Thyroid hormone receptor beta selective agonist compounds, pharmaceutical compositions thereof and uses thereof

Abstract

The present invention relates to thyroid hormone receptor β-selective agonist compounds represented by general formula I, their pharmaceutical compositions and uses. The compounds have improved selectivity and drug formability for THR-α while maintaining good THR-β agonist activity, and have shown some activity in in vivo pharmacodynamic assays. JPEG2023553173000142.jpg33159

Description

[Technical Field]

[0001] The present invention relates to agonists selective for thyroid hormone receptor β. More specifically, the present invention relates to thyroid hormone receptor β subtype agonist compounds represented by general formula (I), pharmaceutical compositions thereof, and their use in the manufacture of medicaments for treating related diseases. [Background technology]

[0002] Thyroid hormones (THs) are produced in the thyroid gland and secreted into the circulation (hypothalamus-pituitary-thyroid axis) as two distinct forms: 3,5,3',5'-tetraiodo-L-thyronine (T4) and 3,5,3'-triiodo-L-thyronine (T3). While T4 is the predominant form secreted by the thyroid gland, T3 is the more physiologically active form. T4 is present in all tissues but is converted to T3 by tissue-specific deiodinases present primarily in the liver and kidney. The biological activity of thyroid hormones is mediated by thyroid hormone receptors (THRs). THRs are encoded by distinct genes, expressing α and β receptors located on chromosomes 17 and 3, respectively. Alternative shearing of the primary transcript generates distinct protein subtypes, THRα1, THRα2, THRβ1, and THRβ2, respectively. THRβ1, THRβ1, and THRβ2 are derived from differential promoter expression, and these two subtypes differ only in their amino termini. THRα1 and THRα2 are derived from differential splicing of precursor mRNA and differ primarily in their carboxyl termini. Of these, THRα1, THRβ1, and THRβ2 can bind thyroid hormone. Thyroid hormone receptor subtypes have been shown to differentially contribute to specific physiological responses. THRβ1 plays an important role in regulating thyroid-stimulating hormone in the liver. THRβ2 plays a major role in regulating thyroid-stimulating hormone. Thyroid hormones play a role in lowering serum low-density lipoprotein (LDL) levels. Hyperthyroidism is associated with decreased serum total cholesterol, which is due to thyroid hormones increasing hepatic LDL receptor expression and promoting cholesterol metabolism into bile acids. Hypothyroidism is associated with hypercholesterolemia, and thyroid hormone replacement therapy is known to lower total cholesterol. Thyroid hormones may also reduce the risk of atherosclerosis and other cardiovascular diseases. The incidence of atherosclerotic vascular disease is directly related to LDL cholesterol levels.Thyroid hormones may reduce body weight, improve obesity-associated comorbidities, and have beneficial effects on glycemic control in obese patients with type 2 diabetes by increasing metabolic rate, oxygen consumption, and heat production.

[0003] The development of thyroid mimetics that maintain the beneficial effects of thyroid hormones while avoiding the side effects of hyperthyroidism and hypothyroidism could open new therapeutic avenues for patients with metabolic disorders such as obesity, hyperlipidemia, hypercholesterolemia, and diabetes, as well as hepatic steatosis, non-alcoholic steatohepatitis, atherosclerosis, cardiovascular disease, hypothyroidism, thyroid cancer, thyroid disorders, and related conditions and diseases.

[0004] MGL-3196 is a first-in-class, orally available, small-molecule, hepatic thyroid hormone receptor β (THR-β) selective agonist. Preclinical toxicology and Phase 1 clinical data suggest that MGL-3196 significantly reduced LDL cholesterol, triglycerides, and lipoproteins for the treatment of nonalcoholic steatohepatitis (NASH) and dyslipidemia (Journal of Hepatology, 2018, vol. 68, s37-s64). This suggests that MGL-3196 may be an ideal candidate for reducing cardiovascular risk in patients with NASH, as well as in patients with dyslipidemia receiving moderate doses of statins or who are intolerant to statins (European Heart Journal, Volume 39, Issue suppl_l, August 2018, ehy566.P5387). Phase 2 clinical data showed that adverse events (AEs) were primarily mild (85%) and moderate (15%), with three cases of serious AEs unrelated to treatment (Journal of Hepatology, 2018, vol. 68, S37_S64). [ka]

[0005] Although MGL-3196 is effective as a THR-β agonist in treating various diseases, it remains difficult to discover new compounds that have the beneficial effects of thyroid hormones while avoiding side effects and that have good oral bioavailability and drug formation properties. Therefore, there remains a need in the art for the development of THR-β selective agonists with better specificity, pharmacodynamics, and pharmacokinetic properties. The present invention provides such compounds. Summary of the Invention

[0006] One object of the present invention is to provide a compound represented by general formula (I), a pharmacologically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, crystalline polymorph, solvate, or isotope-labeled compound (including deuterium substitution) thereof.

[0007] Another object of the present invention is to provide a method for preparing said compound.

[0008] Another object of the present invention is to provide a pharmaceutical composition comprising said compound.

[0009] Another object of the present invention is to provide the use of said compounds in the manufacture of a medicament.

[0010] According to one aspect of the present invention, there is provided a compound represented by general formula (I), a pharmacologically acceptable salt thereof, a stereoisomer, an enantiomer, a diastereomer, an atropisomer, a racemate, a crystalline polymorph, a solvate, or an isotope-labeled compound thereof. [ka]

[0011] where: [ka] [ka]

[0012] R0 is hydrogen and C 1-10 alkyl groups;

[0013] R1 is hydrogen, substituted or unsubstituted C 1-10 Alkyl groups, substituted or unsubstituted C 3-10 Cycloalkyl groups, substituted or unsubstituted 3- to 10-membered heterocycloalkyl groups, substituted or unsubstituted C 6-10 and a substituted or unsubstituted 5- to 10-membered heteroaryl group, wherein the substituent is a halogen atom, a hydroxy group, ═O, C 1-6 Alkoxy group, C 1-6 Alkyl group, C 3-10 Cycloalkyl groups, C 6-10 Aryl group, haloC 6-10 Aryl group, C 1-10 Alkyl group C 6-10 Aryl group, C 1-10 Alkoxy group C 6-10 Aryl groups, 5-10 membered heteroaryl groups, C 1-10 alkyl groups, 5-10 membered heteroaryl groups, halo 5-10 membered heteroaryl groups, 3-10 membered heterocycloalkyl groups, and -NR 10 R 11 Selected from the group consisting of:

[0014] R2, R3 and Y are each independently hydrogen, a halogen atom, or a substituted or unsubstituted C 1-6 Alkyl groups, substituted or unsubstituted C 3-6 Cycloalkyl groups, substituted or unsubstituted C 1-6 alkoxy groups, and the substituents of the substitution are selected from the group consisting of halogen atoms, hydroxy groups, C 1-6 Alkyl groups and C 1-6 alkoxy groups;

[0015] R4 is hydrogen, cyano group, -NR 10 R 11 , substituted or unsubstituted C 1-6 Alkyl groups, substituted or unsubstituted C 3-6Cycloalkyl groups and substituted or unsubstituted C 2-8 alkynyl groups, and the substituents of the substituted groups are selected from the group consisting of halogen atoms, hydroxy groups, cyano groups, and C 1-6 alkoxy groups;

[0016] R5 is hydrogen, substituted or unsubstituted C 1-6 Alkyl groups and substituted or unsubstituted C 3-6 cycloalkyl groups, wherein the substituents are selected from the group consisting of halogen atoms, hydroxy groups, and C 1-6 alkoxy groups;

[0017] R6 is hydrogen and substituted or unsubstituted C 1-6 alkyl groups, and the substituents of the substitution are selected from the group consisting of halogen atoms, hydroxy groups and C 1-6 alkoxy groups;

[0018] L is absent or -NR 10 C(O)- or -NR 10 CR 11 R 11 - and;

[0019] Each R 10 are independently hydrogen and substituted or unsubstituted C 1-3 alkyl groups, and the substituents of the substitution are selected from the group consisting of halogen atoms, hydroxy groups and C 1-3 alkoxy groups;

[0020] Each R 11 are independently hydrogen and substituted or unsubstituted C 1-6 alkyl groups, and the substituents of the substitution are selected from the group consisting of halogen atoms, hydroxy groups and C 1-6 alkoxy groups;

[0021] X1 and X2 are independently N and CR 12 R 12is hydrogen, halogen, cyano group, -NR b R c , -C(=O)R a , -C(=O)OR b , -C(=O)NR b R c , substituted or unsubstituted C 1-4 Alkyl groups, substituted or unsubstituted C 3-6 Cycloalkyl groups, substituted or unsubstituted C 6-10 and a substituted or unsubstituted 5- to 10-membered heteroaryl group, wherein the substituents are selected from the group consisting of a halogen atom, a hydroxy group, and C 1-6 alkoxy groups;

[0022] X3 and X4 are independently N and CR 13 R 13 is hydrogen, halogen, cyano group, hydroxy group, -OR a , -NR b R c , -C(=O)R a , -C(=O)OR b , -C(=O)NR b R c , substituted or unsubstituted C 1-4 Alkyl groups and substituted or unsubstituted C 3-6 cycloalkyl groups, wherein the substituents are selected from the group consisting of halogen atoms, hydroxy groups, and C 1-6 alkoxy groups;

[0023] Each R a But independently, C 1-6 Alkyl group, C 3-10 a cycloalkyl group or a 3- to 10-membered heterocycloalkyl group, wherein the alkyl group, cycloalkyl group, and heterocycloalkyl group are independently selected from the group consisting of halogen, hydroxyl group, amino group, and C 1-6 may be substituted with one or more substituents selected from the group consisting of alkyl groups;

[0024] Each R b and R care independently hydrogen, C 1-6 Alkyl group, C 3-10 a cycloalkyl group or a 3- to 10-membered heterocycloalkyl group, wherein the alkyl group, cycloalkyl group, and heterocycloalkyl group are independently selected from the group consisting of halogen, hydroxyl group, amino group, and C 1-6 may be substituted with one or more substituents selected from the group consisting of alkyl groups;

[0025] Alternatively, R b and R c together with the nitrogen atom to which they are attached form a 3-10 membered heterocycloalkyl group, and the heterocycloalkyl group is selected from the group consisting of halogen, hydroxyl, amino and C 1-6 may be substituted with one or more substituents selected from the group consisting of alkyl groups;

[0026] Each occurrence of n is independently 1, 2, or 3.

[0027] According to another aspect of the present invention, there is provided a method for preparing the compounds of the present invention, which method is represented by any of the following schemes:

[0028] Scheme 1: [ka]

[0029] The compound of general formula Ia is dissolved in a polar solvent, and a base is added to react with the compound of general formula Ib to obtain the compound of general formula Ic. The base under the conditions includes inorganic bases (such as sodium carbonate, potassium carbonate, cesium carbonate, lithium hydroxide, sodium hydroxide, or potassium hydroxide) and organic bases (such as triethylamine, N,N-diisopropylethylamine, or pyridine), preferably potassium carbonate. The polar solvent under the conditions includes N,N-dimethylacetamide, N,N-dimethylformamide, dimethyl sulfoxide, or acetonitrile, preferably N,N-dimethylformamide.

[0030] The compound of general formula Ic reacts with a reducing agent to obtain the compound of general formula Id, where the reducing agent under the conditions includes iron powder, sulfide, tin(I) chloride, zinc powder, etc., and the reducing agent is preferably tin(I) chloride.

[0031] The general formula compound Id reacts with a nitrite ester in an acidic acetonitrile solution, followed by further reaction with compound Ie and ring closure at elevated temperature to obtain the general formula compound I-S1. The acid under the conditions is an organic acid, such as a carboxylic acid or a sulfonic acid, preferably acetic acid. The nitrite under the conditions is isoamyl nitrite or tert-butyl nitrite, preferably tert-butyl nitrite, and the A ring, Y, R 2 , R 3 and R 4 is as defined here, or

[0032] Scheme 2: [ka]

[0033] The compound of general formula Id reacts with a nitrite ester under acidic conditions to generate a diazonium salt compound and incorporate a halogen ion to obtain the compound of general formula If; the resulting compound of general formula If is coupled with intermediate Ig under a transition metal catalyst to obtain the compound of general formula I-S1. The acid under these conditions is an organic acid, such as an carboxylic acid or a sulfonic acid, and is preferably acetic acid. The nitrite under these conditions is such as isoamyl nitrite or tert-butyl nitrite, and is preferably tert-butyl nitrite. X is a halogen, and the A ring, Y, and R are preferably substituted or unsubstituted. 2 , R 3 and R 4 is as defined here, or

[0034] Scheme Three: [ka]

[0035] The compound of general formula II-a can be condensed with the compound of general formula Id under basic conditions to form an amide bond with a condensing agent to obtain the compound of general formula II. The base under these conditions includes organic bases such as triethylamine or N,N-diisopropylethylamine, and the condensing agent under these conditions includes carbodiimide, phosphonium, or uronium condensing agents. The A ring, Y, and R 2 and R 3 is as defined here, or

[0036] Scheme Four: [ka]

[0037] The compound of general formula Id reacts with the acid chloride compound of general formula II-b under basic conditions to form an amide bond, thereby obtaining the compound of general formula II. The base under such conditions includes an organic base such as triethylamine or N,N-diisopropylethylamine, and the base is preferably triethylamine. The A ring, Y, R 2 and R 3 is as defined here, or

[0038] Scheme Five: [ka]

[0039] Compound III-a of general formula (Ia) is substituted with compound III-b of general formula (Ia) under basic conditions to give compound III-b. The methyl ether protection on the oxygen atom is then removed under standard conditions, followed by a substitution reaction with a halogenated hydrocarbon having an R group under basic conditions to give intermediate III-d of general formula (Ia). The base used in this reaction can be an inorganic base such as potassium carbonate, cesium carbonate, or sodium hydroxide, or an organic base such as triethylamine or N,N-diisopropylethylamine. The reaction temperature is 50-150°C, and the reaction can be carried out under heating or microwave conditions. The organic solvent can be, but is not limited to, dioxane, DMF, DMSO, tetrahydrofuran, NMP, etc. R, R, R, X, X, X, and X are as defined herein.

