Preparation method and use of a benzimidazole-2-one derivative

Abstract

The present application provides a compound of formula (I) or a pharmaceutically acceptable salt thereof. A method for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided. The use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a CYP1A1 inhibitor is provided. The use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or preventing tumors, bacterial infections, sepsis, inflammation, or organ damage is provided.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly to benzimidazole-2-one derivatives, a preparation method thereof, and their use as CYP1A1 enzyme inhibitors or antibacterial drugs. Background Art

[0002] The cytochrome P450 (CYP) 1 family of enzymes mainly consists of several enzymes such as CYP1A1, CYP1A2, and CYP1B1. Among them, CYP1A1 is a monooxygenase mainly present in the lungs, gastrointestinal tract, skin, and other extrahepatic tissues, mainly mediating oxidation-related reactions such as hydroxylation, epoxidation, and O-demethylation, and having a wide range of physiological functions. CYP1A1 is mainly involved in the metabolic degradation of exogenous harmful substances such as dioxins and aflatoxins, and is also a key enzyme for the metabolism of various drugs and hormones. As an important metabolic enzyme in the body, CYP1A1 is also involved in various physiological processes of the body, such as immune response and oxidative stress.

[0003] However, the activity of CYP1A1 is closely related to the occurrence and development of various diseases, including but not limited to tumors, bacterial infections, sepsis, inflammation (acute and chronic), organ damage (such as oxidative stress damage), etc.

[0004] The ability of CYP1A1 to catalyze the metabolism of various exogenous and endogenous compounds (including polycyclic aromatic hydrocarbons) into carcinogenic derivatives has been widely studied. It is mainly involved in the metabolism of polycyclic aromatic hydrocarbon compounds (PAHs), metabolizing them into epoxides, which play an important role in carcinogenesis and mutagenesis. This process is closely related to tumor progression and the occurrence of various types of cancers, including liver cancer, lung cancer, breast cancer, prostate cancer, breast cancer, etc. Under special circumstances, after exogenous substances bind to the aryl hydrocarbon receptor (AhR) in the body as ligands, it leads to high expression of CYP1A1. The highly active substances covalently bind to DNA, thereby inducing mutations and having a carcinogenic effect. CYP1A1 is highly expressed in a variety of tumor cells, not only directly promoting the proliferation and migration of cancer cells, but also increasing the drug resistance of cancer cells through its drug degradation characteristics, so it has been widely used as a tumor marker.

[0005] Inflammation is a very common and important basic pathological process in the body, and CYP1A1 is a hydroxylase mainly controlled by the inflammatory restricted aryl hydrocarbon receptor (AhR) and is involved in the metabolism of a broad spectrum of xenobiotics.

[0006] CYP1A1 negatively regulates the body's ability to resist sepsis caused by infection, inhibits the mobilization response of macrophages during infection, and is involved in the pathological processes of septic kidney injury and intestinal injury. Research has found that CYP1A1 can inhibit the "arginine-agmatine-polyamine / γ-aminobutyric acid" metabolic axis to inhibit the occurrence and development of sepsis, and thus can correct the disorder of the metabolic pathway by regulating the expression / activity of CYP1A1 to treat sepsis early and effectively.

[0007] In addition, CYP1A1 also plays an important role in oxidative stress injury and infections with Mycoplasma and Citrobacter rodentium.

[0008] CYP1A1 has been considered as a potential intervention target for the above various major diseases. Inhibiting CYP1A1 can effectively inhibit the occurrence and development of these diseases. The discovery of CYP1A1 inhibitors is particularly important for the design and development of its drugs.

[0009] The Global Burden of Bacterial Infections Report shows that in 2019, there were approximately 13.7 million deaths related to bacterial infections globally, with a mortality rate of 99.6 per 100,000 (74.2 - 132 per 100,000). These deaths related to bacterial infections accounted for 13.6% (10.1% - 18.1%) of the global deaths in 2019, making it the second leading cause of death globally, second only to ischemic heart disease.

[0010] The development of drugs against bacterial infections is generally considered difficult. Even if antibiotics are developed, drug-resistant bacteria are likely to emerge; when developing new antibiotics against these drug-resistant bacteria, new drug-resistant bacteria against the new antibiotics are likely to emerge, and so on. Eventually, the result may very likely be a situation where there are no drugs available for bacterial infections. Therefore, it is urgent to search for or develop new non-antibiotic anti-bacterial infection drugs. Summary of the Invention

[0011] On the one hand, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof,

[0012]

[0013] On the one hand, there is provided a method for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0014] On the one hand, there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a CYP1A1 inhibitor.

[0015] On the one hand, there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating or preventing tumors, bacterial infections, sepsis, inflammation, or organ damage.

[0016] The compound of formula (I) of the present application or a pharmaceutically acceptable salt thereof has been experimentally confirmed to be able to effectively inhibit CYP1A1 activity and is particularly effective against bacterial infections, sepsis, inflammation, or organ damage. Description of the Drawings

[0017] Figure 1 : (a) of Compound 1 1 1H NMR spectrum, (b) 13 13C NMR spectrum, and (c) 19 19F NMR spectrum.

[0018] Figure 2 : (a) of Compound 13 1 1H NMR spectrum and (b) 13 13C NMR spectrum.

[0019] Figure 3 : (a) of Compound 16 1 1H NMR spectrum and (b) 13 13C NMR spectrum.

[0020] Figure 4 : (a) of Compound 22 1 1H NMR spectrum and (b) 13 13C NMR spectrum.

[0021] Figure 5 : (a) of Compound 28 1 1H NMR spectrum and (b) 13 13C NMR spectrum.

[0022] Figure 6 : (a) of Compound 31 1 1H NMR spectrum and (b) 13 13C NMR spectrum.

[0023] Figure 7 : (a) of Compound 34 1 1H NMR spectrum and (b) 13 13C NMR spectrum.

[0024] Figure 8 : (a) of Compound 50 1 1H NMR spectrum and (b) 13 13C NMR spectrum.

[0025] Figure 9 : (a) of Compound 57 1 1H NMR spectrum and (b) 13 13C NMR spectrum.

[0026] Figure 10:(a) of Compound 58 1 H NMR spectrum and (b) 13 C NMR spectrum.

[0027] Figure 11 : Components of the compound of formula (I).

[0028] Figure 12 : EROD enzyme activities obtained by performing EROD assays on Caco-2 cells with (a) Compound 1, (b) Compound 6, (c) Compound 20, (d) Compound 25, (e) Compound 26, (f) Compound 29, (g) Compound 31, (h) Compound 34, (i) Compound 43, and (j) Compound 53 at various concentrations, and the IC 50 values calculated therefrom. The horizontal axis represents the concentration of each compound (μM), and the vertical axis represents the enzyme activity (optical density (OD)).

[0029] Figure 13 : EROD enzyme activity intensities obtained by performing EROD assays on CY h RAW cells with (a) Compound 1, (b) Compound 6, (c) Compound 20, (d) Compound 25, (e) Compound 26, (f) Compound 29, (g) Compound 31, (h) Compound 34, (i) Compound 43, and (j) Compound 53 at various concentrations, and the IC 50 values calculated therefrom. The horizontal axis represents the concentration of each compound (μM), and the vertical axis represents the enzyme activity (OD).

[0030] Figure 14 : EROD enzyme activity intensities obtained by performing EROD assays on (a, b) Compound 43, (c, d) Compound 31 at various concentrations using (a, c) Caco-2 cells or (b, d) CY h RAW cells. The horizontal axis, from left to right in sequence, is the negative control group ((without adding the compound), 50 nM group, 500 nM group, 5 μM group), and the vertical axis is the normalized relative enzyme activity based on the negative control group.

[0031] Figure 15 : Graph showing the relationship between survival time and survival rate of mice infected with (a) methicillin-resistant Staphylococcus aureus (MRSA), (b) Acinetobacter baumannii (AB), or (c) cecal ligation and puncture (CLP) treated with the compounds of the present application or bergapten. The horizontal axis represents the survival time (hours), and the vertical axis represents the survival rate (%). Solvent control group; Compound 43 group; Compound 31 group; Bergapten group.

[0032] Figure 16 : Mice infected with anti-(a, b) methicillin-resistant Staphylococcus aureus or (c, d) Acinetobacter baumannii were treated with the compounds of the present application, and the colony cultures of their peritoneal lavage fluids were taken. The obtained (a, c) photos and (b, d) colony count charts. In the (a, c) photos, the top row is the solvent control group, the middle row is the compound 43 group, and the bottom row is the compound 31 group; in the (b, d) colony count charts, the horizontal axis from left to right is the solvent control group, the compound 43 group, and the compound 31 group, and the vertical axis is the bacterial load (10 4 CFU).

[0033] Figure 17 : Mice infected with anti-methicillin-resistant Staphylococcus aureus were treated with the compounds of the present application, and the serum was taken for the measurement results of (a) aspartate aminotransferase (AST), (b) alanine aminotransferase (ALT), (c) creatinine (CRE), or (d) blood urea nitrogen (BUN). The horizontal axis from left to right is the blank control group, the solvent control group, the compound 43 group, and the compound 31 group, and the vertical axis is respectively (a) AST enzyme activity (Karman unit (karU)), (b) ALT enzyme activity (Karman unit (karU)), (c) creatinine concentration (μmol / L), (d) blood urea nitrogen concentration (mmol / L).

[0034] Figure 18 : Different concentrations of compound 43 were administered to mice, and the serum was taken for the measurement results of (a) aspartate aminotransferase (AST), (b) alanine aminotransferase (ALT), (c) creatinine, (d) blood urea nitrogen, (e) cardiac creatine kinase MB isoenzyme, (f) cardiac troponin I. The horizontal axis is the concentration of compound 43, and the vertical axis is respectively (a) AST enzyme activity (Karman unit (karU)), (b) ALT enzyme activity (Karman unit (karU)), (c) creatinine concentration (μmol / L), (d) blood urea nitrogen concentration (mmol / L), (e) CK-MB concentration (ng / mL), (f) cTn-I concentration (ng / L).

[0035] Figure 19 : After stimulating (a) Caco-2 cells and (b) CY h RAW cells with different concentrations of compound 43, the normalized cell viability values measured by the MTT assay, where the cell viability value of the group with a compound 43 concentration of 0 was set to 1. The horizontal axis is the concentration of compound 43, and the vertical axis is the normalized cell viability value. Detailed implementation mode

[0036] To further elaborate on the technical means and effects adopted by the present application to achieve the intended purpose, the following provides a detailed description of the specific implementation manners, structures, features, and effects thereof in accordance with the present application in conjunction with the accompanying drawings, various embodiments (including manufacturing examples and experimental examples), and preferred implementation manners.

[0037] Terms and Definitions

[0038] As used herein, the singular terms refer to one or more than one. For example, "element" or "an element" both refer to one element or more than one element. As used herein, the term "plurality" means at least two.

[0039] As used herein, all features or conditions defined in the form of numerical ranges or percentage ranges mentioned by endpoints are only for the sake of brevity and convenience. Therefore, the description of numerical ranges or percentage ranges should be regarded as having covered and specifically disclosed all possible sub-ranges and all individual integer and fractional values within the range, especially integer values. For example, the description of the range "1 to 8" should be regarded as having specifically disclosed all sub-ranges such as 1 to 7, 2 to 8, 2 to 6, 3 to 6, 4 to 8, 3 to 8, etc., especially the sub-ranges defined by all integer values, and should be regarded as having specifically disclosed the individual values such as 1, 2, 3, 4, 5, 6, 7, 8, etc. within the range; regardless of whether these ranges or these individual values are separately literally recited. Similarly, the description of the range "between 1 and 8" should also be regarded as having specifically disclosed all ranges such as 1 to 8, 1 to 7, 2 to 8, 2 to 6, 3 to 6, 4 to 8, 3 to 8, etc., and includes the endpoint values of these ranges, such as individual values 1, 2, 3, 4, 5, 6, 7, 8, etc.

[0040] As used herein, the term "about" means approximate, within the range of about or near. When the term "about" is used in combination with a numerical range, it modifies the range by expanding the boundaries above or below the provided value. Generally, the term "about" as used herein causes the numerical value to vary up and down by 10% from the provided value. On the one hand, the term "about" means plus or minus 20% of the numerical value of the number it modifies. For example, "about 50%" means within the range of 45% - 55%. It should also be understood that all integers and fractions are considered to be modified by the term "about". In this document, on the premise that the invention purpose can be achieved, the numerical value should be understood as having the accuracy of the effective digits of the said numerical value. For example, the number 40.0 can be understood as covering the range from 39.50 to 40.49.

[0041] As used herein, the terms "comprising", "including", or "containing", being non-exclusive or open-ended terms, are intended to mean that a combination (such as an apparatus, a composition, a method, etc.) includes the recited elements (such as each unit of the apparatus, each component of the composition, the substantial steps of the method, etc.), but does not exclude other elements. As used herein, the term "consisting essentially of", when used to define compositions and methods, means excluding other elements that have any substantial effect on the combination for the stated purpose, but not excluding other elements that do not substantially affect the basic and novel features of the present invention. As used herein, the term "consisting of" means excluding other combinations of elements (units, components, substantial steps, etc.), but unless otherwise stated, does not mean excluding trace amounts of unavoidable impurities. Embodiments defined by each of these connecting terms are within the scope of the present invention. As a specific embodiment thereof, a technical solution disclosed including the terms "comprising", "including", or "containing" should also be regarded as simultaneously disclosing the corresponding technical solutions including the terms "consisting essentially of" and "consisting of".

[0042] As used herein, the term "and / or" means and encompasses any and all possible combinations of one or more of the associated listed items. When used in a list of two or more items, the term "and / or" means that any one of the listed items can be included alone, or any combination of two or more of the listed items can be included. For example, if a group, combination, or composition, etc., is described as including (or containing) components A, B, C, and / or D, then the composition can include A alone, B alone, C alone, D alone, a combination of A and B, a combination of A and C, a combination of A and D, a combination of B and C, a combination of B and D, a combination of C and D, a combination of A, B, and C, a combination of A, B, and D, a combination of A, C, and D, a combination of B, C, and D, or a combination of A, B, C, and D.

