Novel thyroid hormone beta receptor agonists

Abstract

The present application provides a novel thyroid hormone β receptor agonist of formula (I) having better activity, selectivity or safety and the use thereof for preventing or treating diseases related to the action of β receptor agonists, including, for example, obesity, hyperlipidemia, hypercholesterolemia, diabetes, liver diseases (fatty liver, NASH, NAFLD, etc.), cardiovascular diseases (atherosclerosis, etc.), thyroid diseases (hypothyroidism, thyroid cancer, etc.), and the like.

Description

Technical Field

[0001] This application relates to a novel thyroid hormone β-receptor agonist, which can be used to treat obesity, hyperlipidemia, hypercholesterolemia, diabetes, liver diseases (fatty liver, NASH, NAFLD, etc.), cardiovascular diseases (atherosclerosis, etc.), and thyroid diseases (hypothyroidism, thyroid cancer, etc.). Background Technology

[0002] Thyroid hormones are secreted by the thyroid gland and act on almost all cells in the human body. These include thyroxine (T4) and triiodothyronine (T3). T4 is deiodinated to T3 by specific deiodinases, thus becoming effective. T3 has a rapid and strong effect, but its duration of action is shorter than that of T4, while T4 has a slower and weaker effect, but its duration of action is longer. Specific deiodinases are present in all tissues, but are more abundant in the liver and kidneys.

[0003] Thyroid hormones are essential for normal human growth and development; both insufficient and excessive secretion can cause disease. Hypothyroidism affects both physical and intellectual development, potentially leading to cretinism. In adults, hypothyroidism can cause myxedema. Hyperthyroidism causes symptoms such as nervousness, irritability, tremors, increased heart rate, and increased cardiac output. Thyroid hormones promote oxidation, increase oxygen consumption, raise the basal metabolic rate, and increase heat production.

[0004] Under normal circumstances, under the regulation of the central nervous system, the hypothalamus releases thyroid-stimulating hormone (TRH) to regulate the secretion of thyroid-stimulating hormone (TSH) from the anterior pituitary gland. TSH, in turn, stimulates thyroid cells to secrete T4 and T3. When the concentrations of T4 and T3 in the blood increase, negative feedback inhibits the synthesis and release of TSH from the anterior pituitary gland, reducing the pituitary's responsiveness to TRH and thus decreasing TSH secretion, preventing excessive thyroid hormone secretion. Conversely, when the concentrations of T4 and T3 in the blood decrease, the negative feedback effect on the anterior pituitary weakens. Increased TSH secretion then promotes increased secretion of T4 and T3. In summary, the hypothalamic-pituitary-thyroid regulatory loop maintains a relatively constant level of thyroid hormone secretion.

[0005] The biological activity of thyroid hormones is mediated by thyroid hormone receptors (TRs). Thyroid hormone receptors belong to the superfamily of nuclear receptors. TRs possess ligand-binding domains, DNA-binding domains, and N-terminal domains. There are four subtypes of TRs: TRα1, TRα2, TRβ1, and TRβ2. The heart primarily uses TRα1, while the liver primarily uses TRβ1. TRβ2 mRNA expression is mainly limited to the pituitary gland and hypothalamus. TRα1, TRβ1, and TRβ2 can bind to thyroid hormones and produce corresponding physiological effects. TRα2 does not bind to thyroid hormones.

[0006] Taking full advantage of thyroid hormones in increasing metabolic rate, oxygen consumption, and heat release can bring therapeutic benefits, such as in treating obesity. Hyperthyroidism is often accompanied by an overall increase in basal metabolic rate (BMR) along with food intake, and a weight loss of about 15%; while hypothyroidism is often accompanied by a weight gain of 25-30%. Most patients treated with T3 for hypothyroidism experience weight gain.

[0007] Furthermore, thyroid hormones can also lower serum low-density lipoprotein (LDL) (Journal of Molecular and Cellular Cardiology 37(2004):1137-1146). Existing studies have shown that hyperthyroidism can significantly reduce serum total cholesterol, mainly because thyroid hormones increase the expression of LDL receptors in the liver, thereby promoting the metabolism of cholesterol into bile acids; hypothyroidism is also associated with hypercholesterolemia. Therefore, thyroid hormones may reduce the incidence of atherosclerosis and other cardiovascular diseases.

[0008] Treatment with thyroid hormones often results in supraphysiological side effects due to individual variability, including cardiac problems (primarily tachycardia), muscle weakness, and excessive weight loss. Long-term use can also lead to bone loss. By modifying thyroid hormones to reduce the adverse effects of the thyroid hormones themselves while retaining their beneficial effects, suitable drugs can be developed to treat corresponding diseases: obesity, hyperlipidemia, hypercholesterolemia, diabetes, liver diseases (fatty liver, NASH, NAFLD, etc.), cardiovascular diseases (atherosclerosis, etc.), thyroid diseases (hypothyroidism, thyroid cancer, etc.), and other related conditions.

[0009] The publicly disclosed pyridazinone thyroid hormone analogues, such as CN101228135B, were applied for by Madrigal Pharmaceuticals. Its representative compound, MGL3196, is currently undergoing phase III clinical trials for NASH and NAFLD. Due to issues such as low activity and poor permeability, the oral dosage is 80-100 mg per day, which is significantly higher than other products targeting the same target.

[0010]

[0011] Another patent, CN1882327 C, filed by Viking Therapeutics, describes its compound VK2809, which is currently undergoing a Phase 2b clinical trial for NASH. Phase 1 clinical data indicates safety concerns, including elevated liver enzymes (liver damage) and a narrow therapeutic window. Furthermore, cartilage damage was observed in preclinical toxicology studies (J.Med.Chem.2014,57,3912-3923). Bristol-Myers Squibb Co.'s patent CN1216857C, describing its compound eprotirome, has terminated its Phase 3 clinical trial. Reported clinical data also show elevated liver enzymes, and cartilage damage was observed in preclinical toxicology studies. Other patents, such as those for pyridine derivatives (CN102459185) and indole derivatives (WO2002051805), have only reported activity data, with no further research or clinical trials reported. Summary of the Invention

[0012] In response to the problems existing in the reports, this application provides a new class of thyroid hormone β receptor agonists with better activity, selectivity or safety.

[0013] One aspect of this application is to provide a novel thyroid hormone β-receptor agonist compound as shown in Formula I, a pharmaceutically acceptable salt thereof, or a prodrug thereof:

[0014]

[0015] in,

[0016] R 1 The group can be hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclic, optionally substituted heteroaryl, optionally substituted amino, optionally substituted carbamoyl, or -COR. 10 ;

[0017] X is an optional substituted methylene, -O-, -S-, or -SO2-;

[0018] R a Selected from hydrogen, halogens, C 1-6 Straight-chain, branched, or cycloalkyl groups; or two adjacent R groups. a Bonding to form carbon rings or heterocycles;

[0019] L1 can be a single bond, methylene, -CH=CH-, -O-, -CO-, -NR3-, -NR3CO-, -CONR3-, -CH2NR3-, or -S-;

[0020] L2 is a single bond or -(CR4R5)p ;

[0021] R2 is a carboxyl group, or a group represented by the following formula:

[0022] R3 is hydrogen or an alkyl group that is optionally substituted;

[0023] R4 and R5 are each independently selected from hydrogen, halogen, or optionally substituted alkyl groups, or R4 and R5 are bonded together to form a cycloalkyl group;

[0024] R6 can be hydrogen, cyano, amino, COOH, or C. 1-6 Alkyl, C 1-6 Haloalkyl, C 3-6 cycloalkyl or C 3-6 Halogenated cycloalkyl groups;

[0025] R8 represents hydrogen, cyano, COOH, or C. 1-6 Alkyl, C 1-6 Haloalkyl, C 3-6 cycloalkyl or C 3-6 Halogenated cycloalkyl groups;

[0026] R7 and R9 are hydrogen and C, respectively. 1-3 Alkyl or C 1-3 Halogenated alkyl groups;

[0027] R 10 Optionally substituted alkyl, amino, hydroxyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclic, optionally substituted heteroaryl;

[0028] n can be 0, 1, 2, 3, or 4;

[0029] p is 0, 1, or 2.

[0030] In some preferred embodiments, R 1 For hydrogen or -COR 10 Or, one can choose to be hydrogen, deuterium, tritium, or C. 1-6 Alkyl, hydroxyl, halogen, CN-substituted alkyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclic, heteroaryl, amino, or carbamoyl. In some preferred embodiments, R 1 -COR 10 Or, optionally, hydrogen, deuterium, tritium, or C. 1-6 Alkyl, hydroxyl, halogen, CN-substituted C 1-10 Alkyl, C 3-10 cycloalkyl, C 3-10 cycloalkyl C 1-6 Alkyl, C 5-10 Aryl, C 5-10 Aryl C 1-6Alkyl, 5-10 membered heterocyclic, 5-10 membered heteroaryl, amino, or carbamoyl. In some preferred embodiments, R 1 -COR 10 Or, optionally, hydrogen, deuterium, tritium, or C. 1-6 Alkyl, hydroxyl, halogen, CN-substituted C 1-8 Alkyl, C 3-8 cycloalkyl, C 3-8 cycloalkyl C 1-6 Alkyl, C 5-10 Aryl, C 5-10 Aryl C 1-6 Alkyl, 5-10 membered heterocyclic, 5-10 membered heteroaryl.

[0031] In some respects, the compound of formula I provided in this application is shown as that of formula II:

[0032]

[0033] Among them, R b R c R d R e Hydrogen, deuterium, halogens, C 1-6 Straight-chain, branched, or cycloalkyl; or R b R c Bonded to form a 5- or 6-membered cycloalkyl group or a 5- or 6-membered non-aromatic heterocycle containing one or two heteroatoms selected from nitrogen, oxygen, and sulfur atoms; or R d R e Bonded to form a 5- or 6-membered cycloalkyl group or a 5- or 6-membered non-aromatic heterocycle containing one or two heteroatoms selected from nitrogen, oxygen and sulfur atoms;

[0034] Other substituents are defined as described in Formula I above.

[0035] In some respects, the compound of formula II provided in this application is:

[0036]

[0037] Where R1 is the C that can be substituted by any choice. 1-6 Straight-chain, branched alkyl or C 3-8 cycloalkyl;

[0038] X is O, S, or -CH2-;

[0039] R b R c R d R e Hydrogen, deuterium, halogens, C 1-6 Straight-chain, branched, or cycloalkyl, or Rb R c Bonded to form a 5- or 6-membered cycloalkyl group or a 5- or 6-membered non-aromatic heterocycle containing one or two heteroatoms selected from nitrogen, oxygen, and sulfur atoms; or R d R e Bonded to form a 5- or 6-membered cycloalkyl group or a 5- or 6-membered non-aromatic heterocycle containing one or two heteroatoms selected from nitrogen, oxygen and sulfur atoms;

[0040] L1 is a single bond, -NR3-, -O, or -S-;

[0041] L2 is a single bond or -CH2-;

[0042] R2 is selected from groups represented by the following formula:

[0043] R3 is hydrogen or an optional substituted C. 1-6 alkyl;

[0044] R6 can be hydrogen, cyano, amino, COOH, or C. 1-6 Alkyl, C 1-6 Haloalkyl, C 3-6 cycloalkyl or C 3-6 Halogenated cycloalkyl groups;

[0045] R8 represents hydrogen, cyano, COOH, or C. 1-6 Alkyl, C 1-6 Haloalkyl, C 3-6 cycloalkyl or C 3-6 Halogenated cycloalkyl groups;

[0046] R7 and R9 are hydrogen and C, respectively. 1-3 Alkyl or C 1-3 Halogenated alkyl groups.

[0047] In some respects, the compound of formula II provided in this application is:

[0048]

[0049] Where R1 is the C that can be substituted by any choice. 1-6 Straight-chain and branched alkyl groups;

[0050] R b R c R d R e Hydrogen, deuterium, halogens, C 1-6 Straight-chain, branched, or cycloalkyl, or R b R cBonded to form a 5- or 6-membered cycloalkyl group or a 5- or 6-membered non-aromatic heterocycle containing one or two heteroatoms selected from nitrogen, oxygen, and sulfur atoms; or R d R e Bonded to form a 5- or 6-membered cycloalkyl group or a 5- or 6-membered non-aromatic heterocycle containing one or two heteroatoms selected from nitrogen, oxygen and sulfur atoms;

[0051] X is O, S, or -CH2-;

[0052] L1 is a single bond, -O-, -S-, or -NH-;

[0053] L2 is a single bond;

[0054] R2 is selected from groups represented by the following formula:

[0055] R6 is hydrogen, cyano, or C. 1-6 Alkyl, C 1-6 Halogenated alkyl groups;

[0056] R8 represents hydrogen, cyano, and C. 1-6 Alkyl, C 1-6 Halogenated alkyl groups;

[0057] R7 and R9 are hydrogen and C, respectively. 1-3 Alkyl or C 1-3 Halogenated alkyl groups.

[0058] In some respects, the compound of formula II provided in this application is:

[0059]

[0060] Where R1 is any of the following: hydrogen, deuterium, tritium, or C. 1-6 Alkyl, hydroxyl, halogen, CN-substituted C 1-6 Straight-chain or branched alkyl, benzyl or C 5-6 Cycloalkylmethylene, more preferably isopropyl or benzyl;

[0061] R b and R d For halogens, R c and R e For hydrogen, R b and R d Chlorine is a further preferred option;

[0062] X is O, S, or -CH2-;

[0063] L1 is a single bond, -O, -S-, or -NH-;

[0064] L2 is a single bond or -CH2-;

[0065] R2 is selected from groups represented by the following formula:

[0066] R6, R7, R8, and R9 are hydrogen or carbon. 1-6 Alkyl or C 3-8 Cycloalkyl.

[0067] In one respect, the compound of formula I provided in this application is a compound of formula III:

[0068]

[0069] in,

[0070] R b R c Hydrogen, deuterium, halogens, C 1-6 Straight-chain, branched, or cycloalkyl groups;

[0071] A is O or methylene;

[0072] Other substituents are defined as described in Formula I.

[0073] In other preferred embodiments, in the compounds of formula I provided in this application, R1 is selected from:

[0074] 1) C can be arbitrarily substituted 1-6 Straight-chain or branched alkyl groups;

[0075] 2) Optional substitution of C 3-8 cycloalkyl;

[0076] 3) Optionally substituted C atoms containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur atoms. 3-8 Non-aromatic heterocyclic groups;

[0077] 4) Optionally substituted phenyl groups;

[0078] 5) Optionally substituted C atoms containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur atoms. 5-6 Mixed aromatic compounds.

[0079] In other preferred embodiments, in the compounds of formula I provided in this application, R1 is selected from -(CR) 11 R 12 ) m R 13 ;where R 11 R 12 C selected from hydrogen, deuterium, halogen, hydroxyl, amino, carboxyl, or optionally substituted C 1-4 Alkyl; R 13 Selected from:

[0080] 1) Hydrogen or deuterium;

[0081] 2) Halogens;

[0082] 3) Hydroxyl group;

[0083] 4) Amino;

[0084] 5) Carboxyl group;

[0085] 6) Optional substitution of C 1-4 Alkyl or C 1-4 Alkoxy;

[0086] 7) Optional substitution of C 3-8 cycloalkyl;

[0087] 8) Optionally substituted C atoms containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur atoms. 3-8 Non-aromatic heterocyclic groups;

[0088] 9) Optionally substituted phenyl groups;

[0089] 10) Optionally substituted C atoms containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur atoms. 5-6 Mixed aromatics;

[0090] m can be 0, 1, 2, or 3.

[0091] In other preferred embodiments, in the compounds of formula I provided in this application, R1 is selected from -COR 10 : where R 10 Selected from:

[0092] 1) Amino;

[0093] 2) Hydroxyl group;

[0094] 3) Optional substitution of C 1-4 Alkyl or C 1-4 Alkoxy;

[0095] 4) Optional substitution of C 3-8 cycloalkyl;

[0096] 5) Optionally substituted C atoms containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur atoms. 3-8 Non-aromatic heterocyclic groups;

[0097] 6) Optionally substituted phenyl groups;

[0098] 7) Optionally substituted C atoms containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur atoms. 5-6 Mixed aromatic compounds.

[0099] In some preferred embodiments, in the compound of formula I, R 1 For hydrogen, C 1-10 Alkyl (preferably C)1-5 Alkyl), C 3-10 cycloalkyl (preferably C) 3-8 cycloalkyl), C 3-10 cycloalkyl C 1-6 Alkyl (preferably C) 3-8 cycloalkyl C 1-4 Alkyl), C 5-10 Aryl (preferably C) 5-8 Aryl), C 5-10 Aryl C 1-6 Alkyl (preferably C) 5-8 Aryl C 1-4 Alkyl groups, 5-10 membered heterocyclic groups containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur atoms, 5-10 membered heteroaryl groups, amino groups, or -COR groups containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur atoms. 10 The C 1-10 Alkyl (preferably C) 1-5 Alkyl), C 3-10 cycloalkyl (preferably C) 3-8 cycloalkyl), C 3-10 cycloalkyl C 1-6 Alkyl (preferably C) 3-8 cycloalkyl C 1-4 Alkyl), C 5-10 Aryl (preferably C) 5-8 Aryl), C 5-10 Aryl C 1-6 Alkyl (preferably C) 5-8 Aryl C 1-4 Alkyl groups, 5-10 membered heterocyclic groups containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur atoms, 5-10 membered heteroaryl groups containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur atoms, or amino groups that are unsubstituted, or can be substituted by deuterium, tritium, or C. 1-6 Alkyl, hydroxyl, halogen, CN substitution;

[0100] X is methylene, -O-, -S-, or -SO2-;

[0101] R a Hydrogen, deuterium, halogens, C 1-6 Straight-chain, branched, or cycloalkyl groups; or two adjacent R groups. a Bonded to form 5-10 membered carbon rings, or 5-10 membered heterocycles containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur atoms;

[0102] L1 can be a single bond, methylene, -O-, -CO-, -NR3-, -NR3CO-, -CONR3-, -CH2NR3-, or -S-;

[0103] L2 is a single bond or C1-6 Alkyl (preferably C) 1-4 alkyl);

[0104] R2 is a carboxyl group, or a group represented by the following formula:

[0105] R3 is hydrogen or C. 1-6 alkyl;

[0106] R6 can be hydrogen, cyano, amino, COOH, or C. 1-6 Alkyl or C 1-6 Halogenated alkyl groups;

[0107] R8 represents hydrogen, cyano, COOH, or C. 1-6 Alkyl or C 1-6 Halogenated alkyl groups;

[0108] R7 and R9 are hydrogen and C, respectively. 1-3 Alkyl or C 1-3 Halogenated alkyl groups;

[0109] R 10 C 3-10 cycloalkyl (preferably C) 3-8 cycloalkyl), C 5-10 Aryl (preferably C) 5-8 aryl), 5-10 membered heterocyclic groups containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur atoms; 5-10 membered heteroaryl groups containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur atoms;

[0110] n can be 0, 1, 2, 3, or 4.

[0111] In some preferred embodiments, in the compound of formula I provided in this application, R 1 C 1-8 Alkyl (preferably C) 1-5 Alkyl), C 3-8 cycloalkyl (preferably C) 3-6 cycloalkyl), C 3-8 cycloalkyl C 1-5 Alkyl (preferably C) 3-6 cycloalkyl C 1-3 Alkyl), C 5-8 Aryl (preferably C) 5-6 Aryl), C 5-8 Aryl C 1-5 Alkyl (preferably C) 5-6 Aryl C 1-3 Alkyl groups, 5-8 membered heterocyclic groups containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur atoms, 5-8 membered heteroaryl groups, amino groups, or -COR groups containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur atoms. 10 The C1-8 Alkyl (preferably C) 1-5 Alkyl), C 3-8 cycloalkyl (preferably C) 3-6 cycloalkyl), C 3-8 cycloalkyl C 1-5 Alkyl (preferably C) 3-6 cycloalkyl C 1-3 Alkyl), C 5-8 Aryl (preferably C) 5-6 Aryl), C 5-8 Aryl C 1-5 Alkyl (preferably C) 5-6 Aryl C 1-3 Alkyl groups, 5-8 membered heterocyclic groups containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur atoms; 5-8 membered heteroaryl groups containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur atoms; or amino groups that are unsubstituted, or can be substituted by deuterium, tritium, or C. 1-6 Alkyl, hydroxyl, halogen, CN substitution;

[0112] X is methylene, -O-, -S-, or -SO2-;

[0113] R a Halogen, C 1-4 Straight-chain or branched alkyl groups; or two adjacent R groups. a Bonded to form 5-7 membered carbon rings, or 5-7 membered heterocycles containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur atoms;

[0114] L1 can be a single bond, -O-, -NR3-, -NR3CO-, -CONR3-, -CH2NR3-, or -S-;

[0115] L2 is a single bond or C 1-5 Alkyl (preferably C) 1-3 alkyl);

[0116] R2 is a carboxyl group, or a group represented by the following formula:

[0117] R3 is hydrogen or C. 1-3 alkyl;

[0118] R6 is hydrogen, cyano, COOH, or C. 1-4 alkyl;

[0119] R8 is hydrogen or C. 1-4 alkyl;

[0120] R7 and R9 are hydrogen or C. 1-3 alkyl;

[0121] R 10 C 5-8 Aryl (preferably C)5-6 aryl), or a 5-8 membered heteroaryl (preferably 5-6 membered heteroaryl) containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur atoms;

[0122] n is 1, 2, or 3.

[0123] In some preferred embodiments, in the compound of formula I, R 1 It can be methyl, ethyl, propyl, butyl, pentyl, cyclopropane, cyclobutane, cyclopentane, cyclohexane, cyclopropanemethyl, cyclobutanemethyl, cyclopentanemethyl, cyclohexanemethyl, phenyl, benzyl, or -COR. 10 The methyl, ethyl, propyl, butyl, pentyl, cyclopropane, cyclobutane, cyclopentane, cyclohexane, cyclopropanemethyl, cyclobutanemethyl, cyclopentanemethyl, cyclohexanemethyl, phenyl, and benzyl groups are unsubstituted or can be substituted by deuterium or C. 1-3 Alkyl, hydroxyl, halogen, CN substitution;

[0124] X is methylene, -O-, -S-, or -SO2-;

[0125] R a It is a halogen; or two adjacent R a Bonded to form a 5-membered carbon ring, or a 5-membered heterocycle containing 1-2 heteroatoms selected from nitrogen, oxygen and sulfur atoms;

[0126] L1 can be a single bond, -O-, -NH-, -NHCO-, -CONH-, -CH2NH-, or -S-;

[0127] L2 is a single bond, methyl, ethyl, or propyl;

[0128] R2 is a carboxyl group, or a group represented by the following formula:

[0129] R6 is hydrogen, cyano, COOH, methyl, ethyl, or propyl;

[0130] R8 is hydrogen, methyl, ethyl, or propyl;

[0131] R7 and R9 are hydrogen or methyl;

[0132] R 10 It is phenyl;

[0133] n is 2 or 3.

