Use of compound for preparing drug for treating bacterial metabolism-related diseases

Abstract

A use of a compound as shown in structural formula I or a hydroquinone form, (C1-C4) quinone form, stereoisomer, hydrate, deuterated product, ester, solvate, metabolite, and pharmaceutically acceptable salt or prodrug thereof in preparation of a drug for treating bacterial metabolism-related diseases.

Description

Use of compounds for preparing drugs for treating bacterial metabolism-related diseases Technical Field

[0001] The present application relates to the field of biomedicine, and in particular to a compound that can be used to treat bacterial metabolism-related diseases, and to prepare a drug for treating bacterial metabolism-related diseases. Background Art

[0002] The intestinal microbiome is crucial to human health, participating in the metabolism and absorption of nutrients, while also defending against pathogens, inducing immunity, and maintaining a stable internal environment. Intestinal microbial imbalances are implicated in numerous diseases through the gut-liver and gut-brain axes, mediating their development and progression. These include metabolic dysfunction-associated steatohepatitis (MASH), obesity, diabetes, inflammatory bowel disease (IBD), and hepatic encephalopathy (HE), all of which have been the focus of recent research and development.

[0003] Gut dysbiosis is a pathogenic factor in metabolic dysfunction-associated fatty liver disease (MASLD) and promotes its progression to MASH by mediating inflammation, insulin resistance, bile acid, and choline metabolism. The gut microbiota can promote the development of hepatic steatosis through multiple mechanisms [Int. J. Mol. Sci. 22(2021)6485]. The role of the gut microbiota in the development of MASLD is as follows: (1) Microbial dysbiosis leads to increased intestinal ethanol production, which is toxic to the liver and can impair intestinal permeability by disrupting tight junctions; (2) Enteric pathogen-associated molecular patterns (PAMPs) such as LPS can bind to specific TLRs in the liver, thereby activating pro-inflammatory pathways that lead to liver inflammation and fibrosis; (3) The gut microbiota hydrolyzes choline to form dimethylamine and trimethylamine. Increased choline metabolism may lead to choline deficiency, thereby preventing the excretion of very low-density lipoprotein (VLDL) and triggering the accumulation of triglycerides in the liver; (4) Alterations in the intestinal microbiota may inhibit the secretion of fasting-induced adipokine (FIAF, also known as angiopoietin-related protein 4, ANGPTL4), a specific inhibitor of endothelial lipoprotein lipase (LPL), which releases triglycerides from very low-density lipoprotein particles into the liver. The net effect is to increase hepatic triglyceride storage; (5) Excessive short-chain fatty acids (SCFAs) are substrates for hepatic gluconeogenesis and lipogenesis, promoting the accumulation of hepatic free fatty acids (FFA) by inhibiting the activity of adenosine monophosphate-activated protein kinase (AMPK). Maintaining a balanced intestinal environment has been shown to be crucial for our health, and dysbiosis can lead to the development of MASLD / MASH through the gut-liver axis. Therefore, regulating the dysbiotic intestinal microbiota is an important therapeutic target to improve this disease, and improving the grade of MASLD can prevent its further progression to MASH.

[0004] The intestinal flora can promote the occurrence of obesity by regulating the growth and differentiation of adipocytes and affecting the metabolism and immune function of adipose tissue. In the intestinal flora of some obese people, the proportion of Firmicutes increases, while the proportion of Bacteroidetes decreases. This dysbiosis may affect energy recovery and fat storage. Secondly, the intestinal flora can affect fat and brain function through metabolites, thereby promoting the occurrence of obesity. Some short-chain fatty acids (SCFAs), such as acetate, propionate and butyrate, are metabolites produced by the metabolism of food by intestinal flora. For example, acetate, propionate and butyrate can affect the feeding behavior and energy metabolism of the hypothalamus by activating receptors such as GPR41 and GPR43, thereby affecting the occurrence of obesity. In addition, the intestinal flora can also affect appetite, food intake and energy metabolism by affecting the brain-gut axis, thereby affecting the occurrence of obesity.

[0005] Patients with type 2 diabetes experience moderate imbalance in the intestinal microecology, which causes changes in the species composition, abundance, and function of the intestinal flora, a decrease in probiotics, an increase in harmful bacteria, and a decrease in the species of flora, causing the body's inflammatory response level to increase, increase body mass index, promote insulin resistance, and ultimately lead to the occurrence of type 2 diabetes.