[0043] As used herein, for any case where a Markush group or list of options is used to describe the features or embodiments of the present application, a subgroup or any individual member within the Markush group or list of options also constitutes a part of the present application respectively. For example, if it is recited herein that X "is selected from the group consisting of A, B, and C", it means that the technical solutions where X is A, X is B, X is C, X is A and / or B, X is A and / or C, X is B and / or C, and X is A and / or B and / or C are respectively disclosed where chemically permissible.

[0044] The compounds of the present application may contain asymmetric or chiral centers and thus exist in different stereoisomeric forms. All stereoisomers of the compounds of the present application, including but not limited to diastereoisomers, enantiomers, and atropisomers, and mixtures thereof such as racemic mixtures, are considered to form part of the present application.

[0045] As used herein, the term "(substituted with a substituent)" means that one or more hydrogens on the specified group are replaced by a specific group, radical, or moiety, i.e., a substituent, provided that such replacement is chemically permissible, such that there is one or more of the said substituents on the specified group. As used herein, the term "substituent" refers to a chemical group, radical, or moiety that is covalently attached or (where appropriate) fused to a parent group or moiety. As used herein, the term "optionally (substituted with a substituent)" means that one or more hydrogens on the specified group are optionally replaced by a substituent, which includes the alternative technical solutions of "substituted with a substituent" and "unsubstituted", and is equivalent to the term "substituted or unsubstituted with a substituent". This term can further define the substituent as a specified substituent. For example, the term "optionally methyl-substituted" means optionally substituted with a methyl group, which includes the alternative technical solutions of "substituted with a methyl group" and "unsubstituted", and is equivalent to the term "substituted or unsubstituted with a methyl group". This term can also be further modified by a quantifier. For example, the term "optionally mono-substituted" means optionally substituted with one substituent, which includes the alternative technical solutions of "substituted with one substituent" and "unsubstituted", and is equivalent to the term "substituted or unsubstituted with one substituent"; the term "optionally methyl mono-substituted" means optionally substituted with one methyl group, which includes the alternative technical solutions of "substituted with one methyl group" and "unsubstituted", and is equivalent to the term "substituted or unsubstituted with one methyl group".

[0046] As used herein, the term "pharmaceutically acceptable salt" may refer to a salt prepared from a pharmaceutically acceptable non-toxic base or acid.

[0047] When the compound is acidic, its corresponding salts can be prepared from pharmaceutically acceptable non-toxic bases (including inorganic and organic bases). Salts derived from inorganic bases include, but are not limited to, aluminum salts, ammonium salts, calcium salts, copper salts, cuprous salts, iron salts, ferrous salts, lithium salts, magnesium salts, potassium salts, sodium salts, zinc salts, etc., especially ammonium salts, calcium salts, magnesium salts, potassium salts, and sodium salts. Salts derived from organic bases include salts of primary, secondary, and tertiary amines, as well as cyclic amines and substituted amines (such as naturally occurring and synthetic substituted amines). Salts derived from organic bases include, but are not limited to, salts of arginine, betaine, caffeine, choline, N′,N′-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucosamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc.

[0048] When the compound is basic, its corresponding salts can be prepared from pharmaceutically acceptable non-toxic acids (including inorganic and organic acids). Salts derived from organic acids include, but are not limited to, acetate, benzenesulfonate, benzoate, camphorsulfonate, citrate, ethanesulfonate, fumarate, gluconate, glutamate, hydrobromide, hydrochloride, isethionate, lactate, maleate, malate, mandelate, mesylate, mucate, nitrate, pamoate, pantothenate, phosphate, succinate, sulfate, tartrate, p-toluenesulfonate, etc.

[0049] As used herein, the term "inhibit" means to prevent, reduce, suppress, or inactivate the physiological function (i.e., activity) of one or more specific proteins, such as CYP1A1, either completely or in part; the term "inhibitor" means a substance (e.g., a small molecule compound, but not limited thereto) that can prevent, reduce, suppress, or inactivate the physiological function (i.e., activity) of one or more specific proteins, such as CYP1A1, either completely or in part. For example, a "CYP1A1 inhibitor" is a substance that prevents, reduces, suppresses, or inactivates the physiological function (i.e., activity) of CYP1A1. A CYP1A1 inhibitor can inhibit the activity of CYP1A1 by binding to or interacting with CYP1A1, but not limited thereto. A CYP1A1 inhibitor can also prevent or reduce the activity of CYP1A1 by preventing or reducing the expression of the CYP1A1 gene. A CYP1A1 inhibitor can cause a decrease in the expression level of CYP1A1, such as a decrease in the level of CYP1A1 mRNA or CYP1A1 protein, further resulting in a decrease in the activity of CYP1A1. Inhibitors of CYP1A1 can include transcriptional repressors of CYP1A1 expression that can reduce the level of CYP1A1. All means and methods that result in a decrease in the activity of CYP1A1 (possibly due to lower expression) should be considered part of the inhibitors of CYP1A1 as used herein.

[0050] As used herein, the term "treat" can refer to eradicating, inhibiting, alleviating, or delaying a disease, disorder, or condition, such as eradicating, inhibiting, alleviating, or delaying the development of at least one of its clinical symptoms, or alleviating or improving at least one physical parameter, including those that may not be recognized by the patient. "Treat" can be to modulate the disease, disorder, or condition physically (e.g., stabilization of discernible symptoms), physiologically (e.g., stabilization of a physical parameter), or both. "Treat" can be to slow the onset or rate of development of a condition, disorder, or disease, reduce or alleviate the symptoms associated therewith, produce a complete or partial regression of the condition, or a combination thereof. The results obtained by "treatment" include, but are not limited to, reduction or alleviation of symptoms, diminution of the extent of the disease, stabilization of the disease state (i.e., not worsening), delay or slowing of disease progression, improvement or alleviation of the disease state, and remission (partial or complete), whether detectable or not. "Treat" can also refer to an extended survival relative to the expected survival time without treatment. "Treat" can also include an increased inhibition or reduction in the severity of a pathological state or symptoms relative to the absence of treatment, and does not necessarily imply complete cessation of the disease, disorder, or condition. An individual (e.g., a human) in need of treatment can be an individual already suffering from the relevant disease, disorder, or condition.

[0051] As used herein, the term "prevention" can refer to eradicating, inhibiting, alleviating, or delaying the development of the risk of a disease, disorder, or condition, or eradicating, inhibiting, alleviating, or improving the underlying disease, disorder, or condition being prevented. The individual (e.g., a human) in need of prevention can be an individual who has not yet developed the relevant disease, disorder, or condition. "Prevention" includes delaying the onset of a disease relative to when treatment has not been administered to the individual, and does not necessarily mean that the relevant disease, disorder, or condition is permanently prevented. To obtain the benefits of prevention, a pharmaceutical composition can be administered to a patient at risk of developing a specific disease or to a patient having one or more physical symptoms of a disease, even if the disease has not been diagnosed. Unless otherwise specified, methods for evaluating the therapeutic and / or preventive effects of a disease are generally known in the art.

[0052] Embodiments

[0053] Compound

[0054] In some embodiments, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0055]

[0056] R 1 may be an optionally substituted aryl, heteroaryl, arylvinyl, arylethynyl, trimethylsilylethynyl, or cycloalkyl.

[0057] Said optional substitution may mean substitution by a substituent, i.e., R 1 may be an aryl, heteroaryl, arylvinyl, arylethynyl, trimethylsilylethynyl, or cycloalkyl having one or more substituents thereon.

[0058] In some embodiments, the substituent may be: a halogen (such as fluorine, chlorine, bromine, iodine); an alkyl group (such as C 1-6 alkyl, especially C 1-4 alkyl, more particularly methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, etc.); a cycloalkyl group (such as C 3-8 cycloalkyl, especially cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, etc.); an alkenyl group (such as C 2-6 alkenyl, especially vinyl, prop-1-enyl, prop-2-enyl, but-1-enyl, but-2-enyl, but-3-enyl, etc.); an alkynyl group (such as C 2-6Alkenyl, especially ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl, but-3-ynyl, etc.); aryl (such as phenyl, naphthyl, anthryl, etc.); aralkyl (such as benzyl, phenethyl, etc.); amino; primary amino (such as methylamino, ethylamino, propylamino, butylamino, phenylamino, etc.); secondary amino (such as dimethylamino, diethylamino, diphenylamino, etc.); aminoalkyl (such as aminomethyl, aminoethyl, aminopropyl, aminobutyl, etc.); acylamino (such as formylamino, acetylamino, propionylamino, butyrylamino, benzoylamino, etc.); cyano; nitro; nitroso; hydroxy; alkoxy (such as methoxy, ethoxy, propoxy, butoxy, phenoxy, etc.); trifluoromethoxy; hydroxyalkyl (such as hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, etc.); acyl (such as formyl, acetyl, propionyl, butyryl, benzoyl, etc.); carbamoyl; carboxy; alkoxycarbonyl (such as tert-butoxycarbonyl); aryloxycarbonyl (such as benzyloxycarbonyl); phosphino; primary phosphino (such as methylphosphino, ethylphosphino, propylphosphino, butylphosphino, phenylphosphino, etc.); secondary phosphino (such as dimethylphosphino, diethylphosphino, diphenylphosphino, etc.); phosphinoalkyl (such as phosphinomethyl, phosphinoethyl, phosphinopropyl, phosphinobutyl, etc.); phosphoryl; phosphonous; mercapto; alkylthio (such as methylthio, ethylthio, propylthio, butylthio, phenylthio, etc.); mercaptoalkyl (such as mercaptomethyl, mercaptoethyl, mercaptopropyl, mercaptobutyl, etc.); sulfonyl (such as methylsulfonyl, ethylsulfonyl, propylsulfonyl, butylsulfonyl, benzylsulfonyl, etc.); sulfinyl (such as methylsulfinyl, ethylsulfinyl, propylsulfinyl, butylsulfinyl, benzylsulfinyl, etc.); sulfonic acid group; sulfinic acid group; boranyl; boronic acid group; optionally substituted silyl (for example, trimethylsilyl); or optionally substituted siloxy (for example, tert-butyldimethylsiloxy), but not limited thereto. In some embodiments, the substituent is further substituted with one or more additional substituents.

[0059] In some embodiments, R 1 may be an optionally substituted aryl, especially an optionally substituted phenyl, an optionally substituted naphthyl, or an optionally substituted anthryl. In some embodiments, the optionally substituted aryl (such as an optionally substituted phenyl, naphthyl, or anthryl) may be a halogen- and / or alkynyl-substituted aryl (such as a halogen- and / or alkynyl-substituted phenyl, naphthyl, or anthryl), especially a halogen-substituted aryl (such as a halogen-substituted phenyl, naphthyl, or anthryl), or especially an alkynyl-substituted aryl (such as an alkynyl-substituted phenyl, naphthyl, or anthryl). In some embodiments, the optionally substituted phenyl (including the optionally substituted phenyl in optionally substituted styryl) may be a halogen- and / or alkynyl-substituted phenyl, especially a halogen-substituted phenyl, or especially an alkynyl-substituted phenyl. In some embodiments, the optionally substituted phenyl may be Wherein: n is 0, 1, 2, 3, 4, or 5, especially 1, 2, 3, 4, or 5, more especially 1; each X is independently selected from the substituents described above, especially halogen or alkynyl (such as fluorine or alkynyl), more especially halogen (such as fluorine), or especially alkynyl.

[0060] In some embodiments, R 1 or an optionally substituted phenyl group may be a fluorophenyl group (such as 2-fluorophenyl, 3-fluorophenyl, or 4-fluorophenyl), a difluorophenyl group (such as 2,3-difluorophenyl, 2,4-difluorophenyl, 2,5-difluorophenyl, 2,6-difluorophenyl, 3,4-difluorophenyl, or 3,5-difluorophenyl), a trifluorophenyl group (such as 2,3,4-trifluorophenyl, 2,3,5-trifluorophenyl, 2,3,6-trifluorophenyl, 2,4,5-trifluorophenyl, 2,4,6-trifluorophenyl, or 3,4,5-trifluorophenyl), a tetrafluorophenyl group (such as 2,3,4,5-tetrafluorophenyl, 2,3,4,6-tetrafluorophenyl, or 2,3,5,6-tetrafluorophenyl), or a pentafluorophenyl group (2,3,4,5,6-pentafluorophenyl).

[0061] In some embodiments, R 1 or an optionally substituted phenyl group may be an ethynylphenyl group (such as 2-ethynylphenyl, 3-ethynylphenyl, or 4-ethynylphenyl), a diethynylphenyl group (such as 2,3-diethynylphenyl, 2,4-diethynylphenyl, 2,5-diethynylphenyl, 2,6-diethynylphenyl, 3,4-diethynylphenyl, or 3,5-diethynylphenyl), a triethynylphenyl group (such as 2,3,4-triethynylphenyl, 2,3,5-triethynylphenyl, 2,3,6-triethynylphenyl, 2,4,5-triethynylphenyl, 2,4,6-triethynylphenyl, or 3,4,5-triethynylphenyl), a tetraethynylphenyl group (such as 2,3,4,5-tetraethynylphenyl, 2,3,4,6-tetraethynylphenyl, or 2,3,5,6-tetraethynylphenyl), or a pentaethynylphenyl group (2,3,4,5,6-pentaethynylphenyl).

[0062] In some embodiments, the optionally substituted phenyl group may have a boronic acid group substituent. In some embodiments, R 1 may be a boronic acid group fluorophenyl, for example, 4-boronic acid group-2-fluorophenyl.

[0063] In some embodiments, R 1 may be an optionally substituted heteroaryl group. In some embodiments, R 1Or the optionally substituted heteroaryl may be optionally substituted, especially optionally methyl-substituted, azo, oxo, thio, azoxo, or azothio, five-membered heterocyclic group, six-membered heterocyclic group, six-membered ring-fused five-membered heterocyclic group (such as benzo five-membered heterocyclic group), or six-membered ring-fused six-membered heterocyclic group (such as benzo six-membered heterocyclic group). In some embodiments, R 1 Or the optionally substituted heteroaryl may be optionally methyl-substituted, azo, oxo, or thio, five-membered heterocyclic group, six-membered heterocyclic group, benzo five-membered heterocyclic group, or benzo six-membered heterocyclic group.

