Aryl heterocyclic SLC7a11 inhibitor, and preparation method therefor and use thereof

Abstract

An aryl heterocyclic SLC7A11 inhibitor, and a preparation method therefor and the use thereof. The new SLC7A11 inhibitor can be used for preventing and / or treating diseases related to SLC7A11, for example, tumors such as pancreatic cancer, liver cancer, osteosarcoma and glioma, chronic diseases such as fatty liver, cirrhosis, hepatic fibrosis, pulmonary fibrosis and diabetic skin damage, or neurodegenerative diseases such as amyotrophic lateral sclerosis, Parkinson's disease and Alzheimer's disease.

Description

An aromatic heterocyclic SLC7A11 inhibitor and its preparation method and use Technical Field

[0001] The present invention relates to a SLC7A11 inhibitor, a preparation method, and uses thereof. The inhibitor and a pharmaceutical composition containing the same can be used to prevent and / or treat SLC7A11-related diseases or conditions. Background Art

[0002] System xC - It is a sodium-independent antiporter that releases intracellular glutamate and takes up extracellular cystine in a 1:1 ratio (Bannai S (1986) Exchange of cystine and glutamate across plasma membrane of human fibroblasts. J Biol Chem 261:2256-2263). It is involved in glutathione biosynthesis and antioxidant defense. It is overexpressed in a variety of human cancers and is associated with poor prognosis, such as pancreatic cancer, liver cancer, glioma, renal cancer, lung cancer, mesothelioma, and uveal melanoma (Badgley MA, et al. Science. 2020. PMID: 32241947). It consists of two disulfide-linked subunits: the heavy chain subunit solute carrier family 3 member 2 (SLC3A2; also known as CD98 or 4F2hc) and the light chain subunit solute carrier family 7 member 11 (SLC7A11). Studies have shown that tumor cells promote tumor growth by inhibiting ferroptosis by overexpressing SLC7A11. Ferroptosis is a new form of regulated cell death that is iron-dependent and induced by excessive lipid peroxidation.

[0003] In 2020, the number of new cases of pancreatic ductal adenocarcinoma (PDAC) reached 420,000, and the number of deaths reached 410,000. It is estimated that by 2030, pancreatic cancer may become the second leading cause of death among malignant tumors. PDAC is a lethal cancer that is resistant to traditional therapies, with a 5-year survival rate of <9%. Over 90% of PDAC cases carry KRAS mutations, which both promote proliferation and alter cellular metabolism. Mutant KRAS signaling leads to increased production of reactive oxygen species (ROS), which damage cellular components. To compensate, PDAC cells upregulate metabolic programs that use cysteine-derived metabolites, such as glutathione, to detoxify ROS (DeNicola GM et al., Oncogene-induced Nrf2 transcription promotes ROS detoxification and tumorigenesis. Nature 475, 106-109 (2011). [PubMed: 21734707]). Most cellular cysteine ​​is obtained through the system xC-antiporter, which exchanges extracellular oxidized cysteine ​​(cystine) for intracellular glutamate. However, germline deletion of the system xC-gene SLC7A11 is well tolerated in unstressed mice (Sato H et al., Redox imbalance in cystine / glutamate transporter-deficient mice. J. Biol. Chem 280, 37423-37429 (2005). [PubMed: 16144837]), indicating that the basal cystine import requirement of normal cells is low.Studies have shown that SLC7A11 deletion via CRISPR-Cas9 has been shown to induce ferroptosis in cultured PDAC cells and slow the engraftment and growth of xenografts (Daher B et al., Genetic Ablation of the Cystine Transporter xCT in PDAC Cells Inhibits mTORC1, Growth, Survival, and Tumor Formation via Nutrient and Oxidative Stresses. Cancer Res. 79, 3877-3890 (2019). [PubMed: 31175120]), and systemic xC-inhibition has been shown to limit the growth of lymphoma xenografts, inducing lipid oxidation signatures and other indicators of ferroptosis (Zhang Y et al., Imidazole Ketone Erastin Induces Ferroptosis and Slows Tumor Growth in a Mouse Lymphoma Model. Cell Chem. Biol. 26,623-633e629(2019).[PubMed:30799221]).

[0004] Most pancreatic cancer patients are diagnosed at an advanced stage, and currently lack effective therapeutic agents. Most pancreatic cancer treatments, including the standard treatment gemcitabine and novel molecularly targeted therapies, have demonstrated impressive activity against PDAC cells in culture and in some preclinical animal models, but have shown little success in the clinic. This poor prognosis is primarily due to the drug-refractory nature of PDAC, which results from inherent chemoresistance and physical barriers to drug delivery. A dense fibrotic microenvironment drives these mechanisms. Fibrosis alters tumor vasculature, physically hindering drug access and creating a harsh, hypoxic, and nutrient-deprived microenvironment that promotes a more metastatic and chemoresistant mesenchymal phenotype. Pancreatic cancer has a defective vasculature that severely compromises drug delivery. The architects of the pancreatic cancer microenvironment are cancer-associated fibroblasts (CAFs). On the one hand, CAFs are activated by hypoxia signals released by PDAC cells, leading to a self-perpetuating cycle of excessive extracellular matrix (ECM) protein deposition, thereby producing fibrosis. On the other hand, CAFs transmit pro-survival signals to PDAC cells, thereby promoting PDAC cell survival and epithelial-to-mesenchymal transition (DOI: 10.3389 / fphys.2014.00141; DOI: 10.1002 / ags3.12225; DOI: 10.1016 / j.trecan.2019.09.010).

[0005] Based on the aforementioned pathological characteristics of pancreatic cancer, effective drug efficacy requires targeting both pancreatic cancer cells and pancreatic cancer-associated fibroblasts. Studies have demonstrated that high expression of SLC7A11 in the stroma of PDAC tumors, rather than in tumor cells, independently correlates with poor prognosis and overall survival. Knockdown of SLC7A11 in CAFs inhibits their proliferation. Knockdown of SLC7A11 in CAFs significantly inhibits the viability of pancreatic cancer cells and CAFs in co-culture. Knockdown of SLC7A11 in pancreatic cancer cells has no significant effect on tumor growth. However, knockdown of SLC7A11 in both PDAC cells and CAFs significantly inhibits tumor growth, metastasis, and fibrosis. Studies have demonstrated that cancer-associated fibroblasts in pancreatic ductal adenocarcinoma determine the response to SLC7A11 inhibition (Doi:10.1158 / 0008-5472.CAN-20-2496). New drugs are urgently needed to improve patient survival and quality of life. SLC7A11 is also associated with a variety of diseases, including tumors such as liver cancer, osteosarcoma, and glioma; chronic diseases such as fatty liver disease, cirrhosis, liver fibrosis, pulmonary fibrosis, and diabetic skin lesions; and neurodegenerative diseases such as amyotrophic lateral sclerosis, Parkinson's disease, and Alzheimer's disease. Therefore, the development of highly active small molecule SLC7A11 inhibitors has significant social and market value.

[0006] Summary of the Invention

[0007] The inventors are committed to developing a novel SLC7A11 inhibitor, an aromatic heterocyclic compound, which can be used to prevent and / or treat diseases related to SLC7A11, such as pancreatic cancer, liver cancer, osteosarcoma, glioma and other tumors, as well as chronic diseases such as fatty liver, cirrhosis, liver fibrosis, pulmonary fibrosis, diabetic skin lesions, or neurodegenerative diseases such as amyotrophic lateral sclerosis, Parkinson's disease, and Alzheimer's disease.

[0008] The aromatic heterocyclic compound is a compound having a structure represented by formula (I) or a deuterated compound thereof, or a pharmaceutically acceptable salt thereof:

[0009] in,

[0010] X is selected from: N, CR3;

[0011] Optionally, X is: CR3;

[0012] Z1 is independently selected from: C, N;

[0013] Z2 is independently selected from: O, NR6, CR6;

[0014] Dashed lines indicate no chemical bonds or single bonds;

[0015] R1 is selected from the group consisting of: H, alkyl, alkenyl, alkynyl, cycloalkyl, and alicyclic groups optionally substituted with alkyl;

[0016] R2 and R3 are each independently selected from the group consisting of: H, halogen, alkyl, cyano, haloalkyl, alkenyl, alkynyl, cycloalkyl, alicyclic optionally substituted with alkyl, -N(R6)R7, -OR8, -C(O)N(R9)R 10 、 -P(O)(R9)R 10 、 -SO2R';

[0017] R6 and R7 are each independently selected from the group consisting of H, alkyl, haloalkyl, cycloalkyl, alicyclic optionally substituted by alkyl, alkanoyl, haloalkanoyl, cycloalkyl-aminoacyl, -SO2R', or R6 and R7 are linked together with the N group to which they are linked to form alicyclic optionally substituted by alkyl;

[0018] R8 is independently selected from the group consisting of: H, alkyl, haloalkyl, cycloalkyl, alicyclic optionally substituted with alkyl, alkanoyl, haloalkanoyl, heteroaryl;

[0019] R9, R 10 Each is independently selected from: H, alkyl, haloalkyl, cycloalkyl, heterocyclic group optionally substituted by alkyl, or, when R9 and R 10 When connected to the same N or -P(O), R9 and R 10 It can also form an aliphatic heterocyclic group optionally substituted by an alkyl group together with the N or -P(O) to which it is commonly connected;

[0020] Each R' is independently selected from the group consisting of: alkyl, haloalkyl, cycloalkyl, alicyclic, amino, alkylamino, and cycloalkyl-amino;

[0021] R4 is selected from the group consisting of optionally substituted alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, and alicyclic groups, wherein the substituent in the optionally substituted alkyl is selected from the group consisting of halogen, cyano, hydroxy, and alkoxy;

[0022] R5 is selected from -CF3, acetyl;

[0023] Optionally, R5 is selected from Alternatively, R2 and R5 together with the benzene ring to which they are attached form the following structure:

[0024] Each Ra is independently selected from alkyl, haloalkyl;

[0025] Optionally, R5 is selected from

[0026] The substituents in the "optionally alkyl-substituted alicyclic group", "haloalkanoyl", "haloalkyl" and "optionally substituted alkyl" are 1 or more, preferably 1, 2 or 3;

[0027] Optionally, the compound represented by formula (I) is in the form of its tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture of these isomers.

[0028] Optionally, the halogen atoms in the "halogen", "haloalkyl" and "haloalkanoyl" are independently selected from F, Cl, Br and I;

[0029] Alternatively, the alkyl groups in the “alkyl”, “haloalkyl”, “alkoxy”, “alkanoyl”, “haloalkanoyl” and “heterocyclic group optionally substituted by an alkyl group” are each independently C1-C 10 linear or branched alkyl; optionally, C1-C7 linear or branched alkyl; optionally, C1-C5 linear or branched alkyl; optionally, selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, isopentyl, 1-ethylpropyl, neopentyl, n-hexyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl ... hexyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 3-ethylpentyl, or 2,2,3-trimethylbutyl;

[0030] Optionally, the "alkenyl" is a C2-C 15 Straight or branched alkenyl, optionally C2-C 10 Straight chain or branched alkenyl, optionally C2-C8 straight chain or branched alkenyl, optionally C2-C6 straight chain or branched alkenyl, optionally C2-C4 straight chain or branched alkenyl, optionally selected from ethenyl, propenyl, butenyl, pentenyl;

[0031] Optionally, the "alkynyl" group contains one or more triple bonds. 15 Straight or branched chain alkynyl, optionally C2-C 10 Straight chain or branched chain alkynyl, optionally, C2-C8 straight chain or branched chain alkynyl, optionally, C2-C6 straight chain or branched chain alkynyl, optionally, C2-C4 straight chain or branched chain alkynyl; optionally, selected from ethynyl, propynyl, hexynyl;

[0032] Optionally, the cycloalkyl group in the "cycloalkyl group", "cycloalkyl-aminoacyl group" and "cycloalkyl-amino group" is C3-C 10 Monocyclic or bicyclic cycloalkyl, optionally, is a C3-C7 monocyclic cycloalkyl, optionally, selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl;

[0033] Alternatively, the alicyclic group in the “alicyclic group”, “alicyclic group optionally substituted by an alkyl group” and “alicyclic-amino group” is a 3-10 membered non-aromatic heterocycle containing 1, 2 or 3 heteroatoms selected from N, O and S on the ring; Alternatively, the alicyclic group is a 3-10 membered non-aromatic heterocycle containing 1 or 2 heteroatoms selected from N and O on the ring; Alternatively, the alicyclic group is a 3-6 membered non-aromatic heterocycle containing 1 or 2 heteroatoms selected from N and O on the ring; Alternatively, the alicyclic group is a 3-10 membered non-aromatic heterocycle containing 1 or 2 heteroatoms selected from N and S on the ring; Alternatively, the alicyclic group is a 3-10 membered non-aromatic heterocycle containing 1 or 2 heteroatoms selected from N and S on the ring; The heterocycle is a 3-6 membered non-aromatic heterocycle containing 1 or 2 heteroatoms selected from N and S on the ring; optionally, the heterocyclic group in the "heterocyclic group", "heterocyclic group optionally substituted by alkyl" and "heterocyclic group-amino" is selected from methyl-substituted azetidinyl, methyl-substituted azetidinyl, methyl-substituted azetidinyl, methyl-substituted azetidinyl, azetidinyl, azetidinyl, oxolanyl, oxolanyl, azetidinyl, piperidinyl, piperazinyl and alkenylpiperidine; optionally, the heterocyclic group in the "heterocyclic group", "heterocyclic group optionally substituted by alkyl" and "heterocyclic group-amino" is selected from methyl-substituted azetidinyl, ...

[0034] Optionally, the heteroaromatic ring of the "heteroaryl" is a 5-10 membered heteroaromatic ring containing 1-2 heteroatoms selected from N and O on the ring; optionally, the heteroaryl is selected from pyridyl, pyrrolyl, pyrazolyl, thiazolyl, oxazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, naphthyl, quinolyl, quinazolinyl; optionally, the heteroaryl is selected from pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, pyrimidin-6-yl.

[0035] Optionally, the compound represented by the above formula (I) or its deuterated form, or a pharmaceutically acceptable salt thereof, is selected from the following compounds:

[0036] On the other hand, a method for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided, characterized in that the method comprises the following reaction scheme:

[0037] Compound I-1 was prepared using the following method I:

[0038] In each formula, R1, R2, R3, R4, and R5 are as defined in formula (I).

[0039] The synthesis reaction of Ib can be carried out in the presence of a reaction reagent, an activating agent, and a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the activating agent can be selected from, but not limited to, phosphorus trichloride, phosphorus oxychloride, phosphorus pentachloride, or the like; and the base can be selected from, but not limited to, N,N-diisopropylethylamine, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, pyridine, or the like.

[0040] The synthesis reaction of Id can be carried out in the presence of a reaction reagent, a catalyst, and a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the catalyst can be selected from, but not limited to, sodium iodide, potassium iodide, etc.; the base can be selected from, but not limited to, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, N,N-diisopropylethylamine, pyridine, etc.

[0041] The synthesis reaction of I-1 can be carried out in the presence of a reaction reagent and a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the base can be selected from, but not limited to, N,N-diisopropylethylamine, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, pyridine, and the like.

[0042] Alternatively, compound I-1 can be synthesized using the following method II:

[0043] In each formula, R1, R2, R3, R4, and R5 are as defined in formula (I).

[0044] The synthesis reaction of Ib can be carried out in the presence of a reaction reagent, an activating agent, and a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the activating agent can be selected from, but not limited to, phosphorus trichloride, phosphorus oxychloride, phosphorus pentachloride, or the like; and the base can be selected from, but not limited to, N,N-diisopropylethylamine, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, pyridine, or the like.

[0045] The synthesis reaction of II-a can be carried out in the presence of a reaction reagent, a catalyst, and a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the catalyst can be selected from, but not limited to, sodium iodide and potassium iodide; and the base can be selected from, but not limited to, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, N,N-diisopropylethylamine, pyridine, and the like.

[0046] The synthesis reaction of II-b can be carried out in the presence of a reaction reagent, an acid, or a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the acid can be selected from, but not limited to, hydrochloric acid, trifluoroacetic acid, acetic acid, boric acid, sulfuric acid, and the like; and the base can be selected from, but not limited to, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, N,N-diisopropylethylamine, pyridine, and the like.

[0047] The synthesis reaction of I-1 can be carried out in the presence of a reaction reagent and a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the base can be selected from, but not limited to, N,N-diisopropylethylamine, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, pyridine, and the like.

[0048] Alternatively, compound III-1 may be synthesized using the following method III:

[0049] In each formula, R1, R2, R3, R4, and R5 are as defined in formula (I).

[0050] The synthesis reaction of III-c can be carried out in the presence of a reaction reagent, a condensation reagent, an activation reagent, and a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the condensation reagent can be selected from, but not limited to, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, carbonyldiimidazole, etc.; the activation reagent can be selected from, but not limited to, 1-hydroxy-7-azabenzotriazole, etc.; the base can be selected from, but not limited to, N,N-diisopropylethylamine, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, pyridine, etc.

[0051] The synthesis reaction of III-d can be carried out in the presence of a reaction reagent, a catalyst, and a base in a solvent for coupling reaction. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, ethanol, DME, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the catalyst can be selected from, but not limited to, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex, [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium, tetrakis(triphenylphosphine)palladium, methanesulfonic acid (2-dicyclohexylphosphine) The alkali metals include but are not limited to potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, pyridine, etc.

[0052] The synthesis reaction of III-e can be carried out in the presence of a reaction reagent, a catalyst, and an activator in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, ethanol, DME, dichloromethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the catalyst can be selected from, but not limited to, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex, [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium, tetrakis(triphenylphosphine)palladium, methanesulfonic acid (2-dicyclohexylphosphino-2',6'-diisopropyloxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II), methanesulfonic acid(2-dicyclohexylphosphino)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II), bis(triphenylphosphine)palladium(II) chloride, triphenylphosphine gold chloride, etc.; the activator can be selected from but not limited to silver hexafluoroantimonate, silver tetrafluoroborate, etc.

[0053] The synthesis reaction of III-f can be carried out in the presence of a reaction reagent and a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the base can be selected from, but not limited to, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, N,N-diisopropylethylamine, pyridine, and the like.

[0054] The synthesis reaction of III-g can be carried out in the presence of a reaction reagent, a catalyst, and a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the catalyst can be selected from, but not limited to, sodium iodide, potassium iodide, etc.; the base can be selected from, but not limited to, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, N,N-diisopropylethylamine, pyridine, etc.

[0055] The synthesis reaction of III-h and III-h-1 can be carried out in the presence of a reaction reagent, an acid, or a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the acid can be selected from, but not limited to, hydrochloric acid, trifluoroacetic acid, acetic acid, boric acid, sulfuric acid, and the like; and the base can be selected from, but not limited to, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, N,N-diisopropylethylamine, pyridine, and the like.

[0056] The synthesis reaction of III-1 and III-1-bp can be carried out in the presence of a reaction reagent and a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the base can be selected from, but not limited to, N,N-diisopropylethylamine, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, pyridine, and the like.

[0057] Alternatively, compound III-1 may be synthesized using the following method IV:

[0058] In each formula, R1, R2, R3, R4, and R5 are as defined in formula (I).

[0059] The synthesis reaction of IV-b can be carried out in the presence of a reaction reagent and an acid in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the acid can be selected from, but not limited to, hydrochloric acid, trifluoroacetic acid, acetic acid, boric acid, sulfuric acid, aluminum chloride, ferric chloride, and the like.

[0060] The synthesis reaction of IV-c can be carried out in the presence of a reaction reagent and an oxidizing agent in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the oxidizing agent can be selected from, but not limited to, tin dioxide, manganese dioxide, sodium hypochlorite, and the like.

[0061] The synthesis reaction of IV-d can be carried out in the presence of a reaction reagent and a reducing agent in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, methanol, water, or any mixture thereof; the reducing agent can be selected from, but not limited to, sodium borohydride, lithium borohydride, platinum dioxide, and the like.

[0062] The synthesis reaction of IV-e can be carried out in the presence of a reaction reagent and a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the base can be selected from, but not limited to, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, N,N-diisopropylethylamine, pyridine, and the like.

[0063] The synthesis reaction of IV-f can be carried out in the presence of a reaction reagent, a catalyst, and a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the catalyst can be selected from, but not limited to, sodium iodide, potassium iodide, or the like; and the base can be selected from, but not limited to, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, N,N-diisopropylethylamine, pyridine, or the like.

[0064] The synthesis reaction of IV-g can be carried out in the presence of a reaction reagent, an acid, or a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the acid can be selected from, but not limited to, hydrochloric acid, trifluoroacetic acid, acetic acid, boric acid, sulfuric acid, or the like; and the base can be selected from, but not limited to, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, N,N-diisopropylethylamine, pyridine, or the like.

[0065] The synthesis reaction of III-1 can be carried out in the presence of a reaction reagent and a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the base can be selected from, but not limited to, N,N-diisopropylethylamine, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, pyridine, and the like.

[0066] Compound IV-1 was prepared using the following method V:

[0067] In each formula, R1, R2, R3, R4, and R5 are as defined in formula (I).

