WEE1 inhibitors and their preparation and use

Abstract

The present invention relates to a compound represented by formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and use thereof in preparing a medicament for treating diseases associated with WEE1 activity.

Description

Technical Field

[0001] The present application relates to compounds that inhibit the activity of WEE1 kinase, in particular to compounds represented by formula (I), or stereoisomers thereof, or pharmaceutically acceptable salts thereof, as well as methods for preparing these compounds, pharmaceutical compositions thereof, and uses thereof in preparing drugs for treating diseases associated with WEE1 activity. Background Art

[0002] WEE1 tyrosine kinase is a checkpoint in the G2 phase of the cell cycle. The cell cycle is tightly regulated. When the cell's DNA is not damaged, the G1 phase, S phase, and G2 phase checkpoints promote the cell to enter the division phase and ensure the smooth completion of the cell cycle. (Clinical Cancer Research, 2011, 17(13): 4200-4207.) The cell cycle is regulated by cyclin-dependent kinases (CDKs). The CDKs family has 14 serine / threonine protein kinases. The activity of CDK is regulated by phosphorylation and binding to different cyclins. The transition of cells from the G2 phase to the division phase is positively regulated by the phosphorylation of CDK1 (also known as CDC2) and its related cyclin B. Before cell division, CDK1 is inactive, its 15th tyrosine is phosphorylated by WEE1, and then its 14th threonine is phosphorylated by myelin transcription factor (MYT1). Therefore, WEE1 is a negative regulator of the cell cycle. It negatively regulates the cell from the G2 phase to the division phase by preventing the cyclin B and activated CDK1 complex from entering the nucleus. The expression and activity of WEE1 are increased in the S phase and G2 phase, and decreased in the highly phosphorylated M phase. When cells enter the G2 phase and no DNA damage occurs, polo-like protein kinase 1 (PLK1) phosphorylates WEE1 and degrades WEE1 through the ubiquitin ligase complex. PLK1 also phosphorylates and activates protein phosphatase cell division cycle 25 analog (CDC25), which activates CDK1 by dephosphorylation. Active CDK1 can bind to cyclin B and promote cells to enter the division phase. (Molecular & Cellular Biology, 2012, 32 (20): 4226.)

[0003] When the DNA of a cell is damaged, the G1, S, and G2 checkpoints delay the cell from entering the division phase, buying time for the cell to repair the damaged DNA before entering the division phase, thereby ensuring the integrity of the genome. P53, a key regulatory factor of the G1 checkpoint, is a mutant in many malignant tumor cells. (Proceedings of the National Academy of Sciences of the United States of America, 2007, 104(10): 3753-3758.) Tumor cells with defective P53 function cannot arrest the cell cycle in the G1 phase when the DNA is damaged, and therefore rely more on the G2 checkpoint. In response to DNA damage, the G2 checkpoint inhibits the phosphorylation of CDK1 through two parallel and interconnected pathways, thereby delaying the cell from entering the division phase. Depending on the type of DNA damage, ataxia telangiectasia mutated (ATM) protein kinase or ataxia telangiectasia-related (ATR) protein kinase is activated. (Oncotarget, 2016, 7(31): 49902-49916.)

[0004] ATM is activated by ionizing radiation, radioactive agents, and agents that cause double-strand DNA breaks. ATM phosphorylates and activates checkpoint kinase 2 (CHK2), which phosphorylates Ser216 of cell division cycle 25C phosphatase (CDC25C). This leads to nuclear export and cytoplasmic separation of CDC25C, thereby inhibiting its phosphorylation activity. Inhibition of CDC25C activity leads to inhibition of phosphorylation of the CDK1 / CDK2-binding cyclin B complex, rendering CDK1 inactive and inhibiting cell entry into the division phase. (Molecular Cancer, 2014, 13(1): 72.)

[0005] ATR is activated by a wide range of genotoxic stimuli that cause single-strand DNA breaks. ATR is the main kinase responsible for phosphorylating and activating CHK1. In contrast to CHK2, which can only be activated by ATM, CHK1 can be activated by both ATM and ATR. CHK1 simultaneously phosphorylates WEE1 and CDC25C, activating WEE1 kinase activity and inhibiting CDC25C phosphatase activity. WEE1 phosphorylates CDK1-binding cyclin B, resulting in cell cycle arrest in the G2 phase, providing time for DNA repair. (Drug News & Perspectives, 2010, 23(7): 425.)

[0006] WEE1 is overexpressed in many malignant tumors, such as liver cancer, breast cancer, malignant glioma, melanoma, and adult and pediatric brain tumors. Some tumor cells have abnormal G1 checkpoints. Inhibiting WEE1 activity will lead to G2 checkpoint failure. At this time, cells with unrepaired damaged DNA will continue to divide and eventually die. (Molecular Cancer Therapeutics, 2013, 12(12): 2675-2684.) Whether through pyrimidine derivatives (PD0166285) or small interfering RNA knockdown, inhibiting WEE1 activity will make ovarian cancer, colon cancer, cervical cancer, osteosarcoma, malignant glioma and lung cancer cells more sensitive to DNA damage caused by radiation and topoisomerase inhibition. Therefore, WEE1 inhibitors have broad development space both as single drugs and in combination. (Cancer Biology & Therapy, 2010, 9(7): 523-525.)