[0064] In some embodiments, R 1 may be an optionally substituted pyrrolyl, furyl, thienyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyranyl, thiopyranyl, pyridazinyl, pyrimidinyl, pyrazinyl, oxazinyl, thiazinyl, benzopyrrolyl (such as indolyl or isoindolyl), benzofuryl, benzothienyl, benzopyrazolyl, benzimidazolyl (such as indazolyl), benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzopyridyl (such as quinolinyl or isoquinolinyl), benzopyranyl (such as chromenyl or isochromenyl), benzothiopyranyl, benzopyridazinyl (such as phthalazinyl or cinnolinyl), benzopyrimidinyl (quinazolinyl), benzopyrazinyl (quinoxalinyl), benzoxazinyl, benzothiazinyl, purinyl, naphthyridinyl, or pteridinyl. In some embodiments, R 1 may be an optionally substituted pyridyl, pyrrolyl, quinolinyl, indolyl, furyl, benzofuryl, thienyl, or benzothienyl.

[0065] In some embodiments, R 1 may be an optionally substituted pyridyl (such as optionally substituted pyridin-2-yl, pyridin-3-yl, or pyridin-4-yl), pyrrolyl (such as optionally substituted 1H-pyrrol-2-yl or 1H-pyrrol-3-yl), quinolinyl (such as optionally substituted quinolin-2-yl, quinolin-3-yl, or quinolin-4-yl), indolyl (such as optionally substituted 1H-indol-2-yl or 1H-indol-3-yl), or thienyl (such as optionally substituted thiophen-2-yl or thiophen-3-yl), especially an optionally methyl-monosubstituted pyridyl, pyrrolyl, quinolinyl, indolyl, or thienyl. In some embodiments, R 1 may be an optionally substituted pyridyl, especially methylpyridyl, more especially 6-methylpyridin-3-yl. In some embodiments, R 1 may be an optionally substituted pyrrolyl, especially methylpyrrolyl, more especially 1-methyl-1H-pyrrol-2-yl. In some embodiments, R 1 may be an optionally substituted quinolinyl, especially quinolinyl, more especially quinolin-2-yl or quinolin-3-yl. In some embodiments, R 1may be an optionally substituted indolyl group, especially a methylindolyl group, more especially a 1-methyl-1H-indol-3-yl group. In some embodiments, R 1 may be an optionally substituted thienyl group, especially a thienyl group, more especially a thien-2-yl or thien-3-yl group.

[0066] In some embodiments, R 1 may be R 1 may be: a phenyl, naphthyl, or anthracenyl group substituted with halogen and / or alkynyl; or an optionally methyl-substituted, azo, oxa, or thia, five-membered heterocyclic group, six-membered heterocyclic group, benzo five-membered heterocyclic group, or benzo six-membered heterocyclic group.

[0067] In some embodiments, R 1 may be an optionally substituted arylvinyl group, especially an optionally substituted styryl group, an optionally substituted naphthylvinyl group, or an optionally substituted anthracenylvinyl group. In some embodiments, R 1 or the optionally substituted arylvinyl group may be: –C═C–Q, where Q is an optionally substituted aryl group, especially an optionally substituted phenyl group, and may be an optionally substituted aryl group or an optionally substituted phenyl group as defined above. In some embodiments, R 1 or the optionally substituted arylvinyl group may be an optionally substituted (E)-arylviny or an optionally substituted (Z)-arylviny especially an optionally substituted (E)-arylviny

[0068] In some embodiments, R 1 or the optionally substituted styryl group may be an optionally substituted (E)-styryl group, especially an (E)-styryl group substituted with halogen and / or alkynyl, more especially an (E)-styryl group substituted with halogen. In some embodiments, the optionally substituted styryl group may be fluorostyryl (such as 2-fluorostyryl, 3-fluorostyryl, or 4-fluorostyryl), difluorostyryl (such as 2,3-difluorostyryl, 2,4-difluorostyryl, 2,5-difluorostyryl, 2,6-difluorostyryl, 3,4-difluorostyryl, or 3,5-difluorostyryl), trifluorostyryl (such as 2,3,4-trifluorostyryl, 2,3,5-trifluorostyryl, 2,3,6-trifluorostyryl, 2,4,5-trifluorostyryl, 2,4,6-trifluorostyryl, or 3,4,5-trifluorostyryl), tetrafluorostyryl (such as 2,3,4,5-tetrafluorostyryl, 2,3,4,6-tetrafluorostyryl, or 2,3,5,6-tetrafluorostyryl), or pentafluorostyryl (2,3,4,5,6-pentafluorostyryl).

[0069] In some embodiments, R 1May be an optionally substituted arylethynyl, particularly an optionally substituted phenylethynyl, an optionally substituted naphthylethynyl, or an optionally substituted anthrylethynyl. In some embodiments, R 1 Or the optionally substituted arylethynyl may be: –C≡C–Q, where Q is an optionally substituted aryl, particularly an optionally substituted phenyl, and may be an optionally substituted aryl or an optionally substituted phenyl as defined above.

[0070] In some embodiments, R 1 Or the optionally substituted phenylethynyl may be a halogen- and / or alkynyl-substituted phenylethynyl, particularly a halogen-substituted phenylethynyl. In some embodiments, the optionally substituted phenylethynyl may be fluorophenylethynyl (such as 2-fluorophenylethynyl, 3-fluorophenylethynyl, or 4-fluorophenylethynyl), difluorophenylethynyl (such as 2,3-difluorophenylethynyl, 2,4-difluorophenylethynyl, 2,5-difluorophenylethynyl, 2,6-difluorophenylethynyl, 3,4-difluorophenylethynyl, or 3,5-difluorophenylethynyl), trifluorophenylethynyl (such as 2,3,4-trifluorophenylethynyl, 2,3,5-trifluorophenylethynyl, 2,3,6-trifluorophenylethynyl, 2,4,5-trifluorophenylethynyl, 2,4,6-trifluorophenylethynyl, or 3,4,5-trifluorophenylethynyl), tetrafluorophenylethynyl (such as 2,3,4,5-tetrafluorophenylethynyl, 2,3,4,6-tetrafluorophenylethynyl, or 2,3,5,6-tetrafluorophenylethynyl), or pentafluorophenylethynyl (2,3,4,5,6-pentafluorophenylethynyl).

[0071] In some embodiments, R 1 May be trimethylsilylethynyl.

[0072] In some embodiments, R 1 Cycloalkyl. In some embodiments, R 1 May be C 3-8 Cycloalkyl, particularly cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl.

[0073] In some embodiments, R 1may be phenyl, 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2,4-difluorophenyl, 3,5-difluorophenyl, 2,4,5-trifluorophenyl, 2,3,4,5,6-pentafluorophenyl, 4-borono-2-fluorophenyl, 2-chloro-4-fluorophenyl, 2-ethynylphenyl, 3-ethynylphenyl, 4-ethynylphenyl, 4-tert-butylphenyl, 4-biphenylyl, 4-cyanophenyl, 4-(dimethylamino)phenyl, 4-nitrophenyl, 4-butoxyphenyl, 3-(trifluoromethoxy)phenyl, 4-(tert-butyldimethylsilyloxy)phenyl, 2-(diphenylphosphino)phenyl, 4-methylthiophenyl, 1-naphthyl, 9-anthryl, 1-(tert-butoxycarbonyl)-1H-pyrrol-2-yl, 1-methyl-1H-pyrrol-2-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, 6-methylpyridin-3-yl, 2-bromopyridin-3-yl, pyrimidin-2-yl, quinolin-2-yl, quinolin-3-yl, 1-acetyl-1H-indol-3-yl, 1-methyl-1H-indol-3-yl, 1-benzyl-1H-indol-3-yl, 1-benzyl-5-methoxy-1H-indol-3-yl, furan-3-yl, furan-2-yl, benzo[b]furan-3-yl, benzo[b]furan-2-yl, 5-nitrofuran-2-yl, 5-iodofuran-2-yl, thiophen-2-yl, thiophen-3-yl, benzo[b]thiophen-2-yl, cyclopropyl, (E)-styryl, (E)-4-fluorostyryl, phenylacetylene, or trimethylsilylethynyl.

[0074] In some embodiments, R 1 may be fluorophenyl, trifluorophenyl, or ethynylphenyl, or a pyridinyl, pyrrolyl, quinolinyl, indolyl, or thienyl optionally monosubstituted with methyl; particularly fluorophenyl or ethynylphenyl.

[0075] In some embodiments, R 1may be 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2,4,5-trifluorophenyl, 2-ethynylphenyl, 3-ethynylphenyl, 4-ethynylphenyl, 6-methylpyridin-3-yl, 1-methyl-1H-pyrrol-2-yl, 1-methyl-1H-indol-3-yl, quinolin-2-yl, quinolin-3-yl, thiophen-2-yl, or thiophen-3-yl; particularly 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2,4,5-trifluorophenyl, 2-ethynylphenyl, 3-ethynylphenyl, 4-ethynylphenyl, 6-methylpyridin-3-yl, 1-methyl-1H-pyrrol-2-yl, 1-methyl-1H-indol-3-yl, quinolin-2-yl, or thiophen-3-yl; more particularly 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2,4,5-trifluorophenyl, 2-ethynylphenyl, 3-ethynylphenyl, 4-ethynylphenyl, 6-methylpyridin-3-yl, 1-methyl-1H-pyrrol-2-yl, or 1-methyl-1H-indol-3-yl; even more particularly 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2-ethynylphenyl, 3-ethynylphenyl, or 4-ethynylphenyl.

[0076] In some embodiments, R 1 may be 2-ethynylphenyl or 1-methyl-1H-indol-3-yl.

[0077] Preparation method

[0078] In one aspect, there is provided a method for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0079] The method for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof may include obtaining the compound of formula (I) or a pharmaceutically acceptable salt thereof from a compound of formula (Ic) (5-(2-(triphenyl-λ 5 -phosphoranylidene)acetyl)-1,3-dimethyl-1,3-dihydro-2H-benzo[d]imidazol-2-one), particularly reacting the compound of formula (Ic) with R 1 –CHO to obtain the compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, obtaining the compound of formula (I) may include mixing the compound of formula (Ic) and R 1 –CHO in a solvent. In some embodiments, the solvent may be toluene. In some embodiments, the equivalent ratio of the compound of formula (Ic) to R 1 –CHO may be 1:(0.8–1.2), particularly 1:1.

[0080] The method for preparing the compound of formula (I) or a pharmaceutically acceptable salt thereof may further include obtaining the compound of formula (Ic) from the compound of formula (Ib) (5-(2-chloroacetyl)-1,3-dimethyl-1,3-dihydro-2H-benzo[d]imidazol-2-one), particularly reacting the compound of formula (Ib) with triphenylphosphine to obtain the compound of formula (Ic). In some embodiments, obtaining the compound of formula (Ic) may include mixing the compound of formula (Ia) and triphenylphosphine in a solvent. In some embodiments, the solvent may be acetonitrile. In some embodiments, the equivalent ratio of the compound of formula (Ia), aluminum trichloride, and chloroacetyl chloride may be 1:(1.0–1.2), particularly 1:1.1.

[0081] The method for preparing the compound of formula (I) or a pharmaceutically acceptable salt thereof may further include obtaining the compound of formula (Ib) from the compound of formula (Ia) (1,3-dimethyl-1,3-dihydro-2H-benzo[d]imidazol-2-one), particularly reacting the compound of formula (Ia) with chloroacetyl chloride to obtain the compound of formula (Ib). In some embodiments, obtaining the compound of formula (Ib) may include mixing the compound of formula (Ia), aluminum trichloride, and chloroacetyl chloride in a solvent. In some embodiments, the solvent may be dichloromethane. In some embodiments, the equivalent ratio of the compound of formula (Ia), aluminum trichloride, and chloroacetyl chloride may be 1:(1.0–1.5):(1.0–1.2), particularly 1:1.2:1.08.

[0082]

[0083] Use

[0084] In one aspect, there is provided the use of the compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a CYP1A1 inhibitor. In one aspect, there is provided the use of the compound of formula (I) or a pharmaceutically acceptable salt thereof in inhibiting CYP1A1. In one aspect, there is provided a pharmaceutical composition (CYP1A1 inhibitor) for inhibiting CYP1A1, comprising the compound of formula (I) or a pharmaceutically acceptable salt thereof. In one aspect, there is provided the compound of formula (I) or a pharmaceutically acceptable salt thereof for inhibiting CYP1A1. In one aspect, there is provided the compound of formula (I) or a pharmaceutically acceptable salt thereof when inhibiting CYP1A1. In one aspect, there is provided a method for inhibiting CYP1A1, comprising administering the compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0085] In one aspect, there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or preventing a disease or disorder. In one aspect, there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in treating or preventing a disease or disorder. In one aspect, there is provided a pharmaceutical composition for treating or preventing a disease or disorder, comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof. In one aspect, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for treating or preventing a disease or disorder. In one aspect, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof in treating or preventing a disease or disorder. In one aspect, there is provided a method of treating or preventing a disease or disorder, comprising administering a compound of formula (I) or a pharmaceutically acceptable salt thereof. The disease or disorder may be a tumor, a bacterial infection, sepsis, an inflammation, or an organ injury, particularly a tumor, or particularly a bacterial infection or sepsis, inflammation, or organ injury caused thereby.

[0086] In some embodiments, the disease or disorder is a tumor. In some embodiments, the tumor is a cancer or a pre-cancerous adenoma. In some embodiments, the tumor is a solid tumor. In some embodiments, the tumor is a hematological tumor. In some embodiments, the tumor (or the cancer) is liver cancer (such as hepatocellular carcinoma or cholangiocarcinoma), gastric cancer (such as gastric adenocarcinoma, gastric lymphoma, gastric leiomyosarcoma, gastrointestinal stromal tumor, or neuroendocrine cell carcinoid), ovarian cancer, endometrial cancer, cervical cancer, colorectal cancer (such as cecal cancer, appendiceal cancer, ascending colon cancer, transverse colon cancer, descending colon cancer, sigmoid colon cancer, rectal cancer, or anal cancer), lung cancer (such as squamous cell lung cancer, non-small cell lung cancer, or small cell lung cancer), soft tissue sarcoma, osteosarcoma, fibrosarcoma, skin cancer (such as malignant melanoma), testicular cancer, breast cancer, fibrosarcoma, neuroblastoma, brain cancer (such as glioma, such as ependymoma, astrocytoma, oligodendroglioma, brainstem glioma, or oligoastrocytoma), bladder cancer, bowel cancer, prostate cancer, kidney cancer (such as renal cell carcinoma or renal pelvic carcinoma), pancreatic cancer (such as pancreatic adenocarcinoma, pancreatic acinar cell carcinoma, pancreatic cystadenocarcinoma, pancreatoblastoma, pancreatic squamous cell carcinoma, pancreatic signet ring cell carcinoma, pancreatic hepatoid cell carcinoma, pancreatic colloid carcinoma, or undifferentiated pancreatic carcinoma), pleural mesothelioma, head and neck squamous cell carcinoma, nasopharyngeal carcinoma, oropharyngeal carcinoma, or blood cancer (such as acute myeloid leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, or lymphoma).