[0068] The synthesis reaction of Vb can be carried out in the presence of a reaction reagent, an activating agent, and a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the activating agent can be selected from, but not limited to, phosphorus trichloride, phosphorus oxychloride, phosphorus pentachloride, or the like; and the base can be selected from, but not limited to, N,N-diisopropylethylamine, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, pyridine, or the like.

[0069] The synthesis reaction of Vc can be carried out in the presence of a reaction reagent, a catalyst, and a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the catalyst can be selected from, but not limited to, sodium iodide, potassium iodide, etc.; the base can be selected from, but not limited to, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, N,N-diisopropylethylamine, pyridine, etc.

[0070] The synthesis reaction of Vd can be carried out in the presence of a reaction reagent, an acid, or a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the acid can be selected from, but not limited to, hydrochloric acid, trifluoroacetic acid, acetic acid, boric acid, sulfuric acid, or the like; and the base can be selected from, but not limited to, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, N,N-diisopropylethylamine, pyridine, or the like.

[0071] The synthesis reaction of IV-1 can be carried out in the presence of a reaction reagent and a base in a solvent. The solvent can be selected from, but not limited to, tetrahydrofuran, 1,4-dioxane, toluene, xylene, acetonitrile, dichloromethane, 1,2-dichloroethane, dimethyl sulfoxide, N,N-dimethylformamide, water, or any mixture thereof; the base can be selected from, but not limited to, N,N-diisopropylethylamine, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, triethylamine, pyridine, and the like.

[0072] Optionally, the compound represented by formula (I) or its deuterated form, or pharmaceutically acceptable salts thereof, include anionic salts and cationic salts of the compound represented by formula (I) or its deuterated form;

[0073] Optionally, the pharmaceutically acceptable salt includes an alkali metal salt, an alkaline earth metal salt, or an ammonium salt of the compound represented by formula (I) or a deuterated product thereof; optionally, the alkali metal includes sodium, potassium, lithium, and cesium, and the alkaline earth metal includes magnesium, calcium, and strontium;

[0074] Optionally, the pharmaceutically acceptable salt includes a salt formed by the compound represented by formula (I) or a deuterated substance thereof and an organic base;

[0075] Optionally, the organic base includes trialkylamine, pyridine, quinoline, piperidine, imidazole, picoline, dimethylaminopyridine, dimethylaniline, N-alkylmorpholine, 1,5-diazabicyclo[4.3.0]nonene-5, 1,8-diazabicyclo[5.4.0]undecene-7, 1,4-diazabicyclo[2.2.2]octane; Optionally, the trialkylamine includes trimethylamine, triethylamine, N-ethyldiisopropylamine; Optionally, the N-alkylmorpholine includes N-methylmorpholine;

[0076] Optionally, the pharmaceutically acceptable salt includes a salt formed between the compound represented by formula (I) or a deuterated substance thereof and an acid;

[0077] Optionally, the acid includes an inorganic acid or an organic acid; optionally, the inorganic acid includes hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, and carbonic acid; optionally, the organic acid includes formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, citric acid, citric acid, tartaric acid, carbonic acid, picric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, glutamic acid, and pamoic acid.

[0078] In another aspect, the present invention provides a pharmaceutical composition comprising the compound represented by formula (I) or a deuterated substance thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutical excipient.

[0079] The pharmaceutical excipients are conventionally selected in the pharmaceutical field and are not particularly limited in the present invention.

[0080] In another aspect, the present invention provides the use of the compound represented by formula (I) above or its deuterated form, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition thereof in the preparation of an SLC7A11 inhibitor.

[0081] In another aspect, the present invention provides a compound represented by the above formula (I) or a deuterated substance thereof, or a pharmaceutically acceptable salt thereof, or the above pharmaceutical composition for use in inhibiting SLC7A11.

[0082] In another aspect, the present invention provides a method for inhibiting SLC7A11, comprising administering to a subject in need thereof an effective amount of the compound represented by formula (I) above or its deuterated form, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described above.

[0083] Another aspect of the present invention provides the use of the compound represented by formula (I), a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described above in the preparation of a drug for treating tumor diseases such as pancreatic cancer, liver cancer, osteosarcoma, and glioma.

[0084] In another aspect, the present invention provides a compound represented by the above formula (I) or a deuterated product thereof, or a pharmaceutically acceptable salt thereof, or the above pharmaceutical composition for use in treating tumor diseases such as pancreatic cancer, liver cancer, osteosarcoma, glioma, etc.

[0085] In another aspect, the present invention provides a method for treating pancreatic cancer, liver cancer, osteosarcoma, glioma and other tumor diseases, comprising administering to a subject in need thereof an effective amount of the compound represented by the above formula (I) or its deuterated form, or their pharmaceutically acceptable salts, or the above pharmaceutical composition.

[0086] In another aspect, the present invention provides the use of the compound represented by the above formula (I), a pharmaceutically acceptable salt thereof, or the above pharmaceutical composition in the preparation of a drug for treating chronic diseases such as fatty liver, cirrhosis, liver fibrosis, pulmonary fibrosis, diabetic skin lesions, or neurodegenerative diseases such as amyotrophic lateral sclerosis, Parkinson's disease, and Alzheimer's disease.

[0087] In another aspect, the present invention provides a compound represented by the above formula (I) or a deuterated product thereof, or a pharmaceutically acceptable salt thereof, or the above pharmaceutical composition, for use in treating chronic diseases such as fatty liver, cirrhosis, liver fibrosis, pulmonary fibrosis, diabetic skin lesions, or neurodegenerative diseases such as amyotrophic lateral sclerosis, Parkinson's disease, and Alzheimer's disease.

[0088] In another aspect, the present invention provides a method for treating chronic diseases such as fatty liver, cirrhosis, liver fibrosis, pulmonary fibrosis, diabetic skin lesions, or neurodegenerative diseases such as amyotrophic lateral sclerosis, Parkinson's disease, and Alzheimer's disease, comprising administering to a subject in need thereof an effective amount of the compound represented by formula (I) above or its deuterated form, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described above.

[0089] Beneficial effects of the present invention:

[0090] The compounds obtained in the present invention can effectively inhibit the activity of SLC7A11 and can be used to prevent and / or treat diseases related to SLC7A11. BRIEF DESCRIPTION OF THE DRAWINGS

[0091] FIG1 shows the effect of compounds (including compound 186, compound 199 / 200, compound 184 / 185) on promoting BMDC cell efferocytosis. DETAILED DESCRIPTION

[0092] The present invention is further described by the following examples. It should be understood that the examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention in any way.

[0093] Example 1

[0094] Synthesis of 1-fluoro-5-isopropoxy-4-nitro-2-(trifluoromethyl)benzene:

[0095] Sodium metal (66 mg, 2.89 mmol) was added to isopropanol (10 mL), and the mixture was heated at 80 ° C for 2 hours. The mixture was added dropwise to a solution of 1,5-difluoro-2-nitro-4-(trifluoromethyl)benzene (600 mg, 2.63 mmol) in tetrahydrofuran (20 mL), and the mixture was stirred at room temperature for 16 hours. The mixture was concentrated in vacuo, and the residue was diluted with ethyl acetate (30 mL) and washed with saturated brine (30 mL). It was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a yellow oily crude product 1-3 (650 mg, 88%).

[0096] LC-MS: [M+H] + :No MS.

[0097] 1 H NMR: (400MHz, CDCl3) δ8.16 (d, J = 7.6Hz, 1H), 6.87 (d, J = 12.0Hz, 1H), 4.72-4.66 (m, 1H), 1.44 (d, J = 6.0Hz, 6H).

[0098] Synthesis of 4-fluoro-2-isopropoxy-5-(trifluoromethyl)aniline:

[0099] To a solution of 1-fluoro-5-isopropoxy-4-nitro-2-(trifluoromethyl)benzene 1-3 (650 mg, 2.42 mmol) in methanol (20 mL) was added 20% palladium hydroxide on carbon (68 mg, 0.48 mmol), and the mixture was hydrogenated with a hydrogen balloon for 3 hours. The mixture was filtered and the filtrate was concentrated in vacuo to give a black oily crude product 1-4 (550 mg, 93%).

[0100] LC-MS: [M+H] + :238.1.

[0101] Synthesis of 2-(chloromethyl)-3-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)quinazolin-4(3H)-one:

[0102] To a solution of 2-(2-chloroacetamido)benzoic acid 1-5 (90 mg, 0.42 mmol, doi.org / 10.1016 / j.bmcl.2011.05.031) in acetonitrile (10 mL) were added N,N-diisopropylethylamine (54 mg, 0.42 mmol) and phosphorus trichloride (116 mg, 0.84 mmol), and the mixture was stirred at room temperature for 10 minutes. A solution of 4-fluoro-2-isopropoxy-5-(trifluoromethyl)aniline 1-4 (100 mg, 0.42 mmol) in acetonitrile (0.5 mL) was then added, and the mixture was stirred at 70°C for 5 hours. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with ethyl acetate (60 mL) and washed with saturated brine (30 mL x 2). The separated organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether = 1 / 2) to give a white solid product 1-6 (160 mg, 87%)

[0103] LC-MS: [M+H] + :415.1.

[0104] Synthesis of (S)-tert-butyl 4-((3-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3-methylpiperazine-1-carboxylate:

[0105] A mixture of 2-(chloromethyl)-3-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)quinazoline-4(3H)-one 1-6 (100 mg, 0.24 mmol), (S)-3-methylpiperazine-1-carboxylic acid tert-butyl ester 1-7 (48 mg, 0.24 mmol), potassium carbonate (50 mg, 0.36 mmol) and potassium iodide (40 mg, 0.24 mmol) in acetonitrile (20 mL) was heated at 80°C for 16 hours. The mixture was concentrated in vacuo, and the residue was diluted with ethyl acetate (30 mL) and washed with saturated brine (20 mL). The separated organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether = 1 / 3) to give a yellow solid product 1-8 (100 mg, 68%).

[0106] LC-MS: [M+H] + :579.3.

[0107] Synthesis of (S)-3-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)-2-((2-methylpiperazin-1-yl)methyl)quinazolin-4(3H)-one:

[0108] To a solution of (S)-tert-butyl 4-((3-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3-methylpiperazine-1-carboxylate 1-8 (100 mg, 0.17 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (2 mL), and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure to give the crude product 1-9 (80 mg, 92%) as a yellow oil.

[0109] LC-MS: [M+H] + :479.2.

[0110] Synthesis of (S)-2-((4-(2-(4-chlorophenoxy)acetyl)-2-methylpiperazin-1-yl)methyl)-3-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)quinazolin-4(3H)-one:

[0111] To a solution of (S)-3-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)-2-((2-methylpiperazin-1-yl)methyl)quinazolin-4(3H)-one 1-9 (80 mg, 0.17 mmol) and N,N-diisopropylethylamine (64.7 mg, 0.50 mmol) in dichloromethane (10 mL) was added 2-(4-chlorophenoxy)acetyl chloride (41 mg, 0.20 mmol) at 0°C, and the mixture was stirred at room temperature for 2 hours. The mixture was diluted with dichloromethane (50 mL) and washed with saturated brine (30 mL×2). The separated organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by preparative high performance chromatography (acetonitrile-water 0.1% formic acid, gradient 40%-80%) to give compound 136 (60 mg, 82%) of Example 1 as a white solid product.

[0112] LC-MS: [M+H] + :647.3.

[0113] 1H NMR: (400MHz, CD3OD) δ8.13(d,J=7.2Hz,1H),7.82-7.73(m,2H),7.65(d,J=8.0Hz,1H) ,7.50(t,J=7.6Hz,1H),7.22-7.13(m,3H),6.84-6.81(m,2H),4.70-4.59(m,3H),3.76- 3.64(m,2H),3.47-3.29(m,2H),3.10-3.05(m,1H),2.94-2.79(m,1H),2.62-2.53(m,1 H),2.30-2.10(m,2H),1.15(d,J=6.0Hz,3H),1.10(d,J=6.0Hz,3H),0.68-0.60(m,3H).

[0114] Example 2 / 3

[0115] Compound 136 (50 mg) of Example 1 was subjected to chiral separation by SFC (chromatographic column: CHIRALPAK WHELK-O1 250 mm × 21.1 mm, 5 μm, mobile phase: carbon dioxide-ethanol (NH4OH 0.2%) gradient: 0-40%) to give the title compound (21.1 mg, 23 mg)

[0116] Single configuration compound (shorter retention time)

[0117] It was analyzed by chiral analysis chromatography conditions, RT = 8.88min

[0118] LC-MS: [M+H] + :647.2.

[0119] HPLC: 96.63% [214 nm].

[0120] 1H NMR: (400MHz, CD3OD) δ8.13(d,J=7.2Hz,1H),7.82-7.73(m,2H),7.65(d,J=8.0Hz,1H) ,7.50(t,J=7.6Hz,1H),7.22-7.13(m,3H),6.84-6.81(m,2H),4.70-4.59(m,3H),3.76- 3.64(m,2H),3.47-3.29(m,2H),3.10-3.05(m,1H),2.94-2.79(m,1H),2.62-2.53(m,1 H),2.30-2.10(m,2H),1.15(d,J=6.0Hz,3H),1.10(d,J=6.0Hz,3H),0.68-0.60(m,3H).

[0121] Single configuration compound (longer retention time)

[0122] It was analyzed by chiral analysis chromatography conditions, RT = 10.33min

[0123] LC-MS: [M+H] + :647.1.

[0124] HPLC: 98.34% [214 nm].

[0125] 1 H NMR: (400MHz, CD3OD) δ8.13(d,J=7.2Hz,1H),7.82-7.73(m,2H),7.65(d,J=8.0Hz,1H) ,7.50(t,J=7.6Hz,1H),7.22-7.13(m,3H),6.84-6.81(m,2H),4.70-4.59(m,3H),3.76- 3.64(m,2H),3.47-3.29(m,2H),3.10-3.05(m,1H),2.94-2.79(m,1H),2.62-2.53(m,1 H),2.30-2.10(m,2H),1.15(d,J=6.0Hz,3H),1.10(d,J=6.0Hz,3H),0.68-0.60(m,3H).

[0126] Example 4

[0127] Synthesis of 1-cyclopropyloxy-2-nitro-4-(trifluoromethyl)benzene:

[0128] To a solution of 1-fluoro-2-nitro-4-(trifluoromethyl)benzene 4-1 (100.0 mg, 0.48 mmol) in N,N-dimethylformamide (10 mL) was added potassium carbonate (187.0 mg, 0.57 mmol). After stirring for 15 minutes, cyclopropanol 4-2 (27.8 mg, 0.48 mmol) was added and the mixture was stirred at 70°C for 16 hours. After dilution with water (30 mL), the mixture was extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with saturated sodium chloride solution (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether = 1 / 1) to give the product 4-3 (70 mg, 57%) as a white solid.

[0129] 1 H NMR: (400MHz, DMSO-d6) δ8.31(d,J=2.0Hz,1H),8.09(dd,J=8.9,2.1Hz,1H),7.83(d,J=8.8Hz,1H),4.22(dt,J=9.0,3.0Hz,1H),0.92 -0.88(m,2H),0.80 -0.76(m,2H).

[0130] Synthesis of 2-cyclopropyloxy-5-(trifluoromethyl)aniline:

[0131] To a solution of 1-cyclopropyloxy-2-nitro-4-(trifluoromethyl)benzene 4-3 (50 mg, 0.20 mmol) in ethanol (20 mL) was added saturated aqueous ammonium chloride (1 mL) and iron powder (45 mg, 0.80 mmol), and the mixture was reacted at 80°C for 16 hours. The mixture was then filtered, and the filtrate was concentrated under reduced pressure. The residue was diluted with saturated brine (20 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The filtrate was then dried under vacuum to give the product 4-4 as a white solid (45 mg crude).

[0132] LC-MS: [M+H] + :218.1.

[0133] According to steps 3, 4, 5 and 6 in Example 1, replacing raw material 1-4 with 4-4 can obtain Example 4(S)-2-((4-(2-(4-chlorophenoxy)acetyl)-2-methylpiperazin-1-yl)methyl)-3-(2-cyclopropyloxy-5-(trifluoromethyl)phenyl)quinazoline-4(3H)-one (39.5 mg).

[0134] LC-MS: [M+H]+: 627.0.

[0135] HPLC: 99.37%.

[0136] 1 H NMR: (400MHz, CD3OD) δ8.24 (d, J=8.0Hz, 1H), 7.93-7.84 (m, 3H), 7.77-7.74 (m, 2H),7.61(t,J=7.6Hz,1H),7.30-7.25(m,2H),6.95-6.92(m,2H),4.80-4.71(m ,2H),4.03-3.91(m,2H),3.62-3.51(m,2H),3.25-3.20(m,1H),3.05-2.97(m,1 H),2.81-2.69(m,2H),2.28-2.20(m,2H),0.87-0.81(m,2H),0.69-0.56(m,5H).

[0137] Example 5 / 6

[0138] Compound 3 (37.5 mg) from Example 4 was subjected to chiral separation by SFC (column: CHIRALPAK OJ-H 250 mm x 20 mm, 5 μm, mobile phase: carbon dioxide-methanol (NH4OH 0.2%) gradient: 0-40%) to give the title compound (13.1 mg, 14.1 mg).

[0139] Single configuration compound (shorter retention time)

[0140] It was analyzed by chiral analysis chromatography conditions, RT = 2.19min

[0141] LC-MS: [M+H]+: 627.0.

[0142] HPLC: 100%.

[0143] 1 H NMR: (400MHz, CD3OD) δ8.24 (d, J=8.0Hz, 1H), 7.93-7.84 (m, 3H), 7.77-7.74 (m, 2H),7.61(t,J=7.6Hz,1H),7.30-7.25(m,2H),6.95-6.92(m,2H),4.80-4.71(m ,2H),4.03-3.91(m,2H),3.62-3.51(m,2H),3.25-3.20(m,1H),3.05-2.97(m,1 H),2.81-2.69(m,2H),2.28-2.20(m,2H),0.87-0.81(m,2H),0.69-0.56(m,5H).

[0144] Single configuration compound (longer retention time)

[0145] It was analyzed by chiral analysis chromatography conditions, RT = 2.54min

[0146] LC-MS: [M+H]+: 627.0.

[0147] HPLC: 100%.

[0148] 1 H NMR: (400MHz, CD3OD) δ8.24 (d, J=8.0Hz, 1H), 7.93-7.84 (m, 3H), 7.77-7.74 (m, 2H),7.61(t,J=7.6Hz,1H),7.30-7.25(m,2H),6.95-6.92(m,2H),4.80-4.71(m ,2H),4.03-3.91(m,2H),3.62-3.51(m,2H),3.25-3.20(m,1H),3.05-2.97(m,1 H),2.81-2.69(m,2H),2.28-2.20(m,2H),0.87-0.81(m,2H),0.69-0.56(m,5H).

[0149] Example 7

[0150] Referring to Example 1, replacing 1-7 with 7-1 can give (S)-2-((4-(2-(4-chlorophenoxy)acetyl)-2-ethylpiperazin-1-yl)methyl)-3-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)quinazolin-4(3H)-one compound 139 (55.8 mg, 34.9%).

[0151] LC-MS: [M+H]+: 661.2.

[0152] HPLC: 99.2%

[0153] 1HNMR(400MHz,CD3OD)δ8.20(d,J=8.0Hz,1H),7.90-7.79(m,2H),7.74-7.72(m,1H),7 .55(t,J=7.6Hz,1H),7.31-7.19(m,3H),6.93-6.86(m,2H),4.82-4.63(m,3H),3.80-3 .31(m,4H),3.24-2.91(m,2H),2.76-2.56(m,1H),2.43-1.90(m,2H),1.43-1.25(m,1H ),1.23(d,J=6.0Hz,3H),1.17(d,J=6.0Hz,3H),1.05-0.81(m,1H),0.78-0.68(m,3H).

[0154] Example 8

[0155] Referring to Example 1, by replacing 1-7 with 8-1, (S)-2-((4-(2-(4-chlorophenoxy)acetyl)-2-isopropylpiperazin-1-yl)methyl)-3-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)quinazolin-4(3H)-one compound 141 (52.8 mg, 30.5%) can be obtained.

[0156] LC-MS: [M+H] + :675.3.

[0157] HPLC: 99.8%

[0158] 1 HNMR(400MHz,CD3OD)δ8.19(d,J=8.0Hz,1H),7.70-7.52(m,4H),7.33-7.18(m,3H),6. 94-6.88(m,2H),4.82-4.62(m,3H),4.18-4.00(m,1H),3.88-3.60(m,2H),3.45-3.31(m ,2H),3.19-2.18(m,5H),2.02-1.74(m,1H),1.25(d,J=6.0Hz,3H),1.16(d,J=6.0Hz,3 H),0.87-0.82(m,2H),0.74(d,J=6.8Hz,1H),0.65-0.60(m,1H),0.45(t,J=6.8Hz,2H).

[0159] Example 9

[0160] Referring to Example 1, by replacing 1-7 with 9-1, 2-((4-(2-(4-chlorophenoxy)acetyl)-2-cyclopropylpiperazin-1-yl)methyl)-3-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)quinazolin-4(3H)-one, compound 142 (35.1 mg, 29.0%) can be obtained.