[0040] III-d is then reduced by the method of Scheme 1 to give the key intermediate III-e, which then reacts with a nitrite ester, then with compound Ie, and undergoes ring closure to give compound III-f of the general formula.

[0041] According to another aspect of the present invention, there is provided a pharmaceutical composition comprising a therapeutically effective amount of one or more compounds selected from the group consisting of the above-mentioned compound, pharmacologically acceptable salts, stereoisomers, enantiomers, diastereomers, atropisomers, racemates, crystalline polymorphs, solvates, and isotopically labeled compounds (including deuterium substitutions) thereof, and any pharmacologically acceptable additive.

[0042] According to another aspect of the present invention, there is provided use of the above-mentioned compound, a pharmacologically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, crystalline polymorph, solvate or isotopically labeled compound (including deuterium substitution) thereof, or the above-mentioned composition in the manufacture of a medicament for treating a metabolic-related disease.

[0043] According to another aspect of the present invention, there is provided a method for treating a metabolic-related disease, the method comprising administering to a subject an effective amount of one or more selected from the group consisting of the above-described compounds, their pharmacologically acceptable salts, stereoisomers, enantiomers, diastereomers, atropisomers, racemates, crystalline polymorphs, solvates, and isotopically labeled compounds (including deuterium substitutions), or a pharmaceutical composition comprising, as an active ingredient, one or more selected from the group consisting of the above-described compounds, their pharmacologically acceptable salts, stereoisomers, enantiomers, diastereomers, atropisomers, racemates, crystalline polymorphs, solvates, and isotopically labeled compounds (including deuterium substitutions).

[0044] beneficial effects

[0045] The present disclosure has effectively developed THR-β selective agonists with improved specificity, pharmacodynamics, and pharmacokinetic properties that show potential for the treatment of a wide range of diseases. DETAILED DESCRIPTION OF THE INVENTION

[0046] In order to allow those having ordinary skill in the art to understand the features and advantages of the present invention, the following are general explanations and definitions of the terms and expressions referred to in the present specification and patent application. Unless otherwise specified, all technical and scientific terms used herein shall have the ordinary meanings understood by those skilled in the art in relation to the present invention, and in the event of any discrepancy, the definitions in this specification shall prevail.

[0047] As used herein, the terms "comprise," "include," "have," "contain," and other similar terms are open-ended transitional phrases and are intended to cover a non-exclusive inclusion. For example, a composition or product containing multiple elements is not limited to the elements described herein and may include other elements not expressly listed but normally inherent in the composition or product. Furthermore, unless expressly stated to the contrary, the term "or" refers to an inclusive "or," not an exclusive "or." For example, a condition "A or B" is satisfied when A is true (or present) and B is false (or absent), when A is false (or absent) and B is true (or present), or when both A and B are true (or present). Furthermore, for purposes herein, the terms "comprise," "include," "have," and "contain" should be considered to cover both the closed and semi-closed conjunctions "consisting of" and "consisting essentially of," whose interpretations have already been specifically disclosed.

[0048] All characteristics and conditions defined herein in the form of numerical ranges or percentage ranges are intended for brevity and convenience. Accordingly, the description of a numerical range or percentage range shall be deemed to encompass and specifically disclose all possible subranges and individual values, particularly integer values, within that range. For example, description of a range "1 to 8" shall be deemed to specifically disclose all subranges, such as 1 to 7, 2 to 8, 2 to 6, 3 to 6, 4 to 8, 3 to 8, etc., particularly subranges defined by all integer values, and shall be deemed to specifically disclose individual values ​​within the range, such as 1, 2, 3, 4, 5, 6, 7, 8, etc. Unless otherwise specified for individual values, the foregoing interpretation applies to the entire invention, regardless of the breadth of the range.

[0049] When a quantity or other numerical value or parameter is expressed as a range, a more preferred range, or a series of upper and lower limits, all ranges consisting of any pair of the upper or more preferred value and the lower or more preferred value, whether or not such ranges are individually disclosed, are to be understood as specifically disclosed herein. Further, when a range of values ​​is referred to herein, unless otherwise specified, the range is intended to include its endpoints, and all integers and fractions within the range.

[0050] Here, numerical values ​​should be understood to have the precision of the significant digits of that numerical value, provided that the purpose of the present invention is achieved. For example, the numerical value 40.0 should be understood to cover the range from 39.50 to 40.49.

[0051] When a Markush group or a selection term is used to describe a feature or embodiment of the present invention, those skilled in the art will understand that any subgroup of all elements in the Markush group or selection term, or any individual element, can also be used to describe the present invention. For example, when X is described as "selected from the group consisting of X1, X2, and X3," this means that both the assertion that X is X1 and the assertion that X is X1 and / or X2 are fully described. Furthermore, when a Markush group or a selection term is used to describe a feature or embodiment of the present invention, those skilled in the art will understand that any subgroup of all elements in the Markush group or selection list, or any combination of individual elements, can also be used to describe the present invention. Thus, for example, when X is described as "selected from the group consisting of X1, X2, and X3" and Y is described as "selected from the group consisting of Y1, Y2, and Y3," the assertion that X is X1, X2, or X3, and Y is Y1, Y2, or Y3 is fully described.

[0052] The following specific embodiments are exemplary in nature and are not intended to limit the invention and its uses. Furthermore, the present specification is not limited by the above-mentioned prior art or invention, or the theory presented in the following specific embodiments or examples.

[0053] According to one embodiment of the present disclosure, there is provided a compound represented by general formula (I), a pharmacologically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, crystalline polymorph, solvate, or isotopically labeled compound thereof. [ka]

[0054] where: [ka] [ka]

[0055] R0 is hydrogen and C 1-10 alkyl groups;

[0056] R1 is hydrogen, substituted or unsubstituted C 1-10 Alkyl groups, substituted or unsubstituted C 3-10 Cycloalkyl groups, substituted or unsubstituted 3- to 10-membered heterocycloalkyl groups, substituted or unsubstituted C 6-10 and a substituted or unsubstituted 5- to 10-membered heteroaryl group, wherein the substituent is a halogen atom, a hydroxy group, ═O, C 1-6 Alkoxy group, C 1-6 Alkyl group, C 3-10 Cycloalkyl groups, C 6-10 Aryl group, haloC 6-10 Aryl group, C 1-10 Alkyl group C 6-10 Aryl group, C 1-10 Alkoxy group C 6-10Aryl groups, 5-10 membered heteroaryl groups, C 1-10 alkyl groups, 5-10 membered heteroaryl groups, halo 5-10 membered heteroaryl groups, 3-10 membered heterocycloalkyl groups, and -NR 10 R 11 Selected from the group consisting of:

[0057] R2, R3 and Y are each independently hydrogen, a halogen atom, or a substituted or unsubstituted C 1-6 Alkyl groups, substituted or unsubstituted C 3-6 Cycloalkyl groups, substituted or unsubstituted C 1-6 alkoxy groups, and the substituents of the substitution are selected from the group consisting of halogen atoms, hydroxy groups, C 1-6 Alkyl groups and C 1-6 alkoxy groups;

[0058] R4 is hydrogen, cyano group, -NR 10 R 11 , substituted or unsubstituted C 1-6 Alkyl groups, substituted or unsubstituted C 3-6 Cycloalkyl groups and substituted or unsubstituted C 2-8 alkynyl groups, and the substituents of the substituted groups are selected from the group consisting of halogen atoms, hydroxy groups, cyano groups, and C 1-6 alkoxy groups;

[0059] R5 is hydrogen, substituted or unsubstituted C 1-6 Alkyl groups and substituted or unsubstituted C 3-6 cycloalkyl groups, wherein the substituents are selected from the group consisting of halogen atoms, hydroxy groups, and C 1-6 alkoxy groups;

[0060] R6 is hydrogen and substituted or unsubstituted C 1-6 alkyl groups, and the substituents of the substitution are selected from the group consisting of halogen atoms, hydroxy groups and C 1-6 alkoxy groups;

[0061] L is absent or -NR10 C(O) or -NR 10 CR 11 R 11 and;

[0062] Each R 10 are independently hydrogen and substituted or unsubstituted C 1-3 The substituents of the substituents are selected from the group consisting of alkyl groups, halogen atoms, hydroxy groups and C 1-3 alkoxy groups;

[0063] Each R 11 are independently hydrogen and substituted or unsubstituted C 1-6 alkyl groups, and the substituents of the substitution are selected from the group consisting of halogen atoms, hydroxy groups and C 1-6 alkoxy groups;

[0064] X1 and X2 are independently N and CR 12 R 12 is hydrogen, halogen, cyano group, -NR b R c , -C(=O)R a , -C(=O)OR b , -C(=O)NR b R c , substituted or unsubstituted C 1-4 Alkyl groups, substituted or unsubstituted C 3-6 Cycloalkyl groups, substituted or unsubstituted C 6-10 and a substituted or unsubstituted 5- to 10-membered heteroaryl group, wherein the substituents are selected from the group consisting of a halogen atom, a hydroxy group, and C 1-6 alkoxy groups;

[0065] X3 and X4 are independently N and CR 13 R 13 is hydrogen, halogen, cyano group, hydroxy group, -OR a , -NR b R c , -C(=O)R a , -C(=O)ORb , -C(=O)NR b R c , substituted or unsubstituted C 1-4 Alkyl groups and substituted or unsubstituted C 3-6 cycloalkyl groups, wherein the substituents are selected from the group consisting of halogen atoms, hydroxy groups, and C 1-6 alkoxy groups;

[0066] Each R a But independently, C 1-6 Alkyl group, C 3-10 a cycloalkyl group or a 3- to 10-membered heterocycloalkyl group, wherein the alkyl group, cycloalkyl group, and heterocycloalkyl group are independently selected from the group consisting of halogen, hydroxyl group, amino group, and C 1-6 may be substituted with one or more substituents selected from the group consisting of alkyl groups;

[0067] Each R b and R c are independently hydrogen, C 1-6 Alkyl group, C 3-10 a cycloalkyl group or a 3- to 10-membered heterocycloalkyl group, wherein the alkyl group, cycloalkyl group, and heterocycloalkyl group are independently selected from the group consisting of halogen, hydroxyl group, amino group, and C 1-6 may be substituted with one or more substituents selected from the group consisting of alkyl groups;

[0068] Alternatively, R b and R c together with the nitrogen atom to which they are attached form a 3-10 membered heterocycloalkyl group, and the heterocycloalkyl group is selected from the group consisting of halogen, hydroxyl, amino and C 1-6 may be substituted with one or more substituents selected from the group consisting of alkyl groups;

[0069] Each occurrence of n is independently 1, 2, or 3.

[0070] According to one embodiment of the present disclosure, the compound of general formula (I) is selected from the group consisting of compounds represented by formula (Ia) or (Ib). [ka]

[0071] R0 is hydrogen;

[0072] R1 is hydrogen, substituted or unsubstituted C 1-6 Alkyl groups, substituted or unsubstituted C 3-6 Cycloalkyl groups and substituted or unsubstituted C 3-8 heterocycloalkyl groups, wherein the substituents are selected from the group consisting of halogen atoms, hydroxy groups, ═O, and C 1-6 alkoxy groups;

[0073] R2 and R3 are each independently a halogen atom, a substituted or unsubstituted C 1-6 Alkyl groups and substituted or unsubstituted C 3-6 cycloalkyl groups, and the substituents of the substitution are selected from the group consisting of halogen atoms, hydroxy groups, C 1-4 Alkyl groups and C 1-4 alkoxy groups;

[0074] n, each occurrence, is independently 1 or 2;

[0075] X1 and X2 are independently N and CR 12 R 12 is hydrogen, halogen, cyano group, substituted or unsubstituted C 1-4 Alkyl groups, substituted or unsubstituted C 3-6 cycloalkyl groups, wherein the substituents are selected from the group consisting of halogen atoms, hydroxy groups, and C 1-6 alkoxy groups;

[0076] X3 and X4 are independently N and CR 13 R13 is hydrogen, substituted or unsubstituted C 1-4 Alkyl groups and substituted or unsubstituted C 3-6 cycloalkyl groups, wherein the substituents are selected from the group consisting of halogen atoms, hydroxy groups, and C 1-6 alkoxy groups;

[0077] R5 and B ring are as defined above.

[0078] According to one embodiment of the present disclosure, the compound of general formula (I) is selected from the group consisting of compounds represented by formula (Ic): [ka]

[0079] R2 and R3 are each independently a halogen atom, a substituted or unsubstituted C 1-6 Alkyl groups and substituted or unsubstituted C 3-6 cycloalkyl groups, and the substituents of the substitution are selected from the group consisting of halogen atoms, hydroxy groups, C 1-4 Alkyl groups and C 1-4 alkoxy groups;

[0080] X1 and X2 are independently N and CR 12 R 12 is hydrogen, halogen, cyano group, substituted or unsubstituted C 1-4 Alkyl groups, substituted or unsubstituted C 3-6 cycloalkyl groups, wherein the substituents are selected from the group consisting of halogen atoms, hydroxy groups, and C 1-6 alkoxy groups;

[0081] X3 and X4 are independent of each other, 13 and R 13 is hydrogen, substituted or unsubstituted C 1-4 Alkyl groups and substituted or unsubstituted C 3-6cycloalkyl groups, wherein the substituents are selected from the group consisting of halogen atoms, hydroxy groups, and C 1-6 alkoxy groups;

[0082] R1 and B-ring are as defined above.