[0087] In some embodiments, the disease or disorder is a bacterial infection. In some embodiments, the disease or disorder is sepsis, particularly sepsis caused by a bacterial infection. In some embodiments, the disease or disorder is inflammation, particularly inflammation caused by a bacterial infection. In some embodiments, the disease or disorder is organ damage, particularly organ damage caused by a bacterial infection or sepsis resulting therefrom. In some embodiments, the organ damage includes oxidative stress damage.

[0088] In some embodiments, the bacteria may be: Gram-positive bacteria, including but not limited to Staphylococcus aureus (such as methicillin-resistant Staphylococcus aureus), Streptococcus pneumoniae, Enterococcus faecalis, Enterococcus faecium, Enterococcus avium, or Mycobacterium tuberculosis; or Gram-negative bacteria, including but not limited to Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Mycobacterium tuberculosis, Morganella morganii, Providencia stuartii, or Pseudomonas aeruginosa. In some embodiments, the bacteria are Staphylococcus aureus (particularly methicillin-resistant Staphylococcus aureus) or Acinetobacter baumannii.

[0089] In some embodiments, the inflammation and / or organ damage is inflammation or damage of one or more of the skin, muscle, bone, brain, cerebellum, brainstem, spinal cord, eye, ear, nose, tongue, heart, blood vessels, throat, trachea, lung, esophagus, stomach, small intestine, large intestine, liver, gallbladder, pancreas, kidney, bladder, urethra, ovary, uterus, testis, spleen, thymus, lymphatic vessels, or lymph nodes, particularly one or more of the liver, kidney, or heart, but not limited thereto.

[0090] Examples

[0091] Unless otherwise specified, the reagents used in the embodiments of the present application (including each manufacturing example and experimental example) are all of analytical grade, and the progress of all chemical reactions is detected by thin-layer chromatography (TLC).

[0092] Preparation of the Compound of Formula (I) in Production Example 1

[0093] In some embodiments, the preparation method of the compound of formula (I) is as shown in formula (i).

[0094]

[0095] The preparation method of the compound of formula (I) may include:

[0096] - Preparing the compound of formula (Ib) (5-(2-chloroacetyl)-1,3-dimethyl-1,3-dihydro-2H-benzo[d]imidazol-2-one) from the compound of formula (Ia) (1,3-dimethyl-1,3-dihydro-2H-benzo[d]imidazol-2-one);

[0097] - Preparation of the compound of formula (Ic) (5-(2-(triphenyl-λ 5 - phosphinylidene)acetyl)-1,3-dimethyl-1,3-dihydro-2H-benzo[d]imidazol-2-one) from the compound of formula (Ib); and

[0098] - Preparation of the compound of formula (I) from the compound of formula (Ic).

[0099]

[0100] 1.1 Preparation of the compound of formula (Ib) from the compound of formula (Ia)

[0101] Weigh the compound of formula (Ia) and place it in a flask. Add dichloromethane, and then add anhydrous aluminum trichloride. Stir at 30 °C for 30 minutes. Cool it to 0 °C in an ice bath, and while stirring, dropwise add chloroacetyl chloride. Stir for another 30 minutes, and then stir at room temperature for 12 hours. Among them, the equivalent ratio of the compound of formula (Ia), aluminum trichloride, and chloroacetyl chloride is 1:1.2:1.08.

[0102] Monitor the reaction progress by TLC. After the reaction is complete, quench the reaction with 1 N / mol hydrochloric acid, extract with dichloromethane at 0 °C - 5 °C. Wash the obtained organic phase with saturated sodium bicarbonate solution, and then wash with ice water and brine respectively. Combine the organic layers, remove water, rotary evaporate to dryness, and separate by column chromatography to obtain the compound of formula (Ib).

[0103] 1.2 Preparation of the compound of formula (Ic) from the compound of formula (Ib)

[0104] Weigh the compound of formula (Ib) and triphenylphosphine and place them in a three-necked flask. Under argon protection, add dry acetonitrile solvent to it with a syringe. Place the reaction flask in an oil bath at 83 °C and react overnight. Among them, the equivalent ratio of the compound of formula (Ib) and triphenylphosphine is 1:1.1.

[0105] Monitor the reaction progress by TLC. After the reaction is complete, cool it to room temperature, quench the reaction with saturated sodium carbonate solution, extract with ethyl acetate, remove water, rotary evaporate to dryness, and separate by column chromatography to obtain the compound of formula (Ic).

[0106] 1.3 Preparation of the compound of formula (I) from the compound of formula (Ic)

[0107] Dissolve the compound of formula (Ic) (0.1 mmol, 1.0 eq) and R 1 –CHO (0.1 mmol, 1.0 eq) in 1 mL of toluene, and reflux at 120 °C overnight. Remove the solvent by rotary evaporation and purify by column chromatography to obtain the compound of formula (I).

[0108] The various compounds of formula (I) prepared in this Preparation Example are shown as the following Compound 1 to Compound 53.

[0109]

[0110]

[0111] The preparation methods of some representative compounds of formula (I) in this application are exemplified below. Those skilled in the art should be able to analogously know the preparation methods of other compounds of formula (I) according to the specific preparation methods of the following compounds, combined with the conventional preparation and purification means in the art.

[0112] (E)-5-(3-(4-Fluorophenyl)acryloyl)-1,3-dimethyl-1,3-dihydro-2H-benzo[d]imidazol-2-one Preparation of (1)

[0113]

[0114] Dissolve the compound of formula (Ic) (0.1 mmol, 1.0 eq) and p-fluorobenzaldehyde (0.1 mmol, 1.0 eq) in 1 mL of toluene, and reflux the reaction overnight at 120 °C. Rotate evaporate to remove the solvent and concentrate the mixture, and separate the target product by column chromatography (dichloromethane∶methanol = 300∶1) to obtain Compound 1. The yield is 72.2%, a pale yellow solid.

[0115] Use 1 1H NMR, 13 13C NMR, and 19 19F NMR to determine the structure of Compound 1 and detect the molecular weight by HRMS-EI. The NMR spectra are as Figure 1 , and the data are as follows.

[0116] 1 1H NMR (600 MHz, CDCl 3 3) δ 7.86 (dt, J = 8.3, 1.4 Hz, 1H), 7.80 (d, J = 15.6 Hz, 1H), 7.71 (d, J = 1.4 Hz, 1H), 7.65 (dd, J = 8.4, 5.4 Hz, 2H), 7.52 (d, J = 15.7 Hz, 1H), 7.11 (t, J = 8.4 Hz, 2H), 7.03 (d, J = 8.2 Hz, 1H), 3.47 (dd, J = 10.8, 1.1 Hz, 6H).

[0117] 13 13C NMR (150 MHz, CDCl 3)δ 188.70, 163.97 (d, J = 251.8 Hz), 154.83, 142.91, 134.06, 131.89, 130.25 (d, J = 8.4 Hz), 123.31, 121.48 (d, J = 2.4 Hz), 116.07 (d, J = 22.0 Hz), 107.41, 106.48, 27.39, 27.35.

[0118] 19 F NMR (564 MHz, CDCl 3 ) δ -109.20.

[0119] HRMS-EI (m / z): C 18 H 15 FN 2 O 2 of [M + H] + Calculated: 311.1190; Found: 311.1188.

[0120] ( E )-5-(3-(2-Naphthyl)acryloyl)-1,3-dimethyl-1,3-dihydro-2 H -benzo d imidazol-2-one Preparation of (13)

[0121]

[0122] The compound of formula (Ic) (0.1 mmol, 1.0 eq), 2-naphthaldehyde (0.1 mmol, 1.0 eq) were dissolved in 1 mL of toluene, and the reaction was refluxed at 120 °C overnight. The solvent was removed by rotary evaporation and the mixture was concentrated. The target product, namely compound 13, was separated by column chromatography (dichloromethane∶methanol = 200∶1). The yield was 52%, a pale yellow solid.

[0123] Using 1 H NMR and 13 C NMR to determine the structure of compound 13 and the molecular weight was detected by HRMS-EI. The NMR spectra are as Figure 2 , and the data are as follows.

[0124] 1 H NMR (600 MHz, CDCl 3δ 8.06–7.99 (m, 2H), 7.92 (dd, J = 8.2, 1.6 Hz, 1H), 7.90–7.88 (m, 1H), 7.87–7.84 (m, 2H), 7.82 (dd, J = 8.6, 1.7 Hz, 1H), 7.76–7.69 (m, 2H), 7.56–7.50 (m, 2H), 7.04 (d, J = 8.1 Hz, 1H), 3.49 (d, J = 13.9 Hz, 6H).

[0125] 13 C NMR (150 MHz, CDCl 3 ):δ 188.89, 154.85, 144.34, 134.32, 134.02, 133.38, 132.51, 132.07, 130.47, 130.43, 128.67, 128.59, 127.77, 127.31, 126.75, 123.69, 123.37, 121.88, 107.47, 106.49, 27.40, 27.37.

[0126] HRMS-EI (m / z): C 22 H 18 N 2 O 2 of [M + H] + Calculated: 343.1441; Found: 343.1444.

[0127] (E)-5-(3-Cyclopropylacryloyl)-1,3-dimethyl-1,3-dihydro-2H-benzo[d]imidazol-2-one Preparation of (16)

[0128]

[0129] Dissolve the compound of formula (Ic) (0.1 mmol, 1.0 eq) and cyclopropanecarbaldehyde (0.1 mmol, 1.0 eq) in 1 mL of toluene, and reflux the reaction mixture at 120 °C overnight. Rotate evaporate to remove the solvent and concentrate the mixture, and separate the target product, namely compound 16, by column chromatography (dichloromethane∶methanol = 300∶1). The yield is 50.3%, and it is a pale yellow solid.

[0130] Use 1 H NMR and 13 C NMR to determine the structure of compound 16 and detect its molecular weight by HRMS-EI. The NMR spectra are as Figure 3 , and the data are as follows.

[0131] 1 H NMR (600 MHz, CDCl 3): δ 7.78 (d, J = 7.9 Hz, 1H), 7.65 (s, 1H), 7.10 (d, J = 15.0 Hz, 1H), 6.99 (d, J = 8.2 Hz, 1H), 6.58 (dd, J = 14.9, 10.4 Hz, 1H), 3.46 (d, J = 8.2 Hz, 6H), 1.68 (s, 1H), 1.03 (dd, J = 7.8, 2.5 Hz, 2H), 0.75 (dd, J = 4.6, 2.2 Hz, 2H).

[0132] 13 C NMR (150 MHz, CDCl 3 ): δ 188.39, 154.86, 154.48, 133.73, 131.89, 130.28, 123.13, 122.42, 107.41, 106.41, 27.35, 27.31, 15.32, 9.41, 9.11.

[0133] HRMS-EI (m / z): C 15 H 16 N 2 O 2 of [M + H] + Calculated: 257.1285; Found 257.1283.

[0134] (E)-5-(3-(Pyridin-2-yl)acryloyl)-1,3-dimethyl-1,3-dihydro-2H-benzo[d]imidazol-2-one Preparation of (22)

[0135]

[0136] The compound of formula (Ic) (0.1 mmol, 1.0 eq), pyridine-2-carboxaldehyde (0.1 mmol, 1.0 eq) were dissolved in 1 mL of toluene and refluxed at 120 °C overnight. The solvent was removed by rotary evaporation and the mixture was concentrated. The target product, i.e., compound 22, was separated by column chromatography (dichloromethane∶methanol = 200∶1) with a yield of 76%, as a yellow solid.

[0137] Using 1 H NMR and 13 C NMR to determine the structure of compound 22 and its molecular weight was detected by HRMS-EI. The NMR spectra are as Figure 4 , and the data are as follows.

[0138] 1 H NMR (600 MHz, CDCl 3):δ 8.71 (d, J = 4.7 Hz, 1H), 8.21 (d, J = 15.1 Hz, 1H), 7.99 (dd, J = 8.2, 1.6 Hz, 1H), 7.82 (d, J = 15.1 Hz, 1H), 7.79–7.75 (m, 2H), 7.50 (d, J = 7.7 Hz, 1H), 7.32 (dd, J = 7.6, 4.7 Hz, 1H), 7.04 (d, J = 8.2 Hz, 1H), 3.49 (d, J = 13.5 Hz, 6H).

[0139] 13 C NMR (150 MHz, CDCl 3 ):δ 188.91, 154.86, 153.27, 150.11, 142.19, 136.97, 134.26, 131.69, 130.39, 125.61, 125.28, 124.38, 123.89, 107.57, 106.59, 27.43.

[0140] HRMS-EI (m / z): C 17 H 15 N 3 O 2 of [M + H] + Calculated: 294.1237; Found: 294.1242.

[0141] (E)-5-(3-(Pyrimidin-2-yl)acryloyl)-1,3-dimethyl-1,3-dihydro-2H-benzo[d]imidazol-2-one Preparation of (28)

[0142]

[0143] The compound of formula (Ic) (0.1 mmol, 1.0 eq), pyrimidine-2-carbaldehyde (0.1 mmol, 1.0 eq) were dissolved in 1 mL of toluene and refluxed at 120 °C overnight. The solvent was removed by rotary evaporation and the mixture was concentrated. The target product, i.e., compound 28, was separated by column chromatography (dichloromethane∶methanol = 200∶1) with a yield of 91.1%, as a yellow solid.

[0144] Using 1 H NMR and 13 C NMR to determine the structure of compound 28 and the molecular weight was detected by HRMS-EI. The NMR spectra are as Figure 5 , and the data are as follows.

[0145] 1 H NMR (600 MHz, CDCl 3): δ 8.83 (d, J = 4.8 Hz, 2H), 8.36 (d, J = 15.4 Hz, 1H), 7.97 (dd, J = 8.2, 1.6 Hz, 1H), 7.84–7.76 (m, 2H), 7.26 (d, J = 1.7 Hz, 1H), 7.05 (d, J = 8.2 Hz, 1H), 3.49 (d, J = 10.5 Hz, 6H).

