according to the present invention are formulated as pharmaceuticals, active pharmaceutical ingredients (APIs), food additives, dietary supplements, health supplements, nutritional supplements, over-the-counter supplements, medical foods, and / or pharmaceutical-grade supplements.

[0118] In certain embodiments, the method according to the present invention comprises the step of administering a compound or composition according to the present invention, the compound or composition being formulated as a pharmaceutical, a pharmacokinetic ingredient (API), a food additive, a dietary supplement, a health supplement, a nutritional supplement, an over-the-counter (OTC) supplement, a medical food, and / or a pharmaceutical-grade supplement.

[0119] Compounds or compositions for use in a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome according to the present invention are administered to a subject in a therapeutically effective dose, and the compounds or compositions include Proteobacteria, Actinobacteria, Firmicutes, Bacteroidetes, Bacteroides, Porphyromonas, and Bifidobacterium. It has no antibacterial effect or only a sublethal antibacterial effect against species such as *Lactobacillus*, *Clostridium*, *Prevotella*, *Ruminococcus*, *Alistipes*, *Dorea*, *Eubacterium*, *Faecalibacterium*, *Collinsella*, *Roseburia*, *Coprococcus*, and *Holdemania*.

[0120] The appropriate dosage may depend on the compound used, the stage of the condition, the patient's age and weight, etc., and can be routinely determined by an experienced practitioner in accordance with principles well known in the art. The appropriate daily dose of the compound according to the present invention may be in the range of about 0.001 mg / kg body weight to 1.0 mg / kg body weight. In some embodiments, the range is limited by the requirement that the dose should not have a lethal antimicrobial effect. For example, in some embodiments, the daily dose may be 0.001 to 0.01 mg / kg body weight, e.g., 0.001 to 0.005 mg / kg body weight, e.g., 0.003 to 0.008 mg / kg body weight, e.g., 0.005 to 0.01 mg / kg body weight. In other embodiments, the daily dose may be 0.005 to 0.05 mg / kg body weight, e.g., 0.005 to 0.02 mg / kg body weight, e.g., 0.008 to 0.05 mg / kg body weight. In other embodiments, the daily dose may be 0.01 to 0.1 mg / kg body weight, for example 0.01 to 0.05 mg / kg body weight, for example 0.03 to 0.08 mg / kg body weight, for example 0.05 to 0.1 mg / kg body weight. In other embodiments, the daily dose may be 0.05 to 0.5 mg / kg body weight, for example 0.05 to 0.2 mg / kg body weight, for example 0.08 to 0.5 mg / kg body weight. In yet another embodiment, the daily dose may be 0.1 to 1.0 mg / kg body weight, for example 0.3 to 0.8 mg / kg body weight, for example 0.5 to 1.0 mg / kg body weight.

[0121] The therapeutically effective dose of the compound according to the present invention can be administered as a single dose or in divided doses. The compound or composition according to the present invention may be administered once daily, twice or more daily, once every two days, once every three days, twice a week, or once a week, or as deemed appropriate by a healthcare professional. In certain embodiments, the compound or composition according to the present invention is administered once daily. In other embodiments, the compound or composition according to the present invention is administered twice daily. In some embodiments, the administration regimen is predetermined and is the same for the entire patient group. In other embodiments, the dose and frequency of treatment with the compound or composition according to the present invention are determined by a healthcare professional based on factors including, but not limited to, the stage of the disease, the severity of symptoms, the route of administration, the age of the subject, body weight, general health status, sex and / or diet, and / or the subject's response to treatment. When selecting an administration regimen, care should be taken to avoid lethal antimicrobial effects.

[0122] In some embodiments, the therapeutically effective dose is administered at regular intervals. In other embodiments, the dose is administered as needed or sporadically. The compounds or compositions according to the present invention may be administered by a medical professional or by self-administration. Depending on factors such as the formulation and route of administration, the compounds or compositions according to the present invention may be administered with or without food. In some embodiments, the compounds or compositions according to the present invention are administered at specific time intervals.

[0123] Compounds or compositions for use in a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome according to the present invention may be administered topically or systemically. Compounds or compositions according to the present invention may be administered by any route of administration, including but not limited to oral, intraperitoneal, sublingual, oral cavity, rectal, and enteral.

[0124] In some embodiments, the compound or composition is administered orally and / or rectally and / or intraperitoneally.

[0125] In preferred embodiments, the compound or composition is administered orally. In specific embodiments, the compound or composition is administered with or before a meal. In specific embodiments, the compound or composition is administered in a non-antimicrobial dose in a formulation that maximizes time in the intestines and slows the rate of drug metabolism.

[0126] In some embodiments, the compounds or compositions used in the methods for treating leaky gut, treating leaky gut syndrome, or improving at least one symptom associated with leaky gut syndrome according to the present invention are administered intraperitoneally, such as by intraperitoneal injection.