[0006] IBD is believed to be caused by a complex interplay of intestinal microbiota, host genes, immune system, and environmental factors. It is caused by genetic defects in host mucosal barrier function, intrinsic bacterial killing, or immune regulation, leading to continuous immune responses stimulated by pathogenic microbial antigens. In IBD patients, changes in microbial composition and function lead to increased immune stimulation, epithelial dysfunction, or enhanced mucosal permeability. Studies have found that in the intestinal inflammatory response of IBD patients, the number of macrophages increases significantly, and they can release a variety of bioactive substances such as interleukins (IL), transforming growth factors (TGF), and TNF, which play an important role in the occurrence and development of IBD. Antibiotics, as antibacterial substances active against bacteria, are widely used to treat IBD complications (bacteremia, abscesses, opportunistic and surgical site infections), etc. Commonly used antibiotics include metronidazole, ciprofloxacin, clofazimine, clarithromycin, rifaximin, etc. Several mechanisms mediate the therapeutic effects of antibiotics [Dig Dis Sci. 2020; 65(3), 757–788].

[0007] Oral gastrointestinal targeted antibiotics can modulate bacterial metabolism-related diseases through the following mechanisms: (1) Inhibition of pathogens. Each antibiotic has a unique antimicrobial spectrum, and most antibiotics inhibit pathogens and reduce overall bacterial diversity. (2) Increase in beneficial bacteria. Although many antibiotics reduce beneficial species, such as F. prausnitzii, some antibiotics can increase protective species. In particular, gut-targeted antibiotics such as rifaximin increase Lactobacillus, Bifidobacterium, and F. prausnitzii [J Antimicrob Chemother 2010; 65(12), 2556-65]. (3) Modification of bacterial metabolites. Changes in microbiota composition can alter microbial metabolites, increasing SCFAs and other beneficial products. (4) Immunomodulatory effects. Rifamycin, ciprofloxacin, metronidazole, and macrolides have mucosal immunomodulatory effects. For example, rifaximin is a gut-specific agonist of the human pregnane X receptor (PXR) that helps maintain mucosal homeostasis [Aliment Pharmacol Ther. 2016; 43(1), 27-36].

[0008] Therefore, oral gastrointestinal-targeted antibiotics offer a promising primary or adjunctive therapeutic strategy for modulating bacterial metabolism-related diseases, including metabolic dysfunction-associated steatohepatitis (MASH), obesity, diabetes, inflammatory bowel disease (IBD), irritable bowel syndrome with diarrhea (IBS-D), and / or hepatic encephalopathy (HE). Summary of the Invention

[0009] To achieve the above objectives and other related objectives, in one aspect, the present application provides a compound of structural formula I, or its hydroquinone form, (C1-C4) quinone form, stereoisomers, hydrates, deuterated forms, esters, solvates, metabolites, pharmaceutically acceptable salts or prodrugs, for use in the preparation of drugs for treating bacterial metabolism-related diseases.

[0010] Wherein, R is hydrogen or acetyl;

[0011] X is CH, CF, C-CN, C-CF3, C-Cl, C-Me, C-OMe, C-OCH2F, C-OCHF2 or N;

[0012] R i is (C1-C6)alkyl, optionally substituted (C3-C6)cycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl;

[0013] L is a linking group, and is selected from one or a combination of two to four of the following groups: a) (C1-C6) alkylene, b) (C1-C6) alkenylene, c) (C1-C6) alkynylene, d) (C3-C8) cycloalkylene, e) arylolefin, f) a divalent heterocyclic group containing 1 to 3 heteroatoms, wherein the heterocyclic group comprises a monocyclic, fused, spirocyclic or bridged ring, g) -C(=O)-, h) -C(=NOR x )-, where R x represents hydrogen, or optionally substituted (C1-C6) alkyl, i) -C(H)=N-, j) -O-, k) -S(O) n -, wherein n is an integer between 0 and 2, and l)-N(R y )-, where R y represents hydrogen, optionally substituted (C1-C6)alkyl,

[0014] Wherein, the carbon atom or nitrogen atom of the linking group L is optionally substituted by 1 to 3 substituents, and the substituents are selected from optionally substituted (C1-C6) alkyl, amino, (C1-C6) alkylamino, di(C1-C6) alkylamino, hydroxyl or (C1-C6) alkoxy.

[0015] In some embodiments, the L is selected from the following groups:

[0016] In some embodiments, the compound is any one of the following structural formulas:

[0017] In some embodiments, the bacterial metabolism-related disease comprises metabolic dysfunction-associated steatohepatitis (MASH), obesity, diabetes, inflammatory bowel disease (IBD), irritable bowel syndrome with diarrhea (IBS-D), and / or hepatic encephalopathy (HE).