[0007] Patents WO2007126122, WO2008133866, WO2013012681, WO2013126656, WO2014167347, WO2015092431, WO2018011569, WO2018011570, WO2018090939, WO2018133829, WO2018171633, etc. describe small molecule compounds with WEE1 kinase inhibitory activity. Currently, the most advanced compound is AZD1775, which has entered Phase II clinical trials and has shown good cancer treatment effects. However, AZD1775 also has obvious adverse reactions, and its drug activity and efficacy need to be improved. Therefore, it is urgent to provide a WEE1 inhibitor with better safety and wider application population. Summary of the invention

[0008] The object of the present invention is to provide a compound as shown in Formula I, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0009]

[0010] Among them, the R 1 Selected from -C 1～6 Alkyl, -C 2～6 Alkenyl, -C 2～6 Alkynyl, -C 0～2 Alkylene -CN, -C 0～2 Alkylene-(3- to 10-membered cycloalkyl), -C 0～2 Alkylene-(3-10 membered heterocycloalkyl); the -C 0～2 Alkylene, -C 1～6 Alkyl, -C 2～6Alkenyl, -C 2～6 Alkynyl, 3- to 10-membered cycloalkyl, and 3- to 10-membered heterocycloalkyl may be further substituted by one, two, three, or four independent R 11 replace;

[0011] The R 11 Selected from hydrogen, C 1～6 Alkyl, halogen substituted C 1～6 Alkyl, halogen;

[0012] R 2 Selected from

[0013] The R 21 Selected from hydrogen, C 1～6 Alkyl, halogen substituted C 1～6 Alkyl, -C 0～2 Alkylene-C(O)OC 1～6 Alkyl, -C 0～2 Alkylene -COOH, -C(O)C 1～6 Alkyl, halogen;

[0014] The R 3 Selected from

[0015] X 1 Selected from chemical bonds, O, NR N1 or CR C1 R C2 ;X 2 Select from N or CR C1 ; The X 3 Selected from O, NR N1 or CR C1 R C2 ;

[0016] n is selected from 1, 2 or 3;

[0017] R N1 Selected from hydrogen, halogen, -OH, -C 1～6 Alkyl, halogen substituted C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 Alkyl), -NH 2 、-NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl), -C(O)(C1～6 alkyl);

[0018] R C1 , R C2 are independently selected from hydrogen, halogen, cyano, nitro, -OH, -C 1～6 Alkyl, halogen substituted C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 Alkyl), -NH 2 , -C 0～2 Alkylene-NH(C 1～6 Alkyl), -C 0～2 Alkylene-N(C 1～6 Alkyl)(C 1～6 alkyl);

[0019] R 4 Selected from -(5-12 membered heterocycloalkyl); the heterocycloalkyl may be further replaced by one, two, three or four independent R 41 replace;

[0020] The R 41 Selected from hydrogen, oxo, -C 1～6 Alkyl, -C 2～6 Alkenyl, halogen-substituted C 1～6 Alkyl, hydroxy substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -C 0～2 Alkylene-O(C 1～6 Alkyl), -O(halogen substituted C 1～6 Alkyl), -NH 2 、-NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-C(O)R 42 , -C 0～2 Alkylene-C(O)NR 42 R 43 , -C 0～2 Alkylene-C(O)OR 42 , -C 0～2 Alkylene-S(O)R 42 , -C 0～2 Alkylene-S(O)NR 42 R 43 , -C 0～2 Alkylene-S(O)OR 42 , -C 0～2 Alkylene-S(O) 2 R 42 , -C0～2 Alkylene-S(O) 2 NR 42 R 43 , -C 0～2 Alkylene-S(O) 2 OR 42 , -C 0～2 Alkylene-(3- to 10-membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl), -C 0～2 Alkylene-(5-10 membered aromatic ring), -C 0～2 Alkylene-(5-10 membered aromatic heterocycle); wherein the alkyl, alkylene, cycloalkyl, heterocycloalkyl, aromatic ring, aromatic heterocycle may be further replaced by one, two, three or four independent R 44 replace;

[0021] R 42 , R 43 are independently selected from hydrogen, halogen, cyano, nitro, -OH, -C 1～6 Alkyl, halogen substituted C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 Alkyl), -NH 2 、-NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3- to 10-membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl), -C 0～2 Alkylene-(5-10 membered aromatic ring), -C 0～2 Alkylene-(5-10 membered aromatic heterocycle);

[0022] R 44 Selected from hydrogen, oxo, halogen, cyano, nitro, -OH, -C 1～6 Alkyl, halogen-substituted -C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 Alkyl), -NH 2 , -C 0～2 Alkylene-NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl);

[0023] Alternatively, the R 41 With R C1 or R C2Together with the atoms directly connected to it, it forms a 3- to 10-membered carbocyclic ring or a 3- to 10-membered heterocyclic ring;

[0024] The R 5 Selected from hydrogen, oxo, halogen, cyano, nitro, -OH, -NH 2 , -C 1～6 Alkyl, halogen substituted C 1～6 Alkyl, hydroxy substituted C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 Alkyl), -C 0～2 Alkylene-NH(C 1～6 Alkyl), -OC 0～2 Alkylene-N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3- to 10-membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl), -O-(3- to 10-membered cycloalkyl), -OC 0～2 Alkylene-(3- to 10-membered heterocycloalkyl), -C 0～2 Alkylene-(5-10 membered aromatic ring), -C 0～2 Alkylene-(5-10 membered aromatic heterocycle); wherein the alkyl, alkylene, cycloalkyl, heterocycloalkyl, aromatic ring, aromatic heterocycle may be further replaced by one, two, three or four independent R 51 replace;