[0161] LC-MS: [M+H]+: 673.3.

[0162] HPLC: 98.5%

[0163] 1 H NMR(400MHz,CD3OD)δ8.03(d,J=7.8Hz,1H),7.73-7.64(m,2H),7.58-7.50(m,1H),7.43-7.36(m ,1H),7.12-7.02(m,3H),6.71(dd,J=8.8Hz,2.0Hz,2H),4.61-4.47(m,3H),4.34-4.31(m,1H),4. 14-3.78(m,2H),3.53-3.46(m,1H),3.05-2.62(m,3H),2.06-1.96(m,1H),1.57-1.21(m,1H),1.0 9-1.04(m,3H),1.04-0.95(m,3H),0.78(t,J=7.4Hz,1H),0.39-0.11(m,2H),0.05--0.21(m,2H).

[0164] Example 10

[0165] Synthesis of 3-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)-2-(piperazin-1-ylmethyl)quinazolin-4(3H)-one:

[0166] A mixture of 2-(chloromethyl)-3-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)quinazolin-4(3H)-one 1-6 (90 mg, 0.22 mmol) and piperazine 10-1 (81 mg, 0.43 mmol) in tetrahydrofuran (20 mL) was heated at 70°C for 16 hours. The mixture was concentrated in vacuo, and the residue was diluted with ethyl acetate (30 mL) and washed with saturated brine (20 mL). The separated organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the product 10-2 (100 mg) as a yellow solid.

[0167] LC-MS: [M+H] + :465.2.

[0168] Synthesis of (2-((4-(2-(4-chlorophenoxy)acetyl)piperazin-1-yl)methyl)-3-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)quinazolin-4(3H)-one:

[0169] To a solution of 3-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)-2-(piperazine-1-ylmethyl)quinazoline-4(3H)-one 10-2 (100 mg, 0.22 mmol) and N,N-diisopropylethylamine (111.1 mg, 0.86 mmol) in dichloromethane (10 mL) was added 2-(4-chlorophenoxy)acetyl chloride 1-10 (41 mg, 0.20 mmol) at 0°C, and the mixture was stirred at room temperature for 2 hours. The mixture was diluted with dichloromethane (50 mL) and washed with saturated brine (30 mL×2). The separated organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by preparative high performance chromatography (acetonitrile-water 0.1% formic acid, gradient 40%-80%) to give compound 14 (115 mg, 84.56%) as a white solid product.

[0170] LC-MS: [M+H] + :633.1.

[0171] HPLC: 98.63% [214 nm].

[0172] 1 H NMR: (400MHz, CD3OD) δ8.26(dd,J=8.0,1.2Hz,1H),7.96-7.88(m,2H),7.81(d,J=8.0Hz,1H),7.63(t,J=7.2Hz,1H),7.23-7.12( m,3H),6.98-6.91(m,2H),4.84-4.75(m,3H),3.36-3.20(m,6H),2.53-2.30(m,4H),1.27(d,J=6.0Hz,3H),1.21(d,J=6.0Hz,3H).

[0173] Example 11 / 12

[0174] Compound 14 (81 mg) was chiral separated by SFC (chromatographic column: CHIRALPAK AD-H 250 mm 20 mm, 5 μm mobile phase: carbon dioxide-isopropanol (NH4OH 0.2%) gradient: 0-40%) to obtain the title compound (36 mg, 40 mg)

[0175] Single configuration compound (shorter retention time)

[0176] It was analyzed by chiral analysis chromatography conditions, RT = 2.04min

[0177] LC-MS: [M+H] + :633.2.

[0178] HPLC: 98.71% [214 nm].

[0179] 1 H NMR: (400MHz, CD3OD) δ8.26(dd,J=8.0,1.2Hz,1H),7.96-7.88(m,2H),7.81(d,J=8.0Hz,1H),7.63(t,J=7.2Hz,1H),7.23-7.12( m,3H),6.98-6.91(m,2H),4.84-4.75(m,3H),3.36-3.20(m,6H),2.53-2.30(m,4H),1.27(d,J=6.0Hz,3H),1.21(d,J=6.0Hz,3H).

[0180] Single configuration compound (longer retention time)

[0181] It was analyzed by chiral analysis chromatography conditions, RT = 2.93min

[0182] LC-MS: [M+H] + :633.1.

[0183] HPLC: 99.44% [214 nm].

[0184] 1 H NMR: (400MHz, CD3OD) δ8.26(dd,J=8.0,1.2Hz,1H),7.96-7.88(m,2H),7.81(d,J=8.0Hz,1H),7.63(t,J=7.2Hz,1H),7.23-7.12( m,3H),6.98-6.91(m,2H),4.84-4.75(m,3H),3.36-3.20(m,6H),2.53-2.30(m,4H),1.27(d,J=6.0Hz,3H),1.21(d,J=6.0Hz,3H).

[0185] Example 13

[0186] Synthesis of 2-bromo-6-isopropoxy-3-(trifluoromethyl)aniline:

[0187] To a solution of 2-isopropoxy-5-(trifluoromethyl)aniline 13-1 (500 mg, 2.28 mmol) in carbon tetrachloride (20 mL) at -10 ° C, N-bromosuccinimide (406 mg, 2.28 mmol) was slowly added, and the mixture was stirred at 0 ° C for 2 hours. The mixture was quenched with an aqueous sodium sulfite solution (20 mL) and then extracted with dichloromethane (20 mL x 3). The combined organic layer was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether=1 / 5) to give a yellow oily product 13-2 (550 mg, 73%).

[0188] LC-MS: [M+H] + :298.0,300.0.

[0189] 1 H NMR: (400MHz, CDCl3) δ7.04 (d, J = 8.8Hz, 1H), 6.71 (d, J = 8.8Hz, 1H), 4.65-4.58 (m, 1H), 1.38 (d, J = 6.0Hz, 6H).

[0190] Synthesis of 6-isopropoxy-2-methyl-3-(trifluoromethyl)aniline:

[0191] To a solution of 2-bromo-6-isopropoxy-3-(trifluoromethyl)aniline 13-2 (200 mg, 0.67 mmol), 2,4,6-trimethyl-1,3,5,2,4,6-trimethoxytriborane 13-3 (168 mg, 1.34 mmol), and potassium carbonate (185 mg, 1.34 mmol) in N,N-dimethylformamide (20 mL) was added 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (55 mg, 0.067 mmol), and the mixture was heated at 110°C for 16 hours. After cooling to room temperature, the mixture was diluted with water (30 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether = 1 / 6) to give a white solid product 13-4 (110 mg, 67%).

[0192] LC-MS: [M+H] + :234.2.

[0193] 1H NMR: (400MHz, CDCl3) δ7.17 (d, J = 8.8Hz, 1H), 6.72 (d, J = 8.8Hz, 1H), 4.67-4.61 (m, 1H), 2.38 (s, 3H), 1.39 (d, J = 6.0Hz, 6H).

[0194] Referring to step 3 in Example 1 and steps 1 and 2 in Example 10, replacing 1-4 with 13-4 can give 2-((4-(2-(4-chlorophenoxy)acetyl)piperazin-1-yl)methyl)-3-(6-isopropoxy-2-methyl-3-(trifluoromethyl)phenyl)quinazolin-4(3H)-one, compound 75 (90 mg, 57%).

[0195] LC-MS: [M+H] + :629.2.

[0196] HPLC: 95.37% [214 nm].

[0197] 1 H NMR: (400MHz, DMSO-d6)δ8.16(dd,J=8.0,1.2Hz,1H),7.93-7.88(m,1H),7.79( dd,J=12.4,8.4Hz,2H),7.64-7.58(m,1H),7.33-7.25(m,3H),6.92-6.87(m,2H) ,4.82-4.72(m,3H),3.29-3.18(m,4H),3.15(d,J=6.4Hz,2H),2.46-2.32(m,2H ),2.20(s,3H),1.92-1.81(s,2H),1.16(d,J=6.0Hz,3H),1.06(d,J=6.0Hz,3H).

[0198] Example 14 / 15

[0199] Compound 75 (80 mg) was chiral separated by SFC (chromatographic column: AD-H 250 mm × 20 mm, 5 μm, mobile phase: carbon dioxide-ethanol (NH4OH 0.2%) gradient: 0-40%) to obtain the title compound (23.5 mg, 24.1 mg)

[0200] Single configuration compound (shorter retention time)

[0201] It was analyzed by chiral analysis chromatography conditions, RT = 2.52min

[0202] LC-MS: [M+H] + :629.3.

[0203] HPLC: 96.41% [214 nm].

[0204] 1 H NMR: (400MHz, DMSO-d6)δ8.16(d,J=7.6Hz,1H),7.91(t,J=7.6Hz,2H),7.79(dd, J=12.4,8.4Hz,2H),7.60(t,J=7.6Hz,1H),7.31-7.25(m,3H),6.90(d,J=8.8Hz,2 H),4.81-4.73(m,3H),3.27-3.15(m,4H),3.15(d,J=6.0Hz,2H),2.42-2.34(m,2 H),2.20(s,3H),1.91-1.80(m,2H),1.16(d,J=6.0Hz,3H),1.07(d,J=6.0Hz,3H).

[0205] Single configuration compound (longer retention time)

[0206] It was analyzed by chiral analysis chromatography conditions, RT = 3.00 min

[0207] LC-MS: [M+H] + :629.3.

[0208] HPLC: 99.03% [214 nm].

[0209] 1 H NMR: (400MHz, DMSO-d6)δ8.16(d,J=7.6Hz,1H),7.91(t,J=7.6Hz,1H),7.79(dd, J=12.4,8.8Hz,2H),7.60(t,J=7.6Hz,1H),7.31-7.26(m,3H),6.90(d,J=8.8Hz,2 H),4.84-4.71(m,3H),3.28-3.16(m,4H),3.15(d,J=6.0Hz,2H),2.44-2.33(m,2 H),2.20(s,3H),1.92-1.80(m,2H),1.16(d,J=6.0Hz,3H),1.07(d,J=6.0Hz,3H).

[0210] Example 16

[0211] Synthesis of 6-isopropoxy-2-(prop-1-en-2-yl)-3-(trifluoromethyl)aniline:

[0212] To a solution of 2-bromo-6-isopropoxy-3-(trifluoromethyl)aniline 13-2 (150 mg, 0.5 mmol), 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane 14-1 (101 mg, 0.6 mmol), and sodium carbonate (107 mg, 1 mmol) in dioxane (15 mL) and water (3 mL) was added 1,1-bis(diphenylphosphino)ferrocenepalladium dichloride dichloromethane complex (37 mg, 0.05 mmol), and the mixture was heated at 80°C for 16 hours. The mixture was diluted with water (20 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether = 1 / 8) to give a yellow oily product 14-2 (100 mg, 72.8%).

[0213] LC-MS: [M+H] + :260.1.

[0214] Synthesis of 6-isopropoxy-2-isopropyl-3-(trifluoromethyl)aniline:

[0215] A mixture of 6-isopropoxy-2-(prop-1-en-2-yl)-3-(trifluoromethyl)aniline 14-2 (100 mg, 0.39 mmol) and palladium hydroxide on carbon (54 mg) in methanol (20 mL) was hydrogenated with a hydrogen balloon for 16 hours. The mixture was filtered and the filtrate was concentrated in vacuo to give the crude product 14-3 (100 mg, 75%) as a yellow oil.

[0216] LC-MS: [M+H] + :262.1.

[0217] Referring to step 3 in Example 1 and steps 1 and 2 in Example 10, replacing 1-4 with 14-3 can give 2-((4-(2-(4-chlorophenoxy)acetyl)piperazin-1-yl)methyl)-3-(6-isopropoxy-2-isopropyl-3-(trifluoromethyl)phenyl)quinazolin-4(3H)-one, compound 78 (13 mg, 19%).

[0218] LC-MS: [M+H] + :657.3.

[0219] HPLC: 98.85% [214 nm].

[0220] 1H NMR: (400MHz, CD3OD) δ8.20(d,J=8.0Hz,1H),7.92-7.85(m,2H),7.78(d,J=8.0Hz,1H),7.58(t,J=7.6Hz,1H),7.22-7.20(m,3H),6.91 (d,J=8.8Hz,2H),4.77-4.69(m,3H),3.62-3.52(m,4H),3.42-3.32(m,1H),3.28-3.24(m,2H),2.65-2.41(m,4H),1.21-1.09(m,12H).

[0221] Example 17

[0222] Synthesis of 4-bromo-2-isopropoxy-5-(trifluoromethyl)aniline:

[0223] To a solution of 2-isopropoxy-5-(trifluoromethyl)aniline 13-1 (554 mg, 2.5 mmol) in anhydrous acetonitrile (10 mL) at room temperature was added N-bromosuccinimide (448 mg, 2.5 mmol), and the mixture was stirred at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure in vacuo and diluted with ethyl acetate (100 mL), washed with water (100 mL x 2) and saturated brine (100 mL) in sequence. The separated organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified on a silica gel column (petroleum ether: ethyl acetate = 5:1) to obtain the black oily product 17-1 (520 mg, 69%).

[0224] LC-MS: [M+H] + :298.1.

[0225] 1 HNMR (400MHz, CDCl3) δ7.11 (s, 1H), 7.03 (s, 1H), 4.59 (dt, J = 12.0, 6.0Hz, 1H), 1.38 (d, J = 6.0Hz, 6H).

[0226] Synthesis of 2-isopropoxy-4-methyl-5-(trifluoromethyl)aniline:

[0227] To a solution of 4-bromo-2-isopropoxy-5-(trifluoromethyl)aniline 17-1 (100 mg, 0.33 mmol) in dioxane / water (10 / 5 mL) was added 2,4,6-trimethyl-1,3,5,2,4,6-trioxytriborane 13-3 (0.4 mL), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex (24.5 mg, 0.033 mmol), and sodium carbonate (71 mg, 0.67 mmol) at room temperature. The mixture was heated to 80°C under nitrogen and stirred for 16 hours. The reaction mixture was concentrated under reduced pressure and diluted with ethyl acetate (50 mL). The mixture was washed sequentially with water (100 mL) and saturated brine (100 mL). The separated organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate=3:1) to give a yellow oily product 17-2 (60 mg, 76.5%).

[0228] LC-MS: [M+H] + :234.2.

[0229] Referring to step 3 in Example 1 and steps 1 and 2 in Example 10, replacing 1-4 with 17-2 can give 2-((4-(2-(4-chlorophenoxy)acetyl)piperazin-1-yl)methyl)-3-(2-isopropoxy-4-methyl-5-(trifluoromethyl)phenyl)quinazolin-4(3H)-one, compound 17 (16.5 mg, 20%).

[0230] LC-MS: [M+H] + :628.9.

[0231] HPLC: 98.2%

[0232] 1 H NMR (400MHz, CD3OD) δ8.24(d,J=8.0Hz,1H),7.84-7.82(m,2H),7.72(s,1H),7.62(t,J=7.2Hz,1H),7.28-7.22(m,3H),6.93-6.90(m,2H ),4.82-4.75(m,3H),3.98-3.94(m,1H)3.86-3.44(m,5H),3.25-2.88(m,4H),2.57(s,3H),1.25(d,J=6.0Hz,3H),1.15(d,J=6.0Hz,3H).

[0233] Example 18

[0234] Referring to step 3 in Example 1 and steps 1 and 2 in Example 10, replacing 1-4 with 13-1 can give 2-((4-(2-(4-chlorophenoxy)acetyl)piperazin-1-yl)methyl)-3-(2-isopropoxy-5-(trifluoromethyl)phenyl)quinazolin-4(3H)-one, compound 1 (46.1 mg, 18%).

[0235] LC-MS: [M+H] + :615.2.

[0236] HPLC: 100%

[0237] 1 HNMR (400MHz, CD3OD) δ8.25(d,J=7.9Hz,1H),7.96-7.85(m,3H),7.81(s,1H),7.63(t,J=7.5Hz,1H),7.46(d,J=8.9Hz,1H),7.27-7.22(m,2H) ,6.97-6.91(m,2H),4.86-4.79(m,4H),4.32-4.27(m,1H),3.95-3.91( m,4H),3.56-3.34(m,4H),1.26(d,J=6.0Hz,3H),1.16(d,J=6.0Hz,3H).

[0238] Example 19

[0239] Synthesis of N-(5-acetyl-2-isopropoxyphenyl)-2-iodobenzamide:

[0240] To a solution of 2-iodobenzoic acid 19-1 (250 mg, 1 mmol), 1-(3-amino-4-isopropoxyphenyl)ethanone 19-2 (195 mg, 1 mol), N,N-diisopropylethylamine (391 mg, 3.02 mmol) and 1-hydroxy-7-azabenzotriazole (163 mg, 1.21 mmol) in dichloromethane (20 mL) was added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (290 mg, 1.51 mmol). The mixture was diluted with dichloromethane (30 mL) and washed with saturated brine (30 mL × 3). The separated organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether = 1 / 1) to give a yellow solid product 19-3 (350 mg, 74%).

[0241] LC-MS: [M+H] + :424.0.

[0242] Synthesis of N-(5-acetyl-2-isopropoxyphenyl)-2-(3-hydroxypropyl-1-yl)benzamide:

[0243] A mixture of N-(5-acetyl-2-isopropoxyphenyl)-2-iodobenzamide 19-3 (350 mg, 0.83 mmol), prop-2-yn-1-ol 19-4 (93 mg, 1.65 mmol), CuI (32 mg, 0.16 mmol) and bis(triphenylphosphine)palladium(II) chloride (58 mg, 0.083 mmol) in triethylamine (20 mL) was heated at 35 ° C for 17 hours. After cooling to room temperature, the mixture was concentrated in vacuo, diluted with water (30 mL), and extracted with ethyl acetate (30 mL x 3). The combined organic layer was washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether=1 / 1) to give the product 19-5 (350 mg, 96% yield) as a slightly yellow solid.

[0244] LC-MS: [M+H] + :352.2.

[0245] 1 HNMR: (400MHz, CDCl3) δ9.25 (d, J=2.0Hz, 1H), 9.18 (s, 1H), 7.96-7.93 (m, 1H), 7.77 (dd, J=8.8, 2.4Hz, 1H), 7.57-7.54 (m,1H),7.49-7.46(m,2H),7.01(d,J=8.8Hz,1H),4.72-4.67(m,1H),4.47(s,2H),2.62(s,3H),1.40(d,J=6.0Hz,6H).

[0246] Synthesis of (Z)-1-(3-((3-(hydroxymethyl)-1H-isochrome-1-ylidene)amino)-4-isopropoxyphenyl)ethan-1-one:

[0247] To a solution of silver hexafluoroantimonate (17 mg, 0.050 mmol) and (triphenylphosphine)gold chloride (25 mg, 0.050 mmol) in dichloromethane (10 mL) and dioxane (10 mL) was added N-(5-acetyl-2-isopropoxyphenyl)-2-(3-hydroxypropyl-1-yl)benzamide 19-5 (350 mg, 0.99 mmol), and the mixture was stirred at room temperature for 16 hours. The mixture was concentrated in vacuo, and the residue was diluted with dichloromethane (60 mL) and washed with saturated brine (40 mL). The separated organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether = 1 / 1) to give 19-6 (200 mg, 45.7%) as a yellow solid.

[0248] LC-MS: [M+H] + :352.1.

[0249] 1 HNMR: (400MHz, CDCl3) δ9.16 (s, 1H), 7.99 (s, 1H), 7.87 (d, J = 8.4Hz, 1H), 7.78-7.72 (m, 1H), 7.64-7.58 (m, 1H), 7.45 (d ,J=6.8Hz,1H),7.02(d,J=8.8Hz,1H),6.67(s,1H),4.67-4.60(m,1H),4.38(s,2H),2.54(s,3H),1.26(d,J=6.0Hz,6H).

[0250] Synthesis of (Z)-1-(3-((3-(chloromethyl)-1H-isochrome-1-ylidene)amino)-4-isopropoxyphenyl)ethan-1-one:

[0251] To a solution of (Z)-1-(3-((3-(hydroxymethyl)-1H-isochrom-1-ylidene)amino)-4-isopropoxyphenyl)ethane-1-one 19-6 (200 mg, 0.57 mmol) and N,N-diisopropylethylamine (147 mg, 1.13 mmol) in dichloromethane (20 mL) at 0° C., methylsulfonyl chloride (78 mg, 0.68 mmol) was added, and the mixture was stirred at room temperature for 16 hours. The mixture was diluted with dichloromethane (30 mL) and washed with saturated brine (30 mL). The separated organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether=1 / 3) to give the yellow solid product 19-7 (150 mg, 58.29%).

[0252] LC-MS: [M+H] + :370.1.

[0253] 1 HNMR: (400MHz, CDCl3) δ8.63 (s, 1H), 7.82-7.74 (m, 2H), 7.62 (t, J = 7.6Hz, 1H), 7.53 (t, J = 7.6Hz, 1H), 7.33 (d, J = 7.6Hz, 1H),6.97(d,J=8.4Hz,1H),6.42(s,1H),4.67-4.60(m,1H),4.15(s,2H),2.58-2.52(m,3H),1.29(dd,J=6.0,0.4Hz,6H).