[0127] Preferred unit-dose formulations are formulations containing a therapeutically effective dose or appropriate fraction thereof of a compound of Structure I, as listed above, for use in a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome according to the present invention. Compositions for use in a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome may be provided in unit dosage forms as single doses in which all active and inactive components are combined in an appropriate system and the components do not need to be mixed before administration. Alternatively, the composition may be provided as a kit in which the drug, excipients and carriers are provided in two or more separate containers (e.g., ampoules, vials, tubes, bottles or syringes) and need to be combined to form the composition to be administered. The kit may contain one or more compounds or compositions according to the present invention and all other components in a unit dosage form or in two or more separate containers and may include instructions for storing, preparing, administering and / or using the composition.

[0128] In some embodiments, the duration of use of a compound or composition for use in a method of treating leaky gut, a method of treating leaky gut syndrome, or a method of improving at least one symptom associated with leaky gut syndrome according to the present invention is determined by the formulation used, and / or by the underlying mechanism of action detected and / or suspected in the subject being treated, and / or by the specific symptoms experienced by the subject being treated. In some embodiments, treatment is continued until further improvement is no longer expected based on the symptoms of the subject being treated. In certain embodiments, the duration of treatment with a compound or composition according to the present invention is at least 3 days, at least 1 week, at least 2 weeks, at least 1 month, at least 3 months, for example, 3 months, 6 months, 9 months. In some embodiments, treatment is stopped when the symptoms associated with leaky gut have subsided. In some embodiments, the duration is determined by a medical professional based on factors including, but not limited to, the nature and severity of the symptoms, the route of administration, the subject's age, weight, general health status, sex and / or diet, and / or the subject's response to treatment. In some embodiments, treatment is repeated when symptoms associated with leaky gut recur. In other embodiments, the compound or composition is administered chronically, such as in a continuous mode, in contrast to an acute mode, in order to maintain the initial therapeutic effect for a long period of time.

[0129] In certain embodiments, the compound or composition according to the present invention is administered alone. In other embodiments, the compound or composition according to the present invention is administered in combination with one or more other therapeutic agents. The one or more other therapeutic agents may be known to be effective against leaky gut and / or may have an additive or synergistic mechanism of action with the compound or composition according to the present invention for the treatment of leaky gut. The one or more other therapeutic agents may be known to be effective against other diseases or conditions related to and / or coexisting with leaky gut and / or may have an additive or synergistic mechanism of action with the compound or composition according to the present invention for the treatment of leaky gut. In some embodiments, the compound or composition according to the present invention is administered as part of a combination therapy. A combination therapy comprising the compound or composition according to the present invention may refer to a composition comprising the compound or composition according to the present invention in combination with one or more therapeutic agents, and / or the simultaneous administration of the compound or composition according to the present invention and one or more therapeutic agents, wherein the compound or composition according to the present invention and one or more other therapeutic agents are not formulated in the same composition. When using separate formulations, the compound or composition according to the present invention may be administered simultaneously with, intermittently, alternately with, before, after, or in combination thereof with the administration of another therapeutic agent. When using combination therapy, care should be taken to avoid lethal antibacterial effects.

[0130] Embodiments and features described in the context of one embodiment relating to a compound or composition for use in, for example, a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome are also applicable to other embodiments of the present invention.

[0131] In a further embodiment, the present invention provides a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome, comprising the step of administering an effective amount of a compound of formula I or any pharmaceutically acceptable salt or solvate thereof to a subject.

[0132] In some embodiments, the present invention provides a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome, comprising the step of administering to a subject a composition comprising an effective amount of a compound of formula I or any pharmaceutically acceptable salt or solvate thereof.

[0133] In some embodiments, the present invention provides a method for treating leaky gut, leaky gut syndrome, or increased intestinal permeability, comprising the step of administering an effective amount of a compound of formula I or a pharmaceutically acceptable salt or solvate thereof, or a composition comprising the compound of formula I, to the subject. During the ceremony, X is selected from oxygen and sulfur. R is selected from the group including cyclohexyl, phenyl, and substituted phenyl; The substituted phenyl is substituted at any one or more positions with one to five identical or different substituents selected from the group consisting of fluoro; chloro; bromo; iodo, cyano, nitro, trifluoromethyl, C1-C6 linear and branched alkoxy, C1-C6 sulfoxy, -S-C1-C6 alkyl, C1-C6 linear and branched alkyl, alkenyl and alkynyl, C2-C6 linear and branched alkenyl, C2-C6 linear and branched alkynyl, C1-C6 fluoroalkyl, chloroalkyl, bromoalkyl, and iodoalkyl, COO-C1-C6 alkyl, O(CO)-C1-C6 alkyl, NH-C1-C6 alkyl, N(C1-C6 alkyl)2, CON(C1-C6 alkyl)2, and NH(CO)-C1-C6 alkyl; R' is selected from the group including hydrogen, C1-C6 linear and branched alkyl groups, and C3-C6 cycloalkyl groups, and R'' is selected from the group including hydrogen, benzyl, and acetyl.