[0134] In some preferred embodiments, in the compound of formula I, R 1The methyl, ethyl, propyl, butyl, pentyl, cyclopropane, cyclobutane, cyclopentane, cyclohexane, cyclopropane methyl, cyclobutane methyl, cyclopentane methyl, cyclohexane methyl, phenyl, or benzyl groups are used, wherein the methyl, ethyl, propyl, butyl, pentyl, cyclopropane, cyclobutane, cyclopentane, cyclohexane methyl, cyclopropane methyl, cyclobutane methyl, cyclopentane methyl, cyclohexane methyl, phenyl, or benzyl groups are unsubstituted or can be substituted by deuterium or C. 1-3 Alkyl, hydroxyl, F, Cl, Br, CN substitution;

[0135] X is methylene, -O-, or -S-;

[0136] R a For F, Cl, Br; or two adjacent R a Bonded to form a 5-membered carbon ring, or a 5-membered heterocycle containing 1-2 heteroatoms selected from nitrogen, oxygen and sulfur atoms;

[0137] L1 is a single bond, -O-, -NH-, or -NHCO-;

[0138] L2 is a single bond, methyl, ethyl, or propyl;

[0139] R2 is a carboxyl group, or a group represented by the following formula:

[0140] R6 is hydrogen, cyano, or methyl;

[0141] R7 is hydrogen;

[0142] n is 2 or 3.

[0143] In some specific embodiments, in the compounds of formula I provided in this application, R1 is selected from:

[0144] In certain specific embodiments, in the compound of formula I provided in this application, R b R c R d R e Selected from hydrogen, deuterium, or halogens.

[0145] In some specific embodiments, in the compounds of Formula I provided in this application, L1 is selected from single bonds, -O-, -NH-, -NHCO-, and -NHCH2-.

[0146] In some specific embodiments, in the Formula I compounds provided in this application, L2 is selected from single bonds or methylene (-CH2-).

[0147] In some specific embodiments, in the compound of formula I provided in this application, R2 is selected from: carboxyl group,

[0148] In some specific embodiments, the compounds of formula I provided in this application are selected from:

[0149]

[0150]

[0151] Another aspect of this application is to provide a pharmaceutical composition comprising a compound of Formula I of this application, a pharmaceutically acceptable salt thereof or a prodrug thereof, and one or more pharmaceutically acceptable carriers.

[0152] Another aspect of this application is to provide the use of the compound of this application, its pharmaceutically acceptable salt, or its prodrug in the preparation of a medicament for the prevention or treatment of diseases related to the action of β-receptor agonists (e.g., obesity, hyperlipidemia, hypercholesterolemia, diabetes, steatohepatitis, non-alcoholic steatohepatitis, non-alcoholic fatty liver disease, atherosclerosis, thyroid cancer, hypothyroidism, or related conditions). Alternatively, this application provides the aforementioned compound, its pharmaceutically acceptable salt, or its prodrug for the prevention or treatment of diseases related to the action of β-receptor agonists. Alternatively, this application provides a method for the prevention or treatment of diseases related to the action of β-receptor agonists, comprising administering the aforementioned compound, its pharmaceutically acceptable salt, or its prodrug to a subject in need. Preferably, the diseases associated with the action of the β-receptor agonist include, but are not limited to: hypercholesterolemia, hyperlipidemia, hypertriglyceridemia, familial hypercholesterolemia, dyslipidemia, thyroid cancer, hypothyroidism, potential hypothyroidism, atherosclerosis, metabolic syndrome, obesity, diabetes, cardiovascular disease, coronary artery disease, myocardial infarction, ventricular dysfunction, heart failure, fatty liver, cirrhosis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), depression, dementia, osteoporosis, alopecia, nail diseases, skin diseases, kidney diseases, chronic renal failure, and / or cancer, especially hypercholesterolemia, hyperlipidemia, hypertriglyceridemia, familial hypercholesterolemia, dyslipidemia, atherosclerosis, hypothyroidism, and / or potential hypothyroidism.

[0153] definition

[0154] Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art. References to technical terms herein refer to techniques commonly understood in the art, including variations or equivalent substitutions of techniques that are obvious to one of ordinary skill in the art. While it is believed that the following terms will be well understood by one of ordinary skill in the art, the following definitions are set forth to better explain this application.

[0155] As used herein, the terms “including,” “comprising,” “having,” “containing,” or “involving,” and their other variations herein, are inclusive or open-ended and do not exclude other unlisted elements or method steps.

[0156] As used herein, the term “hydrogen” and the hydrogen in each group encompass its naturally occurring isotopes protium (P), deuterium (D) or tritium (T).

[0157] Alkyl groups are a class of straight-chain or linear organic groups containing only carbon and hydrogen atoms. Examples of alkyl groups include C... 1-10 C is preferred 1-6 C, more preferably 1-4 Straight-chain or branched alkyl groups, such as C1, C2, C3, C4, C5, C6, C7, C8, C9 or C 10 Alkyl groups, specifically, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, 1-methylpropyl, pentyl, and hexyl.

[0158] "Halogens" include fluorine, chlorine, bromine, and iodine atoms.

[0159] "Cycloalkyl" includes C 3-14 C is preferred 3-10 C, more preferably 6-10 The monocyclic, bicyclic, or tricyclic nonaromatic carbon rings, optionally partially or fully saturated.

[0160] "Heterocyclic group" includes monocyclic, bicyclic, or tricyclic non-aromatic carbocyclic or cycloalkanes containing one or more (e.g., 1-5, 1-4, 1-3, or 1-2) heteroatoms selected from phosphorus, nitrogen, oxygen, and sulfur atoms (particularly nitrogen, oxygen, and sulfur atoms); exemplary, "heterocyclic" includes 5- to 12-membered monocyclic or bicyclic non-aromatic carbocyclic or cycloalkanes containing 1-4 heteroatoms selected from nitrogen, oxygen, and sulfur atoms, optionally partially or fully saturated.

[0161] "Aryl" refers to a monocyclic or polycyclic aromatic group consisting entirely of carbon atoms and having a conjugated π-electron system. For example, as used herein, the term "C" refers to a monocyclic or polycyclic aromatic group consisting entirely of carbon atoms and having a conjugated π-electron system. 6-14"Aryl" refers to an aromatic group containing 6 to 14 carbon atoms, such as phenyl or naphthyl. The aryl group is optionally substituented by one or more (such as 1 to 3) suitable substituents (e.g., halogen, -OH, -CN, -NO2, C). 1-6 Alkyl groups, etc., are substituted.

[0162] "Heteroaryl" is an aromatic cyclic group containing at least one heteroatom (nitrogen, oxygen, or sulfur) and a carbon atom, including 5- or 6-membered monocyclic compounds, 8- to 10-membered bicyclic groups wherein the same or different monocyclic heteroaryl rings are fused together, and 8- to 10-membered bicyclic groups wherein the monocyclic heteroaryl rings are fused with benzene. Specific examples of heteroaryl groups include furanyl, thiophene, pyrrole, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, thiazolyl, furazonyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, indolyl, inzolyl, benzimidazolyl, purinyl, quinolinyl, isoquinolinyl, diazonaphthyl, quinoxolinyl, quinazolinyl, zolinyl, benzofuranyl, benzothiophene, benzooxazolyl, benzothiazolyl, benzoisooxazolyl, and benzoisothiazolyl, etc.

[0163] As used herein, the term "substitution" refers to the selective replacement of one or more (e.g., one, two, three, or four) hydrogen atoms on a specified atom by a designated group, provided that the substitution does not exceed the normal valence of the specified atom in the present case and that the substitution forms a stable compound. Combinations of substituents and / or variables are permitted only if such combinations form a stable compound.

[0164] If a substituent is described as “optionally substituted,” then the substituent may be (1) unsubstituted or (2) substituted. If the carbon of the substituent is described as being optionally substituted with one or more of the substituents in the list, then one or more hydrogens on the carbon (to the extent that any hydrogens are present) may be substituted individually and / or together with independently selected substituents. If the nitrogen of the substituent is described as being optionally substituted with one or more of the substituents in the list, then one or more hydrogens on the nitrogen (to the extent that any hydrogens are present) may be substituted individually with independently selected substituents.

[0165] "Optional substitution" can be achieved by 1-5, preferably 1-3, substituents, and said substituents include (1) alkyl groups substituted with 1-3 groups selected from halogen, hydroxyl, carboxyl, amino, aryl, heteroaryl, cycloalkyl, and heterocyclic groups; (2) carbocyclic groups substituted with 1-3 groups selected from alkyl, halogen, hydroxyl, carboxyl, haloalkyl, alkoxy, haloalkoxy, alkanoyl, and cyano groups; and (3) groups substituted with 1-3 groups selected from alkyl, halogen, hydroxyl, carboxyl, haloalkyl, alkoxy, haloalkoxy, alkyl, cyanoyl, alkyl, hydroxyl, carboxyl, haloalkyl, alkoxy, haloalkoxy, alkylyl, hydroxyl, carboxyl, alkylyl, alkoxy, alkylyl ... (4) A heterocyclic group substituted with a group selected from alkyl, halogen, hydroxyl, carboxyl, haloalkyl, alkoxy, haloalkoxy, alkylyl and cyano; (5) A heteroaryl group substituted with a group selected from alkyl, halogen, hydroxyl, carboxyl, haloalkyl, alkoxy, haloalkoxy, alkylyl and cyano; (6) hydroxyl; (7) alkoxy; (8) halogen; (9) an amino group optionally substituted with one or two alkyl groups; and (10) oxy.

[0166] This application also includes all pharmaceutically acceptable isotopically labeled compounds identical to those of this application, except that one or more atoms are replaced by atoms having the same atomic number but a different atomic mass or mass number than the dominant atomic mass or mass number in nature. Examples of isotopes suitable for inclusion in the compounds of this application include (but are not limited to) isotopes of hydrogen (e.g., deuterium). 2 H), tritium ( 3 H); carbon isotopes (e.g., H); 11 C 13 C and 14 C); isotopes of chlorine (e.g.) 36 Cl); isotopes of fluorine (e.g., Cl); 18 F); isotopes of iodine (e.g., F); 123 I and 125 I); nitrogen isotopes (e.g.) 13 N and 15 N); isotopes of oxygen (e.g., N); 15 O、 17 O and 18 O); isotopes of phosphorus (e.g., O); phosphorus isotopes (e.g., O); 32 P); and isotopes of sulfur (e.g. 35 S). Certain isotope-labeled compounds of this application (e.g., those doped with radioactive isotopes) can be used in drug and / or substrate tissue distribution studies (e.g., analysis). Radioactive isotope tritium (i.e. 3 H) and carbon-14 (i.e. 14 C) It is particularly suitable for this purpose due to its ease of incorporation and detection. Using positron-emitting isotopes (e.g.) 11 C 18 F, 15 O and 13Substitution of N) can be used to examine substrate receptor occupancy in positron emission tomography (PET) studies.

[0167] The structures described herein also include all isomers (e.g., enantiomers, diastereomers, and geometric isomers (or conformational isomers) of that structure; for example, R and S configurations for each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers. Therefore, single stereochemical isomers of these compounds, as well as mixtures of enantiomers, diastereomers, and geometric isomers (or conformational isomers), are within the scope of this application. Unless otherwise specified, all tautomers of the compounds described herein are within the scope of this application. Furthermore, unless otherwise specified, the structures described herein also include compounds that differ only in the presence of one or more isotopically enriched atoms.

[0168] This document covers all possible crystalline forms or polymorphs of the compounds of this application, which may be a single crystalline form or a mixture of more than one polymorph in any proportion.

[0169] It should also be understood that certain compounds of this application may exist in their free form for therapeutic purposes, or, where appropriate, in their pharmaceutically acceptable derivative forms. In this application, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, solvates, N-oxides, metabolites, or prodrugs, which, upon administration to a patient in need, can directly or indirectly provide the compound of this application or its metabolites or residues. Therefore, when referring to "compounds of this application" herein, it is also intended to cover the various derivative forms of the compounds described above.

[0170] Pharmaceutically acceptable salts of the compounds described in this application include their acid addition salts and base addition salts, and the types of salts are not particularly limited, as long as they are physiologically acceptable. Suitable examples of pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochlorides, hydrobroms, sulfates, nitrates, phosphates, acetates, trifluoroacetates, tartrates, fumarates, oxalates, maleates, citrates, succinates, methanesulfonates, benzenesulfonates, malates, aspartates, glucoheponicates, glucuronides, orotates, palmitates, and other similar salts. Suitable examples of pharmaceutically acceptable base addition salts include, but are not limited to, sodium salts, potassium salts, ammonium salts, calcium salts, magnesium salts, aluminum salts, iron salts, histidine salts, arginine salts, choline salts, and other similar salts.

[0171] The compounds of this application may exist in the form of solvates (preferably hydrates), wherein the compounds of this application contain a polar solvent, particularly such as water, methanol, or ethanol, as a structural element of the compound's crystal lattice. The amount of the polar solvent, particularly water, may be stoichiometric or non-stoichiometric.

[0172] Those skilled in the art will understand that not all nitrogen-containing heterocycles can form N-oxides because nitrogen requires available lone pairs of electrons to be oxidized into oxides; those skilled in the art will identify nitrogen-containing heterocycles that can form N-oxides. Those skilled in the art will also recognize that tertiary amines can form N-oxides. Synthetic methods for preparing N-oxides of heterocycles and tertiary amines are well known to those skilled in the art, including the oxidation of heterocycles and tertiary amines with peroxy acids such as peracetic acid and m-chloroperoxybenzoic acid (MCPBA), hydrogen peroxide, alkyl peroxides such as tert-butyl peroxide, sodium perborate, and dioxiranes such as dimethyldioxirane.

[0173] The scope of this application also includes metabolites of the compounds of this application, i.e., substances formed in the body when the compounds of this application are administered. Such products can be generated by, for example, oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc., of the administered compound. Therefore, this application includes metabolites of the compounds of this application, including compounds prepared by methods that expose the compounds of this application to mammals for a time sufficient to produce their metabolites.

[0174] This application further includes, within its scope, prodrugs of the compounds of this application, which are certain derivatives of the compounds of this application that may have little or no pharmacological activity on their own, which, when administered to or onto the body, can be converted, for example, by hydrolysis and cleavage into the compounds of this application having the desired activity. Typically, such prodrugs are functional group derivatives of the compounds that are readily converted in vivo into the compounds with the desired therapeutic activity.

[0175] In this application, "pharmaceutically acceptable carrier" refers to a pharmacologically and pharmaceutically acceptable additive that is administered together with the active ingredient, and may include excipients, disintegrants, binders, lubricants, coating agents, dyes, diluents, bases, and isotropic agents, etc.

[0176] The dosage forms include, but are not limited to, tablets, capsules, lozenges, hard candies, powders, sprays, creams, ointments, drops, suppositories, gels, pastes, lotions, aqueous suspensions, injectable solutions, elixirs, and syrups.

[0177] Examples of dosage forms suitable for oral administration include tablets, capsules, powders, fine granules, granules, liquids, and syrups. Examples of dosage forms suitable for non-oral administration include injections, drops, and suppositories.

[0178] In this document, unless otherwise stated, singular terms cover the plural referents, and vice versa.

[0179] In this document, unless otherwise stated, the term “subject” may be used interchangeably with “individual” or “patient”, including vertebrates such as birds, fish, mammals such as, but not limited to, mice, rats, guinea pigs, dogs, pigs, sheep, cattle, chickens, rabbits, monkeys (e.g., rhesus monkeys), humans, etc.

[0180] In this document, unless otherwise stated, all figures representing amounts of components, measurements, or reaction conditions as used herein should be understood to be modified in all cases by the term "about" to indicate possible measurement error. For example, when associated with a percentage, the term "about" may mean ±1%.

[0181] The compounds of Formula I of this application exhibit thyroid hormone β-receptor agonist action and are capable of preventing or treating diseases related to β-receptor agonist action, such as drugs for the prevention, reduction, and / or treatment of the following diseases: hypercholesterolemia, hyperlipidemia, hypertriglyceridemia, familial hypercholesterolemia, dyslipidemia, thyroid cancer, hypothyroidism, potential hypothyroidism, atherosclerosis, metabolic syndrome, obesity, diabetes, cardiovascular disease, coronary artery disease, myocardial infarction, ventricular dysfunction, heart failure, fatty liver, cirrhosis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), depression, dementia, osteoporosis, alopecia, nail diseases, skin diseases, kidney diseases, chronic renal failure, and / or cancer, especially hypercholesterolemia, hyperlipidemia, hypertriglyceridemia, familial hypercholesterolemia, dyslipidemia, atherosclerosis, hypothyroidism, and / or potential hypothyroidism. Attached Figure Description

[0182] Appendix Figure 1 This is the result of the fibrosis evaluation.

[0183] Appendix Figure 2 The NAS rating results. Detailed Implementation

[0184] To make the objectives and technical solutions of this application clearer, the following detailed embodiments further illustrate this application. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of this application. Furthermore, any specific experimental methods not mentioned in the following embodiments are performed according to conventional experimental methods.

[0185] Example 1: Synthesis of the key intermediate KH01

[0186]

[0187] Compound KH01-1: At 0°C, the starting materials 2-chloro-5-bromopyrimidine (KH01-1a) (100 g, 516 mmol, 1.00 eq), isobutyric acid (1b) (36.4 g, 413 mmol, 38.3 mL, 0.80 eq), potassium persulfate (111 g, 413 mmol, 82.8 mL, 0.80 eq), and silver nitrate (17.5 g, 103 mmol, 0.20 eq) were added to a round-bottom flask, along with 1 L of dichloromethane and 1 L of water. After stirring until homogeneous, some solids remained undissolved. The mixture was then cooled to room temperature (25°C) and reacted under nitrogen protection for 12 h with stirring. TLC analysis showed that the starting materials reacted completely, and new spots were observed. The reaction solution was filtered, and the filter cake was washed twice with dichloromethane (1 L each time). The filtrate was collected and concentrated under reduced pressure. Purification was achieved by silica gel (100-200 mesh) column chromatography (petroleum ether / ethyl acetate 100:1) to give 54.0 g of oily substance KH01-1, yield: 44.3%. LCMS:MS (ESI) m / z = 236.9 [M+H] + . 1 H NMR (400MHz CDC13): δ8.58 (s, 1H), 3.48 -3.41 (m, 1H), 1.29 (d, J = 6.8Hz, 6H).

[0188] Compound KH01-2: KH01-1 (15.0 g, 63.6 mmol, 1.00 eq) was added to a round-bottom flask, followed by compound 1a (11.3 g, 63.6 mmol, 1.00 eq), cesium carbonate (62.2 g, 191 mmol, 3.00 eq), and N,N-dimethylformamide (150 mL). After thorough stirring, the mixture was purged with nitrogen three times and reacted at 80 °C for 2 h under nitrogen protection. TLC analysis confirmed complete reaction of the starting materials. The reaction solution was poured into 100 mL of water, stirred, and the solid dissolved. Ethyl acetate was added for extraction (100 mL * 2 times). The combined ethyl acetate layers were washed with saturated brine (100 mL * 2 times), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel (100-200 mesh) column chromatography (petroleum ether / ethyl acetate 1:2) to obtain 13.0 g of yellow solid KH01-2, yield: 54.1%. LCMS:MS(ESI)m / z=378.0[M+H] + . 1 H NMR (400MHz CDC13): δ8.46 (s, 1H), 6.68 (s, 2H), 3.78 (s, 2H), 3.43-3.36 (m, 1H), 1.22 (d, J = 6.8Hz, 6H).

[0189] Compound KH01-3: Starting material KH01-2 (10.0 g, 26.5 mmol, 1.00 eq), bis-pinacolborate (pin2B2) (13.4 g, 53.0 mmol, 2.00 eq), 1,1-bis(diphenylphosphine)ferrocene palladium dichloromethane complex (1.08 g, 1.33 mmol, 0.05 eq), and potassium acetate (5.21 g, 53.0 mmol, 2.00 eq) were dissolved in dioxane (100 mL). The mixture was purged three times with nitrogen and reacted at 110 °C for 4 h under nitrogen protection. TLC analysis confirmed complete reaction of the starting material. 200 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (200 mL * 3). The ethyl acetate layers were combined, washed with saturated brine (300 mL * 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain 12.0 g of a brown oily substance. No purification was required; the mixture was directly used in the next reaction step. LCMS:MS(ESI)m / z=425.2[M+H] + .

[0190] Compound KH01: At an external temperature of 0℃, starting material KH01-3 (12.0 g, 28.2 mmol, 1.00 eq) and 30.0% hydrogen peroxide (6.74 g, 59.4 mmol, 5.71 mL, 2.10 eq) were dissolved in tetrahydrofuran (120 mL). Under nitrogen protection, the mixture was stirred at room temperature (25℃) for 2 h. After the reaction was confirmed to be complete by TLC, 50 mL of 2M sodium sulfite solution was added to terminate the reaction. The mixture was extracted with dichloromethane (5 mL * 3), and the organic layer was collected, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel (100-200 mesh) column chromatography (petroleum ether / ethyl acetate 1:2) to give 4.80 g of yellow solid, yield: 53.4%. LCMS:MS(ESI) m / z = 314.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ9.85 (br s, 1H), 7.95 (s, 1H), 6.66 (s, 2H), 5.52 (s, 2H), 3.31-3.18 (m, 1H), 1.10 (d, J = 6.8Hz, 6H).

[0191] Example 2: Synthesis of compound KH02

[0192]

[0193] Compound KH02-1: Compound KH01 (0.152 g, 0.485 mmol), diethyl phosphite (0.104 g, 0.754 mmol), paraformaldehyde (0.095 g, 1.055 mmol), and sodium sulfate (0.156 g, 1.098 mmol) were weighed and placed in a single-necked flask. Toluene (8 mL) was added to partially dissolve the compound, and the mixture was kept under N2 protection. After reacting at 110 °C for 3 hours, the reaction was monitored by TLC until the reactants were completely reacted and a major polarity point was formed. The reaction was then stopped. 100 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 mL * 3). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1:2) to obtain a colorless oily substance KH02-1 (142 mg, 63%). LC-MS (ESI, m / z): 465.3 [M+H] + .

[0194] Compound KH02: Compound KH02-1 (0.142 g, 0.306 mmol) was placed in a single-necked flask and dissolved in dichloromethane (10 mL) under N2 protection. Trimethylbromosilane (3.2 mL) was slowly added dropwise to the reaction system at an external temperature of -10 °C. After maintaining this temperature for 30 min, the reaction was slowly allowed to proceed to room temperature overnight. LCMS was used to confirm complete reaction of the starting material. The reaction solution was directly evaporated to dryness, and the product was purified by preparative HPLC. The target product was collected and lyophilized to obtain a pale yellow solid compound KH02 (55 mg, 44%). 1 H NMR (400MHz, DMSO): δ9.87 (s, 1H), 7.95 (s, 1H), 6.82 (s, 2H), 4.61-5.22 (m, 3H), 3.27 (m, 3H), 1.12 (d, J = 4MHz, 6H). LC-MS (ESI, m / z): 408.0 [M+1] + .

[0195] Example 3: Synthesis of compound KH03

[0196]

[0197] Compound KH03-1: Compound KH01 (0.265 g, 0.846 mmol) was added to 11.2 mL of water. 5.6 mL of concentrated hydrochloric acid was added at 0°C. Sodium nitrite (0.072 g, 1.043 mmol) was dissolved in 0.8 mL of water and slowly added dropwise to the above reaction solution. The mixture was stirred at 0°C for 1.5 h to form a solution. Separately, compound 3a (0.148 g, 0.948 mmol) was dissolved in 19.4 mL of water. 5.6 mL of pyridine was added at 0°C and stirred at this temperature for 1.5 h. Then, the solution was rapidly added to the above reaction mixture at 0°C, resulting in the formation of an orange-red solid. The reaction solution was slowly raised to room temperature (25°C) and the reaction continued overnight. TLC monitoring showed that the reaction was complete. The solid was filtered directly using a filter funnel and washed three times each with 50 mL of water and PE. A bright orange-red solid, KH03-1 (380 mg, 93.8%), was collected. LC-MS (ESI, m / z): 482.3 [M+1] + .