[0254] Synthesis of tert-butyl (Z)-4-((1-((5-acetyl-2-isopropoxyphenyl)imino)-1H-isochrome-3-yl)methyl)piperazine-1-carboxylate:

[0255] To a solution of (Z)-1-(3-((3-(chloromethyl)-1H-isochrom-1-ylidene)amino)-4-isopropoxyphenyl)ethane-1-one 19-7 (150 mg, 0.41 mmol), tert-butyl piperazine-1-carboxylate 19-8 (75 mg, 0.41 mmol) and potassium carbonate (112 mg, 0.81 mmol) in acetonitrile (20 mL) was added potassium iodide (67 mg, 0.41 mmol), and the mixture was stirred at 80° C. for 16 hours. The mixture was concentrated in vacuo, and the residue was diluted with ethyl acetate (50 mL) and washed with saturated brine (30 mL×2). The separated organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (dichloromethane / methanol=20 / 1) to give the yellow solid product 19-9 (100 mg, 45%).

[0256] LC-MS: [M+H] + :520.2.

[0257] 1HNMR: (400MHz, CDCl3) δ8.47(d,J=7.6Hz,1H),7.71(d,J=8.0Hz,1H),7.68-7.64( m,2H),7.57(t,J=7.6Hz,1H),7.38(d,J=7.6Hz,1H),6.99(d,J=8.0Hz,1H),6.68(s ,1H),4.59-4.49(m,1H),4.14-3.98(m,2H),3.81(s,2H),3.61-3.41(m,2H),3.21 -3.10(m,2H),2.84-2.70(m,2H),2.56(s,3H),1.46(s,9H),1.22(d,J=6.0Hz,6H).

[0258] Synthesis of 2-(5-acetyl-2-isopropoxyphenyl)-3-(piperazin-1-ylmethyl)isoquinolin-1(2H)-one:

[0259] To a solution of (Z)-tert-butyl 4-((1-((5-acetyl-2-isopropoxyphenyl)imino)-1H-isochrom-3-yl)methyl)piperazine-1-carboxylate 19-9 (90 mg, 0.077 mmol) in dichloromethane (3 mL) was added trifluoroacetic acid (1 mL), and the mixture was stirred at room temperature for 16 hours. The mixture was concentrated in vacuo to give a mixture of crude yellow solid products 19-10 and 19-11 (70 mg, crude).

[0260] LC-MS: [M+H] + :420.2.

[0261] Synthesis of 2-(5-acetyl-2-isopropoxyphenyl)-3-((4-(2-(4-chlorophenoxy)acetyl)piperazin-1-yl)methyl)isoquinolin-1(2H)-one:

[0262] To a dichloromethane (20 mL) solution of a mixture of 19-10 and 19-11 (60 mg, 0.14 mmol) and N, N-diisopropylethylamine (55.4 mg, 0.43 mmol) was added 2-(4-chlorophenoxy)acetyl chloride 1-10 (29.3 mg, 0.14 mmol), and the mixture was stirred at room temperature for 2 hours. The mixture was diluted with dichloromethane (30 mL) and washed with saturated brine (30 mL). The separated organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by preparative plate chromatography (dichloromethane / methanol = 20 / 1) and efficient preparative liquid chromatography (acetonitrile-water (0.05% ammonia), gradient 30%-90%) to give the product 19-bp (9.9 mg, 11.7%) as a white solid and a white solid product, compound 150 (11.2 mg, 13.2%).

[0263] Example 19-bp

[0264] LC-MS: [M+H] + :588.3.

[0265] HPLC: 99.54% [214 nm]

[0266] 1 H NMR: (400MHz, CD3OD) δ8.33(d,J=8.0Hz,1H),7.79(dd,J=8.4,2.0Hz,1H),7.69-7.63(m,2H),7.55-7.49(m,1H),7.42(d,J=7.6Hz,1H),7.27(d,J =8.8Hz,2H),7.12(d,J=8.8Hz,1H),6.95(d,J=8.8Hz,2H),6.43(s,1H),4.78(s,2H),4.75-4.68(m,1H),3.5 2-3.46(m,4H),3.31-3.30(m,2H),2.58(s,3H),2.48-2.44(m,2H),2.43-2.37(m,2H),1.28(d,J=6.0Hz,6H).

[0267] Example 19 (Compound 150):

[0268] LC-MS: [M+H] + :588.0.

[0269] HPLC: 98.09% [214 nm].

[0270] 1H NMR: (400MHz, CD3OD) δ8.31(d,J=8.0Hz,1H),8.17(dd,J=8.8,2.0Hz,1H),8.04(d,J=2 .0Hz,1H),7.79(t,J=7.6Hz,1H),7.71(d,J=7.6Hz,1H),7.57(t,J=7.6Hz,1H),7.31-7. 25(m,3H),6.93(d,J=8.8Hz,2H),6.82(s,1H),4.83-4.76(m,3H),3.48-3.38(m,4H),3. 19-3.10(m,2H),2.62(s,3H),2.42-2.30(m,2H),2.16-2.05(m,2H),1.26-1.20(m,6H).

[0271] Example 20

[0272] Synthesis of 3-methyl-1H-isochrome-1-one:

[0273] A mixture of 2-iodobenzoic acid 20-1 (2 g, 8.1 mmol), pentane-2,4-dione 20-2 (807 mg, 8.1 mmol), cuprous iodide (150 mg, 0.81 mmol) and potassium phosphate (3.44 g, 16.1 mmol) in N, N-dimethylformamide (20 mL) was heated at 100 ° C for 16 hours. After cooling to room temperature, the mixture was diluted with water (20 mL) and extracted with ethyl acetate (30 mL × 3). The combined organic layer was washed with saturated brine (50 mL × 3). The separated organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether = 1 / 3) to give a brown solid product 20-3 (1 g, 73%).

[0274] LC-MS: [M+H] + :161.2.

[0275] 1 H NMR: (400MHz, DMSO-d6) δ8.10 (d, J = 8.4Hz, 1H), 7.87-7.75 (m, 1H), 7.55-7.52 (m, 2H), 2.25 (s, 3H).

[0276] Synthesis of 2-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)-3-methylisoquinolin-1(2H)-one:

[0277] To a mixture of 4-fluoro-2-isopropoxy-5-(trifluoromethyl)aniline 1-4 (296 mg, 1.25 mmol) in toluene (10 mL) was added trimethylaluminum (1.25 mL, 2.5 mmol, 2 mol in hexane) and stirred at room temperature for 10 minutes. 3-Methylisochrome-1-one 20-3 (200 mg, 1.25 mol) was then added and the mixture was stirred at 80° C. for 16 hours. The mixture was concentrated in vacuo, and the residue was diluted with ethyl acetate (50 mL) and washed sequentially with aqueous sodium bicarbonate solution (20 mL) and saturated brine (30 mL). The separated organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether = 1 / 4) to give the product 20-4 (100 mg, 20%) as a yellow oil.

[0278] LC-MS: [M+H] + :380.1.

[0279] 1 HNMR: (400MHz, CDCl3) δ8.33(d,J=8.0Hz,1H),7.67-7.59(m,1H),7.51-7.40(m,3H),6.85(d,J=12.0Hz,1 H), 6.41 (s, 1H), 4.54 (dt, J = 12.2, 6.0Hz, 1H), 1.98 (s, 3H), 1.26 (d, J = 6.0Hz, 3H), 1.22 (d, J = 6.0Hz, 3H).

[0280] Synthesis of 2-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)-1-oxo-1,2-dihydroisoquinoline-3-carbaldehyde:

[0281] To a solution of 2-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)-3-methylisoquinolin-1(2H)-one 20-4 (100 mg, 0.26 mmol) in dioxane (20 mL) was added selenium dioxide (29 mg, 0.26 mmol), and the mixture was heated at 70°C for 16 hours. After cooling to room temperature, the mixture was concentrated in vacuo, and the residue was diluted with aqueous sodium bicarbonate solution (10 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product 20-5 (100 mg, 77%) as a yellow solid.

[0282] LC-MS: [M+H] + :394.0.

[0283] Synthesis of 2-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)-3-(hydroxymethyl)isoquinolin-1(2H)-one:

[0284] To a solution of 2-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)-1-oxo-1,2-dihydroisoquinoline-3-carbaldehyde 20-5 (100 mg, 0.25 mmol) in methanol (10 mL) was added sodium borohydride (10 mg, 0.25 mmol), and the mixture was stirred at room temperature for 1 hour. The mixture was concentrated in vacuo, the residue was diluted with water (20 mL), and extracted with ethyl acetate (30 mL x 2). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product 20-6 (100 mg, 94%) as a yellow solid.

[0285] LC-MS: [M+H] + :396.1.

[0286] Synthesis of 3-(chloromethyl)-2-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)isoquinolin-1(2H)-one:

[0287] To a solution of 2-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)-3-(hydroxymethyl)isoquinolin-1(2H)-one 20-6 (100 mg, 0.25 mmol) and N,N-diisopropylethylamine (65 mg, 0.50 mmol) in dichloromethane (10 mL) was added methanesulfonyl chloride (19 mg, 0.25 mmol), and the mixture was stirred at room temperature for 2 hours. The mixture was diluted with dichloromethane (30 mL) and washed with saturated brine (30 mL). The separated organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product 20-7 (100 mg, 91%) as a yellow solid.

[0288] LC-MS: [M+H] + :414.0.

[0289] Synthesis of (S)-tert-butyl 4-((2-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)-1-oxo-1,2-dihydroisoquinolin-3-yl)methyl)-3-methylpiperazine-1-carboxylate:

[0290] To a solution of 3-(chloromethyl)-2-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)isoquinolin-1(2H)-one 20-7 (100 mg, 0.24 mmol), (S)-tert-butyl 3-methylpiperazine-1-carboxylate 1-7 (48.7 mg, 0.24 mmol) and potassium carbonate (33 mg, 0.24 mmol) in acetonitrile (30 mL) was added potassium iodide (40 mg, 0.24 mmol), and the mixture was heated at 70° C. for 16 hours. The mixture was concentrated in vacuo, and the residue was diluted with ethyl acetate (50 mL) and washed with saturated brine (30 mL). The separated organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was subjected to silica gel chromatography (ethyl acetate / petroleum ether = 1 / 2) to give the crude product 20-8 (100 mg, 64%) as a yellow solid.

[0291] LC-MS: [M+H] + :578.3.

[0292] Synthesis of (S)-2-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)-3-((2-methylpiperazin-1-yl)methyl)isoquinolin-1(2H)-one:

[0293] To a solution of (S)-tert-butyl 4-((2-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)-1-oxo-1,2-dihydroisoquinolin-3-yl)methyl)-3-methylpiperazine-1-carboxylate 20-8 (100 mg, 0.17 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (1 mL), and the mixture was stirred at room temperature for 16 hours. The mixture was concentrated in vacuo to give the crude product 20-9 (80 mg, 98%) as a yellow oil.

[0294] LC-MS: [M+H] + :478.2.

[0295] Synthesis of (S)-3-((4-(2-(4-chlorophenoxy)acetyl)-2-methylpiperazin-1-yl)methyl)-2-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)isoquinolin-1(2H)-one:

[0296] To a solution of (S)-2-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)-3-((2-methylpiperazin-1-yl)methyl)isoquinolin-1(2H)-one 20-9 (80 mg, 0.17 mmol) and N,N-diisopropylethylamine (65 mg, 0.50 mmol) in dichloromethane (10 mL) was added 2-(4-chlorophenoxy)acetyl chloride (34 mg, 0.17 mmol), and the mixture was stirred at room temperature for 2 hours. The mixture was diluted with dichloromethane (30 mL) and washed with saturated brine (30 mL). The separated organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether = 1 / 1) to give a white solid product, Compound 153 (47 mg, 41%).

[0297] LC-MS: [M+H] + :646.3.

[0298] HPLC: 95.73% [214 nm].

[0299] 1 H NMR: (400MHz, DMSO) δ8.17(d,J=8.0Hz,1H),7.82-7.71(m,2H),7.68(d,J=8.8Hz,1H),7.5 3-7.47(m,1H),7.48-7.40(m,1H),7.31(d,J=8.8Hz,2H),6.91(d,J=8.8Hz,2H),6.74-6.72 (m,1H),4.85-4.69(m,3H),3.64-3.35(m,2H),3.26-3.02(m,1H),2.86-2.75(m,2H),2.62 -2.56(m,1H),2.44-2.34(m,1H),2.15-1.81(m,2H),1.19-1.10(m,6H),0.63-0.42(m,3H).

[0300] Example 21 / 22

[0301] Compound 153 (47 mg) was chiral separated by SFC (chromatographic column: WHELK-O1 250 mm × 21.1 mm, 5 μm, mobile phase: carbon dioxide-ethanol (NH4OH 0.2%) gradient: 0-40%) to obtain the title compound (13.9 mg, 19.4 mg)

[0302] Single configuration compound (shorter retention time)

[0303] It was analyzed by chiral analysis chromatography conditions, RT = 10.35min

[0304] LC-MS: [M+H] + :646.3.

[0305] HPLC: 95.73% [214 nm].

[0306] 1 H NMR: (400MHz, DMSO) δ8.17(d,J=8.0Hz,1H),7.82-7.71(m,2H),7.68(d,J=8.8Hz,1H),7.5 3-7.47(m,1H),7.48-7.40(m,1H),7.31(d,J=8.8Hz,2H),6.91(d,J=8.8Hz,2H),6.74-6.72 (m,1H),4.85-4.69(m,3H),3.64-3.35(m,2H),3.26-3.02(m,1H),2.86-2.75(m,2H),2.62 -2.56(m,1H),2.44-2.34(m,1H),2.15-1.81(m,2H),1.19-1.10(m,6H),0.63-0.42(m,3H).

[0307] Single configuration compound (longer retention time)

[0308] It was analyzed by chiral analysis chromatography conditions, RT = 12.72min

[0309] LC-MS: [M+H] + :646.3.

[0310] HPLC: 95.73% [214 nm].

[0311] 1 H NMR: (400MHz, DMSO) δ8.17(d,J=8.0Hz,1H),7.82-7.71(m,2H),7.68(d,J=8.8Hz,1H),7.5 3-7.47(m,1H),7.48-7.40(m,1H),7.31(d,J=8.8Hz,2H),6.91(d,J=8.8Hz,2H),6.74-6.72 (m,1H),4.85-4.69(m,3H),3.64-3.35(m,2H),3.26-3.02(m,1H),2.86-2.75(m,2H),2.62 -2.56(m,1H),2.44-2.34(m,1H),2.15-1.81(m,2H),1.19-1.10(m,6H),0.63-0.42(m,3H).

[0312] Example 23 / 24

[0313] Compound 78 (13 mg) was chiral separated by SFC (chromatographic column: WHELK-O1 250 mm × 21.1 mm, 5 μm, mobile phase: carbon dioxide-ethanol (NH4OH 0.2%) gradient: 0-40%) to obtain the title compound (2.6 mg, 3.3 mg)

[0314] Single configuration compound (shorter retention time)

[0315] It was analyzed by chiral analysis chromatography conditions, RT = 1.98min

[0316] LC-MS: [M+H] + :657.3.

[0317] HPLC: 98.85% [214 nm].

[0318] 1 H NMR: (400MHz, CD3OD) δ8.20(d,J=8.0Hz,1H),7.92-7.85(m,2H),7.78(d,J=8.0Hz,1H),7.58(t,J=7.6Hz,1H),7.22-7.20(m,3H),6.91 (d,J=8.8Hz,2H),4.77-4.69(m,3H),3.62-3.52(m,4H),3.42-3.32(m,1H),3.28-3.24(m,2H),2.65-2.41(m,4H),1.21-1.09(m,12H).

[0319] Single configuration compound (longer retention time)

[0320] It was analyzed by chiral analysis chromatography conditions, RT = 8.68min

[0321] LC-MS: [M+H] + :657.3.

[0322] HPLC: 98.85% [214 nm].

[0323] 1H NMR: (400MHz, CD3OD) δ8.20(d,J=8.0Hz,1H),7.92-7.85(m,2H),7.78(d,J=8.0Hz,1H),7.58(t,J=7.6Hz,1H),7.22-7.20(m,3H),6.91 (d,J=8.8Hz,2H),4.77-4.69(m,3H),3.62-3.52(m,4H),3.42-3.32(m,1H),3.28-3.24(m,2H),2.65-2.41(m,4H),1.21-1.09(m,12H).

[0324] Example 25

[0325] According to Example 1, 25-1 was substituted for 1-4 to give the current product (S)-5-(2-((4-(2-(4-chlorophenoxy)acetyl)-2-methylpiperazin-1-yl)methyl)-4-oxoquinazolin-3(4H)-yl)-4-isopropoxy-2-methylbenzoic acid methyl ester (25.3 mg, 64.6% yield)

[0326] LC-MS: [M+H]+: 633.0.

[0327] HPLC: 100%.

[0328] 1 H NMR: (400MHz, CD3OD) δ8.24(d,J=8.0Hz,1H),8.02-7.99(m,1H),7.93-7.88(m,1H),7.77(d,J=8.3H z,1H),7.60(t,J=7.6Hz,1H),7.28(d,J=8.6Hz,2H),7.15(s,1H),6.97-6.92(m,2H),4.83-4.72(m, 3H),3.86(d,J=4.6Hz,3H),3.82-3.61(m,2H),3.52-3.46(m,1H),3.27-2.98(m,2H),2.89-2.61(m, 5H),2.45-2.18(m,2H),1.27(dd,J=6.0,3.1Hz,3H),1.19(dd,J=6.0,1.8Hz,3H),0.81-0.58(m,3H).

[0329] Example 26

[0330] According to Example 1, 26-2 was replaced with 1-4 to give the current product (S)-3-(2-((4-(2-(4-chlorophenoxy)acetyl)-2-methylpiperazin-1-yl)methyl)-4-oxoquinazolin-3(4H)-yl)-4-isopropoxy-2-methylbenzoic acid methyl ester (21.0 mg, 41.7% yield)

[0331] LC-MS: [M+H]+: 633.0.

[0332] HPLC:94.76%.

[0333] 1H NMR: (400MHz, CD3OD) δ8.26(d,J=8.0Hz,1H),8.11(d,J=7.6Hz,1H),7.92(t,J=7.6Hz,1H),7.81 -7.79(m,1H),7.62(t,J=7.6Hz,1H),7.27(d,J=8.8Hz,2H),7.17(t,J=8.0Hz,1H),6.95-6.93(m ,2H),4.80-4.71(m,3H),3.91(s,3H),3.73-3.41(m,3H),3.26-3.15(m,1H),3.07-2.92(m,1H), 2.81-2.04(m,7H),1.29-1.24(m,3H),1.16-1.12(m,3H),0.83-0.78(m,2H),0.49-0.44(m,1H).

[0334] Example 27

[0335] Referring to the steps of Example 1, 27-1 was substituted for 1-7 to obtain the target compound (R)-2-((4-(2-(4-chlorophenoxy)acetyl)-2-methylpiperazin-1-yl)methyl)-3-(4-fluoro-2-isopropoxy-5-(trifluoromethyl)phenyl)quinazolin-4(3H)-one (60.7 mg, yield: 31.0%, purity: 97.01%).

[0336] LC-MS: [M+H] + :647.3.

[0337] HPLC: 97.01%

[0338] 1H NMR(400MHz,CD3OD)δ8.20(d,J=7.2Hz,1H),7.91-7.80(m,2H),7.71(dd,J=10.2 ,5.8Hz,1H),7.60-7.54(m,1H),7.34-7.19(m,3H),6.95-6.82(m,2H),4.80-4.61 (m,3H),3.93-3.69(m,1H),3.65-3.36(m,2H),3.25-3.07(m,2H),3.02-2.53(m, 2H),2.39-2.06(m,2H),1.29-1.19(m,3H),1.18-1.16(m,3H),0.63-0.59(m,3H).

[0339] Example 28 / 29

[0340] Compound 140 (60.7 mg) was chiral separated by SFC (chromatographic column: AD-H 250 mm × 20 mm, 5 μm, mobile phase: carbon dioxide-ethanol (NH4OH 0.2%) gradient: 0-40%) to obtain the title compound (18.9 mg, 17.6 mg)

[0341] Single configuration compound (shorter retention time)

[0342] It was analyzed by chiral analysis chromatography conditions, RT = 2.04min

[0343] LC-MS: [M+H]+: 647.3.

[0344] HPLC: 97.01%

[0345] 1 H NMR(400MHz,CD3OD)δ8.20(d,J=7.2Hz,1H),7.91-7.80(m,2H),7.71(dd,J=10.2 ,5.8Hz,1H),7.60-7.54(m,1H),7.34-7.19(m,3H),6.95-6.82(m,2H),4.80-4.61 (m,3H),3.93-3.69(m,1H),3.65-3.36(m,2H),3.25-3.07(m,2H),3.02-2.53(m, 2H),2.39-2.06(m,2H),1.29-1.19(m,3H),1.18-1.16(m,3H),0.63-0.59(m,3H).