[0134] In some embodiments, the present invention provides a method for treating leaky gut, leaky gut syndrome, or increased intestinal permeability, comprising the step of administering an effective amount of a compound of formula I or a pharmaceutically acceptable salt or solvate thereof, or a composition comprising the compound of formula I, to the subject. During the ceremony, X is selected from oxygen and sulfur. R is selected from the group including phenyl and substituted phenyl; The substituted phenyl is substituted at any one or more positions with one to five identical or different substituents selected from the group consisting of fluoro; chloro; bromo; iodo, cyano, nitro, trifluoromethyl, C1-C6 linear and branched alkoxy, C1-C6 sulfoxy, -S-C1-C6 alkyl, C1-C6 linear and branched alkyl, alkenyl and alkynyl, C2-C6 linear and branched alkenyl, C2-C6 linear and branched alkynyl, C1-C6 fluoroalkyl, chloroalkyl, bromoalkyl, and iodoalkyl, COO-C1-C6 alkyl, O(CO)-C1-C6 alkyl, NH-C1-C6 alkyl, N(C1-C6 alkyl)2, CON(C1-C6 alkyl)2, and NH(CO)-C1-C6 alkyl; R' is selected from the group including hydrogen, C1-C6 linear and branched alkyl groups, and C3-C6 cycloalkyl groups, and R'' is selected from the group including hydrogen, benzyl, and acetyl.

[0135] In a further embodiment, the present invention provides a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome, comprising the step of administering an effective amount of phenylcapsaicin (V), or any pharmaceutically acceptable salt or solvate thereof, to a subject.

[0136] In some embodiments, the present invention provides a method for treating leaky gut, a method for treating leaky gut syndrome, or a method for improving at least one symptom associated with leaky gut syndrome, comprising the step of administering to a subject a composition comprising an effective amount of phenylcapsaicin (V) or any pharmaceutically acceptable salt or solvate thereof.

[0137] In a further embodiment, the present invention provides the use of a compound of formula I or any pharmaceutically acceptable salt or solvate thereof for inhibiting quorum sensing, inhibiting quorum sensing in the gut, treating leaky gut, treating leaky gut syndrome, or improving at least one symptom associated with leaky gut syndrome.

[0138] In a further embodiment, the present invention provides the use of a composition comprising an effective amount of a compound of formula I for inhibiting quorum sensing, inhibiting quorum sensing in the gut, treating leaky gut, treating leaky gut syndrome, or improving at least one symptom associated with leaky gut syndrome.

[0139] In a further embodiment, the present invention provides the use of phenylcapsaicin(V) or any pharmaceutically acceptable salt or solvate thereof for inhibiting quorum sensing, inhibiting quorum sensing in the gut, treating leaky gut, treating leaky gut syndrome, or improving at least one symptom associated with leaky gut syndrome.

[0140] In a further embodiment, the present invention provides the use of a composition comprising phenylcapsaicin (V) for inhibiting quorum sensing, inhibiting quorum sensing in the gut, treating leaky gut, treating leaky gut syndrome, or improving at least one symptom associated with leaky gut syndrome.

[0141] In further embodiments, the present invention provides the use of a compound of formula I or a pharmaceutically acceptable salt or solvate thereof, or a composition comprising the compound of formula I, for the inhibition of quorum sensing. The quorum sensing may be undesirable quorum sensing. The quorum sensing may occur in the intestines. The use may be therapeutic. The use may be non-therapeutic, for example, for the non-therapeutic prevention of biofilms.

[0142] The present invention is not limited to the embodiments and examples shown. While various embodiments of the disclosure are described herein, it will be apparent to those skilled in the art that such embodiments are provided merely as examples. Numerous modifications and changes to the embodiments described herein, as well as variations and substitutions thereof, will be apparent to those skilled in the art without departing from the disclosure. It should be understood that various alternative forms to the embodiments described herein may be used when practicing the disclosure.

[0143] It should be understood that all embodiments of this disclosure may be optionally combined with any one or more of the other embodiments described herein.

[0144] It should be understood that each component, compound, or parameter disclosed herein is disclosed for use alone or in combination with any one or more other components, compounds, or parameters disclosed herein. Furthermore, each amount / value or range of each component, compound, or parameter disclosed herein is disclosed in combination with any one or more other amounts / values ​​or ranges of each component, compound, or parameter disclosed herein. Therefore, it should be understood that any combination of amounts / values ​​or ranges of two or more components, compounds, or parameters disclosed herein is also disclosed in combination with each other for the purposes of this specification. All features described herein, and combinations of such features, are included within the scope of the invention, provided that the features do not conflict with each other.