[0083] According to one embodiment of the present disclosure, the compound of general formula (I) is selected from the group consisting of compounds represented by formula (Id): [ka]

[0084] R2 and R3 are each independently selected from the group consisting of halogen atoms;

[0085] L is -NHC(O)- or -NHCHR 11 - and;R 11 is hydrogen and substituted or unsubstituted C 1-6 alkyl groups, and the substituents of the substitution are selected from the group consisting of halogen atoms, hydroxy groups and C 1-6 alkoxy groups;

[0086] X1 and X2 are independently N and CR 12 R 12 is hydrogen, halogen, cyano group, substituted or unsubstituted C 1-4 Alkyl groups, substituted or unsubstituted C 3-6 cycloalkyl groups, wherein the substituents are selected from the group consisting of halogen atoms, hydroxy groups, and C 1-6 alkoxy groups;

[0087] R1 is as defined above.

[0088] According to one embodiment of the present disclosure, the compound of general formula (I) is selected from the group consisting of compounds represented by formula (Ie): [ka]

[0089] R2 and R3 are each independently selected from the group consisting of halogen atoms;

[0090] X1 and X2 are independently N and CR 12 R 12 is hydrogen, halogen, cyano group, substituted or unsubstituted C 1-4 Alkyl groups, substituted or unsubstituted C 3-6 cycloalkyl groups, wherein the substituents are selected from the group consisting of halogen atoms, hydroxy groups, and C 1-6 alkoxy groups;

[0091] R1 and R4 are as defined above.

[0092] According to one embodiment of the present disclosure, the compound of general formula (I) is selected from the group consisting of compounds represented by formula (If): [ka]

[0093] R2 and R3 are each independently selected from the group consisting of halogen atoms;

[0094] X1 and X2 are independently N and CR 12 R 12 is hydrogen, halogen, cyano group, substituted or unsubstituted C 1-4 Alkyl groups, substituted or unsubstituted C 3-6 cycloalkyl groups, wherein the substituents are selected from the group consisting of halogen atoms, hydroxy groups, and C 1-6 alkoxy groups;

[0095] R1 and R6 are as defined above.

[0096] In the present invention, the heteroatom in the heterocycloalkyl group or heteroaryl group is one or more selected from the group consisting of O, N, and S, and the sulfur atom may be oxidized to form a sulfoxide group or a sulfone group.

[0097] According to one embodiment of the present disclosure, the compound of general formula I is selected from the group consisting of the following compounds: [ka] JPEG0007729891000019.jpg214159JPEG0007729891000020.jpg215159JPEG0007729891000021.jpg191159JPEG0007729891 000022.jpg231159JPEG0007729891000023.jpg221159JPEG0007729891000024.jpg221159JPEG0007729891000025.jpg58159

[0098] According to one embodiment of the present disclosure, there is provided a pharmaceutical composition comprising one or more selected from the group consisting of the above-mentioned compound, a pharmacologically acceptable salt thereof, a stereoisomer, an enantiomer, a diastereomer, an atropisomer, a racemate, a crystalline polymorph, a solvate, and an isotope-labeled compound thereof, and an optional pharmacologically acceptable additive.

[0099] According to one embodiment of the present disclosure, there is provided use of the above-mentioned compound, a pharmacologically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, crystalline polymorph, solvate or isotopically labeled compound thereof, or the above-mentioned pharmaceutical composition in the manufacture of a medicament for treating a metabolic-related disease.

[0100] According to one embodiment of the present disclosure, there is provided a method for treating a metabolic-related disease, comprising administering to a subject an effective amount of one or more selected from the group consisting of the compound, its pharmacologically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, crystalline polymorph, solvate, and isotopically labeled compound, or the pharmaceutical composition.

[0101] According to one embodiment of the present disclosure, the metabolic-related disease is selected from the group consisting of obesity, hyperlipidemia, hypercholesterolemia, diabetes, non-alcoholic fatty liver disease (NASH), hepatic steatosis, atherosclerosis, hypothyroidism, and thyroid cancer.

[0102] According to one embodiment of the present disclosure, the metabolic-related disease is selected from the group consisting of non-alcoholic fatty liver disease (NASH), hypothyroidism, and thyroid cancer.

[0103] Example

[0104] In the following examples, the optimal reaction conditions and reaction times for each step may vary depending on the specific reactants used and the substituents present in all reactants.Unless otherwise specified, solvents, temperatures, and other reaction conditions can be easily selected by those skilled in the art.Specific steps are provided in the synthesis examples section.The reaction can be further processed in a conventional manner, for example, but not limited to, removing the solvent from the residue and further purifying according to methods commonly known in the art, such as crystallization, distillation, extraction, trituration, and chromatography.Unless otherwise specified, the starting materials and reactants are commercially available or can be prepared by those skilled in the art from commercially available materials using methods described in the chemical literature.

[0105] Routine experimentation, such as appropriate adjustment of reaction conditions, reactants, and order of synthetic pathways, protection of any chemical functional groups that may be incompatible with the reaction conditions, and deprotection at appropriate points in the reaction sequence of the method, is within the scope of the present invention. Suitable protecting groups and methods for protecting and deprotecting different substituents using such suitable protecting groups are well known to those skilled in the art, and examples are found in T. Greene and P. Wuts, Protecting Groups in Chemical Synthesis (3rd Edition), John Wiley & Sons, NY (1999), which is incorporated herein by reference in its entirety. Synthesis of compounds of the present invention can be achieved by methods similar to those described in the above synthetic schemes and specific examples.

[0106] If the starting materials are not commercially available, they can be prepared by processes selected from standard organic chemistry techniques, techniques similar to those used to synthesize known structural analogs, or techniques similar to those described in the above schemes or synthetic examples. When an optically active form of a compound of the present invention is desired, it can be obtained by carrying out one of the processes described herein using an optically active starting material (e.g., prepared by asymmetric induction in an appropriate reaction step), or by resolving a mixture of stereoisomers of the compound or intermediate using standard processes (e.g., chromatographic separation, further crystallization, or enzymatic resolution).

[0107] Similarly, if a pure geometric isomer of a compound of the invention is desired, it can be obtained by performing one of the above processes starting with a pure geometric isomer, or by using standard processes such as chromatographic separation of a mixture of geometric isomers of a resolved compound or intermediate.

[0108] For illustrative purposes, the following examples can be used, and the following embodiments are only intended to illustrate the technical solutions of the present invention, and are not intended to limit the present invention to these examples.

[0109] Example 1: Preparation of compounds ZB-H-01 and ZB-H-02 [ka]

[0110] Step 1: Preparation of Compound 1b

[0111] To a solution of 5-methoxy-1H-indole (1a, 1.47 g, 10 mmol) in N,N-dimethylformamide (20 mL), 1,3-dichloro-2-fluoro-5-nitrobenzene (2.5 g, 12 mmol) and potassium carbonate (2 g, 15 mmol) were added, and the mixture was heated to 100 °C and stirred overnight. The reaction was stopped, cooled to room temperature, and saturated brine (100 mL) was added. The mixture was extracted three times with ethyl acetate. The organic phases were combined and washed with saturated brine. The solvent was evaporated to dryness under reduced pressure to give compound 1b (3 g). This product was used directly in the next step. LC-MS [M+H] + :338.

[0112] Second step: Preparation of compound 1c

[0113] Tin(I) chloride dihydrate (3.4 g, 14.9 mmol) was added to a solution of compound 1b (1 g, 2.98 mmol) in ethanol (20 mL). After the reaction was complete, the mixture was heated to 80°C and stirred for 6 hours. After the reaction was stopped, the mixture was allowed to cool to room temperature, and the solvent was concentrated under reduced pressure until dry. Ethyl acetate (200 mL) was added and dissolved, and the organic phase was washed three times with aqueous sodium hydroxide (2 M). The organic phase was concentrated and then purified by column chromatography to give compound 1c (750 mg). 1 H NMR(400MHz,CDCl3) δ 7.16(s,1H),7.05(d,J=3.2Hz,1H),6.90-6.82(m,2H),6.74(s,2H),6.62(d,J=3.2Hz,1H),3.96(s,2H),3.87(s,3H);LC-MS[M+H] + :307.

[0114] Third step: Preparation of compound ZB-H-01

[0115] Tert-butyl nitrite (202 mg, 1.9 mmol) was dissolved in acetic acid (3 mL). At 0°C, this solution was slowly added to a solution of compound 1c (500 mg, 1.63 mmol) in acetic acid (20 mL) and acetonitrile (10 mL). The mixture was stirred for 30 minutes while maintaining the temperature at 0°C. A solution of N-cyanoacetylurethane (1d, 330 mg, 2.1 mmol) in acetonitrile was then added dropwise. After the addition was complete, the mixture was stirred for 3 hours. After quenching the reaction, the reaction mixture was added to saturated aqueous sodium bicarbonate (150 mL). The resulting red solid was filtered, washed with water and petroleum ether, and dried. The resulting solid was added with N,N-dimethylacetamide (5 ml) and potassium acetate (1.9 mmol), heated to 120 °C and stirred for 6 hours, cooled to room temperature, added with 50 ml of water, extracted three times with ethyl acetate, and the organic phases were combined. The organic phase was washed with saturated brine, and the solvent was concentrated under reduced pressure to dryness. Compound ZB-H-01 (520 mg) was obtained by column chromatography isolation and purification. LC-MS [M+H] + :428.

[0116] Fourth step: Preparation of compound ZB-H-02

[0117] Compound ZB-H-01 (30 mg) was dissolved in 20 mL of DCM and purged with argon three times. A solution of boron tribromide in dichloromethane (2 M, 0.7 mL) was slowly added to the solution at -78 °C, and the temperature was gradually raised to -10 °C. The reaction was monitored by TLC. After the starting material disappeared, 10 mL of saturated aqueous sodium bicarbonate was added, and the mixture was allowed to cool to room temperature while stirring. The mixture was allowed to stand and separated. The dichloromethane layer was separated and extracted three times with ethyl acetate. The organic layers were combined, dried, and the solvent was concentrated under reduced pressure to dryness. Compound ZB-H-02 (18 mg) was obtained by thin-layer chromatography (DCM:MeOH = 8:1). 1H NMR(400MHz,DMSO-d6) δ 8.89(s,1H),7.89(s,2H),7.36(d,J=3.2Hz,1H),6.96(s,1H),6.75(d,J=8.7Hz,1H),6.65(d,J=8.7Hz,1H),6.55(d,J=3.2Hz,1H);LC-MS[M+H] + :414.

[0118] Example 2: Preparation of compound ZB-H-07 [ka]

[0119] Using the synthetic route of Example 1, except that starting material 1a was replaced with starting material 2a in the first step, the other steps and conditions were all the same as in Example 1, and the title product ZB-H-07 was finally produced. 1 H NMR(400MHz,DMSO-d6) δ 9.41(s,1H),8.25(s,1H),7.92(s,2H),7.20-7.05(m,2H),7.03-6.93(m,1H);LC-MS[M+H] + :415.

[0120] Example 3: Preparation of compound ZB-H-08 [ka]

[0121] Using the synthetic route of Example 1, except that starting material 1a was replaced with 3a in the first step, the other steps and conditions were all the same as in Example 1, and the title product ZB-H-08 was finally produced. 1 H NMR(400MHz,DMSO-d6) δ 13.30(s,1H),9.30(s,1H),7.79(s,2H),7.45(d,J=8.6Hz,1H),6.72(s,1H),6 .61(d,J=7.3Hz,1H),4.26(t,J=8.4Hz,2H),3.18(t,J=8.4Hz,2H);LC-MS[M+H] + :416.

[0122] Example 4: Preparation of compound ZB-H-09 [ka]

[0123] Using the synthetic route of Example 1, except that starting material 1a was replaced with 4a in the first step, the other steps and conditions were all the same as in Example 1, and the title product ZB-H-09 was finally produced. 1 H NMR(400MHz,DMSO-d6) δ 8.92(s,1H),7.89(s,2H),7.27(d,J=3.2Hz,1H),6.98(s,1H),6.61(s,1H), 6.51(d,J=3.2Hz,1H),3.26-3.21(m,1H),1.12(d,J=6.9Hz,6H);LC-MS[M+H] + :456.

[0124] Example 5: Preparation of Compound ZB-H-11 [ka]

[0125] Using the synthetic route of Example 1, except that starting material 1a was replaced with 5a in the first step, the other steps and conditions were all the same as in Example 1, and the title product ZB-H-11 was finally produced. 1 LC-MS[M+H] + :415.

[0126] Example 6: Preparation of compound ZB-H-15 [ka]

[0127] Using the synthetic route of Example 1, except that starting material 6a was used instead of starting material 1a in the first step, the other steps and conditions were all the same as in Example 1, and the title product ZB-H-15 was finally produced. 1 H NMR(400MHz,DMSO-d6) δ LC-MS[M+H] + :428.