[0346] Single configuration compound (longer retention time)

[0347] It was analyzed by chiral analysis chromatography conditions, RT = 3.13min

[0348] LC-MS: [M+H]+: 647.3.

[0349] HPLC: 97.60%

[0350] 1 H NMR(400MHz,CD3OD)δ8.20(d,J=7.2Hz,1H),7.91-7.80(m,2H),7.71(dd,J=10.2 ,5.8Hz,1H),7.60-7.54(m,1H),7.34-7.19(m,3H),6.95-6.82(m,2H),4.80-4.61 (m,3H),3.93-3.69(m,1H),3.65-3.36(m,2H),3.25-3.07(m,2H),3.02-2.53(m, 2H),2.39-2.06(m,2H),1.29-1.19(m,3H),1.18-1.16(m,3H),0.63-0.59(m,3H).

[0351] Example 30

[0352] Synthesis of 4-(methylthio)-2-nitrophenol:

[0353] To a solution of 4-(methylthio)phenol (500mg, 3.57mmol) in acetonitrile (20mL) was added tert-butyl nitrite (331mg, 3.21mmol), and the mixture was stirred at room temperature for 4 hours. The mixture was concentrated under reduced pressure, and the residue was diluted with ethyl acetate (30mL) and washed with saturated brine (30mL). The separated organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether=1 / 4) to give the product 30-2 (200mg, yield: 29%) as a yellow solid.

[0354] LC-MS: [M+H] + :No MS.

[0355] 1 H NMR: (400MHz, CDCl3) δ10.50 (s, 1H), 7.96 (d, J = 2.4Hz, 1H), 7.52 (dd, J = 8.8, 2.4Hz, 1H), 7.11 (d, J = 8.8Hz, 1H), 2.51 (s, 3H).

[0356] Synthesis of 1-isopropoxy-4-(methylthio)-2-nitrobenzene:

[0357] To a solution of 4-(methylthio)-2-nitrophenol (100 mg, 0.54 mmol) and potassium carbonate (149 mg, 1.08 mmol) in N, N-dimethylformamide (10 mL) was added 2-iodopropane (137.7 mg, 0.81 mmol), and the mixture was heated at 60 ° C for 16 hours. After cooling to room temperature, the mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layer was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether=1 / 3) to give a yellow oily product 30-3 (100 mg, yield: 77.4%).

[0358] LC-MS: [M+H] + :No MS.

[0359] 1 HNMR: (400MHz, CDCl3) δ7.66(d,J=2.4Hz,1H),7.40(dd,J=8.8,2.4Hz,1H),7.00(d,J=8.8Hz,1H),4.61(dt,J=12.4,6.0Hz,1H),1.37(d,J=6.0Hz,6H).

[0360] Synthesis of 2-isopropoxy-5-(methylthio)aniline:

[0361] A mixture of 1-isopropoxy-4-(methylthio)-2-nitrobenzene (90 mg, 0.40 mmol) and palladium on carbon (11 mg, 20% wet) in ethanol (10 mL) was hydrogenated under a hydrogen atmosphere for 2 hours. The mixture was filtered and the filtrate was concentrated under reduced pressure to give the yellow oily product 30-4 (75 mg, yield: 86%).

[0362] LC-MS: [M+H] + :198.1.

[0363] Synthesis of 2-(chloromethyl)-3-(2-isopropoxy-5-(methylthio)phenyl)quinazolin-4(3H)-one:

[0364] To a solution of 2-(2-chloroacetylamino)benzoic acid (80 mg, 0.37 mmol) in acetonitrile (20 mL) was added N, N-diisopropylethylamine (48 mg, 0.37 mmol) and phosphorus trichloride (77 mg, 0.56 mmol), and the mixture was stirred at room temperature for 10 minutes. A solution of 2-isopropoxy-5-(methylthio)aniline (74 mg, 0.37 mmol) in acetonitrile (1 mL) was added, and the mixture was stirred at 60 ° C for 2 hours. After cooling to room temperature, the mixture was concentrated in vacuo, the residue was diluted with sodium bicarbonate aqueous solution (10 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layer was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether=1 / 3) to give product 30-5 (70 mg, yield: 90%) as a yellow solid.

[0365] LC-MS: [M+H] + :375.1.

[0366] Synthesis of tert-butyl 4-((3-(2-isopropoxy-5-(methylthio)phenyl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylate:

[0367] To a solution of 2-(chloromethyl)-3-(2-isopropoxy-5-(methylthio)phenyl)quinazolin-4(3H)-one (70 mg, 0.19 mmol), tert-butyl piperazine-1-carboxylate (35 mg, 0.19 mmol) and potassium carbonate (52 mg, 0.37 mmol) in acetonitrile (20 mL) was added potassium iodide (31 mg, 0.19 mmol), and the mixture was stirred at 80° C. for 16 hours. The mixture was concentrated in vacuo, and the residue was diluted with ethyl acetate (30 mL) and washed with saturated brine (20 mL). The separated organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (dichloromethane / methanol=20 / 1) to give the product 30-6 (50 mg, yield: 87%) as a yellow solid.

[0368] LC-MS: [M+H] + :525.2.

[0369] Synthesis of tert-butyl 4-((3-(2-isopropoxy-5-(S-methylsulfonylamino)phenyl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylate:

[0370] To a solution of tert-butyl 4-((3-(2-isopropoxy-5-(methylthio)phenyl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylate (80 mg, 0.15 mmol) in methanol (20 mL) was added ammonium carbamate (47.6 mg, 0.61 mmol) and iodophenyl diacetate (147 mg, 0.45 mmol), and the mixture was stirred at room temperature for 16 hours. The mixture was concentrated in vacuo, the residue was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (dichloromethane / methanol=15 / 1) to give product 30-7 (40 mg, yield: 42.5%) as a yellow solid.

[0371] LC-MS: [M+H] + :556.2.

[0372] Synthesis of 3-(2-isopropoxy-5-(S-methylsulfonylanilide)phenyl)-2-(piperazin-1-ylmethyl)quinazolin-4(3H)-one:

[0373] To a solution of tert-butyl 4-((3-(2-isopropoxy-5-(S-methylsulfonylamino)phenyl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylate (40 mg, 0.072 mmol) in dichloromethane (3 mL) was added trifluoroacetic acid (1 mL), and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated in vacuo to give the crude product 30-8 (30 mg, yield: 82%) as a yellow solid.

[0374] LC-MS: [M+H] + :456.2.

[0375] Synthesis of 2-((4-(2-(4-chlorophenoxy)acetyl)piperazin-1-yl)methyl)-3-(2-isopropoxy-5-(S-methylsulfonylimino)phenyl)quinazolin-4(3H)one:

[0376] To a solution of 3-(2-isopropoxy-5-(S-methylsulfonylanilide)phenyl)-2-(piperazin-1-ylmethyl)quinazolin-4(3H)-one (30 mg, 0.066 mmol) and N,N-diisopropylethylamine (26 mg, 0.20 mmol) in dichloromethane (10 mL) was added 2-(4-chlorophenoxy)acetyl chloride (13.5 mg, 0.066 mmol), and the mixture was stirred at room temperature for 2 hours. The mixture was diluted with dichloromethane (30 mL) and washed with saturated brine (20 mL). The separated organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by preparative plate (dichloromethane / methanol = 15 / 1) to give the product compound 189 (34 mg, yield: 80%) as a white solid.

[0377] LC-MS: [M+H] + :624.3.

[0378] HPLC: 97.13%.

[0379] 1 H NMR: (400MHz, DMSO-d6)δ8.15(d,J=8.0Hz,1H),8.05-7.97(m,2H),7.88(t,J=7.6 Hz,1H),7.74(d,J=8.0Hz,1H),7.58(t,J=7.6Hz,1H),7.42(d,J=8.8Hz,1H),7.31 (d,J=8.8Hz,2H),6.90(d,J=8.8Hz,2H),4.85-4.76(m,3H),4.27(s,1H),3.46-3. 38(m,1H),3.29-3.14(m,5H),3.12(s,3H),2.25-2.02(m,4H),1.17-1.11(m,6H).

[0380] Example 31

[0381] Synthesis of 3-(2-isopropoxy-5-(methylthio)phenyl)-2-(piperazin-1-ylmethyl)quinazolin-4(3H)-one:

[0382] A mixture of 2-(chloromethyl)-3-(2-isopropoxy-5-(methylthio)phenyl)quinazolin-4(3H)-one (50 mg, 0.14 mmol) and piperazine (34.5 mg, 0.40 mmol) in THF (20 mL) was heated at reflux for 16 hours. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with aqueous sodium bicarbonate (10 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 31-1 (60 mg) as a white solid, which was used directly in the next step.

[0383] LC-MS: [M+H] + :425.1.

[0384] Synthesis of 2-((4-(2-(4-chlorophenoxy)acetyl)piperazin-1-yl)methyl)-3-(2-isopropoxy-5-(methylthio)phenyl)quinazolin-4(3H)one:

[0385] To a solution of 3-(2-isopropoxy-5-(methylthio)phenyl)-2-(piperazin-1-ylmethyl)quinazolin-4(3H)-one (60 mg, 0.14 mmol) in dichloromethane (20 mL) were added N,N-diisopropylethylamine (27.4 mg, 0.21 mmol) and 2-(4-chlorophenoxy)acetyl chloride (29.0 mg, 0.14 mmol), and the mixture was stirred at room temperature for 6 hours. The residue was extracted with aqueous sodium bicarbonate (20 mL) and dichloromethane (30 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by high-performance preparative liquid chromatography to afford compound 180 (61.6 mg, 71% yield) as a white solid.

[0386] LC-MS: [M+H] + :593.0.

[0387] HPLC:95.67%.

[0388] 1H NMR: (400MHz, DMSO-d6)δ8.14(dd,J=8.0,1.2Hz,1H),7.89-7.85(m,1H),7.72(d,J=8.0Hz,1H ),7.59-7.55(m,1H),7.44(d,J=2.4Hz,1H),7.38(dd,J=8.8,2.4Hz,1H),7.33-7.28(m,2H),7. 20(d,J=8.8Hz,1H),6.93-6.88(m,2H),4.78(s,2H),4.63-4.56(m,1H),3.30-3.15(m,6H),2. 47(s,3H),2.38-2.24(m,2H),2.11-2.01(m,2H),1.13(d,J=6.0Hz,3H),1.05(d,J=6.0Hz,3H).

[0389] Example 32

[0390] Synthesis of 1-isopropoxy-4-(methylsulfonyl)-2-nitrobenzene:

[0391] To a solution of 4-(methylsulfonyl)-2-nitrophenol (200.0 mg, 0.92 mmol) in DMF (10 mL) was added potassium carbonate (152.8 mg, 1.11 mmol). After stirring for 15 minutes, 2-iodopropane (313.1 mg, 1.85 mmol) was added, and the mixture was stirred at 70°C for 16 hours. The mixture was diluted with water (30 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed three times with saturated sodium chloride solution (60 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether = 1 / 1) to afford product 32-2 (170 mg, yield: 68%) as a white solid.

[0392] LC-MS: [M+Na] + :282.0.

[0393] 1 H NMR: (400MHz, DMSO-d6)δ8.38(d,J=2.3Hz,1H),8.12(dd,J=8.9,2.4Hz,1H), 7.65(d,J=9.0Hz,1H),5.02-4.96(m,1H),3.28(s,3H),1.33(d,J=6.0Hz,6H).

[0394] Synthesis of 2-isopropoxy-5-(methylsulfonyl)aniline:

[0395] To a solution of 1-isopropoxy-4-(methylsulfonyl)-2-nitrobenzene (150 mg, 0.58 mmol) in methanol (20 mL) was added palladium hydroxide on carbon (467.0 mg, 20% water content, 0.38 mmol). The mixture was reacted under hydrogen (0.4 MPa) for 3 hours. The filtrate was then collected by filtration and dried under vacuum to afford the product 32-2 (110.0 mg, 75% yield) as a white solid.

[0396] LC-MS: [M+H] + :230.1.

[0397] Synthesis of 2-(chloromethyl)-3-(2-isopropoxy-5-(methylsulfonyl)phenyl)quinazolin-4(3H)-one:

[0398] To a solution of 2-(2-chloroacetylamino)benzoic acid (94.0 mg, 0.44 mmol) in acetonitrile (20 mL) was added N,N-diisopropylethylamine (68.3 mg, 0.53 mmol). After stirring for 2 minutes, phosphorus trichloride (151.1 mg, 1.10 mmol) was added. After stirring for 10 minutes, a solution of 2-isopropoxy-5-(methylsulfonyl)aniline (100.9 mg, 0.44 mmol) in acetonitrile (2 mL) was added. The mixture was heated at reflux for 6 hours. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with aqueous sodium bicarbonate (20 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel chromatography (dichloromethane / methanol = 15 / 1) to afford product 32-3 (140 mg, yield: 72%) as a white solid.

[0399] LC-MS: [M+H] + :407.0.

[0400] Synthesis of 3-(2-isopropoxy-5-(methylsulfonyl)phenyl)-2-(piperazin-1-ylmethyl)quinazolin-4(3H)-one:

[0401] 2-(Chloromethyl)-3-(2-isopropoxy-5-(methylsulfonyl)phenyl)quinazolin-4(3H)-one (140 mg, 0.34 mmol) and piperazine (88.9 mg, 1.03 mmol) were dissolved in THF (20 mL), and the mixture was heated at reflux for 16 hours. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with aqueous sodium bicarbonate (10 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to afford 32-8 (130 mg crude) as a white solid, which was used directly in the next reaction.

[0402] LC-MS: [M+H] + :457.2.

[0403] Synthesis of 2-((4-(2-(4-chlorophenoxy)acetyl)piperazin-1-yl)methyl)-3-(2-isopropoxy-5-(methylsulfonyl)phenyl)quinazolin-4(3H)one:

[0404] 3-(2-isopropoxy-5-(methylsulfonyl)phenyl)-2-(piperazin-1-ylmethyl)quinazolin-4(3H)-one

[0405] To a solution of 140 mg, 0.30 mmol, N,N-diisopropylethylamine (59.4 mg, 0.46 mmol) and 2-(4-chlorophenoxy)acetyl chloride (62.9 mg, 0.30 mmol) in dichloromethane (20 mL) were added, and the mixture was stirred at room temperature for 6 hours. The residue was extracted with aqueous sodium bicarbonate (20 mL) and dichloromethane (30 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by high-performance preparative liquid chromatography to afford compound 183 (125.2 mg, yield: 66%) as a white solid.

[0406] LC-MS: [M+H]+: 625.2.

[0407] HPLC: 100%.

[0408] 1H NMR: (400MHz, DMSO-d6) δ8.15(d,J=8.0Hz,1H),8.10(d,J=2.4Hz,1H),8.00(dd,J=8.8,2.4Hz,1H),7.89-7.86(m 1H),7.74(d,J=8.0Hz,1H),7.59(t,J=7.6Hz,1H),7.47(d,J=9.0Hz,1H),7.32-7.29(m,2H),6.92-6.88(m,2H),4.84-4.80(m,1H),4. 77(s,2H),3.41-3.30(m,5H),3.24-3.13(m,4H),2.29-2.23(m,1H),2.17-2.05(m,3H),1.17(d,J=6.0Hz,3H),1.12(d,J=6.0Hz,3H).

[0409] Example 33 / 34

[0410] Compound 183 (125 mg) was chiral separated by SFC (chromatographic column: WHELK-O1 250 mm × 21.1 mm, 5 μm, mobile phase: carbon dioxide-ethanol (NH4OH 0.2%) gradient: 0-40%) to obtain the title compound (47 mg, 48.8 mg)

[0411] Single configuration compound (shorter retention time)

[0412] It was analyzed by chiral analysis chromatography conditions, RT = 3.29min

[0413] LC-MS: [M+H]+: 625.2.

[0414] HPLC:99.72%.

[0415] 1H NMR: (400MHz, DMSO-d6) δ8.15(d,J=8.0Hz,1H),8.10(d,J=2.4Hz,1H),8.00(dd,J=8.8,2.4Hz,1H),7.89-7.86(m 1H),7.74(d,J=8.0Hz,1H),7.59(t,J=7.6Hz,1H),7.47(d,J=9.0Hz,1H),7.32-7.29(m,2H),6.92-6.88(m,2H),4.84-4.80(m,1H),4. 77(s,2H),3.41-3.30(m,5H),3.24-3.13(m,4H),2.29-2.23(m,1H),2.17-2.05(m,3H),1.17(d,J=6.0Hz,3H),1.12(d,J=6.0Hz,3H).

[0416] Single configuration compound (longer retention time)

[0417] It was analyzed by chiral analysis chromatography conditions, RT = 4.29min

[0418] LC-MS: [M+H]+: 625.2.

[0419] HPLC: 100%.

[0420] 1 H NMR: (400MHz, DMSO-d6) δ8.15(d,J=8.0Hz,1H),8.10(d,J=2.4Hz,1H),8.00(dd,J=8.8,2.4Hz,1H),7.89-7.86(m 1H),7.74(d,J=8.0Hz,1H),7.59(t,J=7.6Hz,1H),7.47(d,J=9.0Hz,1H),7.32-7.29(m,2H),6.92-6.88(m,2H),4.84-4.80(m,1H),4. 77(s,2H),3.41-3.30(m,5H),3.24-3.13(m,4H),2.29-2.23(m,1H),2.17-2.05(m,3H),1.17(d,J=6.0Hz,3H),1.12(d,J=6.0Hz,3H).

[0421] Example 35

[0422] Referring to Example 1, 35-3 was substituted for 1-4 to obtain the target compound 186 (13.48 mg)

[0423] LC-MS: [M+H]+: 626.0.

[0424] HPLC:99.66%.

[0425] 1 HNMR (400MHz, CD3OD) δ8.28(d,J=8.0Hz,1H),8.10(dd,J=8.8,2.2Hz,1H),8.00(d,J=2.2Hz,1H),7.96(t,J=7.6Hz ,1H),7.87(d,J=8.0Hz,1H),7.66(t,J=7.6Hz,1H),7.45(d,J=8.8Hz,1H),7.28(d,J=8.8Hz,2H),6.96(d,J=9.0Hz, 2H),4.87-4.80(m,3H),4.00-3.96(m,1H),3.88-3.67(m,5H),3.20-2.90(m,4H),1.29(d,J=6.0Hz,3H),1.20(d,J=6.0Hz,3H).

[0426] Example 36

[0427] Synthesis of 4-bromo-1-isopropoxy-2-nitrobenzene:

[0428] To a solution of 4-bromo-2-nitrophenol (2.5 g, 11.5 mmol) in N,N-dimethylformamide (50 mL) at room temperature were added 2-iodoisopropyl (3.91 g, 23 mmol) and potassium carbonate (3.18 g, 23 mmol). The mixture was stirred at room temperature for 24 hours. The reaction solution was diluted with water (500 mL) and extracted with ethyl acetate (100 mL x 3). The combined organic layers were washed with saturated brine (250 mL x 2). The separated organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 5:1) to obtain the yellow oily product 36-2 (2.7 g, yield: 91.0%).

[0429] 1 H NMR (400MHz, DMSO-d6) δ8.08(d,J=2.5Hz,1H),7.79(dd,J=9.0,2.5Hz,1H),7.38(d,J=9.1Hz,1H),4.82(dt,J=12.1,6.0Hz,1H),1.28(d,J=6.0Hz,6H).

[0430] Synthesis of 5-bromo-2-isopropoxyaniline:

[0431] To a solution of 4-bromo-1-isopropoxy-2-nitrobenzene (500 mg, 1.92 mmol) in ethanol / saturated ammonium chloride (20 / 10 mL) was added reduced iron powder (215 mg, 3.84 mmol) at room temperature, and the mixture was stirred at 70°C for 16 hours. The reaction mixture was filtered through celite, and the filtrate was concentrated under reduced pressure. The concentrate was diluted with water (50 mL), and the pH was adjusted to 9-10 by adding saturated sodium bicarbonate solution. The mixture was extracted with dichloromethane (100 mL x 2), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford crude product 36-3 (450 mg) as a yellow oil, which was used directly in the next reaction.

[0432] LC-MS: [M+H] + :230.1.

[0433] Synthesis of 3-(5-bromo-2-isopropoxyphenyl)-2-chloromethylquinazol-4(3H)one:

[0434] To a solution of 2-(2-chloroacetylamino)benzoic acid (140 mg, 0.66 mmol) in dioxane (10 mL) at 0°C were added N,N-diisopropylethylamine (101 mg, 0.79 mmol) and phosphorus trichloride (135 mg, 0.98 mmol). The mixture was stirred at room temperature for 20 minutes, followed by the addition of 5-bromo-2-isopropoxyaniline (151 mg, 0.66 mmol). The mixture was heated to 70°C and stirred for 6 hours. The reaction mixture was concentrated under reduced pressure and diluted with dichloromethane (50 mL). The organic phase was washed with water (50 mL x 2) and saturated brine (100 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (dichloromethane:methanol = 20:1) to afford 36-4 as a yellow solid (260 mg, yield: 97%).

[0435] LC-MS: [M+H]+: 407.5.