[0198] Compound KH03: Compound KH03-1 (0.380 g, 0.931 mmol) and sodium acetate (0.650 g, 7.926 mmol) were placed in a single-necked flask, dissolved in acetic acid (10 mL), and protected with N2. The reaction was carried out at 120 °C for 3 h. The reaction was stopped after TLC detection showed complete reaction of the starting materials. The reaction solution was cooled to 0 °C, and 100 mL of water was added. A large amount of solid precipitated. The solid was filtered directly through a filter funnel and washed three times with 50 mL of water and PE, respectively. Orange-red solid KH03 (230 mg, 66.9%) was collected. 1 HNMR (400MHz, DMSO) δ10.09 (s, 1H), 8.01 (s, 1H), 7.75 (s, 2H), 3.27-3.28 (m, 1H), 1.13 (d, J = 4MHz, 6H). LC-MS (ESI, m / z): 435.2[M+1] + .

[0199] Example 4: Synthesis of compound KH04

[0200]

[0201] Compound KH04-1: Compound KH01 (0.0512 g, 0.1635 mmol), ethyl glyoxylate (0.0274 g, 0.268 mmol), and sodium triacetoxyborohydride (0.1023 g, 0.483 mmol) were weighed and placed in a single-necked flask. 1,2-Dichloroethane (3 mL) was added to dissolve the compounds. The reaction was carried out at 75 °C for 3 hours. TLC monitoring showed complete reaction of the starting materials, with the formation of a major polarity point. The reaction was then stopped. 50 mL of dichloromethane and 100 mL of water were added to the reaction solution. After stirring for 10 min, the organic phase was separated. The aqueous phase was then extracted with dichloromethane (50 mL * 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by chromatography (petroleum ether / ethyl acetate = 1:2) to obtain a colorless oily substance KH04-1 (50 mg, 78.5%). LC-MS (ESI, m / z): 401.3 [M+1] + .

[0202] Compound KH04: Compound KH04-1 (50 mg, 0.125 mmol) and lithium hydroxide (35 mg, 1.458 mmol) were placed in a single-necked flask and dissolved in tetrahydrofuran / methanol / water (4:1:1, 6 mL). The reaction was allowed to proceed overnight at room temperature. The reaction was stopped after TLC detection to ensure complete reaction of the starting material. The reaction solution was diluted with 20 mL of water, the organic solvent was removed under reduced pressure, and the pH was adjusted to 3-4 at 0 °C. Dichloromethane (50 mL * 3) was added to the reaction solution for extraction. The organic phase was collected, dried over anhydrous Na2SO4, filtered and concentrated to remove the solvent, and the residue was purified by chromatography (dichloromethane / methanol = 5:1). The target product was collected, lyophilized, and yielded a white solid KH04 (24 mg, 51.6%). 1 H NMR(400MHz,DMSO)δ10.48(s,1H),8.00(s,1H),6.65(s,2H),5.98(s,1H),4.21-4.22(m, 1H),3.49(s,1H),3.34-3.19(m,2H),1.12(d,J=4MHz,6H).LC-MS(ESI,m / z):372.1[M+1] + .

[0203] Example 5: Synthesis of compound KH05

[0204]

[0205] 100 mg of compound KH03 was dissolved in 5 mL of acetic acid, and then 1 mL of concentrated hydrochloric acid was added. The mixture was stirred at 90 °C for 4 h. TLC analysis showed that the reaction proceeds were complete. The reaction solution was evaporated to dryness under reduced pressure, and the pH was adjusted to 9–10 with saturated sodium carbonate solution. After extraction with 50 mL of ethyl acetate, the organic phase was discarded. The aqueous phase was adjusted to 3–4, and the solution was extracted again with ethyl acetate (50 mL * 3). The organic phases were combined, dried over anhydrous sodium sulfate, and 70 mg of concentrated white solid was obtained. Yield: 67.1%. LC-MS (ESI, m / z): 455.3 [M+1] + .

[0206] Example 6: Synthesis of compound KH06

[0207]

[0208] 60 mg of KH05 was dissolved in 4 mL of mercaptoacetic acid. Under nitrogen protection, the mixture was stirred at 120 °C for 6 h. TLC analysis revealed the formation of a low-polarity product spot. The reaction was quenched by adding saturated sodium thiosulfate, and the mixture was extracted with ethyl acetate (25 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The solution was purified by chromatography (dichloromethane / methanol 10:1). The target product was collected and lyophilized to give approximately 15 mg of white solid KH06. Yield: 27.8%. LC-MS (ESI, m / z): 410.1 [M+1] + . 1 H NMR (400MHz, DMSO) δ: 12.49 (s, 1H), 10.08 (s, 1H), 8.01 (s, 2H), 7.71 (d, J = 4MHz, 1H), 3.27-3.33 (m, 1H), 1.12 (d, J = 4MHz, 6H).

[0209] Example 7: Synthesis of the key intermediate KH07-10

[0210]

[0211] Compound KH07-2: Starting material KH07-1 (25.0 g, 184 mmol, 1 eq) was dissolved in DMF (200 mL). NBS (32.9 g, 184 mmol, 1 eq) was slowly added at 0 °C. After the addition was complete, the mixture was stirred at 25 °C for 5 h. LCMS monitoring showed that the reaction was complete and a new spot (RT = 0.483) was observed. TLC (petroleum ether / ethyl acetate = 3 / 1) monitoring revealed two new spots. The reaction solution was diluted with water (250 mL) and extracted with ethyl acetate (250 mL * 2). The organic phases were combined, washed with saturated brine (250 mL * 2), dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 3 / 1) to give yellow solid KH07-2 (29.1 g, 135 mmol, yield 73.5%). MS(ESI) m / z: 216.1 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ 6.75-6.69 (m, 1H), 6.43 (d, J = 8.4Hz, 1H), 4.76 (s, 2H), 4.58-4.49 (m, 2H), 3.15-3.08 (m, 2H).

[0212] Compound KH07-3: Compound KH07-2 (28.0 g, 130 mmol, 1 eq) and TFAA (32.9 g, 156 mmol, 21.8 mL, 1.2 eq) were dissolved in dichloromethane (280 mL). DIEA (33.8 g, 261 mmol, 45.5 mL, 2 eq) was slowly added dropwise at 0 °C. After the addition was complete, the mixture was stirred at 25 °C for 1 h. The reaction mixture was monitored by TLC (petroleum ether / ethyl acetate = 5 / 1) to ensure complete reaction. The reaction mixture was poured into water (280 mL) and extracted with dichloromethane (300 mL * 3). The organic layers were combined, washed with saturated brine (280 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 3 / 1) to give a yellow solid KH07-3 (27.8 g, yield 68.5%). MS(ESI) m / z: 309.9 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ 11.26-10.86 (m, 1H), 7.15-7.10 (m, 1H), 7.09-7.03 (m, 1H), 4.71-4.62 (m, 1H), 4.66 (t, J = 8.8Hz, 1H), 3.25 (t, J = 8.8Hz, 2H).

[0213] Compound KH07-4: Compound KH07-3 (27.0 g, 87.0 mmol, 1 eq), Pd(dppf)Cl2·CH2Cl2 (3.56 g, 4.35 mmol, 0.05 eq), Pin2B2 (55.2 g, 217 mmol, 2.5 eq), and potassium acetate (25.6 g, 261 mmol, 3 eq) were added to dioxane (270 mL), and stirred at 80 °C for 6 h under nitrogen protection. The reaction mixture was monitored by LCMS until complete. The reaction solution was poured into water (500 mL) and extracted with ethyl acetate (500 mL * 3). The organic phases were combined, washed with saturated brine (500 mL * 3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain brown solid KH07-4 (33.0 g, crude product). No purification was required; it was directly used in the next reaction step. MS (ESI) m / z: 358.1 [M+H] + .

[0214] Compound KH07-5: Compound KH07-4 (32.0 g, 89.6 mmol, 1 eq) was dissolved in tetrahydrofuran (300 mL), and H2O2 (30.4 g, 268 mmol, 25.8 mL, purity 30.0%, 3 eq) was slowly added dropwise at an external temperature of 0 °C. After the addition was complete, the mixture was stirred at an external temperature of 25 °C for 5 h. The reaction was monitored by LCMS until the starting material was completely reacted. The reaction solution was slowly poured into saturated sodium sulfite (400 mL) to terminate the reaction, and extracted with ethyl acetate (200 mL * 2). The organic phases were combined, washed with saturated brine (100 mL * 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 20 / 1 to 5 / 1) to give white solid KH07-5 (20.0 g, 80.9 mmol, yield 90.3%). MS(ESI) m / z: 248.1 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ 10.6 (s, 1H), 9.63 (s, 1H), 6.87 (d, J = 8.4Hz, 1H), 6.31 (d, J = 8.8Hz, 1H), 4.55 (t, J = 8.8Hz, 2H), 3.09 (t, J = 8.8Hz, 2H).

[0215] Compound KH07-6: Compound KH07-5 (10.0 g, 40.4 mmol, 1 eq) and NCS (6.48 g, 48.5 mmol, 1.20 eq) were added to a mixed solvent of chloroform (100 mL) and DMSO (25.0 mL), and the mixture was stirred at 25 °C for 5 h. The reaction mixture was monitored by LCMS until complete. The reaction solution was directly concentrated and purified by HPLC to give a white solid KH07-6 (4.50 g, 15.9 mmol, yield 39.5%). MS (ESI) m / z: 282.0 [M+H] + . 1 H NMR (CDCl3, 400MHz): δ8.00-7.88 (m, 1H), 7.76 (br s, 1H), 5.57 (br s, 1H), 4.65 (t, J = 8.8Hz, 2H), 3.24-3.18 (m, 2H).

[0216] Compound KH07-7: Compound KH07-6 (2.50 g, 8.88 mmol, 1 eq) and KH07-6a were added to 25 mL of pyridine and reacted at 78 °C with stirring for 5 h. The reaction mixture was monitored by TLC (petroleum ether / ethyl acetate = 5 / 1) to ensure complete reaction. The reaction solution was poured into 60 mL of water and extracted with ethyl acetate (100 mL * 3). The organic phases were combined, washed with saturated brine (100 mL * 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 5 / 1 to 3 / 1) to give a yellow solid KH07-7 (1.20 g, 2.50 mmol, yield 28.1%). MS (ESI) m / z: 482.0 [M+H] + . 1 H NMR(DMSO-d6,400MHz): δ11.20-11.11(m,1H),8.73(s,1H),7.47-7.35(m,1H), 4.68-4.61(m,2H),3.39-3.34(m,1H),3.15-3.06(m,2H),1.15(d,J=6.8Hz,6H).

[0217] Compound KH07-8: Compound KH07-7 (1.05 g, 2.18 mmol, 1 eq), Pin2B2 (1.39 g, 5.46 mmol, 2.5 eq), AcOK (643 mg, 6.55 mmol, 3 eq), and Pd(dppf)Cl2·CH2Cl2 (107 mg, 131 mmol, 0.06 eq) were added to 10 mL of dioxane. The air in the reaction mixture was replaced with nitrogen three times, and the reaction was continued under nitrogen protection at an external temperature of 80 °C with stirring for 12 h. LCMS monitoring showed that there was a residual starting material, and approximately 39.8% of the product was produced. The reaction solution was directly concentrated, and the residue was added to 100 mL of water with stirring. The mixture was extracted with ethyl acetate (100 mL * 2), washed with saturated brine (100 mL * 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a brown solid (1.00 g, crude product). MS(ESI) m / z: 528.1 [M+H] + .

[0218] Compound KH07-9: The synthesis was performed in the same manner as compound KH07-5, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 3 / 1) to give a white solid KH07-9 (700 mg). MS (ESI) m / z: 418.0 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ11.12(s,1H),10.00(s,1H),8.00(s,1H),7.36(s,1H),4.62(t,J=8.8Hz,2H),4 .34(t,J=5.2Hz,1H),3.30-3.26(m,1H),3.02(t,J=8.8Hz,2H),1.17-1.10(m,1H),1.14(d,J=6.8Hz,6H).

[0219] Compound KH07-10: Compound KH07-9 (700 mg, 1.68 mmol, 1 eq) and potassium hydroxide (376 mg, 6.70 mmol, 4 eq) were added to a mixture of 4 mL ethanol and 3 mL water, and the mixture was stirred at 45 °C for 12 h. The reaction mixture was monitored by LCMS to ensure complete reaction. The pH of the reaction solution was adjusted to approximately 7 with 1 N hydrochloric acid, 50 mL of water was added, and the mixture was stirred for 10 min. The solution was then extracted with ethyl acetate (50 mL * 2). The organic phases were combined, washed with saturated brine (100 mL * 2), dried over anhydrous sodium sulfate, filtered, and purified by HPLC to obtain a pale yellow solid KH07-10 (501.4 mg, yield 93.0%, purity 99.6%). MS (ESI) m / z: 322.2 [M+H] + . 1H NMR(DMSO-d6,400MHz): δ9.81(s,1H),7.95(s,1H),6.71-6.35(m,1H),5.75(s,1H),4.78(s,2H),4.50(br t,J=8.8Hz,2H),3.31-3.21(m,1H),2.90(br t,J=8.8Hz,2H),1.12(d,J=6.8Hz,6H).

[0220] Example 8: Synthesis of compound KH07

[0221]

[0222] Compound KH07-11: Reaction Solution A: Weigh compound KH07-10 (202.5 mg, 0.637 mmol) and add it to 10 mL of water. Add 5.6 mL of concentrated hydrochloric acid at 0 °C. Dissolve sodium nitrite (58.3 mg, 0.803 mmol) in 1 mL of water and slowly add it dropwise to the reaction solution. Stir at 0 °C for 1.5 h to form a solution. Reaction Solution B: Weigh compound KH07-10a (108.8 mg, 0.7 mmol) and add it to 20 mL of water. Add 5.6 mL of pyridine at 0 °C and stir at this temperature for 1.5 h. Then, quickly pour reaction solution A into reaction solution B at 0 °C. An orange-red solid is formed. Slowly raise the temperature to room temperature and continue the reaction overnight. TLC monitoring showed that the reaction was complete. Filter the solid directly and wash it with water and petroleum ether (25 mL * 3), respectively. A bright orange-red solid, KH07-11 (280 mg, yield 89.9%, crude product), was obtained. No purification was required; proceed directly to the next step. MS (ESI) m / z: 289.2 [M+H] + .

[0223] Compound KH07: Compound KH07-11 (280 mg, 0.573 mmol) and sodium acetate (485.4 mg, 5.73 mmol) were placed in a single-necked flask, dissolved in acetic acid (10 mL), and protected with N2. The reaction was carried out at 120 °C for 3 h. The reaction was stopped after TLC detection showed complete reaction of the starting material. The reaction was cooled to 0 °C, and after adding 50 mL of water, a large amount of solid precipitated. The solid was directly filtered, and washed with water (20 mL * 3) and petroleum ether (20 mL * 3) respectively. An orange-red solid (254 mg, crude product) was collected, and 50 mg of the crude product was purified by HPLC to obtain a white solid compound KH07 (13.5 mg, 27.0%). MS (ESI) m / z: 443.0 [M+H] + . 1H NMR (DMSO-d6, 400MHz): 10.06 (s, 1H), 7.40 (s, 1H), 4.65 (t, 2H, J = 8.0Hz), 3.21-3.32 (m, 1H), 3.05 (t, 2H, J = 8.0Hz), 1.16 (d, 6H, J = 8.0Hz).

[0224] Example 9: Synthesis of compound KH08

[0225]

[0226] Compound KH08-1: Weigh 200 mg (0.452 mmol) of crude compound KH07 into a single-necked flask, dissolve it in acetic acid (7.5 mL), and then add concentrated hydrochloric acid (2.5 mL) dropwise to the reaction. After reacting at 90 °C for 4 hours, TLC monitoring showed that the starting material had reacted completely and a major polarity point had formed, at which point the reaction was stopped. The reaction solution was directly evaporated to dryness, and the pH of the reaction solution was adjusted to 9-10 with saturated sodium carbonate solution. The solution was extracted twice with ethyl acetate (20 mL * 2), and the aqueous phase was collected. The pH of the aqueous phase was adjusted to 3-4, and the solution was extracted again with ethyl acetate (20 mL * 3). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a yellow solid KH08-1 (186.5 mg, 89.4%), which could be used directly in the next step without further purification.

[0227] Compound KH08: Compound KH08-1 (150 mg, 0.325 mmol, 1 eq) and sodium hydroxide (52 mg, 1.3 mmol, 4 eq) were placed in a single-necked flask, dissolved in water (20 mL), and thioglycolic acid (0.6 g, 6.5 mmol, 20 eq) was added to the reaction solution. The reaction was carried out at 120 °C for 3 h. TLC was used to detect complete reaction of the starting materials. When spots with decreasing polarity were formed, the reaction was stopped. The pH of the reaction system was adjusted to neutral by adding saturated sodium carbonate solution. The mixture was extracted with ethyl acetate (20 mL * 3), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to prepare a white solid KH08 (36.8 mg, 27.1%) after HPLC purification. MS (ESI) m / z: 418.2 [M+H] + .

[0228] Example 10: Synthesis of the key intermediate KH09-6

[0229]

[0230]

[0231] Compound KH09-2: Starting materials KH09-1 (50.0 g, 219 mmol, 28.0 mL, 1.00 eq) and KH09-1a (32.9 g, 219 mmol, 30.8 mL, 1.00 eq) were dissolved in tetrahydrofuran (200 mL). KHMDS (1.00 M, 230 mL, 1.05 eq) was added dropwise at 0 °C. After the addition was complete, the mixture was stirred at this temperature for 10 min, then stirred at 25 °C for 2 h. The reaction was monitored by TLC (petroleum ether / ethyl acetate = 10:1) to ensure complete reaction. The reaction mixture was slowly poured into ice water and extracted with dichloromethane (500 mL * 3). The organic phases were combined, washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, and concentrated to obtain a yellow oily substance KH09-2 (75.0 g, crude product). No purification was required; the product was directly introduced to the next reaction step. 1 H NMR (DMSO-d6, 400MHz): δ8.97(s,1H),7.37-7.28(m,5H),7.26-7.20(m,1H),5.64(s,1H),3.68-3.64(m,3H).

[0232] Compound KH09-3: KH09-2 (75.0 g, 219 mmol, 1.00 eq) was dissolved in 100 mL of acetic acid solution containing hydrogen chloride, and the mixture was stirred at 90 °C for 2 h. The reaction mixture was monitored by LC-MS until complete. The reaction solution was directly evaporated to dryness and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 20 / 1) to give a white solid KH09-3 (23.0 g, 81.1 mmol, yield 36.9%). MS (ESI) m / z: 282.9 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ8.93(s,1H),7.37-7.28(m,2H),7.28-7.21(m,3H),4.23(s,2H).

[0233] Compound KH09-4: Compound KH09-3 (10.0 g, 35.2 mmol, 1.00 eq) and compound KH09-3a (6.28 g, 35.2 mmol, 1.00 eq) were dissolved in 100 mL of DMF, and cesium carbonate (34.4 g, 105 mmol, 3.00 eq) was added. The mixture was stirred at 80 °C for 2 h. The reaction of the starting materials was monitored by TLC (petroleum ether / ethyl acetate = 100:1) to ensure complete reaction. The reaction solution was filtered, and the residue was washed with ethyl acetate (20 mL). The filtrate was collected, 50 mL of water was added, and the mixture was stirred for 5 min. The organic phase was separated, and the aqueous phase was extracted with ethyl acetate (20 mL * 2). The organic phases were combined, washed with saturated brine (100 mL * 2), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 20 / 1) to give a white solid KH09-4 (12.0 g, 28.2 mmol, yield 80.0%). 1 H NMR (DMSO-d6, 400MHz): δ8.75(s,1H),7.41-7.12(m,5H),6.68(s,2H),5.64(s,2H),4.14(s,2H).

[0234] Compound KH09-5: Compound KH09-4 (5.00 g, 11.7 mmol, 1.00 eq), Pin2B2 (5.97 g, 23.5 mmol, 2.00 eq), Pd(dppf)Cl2.CH2Cl2 (480 mg, 588 μmol, 0.05 eq), and potassium acetate (2.31 g, 23.5 mmol, 2.00 eq) were added to 50 mL of dioxane. The air in the reaction solution was replaced with nitrogen three times, and the reaction was stirred at 110 °C for 4 h under nitrogen protection. The reaction mixture was monitored by TLC until the reactants were completely reacted. The reaction solution was filtered, the filtrate was collected, and the solution was directly evaporated to dryness. The solution was purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 100:1 to 50:1) to give a brown oily substance (5.00 g, crude product, containing boric acid products).

[0235] Compound KH09-6: Compound KH09-5 (5.00 g, 10.5 mmol, 1.00 eq) was dissolved in tetrahydrofuran. H2O2 (2.46 g, 21.7 mmol, 2.08 mL, 30% purity, 2.05 eq) was added at 0 °C and stirred for 30 min. Then, H2O2 (4.80 g, 42.3 mmol, 4.07 mL, 30% purity, 4.00 eq) was added and the mixture was stirred at room temperature for 2 h. TLC monitoring showed the reactants had reacted completely. The reaction mixture was poured into 50 mL of saturated sodium sulfite solution and stirred for 15 min. Extraction was performed with dichloromethane (200 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 100:1 to 10:1) to give a pale yellow solid KH09-6 (1.01 g, 2.74 mmol, yield 25.8%). MS (ESI) m / z: 362.0 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ10.03(s,1H),8.01(s,1H),7.38-7.09(m,5H),6.66(s,2H),5.54(s,2H),3.97(s,2H),1.98(s,1H).

[0236] Example 11 Synthesis of compound KH09

[0237]

[0238] Compound KH09-7: The procedure is the same as that for the synthesis of KH07-11, yielding an orange-red solid KH09-7 (323 mg, crude product). No purification is required; proceed directly to the next reaction step.

[0239] Compound KH09: Following the same procedure as compound KH07, an orange-red solid KH09 (432 mg, crude product) was obtained. 50 mg of this crude product was purified by HPLC to yield 6.12 mg of a white solid, with a yield of 7.65%. MS (ESI) m / z: 482.9 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ13.05 -13.10(m,1H),10.26(s,1H),8.07(s,1H),7.75(s,2H),7.18-7.30(m,5H),4.00(s,1H).

[0240] Example 12 Synthesis of compound KH10

[0241]

[0242] Compound KH10-1: The synthesis was performed in the same manner as compound KH08-1, yielding a yellow solid (255.7 mg, crude product). No purification was required; proceed directly to the next reaction. MS (ESI) m / z: 502.0 [M+1] + .

[0243] Compound KH10: Following the same procedure as compound KH08, it yielded a white solid, KH10 (51 mg, yield 20.0%). MS (ESI) m / z: 458.0 [M+1] + . 1 H NMR (DMSO-d6, 400MHz): 12.49 (s, 1H), 10.25 (s, 1H), 8.01 (s, 1H), 7.77 (s, 2H), 7.72 (d, 1H, J = 4.0Hz), 7.17-7.30 (m, 5H), 4.0 (s, 2H).

[0244] Example 13 Synthesis of key intermediate KH11-3

[0245]

[0246] Compound KH11-1: The synthesis was performed in the same manner as compound KH09-4, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 1 / 2) to give an orange-red solid KH11-1 (9.00 g, 26.7 mmol, yield 73.4%). MS (ESI) m / z: 338.1 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ8.42 (s, 1H), 6.43 (s, 2H), 3.52 (br s, 2H), 3.46-3.33 (m, 1H), 2.04 (s, 6H), 1.25 (d, J = 6.8Hz, 6H).