[0025] The R 51 Selected from hydrogen, C 1～6 Alkyl, halogen substituted C 1～6 Alkyl, oxo, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 Alkyl), -NH 2 , -C 0～2 Alkylene-NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl);

[0026] Each R 6 are independently selected from hydrogen, C 1～6 Alkyl, halogen substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6Alkyl), -NH 2 、-NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3- to 10-membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl), -C 0～2 Alkylene-(5-10 membered aromatic ring), -C 0～2 Alkylene-(5-10 membered aromatic heterocycle), -C 0～2 Alkylene-(5- to 12-membered spiro ring), -C 0～2 Alkylene-(5- to 12-membered spiro heterocycle), -C 0～2 Alkylene-(5-12 membered bridge ring), -C 0～2 Alkylene-(5- to 12-membered bridged heterocyclic ring);

[0027] Wherein, the A ring is selected from 5-12 membered spiro ring, 5-12 membered spiro heterocycle, 5-12 membered bridged ring, 5-12 membered bridged heterocycle; wherein the spiro ring, spiro heterocycle, bridged ring, bridged heterocycle can be further replaced by one, two, three or four independent R A1 replace;

[0028] Each R A1 are independently selected from hydrogen, halogen, cyano, nitro, -OH, -C 1～6 Alkyl, halogen substituted C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 Alkyl), -NH 2 、-NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3- to 10-membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl), -C 0～2 Alkylene-(5-10 membered aromatic ring), -C 0～2 Alkylene-(5-10 membered aromatic heterocycle), -C 0～2 Alkylene-(5- to 12-membered spiro ring), -C 0～2 Alkylene-(5- to 12-membered spiro heterocycle), -C 0～2 Alkylene-(5-12 membered bridge ring), -C 0～2 Alkylene-(5- to 12-membered bridged heterocyclic ring);

[0029] wherein the B ring is selected from a 3- to 10-membered carbocyclic ring, a 3- to 10-membered heterocyclic ring, a 5- to 10-membered aromatic ring, a 5- to 10-membered aromatic heterocyclic ring, a 5- to 12-membered spirocyclic ring, a 5- to 12-membered spiroheterocyclic ring, a 5- to 12-membered bridged ring, a 5- to 12-membered bridged heterocyclic ring, a 3- to 10-membered fused ring, and a 3- to 10-membered fused heterocyclic ring; wherein the carbocyclic ring, heterocyclic ring, aromatic ring, aromatic heterocyclic ring, spirocyclic ring, spiroheterocyclic ring, bridged ring, bridged heterocyclic ring, fused ring, and fused heterocyclic ring may be further replaced by one, two, three, or four independent R B1 replace;

[0030] Each R B1 are independently selected from hydrogen, -C 1～6 Alkyl, halogen substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 Alkyl), -NH 2 、-NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3- to 10-membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl), -C 0～2 Alkylene-(5-10 membered aromatic ring), -C 0～2 Alkylene-(5-10 membered aromatic heterocycle);

[0031] R 7 , R 8 are independently selected from hydrogen, halogen, cyano, nitro, -OH, -C 1～6 Alkyl, halogen substituted C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 Alkyl), -NH 2 、-NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3- to 10-membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl), -C 0～2 Alkylene-(5-10 membered aromatic ring), -C 0～2 Alkylene-(5-10 membered aromatic heterocycle);

[0032] or R 7 , R 8The atoms directly connected to it form a 3-10-membered fused ring, a 3-10-membered fused heterocycle, a 3-10-membered spirocycle, or a 3-10-membered spiroheterocycle; the fused ring, fused heterocycle, spirocycle, or spiroheterocycle may be further replaced by one, two, three, or four independent R 71 replace;

[0033] Each R 71 are independently selected from hydrogen, C 1～6 Alkyl, halogen substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 Alkyl), -NH 2 、-NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl);

[0034] Wherein, the C ring is selected from 5-12 membered spiro ring, 5-12 membered spiro heterocycle, 5-12 membered bridged ring, 5-12 membered bridged heterocycle, 3-10 membered condensed ring, 3-10 membered condensed heterocycle; wherein the spiro ring, spiro heterocycle, bridged ring, bridged heterocycle, condensed ring, condensed heterocycle can be further replaced by one, two, three or four independent R C1 replace;

[0035] Each R C1 are independently selected from hydrogen, halogen, cyano, nitro, -OH, C 1～6 Alkyl, halogen substituted C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 Alkyl), -NH 2 、-NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3- to 10-membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl), -C 0～2 Alkylene-(5-10 membered aromatic ring), -C 0～2 Alkylene-(5-10 membered aromatic heterocycle), -C 0～2 Alkylene-(5- to 12-membered spiro ring), -C 0～2 Alkylene-(5- to 12-membered spiro heterocycle), -C 0～2 Alkylene-(5-12 membered bridge ring), -C 0～2 Alkylene-(5- to 12-membered bridged heterocyclic ring);

[0036] The R 10 Selected from hydrogen, C 1～6Alkyl, halogen substituted C 1～6 alkyl;

[0037] The R 9 Selected from -C 0～2 Alkylene-C(O)NR 91 R 92 、-C(O)C 1～6 Alkyl; each R 91 , R 92 are independently selected from hydrogen, C 1～6 Alkyl, halogen substituted C 1～6 alkyl;