[0436] Synthesis of tert-butyl 4-(3-(5-bromo-2-isopropoxyphenyl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylate:

[0437] To a solution of 3-(5-bromo-2-isopropoxyphenyl)-2-chloromethylquinazol-4(3H)one (270 mg, 0.66 mmol) in acetonitrile (10 mL) were added tert-butyl piperazine-1-carboxylate (124 mg, 0.66 mmol), potassium iodide (132 mg, 0.8 mmol), and potassium carbonate (119 mg, 0.86 mmol) at room temperature. The mixture was stirred at 80°C for 16 hours. The reaction solution was concentrated under reduced pressure and diluted with dichloromethane (100 mL). The mixture was washed with water (50 mL x 3) and saturated brine (50 mL x 2). The separated organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 5:1) to afford 36-5 (300 mg, yield: 81%) as a dark yellow oil.

[0438] LC-MS: [M+H] + :557.1.

[0439] Synthesis of tert-butyl 4-((3-(5-(diphenylmethylene)amino)-2-isopropoxyphenyl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylate:

[0440] To a solution of tert-butyl 4-(3-(5-bromo-2-isopropoxyphenyl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-carboxylate (500 mg, 0.90 mmol) in dioxane (20 mL) was added diphenylketone imine (243 mg, 1.34 mmol), tris(dibenzylacetone)dipalladium (82 mg, 0.09 mmol), 4,5-bis(diphenylphosphino-9,9-dimethylxanthene) (52 mg, 0.09 mmol), and cesium carbonate (583 mg, 1.79 mmol) at room temperature. The mixture was stirred at 90°C for 20 hours. The reaction mixture was concentrated under reduced pressure and diluted with ethyl acetate (100 mL), washed sequentially with water (100 mL x 3) and saturated brine (200 mL). The separated organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 5:1) to give a yellow solid product 36-7 (350 mg, yield: 59.0%).

[0441] LC-MS: [M+H] + :658.3.

[0442] Synthesis of tert-butyl 4-(3-(5-amino-2-isopropoxyphenyl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylate:

[0443] 0.5N aqueous hydrochloric acid (2 mL) was added to a solution of tert-butyl 4-((3-(5-(diphenylmethylene)amino)-2-isopropoxyphenyl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylate (460 mg, 0.70 mmol) in tetrahydrofuran (10 mL). The reaction was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure to afford a yellow oil 36-8 (450 mg crude product), which was used directly in the next reaction.

[0444] LC-MS: [M+H] + :494.1.

[0445] Synthesis of tert-butyl 4-(3-(2-isopropoxy-5-(2,2,2-trifluoroacetylamino)phenyl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylate:

[0446] To a solution of tert-butyl 4-(3-(5-amino-2-isopropoxyphenyl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylate (100 mg, 0.2 mmol) in dichloromethane (10 mL) at 0°C was added N,N-diisopropylethylamine (52 mg, 0.4 mmol) and trifluoroacetic anhydride (55 mg, 0.3 mmol). The mixture was stirred at room temperature for 4 hours. The reaction solution was diluted with dichloromethane (50 mL) and washed sequentially with water (100 mL x 2) and saturated brine (100 mL). The separated organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 10:1) to obtain 36-9 as a yellow oil (115 mg, yield: 96%).

[0447] LC-MS: [M+H] + :590.4.

[0448] Synthesis of 2,2,2-trifluoro-N-(4-isopropoxy-3-(4-oxo-2-(piperazin-1-ylmethyl)quinazoline-3(4H)-phenyl)acetamide:

[0449] Trifluoroacetic acid (2 mL) was added to a solution of tert-butyl 4-(3-(2-isopropoxy-5-(2,2,2-trifluoroacetylamino)phenyl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)piperazine-1-carboxylate (110 mg, 1.86 mmol) in dichloromethane (5 mL). The mixture was stirred at room temperature for 2 hours. The solution was concentrated under reduced pressure to afford 36-10 as a yellow oil (172 mg crude product), which was used directly in the next reaction.

[0450] LC-MS: [M+H] + :490.2.

[0451] Synthesis of N-(3-(2-(4-chlorophenoxy)acetyl)piperazin-1-yl)methyl)-4-oxoquinazolin-3(4H)-yl)-4-isopropoxyphenyl)-2,2,2-trifluoroethylamine:

[0452] To a solution of 2,2,2-trifluoro-N-(4-isopropoxy-3-(4-oxo-2-(piperazin-1-ylmethyl)quinazoline-3(4H)-phenyl)acetamide (95 mg, 0.19 mmol) in dichloromethane (10 mL) were added N,N-diisopropylethylamine (50 mg, 0.39 mmol) and 2-(4-chlorophenoxy)acetyl chloride (52 mg, 0.25 mmol) at 0°C, and the mixture was stirred at room temperature for 4 hours. The reaction solution was diluted with dichloromethane (50 mL) and washed with water (100 mL x 2) and saturated brine (100 mL). The separated organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by HPLC (acetonitrile-water (0.1% formic acid) gradient: 45-50) to obtain compound 192 (18 mg, yield: 14%) as a white solid.

[0453] LC-MS: [M+H] + :658.2.

[0454] HPLC: 98.2%

[0455] 1 HNMR (400MHz, CD3OD) δ8.25(dd,J=8.0,1.2Hz,1H),7.95-7.89(m,1H),7.86-7.81(m,2H),7.70(dd,J=9.0,2.6Hz,1H),7.65-7.60(m,1H),7.32(d, J=9.0Hz,1H),7.26-7.21(m,2H),6.95-6.90(m,2H),4.81(s,2H),4.72-4.65(m,1H),4.26-3. 69(m,5H),3.47-3.29(m,3H),3.26-2.98(m,2H),1.23(d,J=6.0Hz,3H),1.11(d,J=6.0Hz,3H).

[0456] Example 37

[0457] Referring to Example 36, methanesulfonyl chloride was substituted for trifluoroacetic anhydride to obtain the target product Compound 195 (26.3 mg)

[0458] LC-MS: [M+H] + :640.3.

[0459] HPLC: 99.5%

[0460] 1 HNMR (400MHz, CD3OD) δ8.25 (d, J=7.6Hz, 1H), 7.92-7.85 (m, 2H), 7.63 (t, J=7. 6Hz,1H),7.44-7.35(m,2H),7.29-7.23(m,3H),7.00-6.89(m,2H),4.83(s,2H ),4.65-4.62(m,1H),4.35-4.20(m,1H),4.00-3.79(m,4H),3.52-3.21(m,4H) ,3.22-3.19(m,1H),3.00(s,3H),1.22(d,J=6.0Hz,3H),1.09(d,J=6.0Hz,3H).

[0461] Example 38

[0462] Synthesis of 6-hydroxy-7-nitro-3,4-dihydroisoquinolin-1(2H)-one:

[0463] To a solution of 6-hydroxy-3,4-dihydroisoquinolin-1(2H)-one (327.0 mg, 2.00 mmol) in concentrated sulfuric acid (5 mL) at -10°C was added sodium nitrate (170.4 mg, 2.00 mmol) and stirred at -10°C for 2 hours. The reaction was quenched by adding ice, diluted with water (30 mL), and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with saturated sodium chloride solution (60 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether = 1 / 1) to afford product 38-2 (150 mg, yield: 35%) as a white solid.

[0464] LC-MS: [M+H] + :209.0.

[0465] 1 H NMR: (400MHz, DMSO-d6) δ11.58(s,1H),8.29(s,1H),8.03(s,1H),7.01(s,1H),3.42-3.37(m,2H),2.91(d,J=6.4Hz,2H).

[0466] Synthesis of 6-isopropoxy-7-nitro-3,4-dihydroisoquinolin-1(2H)-one:

[0467] To a solution of 6-hydroxy-7-nitro-3,4-dihydroisoquinolin-1(2H)-one (150.0 mg, 0.72 mmol) in N,N-dimethylformamide (10 mL) was added potassium carbonate (199.2 mg, 1.44 mmol). After stirring for 15 minutes, 2-iodopropane (183.8 mg, 1.08 mmol) was added, and the mixture was stirred at 70°C for 16 hours. The mixture was diluted with water (30 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with saturated sodium chloride solution (60 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether = 1 / 1) to afford product 38-3 (170 mg, yield: 94%) as a white solid.

[0468] LC-MS: [M+H] + :251.1.

[0469] Synthesis of 2-cyclopropyl-6-isopropoxy-7-nitro-3,4-dihydroisoquinolin-1(2H)-one:

[0470] To a solution of 6-isopropoxy-7-nitro-3,4-dihydroisoquinolin-1(2H)-one (100 mg, 0.40 mmol) in 1,4-dioxane (10 mL) were added cyclopropylboronic acid (68.7 mg, 0.80 mmol), copper acetate (72.6 mg, 0.40 mmol), 4-dimethylaminopyridine (195.3 mg, 1.6 mmol), and pyridine (79.1 mg, 1.00 mmol), followed by reaction at 100°C under an oxygen atmosphere for 16 hours. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with aqueous sodium chloride solution (10 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether = 1 / 1) to afford product 38-3 (70 mg, 57% yield) as a white solid.

[0471] LC-MS: [M+H] + :291.1.

[0472] Synthesis of 7-amino-2-cyclopropyl-6-isopropoxy-3,4-dihydroisoquinolin-1(2H)-one:

[0473] To a solution of 2-cyclopropyl-6-isopropoxy-7-nitro-3,4-dihydroisoquinolin-1(2H)-one (30 mg, 0.10 mmol) in methanol (20 mL) was added palladium hydroxide on carbon (60.4 mg, 25% water content). The mixture was hydrogenated using a hydrogen balloon for 3 hours. The filtrate was then collected by filtration and concentrated under reduced pressure to afford the product 38-4 (25 mg, 96% yield) as a white solid.

[0474] LC-MS: [M+H] + :261.2.

[0475] Synthesis of 2-(chloromethyl)-3-(2-cyclopropyl-6-isopropoxy-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)quinazolin-4(3H)-one:

[0476] To a solution of 2-(2-chloroacetylamino)benzoic acid (40.0 mg, 0.19 mmol) in acetonitrile (20 mL) was added N,N-diisopropylethylamine (29.1 mg, 0.22 mmol). After stirring for 2 minutes, phosphorus trichloride (64.3 mg, 0.47 mmol) was added. After stirring for 10 minutes, a solution of 7-amino-2-cyclopropyl-6-isopropoxy-3,4-dihydroisoquinolin-1(2H)-one (48.8 mg, 0.19 mmol) in acetonitrile (2 mL) was added. The mixture was heated at reflux for 6 hours. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with aqueous sodium bicarbonate (20 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel chromatography (dichloromethane / methanol = 15 / 1) to give the product 38-5 (40 mg, yield: 50%) as a white solid.

[0477] LC-MS: [M+H] + :438.1.

[0478] Synthesis of 3-(2-cyclopropyl-6-isopropoxy-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-(piperazin-1-ylmethyl)quinazolin-4(3H)-one:

[0479] 2-(Chloromethyl)-3-(2-cyclopropyl-6-isopropoxy-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)quinazolin-4(3H)-one (40 mg, 0.09 mmol) and piperazine (23.6 mg, 0.27 mmol) were dissolved in tetrahydrofuran (20 mL), and the mixture was heated at reflux for 16 hours. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with aqueous sodium bicarbonate (10 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to afford 38-6 (35 mg crude product) as a white solid, which was used directly in the next reaction.

[0480] LC-MS: [M+H] + :488.3.

[0481] Synthesis of 2-((4-(2-(4-chlorophenoxy)acetyl)piperazin-1-yl)methyl)-3-(2-cyclopropyl-6-isopropoxy-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)quinazolin-4(3H)-one:

[0482] To a solution of 3-(2-cyclopropyl-6-isopropoxy-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-(piperazin-1-ylmethyl)quinazolin-4(3H)-one (35 mg, 0.07 mmol) in dichloromethane (20 mL) were added N,N-diisopropylethylamine (13.9 mg, 0.10 mmol) and 2-(4-chlorophenoxy)acetyl chloride (14.7 mg, 0.17 mmol). The mixture was stirred at room temperature for 6 hours. Aqueous sodium bicarbonate (20 mL) was added, followed by extraction with dichloromethane (30 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The crude product was purified by silica gel chromatography (petroleum ether / ethyl acetate = 1 / 1) to give a crude product, and then purified by preparative plate (petroleum ether / ethyl acetate = 1 / 1) to give compound 198 (19.6 mg, yield: 41%) as a white solid.

[0483] LC-MS: [M+H] + :656.2.

[0484] HPLC:98.08%.

[0485] 1H NMR: (400MHz, CD3OD) δ8.25(dd,J=8.0,1.2Hz,1H),8.02(s,1H),7.94-7.89(m,1H),7.80(d,J=8.0Hz,1H),7.64-7.59(m,1H),7.29-7.25(m,2H), 7.13(s,1H),6.96-6.93(m,2H),4.80-4.75(m,3H),3.72-3.64(m,2H),3.60-3.35(m,6H),3.10-3.08(m,2H),2.88-2.83(m,1 H),2.64-2.40(m,2H),2.34-2.07(m,2H),1.25(d,J=6.0Hz,3H),1.19(d,J=6.0Hz,3H),0.95-0.90(m,2H),0.81-0.77(m,2H).

[0486] Example 39 / 40

[0487] Compound 198 (17.6 mg) was chiral separated by SFC (chromatographic column: OJ-H 250 mm × 20 mm, 5 μm, mobile phase: carbon dioxide-methanol (NH4OH 0.2%) gradient: 0-40%) to obtain the title compound (5.2 mg, 5.5 mg)

[0488] Single configuration compound (shorter retention time)

[0489] It was analyzed by chiral analysis chromatography conditions, RT = 2.11min

[0490] LC-MS: [M+H] + :656.2.

[0491] HPLC: 100%.

[0492] 1H NMR: (400MHz, CD3OD) δ8.25 (dd, J=8.0, 1.2Hz, 1H), 8.02 (s, 1H), 7.94-7.89 (m, 1H), 7.80 (d, J=8.0 Hz,1H),7.64-7.59(m,1H),7.29-7.25(m,2H),7.13(s,1H),6.96-6.93(m,2H),4.80-4.75(m,3H),3 .72-3.64(m,2H),3.60-3.35(m,6H),3.10-3.08(m,2H),2.88-2.83(m,1H),2.64-2.40(m,2H),2.3 4-2.07(m,2H),1.25(d,J=6.0Hz,3H),1.19(d,J=6.0Hz,3H),0.95-0.90(m,2H),0.81-0.77(m,2H).

[0493] Single configuration compound (longer retention time)

[0494] It was analyzed by chiral analysis chromatography conditions, RT = 4.27min

[0495] LC-MS: [M+H] + :656.2.

[0496] HPLC: 97.07%.

[0497] 1 H NMR: (400MHz, CD3OD) δ8.25(dd,J=8.0,1.2Hz,1H),8.02(s,1H),7.94-7.89(m,1H),7.80(d,J=8.0Hz,1H),7.64-7.59(m,1H),7.29-7.25(m,2H), 7.13(s,1H),6.96-6.93(m,2H),4.80-4.75(m,3H),3.72-3.64(m,2H),3.60-3.35(m,6H),3.10-3.08(m,2H),2.88-2.83(m,1 H),2.64-2.40(m,2H),2.34-2.07(m,2H),1.25(d,J=6.0Hz,3H),1.19(d,J=6.0Hz,3H),0.95-0.90(m,2H),0.81-0.77(m,2H).

[0498] Example 41

[0499] Referring to Example 20, 41-1 was substituted for 1-4 to obtain the target compound 204 (7 mg)

[0500] LC-MS: [M+H]+: 669.0.

[0501] HPLC:99.79%.

[0502] 1H NMR: (400MHz, CD3OD) δ8.30(d,J=8.2Hz,1H),7.94-7.89(m,1H),7.80-7.76(m,1H),7.70(d,J=8.0Hz, 1H),7.58-7.54(m,1H),7.28-7.26(m,2H),7.09(s,1H),6.97-6.91(m,2H),6.84-6.83(m,1H),4.81-4 .71(m,3H),3.75-3.62(m,3H),3.60-3.41(m,4H),3.11-3.02(m,3H),2.96-2.81(m,3H),2.78-2.70(m ,1H),1.27-1.16(m,6H),0.94-0.90(m,2H),0.79-0.77(m,3H),0.63-0.62(m,1H),0.56-0.55(m,1H).

[0503] Example 42

[0504] Referring to Example 1, cyclopropanol was substituted for isopropanol 1-2 to obtain the target compound 6 (30 mg)

[0505] LC-MS: [M+H] + :645.0.

[0506] HPLC: 98.9%.

[0507] 1 H NMR(400MHz,MeOD)δ8.23(d,J=7.0Hz,1H),7.90-7.88(m,2H),7.76(d,J=8.0H z,1H),7.63-7.56(m,2H),7.28-7.26(m,2H),6.96-6.94(m,2H),4.81-4.71(m, 2H),4.05-4.03(m,1H),3.82-3.70(m,1H),3.64-3.62(m,2H),3.30-3.04(m,2H ),2.79-2.74(m,2H),2.33-2.28(m,2H),0.86-0.84(m,2H),0.77-0.60(m,5H).

[0508] Example 43 / 44

[0509] Compound 6 (30 mg) was chiral separated by SFC (chromatographic column: OJ-H 250 mm × 20 mm, 5 μm, mobile phase: carbon dioxide-methanol (NH4OH 0.2%) gradient: 0-40%) to obtain the title compound (13.1 mg, 12.9 mg)

[0510] Single configuration compound (shorter retention time)

[0511] It was analyzed by chiral analysis chromatography conditions, RT = 2.05min

[0512] LC-MS: [M+H] + :645.0.

[0513] HPLC: 98.9%.

[0514] 1 H NMR(400MHz,MeOD)δ8.23(d,J=7.0Hz,1H),7.90-7.88(m,2H),7.76(d,J=8.0H z,1H),7.63-7.56(m,2H),7.28-7.26(m,2H),6.96-6.94(m,2H),4.81-4.71(m, 2H),4.05-4.03(m,1H),3.82-3.70(m,1H),3.64-3.62(m,2H),3.30-3.04(m,2H ),2.79-2.74(m,2H),2.33-2.28(m,2H),0.86-0.84(m,2H),0.77-0.60(m,5H).

[0515] Single configuration compound (longer retention time)

[0516] It was analyzed by chiral analysis chromatography conditions, RT = 2.81min

[0517] LC-MS: [M+H] + :645.0.

[0518] HPLC: 95.9%.

[0519] 1H NMR(400MHz,MeOD)δ8.23(d,J=7.0Hz,1H),7.90-7.88(m,2H),7.76(d,J=8.0H z,1H),7.63-7.56(m,2H),7.28-7.26(m,2H),6.96-6.94(m,2H),4.81-4.71(m, 2H),4.05-4.03(m,1H),3.82-3.70(m,1H),3.64-3.62(m,2H),3.30-3.04(m,2H ),2.79-2.74(m,2H),2.33-2.28(m,2H),0.86-0.84(m,2H),0.77-0.60(m,5H).

[0520] Example 45 / 46

[0521] Compound 1 (40 mg) was chiral separated by SFC (chromatographic column: WHELK-O1 250 mm × 21.1 mm, 5 μm, mobile phase: carbon dioxide-methanol (NH4OH 0.2%) gradient: 0-40%) to obtain the title compound (6.9 mg, 8.1 mg)

[0522] Single configuration compound (shorter retention time)

[0523] It was analyzed by chiral analysis chromatography conditions, RT = 8.7min

[0524] LC-MS: [M+H] + :615.2.

[0525] HPLC: 99.74%

[0526] 1 HNMR (400MHz, CD3OD) δ8.25(d,J=7.9Hz,1H),7.96-7.85(m,3H),7.81(s,1H),7.63(t,J=7.5Hz,1H),7.46(d,J=8.9Hz,1H),7.27-7.22(m,2H) ,6.97-6.91(m,2H),4.86-4.79(m,4H),4.32-4.27(m,1H),3.95-3.91( m,4H),3.56-3.34(m,4H),1.26(d,J=6.0Hz,3H),1.16(d,J=6.0Hz,3H).

[0527] Single configuration compound (longer retention time)

[0528] It was analyzed by chiral analysis chromatography conditions, RT = 11.45min

[0529] LC-MS: [M+H] + :615.2.

[0530] HPLC: 99.66%

[0531] 1 HNMR (400MHz, CD3OD) δ8.25(d,J=7.9Hz,1H),7.96-7.85(m,3H),7.81(s,1H),7.63(t,J=7.5Hz,1H),7.46(d,J=8.9Hz,1H),7.27-7.22(m,2H) ,6.97-6.91(m,2H),4.86-4.79(m,4H),4.32-4.27(m,1H),3.95-3.91( m,4H),3.56-3.34(m,4H),1.26(d,J=6.0Hz,3H),1.16(d,J=6.0Hz,3H).