[0247] Compound KH11-2: Compound KH11-1 (5.00 g, 14.8 mmol, 1.00 eq), pin2B2 (7.55 g, 29.7 mmol, 2.00 eq), Pd(dppf)Cl2·CH2Cl2 (607 mg, 743 μmol, 0.05 eq) and potassium acetate (2.92 g, 29.7 mmol, 2.00 eq) were added to 50 mL of dioxane. The air in the reaction liquid was replaced with nitrogen three times, and the reaction was carried out under nitrogen protection at an external temperature of 110 °C for 10 h with stirring. TLC monitoring showed the reactants were completely reacted. The reaction solution was directly evaporated to dryness, and 50 mL of water and 50 mL of ethyl acetate were added. After stirring for 10 min, the organic phase was separated, and the aqueous phase was extracted again with ethyl acetate (50 mL * 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a black oily substance KH11-2 (6.00 g, crude product). No purification was required, and it was directly used in the next reaction step. MS (ESI) m / z: 384.5 [M + H] + .

[0248] Compound KH11-3: Prepared and purified by HPLC (0.1% TFA) as compound KH09-6, yielding an off-white solid (1.11 g, 2.79 mmol, yield 17.8%, TFA salt). MS (ESI) m / z: 274.2 [M+H] + . 1 H NMR (MeOD, 400MHz): δ7.93 (s, 1H), 7.12 (s, 2H), 3.40-3.33 (m, 1H), 2.13 (s, 6H), 1.11 (d, J = 6.8Hz, 6H)

[0249] Example 14 Synthesis of compound KH11

[0250]

[0251] Compound KH11-4: The procedure was the same as for the synthesis of KH07-11, yielding an orange-red solid KH11-4 (368.4 mg, crude product). No purification was required; proceed directly to the next reaction. MS (ESI) m / z: 441.3 [M+H] + .

[0252] Compound KH11: The synthesis was performed in the same manner as compound KH07, yielding an orange-red solid KH11 (342.3 mg, crude product). 50 mg of this crude product was purified by HPLC to give 11.2 mg of a white solid, with a yield of 23.2%. MS (ESI) m / z: 395.2 [M+H] + .

[0253] Example 15 Synthesis of compound KH12

[0254]

[0255] Compound KH12-1: The synthesis was performed in the same manner as compound KH08-1, yielding a yellow solid (286 mg, crude product). No purification was required; proceed directly to the next reaction. MS (ESI) m / z: 414.2 [M+H] + .

[0256] Compound KH10: Following the same procedure as compound KH08, KH10 was given as a white solid (38.4 mg, yield 15.2%). MS (ESI) m / z: 370.1 [M+H] + .

[0257] Example 16 Synthesis of key intermediate KH13-7

[0258]

[0259] Compound KH13-2: N,N-diisopropylamine (1.89 g, 18.6 mmol, 2.63 mL, 0.1 eq) and starting material KH13-1 (25.0 g, 186 mmol, 1 eq) were dissolved in acetonitrile (250 mL), and NCS (26.1 g, 195 mmol, 1.05 eq) was added to the above reaction system. The reaction was stirred at 50 °C for 10 h. The reaction was monitored for completeness by TLC (petroleum ether / ethyl acetate = 10 / 1). The reaction solution was evaporated to dryness and diluted with 100 mL of water, extracted with ethyl acetate (100 mL * 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 20 / 1) to give a yellow solid KH13-2 (5.00 g, 29.6 mmol, yield 15.9%). 1 H NMR (DMSO-d6, 400MHz): δ9.24 (s, 1H), 7.07 (d, J = 8.0 Hz, 1H), 6.69 (d, J = 8.0 Hz, 1H), 2.85-2.77 (m, 4H), 2.05-1.96 (m, 2H).

[0260] Compound KH13-3: Compound KH13-2 (3.70 g, 21.9 mmol, 1 eq) was dissolved in ethanol (37 mL), and nitric acid (2.42 g, 23.0 mmol, 1.73 mL, purity 60.0%, 1.05 eq) was added at 10 °C, and the mixture was stirred at this temperature for 1 h. TLC showed that the starting material had completely disappeared. The reaction solution was diluted with 100 mL of water, extracted with ethyl acetate (50 mL * 3), and the organic phase was collected. The mixture was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 10 / 1) to give a yellow solid KH13-3 (3.5.0 g, 16.3 mmol, yield 74.6%). 1 HNMR(DMSO-d6,400MHz): δ10.96(br s,1H),7.98(s,1H),3.25(t,J=7.6Hz,2H),2.89(t,J=7.6Hz,2H),2.50(td,J=1.6,3.5Hz,1H),2.06(quin,J=7.6Hz,2H).

[0261] Compound KH13-4: Compound KH13-3 (3.50 g, 16.3 mmol, 1 eq) and SnCl2·2H2O (18.4 g, 81.9 mmol, 5 eq) were dissolved in methanol (10 mL) and stirred at 70 °C for 7 h. LCMS analysis showed that the starting material had completely disappeared. The reaction solvent was directly evaporated to dryness, and the residue was dissolved in ethyl acetate. A saturated sodium bicarbonate solution was added, and a solid was produced. The solid was removed by filtration, and the filtrate was collected. The organic phase was separated, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was used directly in the next step without further purification to give a yellow solid KH13-4 (2.70 g, 14.7 mmol, yield 89.7%). 1 H NMR (DMSO-d6, 400MHz): δ 8.12 (s, 1H), 6.39 (s, 1H), 4.48 (s, 2H), 2.75 (t, J = 7.6Hz, 2H), 2.59 (t, J = 7.6Hz, 2H), 2.04-1.89 (m, 2H).

[0262] Compound KH13-5: Compounds KH13-4 (2.50 g, 13.6 mmol, 1 eq), KH07-6a (3.21 g, 13.6 mmol, 1 eq), and cesium carbonate (13.3 g, 40.8 mmol, 3 eq) were added to DMF (30 mL) and stirred at 80 °C for 3 h. TLC (petroleum ether / ethyl acetate = 3 / 1) showed complete disappearance of the starting material. The reaction solution was diluted with 100 mL of water and extracted with ethyl acetate (50 mL * 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 3 / 1) to give a yellow solid KH13-5 (3.70 g, 9.57 mmol, yield 70.3%, purity 99.0%). MS (ESI) m / z: 383.8 [M+H]+. 1 H NMR(DMSO-d6,400MHz): δ8.54-8.35(m,1H),6.70-6.51(m,1H),3.59(s,2H),3.41(spt ,J=6.8Hz,1H),2.76(td,J=7.6,17.5Hz,4H),2.19-2.06(m,2H),1.25(d,J=6.8Hz,6H).

[0263] Compound KH13-6: Following the same procedure as compound KH11-2, a brown solid KH13-6 (4.30 g, crude product) was obtained. MS (ESI) m / z: 430.3 [M+H] + .

[0264] Compound KH13-7: The synthesis was performed in the same manner as compound KH11-3, followed by purification by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 1 / 1) to give a pale yellow solid KH13-7 (1.20 g, 3.72 mmol, yield 38.0%). MS (ESI) m / z: 320.0 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ9.74(s,1H),7.92(s,1H),6.50(s,1H),5.22-4.80(m,2H),3.27(spt,J=6.8Hz,1H),2.64(br t, J=7.6Hz, 2H), 2.59-2.51 (m, 2H), 1.95 (brdd, J=7.6, 15.1Hz, 2H), 1.11 (d, J=6.8Hz, 6H).

[0265] Example 17 Synthesis of compound KH13

[0266]

[0267] Compound KH13-8: The procedure was the same as for the synthesis of KH07-11, yielding an orange-red solid (431.3 mg, crude product). No purification was required; proceed directly to the next reaction. MS (ESI) m / z: 487.2 [M+H] + .

[0268] Compound KH13: Following the same procedure as compound KH07, an orange-red solid KH13 (354.2 mg, crude product) was obtained. 120 mg of this crude product was purified by HPLC to yield 43 mg of a white solid. MS (ESI) m / z: 441.2 [M+H] + . 1 H NMR(DMSO-d6,400MHz):13.07(s,1H),10.06(s,1H),7.98(s,1H),7.38(s,1H),3.25-3.34(m,1 H), 2.84 (t, 2H, J = 8.0Hz), 2.68 (t, 2H, J = 8.0Hz), 2.01 (t, 2H, J = 8.0Hz), 1.15 (d, 6H, J = 4.0Hz).

[0269] Example 18 Synthesis of compound KH14

[0270]

[0271] Compound KH14-1: The synthesis was performed in the same manner as compound KH08-1, yielding a yellow solid (251.4 mg, crude product). No purification was required; proceed directly to the next reaction. MS (ESI) m / z: 460.1 [M+H] + .

[0272] Compound KH14: Following the same procedure as compound KH08, a white solid KH14 (6 mg, yield 11.1%) was obtained. MS (ESI) m / z: 416.2 [M+H] + . 1 H NMR(DMSO-d6,400MHz):12.40(s,1H),9.98(s,1H),7.99(s,1H),7.63(s,1H),7.46(s,1H),3.27-3. 30 (m, 1H), 2.83 (t, 2H, J = 8.0Hz), 2.68 (t, 2H, J = 8.0Hz), 2.0 (t, 2H, J = 8.0Hz), 1.15 (d, 6H, J = 4.0Hz).

[0273] Example 19 Synthesis of key intermediate KH15-8

[0274]

[0275] Compound KH15-2: Sodium hydride (7.49 g, 187 mmol, 60.0% purity, 3.00 eq) was dissolved in tetrahydrofuran (100 mL). KH15-1 (10.0 g, 62.4 mmol, 9.43 mL, 1.00 eq) and CD3I (19.0 g, 131 mmol, 8.16 mL, 2.10 eq) were added to the above reaction system at 0 °C. The reaction was stirred at 0 °C for 0.5 h, then slowly heated to 25 °C and stirred for 11.5 h. The reaction mixture was monitored by TLC to ensure complete reaction of the starting material. The reaction solution was poured into 200 mL of ice water, extracted with ethyl acetate (100 mL * 3), and the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain a yellow oily substance KH15-2 (8.35 g, 42.9 mmol, yield 68.8%). No purification was required; proceed directly to the next reaction.

[0276] Compound KH15-3: Compound KH15-2 (8.95 g, 46.0 mmol, 1.00 eq) was dissolved in water (20 mL), and sodium hydroxide (6.27 g, 111 mmol, 2.42 eq) was added to the above reaction solution. After the addition was complete, the reaction was carried out at 100 °C for 3 h. TLC showed the formation of new spots. The pH was adjusted to 2 with 6N hydrochloric acid at 0 °C, and the mixture was extracted with dichloromethane (100 mL * 2). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The product was used directly in the next step without purification to obtain a colorless oily substance KH15-3 (5.02 g, crude product).

[0277] Compound KH15-4: Compound KH15-3 (5.00 g, 36.1 mmol, 1.00 eq) was heated to 200 °C and stirred for 0.5 h. A new spot was detected by TLC. The crude product was distilled (140 °C, 1 atm) to give a yellow oily substance KH15-4 (2.52 g, 26.7 mmol, yield 73.9%).

[0278] Compound KH15-5: Compound KH15-4 (2.50 g, 28.3 mmol, 1.00 eq), compound KH15-4a (5.49 g, 28.3 mmol, 1.00 eq), silver nitrate (1.64 g, 9.65 mmol, 0.34 eq), and K₂S₂O₈ (11.5 g, 42.5 mmol, 8.52 mL, 1.50 eq) were added to dichloromethane (50 mL) and water (50 mL). The mixture was stirred at 25 °C for 12 h under nitrogen protection. The product spot was detected by TLC (petroleum ether / ethyl acetate = 20 / 1). The reaction solution was quenched with sodium sulfite solution and extracted with dichloromethane (100 mL * 2). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 10 / 1) to obtain the crude product. The crude product was then purified by preparative HPLC to obtain a yellow oily substance KH15-5 (2.60 g, 10.7 mmol, yield 37.9%). MS (ESI) m / z: 243.1 [M+H] + .

[0279] Compound KH15-6: Compound KH15-5 (2.60 g, 10.7 mmol, 1.00 eq), compound KH09-3a (1.92 g, 10.7 mmol, 1.00 eq), and cesium carbonate (10.5 g, 32.29 mmol, 3.00 eq) were added to DMF (26 mL), and the mixture was stirred at 80 °C for 10 h under nitrogen protection. LCMS analysis showed that the starting materials had reacted completely. The reaction mixture was extracted with ethyl acetate (100 mL) and water (100 mL), and the organic phase was collected, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 5 / 1) to give a yellow oily substance KH15-6 (2.70 g, 7.05 mmol, yield 65.4%). MS(ESI) m / z: 384.1 [M+H] + .

[0280] Compound KH15-7: The synthesis was performed in the same manner as compound KH11-2. The residue was purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 5 / 1) to give white solid KH15-7 (3.00 g, 6.97 mmol, yield 98.9%). MS (ESI) m / z = 429.9 [M+H] + .

[0281] Compound KH15-8: Compound KH15-7 (3.00 g, 6.97 mmol, 1.00 eq) was dissolved in tetrahydrofuran (30 mL). Hydrogen peroxide (1.66 g, 14.6 mmol, 1.41 mL, purity 30.0%, 2.10 eq) was added to the reaction mixture at 0 °C. After the addition was complete, the mixture was stirred at room temperature for 8 h. LC-MS analysis showed that the reaction proceeded completely, and the product was detected as MS. The reaction was terminated with saturated sodium sulfite solution (50 mL), and extracted with dichloromethane (50 mL * 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 1 / 1) to give a pale yellow solid KH15-8 (1.20 g, 3.73 mmol, yield 53.5%). MS (ESI) m / z: 320.1 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ9.91-9.78(m,1H),8.01-7.90(m,1H),6.70-6.56(m,2H),5.56-5.42(m,2H),3.25-3.19(m,1H).

[0282] Example 20 Synthesis of compound KH15

[0283]

[0284] Compound KH15-9: The procedure was the same as for the synthesis of KH07-11, yielding an orange-red solid (430 mg, crude product). No purification was required; proceed directly to the next reaction. MS (ESI) m / z: 487.2 [M+H] + .

[0285] Compound KH15: Following the same procedure as compound KH07, 320 mg of crude, orange-red solid KH15 was obtained. 100 mg of this crude solid was purified by HPLC to yield 6 mg of white solid. MS (ESI) m / z: 441.2 [M+H] + .

[0286] Example 21 Synthesis of compound KH16

[0287]

[0288] Compound KH16-1: The synthesis was performed in the same manner as compound KH08-1, yielding a yellow solid (200 mg, crude product). No purification was required; proceed directly to the next reaction. MS (ESI) m / z: 460.1 [M+H] + .

[0289] Compound KH16: Following the same procedure as compound KH08, it yielded a white solid, KH16 (11 mg, yield 21.2%). MS (ESI) m / z: 416.2 [M+H] + .

[0290] Example 22 Synthesis of key intermediate KH17-5

[0291]

[0292] Compound KH17-2: Compound KH17-1 (50.0 g, 238 mmol, 1 eq) and sodium sulfide (27.8 g, 357 mmol, 14.9 mL, 1.5 eq) were added to DMF (500 mL) and stirred at 25 °C for 5 h. LCMS monitoring showed that compound KH17-1 had disappeared. The reaction solution was poured into 500 mL of ice water, and the pH was adjusted to 5 with hydrochloric acid (2N). Extraction was performed with ethyl acetate (200 mL * 2), the organic phases were combined, washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was used directly in the next step without purification to give red solid KH17-2 (43.0 g, crude product). MS (ESI) m / z: 224.1 [M+H] + .

[0293] Compound KH17-3: Compound KH17-2 (40.0 g, 178 mmol, 1 eq) and Zn (58.3 g, 892 mmol, 5 eq) were added to tetrahydrofuran (1.5 L), and acetic acid (21.4 g, 357 mmol, 20.4 mL, 2 eq) was added dropwise at 0 °C. The reaction was stirred at 50 °C for 10 h. LCMS showed complete disappearance of the starting material. The reaction solution was directly filtered, and the filtrate was collected and evaporated to dryness to obtain the residue. The residue was slurried with methyl tert-butyl ether to obtain a white solid KH17-3 (220 g, 108 mmol, yield 60.8%, purity 95.8%). MS (ESI) m / z: 194.0 [M+H] + .

[0294] Compound KH17-4: Compound KH17-3 (16.0 g, 82.4 mmol, 1 eq), compound KH07-6a (16.1 g, 57.7 mmol, 0.7 eq), and cesium carbonate (40.2 g, 123 mmol, 1.5 eq) were added to DMF (200 mL) and stirred at 25 °C for 10 h. LC-MS showed that the starting materials had reacted completely. The reaction solution was added to 200 mL of water and extracted with ethyl acetate (200 mL * 2). The organic phase was collected, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain the residue. The residue was slurried with methyl tert-butyl ether to obtain a yellow solid KH17-4 (2.60 g, 10.7 mmol, yield 37.9%). MS (ESI) m / z: 394.0 [M+H] + .

[0295] Compound KH17-5: Compound KH17-4 (5.00 g, 12.7 mmol, 1 eq) was dissolved in water (25.0 mL) and dioxane (100 mL). Potassium hydroxide (2.85 g, 50.8 mmol, 4 eq), Pd2(dba)3 (1.16 g, 1.27 mmol, 0.1 eq) and t-Buxphos (540 mg, 1.27 mmol, 0.1 eq) were added to the reaction mixture and stirred at 90 °C for 2 h under nitrogen protection. LCMS analysis showed that the starting material had completely disappeared. The pH of the reaction solution was adjusted to 5 with 1M dilute hydrochloric acid, 100 mL of water was added, and extraction was performed with ethyl acetate (100 mL * 2). The organic phase was collected, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 3 / 1) to give an off-white solid KH17-5 (1.20 g, 3.48 mmol, yield 27.3%). MS (ESI) m / z: 330.0 [M + H] + . 1 H NMR (DMSO-d6, 400MHz): δ9.99 (br s, 1H), 8.10-7.90 (m, 1H), 6.83-6.64 (m, 2H), 6.14-5.88 (m, 2H), 3.22 (spt, J = 6.8Hz, 1H), 1.04 (d, J = 6.8Hz, 6H)

[0296] Example 23 Synthesis of compound KH17

[0297]

[0298] Compound KH17-6: The procedure was the same as for the synthesis of KH07-11, yielding an orange-red solid (390 mg, crude product). No purification was required; proceed directly to the next reaction. MS (ESI) m / z: 497.1 [M+H] + .

[0299] Compound KH17: Following the same procedure as compound KH07, 140 mg of crude, orange-red solid KH17 was obtained. 40 mg of this crude product was purified by HPLC to yield 5 mg of a pale yellow solid, with a yield of approximately 12.5%. MS (ESI) m / z: 451.1 [M+H] + .

[0300] Example 24 Synthesis of compound KH18

[0301]

[0302] Compound KH18-1: The synthesis was performed in the same manner as compound KH08-1, yielding a brown solid (102.4 mg, crude product). No purification was required; proceed directly to the next reaction. MS (ESI) m / z: 470.1 [M+H] + .

[0303] Compound KH18: Following the same procedure as compound KH08, a white solid KH18 (18 mg, yield 19.8%) was given. MS (ESI) m / z: 426.0 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): 12.50 (s, 1H), 10.21 (s, 1H), 8.05 (s, 1H), 7.94 (s, 2H), 7.73 (s, 1H), 3.20-3.27 (m, 1H), 1.04 (d, 6H, J = 4.0Hz).

[0304] Example 25 Synthesis of compound KHE001

[0305]

[0306] Compound KHE001-2: Compounds KHE001-1 (20.0 g, 87.8 mmol, 11.2 mL, 1.00 eq) and KHE001-1a (14.8 g, 87.8 mmol, 1.00 eq) were weighed and dissolved in toluene (8 mL). Hexamethyldisilamide potassium (KHMDS) (1 M, 92.2 mL, 1.05 eq) was slowly added dropwise at 0 °C. After stirring the mixture at 20 °C for 2 hours, TLC (petroleum ether / ethyl acetate = 10 / 1) showed that the remaining amount of the starting material was less than 5%, indicating the formation of a principal site with increased polarity. The reaction was quenched in ice water (500 mL) and extracted with ethyl acetate (50 mL * 3). The organic phase was collected, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 30 / 1 to 5 / 1) to give a yellow oil KHE001-2 (17.1 g, yield 54.2%).

[0307] Compound KHE001-3: Compound KHE001-2 (17.0 g, 47.3 mmol, 1.00 eq) was dissolved in hydrochloric acid (20 mL, purity 36%) and acetic acid (80 mL), and stirred at 90 °C for 2 h. TLC (petroleum ether / ethyl acetate = 10 / 1) was used to monitor the reaction until the starting material was completely reacted and a new spot (Rf = 0.60) was formed. The reaction mixture was directly evaporated under reduced pressure to obtain the crude product. The crude product was dissolved in ethyl acetate and washed with saturated sodium bicarbonate aqueous solution (50 mL * 2). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, concentrated to remove the solvent, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1) to give a white solid KHE001-3 (9.70 g, yield 68.0%). 1 H NMR (DMSO-d6, 400MHz): δ8.62 (s, 1H), 7.31 (dd, J = 5.6, 8.8Hz, 2H), 7.04-6.97 (m, 2H), 4.22 (s, 2H).

[0308] Compound KHE001-4: Compounds KHE001-3 (9.00 g, 29.9 mmol, 1.00 eq), KHE001-3a (6.38 g, 35.8 mmol, 1.20 eq), and cesium carbonate (29.2 g, 89.5 mmol, 3.00 eq) were weighed and dissolved in dimethylformamide (90 mL). The reaction system was protected under nitrogen. The mixture was stirred at 80 °C for 2 hours. LCMS monitoring showed that the starting material KHE001-3 was completely consumed, and the target product signal was the main peak. Ethyl acetate (300 mL) and water (100 mL) were added to the reaction solution for extraction and separation. The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to remove the solvent, yielding a crude product. The crude product was purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 30 / 1 to 5 / 1) to give a yellow solid KHE001-4 (4.50 g, yield 34.0%). MS(ESI) m / z: 443.8 [M+H] + .

[0309] Compound KHE001-5: Compound KHE001-4 (4.50 g, 10.1 mmol, 1.0 eq), Pin2B2 (5.16 g, 20.3 mmol, 2.00 eq), Pd(dppf)Cl2·CH2Cl2 (415 mg, 508 μmol, 0.05 eq), and potassium acetate (1.99 g, 20.3 mmol, 2.00 eq) were dissolved in dioxane (45 mL). The reaction system was under nitrogen protection and stirred at 110 °C for 4 hours. LCMS monitoring showed that the starting material KHE001-4 was completely consumed, and the target product signal was the main peak. The reaction system was directly evaporated to dryness to obtain a brown oily substance KHE001-5 (14.0 g, mixture), which was used directly in the next step without purification. MS (ESI) m / z: 490.1 [M+H] + .