[0128] Example 7: Preparation of compound ZB-H-16 [ka]

[0129] Using the synthetic route of Example 1, except that starting material 1a was replaced with starting material 7a in the first step, the other steps and conditions were all the same as in Example 1, and the title product ZB-H-16 was finally produced. 1 H NMR(400MHz,DMSO-d6) δ 9.44(s,1H),8.18(s,1H),7.92(s,2H),7.11(s,1H),6.98(s,1H),2.21(s,3H);LC-MS[M+H] + :429.

[0130] Example 8: Preparation of compound ZB-H-17 [ka]

[0131] Using the synthetic route of Example 1, except that starting material 1a was replaced with 8a in the first step, the other steps and conditions were all the same as in Example 1, and the title product ZB-H-17 was finally produced. 1H NMR(400MHz,DMSO-d6) δ 13.28(s,1H),8.66(s,1H),7.36(d,J=3.3Hz,1H),6.71(d,J=8.6Hz,1H),6.63(d,J=3.3Hz,1H),6.58(d,J=8.6Hz,1H),2.32(s,3H);LC-MS:[M+H] + :428.

[0132] Example 9: Preparation of compound ZB-H-18 [ka]

[0133] Using the synthetic route of Example 1, except that starting material 1a was replaced with starting material 9a in the first step, the other steps and conditions were all the same as in Example 1, and the title product ZB-H-18 was finally produced. 1 H NMR(400MHz,DMSO-d6) δ 13.37(s,1H),9.12(s,1H),8.36(s,1H),7.92(s,2H),7.02(d,J=8.8Hz,1H),6.91(d,J=8.8Hz,1H),2.40(s,3H); LC-MS[M+H] + :429.

[0134] Example 10: Preparation of compound ZB-H-19 [ka]

[0135] Using the synthetic route of Example 1, except that starting material 1a was replaced with starting material 10a in the first step, the other steps and conditions were all the same as in Example 1, and the title product ZB-H-19 was finally produced. 1 H NMR(400MHz,DMSO-d6) δ 8.92(s,1H),7.86(s,2H),7.14(s,1H),6.88(s,1H),6.71(d,J=8.7Hz,1H),6.65(d,J=8.7Hz,1H),2.24(s,3H);LC-MS[M+H] + :428.

[0136] Example 11: Preparation of compounds ZB-H-22 and ZB-H-23 [ka]

[0137] Using the synthetic route of Example 1, except that starting material 1a was replaced with starting material 11a in the first step, the other steps and conditions were all the same as in Example 1, and the title products ZB-H-22 and ZB-H-23 were finally produced.

[0138] Compound ZB-H-22: 1 H NMR(400MHz,DMSO-d6) δ 11.71(s,1H),7.89(s,2H),7.41(d,J=3.1Hz,1H),7.30(d,J=9.5Hz,1H),6.28(d,J=3.0Hz,1H),6.06(d,J=9.4Hz,1H); LC-MS[M+H] + :415.

[0139] Compound ZB-H-23:LC-MS[M+H] + :429.

[0140] Example 12: Preparation of compound ZB-H-24 [ka]

[0141] Using the synthetic route of Example 1, except that starting material 12a was used instead of starting material 1a in the first step, the other steps and conditions were all the same as in Example 1, and the title product ZB-H-24 was finally produced. 1 H NMR(400MHz,DMSO-d6) δ 8.90(s,1H),7.86(s,2H),7.11(s,1H),6.96(d,J=2.1Hz,1H),6.71(d,J=8.7Hz, 1H),6.63(d,J=8.7Hz,1H),3.15-3.02(m,1H),1.31(d,J=6.8Hz,6H);LC-MS[M+H] + :456.

[0142] Example 13: Preparation of compound ZB-H-31 [ka]

[0143] Using the synthetic route of Example 1, except that starting material 1a was replaced with starting material 13a in the first step, the other steps and conditions were all the same as in Example 1, and the title product ZB-H-31 was finally produced. 1 H NMR(400MHz,DMSO-d6) δ 13.35(s,1H),9.38(s,1H),7.86(s,2H),7.81(s,1H),7.58(d,J=3.6Hz,1H),7.41(s,1H),6.61(d,J=3.6Hz,1H); LC-MS:[M+H] + :415.

[0144] Example 14: Preparation of Compound 14e [ka]

[0145] First step: Preparation of intermediate 14b

[0146] The synthetic route of the first step in Example 1 was adopted, and 11a was used instead of starting material 1a to prepare intermediate 14b. LC-MS [M+H] + :339.

[0147] Second step: Preparation of intermediate 14c

[0148] Compound 14b (337 mg, 1 mmol) was dissolved in anhydrous acetonitrile (10 mL), stirred under argon protection, and sodium iodide (750 mg, 5 mmol) and trimethylchlorosilane (0.6 mL, 5 mmol) were added, refluxed, and monitored by TLC. After the starting material disappeared, the mixture was cooled to room temperature and quenched by adding methanol (5 mL) while maintaining stirring. The solvent was concentrated under reduced pressure to dryness, and compound 14c (249 mg) was obtained by column chromatography isolation and purification. 1 H NMR(400MHz,DMSO-d6) δ 11.70(s,1H),8.61(s,2H),7.40(d,J=3.2Hz,1H),7.29(d,J=9.5Hz,1H),6.32(d,J=3.1Hz,1H),6.09(d,J=9.5Hz,1H);LC-MS[M+H] + :325.

[0149] Third Step: Preparation of Intermediate 14d

[0150] Compound 14c (323 mg, 1 mmol) was dissolved in 1,4-dioxane (5 mL), placed in a microwave reactor, and methyl iodide (3 mmol) and cesium carbonate (3 mmol) were added. The mixture was microwaved (150 °C, 12 h). After the reaction was completed, the solvent was evaporated to dryness under reduced pressure, and the product was purified by column chromatography to give compound 14d (288 mg). LC-MS: [M+H] + :338.

[0151] Fourth step: Preparation of compound 14e

[0152] Tin(I) chloride dihydrate (3.4 g, 14.9 mmol) was added to a solution of compound 14d (1 g, 2.98 mmol) in ethanol (20 mL). After the reaction was complete, the mixture was heated to 80 °C and stirred for 6 hours. After the reaction was stopped, the mixture was allowed to cool to room temperature. The solvent was concentrated under reduced pressure to dryness, and then ethyl acetate (200 mL) was added and dissolved. The organic phase was washed three times with aqueous sodium hydroxide (2 M). The organic phase was concentrated and then purified by column chromatography to give compound 14e. LC-MS [M+H] + :308.

[0153] Example 15: Preparation of Compound 15e [ka]

[0154] Using the synthetic route of Example 14, compound 15e was prepared by using isopropyl iodide instead of methyl iodide in the third step of the synthesis. LC-MS: [M+H] + :336.

[0155] Example 16: Preparation of Compound 16e [ka]

[0156] First step: To a solution of ethyl cyanoformate (16a, 9 g, 106 mmol) in ethanol (100 mL) and water (80 mL) was added hydroxylamine hydrochloride (11 g, 159 mmol) and sodium carbonate (11 g, 106 mmol). The mixture was stirred at room temperature for 2 hours. The organic solvent was removed in vacuo, and the aqueous layer was extracted with dichloromethane (8 × 100 mL). The combined organic layers were dried over MgSO4, filtered, and concentrated to give compound 16b (5.7 g) as a white solid. LC-MS [M+H] + :133.

[0157] Second step: N,N'-Carbonyldiimidazole (9.4 g, 58 mmol) and 1,8-diazabicycloundecane-7-ene (8.7 g, 58 mmol) were added to a solution of compound 16b (5.7 g, 48 mmol) in 1,4-dioxane (50 mL) and stirred at 80 °C for 2 h. The reaction was quenched with HCl, concentrated, and extracted with dichloromethane (8 × 100 mL). The combined organic layers were concentrated and purified by column chromatography (DCM:EA = 8:1) to give compound 16c (3 g) as a yellow oil. 1 H NMR(400MHz,DMSO-d6) δ 13.41(s,1H),4.37(q,J=7.1Hz,2H),1.30(t,J=7.1Hz,3H);LC-MS[MH] - :157.

[0158] Third step: Lithium hydroxide (0.5 g, 20.8 mmol) was added to the compound 16c The resulting mixture was added to a mixture of 16d (3 g, 20.8 mmol) in tetrahydrofuran (30 mL) and water (30 mL) and stirred at room temperature for 3 hours. The mixture was washed with ethyl acetate (2 × 20 mL) and acidified with 1 M HCl to pH = 3, concentrated, extracted with ethyl acetate (8 × 20 mL), dried, and the solvent was evaporated to dryness under reduced pressure to give compound 16d (2 g) as a white solid. LC-MS [MH] - :129.

[0159] Fourth step: Compound 16d (2 g, 15.4 mmol) was dissolved in anhydrous tetrahydrofuran, and 1 drop of N,N-dimethylformamide was added. The mixture was stirred at 0 °C, and oxalyl chloride (1.3 mL, 15.4 mmol) was added dropwise. The mixture was allowed to warm to room temperature and stirred for 30 min. The solvent was concentrated under reduced pressure to dryness to give compound 16e (1.8 g).

[0160] Example 17: Preparation of compound ZB-H-25 [ka]

[0161] The third step in Example 1 About Adopting this synthetic route, compound 14e was used in place of 1c in the third step to prepare the title compound ZB-H-25. 1 H NMR(400MHz,DMSO-d6) δ 7.90(s,2H),7.51(d,J=3.2Hz,1H),7.32(d,J=9.4Hz,1H),6.61(d,J=3.0Hz,1H),6.15(d,J=9.4Hz,1H),3.55(s,3H); LC-MS:[M+H] + :429.

[0162] Example 18: Preparation of compound ZB-H-26 [ka]

[0163] The third step in Example 1 About Adopting this synthetic route, compound 15e was used in place of 1c in the third step to prepare the title compound ZB-H-26. 1 H NMR(400MHz,DMSO-d6) δ 13.35(s,1H),7.89(s,2H),7.50(d,J=3.2Hz,1H),7.27(d,J=9.4Hz,1H),6.76(d ,J=3.2Hz,1H),6.12(d,J=9.4Hz,1H),5.47-5.06(m,1H),1.51(d,J=7.0Hz,6H); LC-MS[M+H] + :458.

[0164] Example 19: Preparation of compound ZB-H-32 [ka]

[0165] Compound 15e (63 mg, 0.19 mmol) was dissolved in anhydrous tetrahydrofuran (2 mL), triethylamine (0.6 mL, 0.9 mmol) was added, and the mixture was cooled to 0°C with stirring. Then, a solution of intermediate 16e (125 mg, 0.9 mmol) in anhydrous tetrahydrofuran (2 mL) was slowly added dropwise. The mixture was allowed to return to room temperature and stirred for 30 minutes. The solvent was then concentrated under reduced pressure to dryness, and the mixture was purified by column chromatography to give compound ZB-H-32 (48 mg). 1 H NMR(400MHz,DMSO-d6) δ 13.49(s,1H),11.52(s,1H),8.12(s,2H),7.41(d,J=3.2Hz,1H),7.17(d,J=9.3Hz,1H), 6.70(d,J=3.3Hz,1H),6.10(d,J=9.4Hz,1H),5.40-5.17(m,1H),1.50(d,J=7.0Hz,6H); LC-MS[M+H] + :449.

[0166] Example 20: Preparation of Compound 20e [ka]

[0167] Step 1: Preparation of Compound 20d

[0168] Compound 14c (323 mg, 1 mmol) was dissolved in toluene (5 mL) and placed in a microwave reactor. Cyclopropylboronic acid (3 mmol), copper acetate (1 mmol), pyridine (5 mmol), and sodium bis(trimethylsilyl)amide (1 mmol) were added, and the mixture was heated under oxygen gas at 120°C for 3 hours. After the reaction was completed, the solvent was evaporated to dryness under reduced pressure, and compound 20d (300 mg) was obtained by column chromatography purification.

[0169] Second step: Preparation of compound 20e

[0170] The first in Example 1 twoBy adopting the synthetic route of step 1 and using 20d instead of starting material 1b, compound 20e was prepared. LC-MS [M+H] + :334.

[0171] Example 21: Preparation of Compound 21e [ka]

[0172] Using the synthetic route of Example 14, compound 21e was prepared by using iodocyclohexane instead of methyl iodide as a raw material in the third step of the synthesis. LC-MS [M+H] + :376.

[0173] Example 22: Preparation of Compound 22e [ka]

[0174] Using the synthetic route of Example 14, compound 22e was prepared by using 4-bromotetrahydropyran instead of methyl iodide as a raw material in the third step of the synthesis. LC-MS [M+H] + :378.

[0175] Example 23: Preparation of Compound 23e [ka]

[0176] Using the synthetic route of Example 14, compound 23e was prepared by using 4-bromotetrahydrothiopyran instead of methyl iodide as a raw material in the third step of the synthesis. LC-MS [M+H] + :394.

[0177] Example 24: Preparation of Compound 24e [ka]

[0178] Using the synthetic route of Example 14, compound 24e was prepared by using 4-bromotetrahydro-2H-thiopyran 1,1-dioxide instead of methyl iodide as a raw material in the third step of the synthesis. LC-MS: [M+H] + :426.

[0179] Example 25: Preparation of Compound 25e [ka]

[0180] Using the synthetic route of Example 14, compound 25e was prepared by using tert-butyl 4-iodopiperidine-1-carboxylate instead of methyl iodide as a raw material in the third step of the synthesis. LC-MS [M+H] + :477.