[0532] Example 47

[0533] Synthesis of 3-isopropoxy-6-(trifluoromethyl)pyridin-2-amine:

[0534] To a solution of 2-bromo-3-isopropoxy-6-(trifluoromethyl)pyridine (500 mg, 1.75 mmol) in ethylene glycol (5 mL) and aqueous ammonia (5 mL) were added cuprous oxide (25 mg, 0.175 mmol), DMEDA (15.46 mg, 0.175 mmol), and potassium carbonate (242.4 mg, 1.75 mmol). The mixture was stirred in a sealed container at 80°C for 15 hours. The reaction was quenched by the addition of water (20 mL). Extraction was performed with ethyl acetate (20 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate and concentrated in vacuo to yield the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 2:1) to afford propoxy-6-(trifluoromethyl)pyridin-2-amine (120 mg, yield: 27.8%) as a colorless liquid.

[0535] LC-MS: [M+H] + :221.0.

[0536] Synthesis of 2-(chloromethyl)-3-(3-isopropoxy-6-(trifluoromethyl)pyridin-2-yl)quinazolin-4(3H)-one:

[0537] To a solution of 2-(2-chloroacetamido)benzoic acid (100 mg, 0.468 mmol) in acetonitrile (10 mL) was added DIEA (72.6 mg, 0.561 mmol). The mixture was stirred at 25°C for 5 minutes, followed by the addition of phosphorus trichloride (160.7 mg, 1.17 mmol). The mixture was stirred at 25°C for 10 minutes. 3-Isopropoxy-6-(trifluoromethyl)pyridin-2-amine (103.5 mg, 0.468 mmol) was then added, and the mixture was stirred at 80°C for 15 hours. The reaction was quenched by the addition of water (20 mL). The mixture was extracted with ethyl acetate (20 mL x 2). The combined organic layers were dried over anhydrous sodium sulfate and concentrated in vacuo to yield the crude product. The crude product was purified by thin-layer chromatography (petroleum ether:ethyl acetate = 3:1) to afford 47-3 (90 mg, 43.9% yield) as a white solid.

[0538] LC-MS: [M+H] + :398.0.

[0539] Synthesis of (S)-tert-butyl 4-((3-(3-isopropoxy-6-(trifluoromethyl)pyridin-2-yl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-3-methylpiperazine-1-carboxylate:

[0540] To a solution of 2-(chloromethyl)-3-(3-isopropoxy-6-(trifluoromethyl)pyridin-2-yl)quinazolin-4(3H)-one (100 mg, 0.25 mmol) in acetonitrile (20 mL) was added tert-butyl-(S)-3-methylpiperazine-1-carboxylate (50.4 mg, 0.25 mmol), potassium carbonate (51.9 mg, 0.376 mmol), and potassium iodide (41.6 mg, 0.25 mmol). The mixture was stirred at 80°C for 5 hours. Water (20 mL) was added for dilution. The mixture was extracted with ethyl acetate (20 mL x 2). The combined organic layers were dried over anhydrous sodium sulfate and concentrated in vacuo to yield the crude product. The crude product was purified by thin-layer chromatography (petroleum ether:ethyl acetate = 3:1) to afford 47-4 (110 mg, 62.2% yield) as a white solid.

[0541] LC-MS: [M+H] + :562.0.

[0542] Synthesis of (S)-3-(3-isopropoxy-6-(trifluoromethyl)pyridin-2-yl)-2-((2-methylpiperazin-1-yl)methyl)quinazolin-4(3H)-one:

[0543] To a solution of 47-4 (110 mg, 0.195 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (1 mL), and the mixture was stirred at 25° C. for 15 hours. The mixture was concentrated in vacuo to give the crude product 47-5 (90 mg, yield: 69.7%), which was used directly in the next reaction.

[0544] LC-MS: [M+H] + :462.1.

[0545] Synthesis of (S)-2-((4-(2-(4-chlorophenoxy)acetyl)-2-methylpiperazin-1-yl)methyl)-3-(3-isopropoxy-6-(trifluoromethyl)pyridin-2-yl)quinazolin-4(3H)-one:

[0546] To a dichloromethane solution (5 mL) of 47-5 (80 mg, 0.173 mmol) were added 2-(4-chlorophenoxy)acetyl chloride (35.4 mg, 0.173 mmol) and DIEA (44.7 mg, 0.346 mmol), and the mixture was stirred at 25°C for 15 hours. The organic phase was concentrated in vacuo to obtain a crude product. The crude product was purified by HPLC (ACN-H2O-(0.1% TFA), 20-30%) to give compound 207 (31.8 mg, yield: 28.2%).

[0547] LC-MS: [M+H] + :630.0.

[0548] HPLC: 96.7%.

[0549] 1 H NMR (400MHz, CD3OD) δ8.24(d,J=8.0Hz,1H),8.04-7.95(m,1H),7.89-7.78(m,3H),7.62(t,J=7.6Hz,1H),7.28-7.16(m,2H),6.90-6.85(m, 2H),4.87-4.84(m,1H),4.76-4.74(m,2H),4.25-3.80(m,2H),3.73-3 .30(m,3H),3.22-2.36(m,4H),1.32-1.22(m,6H),1.17-0.82(m,3H).

[0550] Example 48

[0551] Synthesis of tert-butyl 3-formyl-5,6-dihydroimidazo[1,5-a]pyrazine-7(8H)-carboxylate:

[0552] To a solution of tert-butyl 5,6-dihydroimidazo[1,5-a]pyrazine-7(8H)-carboxylate (200.0 mg, 0.89 mmol) in tetrahydrofuran (10 mL) at -78°C was slowly added n-butyl lithium (0.54 mL, 1.33 mmol, 2.5 Min Hexane). After stirring at -78°C for 1 hour, N,N-dimethylformamide (130.4 mg, 1.78 mmol) was added, and the mixture was warmed to room temperature and stirred for 16 hours. The mixture was quenched with saturated aqueous ammonium chloride (5 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with saturated sodium chloride solution (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate / petroleum ether = 1 / 1) to afford product 48-2 (170 mg, yield: 65%) as a white solid.

[0553] LC-MS: [M+H] + :252.1.

[0554] Synthesis of tert-butyl 3-(hydroxymethyl)-5,6-dihydroimidazo[1,5-a]pyrazine-7(8H)-carboxylate:

[0555] To a solution of 48-2 (170 mg, 0.67 mmol) in methanol (20 mL) was added sodium borohydride (51.0 mg, 1.35 mmol) at 0°C, and then the mixture was allowed to react at room temperature for 16 hours. The mixture was concentrated under reduced pressure, and the residue was added with saturated aqueous sodium chloride solution (20 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel chromatography (dichloromethane / methanol = 15 / 1) to give the product 48-3 (70 mg, yield: 39%) as a white solid.

[0556] LC-MS: [M+H] + :254.1.

[0557] Synthesis of tert-butyl 3-((4-chlorophenoxy)methyl)-5,6-dihydroimidazo[1,5-a]pyrazine-7(8H)-carboxylate:

[0558] To a solution of 48-3 (60.0 mg, 0.24 mmol) in tetrahydrofuran (20 mL) at 0°C under nitrogen was added p-chlorophenol (30.4 mg, 0.24 mmol), triphenylphosphine (92.9 mg, 0.35 mmol), and diisopropyl azodicarboxylate (71.6 mg, 0.35 mmol), followed by warming to room temperature for 16 hours. The mixture was quenched with saturated aqueous sodium chloride (20 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The crude product was purified by silica gel chromatography (dichloromethane / methanol = 15 / 1) to afford 48-4 (50 mg crude) as a white solid, which was used directly in the next reaction.

[0559] LC-MS: [M+H] + :364.0.

[0560] Synthesis of 3-((4-chlorophenoxy)methyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine:

[0561] To a solution of 48-4 (50.0 mg, 0.12 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (2 mL), stirred at room temperature for 3 hours, and concentrated under reduced pressure to give a white solid 48-5 (60 mg crude) which was used directly in the next reaction.

[0562] LC-MS: [M+H] + :264.1.

[0563] Synthesis of 2-((3-((4-chlorophenoxy)methyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)methyl]-3-(2-isopropoxy-5-(trifluoromethyl)phenyl)quinazolin-4(3H)-one:

[0564] To a solution of 48-5 (30 mg, 0.11 mmol) in acetonitrile (20 mL) were added 2-(chloromethyl)-3-(2-isopropoxy-5-(trifluoromethyl)phenyl)quinazolin-4(3H)-one 18-1 (45.3 mg, 0.11 mmol), potassium carbonate (62.9 mg, 0.45 mmol) and potassium iodide (18.9 mg, 0.11 mmol), and the mixture was heated at reflux for 16 hours. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was diluted with aqueous sodium bicarbonate (10 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel chromatography (petroleum ether / ethyl acetate = 1 / 1) to give the crude product, which was then purified by high performance liquid chromatography to give compound 208 (6.1 mg, 8%) as a white solid.

[0565] LC-MS: [M+H] + :623.9.

[0566] HPLC:97.96%.

[0567] 1 H NMR: (400MHz, CD3OD) δ8.25(d,J=8.0Hz,1H),7.94-7.90(m,1H),7.83-7.77(m,3H),7.64-7.60(m,1H),7.33-7.29(m,2H),7.16(d,J=9.4Hz, 1H),7.07-7.03(m,2H),6.68(s,1H),5.12(s,2H),4.47-4.43(m,1H),4.06-4.00(m,1H),3.81-3.79(m,1H ),3.70-3.60(m,2H),3.57-3.50(m,2H),2.75-2.71(m,2H),1.13(d,J=6.0Hz,3H),1.03(d,J=6.0Hz,3H).

[0568] Example 49 / 50

[0569] Compound 208 (4.1 mg) was chiral separated by SFC (chromatographic column: IC 250 mm × 20 mm, 5 μm, mobile phase: carbon dioxide-methanol (NH4OH 0.2%) gradient: 0-40%) to obtain the title compound (1.2 mg, 0.9 mg)

[0570] Single configuration compound (shorter retention time)

[0571] It was analyzed by chiral analysis chromatography conditions, RT = 4.03min

[0572] LC-MS: [M+H] + :623.9.

[0573] HPLC:88.23%.

[0574] 1 H NMR: (400MHz, CD3OD) δ8.25 (d, J=8.0Hz, 1H), 7.94-7.90 (m, 1H), 7.83-7.77 (m, 3H), 7.6 4-7.60(m,1H),7.33-7.29(m,2H),7.16(d,J=9.4Hz,1H),7.07-7.03(m,2H),6.68(s,1H ),5.12(s,2H),4.47-4.43(m,1H),4.06-4.00(m,1H),3.81-3.79(m,1H),3.70-3.60(m, 2H),3.57-3.50(m,2H),2.75-2.71(m,2H),1.13(d,J=6.0Hz,3H),1.03(d,J=6.0Hz,3H).

[0575] Single configuration compound (longer retention time)

[0576] It was analyzed by chiral analysis chromatography conditions, RT = 4.44min

[0577] LC-MS: [M+H] + :623.9.

[0578] HPLC: 95.26%.

[0579] 1 H NMR: (400MHz, CD3OD) δ8.25 (d, J=8.0Hz, 1H), 7.94-7.90 (m, 1H), 7.83-7.7 7(m,3H),7.64-7.60(m,1H),7.33-7.29(m,2H),7.16(d,J=9.4Hz,1H),7.07 -7.03(m,2H),6.68(s,1H),5.12(s,2H),4.47-4.43(m,1H),4.06-4.00(m,1 H),3.81-3.79(m,1H),3.70-3.60(m,2H),3.57-3.50(m,2H),2.75-2.71(m, 2H), 1.13 (d, J=6.0Hz, 3H), 1.03 (d, J=6.0Hz, 3H).

[0580] Example 51

[0581] Synthesis of tert-butyl 3-hydroxymethyl-1-methyl-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxylate:

[0582] To a solution of 6-tert-butyl 3-ethyl 2-methyl-1,4,5,7-tetrahydropyrazolo[3,4-c]pyridine-3,6-dicarboxylate 51-1 (65 mg, 0.22 mmol) in tetrahydrofuran (5 mL) was added lithium aluminum hydride (17.5 mg, 0.44 mmol) at 0°C. The mixture was stirred at room temperature for 4 hours. The reaction solution was quenched with saturated ammonium chloride solution. The reaction solution was diluted with ethyl acetate (50 mL) and washed with water (100 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude product 51-2 (65 mg) as a clear oil, which was used directly in the next reaction.

[0583] LC-MS: [M+H] + :268.1.

[0584] Synthesis of tert-butyl 3-chloromethyl-1-methyl-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxylate:

[0585] To a solution of 51-2 (65 mg, 0.24 mmol) in dichloromethane (10 mL) was added thionyl chloride (43 mg, 0.36 mmol) at room temperature. The mixture was stirred at 40°C for 6 hours. The reaction solution was diluted with dichloromethane (50 mL) and washed with water (100 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude product 51-3 (51 mg, yield: 51.4%) as a yellow oil, which was used directly in the next reaction.

[0586] LC-MS: [M+H] + :286.2.

[0587] Synthesis of tert-butyl 3-((4-chlorophenoxy)methyl)-1-methyl-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxylate:

[0588] To a solution of 51-3 (50 mg, 0.17 mmol) in acetonitrile (5 mL) was added 4-chlorophenol (27 mg, 0.21 mmol) and potassium carbonate (36 mg, 0.26 mmol) at room temperature. The mixture was stirred at 80°C for 16 hours. The reaction solution was concentrated under reduced pressure and diluted with ethyl acetate (50 mL) and washed with water (100 mL) and saturated brine (100 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (dichloromethane:methanol = 10:1) to obtain product 51-4 (70 mg, yield: 84.7%).

[0589] LC-MS: [M+H] + :378.1.

[0590] Synthesis of 3-((4-chlorophenoxy)methyl)-1-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine:

[0591] To a solution of 51-4 (66 mg, 0.17 mmol) in dichloromethane (10 mL) was added trifluoroacetic acid (2 mL) at room temperature. The mixture was stirred at room temperature for 2 hours. The solution was concentrated under vacuum to afford the product 51-5 (70 mg crude product), which was used directly in the next reaction.

[0592] LC-MS: [M+H] + :278.2.

[0593] Synthesis of 2-((3-((4-chlorophenoxy)methyl)-1-methyl-1,4,5,7-tetrahydro-6H-pyrazolo[34-c]pyridin-6-yl)methyl)-3-(2-isopropoxy-5-(trifluoromethyl)phenyl)quinazolin-4(3H)-one:

[0594] To a solution of 3-((4-chlorophenoxy)methyl)-1-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine 51-5 (70 mg, 0.18 mmol) in acetonitrile (10 mL) was added 18-1 (49 mg, 0.18 mmol), potassium iodide (38 mg, 0.23 mmol), and potassium carbonate (49 mg, 0.35 mmol) at room temperature. The mixture was stirred at 80°C for 16 hours. The reaction solution was concentrated under reduced pressure and diluted with ethyl acetate (50 mL). The organic phase was washed with water (300 mL) and saturated brine (100 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product. The crude product was purified by high-performance liquid chromatography (acetonitrile-water (0.1% formic acid) gradient: 50-70) to obtain compound 214 (19.57 mg, yield: 17.6%).

[0595] LC-MS: [M+H] + :638.3.

[0596] HPLC: 99.8%

[0597] 1 HNMR(400MHz,CD3OD)δ8.26-8.24(m,1H),7.91-7.84(m,1H),7.79-7.68(m,3H),7 .57(t,J=7.6Hz,1H),7.26-7.19(m,2H),7.12(d,J=8.8Hz,1H),6.96-6.87(m,2H) ,5.00(s,2H),4.45-4.41(m,1H),3.64-3.59(m,1H),3.59(s,3H),3.44-3.40(m,1 H),3.34(s,2H),2.53-2.19(m,4H),1.31(d,J=6.0Hz,3H),1.13(d,J=6.0Hz,3H).

[0598] Example 52 / 53

[0599] Compound 214 (17 mg) was chiral separated by SFC (chromatographic column: AD-H 250 mm × 20 mm, 5 μm, mobile phase: carbon dioxide-ethanol (NH4OH 0.2%) gradient: 0-40%) to obtain the title compound (4.6 mg, 4.8 mg)

[0600] Single configuration compound (shorter retention time)

[0601] It was analyzed by chiral analysis chromatography conditions, RT = 2.43min

[0602] LC-MS: [M+H] + :638.3.

[0603] HPLC: 99.8%

[0604] 1HNMR(400MHz,CD3OD)δ8.26-8.24(m,1H),7.91-7.84(m,1H),7.79-7.68(m,3H),7 .57(t,J=7.6Hz,1H),7.26-7.19(m,2H),7.12(d,J=8.8Hz,1H),6.96-6.87(m,2H) ,5.00(s,2H),4.45-4.41(m,1H),3.64-3.59(m,1H),3.59(s,3H),3.44-3.40(m,1 H),3.34(s,2H),2.53-2.19(m,4H),1.31(d,J=6.0Hz,3H),1.13(d,J=6.0Hz,3H).

[0605] Single configuration compound (longer retention time)

[0606] It was analyzed by chiral analysis chromatography conditions, RT = 3.15min

[0607] LC-MS: [M+H] + :638.3.

[0608] HPLC: 99.8%

[0609] 1 HNMR(400MHz,CD3OD)δ8.26-8.24(m,1H),7.91-7.84(m,1H),7.79-7.68(m,3H),7 .57(t,J=7.6Hz,1H),7.26-7.19(m,2H),7.12(d,J=8.8Hz,1H),6.96-6.87(m,2H) ,5.00(s,2H),4.45-4.41(m,1H),3.64-3.59(m,1H),3.59(s,3H),3.44-3.40(m,1 H),3.34(s,2H),2.53-2.19(m,4H),1.31(d,J=6.0Hz,3H),1.13(d,J=6.0Hz,3H).

[0610] Example 54 / 55

[0611] Compound 17 (14 mg) was chiral separated by SFC (chromatographic column: WHELK-O1 250 mm × 21.1 mm, 5 μm, mobile phase: carbon dioxide-methanol (NH4OH 0.2%) gradient: 0-40%) to obtain the title compound (2.9 mg, 3.5 mg)

[0612] Single configuration compound (shorter retention time)

[0613] It was analyzed by chiral analysis chromatography conditions, RT = 8.84min

[0614] LC-MS: [M+H] + :628.9.

[0615] HPLC: 99.7%

[0616] 1 H NMR (400MHz, CD3OD) δ8.24(d,J=8.0Hz,1H),7.84-7.82(m,2H),7.72(s,1H),7.62(t,J=7.2Hz,1H),7.28-7.22(m,3H),6.93-6.90(m,2H ),4.82-4.75(m,3H),3.98-3.94(m,1H)3.86-3.44(m,5H),3.25-2.88(m,4H),2.57(s,3H),1.25(d,J=6.0Hz,3H),1.15(d,J=6.0Hz,3H).

[0617] Single configuration compound (longer retention time)

[0618] It was analyzed by chiral analysis chromatography conditions, RT = 11.45min

[0619] LC-MS: [M+H] + :628.9.

[0620] HPLC: 99.1%

[0621] 1 H NMR (400MHz, CD3OD) δ8.24(d,J=8.0Hz,1H),7.84-7.82(m,2H),7.72(s,1H),7.62(t,J=7.2Hz,1H),7.28-7.22(m,3H),6.93-6.90(m,2H ),4.82-4.75(m,3H),3.98-3.94(m,1H)3.86-3.44(m,5H),3.25-2.88(m,4H),2.57(s,3H),1.25(d,J=6.0Hz,3H),1.15(d,J=6.0Hz,3H).

[0622] In the following embodiments, the IKE structure is as follows:

[0623] Example 56 Activity Test of Compounds for Inhibiting Glutamate Release

[0624] The following is an in vitro assay for the inhibition of glutamate release by a compound. This assay can be used to evaluate or characterize the inhibitory activity of the compounds of the present invention as SLC7A11 inhibitors.

[0625] 1. Cell plating:

[0626] 1) Discard the culture medium from the T75 flask, add 1 ml of PBS and wash once, then aspirate and discard the PBS.

[0627] 2) Add 1 ml of 0.25% trypsin and incubate at 37°C for 1 min. Observe the cells under a microscope to see if they are rounding or falling off.

[0628] 3) Add 1 ml of complete culture medium to terminate digestion, add 2 ml of culture medium to a 15 ml centrifuge tube and centrifuge (1000 rpm, 3 min);

[0629] 4) After aspirating the supernatant, add 1 ml of complete medium and gently pipette to mix. Take 10 μl and add it to 190 μl of complete medium (diluted 20-fold), gently pipette to mix and count.

[0630] 5) According to the counting results, dilute the cells to 6x10 5 / ml, vortex mix each tube before plating, then add to the separation tank, spread the plate with a dispenser, 100ul / well;

[0631] 2. Compound Treatment and Fluorescence Detection

[0632] 1) 60,000 cells / well of a 96-well plate with a cell density of >95% confluency. 24 hours after plating the cells, discard the culture medium and wash the cells three times with EBSS (Earle's Balanced Salt Solution) to remove the glutamine-containing culture medium.