[0145] It should be understood that each lower limit of each range disclosed herein should be interpreted as being disclosed in combination with each upper limit of each range disclosed herein for the same component, compound, or parameter. Therefore, a disclosure of two ranges should be interpreted as a disclosure of four ranges derived by combining each lower limit and each upper limit of each range. A disclosure of three ranges should be interpreted as a disclosure of nine ranges derived by combining each lower limit and each upper limit of each range, and so on. Furthermore, any specific amount / value of a component, compound, or parameter disclosed in the specification or examples should be interpreted as a disclosure of either a lower or upper limit of a range, and can therefore be combined with any other lower or upper limit or range or specific amount / value of the same component, compound, or parameter disclosed elsewhere in this application to form a range for that component, compound, or parameter. [Examples]

[0146] Example 1 - Inhibition of quorum sensing activity by phenylcapsaicin (V) using Chromobacterium violaceum CV026. Phenylcapsaicin (V) and capsaicin were assayed for their quorum sensing inhibitory activity using Chromobacterium violaceum CV026.

[0147] Chromobacterium violaceum CV026 is a violacein-negative double miniTn5 mutant derived from C. violaceum (ATCC31532) that was used to identify quorum sensing inhibition. Chromobacterium violaceum CV026 lacks self-inducible acylhomoserine lactone synthetase and therefore requires exogenous addition of N-hexanoylhomoserine lactone (C6-HSL) to undergo quorum sensing and produce a natural antibiotic called violacein, a water-insoluble purple pigment with antibacterial activity.

[0148] method: A stock solution of compound V and capsaicin at a concentration of 1000 ug was prepared by dissolving 1000 μg in 100 μl of ethanol and 900 μl of sterile water. The stock solution was successively diluted with twice the volume of solvent (10% ethanol aqueous solution) to obtain test solutions at concentrations of 500 / 250 / 125 / 62.5 / 31.25 / 15.625 / 7.81 μg / ml.

[0149] Anti-quorum sensing activity was measured using Chromobacterium violaceum CV026. The inducer N-hexanoyl homoserine lactone (C6-HSL) was purchased from Sigma-Aldrich. CV026 was cultured overnight under aerobic conditions at 30°C in Luria-Bertani (LB) medium (LB, 5g yeast extract, 10g tryptone, 5g NaCl, 1L water). Anti-quorum sensing activity of V and capsaicin was detected using a standard disk diffusion assay with a double-layer culture plate. 15ml of LB (1.0% agar) containing 50μl of C. violaceum CV026 with 20μg-1 kanamycin and 50ng / ml C6-AHL was overlaid on 15ml of LB medium (2.5% agar). Test concentrations of V and capsaicin were added to sterile discs (8 mm in diameter) and placed on agar. The plates were then incubated overnight at 30°C. Quorum sensing inhibition was detected as a less colored / colorless area around the disc where viable cells showing proliferation but quorum sensing inhibition were present. Measurements were taken from the outer edge of the disc to the edge of the anti-quorum sensing inhibition region. A formulation solvent (10% ethanol aqueous solution) was used as a negative control.

[0150] result: The biosensor strain C. violaceum CV026 is a mutant of the wild-type strain and is unable to produce its own AHL signal. It responds only to exogenous active signaling molecules, producing purple violacein. Therefore, the loss of this purple color around the disk when incubated with exogenous AHL indicates QS inhibition by the test compound. More specifically, the region of QS inhibition (mm beyond the disk) results in an opaque disk that is not purple but has bacterial growth. Inhibition of bacterial growth (antimicrobial activity) is indicated by a transparent inhibition region that does not contain bacteria. The results are shown in Table 1 below, where QS represents quorum sensing. [Table 1]

[0151] Discussion / Conclusion: In this CV026 bacterial quorum sensing inhibition assay, compound V was shown to be a dose-dependent quorum sensing inhibitor at doses lower than the observed sublethal antimicrobial dose. Specifically, compound V at doses of 61.5 / 31.25 / 15.625 / 7.81 μg / ml showed dose-responsive strong to weak quorum sensing inhibitory activity, but exhibited antimicrobial activity at higher doses (500 / 250 / 125 μg / ml).

[0152] The natural compound capsaicin did not exhibit quorum sensing inhibitory activity at non-antimicrobial doses (31.25 / 15.625 / 7.81 μg / ml), further reassuring the unexpected nature of the quorum sensing inhibitory activity of the compound for use in the method for treating leaky gut, the method for treating leaky gut syndrome, or the method for improving at least one symptom associated with leaky gut syndrome according to the present invention.

[0153] These results indicate that compound V, and by extension other compounds for use in the method for treating leaky gut, the method for treating leaky gut syndrome, or the method for improving at least one symptom associated with leaky gut syndrome according to the present invention, have a novel and unexpected mechanism of action at sublethal antimicrobial doses that reduce bacterial pathogenicity and thus improve GI inflammatory conditions such as leaky gut syndrome.

[0154] Example 2 - Investigation of the antibacterial effect of phenylcapsaicin (V) The antibacterial effects of phenylcapsaicin were investigated in vitro, and the bactericidal activity of phenylcapsaicin(V) against three bacterial pathogens was determined.

[0155] The tests included Salmonella, Campylobacter, and Listeria. Four different concentrations of phenylcapsaicin were tested: 19 ppm, 56 ppm, 167 ppm, and 500 ppm.