[0018] On the other hand, the present application provides a method for treating bacterial metabolism-related diseases, comprising administering to a subject in need thereof an effective amount of the compound shown in structural formula I, or its hydroquinone form, (C1-C4) quinone form, stereoisomer, hydrate, deuterated form, ester, solvate, metabolite, pharmaceutically acceptable salt or prodrug.

[0019] In some embodiments, the bacterial metabolism-related disease comprises metabolic dysfunction-associated steatohepatitis (MASH), obesity, diabetes, inflammatory bowel disease (IBD), irritable bowel syndrome with diarrhea (IBS-D), and / or hepatic encephalopathy (HE).

[0020] The present application provides a gastrointestinal targeted antibiotic with a characteristic antibacterial spectrum, which has the following advantages: 1) An antibiotic with a selective antibacterial spectrum against intestinal conditional pathogens can reshape the stability of the intestinal flora by inhibiting intestinal conditional pathogens, reduce the entry of bacterial toxins such as LPS into the portal vein and the inflammatory response caused thereby, and reduce intestinal permeability; in addition, due to its low systemic exposure and high gastrointestinal exposure, it can reduce the occurrence of drug resistance and has few side effects; 2) It can act as a PXR agonist to regulate bile acid metabolism, regulate glycolipid metabolism, increase the excretion of harmful bacterial metabolites, and reduce inflammation; 3) It can act as a multifunctional small molecule that acts as an intestinal-specific FXR inhibitor (regulating bile acid metabolism, regulating glycolipid metabolism, and reducing weight). BRIEF DESCRIPTION OF THE DRAWINGS

[0021] The specific features of the inventions of this application are set forth in the appended claims. The features and advantages of the inventions of this application can be better understood by referring to the exemplary embodiments described in detail below and the accompanying drawings. A brief description of the drawings is as follows:

[0022] Figure 1 shows the comparison of aspartate aminotransferase (AST) levels among the compound-administered group described in the present application, the healthy control group, the model control group, and the obeticholic acid-administered group;

[0023] Figure 2 shows the comparison of plasma low-density lipoprotein LDL-C levels among the compound-administered group described in the present application, the healthy control group, the model control group, and the obeticholic acid-administered group;

[0024] Figure 3 shows the comparison of liver tissue degeneration scores between the compound-administered group described in the present application and the healthy control group, the model control group, and the obeticholic acid-administered group;

[0025] Figure 4 shows the comparison of pathological analysis results between the compound administration group described in the present application and the healthy control group, model control group, and obeticholic acid administration group;

[0026] Figure 5 shows the comparison of body weight changes of mice in the compound-administered group described in the present application, the healthy control group, the model control group, and the obeticholic acid-administered group after administration;

[0027] Figure 6 shows the comparison of the body weight change rate of mice in the compound administration group described in the present application, the healthy control group, the model control group, and the obeticholic acid administration group after administration. DETAILED DESCRIPTION

[0028] The following describes the implementation of the present invention through specific embodiments. People familiar with this technology can easily understand other advantages and effects of the present invention from the contents disclosed in this specification.

[0029] Definition of terms

[0030] In the present application, the term "compound" refers to the compound of the present application, and the term includes various pharmaceutically acceptable salts, hydrates or solvates of the compound of the present application. "Solvate" refers to an associate or complex of one or more solvent molecules and the compound of the present invention. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid and ethanolamine. The term "hydrate" refers to a complex in which the solvent molecule is water. Those skilled in the art will recognize that the term "prodrug" generally refers to the prodrug substance that can be metabolized to form a substance having the structure of the compound of the present application after administration to a subject; the term "metabolite" generally refers to the substance obtained by metabolism after administration to a subject of the compound of the present application, and the prodrugs and metabolites contained in such derivatives are included within the scope of the invention of the present application.

[0031] In the present application, the term "stereoisomer" generally refers to tautomers, mesomorphs, racemates, enantiomers, and / or diastereomers of the compounds of the present application. Wherein, the term "diastereomer" generally refers to stereoisomers with two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers can have different physical properties, for example, melting points, boiling points, spectral properties and reactivity. The terms "tautomer" or "tautomeric form" are used interchangeably and generally refer to structural isomers of different energies that can be mutually converted by low energy barriers. For example, proton tautomers (proton tautomers) (also referred to as prototropic tautomers) include mutual conversions performed by proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers (valence tautomers) include mutual conversions performed by the reorganization of some bonding electrons. The term "mesomorph" generally refers to a molecule containing an asymmetric atom, but due to its symmetry factors, the total optical rotation within the molecule is zero. The term "racemate" or "racemic mixture" generally refers to a composition consisting of equimolar amounts of two enantiomeric substances. Unless otherwise specified, all compounds appearing in this application are intended to include all possible optical isomers, such as single chiral compounds, or mixtures of various chiral compounds (i.e., racemates). Among all compounds in this application, each chiral carbon atom can optionally be in the R configuration or the S configuration, or a mixture of the R configuration and the S configuration.