[0310] Compound KHE001-6: Compound KHE001-5 (14.0 g, 28.6 mmol, 1.00 eq) was weighed and dissolved in tetrahydrofuran (70 mL) and water (70 mL). Sodium perborate (NaBO3·4H2O) (13.2 g, 85.7 mmol, 3.00 eq) was added to the reaction mixture. The mixture was stirred at 20 °C for 3 hours, and TLC (petroleum ether / ethyl acetate = 2 / 1, Rf = 0.15) showed that the reaction was complete. The reaction mixture was diluted with water (150 mL) and stirred at 25 °C for 10 minutes. The mixture was filtered, and the filter cake was washed with ethyl acetate (20 mL * 2). The combined filtrates were extracted with ethyl acetate (100 mL * 2). The organic phase was collected, washed with saturated brine (300 mL * 2), dried over anhydrous sodium sulfate, and concentrated to remove the solvent, yielding a crude product. The crude product was purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 0 / 1) to give a brown solid KHE001-6 (1.11 g, yield 28.8%, purity 94.2%). MS (ESI) m / z: 380.0 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ10.10-10.05(m,1H),8.02(s,1H),7.30-7.20(m,2H),7.14-7.02(m,2H),6.66(s,2H),5.54(br s,2H),3.99-3.91(m,2H).

[0311] Compound KHE001-7: Reaction Solution A: Compound KHE001-6 (0.5014 g, 1.322 mmol) was weighed and added to 26 mL of water. 14 mL of concentrated hydrochloric acid was added at 0 °C. Sodium nitrite (0.1218 g, 1.765 mmol) was dissolved in 4 mL of water and slowly added dropwise to the reaction solution. The mixture was stirred at 0 °C for 1.5 h to form a solution. Reaction Solution B: Compound KHE001-6a (0.2381 g, 1.526 mmol) was weighed and added to 40 mL of water. 14 mL of pyridine was added at 0 °C, and the mixture was stirred at this temperature for another 1.5 h. Then, reaction solution A was quickly poured into reaction solution B at 0 °C, resulting in the formation of an orange-red solid. The mixture was slowly brought to room temperature and the reaction continued overnight. TLC monitoring showed that the reaction was complete. The solid was directly filtered and washed three times with 50 mL of water and petroleum ether, respectively. A bright orange-red solid, KHE001-7 (600 mg, yield 82.9%, crude product), was obtained. No purification was required; proceed directly to the next step. MS (ESI) m / z: 547.1 [M+H] + .

[0312] Compound KHE001: Compound KHE001-7 (600 mg, 1.097 mmol) and sodium acetate (0.9035 g, 11.018 mmol) were placed in a single-necked flask, dissolved in acetic acid (12 mL), and protected with N2. The reaction was carried out at 120 °C for 3 h. The reaction was stopped after TLC detection showed complete reaction of the starting material. The reaction was cooled to 0 °C, and after adding 100 mL of water, a large amount of solid precipitated. The solid was directly filtered and washed three times with 50 mL of water and petroleum ether, respectively. An orange-red solid (560 mg, crude product) was collected, and 100 mg of the crude product was purified by HPLC to obtain a white solid compound KHE001 (21.8 mg, 23.8%). MS (ESI, m / z): 500.9 [M+1] + . 1 H NMR (DMSO-d6, 400MHz): δ13.25(s,1H),10.28(s,1H),8.08(s,2H),7.74(s,2H),7.22-7.24(m,2H),7.07-7.11(m,2H),3.39(s,2H).

[0313] Example 26 Synthesis of compound KHE002

[0314]

[0315] Compound KHE002-1: Weigh crude compound KHE001 (460 mg, 0.918 mmol) into a single-necked flask, dissolve it in acetic acid (15 mL), and then add concentrated hydrochloric acid (5 mL) dropwise to the reaction. After reacting at 90 °C for 4 hours, TLC monitoring showed that the starting material had reacted completely and a major polarity point had formed, at which point the reaction was stopped. The reaction solution was directly evaporated to dryness, and the pH of the reaction solution was adjusted to 9-10 with saturated sodium carbonate solution. The solution was extracted twice with ethyl acetate (20 mL * 2), and the aqueous phase was collected. The pH of the aqueous phase was adjusted to 3-4, and the solution was extracted again with ethyl acetate (20 mL * 3). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a yellow solid KHE002-1 (300 mg, 62.8%), which could be used directly in the next step without further purification.

[0316] Compound KHE002: Compound KHE002-1 (300 mg, 0.577 mmol) and sodium hydroxide (0.0985 g, 2.462 mmol) were placed in a single-necked flask, dissolved in water (40 mL), and thioglycolic acid (1.0831 g, 11.773 mmol) was added to the reaction solution. The reaction was carried out at 120 °C for 3 h. TLC was used to detect complete reaction of the starting materials. When a point with decreasing polarity was formed, the reaction was stopped. The pH of the reaction system was adjusted to neutral by adding saturated sodium carbonate solution. The mixture was extracted with ethyl acetate (20 mL * 3), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to prepare a white solid KHE002 (46.7 mg, 17.0%) after HPLC purification. MS (ESI, m / z): 476.3 [M+1] + . 1 H NMR (DMSO-d6, 400MHz): δ12.49(s,1H),10.25(s,1H),8.08(s,1H),7.69(s,2H),7.60(s,1H),7.23-7.24(m,2H),7.07-7.12(m,1H),3.98(m,2H).

[0317] Example 27 Synthesis of compound KHE003

[0318]

[0319]

[0320] Compound KHE003-2: KHE003-1a (10.0 g, 42.5 mmol, 1.60 eq) and compound KHE003-1 (5.00 g, 26.6 mmol, 1.00 eq) were dissolved in pyridine (50 mL) and reacted at 130 °C for 5 hours with stirring. TLC (petroleum ether / ethyl acetate = 10 / 1, R f =0.52) Monitor the reaction to ensure it is complete. Cool the reaction to 20°C and dilute with water (500 mL). Extract with ethyl acetate (200 mL * 2). Wash the combined organic phases with saturated brine (300 mL * 2), dry with anhydrous sodium sulfate, filter, concentrate to remove solvent, and obtain crude product. Purify by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 5 / 1) to obtain yellow oil KHE003-2 (2.40 g, yield 23.2%). 1 H NMR (DMSO-d6, 400MHz): δ 8.51-8.47 (m, 1H), 7.73-7.70 (m, 2H), 3.41 (spt, J = 6.8Hz, 1H), 1.19 (d, J = 6.8Hz, 6H).

[0321] Compound KHE003-3: Hydroxylamine hydrochloride (377 mg, 5.43 mmol, 1.50 eq), compound KHE003-2 (1.40 g, 3.62 mmol, 1.00 eq), and sodium bicarbonate (304 mg, 3.62 mmol, 1.00 eq) were added to ethanol (20 mL) and reacted with stirring at 80 °C for 12 hours. TLC (petroleum ether / ethyl acetate = 2 / 1, R f =0.11) The reaction was monitored to ensure completeness. The reaction mixture was directly evaporated to dryness to obtain the crude product, which was then purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 20 / 1) to give a yellow oily substance KHE003-3 (1.00 g, yield 65.8%).

[0322] Compound KHE003-4: Triphosgene (1.61 g, 5.43 mmol, 1.20 eq), compound KHE017-3 (1.90 g, 4.52 mmol, 1.00 eq), and DIEA (2.92 g, 22.6 mmol, 3.94 mL, 5.00 eq) were dissolved in tetrahydrofuran (20 mL) under nitrogen protection and at 0 °C. The reaction mixture was stirred at 0 °C for 0.5 h, then heated to 20 °C and stirred for 12 h. TLC (petroleum ether / ethyl acetate = 1 / 1, R f =0.22) Monitor the reaction to ensure it is complete. Dilute with water (500 mL), and add ethyl acetate (30 mL) and water (150 mL) to the reaction solution for extraction and separation. Collect the organic phase, wash with saturated brine, dry with anhydrous sodium sulfate, concentrate the organic phase to remove the solvent, and obtain the crude product. Purify by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 0 / 1) to give a yellow solid KHE003-4 (1.30 g, yield 64.4%). 1 H NMR (DMSO-d6, 400MHz): δ13.02 (br s, 1H), 8.79 (s, 1H), 8.05 (s, 2H), 3.42-3.40 (m, 1H), 1.13 (d, J = 6.8Hz, 6H).

[0323] Compound KHE003-5: Under nitrogen protection, palladium acetate (60.4 mg, 269 μmol, 0.10 eq), compound KHE003-4 (1.20 g, 2.69 mmol, 1 eq), Pin2B2 (6.83 g, 26.9 mmol, 10.0 eq), and potassium acetate (792 mg, 8.07 mmol, 3.00 eq) were added to DMF (10 mL), and the mixture was stirred at 90 °C for 10 h. The reaction was monitored by LCMS until complete. After cooling the reaction solution to 20 °C, water (100 mL) was slowly added, and the mixture was extracted with ethyl acetate (30 mL * 2). The organic phase was collected, washed with saturated brine (100 mL * 2), dried over anhydrous sodium sulfate, filtered, and the organic solvent was evaporated to obtain a yellow oily substance KHE003-5 (2.00 g, mixture), which was used directly in the next step without purification. MS(ESI)m / z = 493.0 [M+1] + .

[0324] Compound KHE003: Compound KHE003-5 (2.00 g, 4.06 mmol, 1.00 eq) was dissolved in tetrahydrofuran (10 mL) and water (10 mL). Sodium perborate (NaBO3·4H2O) (1.87 g, 12.2 mmol, 3.00 eq) was added to the reaction mixture. The mixture was stirred at 20 °C for 3 hours, and the reaction was monitored by LCMS until complete. The mixture was extracted with ethyl acetate (20 mL) and water (100 mL), and the organic phase was collected. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was evaporated to dryness to obtain the residue. The residue was purified by preparative HPLC to give a yellow solid KHE003 (154 mg, yield 14.9%, purity 97.0%). MS (ESI) m / z: 383.1 [M+1] + . 1 H NMR (DMSO-d6, 400MHz): δ10.12 (br s, 1H), 8.00 (s, 1H), 7.97 (s, 2H), 3.27 (br s, 1H), 1.10 (d, J = 6.8Hz, 6H).

[0325] Example 28 Synthesis of compound KHE004

[0326]

[0327]

[0328] Compound KHE004-2: KHE001-3a (7.56 g, 42.5 mmol, 1.00 eq), compound KHE004-1 (10.0 g, 42.5 mmol, 1.00 eq), and potassium carbonate (11.7 g, 84.9 mmol, 2.00 eq) were added to DMF (100 mL). The mixture was stirred at 80 °C for 3 hours under nitrogen protection. The reaction was monitored for completeness by LC-MS. After cooling the reaction temperature to 20 °C, the mixture was diluted with water (200 mL) and extracted with ethyl acetate (100 mL * 2). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 2 / 1) to give a yellow solid KHE004-2 (12.0 g, yield 75.0%). MS (ESI) m / z = 378.0 [M+1] + .

[0329] Compound KHE004-3: Compound KHE004-2 (5.00 g, 13.3 mmol) and bromoacetonitrile (KHE004-2a) (11.9 g, 99.5 mmol) were dissolved in MeCN (50 mL), and sodium iodide (5.96 g, 39.8 mmol) and potassium carbonate (5.50 g, 39.8 mmol) were added. The mixture was heated to 100 °C in a 100 mL sealed tube and stirred for 48 h. TLC (petroleum ether / ethyl acetate = 5 / 1, R f =0.22) The reaction was monitored to be complete. The reaction was directly evaporated to dryness, and the residue was purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 2 / 1) to give a yellow solid KHE004-3 (4.00 g, yield 72.5%). 1 H NMR(DMSO-d6,400MHz): δ8.72(s,1H),6.92(s,2H),6.77(br t,J=6.8Hz,1H),4.41-4.34(m,1H),4.38(d,J=6.6Hz,2H),3.39-3.34(m,1H),3.39-3.34(m,1H),1.14(d,J=6.8Hz,6H).

[0330] Compound KHE004-4: Compound KHE004-3 (4.00 g, 9.61 mmol) and triethylamine (1.02 g, 10.1 mmol, 1.40 mL) were dissolved in dichloromethane (40 mL), and Boc2O (2.20 g, 10.1 mmol, 2.32 mL) was added. The mixture was stirred at 40 °C for 2 h. TLC (petroleum ether / ethyl acetate = 5 / 1, RL) was performed. f=0.62) to determine if the reaction was complete. The reaction solution was cooled to 20°C and water (200 mL) was slowly added. The mixture was extracted with dichloromethane (40 mL * 2), the organic phase was collected, washed with saturated brine (200 mL * 2), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain a yellow oily substance KHE004-4 (4.80 g, yield 96.7%). 1 H NMR (DMSO-d6, 400MHz): δ8.78 (s, 1H), 7.65 (s, 2H), 4.81 (s, 2H), 3.38-3.33 (m, 1H), 1.46-1.41 (m, 10H), 1.10 (d, J = 6.8Hz, 6H).

[0331] Compound KHE004-5: Compound KHE004-4 (4.80 g, 9.30 mmol) and sodium acetate (6.10 g, 74.4 mmol) were dissolved in DMF (40 mL), and hydroxylamine hydrochloride (5.17 g, 74.4 mmol) was added. The mixture was stirred at 80 °C for 1 h. TLC (petroleum ether / ethyl acetate = 1 / 1, R0) was performed. f =0.39) to detect complete reaction. After cooling to room temperature, extract with ethyl acetate (100 mL) and water (250 mL), collect the organic phase, dry with anhydrous sodium sulfate, filter, evaporate to dryness, and purify by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 0 / 1) to give yellow solid KHE004-5 (4.60 g, yield 90.1%). 1 H NMR (DMSO-d6, 400MHz): δ8.78 (s, 1H), 7.65 (s, 2H), 4.81 (s, 2H), 3.38-3.33 (m, 1H), 1.46-1.41 (m, 9H), 1.10 (d, J = 6.8Hz, 6H).

[0332] Compound KHE004-6: Under nitrogen protection, N,N'-succinimide carbonate (DSC) (2.67 g, 10.4 mmol), compound KHE004-5 (4.40 g, 8.01 mmol), and triethylamine (3.05 g, 30.1 mmol, 4.19 mL) were dissolved in DMF (10 mL) and reacted at 80 °C with stirring for 1 hour. TLC (petroleum ether / ethyl acetate = 2 / 1, R f =0.07) to monitor complete reaction. When the signal of KHE004-6 was detected by LCMS, the reaction solution was separated with ethyl acetate (50 mL) and water (50 mL). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain a yellow solid KHE004-6 (4.20 g, mixture), which was directly used in the next reaction. MS (ESI) m / z: 576.0 [M+1] +.

[0333] Compound KHE004-7: Palladium acetate (39.0 mg, 174 μmol, 0.10 eq), compound KHE004-6 (1.00 g, 1.74 mmol, 1.00 eq), potassium acetate (512 mg, 5.22 mmol, 3.00 eq), and Pin2B2 (4.41 g, 17.4 mmol, 10.0 eq) were dissolved in DMF (15 mL) under nitrogen protection and reacted with stirring at 100 °C for 12 hours. The target product KHE004-7 was detected by LCMS. The reaction solution was extracted with ethyl acetate (20 mL) and water (20 mL), the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain a yellow oily substance KHE004-7 (4.00 g, crude product). This crude product was directly used in the next reaction. MS (ESI) m / z: 622.3 [M+1] + .

[0334] Compound KHE004-8: Compound KHE004-7 (4.00 g, 6.43 mmol, 1.00 eq) was dissolved in tetrahydrofuran (20 mL) and water (20 mL). Sodium perborate (NaBO3·4H2O) (2.97 g, 19.3 mmol, 3.00 eq) was added to the reaction mixture, and the mixture was stirred at 20 °C for 3 hours. The reaction was monitored by LC-MS until complete. The reaction solution was extracted with ethyl acetate (20 mL) and water (100 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain a yellow oily substance KHE004-8 (2.00 g, crude product). This crude product was used directly in the next reaction. MS (ESI) m / z: 514.0 [M+1] + .

[0335] Compound KHE004: Compound KHE004-8 (2.00 g, 3.90 mmol, 1.00 eq) was weighed and dissolved in dichloromethane (20 mL). An ethyl acetate solution of hydrogen chloride (4 M, 10 mL, 10.3 eq) was added to the reaction solution. The mixture was stirred at 20 °C for 3 hours, and the reaction was monitored for completeness by LC-MS. The reaction solution was directly evaporated to dryness and purified by preparative HPLC to obtain a brown solid KHE004 (19.5 mg, yield 1.16%, purity 96.3%). MS (ESI) m / z: 412.0 [M+1] + . 1H NMR(DMSO-d6,400MHz): δ12.42(br s,1H),9.89(br s,1H),7.95(s,1H),6.79(s,1H),6.68-6.55(m,1H),4.29(s,2H),3.32-3.23(m,2H),1.11(s,6H).

[0336] Example 29 Synthesis of compound KHE005

[0337]

[0338] Compound KHE005-1: Palladium acetate (238 mg, 1.06 mmol, 0.10 eq), compound KHE004-2 (4.00 g, 10.6 mmol, 1.00 eq), Pin2B2 (27.0 g, 106 mmol, 10.0 eq), and potassium acetate (3.12 g, 31.8 mmol, 3.00 eq) were dissolved in DMF (100 mL) under nitrogen protection and stirred at 90 °C for 12 hours. The reaction was monitored by LC-MS, and the target peak was clearly visible. The reaction solution was extracted with ethyl acetate (50 mL) and water (50 mL), and the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain a yellow oily substance KHE005-1 (4.00 g, 9.43 mmol, crude product). This crude product was directly used in the next reaction. MS (ESI) m / z: 424.1 [M+H] + .

[0339] Compound KHE005-2: Compound KHE005-1 (4.00 g, 9.43 mmol, 1.00 eq) was dissolved in tetrahydrofuran (10 mL) and water (10 mL). Sodium perborate NaBO3·4H2O (4.35 g, 28.3 mmol, 3.00 eq) was added to the reaction mixture. The mixture was stirred at 20 °C for 3 h. LCMS monitoring showed the reaction was complete, and the target peak was clear. The mixture was extracted with ethyl acetate (20 mL) and water (100 mL), and the organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was evaporated to dryness. The mixture was purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 20 / 1 to 5: / 1) to give a purple oil KHE005-2 (1.40 g, yield 42.0%). MS (ESI) m / z: 314.0 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ10.04 (s, 1H), 7.97-7.99 (m, 3H), 3.27 (d, J = 6.8Hz, 1H), 1.11 (s, 3H), 1.09 (s, 3H).

[0340] Compound KHE005-3: Compound KHE005-2 (800 mg, 2.55 mmol, 1.00 eq) was added to concentrated hydrochloric acid (12.0 M, 20 mL, 94.3 eq), and the reaction was cooled to -5 °C. Sodium nitrite (1.05 g, 15.3 mmol, 6.00 eq) was dissolved in water (4.00 mL) and slowly added dropwise to the above solution while stirring vigorously, maintaining the reaction temperature between -5 and 0 °C. After reacting for 0.5 h, potassium iodide (6.34 g, 38.2 mmol, 15.0 eq) was dissolved in water (6.00 mL) and slowly added dropwise to the above solution while stirring vigorously, continuing to maintain the reaction temperature between -5 and 0 °C. After the addition was complete, the reaction solution was heated to 25 °C and stirred for 12 h under nitrogen protection. The reaction was monitored by LCMS to ensure complete reaction. Extracted and separated by ethyl acetate (20 mL) and water (100 mL), the organic phase was collected, washed with saturated sodium sulfite solution, dried over anhydrous sodium sulfate, filtered, and the organic phase was evaporated to dryness. Purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 20 / 1 to 0 / 1) to give a yellow solid KHE005-3 (546 mg, yield 50.5%). MS (ESI) m / z: 424.9 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ 10.05 (br s, 1H) 7.98 (s, 2H) 7.98 (s, 1H) 3.27 (d, J = 6.8Hz, 1H) 1.11 (s, 3H) 1.10 (s, 3H).

[0341] Compound KHE005-4: Under nitrogen protection, Pd(dppf)Cl2 (93 mg, 0.127 mmol, 0.10 eq), compound KHE005-3 (540 mg, 1.27 mmol, 1.00 eq), Pin2B2 (3.227 g, 12.7 mmol, 10.0 eq), and potassium acetate (374 mg, 3.81 mmol, 3.00 eq) were dissolved in dioxane (7 mL), and the mixture was stirred at 90 °C for 12 h. The reaction was monitored by LCMS until complete. The reaction mixture was directly evaporated to dryness and purified by silica gel column chromatography (SiO2, dichloromethane / methanol = 20 / 1) to obtain the crude product. The crude product was further purified by preparative HPLC to obtain the red solid boric acid product KHE005-4 (400 mg, yield 92.0%). MS (ESI) m / z: 343.0 [M+H] + . 1H NMR (DMSO-d6, 400MHz): δ10.05(s,1H),8.47(s,1H),7.96(s,1H),7.86(s,1H),3.27(d,J=6.8Hz,1H),1.10(s,3H),1.08(s,3H).

[0342] Compound KHE005: Under nitrogen protection, Pd(dppf)Cl2 (51.2 mg, 0.07 mmol, 0.06 eq), compound KHE005-4 (400 mg, 1.17 mmol, 1.00 eq), KHE005-4a (224 mg, 1.17 mmol, 1.00 eq), and potassium phosphate (495 mg, 2.33 mmol, 2.00 eq) were added to a system of dioxane (2.5 mL) and water (2.5 mL), and the mixture was stirred at 100 °C for 12 h. The reaction was monitored by LCMS until complete. The reaction solution was extracted and separated by ethyl acetate (10 mL) and water (80 mL). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was evaporated to dryness. The crude product was purified by silica gel column chromatography (SiO2, dichloromethane / methanol = 20 / 1), and then purified by preparative HPLC to obtain a yellow solid KHE005 (101 mg, yield 21.0%, purity 99.5%). MS (ESI) m / z: 410.1 [M+1] + . 1 H NMR (DMSO-d6, 400MHz): δ12.69 (d, J = 1.2 Hz, 1H), 12.23 (s, 1H), 10.07 (br s, 1H), 8.04-7.97 (m, 3H), 3.36-3.21 (m, 1H), 1.12 (d, J = 6.8 Hz, 6H).

[0343] Example 30 Synthesis of compound KHE006

[0344]

[0345] Compound KHE006-1: Compound KHE005-4a (3.00 g, 15.6 mmol, 1.00 eq) was dissolved in acetonitrile (45 mL). KHE006-1a (8.14 g, 40.0 mmol, 9.89 mL, 2.56 eq) was added to the reaction mixture, and the mixture was stirred at 82 °C for 3 h. Then, iodomethane (2.71 g, 19.1 mmol, 1.19 mL, 1.22 eq) was added to the reaction mixture, and the mixture was stirred for 24 h. Finally, iodomethane (1.11 g, 7.81 mmol, 486 μL, 0.50 eq) was added to the reaction mixture, and the mixture was stirred for another 24 h. TLC (petroleum ether / ethyl acetate = 2 / 1, R0) was performed. f=0.55) indicates that the reaction has been completed. Ethyl acetate (50 mL) and water (50 mL) were added to the reaction solution for extraction. The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 20 / 1 to 0 / 1) to give a yellow solid KHE006-1 (2.00 g, yield 62.1%). 1 H NMR (DMSO-d6, 400MHz): δ3.44 (s, 3H), 12.49 (br s, 1H).