[0156] The results of the tests are shown in Figures 1, 2, and 3, clearly demonstrating that the non-lethal and sub-lethal doses of phenylcapsaicin and its derivatives can be experimentally determined and used to define and deliver effective doses of the compounds of the invention for treating leaky gut and leaky gut syndrome by inhibiting quorum sensing of intestinal bacteria.

[0157] Example 3 - Inhibition of biofilm formation using enteropathogenic Pseudomonas aeruginosa (ATCC® 15442) background: Quorum sensing is a regulatory mechanism used by many bacterial species to population-dependently control the expression of gene circuits. Quorum sensing uses small, diffusible molecules called autoinducers to monitor and mediate the size of bacterial populations. Autoinducers produced by bacteria diffuse and accumulate in the surrounding environment, and when they reach a threshold concentration, they diffuse back into the bacteria, many of which regulate the transcription of specific genes that could otherwise transform benign bacteria into pathogenic ones. Growing evidence indicates that bacterial quorum sensing is involved in regulating a variety of biological processes, including the gene expression of pathogenic factors, spore formation, biofilm formation, and motility, which often result in the symptoms of GI disorders commonly associated with leaky gut syndrome.

[0158] In commercially important Gram-negative bacteria exemplified in this assay using Pseudomonas aeruginosa, a common encapsulated Gram-negative rod-shaped bacterium, N-acylhomoserine lactone (AHL) is most commonly used as an inducer. The quorum sensing system can be interfered with in several ways. Inactivating the quorum sensing system of a bacterial pathogen can result in a significant reduction in the production of pathogenic factors. One measure of quorum sensing inhibition is the inhibition of biofilm formation by pathogenic bacteria, which is the focus of this assay.

[0159] the purpose: The objective of the study was to assay phenylcapsaicin (V) and capsaicin for their inhibition of biofilm formation using Pseudomonas aeruginosa (ATCC® 15442) bacteria.

[0160] method: A stock solution of 1000 ug of vitamin V and capsaicin was prepared by dissolving 1000 μg in 100 μl of ethanol and 900 μl of sterile water. The stock solution was successively diluted with twice the volume of solvent (10% ethanol aqueous solution) to obtain test solutions of 500 / 125 / 31.25 / 7.81 μg / ml.

[0161] The biofilm inhibitory activity was determined using Pseudomonas aeruginosa (ATCC® 15442). The inducer N-hexanoyl homoserine lactone (C6-HSL) was purchased from Sigma-Aldrich. ATCC® 15442 was cultured overnight at 30°C under aerobic conditions in Luria-Bertani (LB) medium (LB, 5g yeast extract, 10g tryptone, 5g NaCl, 1L water).

[0162] Crystal violet staining assay Biofilm formation was quantified using a modified method described by Stepanovic et al. (Journal of Microbiological Methods, 2000, 40(2), 175-19). A replica of a 96-well microplate was used, and the test solution was packed at the selected doses described above. The culture was grown in LB at 30°C for 18 hours and adjusted to an OD of 0.1 at 600 nm. 20 μL of culture was added to each well, and the remaining volume up to 200 μL per well was filled with LB. To prevent evaporation, unused external wells were filled with 200 μL of sterile water.

[0163] After incubation for 24 hours, the supernatant was discarded, and the wells were washed three times with 200 μL of sterile phosphate-buffered saline (PBS). The bacteria adhering to the wells were fixed with 200 μL of methanol for 15 minutes, the methanol was discarded, and the plate was dried at 25°C.

[0164] Each well was stained with 200 μL of 0.3% (w / v) crystal violet solution for 5 minutes at 25°C. Excess crystal violet was pipetted out, and the wells were rinsed five times with sterile PBS. 200 μL of 33% (v / v) glacial acetic acid was added to the wells for 30 minutes to solubilize the dye adhering to the cell walls. Optical density (OD) was measured at 595 nm using a microplate spectrophotometer as a direct measure of biofilm formation. Pseudomonas aeruginosa in 3% DMSO was used as a negative control.

[0165] result: Table 2 shows the effects of phenylcapsaicin and capsaicin on biofilm formation by Pseudomonas aeruginosa (ATCC® 15442), a bacterium with a well-characterized quorum sensing system. [Table 2]

[0166] Based on OD intensity obtained by crystal violet staining assay (CVSA), biofilms formed by Pseudomonas aeruginosa (ATCC® 15442) at 30°C were significantly reduced by phenylcapsaicin at concentrations below 125 ug / ml, although no dose-dependent reduction was observed. Capsaicin did not show a reduction in biofilm formation at the same concentrations.

[0167] Both compounds showed growth interference at a concentration of 500 ug / ml.

[0168] The biomass formed by the control at 30°C was sufficient for biofilm testing, but the biofilm at 37°C had an OD of approximately 2.9, which was deemed insufficient for quantifying biofilm reduction.