[0032] In this application, the term "optionally substituted" generally means that the referenced group may be substituted or unsubstituted with one or more additional groups, the additional groups individually and independently selected from (but not limited to) alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heterocycloalkyl, hydroxy, alkoxy, mercapto, cyano, halogen, carbonyl, thiocarbonyl, isocyanato, thiocyanato, isothiocyanato, nitro, perhaloalkyl, perfluoroalkyl and amino including monosubstituted and disubstituted amino groups, and protected derivatives thereof. The position and number of such substituent groups are determined by the well-known valence limitations of each group.

[0033] In the present application, the term "alkyl" generally refers to a residue derived from an alkane by removing a hydrogen atom. An alkyl group can be substituted or non-substituted, substituted or non-substituted. The term "alkyl" generally refers to a saturated straight or branched aliphatic hydrocarbon group having a residue derived from the same carbon atom or two different carbon atoms of the parent alkane by removing a hydrogen atom, and can be a straight or branched group containing 1 to 20 carbon atoms, for example, a chain alkyl group containing 1 to 12 carbon atoms, for example, 1 to 6 carbon atoms. Non-limiting examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, propyl, butyl, etc. Alkyl groups may be substituted or unsubstituted, substituted or unsubstituted, e.g., when substituted, the substituents may be substituted at any available point of attachment, and the substituents may be independently selected from one or more substituents selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, and oxo. The term "alkylene" refers to a divalent alkyl group as defined above.

[0034] In this application, the term "alkenyl" generally refers to a straight or branched hydrocarbon group containing one or more double bonds. Illustrative examples of alkenyl include allyl, homoallyl, vinyl, crotyl, butenyl, pentenyl, and hexenyl. C 2-6 Illustrative examples of alkenyl groups include butadienyl, pentadienyl, hexadienyl and hexatrienyl and their branched forms. The position of the unsaturated bond (double bond) can be at any position of the carbon chain. The alkenyl group can be substituted or unsubstituted. The term "alkenylene" refers to a divalent hydrocarbon group derived from an alkenyl group as defined above.

[0035] In the present application, the term "alkynyl" generally refers to a partially unsaturated branched or straight chain hydrocarbon with at least one carbon-carbon triple bond. Alkynyl can be optionally substituted. In some embodiments, "C2-C3 alkynyl", "C2-C4 alkynyl", "C2-C5 alkynyl", "C2-C6 alkynyl", "C2-C7 alkynyl" and "C2-C8 alkynyl" generally refer to an alkynyl group containing at least 2, and at most 3, 4, 5, 6, 7 or 8 carbon atoms, respectively. Preferably, an alkynyl group is typically a C2-C6 alkynyl. In the present application, non-limiting examples of alkynyl groups include ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl etc. The term "alkynylene" generally refers to a partially unsaturated branched or straight chain divalent hydrocarbon group derived from an alkynyl as defined above.

[0036] In the present application, the term "cycloalkyl" generally refers to an "alkyl" comprising a cyclic form of a saturated monocyclic, bicyclic or polycyclic alkyl. The rings of the bicyclic or polycyclic systems can be fused together to connect via a single shared atom, i.e., they form a spirocyclic system or a bridged ring system. The cycloalkyl can contain 3 to 10 carbon atoms, such as 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms to form a ring. The example of a suitable cycloalkyl includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl. The example of suitable cycloalkyl includes but is not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, spiral [3,3] heptyl, spiral [3,4] octyl, spiral [4,3] octyl, bicyclo [4.1.0] heptyl, bicyclo [3.2.0] heptyl, bicyclo [2.2.1] heptyl, bicyclo [2.2.2] octyl, bicyclo [5.1.0] octyl or bicyclo [4.2.0] octyl. Cycloalkyl can be optionally substituted as described in the application. The term "cycloalkylidene" refers to a cycloalkyl group with two monovalent radical centers, and this monovalent radical center is obtained by removing a hydrogen atom from each of the two ring carbons.