[0146] 13 C NMR (150 MHz, CDCl 3 ): δ 188.80, 163.43, 157.29, 154.85, 141.69, 134.44, 131.36, 130.37, 124.05, 120.19, 107.57, 106.61, 27.45, 27.42.

[0147] HRMS-EI (m / z): C 16 H 15 N 4 O 2 of [M + H] + Calculated: 295.1190; Found: 295.1190.

[0148] (E)-5-(3-(1-Methyl-1H-indol-3-yl)acryloyl)-1,3-dimethyl-1,3-dihydro-2H-benzo [d]imidazol-2-one (31) Preparation

[0149]

[0150] The compound of formula (Ic) (0.1 mmol, 1.0 eq), 1-methyl-1H-indole-3-carbaldehyde (0.1 mmol, 1.0 eq) were dissolved in 1 mL of toluene and refluxed at 120 °C overnight. The solvent was removed by rotary evaporation and the mixture was concentrated. The target product, namely compound 31, was separated by column chromatography (dichloromethane∶methanol = 100∶1). The yield was 89.5%, an orange-yellow solid.

[0151] Using 1 H NMR and 13 C NMR to determine the structure of compound 31 and the molecular weight was detected by HRMS-EI. The NMR spectra are as Figure 6 , and the data are as follows.

[0152] 1 H NMR (600 MHz, CDCl 3):δ 8.12 (d, J = 15.4 Hz, 1H), 8.06–8.02 (m, 1H), 7.92 (dd, J = 8.2, 1.6 Hz, 1H), 7.76 (d, J = 1.6 Hz, 1H), 7.62 (d, J = 15.4 Hz, 1H), 7.50 (s, 1H), 7.42–7.39 (m, 1H), 7.38–7.31 (m, 2H), 7.05 (d, J = 8.2 Hz, 1H), 3.86 (s, 3H), 3.50 (d, J = 13.8 Hz, 6H).

[0153] 13 C NMR (150 MHz, CDCl 3 ):δ 189.32, 154.92, 138.27, 138.15, 134.43, 133.49, 132.93, 130.31, 126.15, 123.15, 122.92, 121.50, 120.77, 116.71, 113.08, 110.14, 107.43, 106.44, 33.30, 27.40, 27.39.

[0154] HRMS-EI (m / z): C 21 H 20 N 3 O 2 of [M + H] + Calculated: 346.1550; Found: 346.1551.

[0155] ( E )-5-(3-(Thiophen-2-yl)acryloyl)-1,3-dimethyl-1,3-dihydro-2 H -benzo d imidazol-2-one Preparation of (34)

[0156]

[0157] The compound of formula (Ic) (0.1 mmol, 1.0 eq), thiophene-2-carbaldehyde (0.1 mmol, 1.0 eq) were dissolved in 1 mL of toluene and refluxed at 120 °C overnight. The solvent was removed by rotary evaporation and the mixture was concentrated. The target product, namely compound 34, was separated by column chromatography (dichloromethane∶methanol = 100∶1), with a yield of 33.6%, as an orange solid.

[0158] Using 1 H NMR and 13 C NMR to determine the structure of compound 34 and its molecular weight was detected by HRMS-EI. The NMR spectra are as Figure 7 , and the data are as follows.

[0159] 11H NMR (600 MHz, CDCl 3 ): δ 7.98 (d, J = 15.2 Hz, 1H), 7.85 (dd, J = 8.2, 1.6 Hz, 1H), 7.71 (d, J = 1.6 Hz, 1H), 7.44–7.36 (m, 3H), 7.10 (dd, J = 5.1, 3.6 Hz, 1H), 7.03 (d, J = 8.1 Hz, 1H), 3.49 (s, 3H), 3.47 (s, 3H).

[0160] 13 13C NMR (150 MHz, CDCl 3 ): δ 188.37, 154.88, 140.52, 136.72, 134.02, 131.94, 131.88, 130.43, 128.56, 128.34, 123.26, 120.53, 107.40, 106.51, 27.44, 27.40.

[0161] HRMS-EI (m / z): C 16 H 14 N 2 O 2 S of [M + H] + Calculated: 299.0849; Found: 299.0851.

[0162] 5-((2E,4E)-5-Phenylpenta-2,4-dienoyl)-1,3-dimethyl-1,3-dihydro-2H-benzo[d]imidazol- 2-one (50) Preparation

[0163]

[0164] The compound of formula (Ic) (0.1 mmol, 1.0 eq) and cinnamaldehyde (0.1 mmol, 1.0 eq) were dissolved in 1 mL of toluene and refluxed at 120 °C overnight. The solvent was removed by rotary evaporation and the mixture was concentrated. The target product, compound 50, was separated by column chromatography (dichloromethane). The yield was 24.8%, an orange-yellow solid.

[0165] Using 1 1H NMR and 13 13C NMR to determine the structure of compound 50 and its molecular weight was detected by HRMS-EI. The NMR spectra are as Figure 8 , and the data are as follows.

[0166] 1 1H NMR (600 MHz, CDCl 3):δ 7.83 (d, J = 8.1 Hz, 1H), 7.72–7.61 (m, 2H), 7.51 (d, J = 7.5 Hz, 2H), 7.38 (t, J = 7.5 Hz, 2H), 7.33 (d, J = 7.1 Hz, 1H), 7.18 (d, J = 14.8 Hz, 1H), 7.03 (d, J = 11.1 Hz, 3H), 3.48 (d, J = 10.1 Hz, 6H).

[0167] 13 C NMR (150 MHz, CDCl 3 ):δ 188.91, 154.86, 144.27, 141.63, 136.17, 133.92, 132.07, 130.39, 129.14, 128.83, 128.61, 127.22, 126.99, 126.21, 125.14, 123.19, 107.38, 106.48, 27.38, 27.33.

[0168] HRMS-EI (m / z): C 20 H 18 N 2 O 2 of [M + H] + Calculated: 319.1441; Found: 319.1436.

[0169] Preparation of the Compound of Formula (II) in Production Example 2

[0170] In some embodiments, the method for preparing the compound of formula (II) is as shown in formula (ii).

[0171]

[0172] The method for preparing the compound of formula (II) may include:

[0173] - Preparing the compound of formula (II) from the compound of formula (IIa) (acetophenone).

[0174]

[0175] 2.1 Preparing the compound of formula (II) from the compound of formula (IIa)

[0176] Dissolve the compound of formula (IIa) (0.3 mmol, 1.0 eq) and sodium hydroxide (1.2 mmol, 4.0 eq) in 3 mL of anhydrous methanol, and react at room temperature for 15 minutes. Add R 1 –CHO (0.3 mmol, 1.0 eq), and react overnight at room temperature. After the reaction is complete, extract with ethyl acetate, remove water, evaporate to dryness, and separate by column chromatography to obtain the compound of formula (II).

[0177] The various compounds of formula (II) prepared in this production example are shown as compound 54 and compound 55 below.

[0178]

[0179] Preparation of the Compounds of Formula (III) and Formula (III′) in Production Example 3

[0180] In some embodiments, the preparation methods of the compound of formula (III) and the compound of formula (III') are as shown in formula (iii).

[0181]

[0182] The preparation methods of the compound of formula (III) and the compound of formula (III') may include:

[0183] - preparing the compound of formula (IIIc) (N-(4-(2-(triphenyl-λ 5 - phosphoniumyl)acetyl)phenyl)acetamide) from the compound of formula (IIIb) (N-(4-(2-chloroacetyl)phenyl)acetamide);

[0184] - preparing the compound of formula (III) from the compound of formula (IIIc); and

[0185] - optionally, preparing the compound of formula (III') from the compound of formula (III).

[0186]

[0187] 3.1 Preparing the compound of formula (IIIc) from the compound of formula (IIIb)

[0188] Weigh the compound of formula (IIIb) and triphenylphosphine and place them in a three-necked flask. Under argon protection, add dry acetonitrile solvent to it with a syringe, and place the reaction flask in an oil bath at 83 °C for overnight reaction, where the equivalent ratio of the compound of formula (IIIb) to triphenylphosphine is 1:1.1.

[0189] Monitor the reaction progress by TLC. After the reaction is complete, cool it to room temperature, add saturated sodium carbonate solution to quench the reaction, extract with ethyl acetate, remove water, spin dry, and separate by column chromatography to obtain the compound of formula (IIIc).

[0190] 3.2 Preparing the compound of formula (III) from the compound of formula (IIIc)

[0191] Dissolve the compound of formula (IIIc) (0.1 mmol, 1.0 eq) and R1–CHO (0.1 mmol, 1.0 eq) in 3 mL of toluene, and reflux at 120 °C for overnight reaction. Rotate and evaporate to remove the solvent, and purify by column chromatography to obtain the compound of formula (III).

[0192] The various compounds of formula (III) prepared in this Preparation Example are shown as Compound 56 below, but are not limited thereto.

[0193]

[0194] 3.3 Preparation of the compound of formula (III′) from the compound of formula (III)

[0195] Dissolve the compound of formula (III) (0.1 mmol, 1.0 eq) in 1 mL of N,N-dimethylformamide (DMF), add sodium hydride (0.4 mmol, 4.0 eq) under an ice bath, stir for 15 minutes, then slowly add methyl iodide (0.4 mmol, 4.0 eq) dropwise, and react at room temperature for 12 hours. Monitor the reaction progress by TLC. After the reaction is complete, extract with ethyl acetate, remove water, evaporate to dryness, and separate by column chromatography to obtain the compound of formula (III′).

[0196] The various compounds of formula (III′) prepared in this Preparation Example are shown as Compound 57 below.

[0197]

[0198] Preparation of (E)-N-(4-(3-(4-Ethynylphenyl)acryloyl)phenyl)-N-methylacetamide (57)

[0199] Dissolve the compound of formula (III) (where R 1 is 4-ethynylphenyl; i.e., (E)-N-(4-(3-(4-ethynylphenyl)acryloyl)phenyl)acetamide) (0.1 mmol, 1.0 eq) in 1 mL of DMF, add sodium hydride (0.4 mmol, 4.0 eq) under an ice bath, stir for 15 minutes, then slowly add methyl iodide (0.4 mmol, 4.0 eq) dropwise, and react at room temperature for 12 hours. Monitor the reaction progress by TLC. After the reaction is complete, extract with ethyl acetate, remove water, evaporate to dryness, and perform column chromatography (dichloromethane) separation to obtain the target product, i.e., Compound 57. The yield is 62.6%, and it is a yellow solid.

[0200] Using 1 1H NMR and 13 13C NMR to determine the structure of Compound 57 and detect the molecular weight by HRMS-EI. The NMR spectra are as Figure 9 follows, and the data are as follows.

[0201] 1 1H NMR (600 MHz, CDCl 3):δ 8.08 (d, J = 8.1 Hz, 2H), 7.81 (d, J = 15.6 Hz, 1H), 7.61 (d, J = 8.0 Hz, 2H), 7.56–7.53 (m, 2H), 7.51 (s, 1H), 7.34 (d, J = 8.0 Hz, 2H), 3.32 (s, 3H), 3.22 (s, 1H), 1.96 (s, 3H).

[0202] 13 C NMR (150 MHz, CDCl 3 ):δ 188.92, 148.43, 144.56, 144.13, 134.99, 132.78, 132.66, 132.03, 129.94, 128.28, 127.43, 127.02, 124.42, 122.39, 121.69, 83.08, 42.62, 22.52.

[0203] HRMS-EI (m / z): C 20 H 17 NO 2 of [M + H] + Calculated: 304.1332; Found: 304.1331.

[0204] Preparation of the Compound of Formula (IV) in Production Example 4

[0205] In some embodiments, the method for preparing the compound of formula (IV) is as shown in formula (iv).

[0206]

[0207] The method for preparing the compound of formula (IV) may include:

[0208] - Preparing the compound of formula (IV) (N-benzyl-1,3-dimethyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazole-5-sulfonamide) from the compound of formula (IVa) (1,3-dimethyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazole-5-sulfonyl chloride).

[0209]

[0210] 4.1 Preparation of the compound of formula (IV) from the compound of formula (IVa)

[0211] Under argon protection, dissolve the compound of formula (IVa) (4.1 mmol, 1.1 eq) in 30 mL of dry DCM solution, and then successively add triethylamine (5.6 mmol, 1.5 eq) and benzylamine (3.73 mmol, 1.0 eq), and react at room temperature for 12 hours. Monitor the reaction progress by TLC, perform conventional water removal operations after extraction with dichloromethane, spin-dry and then perform column chromatography separation to obtain the compound of formula (IV).

[0212] The compound of formula (IV) prepared in this application is shown as Compound 58 below.

[0213]

[0214] Preparation of N-Benzyl-1,3-dimethyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazole-5-sulfonamide (58)

[0215] Under argon protection, dissolve the compound of formula (IVa) (4.1 mmol, 1.1 eq) in 30 mL of dry DCM solution, and then successively add triethylamine (5.6 mmol, 1.5 eq) and benzylamine (3.73 mmol, 1.0 eq), and react at room temperature for 12 hours. Monitor the reaction progress by TLC. After the reaction is complete, extract with dichloromethane, remove water, spin-dry, and perform column chromatography (dichloromethane∶methanol = 100∶1) separation to obtain the target product, that is, Compound 58. The yield is 14.5%, a white solid.

[0216] Use 1 1H NMR and 13 13C NMR to determine the structure of Compound 58 and detect the molecular weight by HRMS-EI. The NMR spectra are as Figure 10 , and the data are as follows.

[0217] 1 1H NMR (600 MHz, CDCl 3 3): δ 7.68 (dd, J = 8.2, 1.7 Hz, 1H), 7.44 (d, J = 1.7 Hz, 1H), 7.28–7.26 (m, 1H), 7.26–7.24 (m, 2H), 7.21–7.18 (m, 2H), 7.05 (d, J = 8.2 Hz, 1H), 4.68 (t, J = 6.1 Hz, 1H), 4.15 (d, J = 6.2 Hz, 2H), 3.47 (s, 3H), 3.44 (s, 3H).