[0169] The control represents bacterial growth in a LB containing a 3% DMSO aqueous solution carrier blank.

[0170] The values ​​shown are the averages of a triple assay at OD 595 nm.

[0171] Values ​​with consecutive different characters are different with a 5% probability according to Tukey's test (P<0.05).

[0172] Discussion / Conclusion: In this biofilm inhibition assay, phenylcapsaicin demonstrated a potent inhibitor of biofilm formation in Pseudomonas aeruginosa (ATCC® 15442) bacteria. Capsaicin did not inhibit biofilm formation at any of the doses tested. These results suggest that phenylcapsaicin may have a novel and unexpected mechanism of action at sublethal antimicrobial doses that reduces bacterial pathogenicity and thus improve GI inflammatory conditions such as leaky gut syndrome.

[0173] Example 4 - Inhibition of biofilm formation using the pathogenic enterohemorrhagic bacterium Escherichia coli O157:H7. Based on the same background as in Example 3, the objective of this study was to assay phenylcapsaicin (V) and capsaicin for their inhibition of biofilm formation using the pathogenic enterohemorrhagic bacterium Escherichia coli O157:H7.

[0174] method: Stock solutions of phenylcapsaicin and capsaicin were prepared by dissolving 1000 μg of phenylcapsaicin in 100 μL of ethanol and 900 μL of sterile water. The stock solutions were successively diluted with twice the volume, and then four times the volume, of solvent (10% ethanol aqueous solution) to obtain test solutions of 500 / 125 / 31.25(32) / 7.81(8) μg / ml.

[0175] Biofilm inhibitory activity was determined using Escherichia coli (E. coli) O157:H7. The inducer N-hexanoylhomoserine lactone (C6-HSL) was purchased from Sigma-Aldrich. Escherichia coli (E. coli) O157:H7 was cultured overnight under aerobic conditions at 37°C in Luria-Bertani (LB) medium (LB, 5g yeast extract, 10g tryptone, 5g NaCl, 1L water).

[0176] Using Escherichia coli (E. coli) O157:H7 in 3% DMSO as a control, a crystal violet staining assay was performed in the same manner as in Experiment 3.

[0177] result: Table 3 shows the effects of phenylcapsaicin and capsaicin on biofilm formation by Escherichia coli (E. coli) O157:H7. [Table 3]

[0178] Based on the OD intensity obtained by the crystal violet staining assay (CVSA), biofilms formed by Escherichia coli (E. coli) O157:H7 at 30°C were significantly reduced by phenylcapsaicin at concentrations of 125 and 32 μg / ml, but the lowest dose tested, 8 μg / ml, had no effect.

[0179] Capsaicin did not show a reduction in biofilm formation at the same concentration.

[0180] Both compounds showed growth interference at a concentration of 500 μg / ml.

[0181] Similar to Pseudomonas aeruginosa (Example 3), the biomass formed by the control at 30°C was sufficient for biofilm testing. However, the biofilm at 37°C had an OD of approximately 2.9, which was deemed insufficient for quantifying biofilm reduction.

[0182] The control represents bacterial growth in a LB containing a 3% DMSO aqueous solution carrier blank.

[0183] The values ​​shown are the averages of a triple assay at OD 595 nm.

[0184] Values ​​with consecutive different characters are different with a 5% probability according to Tukey's test (P<0.05).

[0185] Discussion / Conclusion: In this biofilm inhibition assay, phenylcapsaicin was a good inhibitor of biofilm formation in Escherichia coli (E. coli) O157:H7 bacteria at intermediate concentrations of 125 and 32 μg / ml, but not at the lowest concentration tested of 8 μg / ml. Capsaicin did not inhibit biofilm formation at any of the doses tested. These results suggest that phenylcapsaicin continues to possess a novel and unexpected mechanism of action at sublethal antimicrobial doses that may reduce the pathogenicity of various bacteria and improve GI inflammatory conditions such as leaky gut syndrome.

[0186] Example 5 - Inhibition of biofilm formation using Salmonella enterica subsp. enterica serovar Paratyphi C (ATCC(registered trademark) BAA 1714(trademark)). Based on the same background as in Examples 3 and 4, the objective of this study was to assay phenylcapsaicin (V) and capsaicin for their inhibition of biofilm formation using the Salmonella enterica subsp. enterica serovar Paratyphi C (ATCC® BAA 1714®) bacterial strain.

[0187] method: Stock solutions of phenylcapsaicin and capsaicin were prepared by dissolving 1000 μg of phenylcapsaicin in 100 μl of ethanol and 900 μl of sterile water. The stock solutions were successively diluted with twice the volume, and then four times the volume, of solvent (10% ethanol aqueous solution) to obtain test solutions of 500 / 125 / 31.25(32) / 7.81(8) μg / ml.

[0188] The biofilm inhibitory activity of ATCC® BAA 1714® was determined. The inducer N-hexanoyl homoserine lactone (C6-HSL) was purchased from Sigma-Aldrich. ATCC® BAA 1714® was cultured overnight under aerobic conditions at 37°C in Luria-Bertani (LB) medium (LB, 5g yeast extract, 10g tryptone, 5g NaCl, 1L water).