[0037] In this application, the term "aryl" generally refers to a residue derived from an aromatic ring by removing a hydrogen atom. The term "aromatic ring" can refer to a 6- to 14-membered all-carbon monocyclic ring or a fused polycyclic ring (i.e., a ring sharing adjacent pairs of carbon atoms) with a conjugated π electron system, which can be 6- to 10-membered, such as benzene and naphthalene. The aromatic ring can be fused to a heteroaryl, heterocyclyl, or cycloalkyl ring, wherein the ring connected to the parent structure is the aryl ring. Aryl groups can be substituted or unsubstituted. When substituted, the substituents can be one or more of the following groups independently selected from the following groups: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio. Aryl groups can be optionally substituted as described herein.

[0038] As used herein, the term "heteroaryl" generally refers to a residue derived from a carbon atom in a heteroaromatic ring by removing a hydrogen atom. The term "heteroaromatic ring" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms may be selected from the group consisting of oxygen, sulfur, and nitrogen. Heteroaryl groups may be 5 to 10-membered, and may be 5- or 6-membered, such as furanyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, and the like. The heteroaryl ring may be fused to an aryl, heterocyclyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the heteroaryl ring. Heteroaryl can be optionally substituted or unsubstituted. When substituted, the substituent can be one or more of the following groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio. Heteroaryl can be optionally substituted as described herein.

[0039] In this application, as known to those skilled in the art, terms such as "alkyl", "alkenyl", "cycloalkyl", etc. may be preceded by an identifier to indicate the number of atoms present in the group in a particular case, for example, C1-C4 alkyl, C3-C7 cycloalkyloxy, C1-C4 alkylcarbonylamino, etc., and the subscript number after "C" indicates the number of carbon atoms present in the group. For example, C3 alkyl refers to an alkyl group with three carbon atoms (e.g., n-propyl, isopropyl); C 1-10 In the examples, the members of the group may have any number of carbon atoms falling within the range of 1-10.

[0040] As used herein, the term "heterocyclic group" generally refers to a monocyclic, bicyclic, or tricyclic, saturated or partially unsaturated, non-aromatic ring system having 3 to 20 ring atoms, including systems containing fused rings, in which at least one ring atom is a heteroatom. Examples of heteroatoms include nitrogen, oxygen, and sulfur. In some embodiments, heterocyclic groups refer to saturated ring systems, such as 3- to 12-membered saturated heterocyclic ring systems, or 3- to 8-membered saturated heterocyclic ring systems. In some embodiments, heterocyclic groups refer to 5- to 8-membered saturated heterocyclic ring systems. In some embodiments, heterocyclic groups refer to 5- to 6-membered saturated heterocyclic ring systems. In some embodiments, heterocyclic groups include 1 to 4 heteroatoms. In some embodiments, heterocyclic groups include 3- to 7-membered monocyclic rings having one or more heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, heterocyclic groups include 4- to 6-membered monocyclic rings having one or more heteroatoms selected from nitrogen, oxygen, and sulfur. In another example, heterocyclic groups include 3-membered monocyclic rings. In another example, heterocyclic groups include 4-membered monocyclic rings. In another example, the heterocyclyl group includes a 5-6 membered monocyclic ring. In one example, the heterocyclyl group includes 0 to 3 double bonds.

[0041] As used herein, the term "annular ring" generally refers to two or more rings sharing a pair of adjacent carbon atoms, forming a larger ring structure. In this structure, the shared carbon atoms belong to both rings. A common example of annular ring is naphthalene, which has two benzene rings sharing a pair of carbon atoms.

[0042] As used herein, the term "spirocycle" generally refers to a structure in which two rings are connected by a common carbon atom, called a spiro atom, which is typically a tetravalent carbon atom. This structure gives the two rings a propeller-like structure, and they are not in the same plane. A common example of a spirocycle is spiro[4.5]decane, which has two rings of different sizes connected by a shared carbon atom.

[0043] In this application, the term "bridged ring" generally refers to a ring structure consisting of three or more rings, and these rings are connected by two non-adjacent atoms (usually carbon atoms) to form a bridge-like ring structure. Among them, the two atoms are bridgehead atoms, and what connects the two bridgehead atoms is a bridging atom or a bridging chain. A common example of a bridged ring structure is a bridged ring terpene, such as pinane. In these structures, the bridge does not share adjacent atoms like a ring, but connects atoms farther away to form a new ring.

[0044] In this application, the term "subject in need" generally includes human patients and other mammalian subjects receiving prophylactic or therapeutic treatment, including but not limited to non-human primates, laboratory animals such as rabbits, dogs, rats and mice, and other animals. These "subjects in need" can be, for example, subjects with a confirmed specific disease, subjects receiving treatment related to a specific disease, subjects with a tendency or risk of developing a specific disease, etc.