[0181] Example 26: Preparation of Compound 26e [ka]

[0182] Using the synthetic route of Example 14, compound 26e was prepared by using iodocyclopentane instead of methyl iodide as a raw material in the third step of the synthesis. LC-MS [M+H] + :362.

[0183] Example 27: Preparation of Compound 27e [ka]

[0184] Using the synthetic route of Example 14, compound 27e was prepared by using iodocyclobutane instead of methyl iodide as a raw material in the third step of the synthesis. LC-MS [M+H] + :348.

[0185] Example 28: Preparation of compound ZB-H-33 [ka]

[0186] The third step in Example 1 About The synthetic route was adopted and compound 20e was used instead of starting material 1c in the third step to prepare the title compound ZB-H-33. 1 H NMR(400MHz,DMSO-d6) δ 13.35(s,1H),7.89(s,2H),7.48(d,J=3.2Hz,1H),7.29(d,J=9.5Hz,1H),6.61(d,J=3.2Hz,1 H),6.08(d,J=9.5Hz,1H),3.05(tt,J=7.2,4.1Hz,1H),1.21-1.11(m,2H),0.91-0.82(m,2H); LC-MS[M+H] + :456.

[0187] Example 29: Preparation of compound ZB-H-34 [ka]

[0188] Adopting the synthetic route of Example 19 and using compound 20e instead of compound 15e, the title compound ZB-H-34 was prepared. 1 H NMR(400MHz,DMSO-d6) δ 13.57(s,1H),11.51(s,1H),8.12(s,2H),7.39(d,J=3.1Hz,1H),7.19(d,J=9.5Hz,1H),6.56(d,J=3.1Hz, 1H),6.07(d,J=9.4Hz,1H),3.03(tt,J=7.1,4.0Hz,1H),1.21-1.12(m,2H),0.91-0.83(m,2H);LC-MS[M+H] + :447.

[0189] Example 30: Preparation of compound ZB-H-38 [ka]

[0190] The third step in Example 1 About The synthetic route was adopted and compound 21e was used instead of starting material 1c in the third step to prepare the title compound ZB-H-38. 1 H NMR(400MHz,DMSO-d6) δ 13.34(s,1H),7.89(s,2H),7.49(d,J=3.2Hz,1H),7.25(d,J=9.4Hz,1H),6.80(d,J=3 .0Hz,1H),6.13(d,J=9.4Hz,1H),2.41-2.28(m,1H),1.95-1.09(m,10H);LC-MS[M+H] + :498.

[0191] Example 31: Preparation of compound ZB-H-44 [ka]

[0192] The third step in Example 1 About Using the same synthetic route, compound 26e was used instead of starting material 1c in the third step to prepare the title compound ZB-H-44. LC-MS [M+H] + :484.

[0193] Example 32: Preparation of compound ZB-H-42 [ka]

[0194] The third step in Example 1 About Using the same synthetic route, compound 27e was used instead of starting material 1c in the third step to prepare the title compound ZB-H-42. LC-MS [M+H] + :470.

[0195] Example 33: Preparation of compound ZB-H-46 [ka]

[0196] The third step in Example 1 ConditionThe synthesis route was adopted, and compound 22e was used instead of starting material 1c in the third step to prepare the title compound ZB-H-46. LC-MS [M+H] + :500.

[0197] Example 34: Preparation of compound ZB-H-48 [ka]

[0198] The third step in Example 1 About The synthetic route was adopted, and compound 23e was used instead of starting material 1c in the third step to prepare the title compound ZB-H-48. LC-MS [M+H] + :516.

[0199] Example 35: Preparation of compound ZB-H-55 [ka]

[0200] The third step in Example 1 About Using the same synthetic route, compound 24e was used instead of starting material 1c in the third step to prepare the title compound ZB-H-49. LC-MS [M+H] + :548.

[0201] Example 36: Preparation of compound ZB-H-50 [ka]

[0202] Using the synthetic route of the third and fourth steps in Example 1, compound 25e was used instead of starting material 1c in the third step, and the Boc protecting group was then removed to prepare the title compound ZB-H-50. LC-MS [M+H] + :499.

[0203] Example 37: Preparation of compound ZB-H-52 [ka]

[0204] First step: Preparation of intermediate 37a

[0205] To a solution of compound 15e (1 mmol) in acetonitrile (4 mL) was added 2-bromoacetonitrile (3.66 mmol, 243 μL), NaI (219 mg, 1.46 mmol), and KCO (202 mg, 1.46 mmol). The mixture was then sealed in a tube and stirred at 100 °C for 16 h. LCMS showed that the starting material was completely consumed. The suspension was filtered through a pad of diatomaceous earth, and the filter cake was washed with ethyl acetate. The combined washings were concentrated to dryness, and compound 37a was obtained by column chromatography isolation and purification. LC-MS [M+H] + :374.

[0206] Second step: Preparation of intermediate 37b

[0207] To a solution of substrate 37a (571.9 mmol) in tetrahydrofuran (3 mL) was added di-tert-butyl dicarbonate (374 mg, 1.72 mmol) and DMAP (70 mg, 571.85 mmol), and the mixture was stirred at 40 °C for 3 h. LCMS showed complete consumption of the starting material, and the required MS was detected. The mixture was partitioned between ethyl acetate and water, and the aqueous layer was extracted with ethyl acetate. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue, which was purified by column chromatography to give compound 37b. LC-MS [M+H] + :474.

[0208] Third Step: Preparation of Intermediate 37c

[0209] To a solution of compound 37b (534.9 mmol) in DMF (3 mL) was added hydroxylamine hydrochloride (297 mg, 4.28 mmol) and NaOAc (351 mg, 4.28 mmol). The mixture was stirred at 80 °C for 1 h. LCMS showed that the starting material was completely consumed. The reaction mixture was concentrated under reduced pressure to remove DMF. The residue was partitioned between ethyl acetate and water, and the aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous NaSO, filtered, and concentrated in vacuo to give compound 37c, which was used directly in the next step without further purification. LC-MS [M+H] + :507.

[0210] Fourth Step: Preparation of Intermediate 37d

[0211] A solution of 37c (520 mmol) in tetrahydrofuran (3 mL) was added with DSC (173 mg) and TEA (105 mg, 1.04 mmol). The mixture was stirred at 60 °C for 16 h. LCMS showed complete consumption of the starting material, and the required MS was detected. The mixture was partitioned between ethyl acetate and water, and the aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue, which was purified by column chromatography to give compound 37d. LC-MS [M+H] + :533.

[0212] Fifth step: Preparation of compound ZB-H-52

[0213] Compound 37d (323.0 mmol) was placed in hydrogen chloride / ethyl acetate (2 mL) and stirred at 25° C. for 2 hours. LCMS showed that the starting material was completely consumed, and the required MS was detected. The mixture was diluted with water (0.5 mL), and the pH was adjusted to 8 with aqueous NaHCO3. The mixture was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue. The crude product was purified by preparative HPLC column to give the title compound ZB-H-52. LC-MS [M+H]+ :433.

[0214] Example 38: Preparation of compound ZB-H-53 [ka]

[0215] First step: Preparation of intermediate 38a

[0216] Compound 15e (1 mmol) was dissolved in acetonitrile (5 mmol), copper(I) iodide (2 mmol) was added, and tert-butyl nitrite (2 mmol) was added dropwise in an ice-water bath. The mixture was stirred overnight at room temperature, and the solvent was evaporated in an evaporator. Compound 38a was obtained by column chromatography isolation and purification. LC-MS: [M+H] + :398.

[0217] Second Step: Preparation of Intermediate 38b

[0218] Compound 38a (1 mmol) was dissolved in dry 1,4-dioxane, and bis(pinacolato)diboron (2 mmol), potassium acetate (3 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (0.05 mmol) were added under argon protection. The mixture was allowed to react at 80 °C overnight. The solvent was evaporated, and compound 38b was obtained by column chromatography. LC-MS: [M+H] + :447.

[0219] Third step: Preparation of compound ZB-H-53

[0220] Compound 38b (1 mmol) and 5-bromo-6-azauracil (38c, 1 mmol) were dissolved together in 1,4-dioxane (5 mL), 2 M aqueous sodium carbonate (2.5 mL) was added, and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (0.1 mmol) was added under argon. The mixture was refluxed at 110 °C for 3 hours under argon, cooled to room temperature, added with 50 mL of water, extracted three times with ethyl acetate, and the organic phases were combined. The organic phase was washed with saturated brine, and the solvent was concentrated under reduced pressure to dryness. Compound ZB-H-53 was isolated and purified by column chromatography. LC-MS [M+H] + :432; 1 H NMR(400MHz,DMSO-d6) δ 12.82(s,1H),12.32(s,1H),8.15(s,2H),7.46(d,J=3.2Hz,1H),7.18(d,J=9.4Hz,1H) ,6.73(d,J=3.2Hz,1H),6.11(d,J=9.4Hz,1H),5.51-5.00(m,1H),1.51(d,J=7.0Hz,6H)

[0221] Example 39: Preparation of compound ZB-H-54 [ka]

[0222] Using the synthetic route of the third step in Example 38, compound 39a was used instead of starting material 38c in the third step to prepare compound ZB-H-54. LC-MS [M+H] + :498. 1 H NMR(400MHz,DMSO-d6) δ 12.52(s,1H),8.20(s,2H),7.46(d,J=3.2Hz,1H),7.19(d,J=9.4Hz,1H),6.74(d,J=3. 1Hz,1H),6.11(d,J=9.4Hz,1H),5.47-5.00(m,1H),3.60(s,3H),1.51(d,J=7.0Hz,6H).

[0223] Example 40: Preparation of compound ZB-H-39 [ka]

[0224] Using the synthetic route of Example 19 and using compound 21e instead of compound 15e, the title compound ZB-H-39 was prepared. LC-MS [M+H] + :488.

[0225] Example 41: Preparation of compound ZB-H-43 [ka]

[0226] Using the synthetic route of Example 19 and using compound 27e instead of starting compound 15e, the title compound ZB-H-43 was prepared. LC-MS [M+H] + :460.

[0227] Example 42: Preparation of compound ZB-H-45 [ka]

[0228] Using the synthetic route of Example 19 and using compound 26e instead of compound 15e, the title compound ZB-H-45 was prepared. LC-MS [M+H] + :474. 1 H NMR(400MHz,DMSO-d6) δ 13.35(s,1H),7.89(s,2H),7.50(d,J=3.2Hz,1H),7.27(d,J=9.4Hz,1H),6.62(d,J=3.2Hz,1H),6 .12(d,J=9.4Hz,1H),5.45-5.04(m,1H),2.27-2.11(m,2H),2.04-1.82(m,4H),1.75-1.60(m,2H).

[0229] Example 43: Preparation of compound ZB-H-47 [ka]

[0230] Using the synthetic route of Example 19 and using compound 22e instead of compound 15e, the title compound ZB-H-47 was prepared. LC-MS [M+H] + :490. 1 H NMR(400MHz,DMSO-d6) δ 13.34(s,1H),7.89(s,2H),7.50(d,J=3.2Hz,1H),7.28(d,J=9.4Hz,1H),6.76(d,J=3.2Hz,1H),6.14(d,J= 9.4Hz,1H),5.22-4.90(m,1H),4.11-3.88(m,2H),3.56-3.51(m,2H),2.76-2.57(m,2H),1.65-1.54(m,2H).

[0231] Example 44: Preparation of compound ZB-H-49 [ka]

[0232] Using the synthetic route of Example 19 and using compound 23e instead of compound 15e, the title compound ZB-H-49 was prepared. LC-MS [M+H] + :506.

[0233] Example 45: Preparation of compound ZB-H-51 [ka]

[0234] Using the synthetic route of Example 19, compound 25e was used instead of compound 15e, and the Boc protecting group was removed to prepare compound ZB-H-51. LC-MS [M+H] + :489.

[0235] Example 46: Preparation of compound ZB-H-56 [ka]

[0236] Using the synthetic route of Example 19 and using compound 24e instead of compound 15e, compound ZB-H-56 was prepared. LC-MS [M+H] + :538.

[0237] Example 47: Preparation of Compound 47b [ka]

[0238] The title compound 47b was prepared by employing the synthetic route of Example 20, except that benzeneboronic acid was used instead of cyclopropylboronic acid in the first step of the synthesis, and the other steps and conditions were the same as those of Example 20. LC-MS: [M+H] + :371.

[0239] Example 48: Preparation of compound ZB-H-57 [ka]

[0240] The third step in Example 1 About Using the same synthetic route, compound 47b was used instead of starting material 1c in the third step to prepare the title compound ZB-H-57. LC-MS: [M+H] + :492.

[0241] Example 49: Preparation of compound ZB-H-58 [ka]

[0242] Using the synthetic route of Example 19 and using compound 47b instead of compound 15e, the title compound ZB-H-58 was prepared. LC-MS [M+H] + :483.

[0243] Example 50: Preparation of Compound 50e [ka]

[0244] The title compound 50e was prepared by the same procedure and conditions as in Example 14, except that the synthetic route of Example 14 was used, except that compound 50a was used instead of the starting material 11a in the first step, and isopropyl iodide was used instead of methyl iodide in the starting material of the third step. LC-MS [M+H] + :338.

[0245] Example 51: Preparation of compound ZB-H-35 [ka]

[0246] The third step in Example 1 About The synthetic route was adopted and compound 50e was used instead of starting compound 1c in the third step to prepare the title compound ZB-H-35. 1 H NMR(400MHz,DMSO-d6) δ LC-MS[M+H] + :459.