[0218] 13 13C NMR (150 MHz, CDCl 3):δ 154.69, 136.16, 133.36, 132.57, 130.08, 128.67, 127.95, 127.87, 121.26, 107.02, 106.38, 47.33, 27.48, 27.45。

[0219] HRMS - EI (m / z): C 16 H 17 N 3 O 3 Calculated value of [M + H]+ for S: 332.1063; Measured value: 332.1061.

[0220] Evaluation of Biological Activity (Inhibitory Efficiency of CYP1A1 Activity) in Experimental Example 1

[0221] Cell Culture

[0222] Human Caco - 2 cells and mouse RAW264.7 cells overexpressing CYP1A1 (CYP1A1hiRAW264.7 cells, hereinafter referred to as CY h RAW cells) were used for cell culture and the following experiments. According to the method disclosed in CN117925529A, RAW264.7 cells were transfected with CYP1A1 - overexpressing virus to obtain CYP1A1 - overexpressing RAW264.7 cells.

[0223] Cell passage

[0224] Passage was carried out when the cell density reached 80%. Discard the cell culture supernatant, wash twice with sterile PBS for 2 minutes each time. Add 2 mL of trypsin to the cell culture flask and digest at 37°C for 2 minutes. Add twice the volume of complete medium to terminate digestion. Centrifuge at 1200 rpm for 3 minutes, discard the supernatant, and resuspend the cells with 2 mL of culture medium. Add the cell suspension to a new cell culture flask at a ratio of 1:3, add 4 mL of complete medium, and culture under the conditions of 37°C and 5% CO 2 conditions.

[0225] Cell cryopreservation

[0226] Discard the cell culture supernatant, wash twice with sterile PBS for 2 minutes each time. After trypsin digestion and centrifugation, resuspend the cells with serum - free cell cryopreservation solution. Cryopreserve at a concentration of 1×10 6 –1×10 7 viable cells per 1 mL of serum - free cell cryopreservation solution, place in an - 80°C refrigerator for 24 hours and then transfer to liquid nitrogen for storage.

[0227] Cell recovery

[0228] Remove the cryotube from liquid nitrogen, thaw it in a 37°C water bath, add it to a centrifuge tube containing 2 mL of complete medium, mix well, centrifuge at 1200 rpm for 3 minutes, discard the supernatant, resuspend the cells in 5 mL of complete medium, add them to a cell culture flask, and culture at 37°C and 5% CO 2 conditions.

[0229] 1.1 EROD assay

[0230] 7-Ethoxy-3H-phenoxazin-3-one O-deethylase (EROD) is the expression product of CYP1A1. CYP1A1 inhibitors inhibit the activity of CYP1A1, thereby inhibiting the activity of EROD; therefore, detecting the degree of inhibition of EROD activity can detect the degree of inhibition of CYP1A1 activity by CYP1A1 inhibitors.

[0231] Inoculate 500 μL of Caco-2 cell suspension or CY h RAW cell suspension into a 48-well plate, inoculate 5×10 4 cells per well, and allow them to adhere for 4 hours.

[0232] Prepare DMSO solutions of each drug at each concentration (the compounds prepared in each production example, or bergapten (bergapten) as a positive control, or no compound added as a negative control), and dilute them 10-fold with DMEM medium to obtain the drug solutions of each concentration.

[0233] Add 5 μL of the drug solutions of each concentration to the 48-well plate inoculated with cells, so that the final concentration of each drug is 5 μM, 500 nM, or 50 nM, to pretreat the cells for 12 hours; set 3 replicates for each drug concentration.

[0234] After pretreatment, discard the supernatant, wash the cells twice with warm PBS, and add 100 μL of sterile PBS containing 5 μM of 7-ethoxy-3H-phenoxazin-3-one and 10 μM of dicoumarol to each well. Culture at 37°C and 5% CO 2 conditions for 30 minutes. Aspirate 75 μL of the culture supernatant into a black-bottom 96-well plate, and add 125 μL of anhydrous methanol to each well to terminate the reaction.

[0235] Detect with a fluorescence microplate reader (excitation light 530 nm, emission light 590 nm).

[0236] Test Results

[0237] Perform the EROD assay with Compounds 1 to 58 of the present application, and the results are shown in Table 1.

[0238] Table 1

[0239]

[0240]

[0241]

[0242] 1.2 Structure-Activity Relationship Study

[0243] Figure 11 The components of the compound of formula (I) are shown, and the compound of formula (I) can be regarded as including part A (R 1 part), part B (acrylylidene part) and part C (1,3-dimethyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl part). Based on the experimental results of the above Experimental Example 1.1, the effects of the structural selection of part A, part B, and part C on the inhibitory activity of the compound of formula (I) against CYP1A1 will be discussed below.

[0244] Structure-Activity Relationship Study of Part C

[0245] To understand the effect of part C on the inhibitory activity of the compound against CYP1A1, the compound of formula (I) was compared with the compounds of formula (II), formula (III), and formula (III′) which differ from it in part C (i.e., the same R 1 ) for the inhibitory activity against CYP1A1.

[0246] Comparing Compound 1 (the compound of formula (I), R 1 is 4-fluorophenyl) and Compound 50 (the compound of formula (II), R 1 is 4-fluorophenyl), it can be seen that the inhibitory activity of Compound 1 against CYP1A1 is significantly greater than that of Compound 50 against CYP1A1 (i.e., more effectively inhibits CYP1A1).

[0247] Comparing Compound 20 (the compound of formula (I), R 1 is 4-ethynylphenyl) and Compound 54 (the compound of formula (III), R 1 is 4-ethynylphenyl), Compound 55 (the compound of formula (III′), R 1 is 4-ethynylphenyl), and Compound 56 (the compound of formula (II), R 1 is 4-ethynylphenyl), it can be seen that the inhibitory activity of Compound 20 against CYP1A1 is significantly greater than that of Compound 54, Compound 55, and Compound 56 against CYP1A1, especially the inhibitory activity against CYP1A1 in Caco-2 cells.

[0248] This indicates that part C is very important for the inhibitory activity against CYP1A1.

[0249] Structure-Activity Relationship Study of Part B

[0250] To understand the effect of part B on the inhibitory activity of the compound against CYP1A1, the inhibitory activities of the compound of formula (I) and the compound of formula (IV) which differs from it only in part B against CYP1A1 were compared.

[0251] Comparing compound 17 (the compound of formula (I), R 1 being phenyl) and compound 58 (the compound of formula (IV) having the corresponding phenyl moiety), it can be seen that the inhibitory activity of compound 17 against CYP1A1 is significantly greater than that of compound 58 against CYP1A1.

[0252] This indicates that part B is also important for the inhibitory activity against CYP1A1.

[0253] Structure-Activity Relationship Study of Part A

[0254] To understand the effect of part A on the inhibitory activity of the compound against CYP1A1, the inhibitory activities of the compounds of formula (I) with different R 1 against CYP1A1 were compared.

[0255] Based on R 1 being phenyl, halogen groups were introduced

[0256] At different positions on the phenyl group of compound 17 (R 1 being phenyl), fluorine was introduced respectively to obtain compounds 46 (R 1 being 2-fluorophenyl), compound 47 (R 1 being 3-fluorophenyl), and compound 1 (R 1 being 4-fluorophenyl). It can be seen that whether fluorine is introduced at the ortho, meta, or para position, the inhibitory activity against CYP1A1 is very significantly enhanced. Among them, the enhancement of the inhibitory activity against CYP1A1 in Caco-2 cells by introducing fluorine at the ortho and meta positions (compound 46, compound 47) is more significant, while the enhancement of the inhibitory activity against CYP1A1 in RAW cells by introducing fluorine at the para position (compound 1) is more significant. 1 ... h ...

[0257] Based on R 1 being monofluorophenyl, more fluorine atoms were further introduced, such as compound 4 (R 1 being 3,5-difluorophenyl), compound 5 (R 1 being 2,4-difluorophenyl), compound 48 (R 1is 2,4,5-trifluorophenyl), Compound 49 (R 1 is 2,3,4,5,6-pentafluorophenyl), compared with R 1 being compounds with a monofluorophenyl group (Compound 46, Compound 47, Compound 1), there is no further improvement in the inhibitory activity against CYP1A1. This indicates that the inhibitory activity of the compounds of formula (I) against CYP1A1 does not increase with the increase in the number of fluorine atoms in R 1 in it.

[0258] On the basis of R 1 being a monofluorophenyl group, introducing other halogens, such as Compound 3 (R 1 is 2-chloro-4-fluorophenyl), there is also no further improvement in the inhibitory activity against CYP1A1. However, on the basis of R 1 being a monofluorophenyl group, introducing a boronic acid group, such as Compound 2 (R 1 is 4-borono-2-fluorophenyl), has a good effect on the inhibitory activity of CYP1A1 in Caco-2. Changing to introduce a group with a relatively large electronegativity such as trifluoromethoxy, such as Compound 6 (R 1 is 3-(trifluoromethoxy)phenyl), it is found that its inhibitory activity against CYP1A1 has a certain improvement.

[0259] On the basis of R 1 being a phenyl group, introducing an electron-withdrawing group and an electron-donating group

[0260] On the phenyl group of Compound 17 (R 1 is a phenyl group), introducing an electron-withdrawing group or an electron-donating group, to obtain Compound 7 (R 1 is 4-methylthiophenyl), Compound 8 (R 1 is 4-butoxyphenyl), Compound 9 (R 1 is 4-tert-butylphenyl), Compound 10 (R 1 is 4-(tert-butyldimethylsilyloxy)phenyl), Compound 11 (R 1 is 4-(dimethylamino)phenyl), Compound 12 (R 1 is 4-phenylphenyl), Compound 15 (R 1 is 2-(diphenylphosphino)phenyl), Compound 18 (R 1 is 4-nitrophenyl), Compound 19 (R 1 is 4-cyanophenyl), Compound 43 (R 1 is 2-ethynylphenyl), Compound 44 (R 1 is 3-ethynylphenyl), Compound 20 (R 1 is 4-ethynylphenyl).

[0261] It can be seen that compounds 10 and 11 exhibit good inhibitory activity against CYP1A1 in Caco-2 cells, while compounds 12 and 15 have good inhibitory activity against CYP1A1 at a higher concentration (5 μM).

[0262] Compounds 43, 44, and 20 with alkynyl substituents exhibit particularly excellent inhibitory activity against CYP1A1 in various cell lines. Among them, the inhibitory activity of compounds 43 and 44 against CYP1A1 is significantly higher than that of the positive control (bergapten). The inhibitory activity of compound 43 against CYP1A1 is the best among them.

[0263] R 1 is other aryl or cycloalkyl

[0264] Select other aryl or cycloalkyl as R 1 , to obtain compound 13 (R 1 is naphthyl), compound 14 (R 1 is anthryl), compound 16 (R 1 is cyclopropyl).

[0265] It can be seen that compound 16 has good inhibitory activity against CYP1A1 at a higher concentration (5 μM).

[0266] R 1 is a heterocyclic nitrogen

[0267] Select pyridyl as R 1 , to obtain compound 22 (R 1 is pyridin-2-yl), compound 23 (R 1 is pyridin-3-yl), compound 24 (R 1 is pyridin-4-yl). It can be seen that the inhibitory activity of compound 22 against CYP1A1 is better than that of compound 23 and compound 24. On the basis of compound 23 (R 1 is pyridin-3-yl), further introduce substituents to obtain compound 25 (R 1 is 6-methylpyridin-3-yl), compound 26 (R 1 is 2-bromopyridin-3-yl), and the inhibitory activity against CYP1A1 is significantly enhanced, especially for compound 25.

[0268] Select pyrrolyl as R 1 and introduce substituents to obtain compound 21 (R 1 is 1-(tert-butoxycarbonyl)-1H-pyrrol-2-yl) and compound 27 (R 1 is 1-methyl-1H-pyrrol-2-yl). Compound 21 exhibits good inhibitory activity against CY hThe inhibitory activity of CYP1A1 in RAW cells, while compound 27 showed good inhibitory activity of CYP1A1 in Caco-2 cells.

[0269] The quinolinyl group was selected as R 1 , to obtain compound 29 (R 1 is quinolin-2-yl), compound 45 (R 1 is quinolin-3-yl). Both compound 29 and compound 45 showed good inhibitory activity against CYP1A1.

[0270] The indolyl group was selected as R 1 , to obtain compound 30 (R 1 is 1-acetyl-1H-indol-3-yl), compound 31 (R 1 is 1-methyl-1H-indol-3-yl), compound 32 (R 1 is 1-benzyl-1H-indol-3-yl), compound 45 (R 1 is 1-benzyl-5-methoxy-1H-indol-3-yl). Compound 30 and compound 31 showed good inhibitory activity against CYP1A1, and compound 32 showed good inhibitory activity of CYP1A1 in Caco-2 cells.

[0271] R 1 is an oxacycle or a thiacycle

[0272] The furyl group was selected as R 1 , to obtain compound 37 (R 1 is furan-2-yl), compound 36 (R 1 is furan-3-yl); the benzofuryl group was selected as R 1 , to obtain compound 39 (R 1 is benzo[b]furan-2-yl), compound 38 (R 1 is benzo[b]furan-3-yl). Compound 36 showed good inhibitory activity of CYP1A1 in Caco-2 cells, while compound 37 showed good inhibitory activity of CYP1A1 in CY h RAW cells. This result was also correspondingly reflected in compound 38 and compound 39.

[0273] The thienyl group was selected as R 1 , to obtain compound 34 (R 1 is thien-2-yl); the benzothienyl group was selected as R 1 , to obtain compound 35 (R 1 is benzo[b]thiophen-2-yl). Compound 34 showed obvious inhibitory activity against CYP1A1.

[0274] Insert vinylidene

[0275] Insert vinylidene into Compound 17 (R 1 is phenyl) and Compound 1 (R 1 is 4-fluorophenyl) to obtain Compound 50 (R 1 is (E)-styryl) and Compound 51 (R 1 is (E)-4-fluorostyryl).

[0276] Compared with Compound 17 and Compound 1 respectively, Compound 50 and Compound 51 did not show better inhibitory activity against CYP1A1.

[0277] 1.3 IC 50 value determination

[0278] According to the above EROD test, but the final concentration of each drug was changed to 10 pM, 100 pM, 1 nM, 10 nM, 100 nM, 1 μM, 10 μM, 20 μM to determine the IC 50 value.