[0038] or R 91 , R 92 The atoms directly connected to it form a 3-10 membered heterocyclic ring, which may be further separated by one, two, three or four independent R 93 replace;

[0039] Each R 93 are independently selected from hydrogen, C 1～6 Alkyl, halogen substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 Alkyl), -NH 2 、-NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl);

[0040] When R 3 for The B ring is a 3-membered carbon ring, and R1 is When R 2 Not for

[0041] R 3 for R 7 , R 8 are hydrogen, R1 is When, R 3 Not for R 3 for X 1 is the key, R 4 for R1 is When R 2 Not for R 3 for X 1 is the key, R 4 for R1 is When R 2 Not for R 3 for R1 is When R 2 for

[0042] R3 is R1 is When R 2 Not for R 2 for R 21 is methyl, R1 is When R 3 Not for

[0043] R 2 for R 21 is hydrogen, R1 is When R 3 Not for

[0044] R 2 for R 21 is hydrogen, R1 is When R 3 Not for

[0045] R 2 for R 21 is hydrogen, R1 is When R 3 Not for

[0046] R 2 for R1 is When R 3 Not for

[0047] Preferably, the compound of the present invention, or its stereoisomer, or its pharmaceutically acceptable salt, wherein R 1 is selected from methyl, ethyl, propyl, isopropyl, cyclopropyl,

[0048] Preferably, the compound of the present invention, or its stereoisomer, or its pharmaceutically acceptable salt, wherein R 2 Selected from

[0049] Preferably, the compound of the present invention, or its stereoisomer, or its pharmaceutically acceptable salt, wherein X 1 Selected from chemical bonds, O, NR N1 or CR C1 R C2 ; R C1 , R C2 are independently selected from hydrogen, C 1～3 Alkyl, halogen, halogen-substituted C 1～3 Alkyl, -C 1～2 Alkylene-N(C 1～6 Alkyl)(C 1～6 alkyl);

[0050] R N1 Selected from hydrogen, C 1～3 Alkyl, halogen substituted C 1～3 Alkyl, -(3- to 10-membered heterocycloalkyl).

[0051] Preferably, the compound of the present invention, or its stereoisomer, or its pharmaceutically acceptable salt, wherein R 4 Selected from: Wherein, X is selected from O, NR 41 ;X 4 Selected from O, NH or CH 2 ;n 1 Selected from 1 or 2; said R 4 Can be further divided into one, two, three or four independent R 41 replace;

[0052] The R 41 Selected from hydrogen, oxo, C 1～3 Alkyl, -C 2～6 Alkenyl, halogen-substituted C 1～3 Alkyl, hydroxy substituted C 1～6 Alkyl, -C 0～2 Alkylene-O(C 1～3 Alkyl), -NH 2 、-NH(C 1～3 Alkyl), -N(C 1～3 Alkyl)(C 1～3 Alkyl), -C(O)R 42 、-C(O)NR 42 R 43 、-C(O)OR 42 , -(3- to 10-membered cycloalkyl), -(3- to 10-membered heterocycloalkyl), -C 1～2Alkylene-(5-10 membered aromatic ring); wherein the alkyl, alkylene, cycloalkyl, heterocycloalkyl, aromatic ring may be further replaced by one, two, three or four independent R 44 replace;

[0053] R 42 , R 43 are independently selected from hydrogen, halogen, -OH, C 1～3 Alkyl, halogen substituted C 1～3 alkyl;

[0054] R 44 Selected from hydrogen, C 1～3 Alkyl, halogen substituted C 1～3 Alkyl, -O(C 1～3 alkyl).

[0055] Preferably, the compound of the present invention, or its stereoisomer, or its pharmaceutically acceptable salt, wherein R 4 Specific selection:

[0056] Preferably, the compound of the present invention, or its stereoisomer, or its pharmaceutically acceptable salt, wherein R 5 Selected from hydrogen, halogen, -C 1～3 Alkyl, halogen substituted C 1～3 Alkyl, hydroxy substituted C 1～3 Alkyl, -O(C 1～3 Alkyl), -NH(C 1～3 Alkyl), -OC 2 Alkylene-N(C 1～3 Alkyl)(C 1～3 alkyl), -O-(3- to 10-membered cycloalkyl), -O-(3- to 10-membered heterocycloalkyl), wherein the alkyl, alkylene, cycloalkyl, heterocycloalkyl may be further replaced by one, two, three or four independent R 51 replace;

[0057] The R 51 Selected from hydrogen, C 1～3 Alkyl, halogen substituted C 1～3 alkyl.

[0058] Preferably, the compound of the present invention, or its stereoisomer, or its pharmaceutically acceptable salt, wherein R 5 Specifically selected from methyl, ethyl, ethoxy, F, hydroxy-substituted methyl,

[0059] Preferably, the compound of the present invention, or its stereoisomer, or its pharmaceutically acceptable salt, wherein R 6 Selected from hydrogen, C 1～3 Alkyl, -NH(C 1～3 Alkyl), -N(C 1～3 Alkyl)(C 1～3 alkyl).