[0346] Compound KHE006: Under nitrogen protection, Pd(dppf)Cl2 (89.6 mg, 0.122 mmol, 0.06 eq), compound KHE005-4 (700 mg, 2.04 mmol, 1.00 eq), KHE006-1 (631 mg, 3.06 mmol, 1.50 eq), and potassium phosphate (866 mg, 4.08 mmol, 2.00 eq) were dissolved in a system of dioxane (2.5 mL) and water (2.5 mL), and the mixture was stirred at 100 °C for 12 h. The reaction was monitored by LCMS to ensure complete reaction. The reaction solution was extracted with ethyl acetate (10 mL) and water (80 mL), the organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was evaporated to dryness. The organic phase was purified by preparative HPLC to obtain a pale yellow solid KHE006 (10.4 mg, yield 1.11%, purity 91.9%). MS(ESI) m / z: 424.1 [M+1] + .

[0347] Example 31 Synthesis of compound KHE007

[0348]

[0349]

[0350] Compound KHE007-2: Compound KHE007-1 (30.0 g, 155 mmol, 1.00 eq) was added to water (300 mL). KHE007-1a (26.5 g, 186 mmol, 26.2 mL, 1.20 eq), silver nitrate (5.27 g, 31.0 mmol, 0.20 eq), and trifluoroacetic acid (8.84 g, 77.6 mmol, 5.74 mL, 0.50 eq) were added to the above reaction system. The reaction mixture was heated to 70 °C and stirred. Ammonium persulfate (NH4)2S2O8 (70.8 g, 310 mmol, 2.00 eq) was slowly added. After the addition was complete, the reaction was continued for 12 h. TLC (petroleum ether / ethyl acetate = 1 / 0, R f=0.12) indicates the reaction is complete. The reaction solution was cooled to 20°C, extracted with ethyl acetate (20 mL) and water (100 mL), and the organic phase was collected. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The solution was purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 10 / 1) to give a colorless oil, KHE007-2 (24.4 g, 84.3 mmol, yield 54.3%). MS (ESI) m / z: 291.2 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ8.88 (s, 1H), 2.74 (d, J = 7.2Hz, 2H), 1.81 (dt, J = 3.6, 7.2Hz, 1H), 1.69-1.54 (m, 5H), 1.27-1.12 (m, 3H), 1.09-0.96 (m, 2H).

[0351] Compound KHE007-3: Compound KHE007-2 (24.4 g, 84.3 mmol, 1.00 eq) and K2CO3 (23.3 g, 169 mmol, 2.00 eq) were added to DMSO (200 mL). Simultaneously, KHE001-3a (15.0 g, 84.3 mmol, 1.00 eq) was added to the above reaction system. The reaction was carried out under nitrogen protection at 60 °C with stirring for 3 h. TLC (petroleum ether / ethyl acetate = 5 / 1, R f =0.34) indicates the reaction was complete. The reaction solution was extracted with ethyl acetate (20 mL) and water (100 mL), the organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was evaporated to dryness. The solution was purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 1 / 1) to give a colorless oil, KHE007-3 (28.0 g, 64.9 mmol, yield 77.1%). MS (ESI) m / z: 432.1 [M+H] + . 1 H NMR(DMSO-d6,400MHz): δ8.71(s,1H),6.68(s,2H),5.62(s,2H),2.68-2.63(m,2 H),1.83-1.69(m,1H),1.66-1.51(m,5H),1.15-1.05(m,3H),1.02-0.88(m,2H).

[0352] Compound KHE007-4: Compound KHE007-3 (2.00 g, 4.64 mmol, 1.00 eq), potassium acetate (911 mg, 9.28 mmol, 2.00 eq), and Pin2B2 (2.36 g, 9.28 mmol, 2.00 eq) were added to dioxane (10 mL). Then, Pd(dppf)Cl2·CH2Cl2 (189 mg, 0.232 mmol, 0.05 eq) was added to the above reaction system. The reaction was carried out under nitrogen protection at 110 °C with stirring for 4 h. The reaction was monitored by LCMS until complete. The reaction solution was extracted with ethyl acetate (20 mL) and water (100 mL). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain a brown solid KHE026-4 (4.00 g, crude product). No purification was required; it was directly used in the next reaction step. MS(ESI) m / z: 478.1 [M+H] + .

[0353] Compound KHE007-5: Crude compound KHE007-4 (4.00 g, 8.36 mmol, 1.00 eq) was dissolved in tetrahydrofuran (20 mL) and water (20 mL). Sodium perborate NaBO3·4H2O (3.86 g, 25.1 mmol, 3.00 eq) was added to the above solution. The reaction was stirred at 20 °C for 3 h. TLC (petroleum ether / ethyl acetate = 2 / 1, R f =0.15) indicates the reaction is complete. The reaction mixture was filtered, the filter cake was washed with ethyl acetate, the filtrate was collected, and extracted with ethyl acetate (50 mL * 2). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was evaporated to dryness. The solution was purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 0 / 1) to give a pale yellow oily product (1.40 g). Further purification by preparative HPLC yielded a gray solid KHE007-5 (1.23 g, 3.27 mmol, yield 72.0%, purity 97.8%). MS (ESI) m / z: 368.1 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ9.74 (s, 1H), 8.00 (s, 1H), 6.65 (s, 2H), 5.51 (s, 2H), 2.94 (s, 1H), 1.62-1.41 (m, 4H), 0.68 (t, J = 7.2Hz, 6H).

[0354] Compound KHE007-6: Reaction Solution A: Compound KHE007-5 (0.5038 g, 1.369 mmol) was weighed and dissolved in 26 mL of water. 14 mL of concentrated hydrochloric acid was added at 0 °C. Sodium nitrite (0.1234 g, 1.788 mmol) was dissolved in 4 mL of water and slowly added dropwise to the reaction solution, maintaining the reaction temperature at 0–5 °C. After the addition was complete, stirring was continued at 0 °C for 1.5 h to form a solution. Reaction Solution B: Compound KHE001-6a (0.2381 g, 1.526 mmol) was weighed and added to 40 mL of water. 14 mL of pyridine was added at 0 °C, and stirring was continued at this temperature for 1.5 h. Then, reaction solution A was quickly poured into reaction solution B at 0 °C, resulting in the formation of an orange-red solid. The temperature was slowly raised to room temperature, and the reaction continued overnight. TLC monitoring showed that the reaction was complete. The solid was directly filtered and washed three times each with 50 mL of water and petroleum ether. The solid was collected to obtain an orange-red solid, KHE007-6 (700 mg, 95.6%, crude product). No purification was required; proceed directly to the next reaction step. MS (ESI) m / z: 535.1 [M+H] + .

[0355] Compound KHE007: Compound KHE007-6 (700 mg, 1.310 mmol) and NaOAc (1.075 g, 13.110 mmol) were placed in a single-necked flask, dissolved in acetic acid (12 mL), and protected under nitrogen. The reaction was carried out at 120 °C for 3 h. The reaction was stopped after TLC detection showed complete reaction of the starting material. The reaction was cooled to 0 °C, and after adding 100 mL of water, a large amount of solid precipitated. The solid was directly filtered and washed three times each with 50 mL of water and petroleum ether. An orange-red solid (620 mg, crude product) was collected. 120 mg of this crude product was sent for preparative HPLC purification to obtain a pale yellow solid compound KHE007 (26.9 mg, yield 22.4%). MS (ESI) m / z: 489.3 [M+1] + . 1 H NMR(DMSO-d6,400MHz): δ13.25(s,1H),10.02(s,1H),8.03(s,2H),7.75(s,1H ),2.49-2.50(m,2H),1.55-1.71(m,6H),1.04-1.13(m,4H),0.92-0.97(m,2H).

[0356] Example 32 Synthesis of compound KHE008

[0357]

[0358] Compound KHE008-1: Weigh compound KHE007 (503.8 mg, 1.030 mmol) into a single-necked flask, dissolve it in acetic acid (15 mL), and then add concentrated hydrochloric acid (5 mL) dropwise to the reaction. After the addition is complete, react at 90 °C for 4 h. TLC monitoring shows that the starting material has reacted completely and a major polarity point has formed, at which point the reaction is stopped. Directly evaporate the reaction solution to dryness, adjust the pH of the reaction to 9-10 with saturated sodium carbonate solution, extract with ethyl acetate (20 mL * 2), collect the aqueous phase, adjust the pH of the aqueous phase to 3-4, extract again with ethyl acetate (20 mL * 2), collect the organic phase, dry with anhydrous sodium sulfate, filter, concentrate the organic phase to remove the solvent to obtain yellow solid KHE008-1 (260 mg, 49.7%), which can be used directly in the next step without purification.

[0359] Compound KHE008: Compound KHE008-1 (260 mg, 0.581 mmol) and sodium hydroxide (0.0891 g, 2.227 mmol) were placed in a single-necked flask, dissolved in water (40 mL), and then thioglycolic acid (1.0125 g, 10.331 mmol) was added to the reaction mixture. The reaction was carried out at 120 °C for 3 h. TLC was used to detect complete reaction of the starting materials, and the formation of spots with decreasing polarity was observed, at which point the reaction was stopped. The pH of the reaction system was adjusted to neutral by adding saturated sodium carbonate solution, extracted with ethyl acetate (20 mL * 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by HPLC to obtain a white solid KHE008 (96.9 mg, 40.9%). MS (ESI) m / z: 464.3 [M+1] + . 1 H NMR(DMSO-d6,400MHz): δ12.46(s,1H),9.99(s,1H),8.03(s,2H),7.77(s,1H),7.69(s,1H),2 .49-2.50(m,2H),1.60-1.71(m,1H),1.56-1.59(m,5H),1.10-1.13(m,3H),0.92-0.97(m,2H).

[0360] Example 33 Synthesis of compound KHE009

[0361]

[0362] Compound KHE009-2: The synthesis was performed in the same manner as compound KHE007-2. The crude product was purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 10 / 1) to give a yellow oil, KHE009-2 (45.5 g, 173 mmol, yield 66.8%). MS (ESI) m / z: 263.0 [M+1] + . 1H NMR (CDCl3, 400MHz): δ8.93 (s, 1H), 3.15-3.05 (m, 1H), 1.73-1.59 (m, 4H), 0.74 (t, J = 7.6Hz, 6H).

[0363] Compound KHE009-3: The synthesis was performed in the same manner as compound KHE007-3. Purification was achieved by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 0 / 1) to give white solid KHE009-3 (11.8 g, 29.1 mmol, yield 38.4%). MS (ESI) m / z: 404.0 [M+1] + . 1 H NMR (DMSO-d6, 400MHz): δ8.77 (s, 1H), 6.68 (s, 2H), 5.61 (s, 2H), 3.04 (t, J = 7.0Hz, 1H), 1.54 (quin, J = 7.2Hz, 4H), 0.69 (t, J = 7.2Hz, 6H).

[0364] Compound KHE009-4: The synthesis was performed in the same manner as compound KHE007-4, yielding a brown solid, KHE009-4 (22.0 g, crude). No purification was required; proceed directly to the next reaction. MS (ESI) m / z: 452.2 [M+H] + .

[0365] Compound KHE009-5: Following the same procedure as compound KHE007-5, a white solid KHE009-5 was obtained (4.44 g, 12.5 mmol, yield 48.7%, purity 96.5%). MS (ESI) m / z: 342.0 [M+H] + .

[0366] 1 H NMR (DMSO-d6, 400MHz): δ9.74 (s, 1H), 8.00 (s, 1H), 6.65 (s, 2H), 5.51 (s, 2H), 2.82-3.01 (m, 1H), 1.62-1.41 (m, 4H), 0.68 (t, J = 7.2Hz, 6H).

[0367] Compound KHE009-6: The synthesis was performed in the same manner as compound KHE007-6, yielding an orange-red solid KHE009-6 (650 mg, 86.7%, crude product). No purification was required; proceed directly to the next reaction step. MS (ESI) m / z: 509.2 [M+H] + .

[0368] Compound KHE009: The synthesis was performed in the same manner as compound KHE007, yielding an orange-red solid (570 mg, crude product). 100 mg of this crude product was purified by preparative HPLC to obtain a pale yellow solid, KHE009 (18 mg, yield 18.2%). MS (ESI) m / z: 463.0 [M+1] + . 1 H NMR(DMSO-d6,400MHz): δ13.26(s,1H),10.00(s,1H),8.06(s,1H),7.75(s, 2H),2.98-3.02(m,1H),1.52-1.55(m,4H),1.24(s,1H),0.68-0.72(m,6H).

[0369] Example 34 Synthesis of compound KHE010

[0370]

[0371] Compound KHE010-1: The synthesis was performed in the same manner as compound KHE008-1, yielding a yellow solid KH010-1 (280 mg, 57.4%), which was used directly in the next step without purification. MS (ESI) m / z: 482.0 [M+1] + .

[0372] Compound KHE010: The synthesis was performed using the same procedure as compound KHE008. After HPLC purification, a white solid, KHE010 (113.6 mg, 44.7%), was obtained. MS (ESI) m / z: 438.2 [M+1] + . 1 H NMR(DMSO-d6,400MHz):12.47(s,1H),9.98(s,1H),8.06(s,1H),7.78(s,2H ), 7.70 (s, 1H), 2.94-3.01 (m, 1H), 1.48-1.61 (m, 4H), 0.70 (t, 6H, J = 8.0Hz).

[0373] Example 35 Synthesis of compound KHE011

[0374]

[0375] Compound KHE011-2: Compounds KHE011-1 (50.0 g, 221 mmol, 1.00 eq), KHE011-1a (30.0 g, 265 mmol, 28.3 mL, 1.20 eq), and Cs2CO3 (144 g, 442 mmol, 2.00 eq) were added to DMF (250 mL) and stirred at 20 °C for 1 h under nitrogen protection.

[0376] TLC (petroleum ether / ethyl acetate = 10 / 1) monitoring showed that the reactants were completely reacted and a new reaction point was formed (R0). f =0.20). The reaction solution was poured into 1N HCl (1L), and a yellow solid precipitated. The mixture was filtered, and the filter cake was washed with water (200mL*3). The filter cake was collected, dissolved in ethyl acetate (1L), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a yellow solid compound KHE011-2 (60.0g, 198mmol, yield 89.7%). No purification was required, and the reaction proceeded directly to the next step. MS (ESI) m / z: 289.0 [M+1] + .

[0377] Compound KHE011-3: Compound KHE001-2 (60.0 g, 198 mmol, 1.00 eq) and LiCl (12.6 g, 297 mmol, 1.50 eq) were added to DMSO (150 mL) and water (50 mL). The mixture was purged with nitrogen three times and reacted at 165 °C for 1 h under nitrogen protection. TLC (petroleum ether / ethyl acetate = 5 / 1) showed complete reaction of the starting material, with a new spot formed (R). f =0.50). The reaction solution was poured into ice water (1 L), extracted with ethyl acetate (500 mL * 3), the organic phase was collected, washed with saturated brine (200 mL * 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 30 / 1 to 5 / 1) to give a yellow solid KHE011-3 (22.1 g, 95.7 mmol, yield 48.3%). MS (ESI) m / z: 231.0 [M+1] + .

[0378] Compound KHE011-4: Compounds KHE011-3 (7.50 g, 32.5 mmol, 1.00 eq), KHE011-3a (9.06 g, 32.5 mmol, 1.00 eq), and Cs₂CO₃ (21.2 g, 64.9 mmol, 2.00 eq) were added to DMSO (70 mL) and stirred at 90 °C for 16 h. TLC (petroleum ether / ethyl acetate = 5 / 1) monitored the reaction of the starting materials until complete, showing slight polarity (R0). f =0.70) A new point was generated. The reaction solution was poured into saturated brine (500 mL), extracted with ethyl acetate (500 mL * 3), the organic phases were combined, washed with saturated brine (200 mL * 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 1 / 1) to give a dark gray solid KHE011-4 (3.95 g, 9.18 mmol, yield 28.3%). MS (ESI) m / z: 429.0 [M+1]+ . 1 H NMR (CDCl3, 400MHz): δ8.66 (s, 1H), 8.29 (s, 2H), 6.39 (s, 1H), 3.43 (d, J = 6.7Hz, 1H), 1.24-1.13 (m, 6H).

[0379] Compound KHE011-5: Compound KHE011-4 (2.90 g, 6.74 mmol, 1.0 eq) was added to a system of water (12 mL), acetic acid (12 mL), and concentrated sulfuric acid (12 mL), and stirred at 110 °C for 6 h. TLC (petroleum ether / ethyl acetate = 5 / 1) was used to monitor the reaction until the reactants were fully reacted and a new spot was observed (R0). f =0.47). The reaction solution was diluted with water (200 mL) and extracted with ethyl acetate (80 mL * 3); the organic phases were combined, washed with saturated brine (200 mL * 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 1 / 1) to give a yellow solid KHE011-5 (2.50 g, 6.17 mmol, yield 91.5%). MS (ESI) m / z: 404.0 [M+1] + . 1 H NMR (DMSO-d6, 400MHz): δ8.79 (s, 1H), 8.35 (s, 2H), 4.62 (s, 2H), 3.38-3.33 (m, 1H), 1.11 (d, J = 6.8Hz, 6H).

[0380] Compound KHE011-6: Following the same procedure as compound KHE007-4, a brown oily substance KHE011-6 (4.50 g, crude) was obtained. No purification was required; it was directly proceeded to the next reaction step. MS (ESI) m / z: 452.2 [M+1] + .

[0381] Compound KHE011-7: The synthesis was performed in the same manner as compound KHE007-5. After HPLC purification, a gray solid KHE011-7 was obtained (680 mg, 1.94 mmol, yield 31.5%, purity 97.8%). MS (ESI) m / z: 342.0 [M+1] + .

[0382] Compound KHE011-8: Compound KHE011-7 (680 mg, 1.95 mmol, purity 97.9%, 1.00 eq) was dissolved in ethanol (10 mL), and stannous chloride (SnCl2·2H2O) (1.32 g, 5.84 mmol, 3.00 eq) was added. The mixture was stirred at 80 °C for 3 h. The reaction mixture was monitored by LCMS until complete. The reaction solution was added in batches to a system of ethyl acetate (30 mL) and water (80 mL). The organic phase was separated, and the aqueous phase was extracted again with ethyl acetate (30 mL * 2). The organic phases were combined, washed with saturated brine (100 mL * 2), dried over anhydrous sodium sulfate, concentrated, and purified by HPLC to obtain a yellow solid KHE011-8 (254 mg, 788 μmol, yield 40.5%, purity 96.8%). MS (ESI) m / z: 312.0 [M+1] + . 1 H NMR (DMSO-d6, 400MHz): δ10.19 (s, 1H), 8.08 (s, 1H), 6.80-6.68 (m, 2H), 4.24 (s, 2H), 3.31-3.23 (m, 1H), 2.08-2.06 (m, 1H), 1.10 (d, J = 6.8Hz, 6H).

[0383] Compound KHE011-9: The synthesis was performed in the same manner as compound KHE007-6, yielding a yellow solid KHE011-9 (350 mg, crude). No purification was required; proceed directly to the next reaction step. MS (ESI) m / z: 479.0 [M+1] + .

[0384] Compound KHE011: Following the same procedure as compound KHE007, after HPLC purification, a yellow solid KHE011 was obtained (194 mg, 433 μmol, yield 63.9%, purity 96.6%). MS (ESI) m / z: 433.0 [M+1] + . 1 H NMR (DMSO-d6, 400MHz): δ13.21 (s, 1H), 10.06 (s, 1H), 8.07 (s, 1H), 7.63 (s, 2H), 4.46 (s, 2H), 3.30-3.26 (m, 1H), 1.11 (d, J = 6.8Hz, 6H).

[0385] Example 36 Synthesis of compound KHE012

[0386]

[0387] Compound KHE012-1: The synthesis was performed in the same manner as compound KHE008-1, yielding a yellow solid KHE012-1 (120 mg, crude product). No purification was required; proceed directly to the next reaction step. MS (ESI) m / z: 451.9 [M+1] + .

[0388] Compound KHE012: The synthesis was performed in the same manner as compound KHE008. After HPLC purification, a pale yellow solid KHE012 (58.0 mg, yield 51.2%, purity 97.9%) was obtained. MS (ESI) m / z: 408.2 [M+1]+. 1 H NMR (DMSO-d6, 400MHz): δ12.43 (s, 1H), 10.05 (m, 1H), 8.07 (s, 1H), 7.69 (d, J = 2 .0Hz,1H),7.66(s,2H),4.44(s,2H),3.35-3.21(m,1H),1.11(d,J=6.8Hz,6H).

[0389] Example 37 Synthesis of compound KHE013

[0390]

[0391] Compound KHE013-2: KHE013-1 (250 g, 1.10 mol, 140 mL, 1.00 eq) and KHE013-1a (165 g, 1.10 mol, 154 mL, 1.00 eq) were dissolved in tetrahydrofuran (2 L). KHMDS (1.0 M, 1.15 L, 1.05 eq) was slowly added dropwise at 0 °C. After the addition was complete, the reaction mixture was heated to 20 °C and stirred for 2 h. TLC (petroleum ether / ethyl acetate = 20 / 1) monitoring showed that the starting material reacted completely, and a new spot (R) was formed. f =0.33). The reaction solution was slowly poured into a saturated ammonium chloride (1L) solution, and extracted with ethyl acetate (600mL*3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a brown solid KHE013-2 (360g, crude product). No purification was required; proceed directly to the next reaction step. MS (ESI) m / z: 343.0 [M+H] + .

[0392] Compound KHE013-3: KHE013-2 (330 g, 966 mmol, 1.00 eq) was added to a solution of concentrated hydrochloric acid (165 mL, 37% purity) and acetic acid (660 mL), and the mixture was stirred at 90 °C for 2 h. TLC (petroleum ether / ethyl acetate = 20 / 1) showed that the starting material reacted completely, and a new spot was formed (R). f=0.48), the solvent was removed by rotary evaporation under reduced pressure, and the remaining oily substance was directly purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 2 / 1) to give white solid KHE013-3 (117 g, 412 mmol, yield 37.6%). 1 H NMR (DMSO-d6, 400MHz): δ8.93 (br d, J=6.0Hz, 1H), 7.37-7.21 (m, 5H), 4.23 (br d, J=2.4Hz, 2H).

[0393] Compound KHE013-4: KHE013-3 (30.0 g, 106 mmol, 1.00 eq) was dissolved in a mixed solution of DMSO (300 mL) and acetic acid (150 mL), and FeCl2·4H2O (2.10 g, 10.6 mmol, 0.10 eq) was added. The mixture was stirred in oxygen at an external temperature of 100 °C for 10 h. TLC (petroleum ether / ethyl acetate = 20 / 1) showed that the starting material reacted completely, and a new spot was formed (R). f =0.24). The reaction solution was slowly poured into ethyl acetate (200 mL) and water (800 mL), the ethyl acetate layer was separated, the aqueous layer was extracted with ethyl acetate (150 mL * 2), the organic layers were combined, washed with saturated brine (500 mL * 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 2 / 1) to give a yellow solid KHE013-4 (16.0 g, 53.8 mmol, yield 50.8%).

[0394] Compound KHE013-5: At an external temperature of -78°C, SF4 (14.2 g, 131 mmol, 3.00 eq) and HF (8.74 g, 437 mmol, 7.95 mL, 10.0 eq) were added to a solution of KHE013-4 (13.0 g, 43.7 mmol, 1.00 eq) in 50 mL of dichloromethane. After the addition was complete, the mixture was stirred at an external temperature of 10°C and 0.30 MPa for 14 h. TLC (petroleum ether / ethyl acetate = 5 / 1) showed that the reaction was complete and a new spot was generated (R). f =0.70). The reaction solution was poured into ethyl acetate (100 mL), and the pH was adjusted to 7-8 with saturated sodium bicarbonate (300 mL). The organic layer was separated, and the aqueous layer was extracted again with ethyl acetate (100 mL * 3). The organic layers were combined, washed with saturated brine (200 mL * 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 1 / 1) to give a colorless oily substance KHE013-5 (10.0 g, 31.3 mmol, yield 71.6%). MS (ESI) m / z: 320.8 [M + H] + .1 H NMR: (CDCl3, 400MHz): δ8.78 (s, 1H), 7.63 (dd, J = 1.2, 7.2Hz, 2H), 7.54-7.41 (m, 3H). 19 F NMR: (CDCl3, 400MHz): δ-96.31.