[0279] Please refer to Figure 12 and Figure 13 which show the EROD enzyme activity intensity obtained by performing the EROD test using Caco-2 and CY 1 RAW cells respectively with Compound 1 (R 1 is 4-fluorophenyl), Compound 6 (R 1 is 3-(trifluoromethoxy)phenyl), Compound 20 (R 1 is 4-ethynylphenyl), Compound 29 (R 1 is quinolin-2-yl), Compound 34 (R 1 is thiophen-2-yl), Compound 25 (R 1 is 6-methylpyridin-3-yl), Compound 26 (R 1 is 2-bromopyridin-3-yl), Compound 31 (R 1 is 1-methyl-1H-indol-3-yl), Compound 43 (R 1 is trimethylsilylethynyl), and Compound 53 (R h is trimethylsilylethynyl). Table 2 shows the calculated IC h values of each compound in Caco-2 and CY 50 RAW cells.

[0280] Table 2

[0281]

[0282] 1.4 Re-verification of the inhibitory effect of CYP1A1 activity of some preferred compounds

[0283] Evaluation of CYP1A1 Activity Inhibitory Effect

[0284] Using human Caco-2 cells and murine CY h RAW cells, compound 43 (R 1 is 2-ethynylphenyl) and compound 31 (R 1 is 1-methyl-1H-indol-3-yl) were selected, and bergapten was used as a positive control, and this verification was carried out using the aforementioned EROD assay at the cellular level.

[0285] The results are as Figure 14 . Figure 14 The results showed that compound 43 (R 1 is 2-ethynylphenyl) and compound 31 (R 1 is 1-methyl-1H-indol-3-yl) effectively inhibited CYP1A1 activity.

[0286] Pharmacodynamic Evaluation of the Compound's Ability to Enhance the Body's Anti-Bacterial Ability in Experimental Example 2

[0287] Grouping of Experimental Animals

[0288] Forty 7-week-old male C57 / 6J mice, each weighing approximately 20 g, were selected. They were randomly divided into four groups (vehicle control group, compound 43 group, compound 31 group, bergapten group), with 10 mice in each group; bergapten was used as the positive control drug.

[0289] - Vehicle control group: A mixture of 10% (v / v) DMSO and 90% (v / v) corn oil (vehicle) was administered during dosing.

[0290] - Compound 43 group: The vehicle solution of compound 43 was administered during dosing at a dose of 25 mg of compound 43 per kg of mouse body weight.

[0291] - Compound 31 group: The vehicle solution of compound 31 was administered during dosing at a dose of 25 mg of compound 31 per kg of mouse body weight.

[0292] - Bergapten group: The vehicle solution of bergapten was administered during dosing at a dose of 25 mg of bergapten per kg of mouse body weight.

[0293] Drug Preparation

[0294] - Vehicle control group: 200 μl of DMSO was added to 1.8 mL of corn oil and vortexed to obtain the vehicle.

[0295] - Compound 43 group: 5 mg of compound 43 was dissolved in 200 μl of DMSO, and corn oil was added to 2 mL and vortexed to obtain the compound 43 solution.

[0296] - Compound 31 group: 5 mg of compound 31 was dissolved in 200 μl of DMSO, and corn oil was added to make 2 mL. The mixture was vortexed to obtain the compound 31 solution.

[0297] - Bergapten group: 5 mg of bergapten was dissolved in 200 μl of DMSO, and corn oil was added to make 2 mL. The mixture was vortexed to obtain the bergapten solution.

[0298] Drug Administration

[0299] - Solvent control group: 200 μl of solvent was intraperitoneally injected.

[0300] - Compound 43 group: 200 μl of the prepared compound 43 solution was intraperitoneally injected.

[0301] - Compound 31 group: 200 μl of the prepared compound 31 solution was intraperitoneally injected.

[0302] - Bergapten group: 200 μl of the prepared bergapten solution was intraperitoneally injected.

[0303] The mice after the above administration were used in the following Experimental Examples 2.1–2.5.

[0304] 2.1 Pharmacodynamic evaluation of the compound against methicillin-resistant Staphylococcus aureus (MRSA) infection

[0305] Animal Model Establishment

[0306] 12 hours after the mice were administered, 1.5×10 8 CFU of MRSA was intraperitoneally injected into the mice.

[0307] Results

[0308] The survival of the mice was observed, and the results are shown in Figure 15 (a). It can be seen that the compound of the present application can enhance the ability of the body to resist MRSA infection.

[0309] 2.2 Pharmacodynamic evaluation of the compound against Acinetobacter baumannii (AB) infection

[0310] Animal Model Establishment

[0311] 12 hours after the mice were administered, 1×10 8 CFU of AB was intraperitoneally injected.

[0312] Results

[0313] The survival of the mice was observed, and the results are shown in Figure 15In (b). It can be seen that the compounds of the present application can enhance the body's ability to resist AB infection.

[0314] 2.3 Pharmacodynamic evaluation of the compound in the treatment of cecal ligation and puncture (CLP) infection

[0315] Cecal ligation and puncture (CLP) is one of the best animal models for replicating clinical sepsis models. It can well reflect the two-way changes in immunology and hemodynamics during the development of sepsis. Ligation of the distal cecum causes tissue necrosis. After perforation, the intestinal contents mixed with bacteria continuously leak into the abdominal cavity, causing infection, thereby inducing peritonitis, leading to intestinal bacterial translocation, activating the immune system, and ultimately resulting in respiratory and circulatory system failure, as well as septic shock and multiple organ dysfunction syndrome, until death.

[0316] Animal Model Establishment

[0317] Twelve hours after the mice were administered the drug, a cecal ligation and puncture (CLP) model was established. The mice were anesthetized by intraperitoneal injection of 10% chloral hydrate (20 mg / kg). After disinfecting the abdomen, a midline laparotomy was performed. The root of the cecum was ligated with silk thread, and the distal 1.5 cm to 1.8 cm of the cecum was ligated with sterile silk thread. The ligated distal cecum was punctured through the center with an 18-gauge sterile needle. After squeezing out a small amount of contents, the cecum was gently pushed back into the abdominal cavity and the incision was sutured. The vehicle control group only underwent laparotomy, abdominal closure, and resuscitation, without cecal ligation and puncture.

[0318] Results

[0319] The survival of the mice was observed, and the results are shown in Figure 15 In (c). It can be seen that the compounds of the present application have an obvious therapeutic effect on mice infected with CLP.

[0320] 2.4 The compound enhances the body's ability to clear MRSA

[0321] Animal Model Establishment

[0322] Twelve hours after the mice were administered the drug, 5×10 7 CFU of MRSA was injected intraperitoneally. Twelve hours later, the mice were anesthetized and sacrificed by cervical dislocation. The whole body was soaked in 75% alcohol for disinfection. The abdominal wall of the mice was cut open, 5 mL of pre-cooled normal saline and 1 mL of air were injected, and the abdomen was massaged to make the liquid flow fully in the abdominal cavity. The peritoneal lavage fluid of the mice was aspirated into a sterile 15 mL centrifuge tube, diluted 10,000 times with sterile normal saline, and then 20 μL of the diluted peritoneal lavage fluid was added to the blood agar plate medium and cultured at 37 °C for 24 hours.

[0323] Results

[0324] Take pictures and perform colony counting. The results are shown in Figure 16 (a) and (b) of

[0325] 2.5 Compounds enhance the body's ability to clear AB

[0326] Animal Model Establishment

[0327] After 12 hours of drug administration to the mice, inject 2.5×10 7 CFU of AB intraperitoneally. After 12 hours, anesthetize the mice and decapitate them. Immerse the whole body of the mice in 75% alcohol for disinfection. Cut open the abdominal wall of the mice, inject 5 mL of pre-cooled physiological saline and 1 mL of air, and massage the abdomen to make the liquid flow fully in the abdominal cavity. Aspirate the peritoneal lavage fluid of the mice into a sterile 15 mL centrifuge tube, dilute it 10,000 times with sterile physiological saline, and then aspirate 20 μL of the diluted peritoneal lavage fluid and add it to the blood agar plate medium, and culture it at 37 °C for 24 hours.

[0328] Results

[0329] Take pictures and perform colony counting. The results are shown in Figure 16 (c) and (d) of

[0330] Pharmacodynamic Evaluation of the Compound's Protective Effect on Liver and Kidney Injury Caused by Bacterial Infection in Experimental Example 3

[0331] Grouping of Experimental Animals

[0332] Select 40 male C57 / 6J mice at 7 weeks of age, and the weight of each mouse is about 20 g. Randomly divide them into four groups (blank control group, vehicle control group, compound 43 group, compound 31 group), with 10 mice in each group.

[0333] - Blank control group: Administer physiological saline during drug administration.

[0334] - Vehicle control group: Administer a mixture of 10% (v / v) DMSO and 90% (v / v) corn oil (vehicle) during drug administration.

[0335] - Compound 43 group: Administer the vehicle solution of compound 43 during drug administration, and the dose is 25 mg of compound 43 per kg of mouse body weight.

[0336] - Compound 31 group: Administer the vehicle solution of compound 31 during drug administration, and the dose is 25 mg of compound 31 per kg of mouse body weight.

[0337] Drug Preparation

[0338] - Solvent control group: Take 200 μl of DMSO and add 1.8 mL of corn oil, then vortex to mix evenly to obtain the solvent.

[0339] - Compound 43 group: Dissolve 5 mg of compound 43 in 200 μl of DMSO, add corn oil to make up to 2 mL, and vortex to mix evenly to obtain the compound 43 solution.

[0340] - Compound 31 group: Dissolve 5 mg of compound 31 in 200 μl of DMSO, add corn oil to make up to 2 mL, and vortex to mix evenly to obtain the compound 31 solution.

[0341] Drug Administration

[0342] - Blank control group: Intraperitoneally inject 200 μl of normal saline.

[0343] - Solvent control group: Intraperitoneally inject 200 μl of the solvent.

[0344] - Compound 43 group: Intraperitoneally inject 200 μl of the prepared compound 43 solution.

[0345] - Compound 31 group: Intraperitoneally inject 200 μl of the prepared compound 31 solution.

[0346] Animal Model Establishment

[0347] 12 hours after drug administration to the mice, intraperitoneally inject 5×10 7 CFU of MRSA (the blank control group injects the blank solution). 24 hours later, intraperitoneally inject 10% chloral hydrate (20 mg / kg) to anesthetize the mice. Use an ophthalmic forceps to remove one eyeball of the mice, collect the blood into a coagulation-promoting tube, and let it stand at room temperature for 4 hours. Then centrifuge at 3000 rpm for 10 minutes, carefully aspirate the supernatant into a sterile 1.5 mL centrifuge tube, and store it in a -80°C refrigerator as the test sample for Experimental Examples 3.1 - 3.2.

[0348] 3.1 Pharmacodynamic evaluation of the protective effect of the compound on liver injury caused by bacterial infection

[0349] Use the determination of aspartate aminotransferase (AST; also known as GOT) and alanine aminotransferase (ALT; also known as GPT) to detect the liver function of the mice.

[0350] AST Determination

[0351] The AST assay kit was purchased from Nanjing Jiancheng Bioengineering Research Institute Co., Ltd., product number: C010 - 2 - 1.

[0352] Add 20 μL of pre-warmed AST substrate solution at 37°C to both the measurement group and the control group, and add 5 μL of the sample to be tested to the measurement group. React at 37°C for 30 minutes. Then add 20 μL of 2,4-dinitrophenylhydrazine solution to both the measurement group and the control group, and add 5 μL of the sample to be tested to the control group. React at 37°C for 20 minutes. Then add 200 μL of 0.4 M aqueous sodium hydroxide solution to both the measurement group and the control group, gently shake horizontally to mix evenly, and let stand at room temperature for 15 minutes. Measure the OD values of the measurement group and the control group at a wavelength of 510 nm using an enzyme-linked immunosorbent assay reader. Calculate the corresponding AST activity units (in Karman units (karU)) according to the standard curve based on the absolute OD value (the OD value of the measurement group minus the OD value of the control group).

[0353] ALT Determination

[0354] The ALT assay kit was purchased from Nanjing Jiancheng Bioengineering Research Institute Co., Ltd., product number: C009-2-1.

[0355] Add 20 μL of pre-warmed ALT substrate solution at 37°C to both the measurement group and the control group, and add 5 μL of the sample to be tested to the measurement group. React at 37°C for 30 minutes. Then add 20 μL of 2,4-dinitrophenylhydrazine solution to both the measurement group and the control group, and add 5 μL of the sample to be tested to the control group. React at 37°C for 20 minutes. Then add 200 μL of 0.4 M aqueous sodium hydroxide solution to both the measurement group and the control group, gently shake horizontally to mix evenly, and let stand at room temperature for 15 minutes. Measure the OD values of the measurement group and the control group at a wavelength of 510 nm using an enzyme-linked immunosorbent assay reader. Calculate the corresponding ALT activity units (in Karman units (karU)) according to the standard curve based on the absolute OD value (the OD value of the measurement group minus the OD value of the control group).

[0356] Results

[0357] The AST measurement results are as shown in Figure 17 of (a); the ALT measurement results are as shown in Figure 17 of (b). It can be seen that the compound of the present application can effectively reduce the AST level and ALT level of liver injury caused by bacterial infection.

[0358] According to the test results of AST measurement and ALT measurement, it can be known that the compound of the present application has a certain protective effect on liver injury caused by bacterial infection.

[0359] 3.2 Pharmacodynamic evaluation of the protective effect of compound 43 on kidney injury caused by bacterial infection

[0360] Use creatinine (CRE) measurement and urea nitrogen (BUN) measurement (diacetyl monoxime colorimetric method) to detect the liver function of mice.

[0361] CRE Determination

[0362] The CRE assay kit was purchased from Nanjing Jiancheng Bioengineering Research Institute Co., Ltd., product number: C011-2-1.

[0363] Add 180 μL of enzyme solution A to each of the assay group (T), standard group (S), and blank group (B). Add 6 μL of the sample to be tested to the assay group, 6 μL of the standard product to the standard group, and 6 μL of double-distilled water to the blank group, and incubate at 37 °C for 5 minutes. Measure the absorbance (OD value) A1 of each group at a wavelength of 546 nm using an enzyme-linked immunosorbent assay (ELISA) reader.

[0364] Then add 60 μL of enzyme solution B to each of the assay group, standard group, and blank group, and incubate at 37 °C for 5 minutes. Measure the absorbance (OD value) A2 of each group at a wavelength of 546 nm using an ELISA reader.