[0189] A crystal violet staining assay was performed using ATCC(registered trademark) BAA 1714(trademark) in 3% DMSO as a control, similar to Experiment 3.

[0190] result: Table 4 shows the effects of phenylcapsaicin and capsaicin on biofilm formation by ATCC® BAA 1714®. [Table 4]

[0191] Based on the OD intensity obtained by the crystal violet staining assay (CVSA), biofilms formed by ATCC® BAA 1714® at 30°C were significantly reduced by phenylcapsaicin at concentrations of 32 μg / ml and the lowest dose tested, 8 μg / ml. Capsaicin did not show a reduction in biofilm formation at the same concentrations.

[0192] Both compounds showed growth interference at concentrations of 500 and 125 μg / ml.

[0193] Similar to the previous assay, the biomass formed by the control at 30°C was sufficient for the biofilm test. However, the biofilm at 37°C had an OD of approximately 2.6, which was deemed insufficient for quantifying biofilm reduction.

[0194] The control represents bacterial growth in a LB containing a 3% DMSO aqueous solution carrier blank.

[0195] The values ​​shown are the averages of a triple assay at OD 595 nm. (SD = -0.7)

[0196] Values ​​with consecutive different characters are different with a 5% probability according to Tukey's test (P<0.05).

[0197] Discussion / Conclusion: In this biofilm inhibition assay, phenylcapsaicin demonstrated good inhibition of biofilm formation in ATCC® BAA 1714® bacteria at low concentrations of 32 μg / ml and the lowest concentration tested at 8 μg / ml. Capsaicin did not inhibit biofilm formation at any of the doses tested. These results suggest that phenylcapsaicin continues to possess a novel and unexpected mechanism of action, even at sublethal antimicrobial doses, potentially reducing the pathogenicity of various bacteria and improving GI inflammatory conditions such as leaky gut syndrome.

[0198] Example 6 - Inhibition of biofilm formation using various capsaicin compounds with Salmonella enterica subsp. enterica serovar Paratyphi C (ATCC® BAA 1714®). Based on the same background as in Example 3, the objective of this study was to assay capsaicin, phenylcapsaicin (V), and three other capsaicins represented as a-c below for their inhibition of biofilm formation using the Salmonella enterica subsp. enterica serovar Paratyphi C (ATCC® BAA1714®) bacterial strain. [ka]

[0199] method: In Example 5, both phenylcapsaicin and capsaicin showed growth interference (antibacterial activity) at a concentration of 500 μg / ml, so this high proportion was excluded from the assay.

[0200] A stock solution of 500 μg of phenylcapsaicin, capsaicin, compound a, compound b, and compound c was prepared by dissolving 500 μg in 100 μl of ethanol and 900 μl of sterile water. The stock solution was successively diluted with four times the volume of solvent (10% ethanol aqueous solution) to obtain test solutions of 125 / 31.25(32) / 7.81(8) / 1.95(2) μg / ml.

[0201] The biofilm inhibitory activity of ATCC® BAA1714® was determined. The inducer N-hexanoyl homoserine lactone (C6-HSL) was purchased from Sigma-Aldrich. ATCC® BAA1714® was cultured overnight under aerobic conditions at 37°C in Luria-Bertani (LB) medium (LB, 5g yeast extract, 10g tryptone, 5g NaCl, 1L water).

[0202] A crystal violet staining assay was performed using ATCC(registered trademark)BAA1714(trademark) in 3% DMSO as a control, similar to Experiment 3.

[0203] result: Table 5 shows the effects of phenylcapsaicin, capsaicin, compound a, compound b, and compound c on biofilm formation by ATCC® BAA 1714®. [Table 5]

[0204] Based on the OD intensity obtained by the crystal violet staining assay (CVSA), the biofilm formed by ATCC® BAA1714® at 30°C was again significantly reduced by a new low dose of phenylcapsaicin at concentrations of 32 μg / ml, 8 μg / ml, and the lowest dose tested at 2 μg / ml. The 125 μg / ml dose still showed antimicrobial interference with the biofilm readings.

[0205] Capsaicin did not show a reduction in biofilm formation at any of the doses tested. The 125 μg / ml dose still showed antimicrobial interference with biofilm readings.

[0206] Phenylcapsacin analog a showed modest biofilm formation inhibitory activity at a dose of 2 μg / ml. Compounds b and c did not show any biofilm formation inhibitory activity.

[0207] Similar to the previous assay, the biomass formed by the control at 30°C was sufficient for the biofilm test. However, the biofilm at 37°C had an OD of approximately 2.6, which was deemed insufficient for quantifying biofilm reduction.

[0208] The control represents bacterial growth in a LB containing a 3% DMSO aqueous solution carrier blank.

[0209] The values ​​shown are the average of a triple assay at OD 595nm (SD+ / -0.66).