[0633] 2) Compound preparation: Add 10 ml of EBSS to a 15 ml centrifuge tube, add 100 μl of 8 mM cystein (100x), vortex to mix, remove 1 ml as the total group, use this cystein-containing EBSS to prepare 500 μl of 10 μM compound (vortex to mix after preparation), add 0.5‰ of DMSO to the remaining cystein-containing EBSS, and then dilute it 3-fold to 8 concentration gradients (i.e., 10.000, 3.333, 1.111, 0.370, 0.123, 0.041, 0.014, 0.005, DMSO), add the drug, 100 μl / well, and incubate at 37°C for 4 h.

[0634] 3) Remove 90 μl of culture medium from each well using a pipette and add 45 μl of Tris buffer (100 mM, pH 7.4) containing glutamate oxidase (0.02 U / mL), horseradish peroxidase (0.125 U / mL), and Amplex UltraRed (10 μM). Measure fluorescence after 15 minutes (Ex 530, Em 590, with shaking).

[0635] 4) Process the data according to the following formula:

[0636] Inhibition Rate (%)=1-(Unknown-Blank)) / (Totals-Blanks))*100.

[0637] IC of compounds inhibiting glutamate release 50 The (nM) values ​​are shown in Table 1.

[0638] Table 1

[0639] Example 57 Compound Inhibits Viability of HT1080 Cells

[0640] 1. Compound dissolution: Dissolve in DMSO to a 20 mM stock solution

[0641] 2. Cell plating:

[0642] 1) Discard the culture medium from the T75 flask, add 2 ml of PBS and wash once, then aspirate and discard the PBS.

[0643] 2) Add 1 ml of 0.25% trypsin and incubate at 37°C for 1 min. Observe the cells under a microscope to see if they are rounding or falling off.

[0644] 3) Add 1 ml of complete culture medium to terminate digestion, add 2 ml of culture medium to a 15 ml centrifuge tube and centrifuge (1000 rpm, 3 min);

[0645] 4) After aspirating the supernatant, add 1 ml of complete medium and gently pipette to mix. Take 10 μl and add it to 190 μl of complete medium (diluted 20-fold), gently pipette to mix and count.

[0646] 5) According to the counting results, dilute the cells to 1.5x10 5 (Take 5 plates of cells as an example: divide the required cells into 5 15ml centrifuge tubes, fill up to 10ml / tube with complete medium, vortex mix each tube before plating, then add to the dispensing tank, and plate with a dispenser at 100ul / well, ensuring that the time taken to plate each plate is consistent and does not exceed 30s).

[0647] 6) Label the 96-well plates in the order of plating: plate 1, plate 2, plate 3, plate 4, plate 5.

[0648] 3. Drug preparation:

[0649] 1) Taking a 96-well plate as an example, prepare two 15 ml centrifuge tubes. Tube 1 contains 6 ml complete medium + 0.1% DMSO, and tube 2 contains 6 ml complete medium + 0.1% Fer-1; tube 3 contains 1 ml complete medium with 0.6% DMSO.

[0650] 2) Prepare 600 μl of stock solution (100 μM) containing 0.5% compound;

[0651] 3) After the preparation is completed, add 0.5% DMSO to tubes 1 and 2 in step 1;

[0652] 4) Prepare 2 x 9 1.5ml tubes for each compound. Add 400ul of the medium from tube 1 to 8 tubes and 400ul of the medium from tube 2 to the other 8 tubes. Take 200ul of the compound stock solution and add it to each of the 9 dilution tubes in sequence [vortex each tube before adding to the next tube].

[0653] 5) Add the prepared compounds to the corresponding wells of the 96-well plate. (The PBS in the edge wells also needs to be quantified to 100ul)

[0654] 6) Mark the laying time on each board

[0655] CCK8 detection

[0656] 1. Add 10ul of CCK8 to each 96-well plate using a 10ul single-pipette syringe in the order marked on the 96-well plate (mark the time for adding CCK8 and keep it consistent on each plate). The time between plates also needs to be recorded and kept consistent.

[0657] 2. After 2 hours, take out the wells from the 96-well plate in order to test OD450 (before testing, observe whether there are bubbles. If there are, use a 1 ml syringe needle to puncture the bubbles).

[0658] In vitro determination of the IC of compounds inhibiting HT1080 cell activity 50 The (nM) values ​​are shown in Table 2.

[0659] Table 2:

[0660] Example 58 Compound Inhibits Viability of Pancreatic Cancer Su86.86 Cells

[0661] 1. Compound dissolution: Dissolve in DMSO to a 20 mM stock solution

[0662] 2. Cell plating:

[0663] 1) Discard the culture medium from the T75 flask, add 2 ml of PBS and wash once, then aspirate and discard the PBS.

[0664] 2) Add 1 ml of 0.25% trypsin and incubate at 37°C for 1 min. Observe the cells under a microscope to see if they are rounding or falling off.

[0665] 3) Add 1 ml of complete culture medium to terminate digestion, add 2 ml of culture medium to a 15 ml centrifuge tube and centrifuge (1000 rpm, 3 min);

[0666] 4) After aspirating the supernatant, add 1 ml of complete medium and gently pipette to mix. Take 10 μl and add it to 190 μl of complete medium (diluted 20-fold), gently pipette to mix and count.

[0667] 5) According to the counting results, dilute the cells to 1.5x10 5 (Take 5 plates of cells as an example: divide the required cells into 5 15ml centrifuge tubes, fill up to 10ml / tube with complete medium, vortex mix each tube before plating, then add to the dispensing tank, and plate with a dispenser at 100ul / well, ensuring that the time taken to plate each plate is consistent and does not exceed 30s).

[0668] 6) Label the 96-well plates in the order of plating: plate 1, plate 2, plate 3, plate 4, plate 5.

[0669] Drug preparation:

[0670] 7) Taking a 96-well plate as an example, prepare two 15 ml centrifuge tubes. Tube 1 contains 6 ml complete medium + 0.1% DMSO, and tube 2 contains 6 ml complete medium + 0.1% Fer-1; tube 3 contains 1 ml complete medium with 0.6% DMSO.

[0671] 8) Prepare 600 μl of stock solution (100 μM) containing 0.5% compound;

[0672] 9) After the preparation is complete, add 0.5% DMSO to tubes 1 and 2 in step 1;

[0673] 10) For each compound, prepare 2 x 9 1.5ml tubes. Add 400ul of the medium from tube 1 to 8 tubes, and 400ul of the medium from tube 2 to the other 8 tubes. Take 200ul of the compound stock solution and add it to each of the 9 dilution tubes in sequence [vortex each tube before adding to the next tube].

[0674] 11) Add the prepared compounds to the corresponding wells of a 96-well plate. (The PBS in the edge wells also needs to be quantified to 100ul)

[0675] 12) Mark the laying time on each board

[0676] CCK8 detection

[0677] 1. Add 10ul of CCK8 to each 96-well plate using a 10ul single-pipette syringe in the order marked on the 96-well plate (mark the time for adding CCK8 and keep it consistent on each plate). The time between plates also needs to be recorded and kept consistent.

[0678] 2. After 2 hours, take out the wells from the 96-well plate in order to test OD450 (before testing, observe whether there are bubbles. If there are, use a 1 ml syringe needle to puncture the bubbles).

[0679] In vitro determination of the IC50 activity of the compound in inhibiting pancreatic cancer Su86.86 cells 50 The (nM) values ​​are shown in Table 3.

[0680] Table 3

[0681] Example 59 Compounds Promote Bone Marrow-Derived Dendritic Cell (BMDC) Cell Evoked Cells

[0682] The experimental process is as follows:

[0683] 1) After euthanizing db / db mice, the tibia was removed to isolate bone marrow cells;

[0684] 2) Culture in medium containing 12.5 ng / mL GM-CSF and induce differentiation for 10 days;

[0685] 3) For differentiated BMDC cells, use CellTrace TM The kit was Far red labeled and set aside.

[0686] 4) Resuspend 10 million / mL Jurkat T cells in 1 mL 0.1% BSA solution and use CellTrace TM The kit was Violet labeled;

[0687] 5) The labeled Jurkat T cells were incubated in a medium containing 1 μM staurosporine for 3 hours to induce early cell apoptosis, and the apoptosis rate was measured before use;

[0688] 6) Far red-labeled BMDCs were mixed with apoptosis-induced Jurkat T cells at a ratio of 1:5, and treated with different concentrations of compounds (including compound 186, compounds 199 / 200, and compounds 184 / 185) and incubated at 37°C for 4 h;

[0689] 7) Transfer the cells to a V-bottom 96-well plate, wash the cells once with staining buffer, centrifuge at 350×g for 5 minutes, and stain with PI (1:100) for 15 minutes at room temperature.

[0690] 8) Wash twice with staining buffer, then resuspend the stained cells in 100 μL staining buffer and use the cells for FACS LSRFortessa analysis.

[0691] The results are shown in FIG1 . Compound 186, Compound 199 / 200, and Compound 184 / 185 have a significant effect of promoting BMDC efferocytosis and exhibit a good dose effect.

Claims

1. A compound represented by formula (I) or a deuterated substance thereof, or a pharmaceutically acceptable salt thereof: in, X is selected from: N, CR3; Z1 is independently selected from: C, N; Z2 is independently selected from: O, NR6, CR6; Dashed lines indicate no chemical bonds or single bonds; R1 is selected from: H, alkyl, alkenyl, alkynyl, cycloalkyl, alicyclic group optionally substituted by alkyl; R2 and R3 are each independently selected from: H, halogen, alkyl, cyano, haloalkyl, alkenyl, alkynyl, cycloalkyl, alicyclic optionally substituted by alkyl, -N(R6)R7, -OR8, -C(O)N(R9)R 10 、-P(O)(R9)R 10 , -SO2R'; R6 and R7 are each independently selected from: H, alkyl, haloalkyl, cycloalkyl, alicyclic optionally substituted by alkyl, alkanoyl, haloalkanoyl, cycloalkyl-aminoacyl, -SO2R', or, R6 and R7 are connected to each other and to the N to which they are connected to form alicyclic optionally substituted by alkyl; R8 is independently selected from the group consisting of: H, alkyl, haloalkyl, cycloalkyl, alicyclic optionally substituted by alkyl, alkanoyl, haloalkanoyl, heteroaryl; R9, R 10 Each is independently selected from: H, alkyl, haloalkyl, cycloalkyl, heterocyclic group optionally substituted by alkyl, or, when R9 and R 10 When connected to the same N or -P(O), R9 and R 10 It can also form an alicyclic group optionally substituted by an alkyl group together with the N or -P(O) to which it is commonly connected; Each R' is independently selected from: alkyl, haloalkyl, cycloalkyl, alicyclic, amino, alkylamino, cycloalkyl-amino; R4 is selected from: optionally substituted alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, alicyclic group, wherein the substituent in the optionally substituted alkyl is selected from: halogen, cyano, hydroxyl, alkoxy; R5 is selected from -CF3, acetyl; The number of substituents in the "alicyclic group optionally substituted by an alkyl group", "haloalkanoyl group", "haloalkyl group" and "optionally substituted alkyl group" is 1 or more, preferably 1, 2 or 3; Optionally, the compound represented by formula (I) is in the form of its tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture of these isomers.

2. The compound of formula (I) according to claim 1 or its deuterated form, or a pharmaceutically acceptable salt thereof: X is: CR3; Z1 is independently selected from: C, N; Z2 is independently selected from: O, NR6, CR6; Dashed lines indicate no chemical bonds or single bonds; R1 is selected from: H, alkyl, alkenyl, alkynyl, cycloalkyl, alicyclic group optionally substituted by alkyl; R2 and R3 are each independently selected from: H, halogen, alkyl, cyano, haloalkyl, alkenyl, alkynyl, cycloalkyl, alicyclic optionally substituted by alkyl, -N(R6)R7, -OR8, -C(O)N(R9)R 10 、-P(O)(R9)R 10 , -SO2R'; R6 and R7 are each independently selected from: H, alkyl, haloalkyl, cycloalkyl, alicyclic optionally substituted by alkyl, alkanoyl, haloalkanoyl, cycloalkyl-aminoacyl, -SO2R', or, R6 and R7 are connected to each other and to the N to which they are connected to form alicyclic optionally substituted by alkyl; R8 is independently selected from the group consisting of: H, alkyl, haloalkyl, cycloalkyl, alicyclic optionally substituted by alkyl, alkanoyl, haloalkanoyl, heteroaryl; R9, R 10 Each is independently selected from: H, alkyl, haloalkyl, cycloalkyl, heterocyclic group optionally substituted by alkyl, or, when R9 and R 10 When connected to the same N or -P(O), R9 and R 10 It can also form an alicyclic group optionally substituted by an alkyl group together with the N or -P(O) to which it is commonly connected; Each R' is independently selected from: alkyl, haloalkyl, cycloalkyl, alicyclic, amino, alkylamino, cycloalkyl-amino; R4 is selected from: optionally substituted alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, alicyclic, wherein, The substituent in the optionally substituted alkyl group is selected from the group consisting of: halogen, cyano, hydroxyl, alkoxy; R5 is selected from -CF3, acetyl; The number of substituents in the "alicyclic group optionally substituted by an alkyl group", "haloalkanoyl group", "haloalkyl group" and "optionally substituted alkyl group" is 1 or more, preferably 1, 2 or 3; Optionally, the compound represented by formula (I) is in the form of its tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture of these isomers.

3. The compound of formula (I) according to claim 1 or 2, or a deuterated substance thereof, or a pharmaceutically acceptable salt thereof, characterized in that: R5 is selected from Or R2 and R5 together with the benzene ring to which they are connected form the following structure: Each Ra is independently selected from alkyl, haloalkyl; Optionally, R5 is selected from 4. The compound of formula (I) according to any one of claims 1 to 3, or a deuterated substance thereof, or a pharmaceutically acceptable salt thereof, characterized in that: The halogen atoms in the "halogen", "haloalkyl" and "haloalkanoyl" are independently selected from F, Cl, Br and I; Optionally, the alkyl groups in the “alkyl”, “haloalkyl”, “alkoxy”, “alkanoyl”, “haloalkanoyl” and “aliphatic heterocyclic group optionally substituted by alkyl” are each independently C1-C 10 linear or branched alkyl; optionally, C1-C7 linear or branched alkyl; optionally, C1-C5 linear or branched alkyl; optionally, selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, isopentyl, 1-ethylpropyl, neopentyl, n-hexyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl ... hexyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 3-ethylpentyl or 2,2,3-trimethylbutyl; Optionally, the "alkenyl" is a C2-C 15 Straight or branched alkenyl, optional Ground, C2-C 10 Straight or branched alkenyl, optionally, C2-C8 straight or branched alkenyl, optionally, C2-C6 straight or branched alkenyl, optionally, C2-C4 straight or branched alkenyl, optionally, selected from ethenyl, propenyl, butenyl, pentenyl; Optionally, the "alkynyl" group contains one or more triple bonds. 15 A straight chain or branched alkynyl group, optionally, is C2-C 10 Straight chain or branched chain alkynyl, optionally, C2-C8 straight chain or branched chain alkynyl, optionally, C2-C6 straight chain or branched chain alkynyl, optionally, C2-C4 straight chain or branched chain alkynyl; optionally, selected from ethynyl, propynyl, hexynyl; Optionally, the cycloalkyl in the "cycloalkyl", "cycloalkyl-aminoacyl" and "cycloalkyl-amino" is C3-C 10 Monocyclic or bicyclic cycloalkyl, optionally, is C3-C7 monocyclic cycloalkyl, optionally, is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl; Optionally, the alicyclic group in the "alicyclic group", "alicyclic group optionally substituted by an alkyl group" and "alicyclic group-amino" is a 3-10-membered non-aromatic heterocycle containing 1, 2 or 3 heteroatoms selected from N, O and S on the ring; optionally, the alicyclic group is a 3-10-membered non-aromatic heterocycle containing 1 or 2 heteroatoms selected from N and O on the ring; optionally, the alicyclic group is a 3-6-membered non-aromatic heterocycle containing 1 or 2 heteroatoms selected from N and O on the ring; optionally, the alicyclic group is a 3-10-membered non-aromatic heterocycle containing 1 or 2 heteroatoms selected from N and S on the ring; optionally, the alicyclic group is a 3-10-membered non-aromatic heterocycle containing 1 or 2 heteroatoms selected from N and S on the ring; The heterocycle is a 3-6 membered non-aromatic heterocycle containing 1 or 2 heteroatoms selected from N and S on the ring; optionally, the heterocyclic group in the "heterocyclic group", "heterocyclic group optionally substituted by alkyl" and "heterocyclic group-amino" is selected from methyl-substituted azetidinyl, methyl-substituted azetidinyl, methyl-substituted azetidinyl, methyl-substituted azetidinyl, azetidinyl, azetidinyl, oxolanyl, oxolanyl, azetidinyl, piperidinyl, piperazinyl and alkenylpiperidine; optionally, the heterocyclic group in the "heterocyclic group", "heterocyclic group optionally substituted by alkyl" and "heterocyclic group-amino" is selected from Optionally, the heteroaromatic ring of the "heteroaryl" is a 5-10 membered heteroaromatic ring containing 1-2 heteroatoms selected from N and O on the ring; Optionally, the heteroaromatic group is selected from pyridyl, pyrrolyl, pyrazolyl, thiazolyl, oxazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, naphthyl, quinolyl, quinazolinyl; Optionally, The heteroaryl group is selected from pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, and pyrimidin-6-yl.

5. The compound of formula (I) according to any one of claims 1 to 4, or a deuterated substance thereof, or a pharmaceutically acceptable salt thereof, characterized in that: Selected from the following compounds:

6. The compound of formula (I) according to any one of claims 1 to 5, or a deuterated substance thereof, or a pharmaceutically acceptable salt thereof, characterized in that: The pharmaceutically acceptable salts include anionic salts and cationic salts of the compound represented by formula (I) or its deuterated product; Optionally, the pharmaceutically acceptable salt includes an alkali metal salt, an alkaline earth metal salt, or an ammonium salt of the compound represented by formula (I) or its deuterated product; optionally, the alkali metal includes sodium, potassium, lithium, and cesium, and the alkaline earth metal includes magnesium, calcium, and strontium; Optionally, the pharmaceutically acceptable salt includes a salt formed by the compound represented by formula (I) or its deuterated product and an organic base; Optionally, the organic base includes trialkylamine, pyridine, quinoline, piperidine, imidazole, picoline, dimethylaminopyridine, dimethylaniline, N-alkylmorpholine, 1,5-diazabicyclo[4.3.0]nonene-5, 1,8-diazabicyclo[5.4.0]undecene-7, 1,4-diazabicyclo[2.2.2]octane; Optionally, the trialkylamine includes trimethylamine, triethylamine, N-ethyldiisopropylamine; Optionally, the N-alkylmorpholine includes N-methylmorpholine; Optionally, the pharmaceutically acceptable salt includes a salt formed by the compound represented by formula (I) or its deuterated product and an acid; Optionally, the acid includes an inorganic acid and an organic acid; optionally, the inorganic acid includes hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, and carbonic acid; optionally, the organic acid includes formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, citric acid, citric acid, tartaric acid, carbonic acid, picric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, glutamic acid, and pamoic acid.

7. A pharmaceutical composition comprising the compound of formula (I) or a deuterated product thereof according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, and a pharmaceutical excipient.

8. Use of the compound of formula (I) or its deuterated substance according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 7 in the preparation of an SLC7A11 inhibitor, or in the preparation of a medicament for treating tumor diseases such as pancreatic cancer, liver cancer, osteosarcoma, glioma, or chronic diseases such as fatty liver, cirrhosis, liver fibrosis, pulmonary fibrosis, diabetic skin lesions, or neurodegenerative diseases such as amyotrophic lateral sclerosis, Parkinson's disease, and Alzheimer's disease.

9. A drug for inhibiting SLC7A11 or treating tumor diseases such as pancreatic cancer, liver cancer, osteosarcoma, glioma, or treating chronic diseases such as fatty liver, cirrhosis, liver fibrosis, pulmonary fibrosis, diabetic skin lesions, or neurodegenerative diseases such as amyotrophic lateral sclerosis, Parkinson's disease, Alzheimer's disease, etc., comprising a compound of formula (I) according to any one of claims 1 to 6 or a deuterated substance thereof, or a pharmaceutically acceptable salt thereof.

10. A compound of formula (I) or a deuterated substance thereof according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 7, for inhibiting SLC7A11 or treating tumor diseases such as pancreatic cancer, liver cancer, osteosarcoma, glioma, or treating chronic diseases such as fatty liver, cirrhosis, liver fibrosis, pulmonary fibrosis, diabetic skin lesions, or neurodegenerative diseases such as amyotrophic lateral sclerosis, Parkinson's disease, and Alzheimer's disease.

11. A method for inhibiting SLC7A11 or treating tumor diseases such as pancreatic cancer, liver cancer, osteosarcoma, glioma, or treating chronic diseases such as fatty liver, cirrhosis, liver fibrosis, pulmonary fibrosis, diabetic skin lesions, or neurodegenerative diseases such as amyotrophic lateral sclerosis, Parkinson's disease, and Alzheimer's disease, comprising administering to a subject in need thereof an effective amount of a compound of formula (I) according to any one of claims 1 to 6 or a deuterated substance thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 7.

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