[0365] Calculate ΔA = A2 – K × A1 for each group, where K is the dilution factor with a value of 186 / 246.

[0366] According to the calculated ΔA values of the assay group, standard group, and blank group (denoted as ΔA T 、ΔA S 、ΔA B ), calculate the creatinine concentration (in μmol / L): C × (ΔA T –ΔA B ) / (ΔA S –ΔA B ), where C is the standard product concentration, i.e., 442 μmol / L.

[0367] BUN Determination

[0368] The BUN assay kit was purchased from Nanjing Jiancheng Bioengineering Research Institute Co., Ltd., product number: C013-1-1.

[0369] Add 1 mL of 1 g / L oxim solution and 1 mL of acid application solution to each of the assay group (T), standard group (S), and blank group (B). Add 0.02 mL of the sample to be tested to the assay group, 0.02 mL of 10 mmol / L BUN standard product to the standard group, and 0.02 mL of double-distilled water to the blank group, mix well, heat in a water bath at 100 °C for 15 minutes, immediately cool with tap water, and pipette 200 μL from the reaction solution into a 96-well plate (be careful not to aspirate air bubbles). Measure the absorbance (OD value) A of each group at a wavelength of 520 nm using an ELISA reader.

[0370] According to the calculated A values of the assay group, standard group, and blank group (denoted as A T 、A S 、A B) Calculate the urea nitrogen concentration (in mmol / L): C×(A T –A B ) / (A S –A B ), where C is the standard concentration, i.e., 10 mmol / L.

[0371] Results

[0372] The CRE measurement results are as shown in (c) of Figure 17 ; the BUN measurement results are as shown in (d) of Figure 17 . It can be seen that the compound of the present application can effectively reduce the CRE level and BUN level of kidney injury caused by bacterial infection, and has a certain protective effect on kidney injury caused by bacterial infection.

[0373] Drug Toxicity Evaluation of Compound 43 in Experimental Example 4

[0374] Grouping of Experimental Animals

[0375] Select 30 male C57 / 6J mice at 7 weeks old, with each mouse weighing about 20 g. Randomly divide them into six groups (blank control group, vehicle control group, compound 43 group, compound 31 group), with 10 mice in each group.

[0376] - Blank control group: Administer normal saline during dosing.

[0377] - Vehicle control group: Administer a mixture of 10% (v / v) DMSO and 90% (v / v) corn oil (vehicle) during dosing.

[0378] - 1 mg / kg group: Administer the vehicle solution of compound 43 during dosing, with a dose of 1 mg of compound 43 per kg of mouse body weight.

[0379] - 25 mg / kg group: Administer the vehicle solution of compound 43 during dosing, with a dose of 25 mg of compound 43 per kg of mouse body weight.

[0380] - 50 mg / kg group: Administer the vehicle solution of compound 43 during dosing, with a dose of 50 mg of compound 43 per kg of mouse body weight.

[0381] - 100 mg / kg group: Administer the vehicle solution of compound 43 during dosing, with a dose of 100 mg of compound 43 per kg of mouse body weight.

[0382] Drug Preparation

[0383] - Vehicle control group: Take 200 μl of DMSO and add 1.8 mL of corn oil, then vortex and mix evenly to obtain the vehicle.

[0384] - 1 mg / kg group: 0.2 mg of Compound 43 was dissolved in 200 μl of DMSO, and corn oil was added to make up to 2 mL. The mixture was vortexed to obtain Solution A.

[0385] - 25 mg / kg group: 5 mg of Compound 43 was dissolved in 200 μl of DMSO, and corn oil was added to make up to 2 mL. The mixture was vortexed to obtain Solution B.

[0386] - 50 mg / kg group: 10 mg of Compound 43 was dissolved in 200 μl of DMSO, and corn oil was added to make up to 2 mL. The mixture was vortexed to obtain Solution C.

[0387] - 100 mg / kg group: 20 mg of Compound 43 was dissolved in 200 μl of DMSO, and corn oil was added to make up to 2 mL. The mixture was vortexed to obtain Solution D.

[0388] Drug Administration

[0389] - Blank control group: 200 μl of normal saline was injected intraperitoneally.

[0390] - Solvent control group: 200 μl of solvent was injected intraperitoneally.

[0391] - 1 mg / kg group: 200 μl of the prepared Solution A was injected intraperitoneally.

[0392] - 25 mg / kg group: 200 μl of the prepared Solution B was injected intraperitoneally.

[0393] - 50 mg / kg group: 200 μl of the prepared Solution C was injected intraperitoneally.

[0394] - 100 mg / kg group: 200 μl of the prepared Solution D was injected intraperitoneally.

[0395] Animal Model Establishment

[0396] 12 hours after the mice were administered the drugs, the mice were intraperitoneally injected with 10% chloral hydrate (20 mg / kg) to anesthetize the mice. One eyeball of the mice was removed with ophthalmic forceps, and the blood was collected into a coagulation-promoting tube and allowed to stand at room temperature for 4 hours. Then, it was centrifuged at 3000 rpm for 10 minutes, and the supernatant was carefully aspirated into a sterile 1.5 mL centrifuge tube and stored in a -80 °C refrigerator as the test sample for Experimental Examples 4.1 - 4.3.

[0397] 4.1 Effects of different doses of Compound 43 on the liver function of mice

[0398] AST determination and ALT determination were carried out, and the specific steps were the same as those described in Experimental Example 3.1.

[0399] The results of AST determination are as shown in Figure 18 (a); The results of ALT determination are as shown in Figure 18As shown in (b). It can be seen that compounds 43 at different doses such as 1 mg / kg, 25 mg / kg, 50 mg / kg, and 100 mg / kg have no obvious effect on the liver function of mice.

[0400] 4.2 Effects of different doses of compound 43 on the renal function of mice

[0401] CRE determination and BUN determination were carried out, and the specific steps were the same as those described in Experimental Example 3.2.

[0402] The results of CRE determination are as shown in Figure 18 (c); The results of BUN determination are as shown in Figure 18 (d). It can be seen that compounds 43 at different doses such as 1 mg / kg, 25 mg / kg, 50 mg / kg, and 100 mg / kg have no obvious effect on the renal function of mice.

[0403] 4.3 Effects of different doses of compound 43 on the cardiac function of mice

[0404] The cardiac function of mice was detected by using the determination of creatine kinase MB isoenzyme (CK-MB) and cardiac troponin I (cTn-I).

[0405] Determination of Creatine Kinase MB Isoenzyme (CK-MB) in the Heart

[0406] Add 50 μL of standard product diluent to the zero well, add 50 μL of standard products at each concentration to each standard well, add 50 μL of the sample to be tested to the sample well, and then add 50 μL of biotin antigen working solution to the zero well, standard wells, and sample wells. Incubate at 37 °C for 30 minutes.

[0407] Then wash, and add 50 μL of avidin-HRP working solution to the zero well, each standard well, and the sample well, and incubate at 37 °C for 30 minutes.

[0408] Then wash, first add 50 μL of chromogenic agent A and then add 50 μL of chromogenic agent B to the blank well, zero well, each standard well, and the sample well, and develop color at 37 °C in the dark for about 10 minutes.

[0409] When obvious gradients appear in several standard wells, add 50 μL of stop solution to the blank well, zero well, each standard well, and the sample well to terminate the reaction.

[0410] Measure the absorbance (OD value) of each well at a wavelength of 450 nm with an enzyme-labeled instrument, and calculate the CK-MB concentration in the sample well.

[0411] Determination of Cardiac Troponin I (cTn-I)

[0412] Add 50 μL of standard diluent to the zero well, add 50 μL of standards at each concentration to each standard well, add 50 μL of the sample to be tested to the sample well, and then add 50 μL of biotinylated antigen working solution to the zero well, standard wells, and sample well. Incubate at 37 °C for 30 minutes.

[0413] Then wash, add 50 μL of avidin-HRP working solution to the zero well, each standard well, and the sample well, and incubate at 37 °C for 30 minutes.

[0414] Then wash, add 50 μL of chromogen A to the blank well, zero well, each standard well, and sample well first, then add 50 μL of chromogen B, and develop color at 37 °C in the dark for about 10 minutes.

[0415] When an obvious gradient appears in several standard wells, add 50 μL of stop solution to the blank well, zero well, each standard well, and sample well to terminate the reaction.

[0416] Measure the absorbance (OD value) of each well at a wavelength of 450 nm with an enzyme-linked immunosorbent assay (ELISA) reader, and calculate the cTn-I concentration in the sample well.

[0417] Results

[0418] The CK-MB determination results are as shown in Figure 18 (e); the cTn-I determination results are as shown in Figure 18 (f). It can be seen that compound 43 at different doses such as 1 mg / kg, 25 mg / kg, 50 mg / kg, and 100 mg / kg has no obvious effect on the cardiac function of mice.

[0419] 4.4 Effects of different doses of compound 43 on the proliferation of Caco-2 cells and CY h RAW cells

[0420] Use the MTT assay to detect the effects of different doses of compound 43 on the proliferation of Caco-2 cells and CY h RAW cells.

[0421] Seed 5000 to 10000 cells per well into a 96-well plate and allow them to adhere for 12 hours. Aspirate the supernatant medium, wash once with PBS, add 100 μL of medium with different concentrations of compound 43 to each well. The final concentrations of compound 43 in each group are 0, 80 μM, 40 μM, 20 μM, 10 μM, 5 μM, 0.5 μM, 50 nM, and 5 nM, and stimulate for 12 hours. Add 10 μL of MTT solution to each well to make the final concentration of MTT 0.5 mg / mL. After culturing for 4 h, carefully aspirate all the supernatant in the wells to prevent the monolayer cells from rupturing. Add 100 μL of DMSO to dissolve the formazan in the cells Crystallize and measure the absorbance at 570 nm using a microplate reader.

[0422] The results are as Figure 19 shown in (a) and (b) of h Compound 43 at different doses had no significant effect on the proliferation of Caco-2 cells and CY

[0423] The above embodiments are only the preferred embodiments of the present application, and the scope of protection of the present application cannot be limited thereby. Any non-substantial changes and substitutions made by those skilled in the art on the basis of the present application fall within the scope of protection required by the present application.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, (I)， in, R 1 for Fluorophenyl, trifluorophenyl, or ethynylphenyl; or Methyl is optionally monosubstituted with pyridyl, pyrrolyl, quinolyl, indolyl, or thienyl.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that R 1 It is fluorophenyl or ethynylphenyl.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R 1 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2,4,5-trifluorophenyl, 2-ethynylphenyl, 3-ethynylphenyl, 4-ethynylphenyl, 6-methylpyridin-3-yl, 1-methyl-1- H -pyrrol-2-yl, 1-methyl-1 H -indol-3-yl, quinolin-2-yl, quinolin-3-yl, thiophen-2-yl, or thiophen-3-yl.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R 1 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2,4,5-trifluorophenyl, 2-ethynylphenyl, 3-ethynylphenyl, 4-ethynylphenyl, 6-methylpyridin-3-yl, 1-methyl-1- H -pyrrol-2-yl, 1-methyl-1 H -indol-3-yl, quinolin-2-yl, or thiophen-3-yl.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R 1 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2,4,5-trifluorophenyl, 2-ethynylphenyl, 3-ethynylphenyl, 4-ethynylphenyl, 6-methylpyridin-3-yl, 1-methyl-1- H -pyrrol-2-yl, or 1-methyl-1 H -indol-3-yl.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R 1 It is 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2,4,5-trifluorophenyl, 2-ethynylphenyl, 3-ethynylphenyl, 4-ethynylphenyl.

7. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R 1 is 2-ethynylphenyl or 1-methyl-1 H -indol-3-yl.

8. A method for preparing a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, characterized in that: include: 5-(2-(triphenyl-λ 5 -phosphine)acetyl)-1,3-dimethyl-1,3-dihydro-2 H -Benzo[ d ] imidazol-2-one and R 1 -CHO reaction to obtain the compound of formula (I) or a pharmaceutically acceptable salt thereof.

9. The preparation method according to claim 8, characterized in that: Also includes: 5-(2-chloroacetyl)-1,3-dimethyl-1,3-dihydro-2 H -Benzo[ d ] imidazol-2-one and triphenylphosphine were reacted to obtain 5-(2-(triphenyl-λ 5 -phosphine)acetyl)-1,3-dimethyl-1,3-dihydro-2 H -Benzo[ d ]Imidazol-2-one.

10. The preparation method according to claim 9, characterized in that: Also includes: 1,3-dimethyl-1,3-dihydro-2 H -Benzo[ d ] imidazol-2-one and chloroacetyl chloride were reacted to obtain 5-(2-chloroacetyl)-1,3-dimethyl-1,3-dihydro-2 H -Benzo[ d ]Imidazol-2-one.

11. Use of the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof in the preparation of a CYP1A1 inhibitor.

12. The use according to claim 11, characterized in that The CYP1A1 inhibitor is a drug for treating or preventing a disease or condition; Wherein, the disease or condition is tumor, bacterial infection, sepsis, inflammation, or organ damage.

13. The use according to claim 12, characterized in that The disease or disorder is bacterial infection or sepsis, inflammation, or organ damage caused by the infection.

14. The use according to claim 12 or 13, characterized in that The bacteria are Staphylococcus aureus, Streptococcus pneumoniae, Enterococcus faecalis, Enterococcus faecium, Enterococcus avium, Mycobacterium tuberculosis, Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Mycobacterium tuberculosis, Morganella morganii, Providencia stuartii, or Pseudomonas aeruginosa.

15. The use according to claim 12 or 13, characterized in that The bacteria is Staphylococcus aureus or Acinetobacter baumannii.

16. The use according to claim 12 or 13, characterized in that The inflammation and / or organ damage is inflammation or damage of one or more of the skin, muscles, bones, brain, cerebellum, brain stem, spinal cord, eyes, ears, nose, tongue, heart, blood vessels, throat, trachea, lungs, esophagus, stomach, small intestine, large intestine, liver, gallbladder, pancreas, kidney, bladder, urethra, ovary, uterus, testicles, spleen, thymus, lymphatic vessels, or lymph nodes.

17. The use according to claim 12 or 13, characterized in that The inflammation and / or organ damage is inflammation or damage of one or more of the liver, kidney, or heart.

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