[0045] In this application, the term "administer" generally refers to introducing the drug or a composition comprising the drug into the patient's body by any introduction or delivery route. Any method known to those skilled in the art for contacting cells, organs or tissues with the drug or its composition can be used.

[0046] As used herein, the term "effective amount" generally refers to an amount sufficient to achieve or at least partially achieve the desired therapeutic effect. An "effective amount" of a drug or therapeutic agent generally refers to an amount sufficient to cure or at least partially prevent the disease and its complications in a patient already suffering from the disease. The effective amount for this purpose will depend on the severity of the infection and the overall state of the patient's own immune system.

[0047] Without intending to be bound by any theory, the following examples are merely for illustrating the compounds and uses of the present application, and are not intended to limit the scope of the present invention.

[0048] Example

[0049] All compounds provided herein can be identified and prepared by following the protocols disclosed in US Pat. No. 7,247,634, which is incorporated herein by reference in its entirety.

[0050] Example 1

[0051] Antibacterial activity of the compounds of the present application against specific strains

[0052] 1. Strains: As shown in Table 1, all strains were obtained from the American Type Culture Collection (ATCC) and stored frozen at -80°C. Two days prior to antimicrobial activity testing, strains were revived. A small amount of the frozen bacteria was scraped with a sterile inoculating loop and streaked onto solid culture medium (TSA + 5% defibrinated sheep blood). The culture was then incubated at 35±2°C in a suitable atmosphere for 20-48 hours. A single pure colony was then picked from the resuscitation medium with a sterile inoculating loop, streaked onto solid culture medium again, and incubated under suitable conditions for 20-48 hours.

[0053] Table 1 Sources of strains

[0054] 2. Preparation of Bacterial Suspension

[0055] Pick 5-10 bacterial colonies from the solid culture medium and resuspend them in 5 mL of normal saline. Adjust the bacterial suspension to 0.5 McFarland (1-2×10 8 CFU / ml), Klebsiella pneumoniae, Escherichia coli, Shigella and Lactobacillus jensenii were diluted 100 times (~10 6 CFU / mL, anaerobic Peptostreptococci, Ruminococci, Eggerella tarda and Clostridium difficile were diluted 10 times with normal saline (~10 7 CFU / mL) and set aside.

[0056] 3. Broth Dilution Method MIC Determination

[0057] The test compounds were dissolved in DMSO and prepared into a 5120 μg / mL stock solution. The compounds were serially diluted to prepare a series of compound dilution working solutions (twice the final test concentration). 50 μL of the series of compound dilution working solutions were pipetted into the corresponding 96-well plates using a dispenser. 50 μL of the diluted bacterial suspension was added to each well, and the inoculum size was approximately 5 × 10 4 CFU / well. Read the number after 20-24 hours of incubation according to the bacterial culture conditions in the table above.

[0058] 4. Agar Dilution Method MIC Determination

[0059] The test compounds were dissolved in DMSO and the compounds were serially diluted to prepare a series of compound dilutions (100 times the final test concentration). Columbia culture medium containing 5% defibrinated sheep blood was used for anaerobic bacterial susceptibility testing. All culture media were prepared and stored according to the agar method of CLSIM07. 200 μL of compound dilution was added to the melted Columbia blood agar balanced in a water bath at 45°C to 50°C. The agar and drug solution were thoroughly mixed and poured onto a horizontal surface. The plate was allowed to solidify and then used. 3 μL of bacterial suspension was drawn up using a spray gun and inoculated into the compound dilution plate. The final inoculation volume was approximately 10 4 The inoculated agar plates were placed in an anaerobic box containing a GasPak bag and incubated at 35-37°C for 46-48 hours before reading. The results are shown in Table 2.

[0060] Table 2 Minimum inhibitory concentration (MIC) results of test compounds against specific strains (unit: μg / mL)

[0061] According to the results in Table 2, the compounds of the present application have high antibacterial activity against common intestinal conditional pathogens, which is better than rifaximin (such as Klebsiella pneumoniae, Escherichia coli and Shigella) or has similar antibacterial activity against intestinal conditional pathogens. The selectivity for probiotics (Lactobacillus jensenii) is significantly better than rifaximin. The compounds of the present application can reshape the stability of the intestinal flora by inhibiting intestinal conditional pathogens.