[0060] Preferably, the compound of the present invention, or its stereoisomer, or its pharmaceutically acceptable salt, wherein the ring A is selected from

[0061] Preferably, the compound of the present invention, or its stereoisomer, or its pharmaceutically acceptable salt, wherein the B ring is selected from a 3-10 membered heterocycle, a 5-12 membered bridged heterocycle, or a 3-10 membered fused heterocycle; wherein the heterocycle, bridged heterocycle, or fused heterocycle may be further replaced by one, two, three, or four independent R B1 replace;

[0062] Each R B1 are independently selected from hydrogen, C 1～3 Alkyl, halogen substituted C 1～3 alkyl.

[0063] Further: the B ring is selected from

[0064] Preferably, the compound of the present invention, or its stereoisomer, or its pharmaceutically acceptable salt, wherein the C ring is selected from

[0065] Preferably, the compound of the present invention, or its stereoisomer, or its pharmaceutically acceptable salt, wherein R 7 , R 8 are independently selected from -OH, or R 7 , R 8 The atoms directly connected to it form

[0066] Preferably, the compound of the present invention, or its stereoisomer, or its pharmaceutically acceptable salt, wherein R 3 Selected from

[0067]

[0068]

[0069]

[0070] Preferably, the compound of the present invention, or its stereoisomer, or its pharmaceutically acceptable salt, the compound of formula I is specifically:

[0071]

[0072]

[0073]

[0074]

[0075]

[0076]

[0077]

[0078]

[0079]

[0080]

[0081]

[0082]

[0083]

[0084]

[0085]

[0086]

[0087]

[0088]

[0089]

[0090]

[0091]

[0092]

[0093] Preferably, the compound of the present invention, or its stereoisomer, or its pharmaceutically acceptable salt, is characterized in that: the compound described in Formula I is specifically:

[0094]

[0095]

[0096]

[0097]

[0098]

[0099]

[0100] Preferably, the compound of the present invention, or its stereoisomer, or its pharmaceutically acceptable salt, is characterized in that: the compound described in Formula I is specifically:

[0101]

[0102]

[0103] The present invention also provides a use of any of the above compounds, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a WEE1-mediated disease.

[0104] The WEE1-mediated disease is one or more of diseases associated with inflammation, autoimmune diseases, infectious diseases, cancer, and precancerous syndrome.

[0105] The present invention also provides a pharmaceutical composition, which is a preparation prepared by using any of the above-mentioned compounds, or their stereoisomers, or their pharmaceutically acceptable salts as pharmaceutical active ingredients and adding pharmaceutically acceptable excipients.

[0106] "Cancer" or "malignancy" refers to any of a variety of diseases characterized by uncontrolled abnormal proliferation of cells, the ability of affected cells to spread locally or through the bloodstream and lymphatic system to other parts of the body (i.e., metastasis), and any of a number of characteristic structural and / or molecular features. "Cancer cells" refer to cells in the early, middle, or late stages of multi-step tumor progression. Cancers include sarcomas, breast cancer, lung cancer, brain cancer, bone cancer, liver cancer, kidney cancer, colon cancer, and prostate cancer. In some embodiments, the compound of Formula I is used to treat a cancer selected from colon cancer, brain cancer, breast cancer, fibrosarcoma, and squamous cell carcinoma. In some embodiments, cancer is selected from melanoma, breast cancer, colon cancer, lung cancer, and ovarian cancer. In some embodiments, the cancer treated is a metastatic cancer.

[0107] Autoimmune diseases are caused by the body's immune response to substances and tissues that are normally present in the body. Examples of autoimmune diseases include myocarditis, lupus nephritis, primary biliary cirrhosis, psoriasis, type 1 diabetes, Grave's disease, celiac disease, Crohn's disease, autoimmune neutropenia, juvenile arthritis, rheumatoid arthritis, fibromyalgia, Guillain-Barré syndrome, multiple sclerosis, and autoimmune retinopathy. Some embodiments of the invention relate to treating autoimmune diseases such as psoriasis or multiple sclerosis.

[0108] Inflammatory diseases include a variety of conditions characterized by histopathological inflammation. Examples of inflammatory diseases include acne vulgaris, asthma, celiac disease, chronic prostatitis, glomerulonephritis, inflammatory bowel disease, pelvic inflammatory disease, reperfusion injury, rheumatoid arthritis, sarcoidosis, vasculitis, airway inflammation caused by house dust mites, and interstitial cystitis. There is a significant overlap between inflammatory diseases and autoimmune diseases. Some embodiments of the present invention relate to the treatment of the inflammatory disease asthma. The immune system is often involved in inflammatory diseases, which are manifested in allergic reactions and some myopathies, and many immune system diseases lead to abnormal inflammation. IL-17A-mediated diseases also include autoimmune inflammatory diseases.

[0109] The compounds and derivatives provided in the present invention can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature system.

[0110] Definitions of terms used in the present invention: Unless otherwise stated, the initial definitions provided for groups or terms in this document apply to the groups or terms throughout the specification; for terms that are not specifically defined in this document, the meaning that a person skilled in the art can give them should be given based on the disclosure and context.

[0111] "Substitution" refers to the replacement of a hydrogen atom in a molecule by another different atom or molecule.

[0112] The minimum and maximum carbon atom content in a hydrocarbon group is indicated by a prefix, for example, the prefix C a～b Alkyl refers to any alkyl group containing from "a" to "b" carbon atoms. Thus, for example, "C 1～4 The term "alkyl" refers to an alkyl group containing 1 to 4 carbon atoms.