[0210] Values ​​with consecutive different characters are different with a 5% probability according to Tukey's test (P<0.05).

[0211] Discussion / Conclusion: In this biofilm formation inhibition assay, phenylcapsaicin was again demonstrated to be a good inhibitor of biofilm formation in ATCC® BAA1714® bacteria at concentrations of 32 μg / ml, 8 μg / ml, and a new, lower concentration of 2 μg / ml.

[0212] Of the three phenylcapsaicin analogs tested, only (a) showed somewhat modest biofilm formation inhibitory activity, close to 33% of the activity of phenylcapsaicin at a dose of 2 μg / ml. This result can be used to develop a QSAR model of biofilm inhibition by the phenylcapsaicin structure.

[0213] The results indicate that various capsaicins possess novel and unexpected mechanisms of action, potentially reducing the pathogenicity of various bacteria and improving GI inflammatory conditions such as leaky gut syndrome at sublethal antimicrobial doses. (1) Compounds of formula I, or pharmaceutically acceptable salts or solvates thereof, for use in methods of treating leaky gut (intestinal wall permeability): [ka] During the ceremony, X is selected from oxygen and sulfur. R is selected from the group including cyclohexyl, phenyl, and substituted phenyl; Substituting phenyl is fluoro; chloro; bromo; iodo, cyano, nitro, trifluoromethyl, C at any one or more positions. 1 -C 6 Linear and branched alkoxys, C 1 -C 6 Sulfoxy, -SC 1 -C 6 Alkyl, C 1 -C 6 Linear and branched alkyl, alkenyl and alkynyl, C 2 -C 6 Linear and branched alkenyls, C 2 -C 6 Linear and branched alkynyls, C 1 -C 6 Fluoroalkyl, chloroalkyl, bromoalkyl, and iodoalkyl, COO-C 1 -C 6 Alkyl, O(CO)-C 1 -C 6 Alkyl, NH-C 1 -C 6 Alkyl, N(C 1 -C 6 Alkyl) 2 CON(C 1 -C 6 Alkyl) 2 , and NH(CO)-C 1 -C 6 Substituted with 1 to 5 identical or different substituents selected from the group including alkyl groups; R' is hydrogen, C 1 -C 6 Linear and branched alkyl groups, and C 3 -C 6 Selected from the group including cycloalkyl, and R'' is selected from the group including hydrogen, benzyl, and acetyl. (2) A compound or a pharmaceutically acceptable salt or solvate thereof for use as described in (1), wherein R is selected from the group comprising phenyl and substituted phenyl. (3) A compound described in (1) or (2) or a pharmaceutically acceptable salt or solvate thereof for use in a method of treating asymptomatic increase in intestinal permeability in subjects experiencing one or more symptoms associated with leaky gut syndrome and showing a significant deviation in the level of at least one biomarker compared to the normal level of at least one biomarker associated with leaky gut, leaky gut syndrome and / or increased intestinal permeability. (4) A compound or a pharmaceutically acceptable salt or solvate thereof, as described in (1) or (2), for use as described in (1) or (3), wherein X is oxygen. (5) A compound or a pharmaceutically acceptable salt or solvate thereof, as described in (1) or (2), for use as described in (1) or (3), wherein X is sulfur. (6) A compound or a pharmaceutically acceptable salt or solvate thereof, as described in any one of items (1) to (5), for use as described in (1) or (3), wherein R' is methyl. (7) A compound or a pharmaceutically acceptable salt or solvate thereof, as described in any one of items (1) to (6), for use as described in (1) or (3), wherein R'' is hydrogen. (8) The compound has structure V:

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Claims

1. The compound phenylcapsaicin of formula V, 【Chemistry 1】 A composition for use in a method for treating leaky gut (intestinal wall leakage) by inhibiting bacterial quorum sensing, comprising a pharmaceutically acceptable salt or solvate thereof, The composition is administered to the subject in a sublethal antimicrobial dose. composition.

2. The composition according to claim 1, for use in a method for treating asymptomatic increase in intestinal permeability in a subject experiencing one or more symptoms associated with leaky gut syndrome and showing a significant deviation in the level of at least one biomarker compared to the normal level of at least one biomarker associated with leaky gut, leaky gut syndrome and / or increased intestinal permeability.

3. The composition according to claim 2, wherein at least one biomarker is zonulin.

4. The composition according to claim 2 or 3, wherein the subject has been diagnosed with leaky gut.

5. The composition according to any one of claims 2 to 4, wherein the subject has not been diagnosed with celiac disease, IBD, ulcerative colitis, or Crohn's disease.

6. The composition according to any one of claims 1 to 5, wherein the composition is administered to a subject by oral administration and / or intraperitoneal administration.

7. The composition according to any one of claims 1 to 6, wherein the composition is formulated for oral administration.

8. The composition according to any one of claims 1 to 7, wherein the composition is administered to the subject at a dose of 0.005 mg / kg body weight to 0.1 mg / kg body weight.

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