[0062] Example 2

[0063] Antibacterial activity of the compounds of the present application against specific strains

[0064] 1. Strains: As shown in Table 3, all strains were obtained from the American Type Culture Collection (ATCC) and the China Industrial Culture Collection (CICC) and stored frozen at -80°C. Two days prior to antimicrobial activity testing, strains were revived. A small amount of the frozen bacteria was scraped with a sterile inoculating loop and streaked onto solid culture medium (TSA + 5% defibrinated sheep blood). The culture was then incubated at 35 ± 2°C in a suitable atmosphere for 20-48 hours. A single pure colony was then picked from the resuscitation medium with a sterile inoculating loop, streaked onto solid culture medium again, and incubated under suitable conditions for 20-48 hours.

[0065] Table 3

[0066] 2. Preparation of Bacterial Suspension

[0067] Pick 5-10 bacterial colonies from the solid culture medium and resuspend them in 5 mL of normal saline. Adjust the bacterial suspension to 0.5 McFarland (1-2×10 8 CFU / ml, and diluted 10-fold with normal saline (~10 7 CFU / mL) and set aside.

[0068] 3. Agar Dilution Method MIC Determination

[0069] The test compounds were dissolved in DMSO, and the compounds were serially diluted to prepare a series of compound dilutions (100 times the final test concentration). Columbia culture medium containing 5% defibrinated sheep blood was used for drug sensitivity testing of anaerobic bacteria. All culture media were prepared and stored according to the agar method of CLSIM07. 200 μL of compound dilution was added to 20 mL of melted Columbia blood agar balanced in a 45°C to 50°C water bath. The agar and drug solution were thoroughly mixed and poured onto a horizontal surface. The plate was allowed to solidify and then used. 3 μL of bacterial suspension was drawn up using a spray gun and inoculated into the compound dilution plate. The final inoculation volume was approximately 10 4 The inoculated agar plates were placed in an anaerobic box containing a GasPak bag and incubated at 35-37°C for 46-48 hours before reading. The results are shown in Table 4.

[0070] Table 4 Minimum inhibitory concentration (MIC) results of test compounds against specific strains (unit: μg / mL)

[0071] According to the results in Table 4, the compounds of the present application have antibacterial activity against specific BSH-related strains. Except for Clostridium perfringens, most BSH-related strains are probiotics. The compounds of the present application can selectively inhibit intestinal conditional pathogens and have little effect on probiotics. It is estimated that they will not cause an imbalance of the intestinal flora or the possibility of causing an imbalance of the intestinal flora is low.

[0072] Example 3

[0073] PXR agonist activity assay of the compounds of the present application

[0074] In this example, PXR receptor agonist activity was measured using a HepG2 cell line transfected with PXR. The procedure was as follows: 1) On day 1, HepG2 cells were digested and plated onto 6 cm dishes; 2) On day 2, HepG2 cells were transfected with the pBIND-PXR plasmid and the PGL4.35 plasmid using JetPRIME transfection reagent; 3) On day 3, transfected cells were harvested, resuspended in phenol red-free DMEM medium containing 5% charcoal-stripped serum, counted, and plated into 96-well plates. Test compound (10 μg / mL), positive compound (10 μg / mL), or blank wells (DMSO) were added, and the plates were incubated in an incubator; 4) On day 4, 22 hours after drug addition, Bright-Glo Reagent was added to the experimental wells according to the Bright-Glo Luciferase Assay System instructions. After shaking and lysis, the luminescence signal was read using a multi-microplate reader. The results are shown in Table 5.

[0075] Table 5 PXR agonist activity of test compounds

[0076] According to the results in Table 5, the compounds of the present application have PXR agonist activity superior to that of rifampicin and rifaximin, and can act as PXR agonists to regulate bile acid metabolism and glycolipid metabolism, increase the excretion of bacterial toxins or harmful bacterial metabolites, and reduce inflammation.

[0077] Example 4

[0078] Pharmacokinetic study of the compound of the present application in mice

[0079] In this example, ICR mice were administered intravenously (IV, 5 mg / kg) and orally (PO, 50 mg / kg). Plasma samples (anticoagulated with EDTA-K2) were collected before administration (0 hour) and at 0.25 (15 minutes), 0.5, 1, 2, 4, 6, 10, and 24 hours after administration. The concentration of the test compound in plasma was determined by high-performance liquid chromatography-tandem mass spectrometry (LC-MS / MS), and the pharmacokinetic parameters of the compound were calculated to investigate the pharmacokinetic characteristics of the compound in ICR mice. The results are shown in Table 6.

[0080] Table 6 Pharmacokinetic study of compounds in ICR mice

[0081] NR stands for Not Applicable.

[0082] The results in Table 6 indicate that the compounds of the present application have low systemic exposure, extremely low bioavailability, and gastrointestinal targeting, which can reduce safety risks associated with higher systemic exposure, minimize side effects, and reduce the development of drug resistance. Furthermore, due to their gastrointestinal targeting, although the test compound has strong PXR agonist activity, it does not cause drug interactions like systemically administered rifampicin.