[0113] "Alkyl" refers to a saturated hydrocarbon chain having the specified number of member atoms. 1 ~ 6Alkyl refers to an alkyl group having 1 to 6 member atoms, for example 1 to 4 member atoms. The alkyl group can be straight or branched. Representative branched alkyl groups have one, two or three branches. The alkyl group may be optionally substituted with one or more substituents as defined herein. Alkyl includes methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl and tert-butyl), pentyl (n-pentyl, isopentyl and neopentyl) and hexyl. The alkyl group may also be part of other groups, such as C 1 ~C 6 Alkoxy.

[0114] "Cycloalkyl", "cycloalkane" refers to a saturated or partially saturated cyclic group having carbon atoms and no ring heteroatoms and having a single ring or multiple rings (including fused, combined and). For polycyclic systems with aromatic and non-aromatic rings without ring heteroatoms, the term "cycloalkyl" is applicable when the point of attachment is at a non-aromatic carbon atom (e.g., 5,6,7,8,-tetrahydronaphthalene-5-yl). The term "cycloalkyl" includes cycloalkenyl groups such as cyclohexenyl. Examples of cycloalkyl groups include, for example, adamantyl, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cyclooctyl, cyclopentenyl and cyclohexenyl. Examples of cycloalkyl groups including polybicycloalkyl ring systems are bicyclohexyl, bicyclopentyl, bicyclooctyl, etc. For example

[0115] "Alkenyl" refers to a straight or branched chain hydrocarbon group having 2 to 10 carbon atoms, and in some embodiments 2 to 6 carbon atoms or 2 to 4 carbon atoms, and having at least 1 site of vinyl unsaturation (>C=C<). For example, (Ca-Cb)alkenyl refers to an alkenyl group having a to b carbon atoms and is meant to include, for example, ethenyl, propenyl, isopropenyl, 1,3-butadienyl, and the like.

[0116] "Alkynyl" refers to a straight chain monovalent hydrocarbon radical or a branched monovalent hydrocarbon radical containing at least one triple bond. The term "alkynyl" is also intended to include those hydrocarbon groups having one triple bond and one double bond. For example, (C2-C6)alkynyl is intended to include ethynyl, propynyl, and the like.

[0117] "Halogen" is fluorine, chlorine, bromine or iodine.

[0118] "Haloalkyl" means that the hydrogen atoms in the alkyl group may be replaced by one or more halogen atoms. 1～4 The halogenalkyl group refers to an alkyl group containing 1 to 4 carbon atoms in which hydrogen atoms are substituted by one or more halogen atoms.

[0119] "Heterocycle", "heterocycloalkyl" and "heterocycloalkane" refer to a saturated ring or a non-aromatic unsaturated ring containing at least one heteroatom; wherein the heteroatom refers to a nitrogen atom, an oxygen atom, or a sulfur atom;

[0120] "Aromatic heterocycle" refers to an aromatic unsaturated ring containing at least one heteroatom; wherein the heteroatom refers to a nitrogen atom, an oxygen atom, or a sulfur atom;

[0121] "Stereoisomers" include enantiomers and diastereomers; the compounds of the present invention may contain asymmetric centers or chiral centers and therefore exist as different stereoisomers. All stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers, atropisomers, and mixtures thereof, such as racemic mixtures, form part of the present invention. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane polarized light. When describing optically active compounds, the prefix D, L or R, S is used to indicate the absolute configuration of the chiral center of the molecule. The chemical structure of these stereoisomers is the same, but their stereostructure is different. Specific stereoisomers can be enantiomers, and mixtures of isomers are usually called enantiomeric mixtures. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which may result in no stereoselectivity or stereospecificity during chemical reactions. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomers that lacks optical activity. For example, the C atom marked with "*" on the ring of the compound No. Wee1-92 in the table of the present invention is a chiral center.

[0122]

[0123] The term "pharmaceutically acceptable" means that a carrier, vehicle, diluent, excipient, and / or formed salt is generally chemically or physically compatible with the other ingredients that constitute a pharmaceutical dosage form and physiologically compatible with the receptor.

[0124] The pharmaceutical composition of the present invention can be any reusable pharmaceutical preparation form, such as oral, injection, external use, etc., and the oral dosage form includes but is not limited to tablets, capsules, oral liquids, granules, pills, suspensions, injections selected from water injections and powder injections, and external preparations selected from patches and ointments. All preparations can be prepared according to conventional pharmaceutics techniques, such as using the compound of the present invention, or any one of its stereoisomers, or its pharmaceutically acceptable salt as a pharmaceutically active ingredient, adding a pharmaceutically acceptable carrier if necessary, to prepare the above-mentioned pharmaceutical dosage form suitable for taking, wherein the pharmaceutically active ingredient per unit dose can be 0.1 mg-1000 mg, such as each tablet containing 0.1 mg-1000 mg, preferably 5-500 mg, of the pharmaceutically active ingredient.

[0125] The terms "salt" and "pharmaceutically acceptable salt" refer to acidic and / or basic salts of the above-mentioned compounds or their stereoisomers, formed with inorganic and / or organic acids and bases, and also include zwitterionic salts (inner salts), and also include quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly in the final separation and purification of the compound. It can also be obtained by mixing the above-mentioned compound, or its stereoisomer, with a certain amount of acid or base appropriately (e.g., equivalent). These salts may form a precipitate in the solution and be collected by filtering, or be recovered after solvent evaporation, or be obtained by freeze drying after reaction in an aqueous medium. The salt described in the present invention can be a hydrochloride, sulfate, citrate, benzenesulfonate, hydrobromide, hydrofluoride, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate of the compound.