[0395] Compound KHE013-6: The synthesis was performed in the same manner as compound KHE007-3. Purification was achieved by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 2 / 1) to give a yellow solid KHE013-6 (10.2 g, 22.1 mmol, yield 88.4%). MS (ESI) m / z: 460.0 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ9.03 (s, 1H), 7.64-7.44 (m, 5H), 6.69 (s, 2H), 5.70 (s, 2H).

[0396] Compound KHE013-7: Following the same procedure as compound KHE007-4, a brown oily substance, KHE013-7 (25.0 g, crude), was obtained. No purification was required; proceed directly to the next reaction. MS (ESI) m / z: 508.2 [M+H] + .

[0397] Compound KHE013-8: The synthesis was performed in the same manner as compound KHE007-5. After HPLC purification, KHE013-8 (3.28 g, 8.09 mmol, yield 33.3%, purity 98.2%) was obtained as an off-white solid. MS (ESI) m / z: 380.0 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ10.65(s,1H),8.29(s,1H),7.57-7.43(m,5H),6.67(s,2H). 19 F NMR (DMSO-d6, 400MHz): δ-95.89.

[0398] Compound KHE013-9: The synthesis was performed in the same manner as compound KHE007-6, yielding a yellow solid KHE013-9 (512 mg, crude). No purification was required; proceed directly to the next reaction step. MS (ESI) m / z: 565.0 [M+H] + .

[0399] Compound KHE013: Following the same procedure as compound KHE007, after HPLC purification, KHE013 (134 mg, yield 65.2%, purity 98.1%) was obtained as an off-white solid. MS (ESI) m / z: 519.1 [M+H] + .

[0400] Example 38 Synthesis of compound KHE014

[0401]

[0402] Compound KHE014-1: The synthesis was performed in the same manner as compound KHE008-1, yielding a yellow solid KHE014-1 (63 mg, crude product). No purification was required; proceed directly to the next reaction step. MS (ESI) m / z: 538.0 [M+1] + .

[0403] Compound KHE014: The synthesis was performed using the same procedure as compound KHE008. After HPLC purification, a pale yellow solid, KHE014 (24.0 mg, total yield of 23.2% in both steps, purity 96.8%), was obtained. MS (ESI) m / z: 494.1 [M+1] + .

[0404] Example 39 Synthesis of compound KHE015

[0405]

[0406] Compound KHE015-1: The procedure was the same as that for the synthesis of compound KHE007-3. The compound was purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 1 / 1) to give a yellow solid KHE015-1 (6.00 g, 13.7 mmol, yield 56.5%). 1 H NMR (DMSO-d6, 400MHz): δ9.09 (s, 1H), 7.85-7.75 (m, 3H), 7.62 (t, J = 7.7Hz, 2H), 6.66 (s, 2H), 5.66 (br s, 2H).

[0407] Compound KHE015-2: Compound KHE015-1 (6.00 g, 13.7 mmol, 1.00 eq), potassium hydroxide (3.07 g, 54.7 mmol, 4.00 eq), and t-Bu Xphos (580 mg, 1.37 mmol, 0.10 eq) were added to a system of dioxane (60 mL) and water (15 mL), followed by Pd2(dba)3 (1.25 g, 1.37 mmol, 0.10 eq). After the addition was complete, the solution was placed at 90 °C and stirred for 10 h. TLC (petroleum ether / ethyl acetate = 3 / 1) showed that the reaction was complete and a new spot was generated (R). f =0.22). The reaction solution was poured into ethyl acetate (50 mL) and water (200 mL), stirred for 10 minutes, and the organic phase was separated. The aqueous phase was then extracted with ethyl acetate (50 mL * 2). The organic phases were combined, washed with saturated brine (100 mL * 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by HPLC to obtain a yellow solid KHE015-2 (2.45 g, 6.46 mmol, yield 47.3%, 99.2%). MS (ESI) m / z: 376.0 [M + H] + . 1 H NMR (DMSO-d6, 400MHz): δ10.62(s,1H),8.44(s,1H),7.83-7.77(m,2H),7.72(s,1H),7.56(s,2H),6.64(s,2H),5.84-5.29(m,2H).

[0408] Compound KHE015-3: Following the same procedure as compound KHE007-6, a pale yellow solid KHE015-3 (625.7 mg, crude product) was obtained. No purification was required; proceed directly to the next reaction step. MS (ESI) m / z: 543.0 [M+H] + .

[0409] Compound KHE015: The synthesis of compound KHE007 was performed in the same manner. After HPLC purification, KHE015 (278.4 g, two-step yield 31.1%, 95.9%) was obtained. 1 H NMR (DMSO-d6, 400MHz): 10.87 (s, 1H), 8.49 (s, 1H), 7.82 (s, 1H), 7.80 (d, 1H, J = 4.0Hz), 7.77 (s, 2H), 7.71-7.74 (m, 1H), 7.55-7.59 (m, 2H).

[0410] Example 40 Synthesis of compound KHE016

[0411]

[0412] Compound KHE016-1: The synthesis was performed in the same manner as compound KHE008-1, yielding a yellow solid KHE016-1 (74.5 mg, crude product). No purification was required; proceed directly to the next reaction step. MS (ESI) m / z: 516.0 [M+1] + .

[0413] Compound KHE016: The synthesis was performed in the same manner as compound KHE008. After HPLC purification, KHE016 (31.2 mg, total yield of 26.3% in both steps, purity 97.2%) was obtained as an off-white solid. MS (ESI) m / z: 472.0 [M+1] + .

[0414] Example 41 Synthesis of compound KHE017

[0415]

[0416] Compound KHE017-2: The procedure was the same as for KHE007-2, purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 5 / 1) to give colorless oil KHE017-2 (9.50 g, 40.7 mmol, yield 78.7%). 1 H NMR (CDCl3, 400MHz): δ8.46-8.39 (m, 1H), 2.39 (tt, J = 4.8, 8.0Hz, 1H), 1.25-1.20 (m, 1H), 1.25-1.20 (m, 1H), 1.18-1.11 (m, 2H).

[0417] Compound KHE017-3: Proceeded as with KHE007-3, yielding a gray solid KHE017-3 (13.0 g, crude product). No purification was required; proceed directly to the next reaction. MS (ESI) m / z: 375.9 [M+H] + .

[0418] Compound KHE017-4: Proceeded as with KHE007-4, yielding a yellow solid KHE017-4 (25.0 g, crude product). No purification is required; proceed directly to the next reaction. MS (ESI) m / z: 422.3 [M+H] + .

[0419] Compound KHE017-5: Following the same procedure as KHE007-5, preparative HPLC purification yielded a yellow solid KHE017-5 (439 mg, 1.40 mmol, yield 13.1%, purity 99.4%). MS (ESI) m / z: 312.0 [M+H] + .

[0420] 1 H NMR(DMSO-d6,400MHz): δ9.89(s,1H),7.89(s,1H),6.64(s,1H),6.70-6.60( s,1H),5.52(s,2H),2.39-2.25(m,1H),1.06-0.93(m,2H),0.88-0.76(m,2H).

[0421] Compound KHE017-6: The procedure was the same as for compound KHE007-6, yielding approximately 451.6 mg of an orange-red solid, KHE017-6. No purification was required; proceed directly to the next reaction. MS (ESI) m / z: 479.1 [M+H] + .

[0422] Compound KHE017: Following the same procedure as compound KHE007, 200 mg was used for HPLC purification to obtain approximately 56.2 mg of white solid KHE017. MS (ESI) m / z: 433.0 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ13.27(s,1H),10.12(s,1H),7.94(s,1H),7.74(s,2H),2.28(m,1H),1.04(m,2H),0.86(m,2H).

[0423] Example 42 Synthesis of compound KHE018

[0424]

[0425] Compound KHE018-1: The synthesis was performed in the same manner as compound KHE008-1, yielding a yellowish-brown solid KHE018-1 (205.7 mg, crude product). No purification was required; proceed directly to the next reaction step. MS (ESI) m / z: 452.0 [M+H] + .

[0426] Compound KHE018: The synthesis was performed using the same procedure as compound KHE008. After HPLC purification, KHE018 (20 mg, purity 96.81%) was obtained as an off-white solid. MS (ESI) m / z: 472.0 [M+H] + . 1 H NMR(DMSO-d6,400MHz):12.46(s,1H),10.09(s,1H),8.25(s,1H),7.75(s, 2H),7.68(s,1H),2.32-2.39(m,1H),1.02-1.07(m,2H),0.85-0.90(m,2H).

[0427] Example 43 Synthesis of compound KHE019

[0428]

[0429] Compound KHE019-2: Purified by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 10 / 1) as with KHE007-2, yielding a yellow solid KHE019-2 (9.00 g, 31.7 mmol, yield 68.2%). MS (ESI) m / z: 283.0 [M+H] + . 1 H NMR (CDCl3, 400MHz): δ8.62 (s, 1H), 3.88-3.60 (m, 1H), 3.11-2.90 (m, 4H).

[0430] Compound KHE019-3: Proceeded as with KHE007-3, silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1–10 / 1), yielding a yellow solid KHE019-3 ((5.20 g, 12.2 mmol, yield 86.7%)). MS (ESI) m / z: 424.0 [M+H] + . 1 H NMR (CDCl3, 400MHz): δ8.50 (s, 1H), 6.69 (s, 2H), 4.06-3.75 (m, 2H), 3.75-3.64 (m, 1H), 2.91 (td, J = 8.4, 16.4Hz, 4H).

[0431] Compound KHE019-4: Proceeded as with KHE007-4, yielding a brown oily substance KHE019-4 (5.80 g, crude). No purification was required; proceed directly to the next reaction. MS (ESI) m / z: 472.1 [M+H] + .

[0432] Compound KHE019-5: Following the same procedure as KHE007-5, preparative HPLC purification yielded a white solid KHE019-5 (1.1 g, 2.96 mmol, yield 24.0%, purity 97.4%). MS (ESI) m / z: 362.0 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ10.13 (s, 1H), 8.01 (s, 1H), 6.67 (s, 2H), 5.55 (s, 2H), 3.62 (dquin, J = 2.8, 8.8Hz, 1H), 2.91-2.73 (m, 4H). 19F NMR (DMSO-d6, 400MHz): δ -80.48 (d, J = 12.8Hz, 1F), -95.69 (d, J = 12.8Hz, 1F).

[0433] Compound KHE019-6: Proceeded as with compound KHE007-6, yielding approximately 324 mg of an orange-red solid, KHE019-6. No purification was required; proceed directly to the next reaction. MS (ESI) m / z: 529.0 [M+H] + .

[0434] Compound KHE019: Following the same procedure as compound KHE007, 120 mg was used for HPLC purification to obtain approximately 43.7 mg of white solid KHE019. MS (ESI) m / z: 483.0 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ13.25(s,1H),10.377(s,1H),8.073(s,1H),7.779(s,2H),3.653-3.675(m,1H),2.836-2.948(m,4H).

[0435] Example 44 Synthesis of compound KHE020

[0436]

[0437] Compound KHE020-1: The synthesis was performed in the same manner as compound KHE008-1, yielding a brown solid KHE020-1 (157 mg, crude product). No purification was required; proceed directly to the next reaction step. MS (ESI) m / z: 502.0 [M+H] + .

[0438] Compound KHE020: The synthesis was performed using the same procedure as compound KHE008. After HPLC purification, KHE020 (15.4 mg, purity 97.23%) was obtained as an off-white solid. MS (ESI) m / z: 458.0 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ12.480(s,1H),10.341(s,1H),8.073(s,1H),7.722(s,2H),7.718(s,1H),3.343-3.665(m,1H),2.841-2.946(m,4H).

[0439] Example 45 Synthesis of compound KHE021

[0440]

[0441] Compound KHE021-1: Compound KHE005-2 (0.1095 g, 3.498 mmol, 1.00 eq) was dissolved in THF (10 mL), and KHE021-1a (0.4736 g, 3.882 mmol, 12 eq) and DIEA (0.9965 g, 7.665 mmol, 24 eq) were added to the reaction system. The mixture was stirred overnight at room temperature. New spots were observed during TLC monitoring. The reaction solution was diluted with water, extracted with ethyl acetate (20 mL * 3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain the residue. Purification by TLC plate yielded a yellow solid KHE021-1 (80 mg, yield 57.3%). A sample was sent to LCMS to confirm the product signal. MS (ESI) m / z: 400.0 [M + H] + .

[0442] Compound KHE021: Dissolve compound KHE021-1 (80 mg, 0.2 mmol) in methanol (6 mL), and add sodium hydroxide solution (1 mL, 1 M, aq). Stir overnight at room temperature. The reaction was detected by TLC. Dilute the reaction solution with 10 mL of water, evaporate the organic solvent directly to dryness, adjust the pH to 3–4, extract with ethyl acetate (20 mL * 3), combine the organic phases, dry with anhydrous sodium sulfate, and evaporate to dryness to obtain the residue. Purify the residue by chromatography to give a white solid compound KHE021 (12 mg, yield 15.5%). MS (ESI) m / z: 386.0 [M + H] + . 1 H NMR (DMSO-d6, 400MHz): δ9.87 (s, 1H), 7.94 (s, 1H), 6.65 (s, 2H), 5.53 (s, 2H), 3.23-3.28 (m, 1H), 1.11 (d, J = 6.8Hz, 6H).

[0443] Example 46 Synthesis of compound KHE022

[0444]

[0445]

[0446] Compound KHE022-2: KH01-2 (120 g, 318 mmol, 1.0 eq) was added to hydrochloric acid solution (12 M, 358 mL, 13.5 eq), stirred for 30 minutes, and then cooled to 0°C. Sodium nitrite (24.1 g, 350 mmol, 1.1 eq) was dissolved in 50 mL of water and slowly added dropwise to the above solution. After the addition was complete, the reaction was continued at 0°C with stirring for 1 h. TLC analysis (petroleum ether / ethyl acetate = 5 / 1) showed that the starting material reaction was complete and a new spot was formed (Rf = 0.80). Stannous chloride (215 g, 954 mmol, 3.0 eq) was dissolved in hydrochloric acid (12 M, 493 mL, 18.6 eq) and added dropwise to the above reaction solution at 0°C. After the addition was complete, the reaction was continued at this temperature with stirring for 1 h. LCMS monitoring showed that the starting material reaction was complete and the target product was formed. The reaction solution was filtered, and the filter cake was collected to obtain crude KHE022-2 (80.0 g, 204 mmol, yield: 64.1%), a yellow solid, which required no further purification. MS (ESI) m / z = 392.9 [M+1] + .

[0447] Compound KHE022-3: Compounds KHE022-2 (80.0 g, 204 mmol, 1.0 eq) and KHE022-2A (32.3 g, 367 mmol, 25.8 mL, 1.8 eq) were added to 480 mL of ethanol and 1.50 L of water, and the mixture was stirred at 0 °C for 1 h. LC-MS monitoring showed that KHE022-2 reacted completely, and the target product was formed. The reaction solution was extracted twice with ethyl acetate (2 L * 2), the combined ethyl acetate layers were washed with 500 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a yellow solid KHE022-3 (26.5 g, 57.34 mmol, yield: 28.1%), which did not require purification and proceeded directly to the next reaction. MS (ESI) m / z = 462.9 [M+1] + . 1 H NMR (DMSO-d6, 400MHz): δ1H NMR: DMSO-d6, 400MHz δ 12.16 (s, 1H), 10.04 (s, 1H), 8.75 (s, 1H), 7.56 (s, 2H), 2.06 (s, 3H), 1.24-1.14 (m, 6H).

[0448] Compound KHE022-4: Compound KHE022-3 (10.0 g, 21.64 mmol, 1 eq) was added to 2.0 L of toluene. At room temperature, thionyl chloride (7.72 g, 64.92 mmol, 4.71 mL, 3.0 eq) was added, and the mixture was stirred at 110 °C for 2 h. The reaction mixture was evaporated to dryness to remove excess thionyl chloride, and then dissolved in 2.0 L of toluene. Urea (5.78 g, 64.9 mmol, 3.0 eq) and KHE022-3A (2.89 g, 32.4 mmol, 1.5 eq) were added, and the mixture was stirred at 110 °C for 2 h. LC-MS monitoring showed that the reaction proceeds were completely reacted and the target product was formed. The reaction solution was evaporated to dryness, and 1 L of water and 1 L of ethyl acetate were added. After stirring at room temperature for 30 min, the organic phase was separated. The aqueous phase was extracted once again with 1 L of ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by reversed-phase flash (0.1% TFA) to obtain a yellow solid KHE022-4 (1.00 g, 1.45 mmol, yield: 3.35%, purity: 71.4%). MS (ESI) m / z = 534.0 [M+1] + .

[0449] Compound KHE022-5: KHE022-4 (1.00 g, 1.88 mmol, 1.0 eq) was dissolved in N,N-dimethylacetamide (15 mL), potassium carbonate (777 mg, 5.63 mmol, 3.0 eq) was added, and the mixture was stirred at 120 °C for 6 h. LC-MS monitoring showed that the starting material reacted completely and the target product was formed. 30 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 mL * 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by HPLC to give a yellow solid KHE022-5 (400 mg, 821 μmol, yield: 43.7%). MS (ESI) m / z = 487.9 [M+1] + .

[0450] Compound KHE022-6: KHE022-5 (400 mg, 821 μmol, 1.0 eq) and Pin2B2 (521 mg, 2.05 mmol, 2.5 eq) were dissolved in dioxane (4.0 mL), and potassium acetate (241 mg, 2.46 mmol, 3.0 eq) and Pd(dppf)Cl2·CH2Cl2 (33.5 mg, 41.0 μmol, 0.05 eq) were added. The mixture was stirred at 120 °C for 6 h. LC-MS monitoring showed that the starting material reacted completely. The reaction solution was filtered, and the filtrate was collected. 5 mL of ethyl acetate was added, and the solution was washed successively with 5 mL of water and 5 mL of saturated brine. The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative HPLC to give a yellow solid KHE022-6 (100 mg, 187 μmol, yield: 22.8%). MS(ESI)m / z = 534.1 [M+1] + .

[0451] Compound KHE022: KHE022-6 (90.0 mg, 168 μmol, 1 eq) was added to a mixed solution of tetrahydrofuran (1.0 mL) and water (1.0 mL), followed by sodium perborate (NaBO3·4H2O) (150 mg, 673.91 μmol, 4.0 eq). The mixture was stirred at room temperature for 2 h. LC-MS monitoring showed that the starting material reacted completely and the target product was formed. 5 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (5.0 mL * 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative HPLC to obtain a yellow solid KHE022 (18.0 mg, 42.17 μmol, yield: 25.0%, purity: 99.4%). MS (ESI) m / z = 424.0 [M+1] + . 1 H NMR (DMSO-d6, 400MHz): δ12.40 (s 1H), 10.07 (s, 1H), 8.01 (s, 1H), 7.78 (s, 2H), 3.32-3.26 (m, 1H), 2.17 (s, 3H), 1.16-1.10 (m, 6H).

[0452] Example 47 Synthesis of compound KHE023

[0453]

[0454] Compound KHE023-2: KH01-2 (40.0 g, 106 mmol, 1.00 eq) was added to concentrated sulfuric acid (60.0 mL) and water (25.0 mL). Sodium nitrite solution (7.32 g, 106 mmol, 1.00 eq, dissolved in 150 mL of water) was added dropwise at 0 °C. After the addition was complete, the mixture was stirred at this temperature for 1 h. Copper sulfate solution (253 g, 1.59 mol, 244 mL, 15.0 eq, dissolved in 150 mL of water) was added dropwise to the above reaction solution, along with copper oxide (8.44 g, 106.08 mmol, 1.34 mL, 1.00 eq). The reaction solution was heated to 50 °C and stirred for 12 h. LC-MS monitoring showed the formation of the target product. The reaction solution was filtered, and the filtrate was added to 800 mL of ethyl acetate and washed with 200 mL of saturated saline solution. The organic phase was collected, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by reversed-phase HPLC to give a yellow solid KHE023-2 (7.60 g, 20.0 mmol, yield: 18.96%). MS (ESI) m / z = 378.9 [M+1] + . 1 HNMR (DMSO-d6, 400MHz): δ10.36 (s, 1H), 8.73 (s, 1H), 6.96 (s, 2H), 3.40-3.36 (m, 1H),, 1.12 (d, J = 6.8Hz, 6H).

[0455] Compound KHE023-3: KHE023-2 (4.50 g, 11.9 mmol, 1.00 eq) was dissolved in DMF (50.0 mL), and KHE023-2a (4.60 g, 14.2 mmol, 3.68 mL, 1.20 eq) and cesium carbonate (4.65 g, 14.2 mmol, 1.20 eq) were added. The mixture was stirred at 60 °C for 13 h. LC-MS monitoring showed that the target product was formed. The reactants were poured into 50 mL of water, extracted with ethyl acetate (50 mL * 2), the organic phases were combined, washed with saturated brine (50 mL * 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (petroleum ether / ethyl acetate = 50 / 1 ~ 2 / 1) to give a yellow oil KHE023-3 (3.80 g, 7.19 mmol, yield: 60.4%). MS(ESI)m / z = 529.0 [M+1] + . 1 H NMR (DMSO-d6, 400MHz): δ8.73 (s, 1H), 7.36 (s, 2H), 4.57 (d, J = 9.6Hz, 2H), 4 .18-4.05(m,4H),3.37(m,1H),1.25(t,J=7.2Hz,6H),1.12(d,J=6.8Hz,6H).

[0456] Compound KHE023-4: KHE023-3 (3.50 g, 6.63 mmol, 1.00 eq) was dissolved in dioxane (85.0 mL), and potassium acetate (1.30 g, 13.2 mmol, 2.00 eq), Pin2B2 (3.37 g, 13.2 mmol, 2.00 eq), and Pd(dppf)Cl2 (484 mg, 662 μmol, 0.10 eq) were added. The mixture was stirred at 80 °C for 13 h under nitrogen protection. LC-MS monitoring showed the formation of the target product. The reaction solution was poured into 10 mL of water and extracted with ethyl acetate (10 mL * 2). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a yellow oily crude product, KHE023-4 (2.00 g). No purification was required, and the product was directly introduced to the next reaction step. MS(ESI)m / z = 575.1 [M+1] + .

[0457] Compound KHE023-5: KHE023-4 (2.00 g, 3.48 mmol, 1.00 eq) was dissolved in tetrahydrofuran (40.0 mL), and hydrogen peroxide (3.94 g, 34.7 mmol, 3.34 mL, purity 30.0%, 10.0 eq) was added at 0 °C. The mixture was then stirred at room temperature for 5 h. LC-MS monitoring showed that the starting material reacted completely and the target product was formed. The reaction solution was poured into saturated sodium sulfite (50 mL), stirred for 10 min, and extracted with ethyl acetate (50 mL * 2). The organic phase was collected, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative HPLC to give a brown oily substance KHE023-5 (0.80 g, 1.72 mmol, yield: 49.4%). MS (ESI) m / z = 465.2 [M+1] + . 1 H NMR(DMSO-d6,400MHz): δ10.0(br s,1H),7.98(s,1H),7.32(s,2H),4.57(d,J=9.6Hz,2H),4.13(quin,J=7.2Hz,4H),3.31-3.25(m,1H),1.27(t,J=7.2Hz,6H),1.11(d,J=6.8Hz,6H).