[0083] Example 5

[0084] Efficacy test of the compound of the present application in the HFD model of mice

[0085] In this embodiment, C57BL / 6J mice were fed a high-fat diet for about 12 weeks for MALFD mouse modeling. At this time, mice had the characteristics of weight gain, alanine aminotransferase (ALT elevation), aspartate aminotransferase (AST) relative to conventionally fed mice. After about 12 weeks of high-fat feeding, they were randomly divided into groups of 8, each group of 8, divided into a model control group (high-fat feeding solvent group), a positive control group (obeticholic acid, high-fat feeding) and a compound administration group (high-fat feeding), and 8 normal mice were selected to enter a healthy control group (conventionally fed solvent group). Each group of animals was gavaged once a day to give solvent or test compound. After 28 days of continuous administration, the weight of each group of mice was measured, and weight changes and serum samples and liver tissue pathology analysis were recorded. The group design and dosage regimen are shown in Table 7.

[0086] Table 7

[0087] The results of aspartate aminotransferase (AST) and plasma low-density lipoprotein (LDL-C) levels in the test compound-treated groups, the healthy control group, the model control group, and the obeticholic acid-treated groups are shown in Figures 1 and 2. The results of liver tissue degeneration score (Steatosis score) and pathology (NAS score) analysis are shown in Figures 3 and 4. The weight changes and weight change rates of mice after drug administration are shown in Figures 5 and 6. The results showed that the test compounds had significant effects on lowering LDL-C, liver tissue degeneration score, pathological NAS score, and body weight, indicating that this series of compounds has the ability to regulate bacterial metabolism and provides a promising primary or adjuvant treatment strategy for diseases related to bacterial metabolic dysfunction.

Claims

1. Use of a compound represented by Structural Formula I, or its hydroquinone form, (C1-C4) quinone form, stereoisomer, hydrate, deuterated compound, ester, solvate, metabolite, pharmaceutically acceptable salt or prodrug, in the preparation of a medicament for treating diseases related to bacterial metabolism, Among them, R is hydrogen or acetyl; X is C-H, C-F, C-CN, C-CF3, C-Cl, C-Me, C-OMe, C-OCH2F, C-OCHF2 or N; R i is a (C1-C6) alkyl, an optionally substituted (C3-C6) cycloalkyl, an optionally substituted aryl, or an optionally substituted heteroaryl; L is a linking group and is selected from one or a combination of two to four of the following groups: a) (C1-C6) alkylene, b) (C1-C6) alkenylene, c) (C1-C6) alkynylene, d) (C3-C8) cycloalkylene, e) arylene, f) a divalent heterocyclic group containing one to three heteroatoms, the heterocycle comprising a monocyclic, fused-ring, spiro or bridged ring, g) -C(=O)-, h)-C(=N-O-R x )- wherein R x represents hydrogen, or an optionally substituted (C1-C6) alkyl group, i) -C(H)=N-, j) -O-, k)-S(O) n -, where n is an integer between 0 and 2, and l)-N(R y )-, wherein R y represents hydrogen, optionally substituted (C1-C6) alkyl, wherein the carbon atom or nitrogen atom of the linking group L is optionally substituted with one to three substituents selected from optionally substituted (C1-C6) alkyl, amino, (C1-C6) alkylamino, di(C1-C6) alkylamino, hydroxy or (C1-C6) alkoxy.

2. According to the use described in claim 1, L is selected from the following groups:

3. The use according to claim 1 or 2, wherein the compound is any one of the following structural formulas:

4. The use according to any one of claims 1-3, wherein the bacterial metabolism-related disease comprises metabolic dysfunction-associated steatohepatitis (MASH), obesity, diabetes, inflammatory bowel disease (IBD), diarrhea-predominant irritable bowel syndrome (IBS-D), and / or hepatic encephalopathy (HE).

5. A method for treating a bacterial metabolism-related disease, comprising administering to a subject in need thereof an effective amount of the compound of formula I according to any one of claims 1-3, or its hydroquinone form, (C1-C4) quinone form, stereoisomer, hydrate, deuterated compound, ester, solvate, metabolite, pharmaceutically acceptable salt or prodrug.

6. The method according to claim 5, wherein the bacterial metabolism-related disease comprises metabolic dysfunction-associated steatohepatitis (MASH), obesity, diabetes, inflammatory bowel disease (IBD), diarrhea-predominant irritable bowel syndrome (IBS-D), and / or hepatic encephalopathy (HE).

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