[0126] In certain embodiments, one or more compounds of the present invention may be used in combination with each other. It is also possible to select the compounds of the present invention and any other active agent for use in combination to prepare a drug or pharmaceutical composition for regulating cell function or treating a disease. If a group of compounds is used, these compounds may be administered to a subject simultaneously, separately or sequentially.

[0127] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0128] The beneficial effects of the present invention are:

[0129] Compared with the prior art, the compound of the present invention has better wee1 kinase inhibitory activity and tumor cell proliferation inhibitory activity; the inhibition of cytochrome P450 enzyme and hERG potassium ion channel is reduced, and the safety is better.

[0130] The above contents of the present invention are further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. DETAILED DESCRIPTION

[0131] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and mass spectrometry (MS). NMR shifts (δ) are given in units of 10-6 (ppm). NMR measurements were performed using a Bruker AvanceIII 400 and Bruker Avance 300 NMR spectrometer, and the measurement solvents were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl 3), deuterated methanol (CD3OD), and tetramethylsilane (TMS) as the internal standard.

[0132] LC-MS was determined using a Shimadzu LC-MS 2020 (ESI). HPLC was determined using a Shimadzu LC-20A high pressure liquid chromatograph. MPLC (medium pressure preparative chromatography) used a Gilson GX-281 reverse phase preparative chromatograph. The thin layer chromatography silica gel plate used Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, and the specifications used for thin layer chromatography separation and purification products were 0.4 mm to 0.5 mm. Column chromatography generally uses Yantai Huanghai silica gel 200 to 300 mesh silica gel as a carrier.

[0133] The known starting materials of the present invention can be synthesized by methods known in the art, or can be purchased from companies such as Anaiji Chemical, Chengdu Kelon Chemical, Shaoyuan Chemical Technology, and Bailingwei Technology.

[0134] Unless otherwise specified in the examples, the reaction is carried out under a nitrogen atmosphere. Unless otherwise specified in the examples, the solution refers to an aqueous solution. Unless otherwise specified in the examples, the reaction temperature is room temperature. Unless otherwise specified in the examples, M is moles per liter.

[0135] THF: tetrahydrofuran; DIPEA: N,N-diisopropylethylamine;

[0136] DCM: dichloromethane; TFA: trifluoroacetic acid; m-CPBA: meta-chloroperbenzoic acid;

[0137] DMF: dimethylformamide; PTSA: p-toluenesulfonamide;

[0138] DMSO: dimethyl sulfoxide; NBS: N-bromosuccinimide; AIBN: azobisisobutyronitrile;

[0139] Unless otherwise specified in the examples, the HPLC test conditions are as follows:

[0140] Method A:

[0141] Column specifications: Boston Green C18 150mm*4.6mm 5um; Mobile phase: A: 0.05% TFA Water B: 0.05% TFA Acétonitrile; Gradient: B from 5% to 95% in 10.0min and hold 95% for 5.0min; Flow rate: 1.5mL / min; Column temperature: 40°C.

[0142] Method B:

[0143] Column specification: Boston Green ODS 150mm*4.6mm 5um; Mobile phase: A: 0.01M NH 4 HCO 3 Water B: Acétonitrile; Gradient: B from 5% to 95% in 10.0 min and hold 95% for 5.0 min; Flow rate: 1.5 mL / min; Column temperature: 40°C.

[0144] Unless otherwise specified, the separation conditions of SFC are: column specifications: 3μm, 150mm*3mm, mobile phase: A: CO2, mobile phase B: ethanol; flow rate: 1mL / min, column temperature: 40℃.

[0145] Intermediate Example 1: Synthesis of Intermediate IM-1:

[0146]

[0147] Step 1: Synthesis of compound IM-1-3:

[0148] Add substrate IM-1-2 (8.15 g, 35 mmol) and THF (50 mL) to a dry single-mouth bottle, stir to dissolve, add IM-1-1 (6.3 g, 37 mol) and DIPEA (15 mL, 75 mol), heat to 110 ° C for reaction, and monitor by LC-MS. After the reaction is completed, wait for the system to return to room temperature, filter out the precipitated solid, and dry it in an oven to obtain the crude product IM-1-3 (9.87 g, 76.6% yield). LCMS (ESI + )m / z:369.2[M+H] + .

[0149] Step 2: Synthesis of compound IM-1-4:

[0150] Add substrate IM-1-3 (9.16 g, 27.5 mmol) and DCM (18 mL) to a dry single-mouth bottle, stir to dissolve, slowly add TFA (18 mL), heat to 75 ° C for reaction, and monitor by LC-MS. After the reaction is completed, the organic solvent is concentrated under reduced pressure, dissolved in ethanol, and then 6 M NaOH solution is added and stirred at room temperature, and monitored by LC-MS. After the reaction is completed, solids are precipitated after reduced pressure concentration. The solids are filtered out, washed three times with water, washed three times with cold ethanol, and dried at room temperature to obtain the crude product IM-1-4 (5 g, 81.7% yield), LCMS (ESI + )m / z:223.1[M+H] + .