[0458] Compound KHE023: KHE023-5 (0.65 g, 1.40 mmol, 1.00 eq) was dissolved in 20 mL of acetonitrile, and trimethylbromosilane (2.6 g, 16.8 mmol, 2.2 mL, 12.0 eq) was added. The mixture was stirred at room temperature for 3 h. LC-MS monitoring showed the disappearance of the starting material and the formation of the target product. The reactants were poured into 20 mL of methanol, stirred for 30 min, and then evaporated to dryness under reduced pressure. The residue was purified by preparative HPLC to give a white solid KHE023 (0.5 g, 1.22 mmol, yield 87.4%). MS (ESI) m / z = 409.0 [M+H] + . 1 HNMR (DMSO-d6, 400MHz): δ9.96 (br s, 1H), 7.97 (s, 1H), 7.25 (s, 2H), 4.21 (d, J = 10.0Hz, 2H), 3.30 (br d, J = 6.8Hz, 1H), 1.11 (d, J = 6.8Hz, 6H).

[0459] Example 48 In vitro TRα or TRβ binding experiment

[0460] In vitro assays of the agonistic effects of compounds on TRα or TRβ were performed using time-resolved fluorescence resonance energy transfer (FRET) coactivator peptide recruitment assays. These assays employed Europium-anti-GST antibody (Cisbio, 61GSTKLB), biotin-SRC2-2 coactivator peptide (Sangon Biotech), streptavidin-d2 (Cisbio, 610SADAB), RXRα (Pharmaron), and GST-tagged TRα-LBD (Invitrogen, PV4762) or TRβ-LBD (Invitrogen, PV4764). Europium-anti-GST antibody indirectly labeled TRα-LBD or TRβ-LBD by binding to a GST tag. Streptavidin-d2 (Cisbio, 610SADAB) indirectly labeled the SRC2-2 coactivator peptide by binding to a biotin tag. In the presence of RXRα, TRα-LBD or TRβ-LBD can form heterodimers TRα-LBD / RXRα or TRβ-LBD / RXRα, respectively. The agonist binds to TRα-LBD / RXRα or TRβ-LBD / RXRα, leading to a conformational change in TRα-LBD or TRβ-LBD, thereby increasing the recruitment capacity of the heterodimer to the SRC2-2 coactivator peptide. Simultaneously, the resulting decrease in the distance between the d2-labeled SRC2-2 coactivator peptide and the Europium-anti-GST antibody increases the TR-FRET signal. The agonistic activity of compounds can be evaluated based on the effect of different concentrations on TRα or TRβ activity.

[0461] Operating steps:

[0462] a. Prepare 6 mM solutions of the positive compound (MGL-3196) and the test compound (100X) using dimethyl sulfoxide (DMSO), with 100% DMSO as the negative control.

[0463] b. Dilute the positive compound (MGL-3196) or the test compound at a 1:3 ratio from 6 mM (100X) with 100% dimethyl sulfoxide to obtain 10 concentrations, and transfer them into 96-well plates.

[0464] c. Prepare 4X graded dilutions of the compound using 1X reaction buffer (50 mM HEPES (pH 7.0), 50 mM KF, 1 mM DTT, 0.05% NP-40, 0.2% BSA).

[0465] d. Add 5 μl of the 4X compound diluted in a concentration gradient to a 384-well plate.

[0466] e. Prepare 4X TRαLBD and 4X RXRα using 1X reaction buffer (50 mM HEPES (pH 7.0), 50 mM KF, 1 mM DTT, 0.05% NP-40, 0.2% BSA).

[0467] f. Add 5 μl of 4X TRαLBD and 4X RXRα to the 384-well experimental plate.

[0468] g. Prepare a mixture of 2X biotin-SRC2-2, 2X Europium-anti-GST and 2X streptavidin-d2 using 1X reaction buffer (50mM HEPES (pH 7.0), 50mM KF, 1mM DTT, 0.05% NP-40, 0.2% BSA).

[0469] h. Add 10 μl of the 2X mixture (step g) to the 384-well experimental plate.

[0470] i. Incubate at room temperature and in the dark for 4 hours.

[0471] The j.Envision 2104 (PerkinElmer) microplate reader recorded the fluorescence signal values ​​at 665 nm and 615 nm wavelengths in each well of a 384-well experimental plate and calculated the fluorescence ratio at 665 nm / 615 nm.

[0472] Data Analysis:

[0473] Calculate the relative ratio for each well: (ratio 665nm / 615nm - ratio blank), and calculate the percentage of activity (%Activity) as follows:

[0474]

[0475] in:

[0476] Ratio positive This represents the relative comparison of the positive control within the entire plate;

[0477] Ratio vehicle This represents a relative comparison of the negative control within the entire plate.

[0478] Ratio cmpd This represents the relative proportions of compounds throughout the plate.

[0479] The EC was calculated by fitting the relationship between activity (%) and the logarithmic concentration of the compound using Graphpad 8.0 through nonlinear regression. 50 .

[0480] Y = bottom + (top - bottom) / (1 + 10) ^((LogEC 50 -X)*slope))

[0481] X: Logarithmic concentration of compound; Y: Percentage of activity

[0482] Specific test data are shown in Table 1.

[0483] Table 1. Results of in vitro TRα or TRβ binding experiments

[0484]

[0485]

[0486] The positive compound MGL3196 was prepared according to the method described in CN101228135B.

[0487] Example 49 In vitro TRα or TRβ cell transfection experiment

[0488] The aim of this study was to evaluate the agonistic effects of compounds on TRα or TRβ. The coding sequences for TRα-LBD or TRβ-LBD and RXRα-LBD were inserted into the pBIND plasmid (Promega, E1581). Both expression vectors and a reporter vector (pGL4.35 carrying a stably integrated GAL4 promoter-driven luciferase reporter gene) (Promega, E1370) were co-expressed in host cells. When the agonist bound to its corresponding chimeric receptor, the chimeric receptor bound to the GAL4 binding site on the reporter gene vector, stimulating reporter gene expression. The agonistic activity of the compound on TRα or TRβ was determined based on the intensity of the luminescent signal.

[0489] Detailed steps:

[0490] Preparation of working solution

[0491] a) Prepare a 30 mM solution of the reference compound (MGL-3196) or the test compound using dimethyl sulfoxide (DMSO).

[0492] b) All compounds were serially diluted 3-fold with DMSO in 10 concentration gradients, starting at a concentration of 30 mM.

[0493] c) Prepare a positive control of 50 μM T3 (triiodothyronine, prepared by dissolving in DMSO) and a negative control (100% DMSO).

[0494] d) Seal the compound plate and shake for 5 minutes.

[0495] Cell suspension preparation

[0496] a) All cells were cultured according to ATCC standards, and HEK293T cells were used in experiments during the exponential growth phase.

[0497] b) Gently discard the culture medium supernatant. Wash the cells twice with PBS.

[0498] c) Add TrypLE TM Cells were digested with pancreatic enzyme digestion solution (Gibco), and digestion was terminated with complete culture medium (Gibco).

[0499] d) Collect and count the cells; experiments can only be conducted if the cell viability is greater than 90%.

[0500] e) Inoculate with 2.5*10 6 HEK293T cells were placed into each 60mm cell culture dish.

[0501] f) Place the culture dish with inoculated cells in a 37°C, 5% CO2 incubator overnight.

[0502] Cell transfection

[0503] a) Place the Fugene6 transfection reagent (Promega, E2691) at room temperature.

[0504] b) Add 30 μL of Fugene 6 reagent to Opti-MEM TM In the culture medium (Gibco, 11058-021), be careful not to touch the tube wall.

[0505] c) Mix well with a pipette and let stand at room temperature for 5 minutes.

[0506] d) Add 6 μg of plasmid (pBIND plasmid (Pharmaron) with TRα-LBD or TRβ-LBD and RXRα-LBD coding sequences inserted, and pGL4.35 reporter gene plasmid (Promega, E1370)) to the diluted transfection reagent, mix with a pipette, and let stand at room temperature for 20 min.

[0507] e) Add the transfection reagent containing the mixed plasmid DNA to the 60 mm cell culture dish inoculated with the cells.

[0508] f) Place the petri dish in a 37℃, 5% CO2 incubator and incubate for 5 hours.

[0509] Compound treatment

[0510] a) Transfer 50 nmol of each diluted compound solution, positive control, and negative control into a 384-well cell culture plate (PerkinElmer, 6007680-50) using an Echo 550 (Labcyte, 550).

[0511] b) Seed the cells into the 384-well cell culture plate at a density of 15,000 cells per well and add 25 μl of culture medium containing 5% fetal bovine serum (Gibco, 16000-044).

[0512] c) Incubate the cells overnight at 37°C in a 5% CO2 incubator.

[0513] Compound detection

[0514] a) Steady-Glo TM (Promega, E2520) test reagents should be kept at room temperature.

[0515] b) Place the 384-well cell culture plate at room temperature.

[0516] c) Add 25 μL of Steady-Glo to each well. TM The test reagents were applied to the cell culture plate.

[0517] d) Place the culture plate on a shaker and shake it in the dark for 5 minutes.

[0518] The luminescence value was detected using an Envision 2104 (PerkinElmer, Envision HTS).

[0519] Data Analysis:

[0520] The RLU fluorescence signal was calculated for each well, and the percentage of activity (%Activity) was calculated as follows:

[0521] %Activity=(Signal cmpd -Signal Ave_VC ) / (Signal Ave_PC -Signal Ave_VC )×100.

[0522] in:

[0523] Signal ave_pc This represents the average RLU fluorescence signal of the positive controls throughout the entire plate;

[0524] Signal ave_vc This represents the average RLU fluorescence signal of the negative control throughout the entire plate;

[0525] Signa cmpd This represents the average RLU fluorescence signal of the compound throughout the plate.

[0526] The EC was calculated by fitting the relationship between activity (%) and the logarithmic concentration of the compound using Graphpad 8.0 through nonlinear regression. 50 .

[0527] Y = bottom + (top - bottom) / (1 + 10^(LogEC)) 50 -X)*slope))

[0528] X: Logarithmic concentration of compound; Y: Percentage of activity

[0529] The specific test data is shown in Table 2 below.

[0530] Table 2 Results of TRα or TRβ cell transfection experiments

[0531] Compound numbering <![CDATA[TRα(EC 50 μM)]]> <![CDATA[TRβ(EC 50 μM)]]> T3 0.0023 0.002 MGL3196 2.564 1.083 KH03 0.826 0.089 KH04 2.486 1.283 KH06 0.037 0.009 KH07 0.964 0.208 KH10 0.269 0.032 KH13 2.74 1.13 KH14 0.383 0.148 KH15 1.457 0.205 KH16 0.138 0.019 KH18 0.028 0.007 KHE001 0.563 0.258 KHE002 0.039 0.004 KHE007 0.528 0.227 KHE008 0.049 0.008 KHE009 0.507 0.074 KHE011 0.07 0.02 KHE017 3.313 0.88 KHE018 0.407 0.102 KHE021 1.172 0.5

[0532] Example 50 In vitro hepatotoxicity detection

[0533] Primary hepatocyte information:

[0534]

[0535] experiment:

[0536] Step 1: Prepare a 200 mM DMSO stock solution of the test compound, then serially dilute it 3-fold to seven concentrations. Add 1.5 μL of each of the eight concentrations to 498.5 μL of incubation medium (composition shown in Table 5, mix thoroughly) to prepare working solutions. Preheat the incubation medium to 37°C before preparation. DMSO was used as a solvent control. The DMSO content in both the working solution and the solvent control group was 0.3 vol%. The concentrations of each compound are as follows:

[0537]

[0538]

[0539] Step 2: Cell seeding and culture

[0540] 1) Take a tube of donor hepatocytes for cryopreservation, ensuring that the hepatocytes remain frozen before thawing. Quickly place the hepatocytes in a 37°C water bath and gently shake until all ice crystals are dispersed. Spray with 70 vol% ethanol and transfer to a biosafety cabinet.

[0541] 2) Pour the contents of the hepatocyte tubules into a 50 mL centrifuge tube containing 50 mL of resuscitation medium (composition shown in Table 3, mix thoroughly). Centrifuge at 80 g for 8 minutes. After centrifugation, aspirate the resuscitation medium and add inoculation medium (composition shown in Table 4, mix thoroughly). Count the cells using AO / PI staining to obtain a cell density of 0.2 × 10⁶ cells / mL.6 A cell suspension of individual cells.

[0542] 3) Seed the above cell suspension into 100 μL of collagen I-coated 96-well plates. Incubate the plates in an incubator with 5% CO2 and 95% relative humidity for 4-6 hours.

[0543] 4) After incubation for 4-6 hours, observe the cell status under a microscope. Gently shake the culture plate, aspirate the inoculation medium, and add 100 μL of incubation medium to each well (composition shown in Table 5). After incubation for another 18-20 hours, the toxicity test can be performed.

[0544] 5) Before administration, observe cell morphology under a microscope. Aspirate the culture medium from the culture plate and add 100 μL of solvent control (DMSO) or test substance working solution to each well. Test three parallel samples for each condition.

[0545] 6) Every 24 hours after adding the drug, replace the solution with freshly prepared working solution or solvent control solution.

[0546] After 48 hours of treatment with the working solution, the cell morphology was observed under a microscope before use.

[0547] Step 3: Cytotoxicity Detection

[0548] 1) Melt the CellTiter-Glo reagent (supplier Promega, catalog number G9243) stored at -20℃ in a 37℃ water bath.

[0549] 2) After incubating the cell culture plates obtained above for 48 hours, add 50 μL of CellTiter-Glo solution directly to each experimental well.

[0550] 3) Place the cell culture plate in a vortex at 400 rpm for 10 minutes and incubate at room temperature to stabilize the luminescence signal.

[0551] Ten minutes later, 100 μL of reaction solution was aspirated from each well and transferred to a new white non-bottomed translucent plate (Corning 96-well plate, Cat No. 3917). The chemiluminescence value of each well was read using a microplate reader (the chemiluminescence value of the white non-bottomed translucent plate is recorded as "luminescence value"). 空白 The luminescence value of the solvent control is recorded as "luminescence value". 溶媒 ”).

[0552] Step 4: Data Processing

[0553] Cell viability (%) = [(luminescence value)] 待测化合物 -Luminescence value 空白 ) / (luminous value) 溶媒 -Luminescence value 空白)]*100%

[0554] Plot the cell viability (%) versus concentration curve of the compound using GraphPad Prism 8.0.2. Calculate the IC50 of the compound by fitting the cell viability (%) and compound concentration. 50 .

[0555] Y = bottom + (top - bottom) / (1 + 10^(LogIC)) 50 -X)*slope))

[0556] X: Compound concentration; Y: Cell viability (%)

[0557] Specific test data are shown in Table 6.

[0558] Table 3. Resuscitation Culture Medium

[0559]

[0560] Table 4 Inoculation Culture Medium

[0561]

[0562]

[0563] Table 5 Incubation Culture Medium

[0564]

[0565] Table 6. Results of Hepatotoxicity Tests

[0566]

[0567] Example 51: Crab-eating macaque PK experiment

[0568] The purpose of this experiment is to evaluate the pharmacokinetic behavior of the test compound after a single intravenous bolus injection and gavage administration, to examine the bioavailability after gavage administration, and to provide animal experimental data for clinical research.

[0569] Preparation of intravenous bolus and gavage formulations: Accurately weigh an appropriate amount of the test compound and mix it with a suitable solvent (5 vol% DMSO + 10 vol% polyethylene glycol-15-hydroxystearate + 85 vol% physiological saline). After vortexing or sonication, obtain a clear and transparent solution (intravenous administration solution) or a homogeneous suspension. Intravenous bolus formulations should be filtered through a 0.22 μm filter membrane.

[0570] Experimental design: Before the first administration, cynomolgus monkeys were divided into two groups of three males each, based on their body weight. Group 1 animals were administered the test compound (1 mg / kg) via a single intravenous injection (IV); Group 2 animals were administered the test compound (5 mg / kg) via a single oral gavage (PO). The animals were weighed before administration, and the volume of medication administered was calculated based on their body weight.

[0571] Sample Collection: Whole blood samples (approximately 0.2 mL) were collected via upper limb venous puncture at the prescribed time (or other suitable blood collection site), and the actual blood collection time was recorded in the experimental log. The acceptable error for collection time points is ±1 minute for time points within 1 hour of drug administration, and ±5% for other time points. All blood samples were immediately transferred to labeled commercially available centrifuge tubes containing K2-EDTA. After blood sample collection, the samples were centrifuged at 3200g for 10 minutes at 4°C, and the supernatant plasma was aspirated and quickly placed on dry ice and kept at -70±10°C for LC-MS / MS analysis. The collection times for both groups were post-drug administration: 0.083h, 0.25h, 0.5h, 1h, 2h, 4h, 8h, and 24h.

[0572] Data processing: Area under the curve (AUC(0-t) and AUC(0-∞)), elimination half-life (T) 1 / 2 Peak concentration (Cmax), time to reach maximum plasma concentration (Tmax), etc., are calculated using the non-compartmental analysis module in Phoenix WinNonlin 7.0.

[0573] Bioavailability (F) = Area under the curve (AUC) when the drug is administered via PO. (0-t)* dose IV / (Area under curve (AUC) at IV dosing) (0-t) *dose PO )*100%

[0574] Specific data is shown in Table 7.

[0575] Table 7 Results of the Crab-Eating Monkey Comparison Experiment

[0576]

[0577]

[0578] Example 52 In vivo efficacy experiment

[0579] In the early stage of this experiment, DIO (diet-induced obesity) mice were induced by feeding them a high-fat diet. Then, while feeding them a high-fat diet, a NASH model was induced by intraperitoneal injection of CCl4. The anti-NASH efficacy of the test compound was tested in this model.

[0580] Animal information: Male C57BL / 6J mice (approximately 18 weeks old) were used, including 32 DIO mice and 8 normal mice. The DIO mice weighed >38 grams.

[0581] Table 8. Drug Administration Information

[0582]

[0583] Table 9. Grouping Information for Pharmacologically Effective Animals

[0584]

[0585] CCl4 induction: Shake CCl4 thoroughly. Mix 1 part CCl4 with 3 parts olive oil in a glass bottle to prepare a 25% (volume ratio) CCl4 solution. Mix thoroughly and use immediately. The first group of 8 animals served as normal controls and received intraperitoneal injections of physiological saline. Mice in groups 2-5 received intraperitoneal injections of the 25% CCl4 solution twice a week. Administer 25% (volume ratio) CCl4 at a rate of 0.5 ml / kg based on body weight. The injection time of CCl4 should be at least 4 hours apart from the time of administration of other medications on the same day.

[0586] Histopathological analysis

[0587] All liver samples were dehydrated using a Leica HistoCore Pearl-0348 instrument, then embedded using a HistoCoreArcadia embedding machine, and finally sliced ​​using a Leica RM2235 machine.

[0588] NAS rating

[0589] HE-stained (hematoxylin-eosin) sections were scored using the NAS (Neuro-Assay) system, which is the sum of scores for fatty degeneration, ballooning degeneration, and lobular inflammation. Pathologists scored the sections according to the criteria shown in Table 10 below.

[0590] Table 10 NAS Evaluation Methods

[0591]

[0592]

[0593] Lesion assessment criteria:

[0594] (1) Hepatocyte ballooning: Pathological changes resembling vacuoles are observed in hepatocytes. Due to the vacuolar changes, the size of hepatocytes increases, and the hepatocyte nuclei are concentrated or deviated.

[0595] (2) Inflammatory cell infiltration: A large number of inflammatory cells, mainly neutrophils and macrophages, are found in the portal vein area, ventral vein area or around the liver lobules.

[0596] (3) Changes in hepatocyte fat: Regular round vacuoles were observed in hepatocytes of different sizes, with the hepatocyte nucleus located at the edge.

[0597] Percentage of fibrosis

[0598] All Sirius red stained sections were scanned using a Leica Aperio AT2 Brightfield scanner, and the percentage of Sirius red positive staining area was calculated using the HALO AI system to assess the percentage area of ​​Sirius red in the total scanned liver area.

[0599] The results are as follows Figure 1 and Figure 2 As shown.

[0600] Figure 1 The results of the fibrosis evaluation showed that KH06, in a dose-dependent manner, consistently reduced the proportion of fibrosis. KH06 at a dose of 1 mg / kg was more effective than MGL3196 at 3 mg / kg.

[0601] Figure 2 The NAS score results are shown on the vertical axis. The NAS score is the sum of the scores for fatty degeneration, ballooning degeneration, and lobular inflammation. Compared with the model group, KH06 showed a dose-related effect and significantly reduced the scores in all three categories. At the same time, 1 mg / kg of KH06 was as effective as 3 mg / kg of MGL3196.

[0602] For purposes of description and disclosure, all patents, patent applications, and other publications are expressly incorporated herein by reference. These publications are provided solely because their publication predates the filing date of this application. All statements regarding the dates of these documents or representations of their contents are based on information available to the applicant and do not constitute any acknowledgment of the accuracy of the dates or contents of these documents. Furthermore, in any country, any reference to these publications herein does not constitute an endorsement that such publication is part of the general knowledge in the art.

[0603] Those skilled in the art will recognize that the scope of this application is not limited to the various specific implementations and embodiments described above, but rather that various modifications, substitutions, or recombinations can be made without departing from the spirit of this application, all of which fall within the protection scope of this application.

Claims

1. A compound of formula II or a pharmaceutically acceptable salt thereof: Formula II in, R1 is , , , , , , , , , or ; R b and R d For chlorine, R c and R e It is hydrogen; X is -O-; L1 is a single bond; L2 is a single bond; R2 is ; R6 is hydrogen, cyano, COOH, or C. 1-6 alkyl; R7 is hydrogen.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein... R2 is , , or .

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein, R1 is either isopropyl or benzyl.

4. Compounds of the following formula or pharmaceutically acceptable salts thereof: 。 5. A pharmaceutical composition comprising the compound of any one of claims 1-4 or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

6. Use of the compound of any one of claims 1-4 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 5 in the preparation of a medicament for the prevention or treatment of diseases related to the action of β-receptor agonists.

7. The use according to claim 6, wherein the disease associated with the action of the β-receptor agonist is obesity, hyperlipidemia, hypercholesterolemia, thyroid cancer, metabolic syndrome, coronary artery disease, myocardial infarction, ventricular dysfunction, heart failure, cirrhosis, diabetes, atherosclerosis, or hypothyroidism.

8. The use according to claim 6, wherein the disease associated with the β-receptor agonist effect is non-alcoholic steatohepatitis.

9. The use according to claim 6, wherein the disease associated with the action of the β-receptor agonist is dyslipidemia.

10. The use according to claim 6, wherein the disease associated with the action of the β-receptor agonist is hypertriglyceridemia.

11. The use according to claim 6, wherein the disease associated with the action of the β-receptor agonist is a cardiovascular disease.

12. The use according to claim 6, wherein the disease associated with the β-receptor agonist effect is fatty liver.

13. The use according to claim 6, wherein the disease associated with the β-receptor agonist effect is non-alcoholic fatty liver disease.

14. The use according to claim 6, wherein the disease associated with the β-receptor agonist effect is steatohepatitis.

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