[0247] Example 52: Preparation of compound ZB-H-36 [ka]

[0248] Using the synthetic route of Example 19 and using compound 50e instead of compound 15e, the title compound ZB-H-36 was prepared. LC-MS [M+H] + :450.

[0249] Example 53: Preparation of compound ZB-H-59 [ka]

[0250] Using the synthetic route of Example 38, compound 53a was used instead of starting material 38c in the third step to prepare compound ZB-H-59. LC-MS [M+H] + :475.

[0251] Example 54: Preparation of compound ZB-H-76 [ka]

[0252] Step 1: Preparation of Compound 54a

[0253] Compound 50c (500 mg, 1.54 mmol) was dissolved in 1,4-dioxane (10 mL) and placed in a microwave reactor. Benzyl bromide (800 mg, 4.63 mmol) and potassium carbonate (640 mg, 4.63 mmol) were added, and the mixture was reacted at 150°C in a microwave for 12 hours. After the reaction was completed, the solvent was concentrated under reduced pressure to dryness, and compound 54a (1.2 g) was obtained by column chromatography purification. 1 H NMR(600MHz,Chloroform-d) δ 8.38(s,2H),7.78(d,J=1.0Hz,1H),7.42-7.38(m,2H),7.38-7.34(m,2H),7.33 -7.29(m,1H),7.18(dd,J=9.6,0.9Hz,1H),6.73(d,J=9.6Hz,1H),5.37(s,2H). LC-MS:[M+H] + :416.

[0254] Second step: Preparation of compound 54b

[0255] Tin(I) chloride dihydrate (2.7 g, 12.0 mmol) was added to a solution of compound 54a (1 g, 2.41 mmol) in ethanol (20 mL). After the reaction was complete, the mixture was heated to 80 °C and stirred for 6 hours. After the reaction was stopped, the mixture was allowed to cool to room temperature. The solvent was evaporated to dryness under reduced pressure, and then ethyl acetate (200 mL) was added and dissolved. The organic phase was washed three times with aqueous sodium hydroxide (2 M). The organic phase was concentrated and then purified by column chromatography to obtain compound 54b, which was used directly in the next step. 1 H NMR(400MHz,DMSO-d6) δ 8.08(s,1H),7.49(d,J=9.6Hz,1H),7.46-7.41(m,2H),7.39-7.30(m,3H), 6.76(s,2H),6.54(d,J=9.6Hz,1H),6.21(s,2H),5.30(s,2H).LC-MS[M+H] + :386.

[0256] Third step: Preparation of compound 54c

[0257] tert-Butyl nitrite (53 mg, 0.514 mmol) was dissolved in acetic acid (1 mL). At 0 °C, this solution was slowly added to a solution of compound 54b (180 mg, 0.47 mmol) in acetic acid (10 mL) and acetonitrile (10 mL). The mixture was stirred for 30 minutes while maintaining the temperature at 0 °C. A solution of N-cyanoacetylurethane (80 mg, 0.514 mmol) in acetonitrile was added dropwise. After the addition was complete, the mixture was stirred for 3 hours. After the reaction was complete, the reaction mixture was added to saturated aqueous sodium bicarbonate (70 mL). The resulting red solid was filtered, washed with water and petroleum ether, and dried. The product was used directly in the next step. 1H NMR(400MHz,DMSO-d6) δ 12.26(s,1H),11.02(s,1H),8.20(s,1H),8.12(s,2H),7.59(d,J=9.6Hz,1H),7.43(d,J=7.2Hz,2H),7.36(t,J=7.5Hz ,2H),7.33-7.24(m,1H),6.59(d,J=9.6Hz,1H),5.32(s,2H),4.21(q,J=7.1Hz,2H),1.27(t,J=7.1Hz,3H).LC-MS[M+H] + :553.

[0258] Fourth step: Preparation of compound ZB-H-76

[0259] N,N-dimethylacetamide (5 mL) and potassium acetate (2.0 mmol) were added to the solid compound 54c (1.0 mmol), heated to 120 °C, and stirred for 6 hours. After the reaction was completed, the solid was directly subjected to further purification to obtain compound ZB-H-76. 1 H NMR(400MHz,Methanol-d4) δ 8.07(d,J=0.9Hz,1H),7.97(s,2H),7.61(dd,J=9.6,0.9Hz,1H),7.47-7.32(m,5H),6.74(d,J=9.5Hz,1H),5.47(s,2H). LC-MS[M+H] + :507.

[0260] Example 55: Preparation of compound ZB-H-60 [ka]

[0261] Compound ZB-H-60 was prepared by following the synthesis route of Example 54, except that methyl iodide (MeI) was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :431.

[0262] Example 56: Preparation of compound ZB-H-61 [ka]

[0263] Compound ZB-H-61 was prepared by following the synthesis route of Example 54, except that ethyl iodide was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :445.

[0264] Example 57: Preparation of compound ZB-H-62 [ka]

[0265] Compound ZB-H-62 was prepared by following the synthesis route of Example 54, except that iodocyclobutane was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :472.

[0266] Example 58: Preparation of compound ZB-H-63 [ka]

[0267] Compound ZB-H-63 was prepared by following the synthesis route of Example 54, except that iodocyclopentane was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :485.

[0268] Example 59: Preparation of compound ZB-H-64 [ka]

[0269] Following the synthesis route of Example 54, 1-Boc-3-iodopyrrolidine was used instead of benzyl bromide in the first step of Example 54, and the Boc protecting group was then removed to prepare compound ZB-H-64. LC-MS [M+H] + :485.

[0270] Example 60: Preparation of compound ZB-H-65 [ka]

[0271] Compound ZB-H-65 was prepared by following the synthesis route of Example 54, except that iodocyclohexane was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :499.

[0272] Example 61: Preparation of compound ZB-H-66 [ka]

[0273] Compound ZB-H-66 was prepared by following the synthesis route of Example 54, except that 4-iodotetrahydropyran was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :501.

[0274] Example 62: Preparation of compound ZB-H-67 [ka]

[0275] Compound ZB-H-67 was prepared by following the synthesis route of Example 54, except that tetrahydro-4-iodo-2H-thiolane was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :517.

[0276] Example 63: Preparation of compound ZB-H-68 [ka]

[0277] Compound ZB-H-67 (1.0 mmol) was dissolved in dichloromethane (5 mL), and a solution of metachloroperoxybenzoic acid (0.9 mmol) in dichloromethane (5 mL) was added dropwise to the above solution in an ice bath. The reaction was allowed to proceed for 30 minutes in an ice bath. After the reaction was complete, the mixture was quenched with saturated sodium bicarbonate solution, extracted three times with dichloromethane, and the organic phases were combined. The solvent was evaporated in an evaporator, and then purified by column chromatography to give compound ZB-H-68. LC-MS [M+H] + :533.

[0278] Example 64: Preparation of compound ZB-H-69 [ka]

[0279] Compound ZB-H-67 (1.0 mmol) was dissolved in dichloromethane (5 mL), and a solution of metachloroperoxybenzoic acid (2.0 mmol) in dichloromethane (5 mL) was added dropwise to the above solution in an ice bath. The reaction was allowed to proceed for 30 minutes in an ice bath. After the reaction was complete, the mixture was quenched with saturated sodium bicarbonate solution, extracted three times with dichloromethane, and the organic phases were combined. The solvent was evaporated using an evaporator, and then purified by column chromatography to give compound ZB-H-69. LC-MS [M+H] + :549.

[0280] Example 65: Preparation of compound ZB-H-70 [ka]

[0281] Compound ZB-H-70 was prepared by following the synthetic routes of Examples 47 and 54. LC-MS [M+H] + :593.

[0282] Example 66: Preparation of compound ZB-H-71 [ka]

[0283] Step 1: Preparation of Compound 66a

[0284] Compound 50c (200 mg, 0.615 mmol) was dissolved in 1,4-dioxane (10 mL) and placed in a microwave reactor. Bromomethylcyclopropane (250 mg, 1.845 mmol) and potassium carbonate (170 mg, 1.231 mmol) were added, respectively, and the mixture was reacted at 150 °C in a microwave for 12 hours. After the reaction was completed, the solvent was concentrated under reduced pressure to dryness, and compound 66a was obtained by column chromatography purification. 1 H NMR(400MHz,Chloroform-d) δ 8.37(s,2H),7.88(s,1H),7.16(d,J=9.6Hz,1H),6.67(d,J=9.6Hz,1H),4.04(d,J=7.1Hz,2H),2.02(m,1H),1.30-1.15(m,4H). LC-MS:[M+H] + :379.

[0285] Second step: Preparation of compound 66b

[0286] To a solution of compound 66a (80 mg, 0.212 mmol) in ethanol (10 mL) was added tin(I) chloride dihydrate (238 mg, 1.058 mmol). After the addition was complete, the mixture was heated to 80 °C and stirred for 6 h. After the reaction was stopped, the mixture was allowed to cool to room temperature. The solvent was evaporated to dryness under reduced pressure, and then ethyl acetate (10 mL) was added and dissolved. The organic phase was washed three times with aqueous sodium hydroxide (2 M, 10 mL). The organic phase was concentrated and then purified by column chromatography to give compound 66b, which was used directly in the next step. 1 H NMR(400MHz,Methanol-d4) δ 8.01(s,1H),7.39(d,J=9.5Hz,1H),6.74(s,2H),6.58(d,J=9.5Hz,1H),4.07(d,J=7.1Hz,2H),1.95(m,1H),0.57-0.46(m,4H). LC-MS[M+H] + :350.

[0287] Third step: Preparation of compound 66c

[0288] tert-Butyl nitrite (16.2 mg, 0.1573 mmol) was dissolved in acetic acid (2 mL). This solution was slowly added to a solution of compound 66b (50 mg, 0.143 mmol) in acetic acid (2 mL) and acetonitrile (2 mL) at 0 °C and stirred for 30 min at 0 °C. A solution of N-cyanoacetylurethane (25 mg, 0.1573 mmol) in acetonitrile (2 mL) was added dropwise. After the addition was complete, the mixture was stirred for 3 h. After the reaction was complete, the reaction mixture was added to saturated aqueous sodium bicarbonate (20 mL) and filtered to give a red solid. The solid was washed with water (10 mL) and petroleum ether (10 mL), dried, and used directly in the next step. 1 H NMR(400MHz,DMSO-d6) δ 12.28(s,1H),11.02(s,1H),8.25(d,J=0.9Hz,1H),8.13(s,2H),7.54(dd,J=9.6,0.9Hz,1H),6.51(d,J=9.6 Hz,1H),4.21(q,J=7.1Hz,2H),3.99(d,J=7.1Hz,2H),1.27(t,J=7.1Hz,3H),1.23(m,1H),0.52-0.44(m,4H). LC-MS[M+H] + :517.

[0289] Fourth step: Preparation of compound ZB-H-71

[0290] Compound 66c (81 mg, 0.1571 mmol) was added to N,N-dimethylacetamide (5 mL) and potassium acetate (17 mg, 0.1716 mmol), heated to 120 °C, and stirred for 6 hours. After the reaction was completed, the mixture was directly subjected to further purification to obtain compound ZB-H-71. 1 H NMR(400MHz,Methanol-d4) δ 8.19(d,J=0.9Hz,1H),7.96(s,2H),7.56(dd,J=9.6,0.9Hz,1H),6.66(d,J =9.5Hz,1H),4.14(d,J=7.1Hz,2H),1.48-1.36(m,1H),0.64-0.49(m,4H). LC-MS[M+H]+ :471.

[0291] Example 67: Preparation of compound ZB-H-72 [ka]

[0292] Compound ZB-H-72 was prepared by following the synthesis route of Example 54, except that (iodomethyl)cyclobutane was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :485.

[0293] Example 68: Preparation of compound ZB-H-73 [ka]

[0294] Compound ZB-H-73 was prepared by following the synthesis route of Example 54, except that (iodomethyl)cyclopentane was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :489.

[0295] Example 69: Preparation of compound ZB-H-74 [ka]

[0296] Step 1: Synthesis of compound 69a

[0297] Compound 69a was obtained by following the first step of the synthetic route in Example 54, except that compound 14c was used instead of compound 50c. 1 H NMR(400MHz,Chloroform-d) δ 8.38(s,2H),7.40-7.27(m,5H),7.03(dd,J=9.6,0.8Hz,1H),6.92(d,J=3.3Hz,1 H),6.49(d,J=9.5Hz,1H),6.37(dd,J=3.3,0.8Hz,1H),5.41(s,2H).LC-MS[M+H]+ :415.

[0298] Second step: Synthesis of compound 69b

[0299] Compound 69b was obtained by following the second step of the synthetic route in Example 54. 1 H NMR(400MHz,DMSO-d6) δ 7.42-7.37(m,2H),7.33(t,J=7.4Hz,2H),7.28-7.22(m,2H),7.18(d,J=9.4Hz,1H),6.75( s,2H),6.46(d,J=3.2Hz,1H),6.18(d,J=9.4Hz,1H),6.08(s,2H),5.28(s,2H).LC-MS[M+H] + :385.

[0300] Third step: Synthesis of compound 69c

[0301] Compound 69c was obtained by following the third step of the synthetic route in Example 54. LC-MS [M+H] + :552.

[0302] Fourth step: Synthesis of compound ZB-H-74

[0303] Compound ZB-H-74 was obtained by following the fourth step of the synthetic route in Example 54. 1 H NMR(400MHz,Methanol-d4) δ 7.92(s,2H),7.40-7.28(m,7H),6.56(dd,J=3.3,0.8Hz,1H),6.45(d,J=9.4Hz,1H),5.48(s,2H)..LC-MS[M+H] + :506.

[0304] Example 70: Preparation of compound ZB-H-75 [ka]

[0305] Step 1: Synthesis of compound 70a

[0306] Compound 50c (500 mg, 1.54 mmol) was dissolved in toluene (20 mL), and potassium cyclopropyltrifluoroborate (4.62 mmol), copper acetate (0.385 mmol), potassium carbonate (3.08 mmol), 1,10-phenanthroline (0.1925 mmol), and water (2 mL) were added. The mixture was purged with oxygen gas three times and reacted at 80°C overnight. After completion of the reaction, the solvent was concentrated under reduced pressure to dryness, and compound 70a was obtained by column chromatography purification. 1 H NMR(400MHz,Chloroform-d) δ 8.42(s,2H),8.07(d,J=0.9Hz,1H),7.14(dd,J=9.7,0.9Hz,1H),6.62(d,J=9.6Hz, 1H),3.20(tt,J=7.3,4.0Hz,1H),1.37-1.30(m,2H),1.14-1.08(m,2H).LC-MS[M+H] + :366.

[0307] Second step: synthesis of compound 70b

[0308] Compound 70b was obtained by following the second step of the synthetic route in Example 54. 1 H NMR(600MHz,DMSO-d6) δ 8.00(d,J=0.9Hz,1H),7.38(dd,J=9.5,0.9Hz,1H),6.76(s,2H),6.38(d,J=9.5Hz,1H),6.27-6.03( br.s,2H),3.10(tt,J=7.3,4.0Hz,1H),1.16(td,J=7.4,5.6Hz,2H),0.93-0.87(m,2H).LC-MS[M+H] + :336.

[0309] Third step: Synthesis of compound 70c

[0310] Compound 70c was obtained by following the third step of the synthetic route in Example 54. 1H NMR(400MHz,DMSO-d6) δ 12.27(s,1H),11.00(s,1H),8.12(s,J=1.9Hz,2H),8.10(m,1H),7.49(d,J=9.6Hz,1H),6.43(d,J=9.6Hz,1H),4.21(q ,J=7.1Hz,2H),3.12(tt,J=7.2,3.9Hz,1H),1.27(t,J=7.1Hz,3H),1.23-1.13(m,2H),0.94-0.86(m,2H).LC-MS[M+H] + :503.

[0311] Fourth step: Synthesis of compound ZB-H-75

[0312] Compound ZB-H-75 was obtained by following the fourth step of the synthetic route in Example 54. 1 H NMR(500MHz,DMSO-d6) δ 8.18(d,J=0.9Hz,1H),7.93(s,2H),7.63(d,J=9.6Hz,1H),6.44(d,J=9.6Hz,1H),3.1 3(td,J=7.1,3.7Hz,1H),1.18(td,J=7.4,5.6Hz,2H),0.95-0.88(m,2H).LC-MS[M+H] + :457.

[0313] Example 71: Preparation of compound ZB-H-77 [ka]

[0314] Step 1: Preparation of Compound 71a

[0315] Compound 50c (200 mg, 0.615 mmol) was dissolved in 1,4-dioxane (10 mL) and placed in a microwave reactor. 4-fluorobenzyl bromide (233 mg, 1.231 mmol) and potassium carbonate (170 mg, 1.231 mmol) were added, and the mixture was reacted at 150°C in a microwave for 12 hours. After the reaction was completed, the solvent was concentrated under reduced pressure to dryness, and compound 71a was obtained by column chromatography purification. 1H NMR(400MHz,Methanol-d4) δ 6.98(s,2H),6.57(d,J=0.9Hz,1H),6.04(dd,J=9.5,0.9Hz,1H),5.92-5.86(m,2H),5.57-5.49(m,2H),5.14(d,J=9.5Hz,1H),3.85(s,2H). LC-MS:[M+H] + :434.

[0316] Second step: Preparation of compound 77b

[0317] Tin(I) chloride dihydrate (238 mg, 1.058 mmol) was added to a solution of compound 77a (92 mg, 0.212 mmol) in ethanol (10 mL). After the reaction was complete, the mixture was heated to 80 °C and stirred for 6 hours. After the reaction was stopped, the mixture was allowed to cool to room temperature. The solvent was evaporated to dryness under reduced pressure, and then ethyl acetate (10 mL) was added and dissolved. The organic phase was washed three times with aqueous sodium hydroxide (2 M, 10 mL). The organic phase was concentrated and then purified by column chromatography to obtain compound 71b, which was used directly in the next step. LC-MS [M+H] + :404.

[0318] Third step: Preparation of compound 71c

[0319] tert-Butyl nitrite (16.2 mg, 0.1573 mmol) was dissolved in acetic acid (2 mL). At 0 °C, this solution was slowly added to a solution of compound 71b (58 mg, 0.143 mmol) in acetic acid (2 mL) and acetonitrile (2 mL). The mixture was stirred for 30 minutes at 0 °C. A solution of N-cyanoacetylurethane (25 mg, 0.1573 mmol) in acetonitrile (2 mL) was added dropwise. After the addition was complete, the mixture was stirred for 3 hours. After the reaction was complete, the reaction mixture was added to saturated aqueous sodium bicarbonate (20 mL). The resulting red solid was filtered, washed with water (10 mL) and petroleum ether (10 mL), and dried. The product was used directly in the next step. 1H NMR(400MHz,DMSO-d6) δ 12.27(s,1H),11.01(s,1H),8.25(d,J=0.9Hz,1H),8.12(s,2H),7.59(d,J=9.6Hz,1H),7.56-7.47(m,2 H),7.23-7.16(m,2H),6.58(d,J=9.6Hz,1H),5.30(s,2H),4.21(q,J=7.1Hz,2H),1.27(t,J=7.1Hz,3H). LC-MS[M+H] + :571.

[0320] Fourth step: Preparation of compound ZB-H-77

[0321] Compound 71c (89 mg, 0.157 mmol) was added to N,N-dimethylacetamide (5 mL) and potassium acetate (17 mg, 0.1716 mmol), heated to 120 °C, and stirred for 6 hours. After the reaction was completed, the solid was directly subjected to further purification to obtain compound ZB-H-77. 1 H NMR(400MHz,DMSO-d6) δ 13.36(s,1H),8.32(s,1H),7.93(s,2H),7.74(d,J=9.6Hz,1H),7.56-7.47(m,2H),7.20(m,2H),6.59(d,J=9.6Hz,1H),5.31(s,2H).LC-MS[M+H] + :525.

[0322] Example 72: Preparation of compound ZB-H-78 [ka]

[0323] Compound ZB-H-78 was prepared by following the synthesis route of Example 54, except that 3-fluorobenzyl bromide was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :525.

[0324] Example 73: Preparation of compound ZB-H-79 [ka]

[0325] Compound ZB-H-79 was prepared by following the synthesis route of Example 54, except that 4-methylbenzyl bromide was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :521.

[0326] Example 74: Preparation of compound ZB-H-80 [ka]

[0327] Compound ZB-H-80 was prepared by following the synthesis route of Example 54, except that 4-methoxybenzyl bromide was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :537.

[0328] Example 75: Preparation of compound ZB-H-81 [ka]

[0329] Step 1: Synthesis of compound 75a

[0330] Compound 75a was obtained by following the first step of the synthetic route in Example 54, except that (bromomethyl)cyclohexane was used instead of benzyl bromide. 1 H NMR(600MHz,Chloroform-d) δ 8.36(s,2H),7.82(s,1H),7.13(d,J=9.6Hz,1H),6.62(d,J=9.6Hz,1H),3.95(d,J=7.4Hz, 2H),1.97(dqt,J=10.6,6.9,3.2Hz,1H),1.78-1.60(m,5H),1.24-1.11(m,5H).LC-MS[M+H] + :422.

[0331] Second step: Synthesis of compound 75b

[0332] Compound 75b was obtained by following the second step of the synthetic route in Example 54. 1 H NMR(400MHz,Methanol-d4) δ 7.88(s,1H),7.36(d,J=9.6Hz,1H),6.78(s,2H),6.63(d,J=9.6Hz,1H),4.03(d,J=7.3Hz,2H),2.00(m,1H),1.73(m,5H),1.24(m,5H). LC-MS[M+H] + :392.

[0333] Third step: Synthesis of compound 75c

[0334] Compound 75c was obtained by following the third step of the synthetic route in Example 54. 1 H NMR(600MHz,DMSO-d6) δ 12.26(s,1H),11.01(s,1H),8.21(s,1H),8.13(s,2H),7.52(d,J=9.6Hz,1H),6.49(d,J=9.6Hz,1H),4.21( q,J=7.1Hz,2H),3.93(d,J=7.3Hz,2H),1.90(m,1H),1.71-1.57(m,5H),1.27(t,J=7.1Hz,3H),1.14(m,5H). LC-MS[M+H] + :559.

[0335] Fourth step: Synthesis of compound ZB-H-81

[0336] Compound ZB-H-81 was obtained by following the fourth step of the synthetic route in Example 54. 1 H NMR(600MHz,DMSO-d6) δ 13.35(s,1H),8.28(s,1H),7.93(s,2H),7.67(d,J=9.5Hz,1H),6.51(d,J=9.5Hz,1H),3.94(d,J= 7.3Hz,2H),1.90(dtt,J=12.9,8.9,4.6Hz,1H),1.71-1.58(m,5H),1.19-1.07(m,5H).LC-MS[M+H] + :513.

[0337] Example 76: Preparation of compound ZB-H-82 [ka]

[0338] Compound ZB-H-82 was prepared by following the synthesis route of Example 54, except that 4-bromomethyltetrahydropyran was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :515.

[0339] Example 77: Preparation of compound ZB-H-83 [ka]

[0340] Compound ZB-H-83 was prepared by following the synthesis route of Example 54, except that 4-(bromomethyl)-tetrahydro-2H-thiolane was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :531.

[0341] Example 78: Preparation of compound ZB-H-84 [ka]

[0342] Compound ZB-H-83 (1.0 mmol) was dissolved in dichloromethane (5 mL), and a solution of metachloroperoxybenzoic acid (2.0 mmol) in dichloromethane (5 mL) was added dropwise to the above solution in an ice bath. The reaction was allowed to proceed for 30 minutes in an ice bath. After the reaction was complete, the mixture was quenched with saturated sodium bicarbonate solution, extracted three times with dichloromethane, and the organic phases were combined. The solvent was evaporated using an evaporator, and then purified by column chromatography to give compound ZB-H-84. LC-MS [M+H] + :563.

[0343] Example 79: Preparation of compound ZB-H-85 [ka]

[0344] Compound ZB-H-85 was prepared by following the synthesis route of Example 54, except that 4-(bromomethyl)-pyridine was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :508.

[0345] Example 80: Preparation of compound ZB-H-86 [ka]

[0346] Compound ZB-H-86 was prepared by following the synthesis route of Example 54, except that 3-bromomethylthiophene was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :513.

[0347] Example 81: Preparation of compound ZB-H-87 [ka]

[0348] Compound ZB-H-87 was prepared by following the synthesis route of Example 54, except that 2-bromomethylthiophene was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :513.

[0349] Example 82: Preparation of compound ZB-H-88 [ka]

[0350] Compound ZB-H-88 was prepared by following the synthesis route of Example 54, except that 2-bromomethylfuran was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :497.

[0351] Example 83: Preparation of compound ZB-H-89 [ka]

[0352] Compound ZB-H-89 was prepared by following the synthesis route of Example 54, except that 3-(bromomethyl)-5-methylisoxazole was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :512.

[0353] Example 84: Preparation of compound ZB-H-90 [ka]

[0354] Following the synthesis route of Example 54, compound ZB-H-90 was prepared by using 4-(bromomethyl)-pyridine instead of benzyl bromide in the first step of Example 54 and compound 14c instead of compound 50c. LC-MS [M+H] + :507.

[0355] Example 85: Preparation of compound ZB-H-91 [ka]

[0356] Following the synthesis route of Example 54, compound ZB-H-91 was prepared by using 3-bromomethylthiophene instead of benzyl bromide in the first step of Example 54 and compound 14c instead of compound 50c. LC-MS [M+H] + :512.

[0357] Example 86: Preparation of compound ZB-H-92 [ka]

[0358] Following the synthesis route of Example 54, compound ZB-H-92 was prepared by using 2-bromomethylthiophene instead of benzyl bromide in the first step of Example 54 and compound 14c instead of compound 50c. LC-MS [M+H] + :512.

[0359] Example 87: Preparation of compound ZB-H-93 [ka]

[0360] Following the synthesis route of Example 54, compound ZB-H-93 was prepared by using 2-bromomethylfuran instead of benzyl bromide in the first step of Example 54 and compound 14c instead of compound 50c. LC-MS [M+H] + :496.

[0361] Example 88: Preparation of compound ZB-H-94 [ka]

[0362] Following the synthesis route of Example 54, compound ZB-H-94 was prepared by using 3-(bromomethyl)-5-methylisoxazole instead of benzyl bromide in the first step of Example 54 and compound 14c instead of compound 50c. LC-MS [M+H] + :511.

[0363] Example 89: Preparation of compound ZB-H-95 [ka]

[0364] Step 1: Synthesis of compound 89a

[0365] Compound 89a was obtained by following the first step of the synthetic route in Example 54, except that 1-iodo-2-methylpropane was used instead of benzyl bromide. 1H NMR(600MHz,Chloroform-d) δ 8.37(s,2H),7.82(s,1H),7.14(d,J=9.6Hz,1H),6.63(d,J=9.6Hz,1H),3. 95(d,J=7.6Hz,2H),1.28-1.15(m,1H),1.00(d,J=6.7Hz,6H).LC-MS[M+H] + :382.

[0366] Second step: Synthesis of compound 89b

[0367] Compound 89b was obtained by following the second step of the synthetic route in Example 54. 1 H NMR(400MHz,Methanol-d4) δ 8.02(s,1H),7.42(d,J=9.5,1H),6.78(s,2H),6.61(d,J=9.5Hz,1H),4.03(d,J=7.6Hz,2H),2.31(m,1H),0.99(d,J=6.7Hz,6H).LC-MS[M+H] + :352.

[0368] Third step: Synthesis of compound 89c

[0369] Compound 89c was obtained by following the third step of the synthetic route in Example 54. 1 H NMR(600MHz,DMSO-d6) δ 8.22(s,1H),8.12(s,2H),7.53(d,J=9.6Hz,1H),6.50(d,J=9.6Hz,1H),4.21(q,J=7.1Hz,2H),3. 92(d,J=7.5Hz,2H),2.28-2.16(m,1H),1.27(t,J=7.1Hz,3H),0.92(d,J=6.7Hz,6H).LC-MS[M+H] + :519.

[0370] Fourth step: Synthesis of compound ZB-H-95

[0371] Compound ZB-H-95 was obtained by following the fourth step of the synthetic route in Example 54. 1H NMR(600MHz,DMSO-d6) δ 13.35(s,1H),8.29(d,J=1.0Hz,1H),7.93(s,2H),7.68(d,J=9.6Hz,1H),6.51(d,J =9.6Hz,1H),3.93(d,J=7.5Hz,2H),2.21(m,1H),0.93(d,J=6.7Hz,6H).LC-MS[M+H] + :473.

[0372] Example 90: Preparation of compound ZB-H-96 [ka]

[0373] Compound ZB-H-96 was prepared by following the synthesis route of Example 54, except that 4-chlorobenzyl bromide was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :541.

[0374] Example 91: Preparation of compound ZB-H-97 [ka]

[0375] Compound ZB-H-97 was prepared by following the synthesis route of Example 54, except that 3-chlorobenzyl bromide was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :541.

[0376] Example 92: Preparation of compound ZB-H-98 [ka]

[0377] Compound ZB-H-98 was prepared by following the synthesis route of Example 54, except that 2-chlorobenzyl bromide was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :541.

[0378] Example 93: Preparation of compound ZB-H-99 [ka]

[0379] Compound ZB-H-99 was prepared by following the synthesis route of Example 54, except that 2-fluorobenzyl bromide was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :525.

[0380] Example 94: Preparation of compound ZB-H-100 [ka]

[0381] By referring to the synthetic route of Example 54, compound ZB-H-100 was prepared by using 2,4-difluorobenzyl bromide instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :543.

[0382] Example 95: Preparation of compound ZB-H-101 [ka]

[0383] Compound ZB-H-101 was prepared by following the synthesis route of Example 54, except that 1-bromo-trifluoro-p-xylene was used instead of benzyl bromide in the first step of Example 54. LC-MS [M+H] + :575.

[0384] Example 96: THR reporter gene assay for agonist activity of compounds on THRα, THRβ

[0385] Huh7 cells were cultured in DMEM medium containing 10% FBS. Cells were seeded into 10 cm cell culture dishes and allowed to grow to approximately 90% fullness. Then, using liposomal Lipofectamine 2000, they were co-transfected with a human THRα eukaryotic expression plasmid or a human THRβ eukaryotic expression plasmid and a reporter plasmid containing a THR response element driven by PGL4.26-DR4-Luc. The procedure was described in the Lipofectamine 2000 instructions. The day after transfection, phenol red-free DMEM medium (containing 5% charcoal-stripped FBS) was seeded into 96-well cell culture plates at a density of 20,000 cells per well in a volume of 135 μL per well. Six hours after seeding, cells were allowed to form a wall. Compounds dissolved in DMSO were diluted 20-fold with phenol red-free DMEM medium (containing 5% charcoal-stripped FBS) to a final concentration of 10x and added to cell wells at 15 μL / well; the compounds were again diluted 10-fold to the final concentration. Triiodothyronine T3 (100 nM) was used as a positive control, and 0.5% DMSO was used as a blank control. After administration, cells were incubated overnight (16 hours) at 37°C in a 5% CO2 incubator. After incubation, the medium was discarded, and 35 μL of serum-free phenol red-free DMEM medium was added to each well. 35 μL of Steady-Glo was added, and the samples were shaken at room temperature for 10 minutes in the dark, after which the chemiluminescence values ​​of the samples were measured.

[0386] The agonist activity of the compound was calculated as follows: % Effect = (compound - blank control) / (positive control - blank control) x 100%. EC of the compound 50 was calculated by fitting the logarithm of the agonist activity of the compound to the compound concentration using GraphPad Prism. A lower EC50 value indicates better activity.

[0387] EC of agonist activity of compounds for THRα and THRβ 50The values ​​were first corrected by the EC50 values ​​of the agonist activity of T3 on THRα and THRβ in the same experiment, respectively, and the resulting values ​​were used to calculate the fold selectivity, i.e., receptor selectivity. Specifically, it was calculated as follows: Selectivity = (Compound THRα EC 50 / T3 THRα EC 50 ) / (compound THRβ EC 50 / T3 THRβ EC 50 ) The higher the value, the more selective the compound is for the THRβ receptor.

[0388] Resmetirom (MG1-3196), a highly selective THR-β agonist, was used as a positive control (Reference: J Med Chem. 2014, 57(10):3912-3923). [Table 1] JPEG0007729891000122.jpg213159

[0389] The experimental results showed that the activity or selectivity of some compounds having the structures of the present disclosure (e.g., ZB-H-02, ZB-H-17, ZB-H-18, ZB-H-25, ZB-H-26, ZB-H-33, ZB-H-38, ZB-H-74, ZB-H-75, ZB-H-76, ZB-H-81, ZB-H-95, etc.) was significantly higher than that of the positive compound, and the activity or selectivity of most of the other compounds was comparable to that of the positive compound (MG1-3196).

[0390] Example 97: Effect of reducing serum cholesterol levels measured in ICR mice after a single administration of the compound

[0391] ICR mice were used as test animals, and changes in serum cholesterol levels were measured after intragastric administration of a compound. ICR mice (male, 7-8 weeks old) were randomly divided into groups according to body weight. The vehicle control group received intragastric administration of 0.25% CMC-Na, while the compound group received a single dose of the corresponding compound to be tested. On the day of administration, the mice were fasted overnight. 24 hours after administration, blood was collected and serum was obtained. Cholesterol levels in the serum of the mice were measured using a Total Cholesterol Assay Kit (Zhejiang Dong'ou Diagnostic Products Co., Ltd.). The percentage reduction in serum cholesterol levels in each group of mice was calculated, with the vehicle control group set at 100%. [Table 2] JPEG0007729891000124.jpg75159

[0392] Experimental results 24 hours after administration confirmed that the compound had a certain effect in the animal body.

Claims

1. A compound represented by general formula (Ic): 【Chemical 1】 wherein ring B is 【Chemistry 2】 R １ is hydrogen, substituted or unsubstituted C １－１０ alkyl group, substituted or unsubstituted C ３－１０ Cycloalkyl groups, substituted or unsubstituted 3- to 10-membered heterocycloalkyl groups, substituted or unsubstituted C ６－１０ and a substituted or unsubstituted 5- to 10-membered heteroaryl group, wherein the substituent is a halogen atom, a hydroxy group, ═O, C １－６ Alkoxy group, C １－６ Alkyl group, C ３－１０ Cycloalkyl group, C ６－１０ Aryl group, halo C ６－１０ Aryl group, C １－１０ Alkyl group C ６－１０ Aryl group, C １－１０ Alkoxy group C ６－１０ Aryl group, 5-10 membered heteroaryl group, C １－１０ alkyl group, 5-10 membered heteroaryl group, halo 5-10 membered heteroaryl group, 3-10 membered heterocycloalkyl group and —NR １０ R １１ selected from the group consisting of: R ２ and R ３ each independently represents a halogen atom, a substituted or unsubstituted C １－６ Alkyl groups and substituted or unsubstituted C ３－６ The substituents of the substituted groups are selected from the group consisting of halogen atoms, hydroxy groups, C １－４ Alkyl group and C １－４ alkoxy groups; R ４ is hydrogen, a cyano group, -NR １０ R １１ , substituted or unsubstituted C １－６ alkyl group, substituted or unsubstituted C ３－６ Cycloalkyl groups and substituted or unsubstituted C ２－８ alkynyl groups, and the substituents of the substituted groups are selected from the group consisting of halogen atoms, hydroxy groups, cyano groups, and C １－６ alkoxy groups; R ６ is hydrogen and substituted or unsubstituted C １－６ alkyl groups, and the substituents of the substitution are selected from the group consisting of halogen atoms, hydroxy groups and C １－６ alkoxy groups; L is -NR １０ C(O)—; Each R １０ are independently hydrogen and substituted or unsubstituted C １－３ alkyl groups, and the substituents of the substitution are selected from the group consisting of halogen atoms, hydroxy groups and C １－３ alkoxy groups; each R １１ are independently hydrogen and substituted or unsubstituted C １－６ alkyl groups, and the substituents of the substitution are selected from the group consisting of halogen atoms, hydroxy groups and C １－６ alkoxy groups; X １ and X ２ are each independently N and CR １２ and R １２ is hydrogen, halogen, cyano group, substituted or unsubstituted C １－４ alkyl group, substituted or unsubstituted C ３－６ cycloalkyl groups, wherein the substituents are selected from the group consisting of halogen atoms, hydroxy groups, and C １－６ alkoxy groups; X ３ is CH, X ４ However, N and CR １３ and R １３ is hydrogen, substituted or unsubstituted C １－４ Alkyl groups and substituted or unsubstituted C ３－６ cycloalkyl groups, wherein the substituents are selected from the group consisting of halogen atoms, hydroxy groups, and C １－６ selected from the group consisting of alkoxy groups; A compound, its pharmacologically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, crystalline polymorph, solvate or isotopically labeled compound.

2. The compound of general formula (Ic) is selected from the group consisting of compounds represented by general formula (Id), 【Chemistry 3】 R ２ and R ３ are each independently selected from the group consisting of halogen atoms; L is —NHC(O)—; X １ and X ２ are each independently N and CR １２ and R １２ is hydrogen, halogen, cyano group, substituted or unsubstituted C １－４ alkyl group, substituted or unsubstituted C ３－６ cycloalkyl groups, wherein the substituents are selected from the group consisting of halogen atoms, hydroxy groups, and C １－６ alkoxy groups; R １ is the same as defined in general formula Ic of claim 1.

2. The compound according to claim 1, or a pharmacologically acceptable salt thereof, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, crystalline polymorph, solvate, or isotope-labeled compound.

3. The compound of general formula (Ic) is selected from the group consisting of compounds represented by general formula (Ie), 【Chemistry 4】 R ２ and R ３ are each independently selected from the group consisting of halogen atoms; X １ and X ２ are each independently N and CR １２ and R １２ is hydrogen, halogen, cyano group, substituted or unsubstituted C １－４ alkyl group, substituted or unsubstituted C ３－６ cycloalkyl groups, wherein the substituents are selected from the group consisting of halogen atoms, hydroxy groups, and C １－６ alkoxy groups; R １ and R ４ is the same as defined in general formula Ic of claim 1.

2. The compound according to claim 1, or a pharmacologically acceptable salt thereof, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, crystalline polymorph, solvate, or isotope-labeled compound.

4. The compound of general formula (Ic) is selected from the group consisting of compounds represented by general formula (If), 【Chemistry 5】 R ２ and R ３ are each independently selected from the group consisting of halogen atoms; X １ and X ２ are each independently N and CR １２ and R １２ is hydrogen, halogen, cyano group, substituted or unsubstituted C １－４ alkyl group, substituted or unsubstituted C ３－６ cycloalkyl groups, wherein the substituents are selected from the group consisting of halogen atoms, hydroxy groups, and C １－６ alkoxy groups; R １ and R ６ is the same as defined in general formula Ic of claim 1.

2. The compound according to claim 1, or a pharmacologically acceptable salt thereof, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, crystalline polymorph, solvate, or isotope-labeled compound.

5. A compound, a pharmacologically acceptable salt thereof, a stereoisomer, an enantiomer, a diastereomer, an atropisomer, a racemate, a crystalline polymorph, a solvate, or an isotopically labeled compound thereof, selected from the group consisting of: 【Chemistry 6】 【change】 【change】 【change】 【change】 【change】 【change】

6. A pharmaceutical composition comprising one or more compounds selected from the group consisting of the compound according to any one of claims 1 to 5, its pharmacologically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, crystalline polymorph, solvate, and isotope-labeled compound, and any pharmacologically acceptable additive.

7. Use of the compound according to any one of claims 1 to 5, a pharmacologically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, crystalline polymorph, solvate or isotope-labeled compound thereof, or the pharmaceutical composition according to claim 6, in the manufacture of a medicament for treating a metabolic-related disease.

8. The metabolic disease is selected from the group consisting of obesity, hyperlipidemia, hypercholesterolemia, diabetes, non-alcoholic fatty liver disease, hepatic steatosis, atherosclerosis, hypothyroidism, and thyroid cancer, and particularly, the metabolic disease is selected from the group consisting of non-alcoholic fatty liver disease (NASH), hypothyroidism, and thyroid cancer.

8. Use according to claim 7.

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