[0151] Step 3: Synthesis of compound IM-1:

[0152] Add substrate IM-1-4 (5.20 g, 23.4 mmol), potassium carbonate (4.53 g, 32.8 mmol) and IM-1-5 (5.55 g, 25.7 mmol) to a dry two-necked flask. After nitrogen replacement three times, add cuprous iodide (4.46 g, 23.4 mmol), N,N'-dimethylethylenediamine (4.13 g, 46.8 mmol) and 1,4-dioxane (100 mL). After stirring evenly, heat to 120 ° C for reaction, and monitor by LC-MS and TLC. After the reaction is completed, add ammonia water and stir, extract 3 times with ethyl acetate, combine the organic phases, remove water with anhydrous sodium sulfate, and concentrate the organic phase under reduced pressure. The residue is purified by medium pressure liquid chromatography (alkali method) to obtain compound IM-1 (5.1 g, 61% yield), LCMS (ESI + )m / z:358.1[M+H] + .

[0153] Intermediate Example 2: Synthesis of Intermediate IM-2

[0154]

[0155] Step 1: Synthesis of compound IM-2-3:

[0156] Add substrate IM-2-1 (7.12 g, 50 mmol) and DMSO (30 mL) to a dry single-mouth bottle, stir and dissolve, then add IM-2-2 (9.9 g, 50 mmol), heat to 90 °C for reaction, and monitor with LC-MS. After the reaction is completed, cool to room temperature, filter out the precipitated solid, wash three times with water, wash three times with a small amount of methanol, and dry to obtain the product IM-2-3 (12.12 g, 76% yield), LCMS (ESI + )m / z:320.3[M+H] + .

[0157] Step 2: Synthesis of compound IM-2-4:

[0158] Add substrate IM-2-3 (12.12 g, 38 mmol) and DCM (18 mL) to a dry single-mouth bottle, stir to dissolve, slowly add TFA (18 mL), and monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure to remove the solvent to obtain a residue. Add methanol (70 mL) to the residue, stir to dissolve, add paraformaldehyde (7 g) and acetic acid (1 mL), stir at room temperature for 30 min, add sodium cyanoborohydride (3.8 g, 60 mmol), and heat to 50 ° C for reaction. After the reaction is completed, cool to room temperature, filter out the solid, concentrate the organic phase under reduced pressure to remove the solvent, and purify the residue by medium pressure liquid chromatography (alkaline method) to obtain compound IM-2-4 (7.6 g, 85.8% yield), LCMS (ESI + )m / z:234.2[M+H] + .

[0159] Step 3: Synthesis of compound IM-2:

[0160] Add substrate IM-2-4 (5 g, 21.5 mmol) and methanol (40 mL) to a dry single-mouth bottle, stir and dissolve, add Pd / C (500 mg), replace with hydrogen five times, stir at room temperature to react, and monitor with LC-MS. After the reaction, remove Pd / C with diatomaceous earth, and concentrate the organic phase under reduced pressure to obtain product IM-2 (4.0 g, crude), LCMS (ESI + )m / z:204.2[M+H] + .

[0161] Intermediate Example 3: Synthesis of Intermediate IM-3

[0162]

[0163] Step 1: Synthesis of compound IM-3:

[0164] Substrate IM-1-4 (138 mg, 620.88 μmol), potassium carbonate (101 mg, 620.88 μmol) and IM-3-1 (130 mg, 521.82 μmol) were added to a dry two-necked bottle. After nitrogen replacement three times, cuprous iodide (99 mg, 519.82 μmol), N, N'-dimethylethylenediamine (106 mg, 1.20 mmol) and 1,4-dioxane (10 mL) were added. After stirring evenly, the temperature was raised to 120 ° C for reaction and LC-MS monitoring was performed. After the reaction was completed, the temperature was restored to room temperature, ammonia water was added, and ethyl acetate was extracted three times. After the organic phase was concentrated under reduced pressure, the residue was purified by medium pressure liquid chromatography (alkali method) to obtain compound IM-3 (130 mg, 53.62% yield), LCMS (E+) m / z: 391.2 [M+H] + .

[0165] Intermediate Example 4: Synthesis of Intermediate IM-4

[0166]

[0167] Step 1: Synthesis of compound IM-4:

[0168] Add substrate IM-2-3 (1 g, 3.13 mmol) and methanol (10 mL) to a dry single-mouth bottle, stir and dissolve, add Pd / C (200 mg), replace with hydrogen five times, stir at room temperature to react, and monitor with LC-MS. After the reaction, remove Pd / C with diatomaceous earth, and concentrate the organic phase under reduced pressure to obtain product IM-4 (700 mg, crude), LCMS (ESI + )m / z:290.2[M+H] + .

[0169] Intermediate Example 5: Synthesis of Intermediate IM-5

[0170]

[0171] Step 1: Synthesis of compound IM-5:

[0172] Add substrate IM-3 (0.2 g, 0.51 mmol) to a single-mouth reaction bottle, stir and dissolve in anhydrous THF (5 ml), add m-CPBA (208.2 g, 1.02 mmol, purity 85%), stir at room temperature for 40 minutes, and monitor by LC-MS. The reactants were concentrated under reduced pressure and separated by medium pressure liquid chromatography to obtain IM-5 (160 mg), LCMS (ESI + )m / z:407.1[M+H] + .

[0173] Intermediate Example 6: Synthesis of Intermediate IM-6

[0174]

[0175] Step 1: Synthesis of compound IM-6:

[0176] Substrate IM-1 (0.2 g, 0.56 mmol) was added to a single-mouth reaction bottle, anhydrous THF (5 ml) was stirred and dissolved, and m-CPBA (144.9 mg, 0.84 mmol, purity 85%) was added, and stirred at room temperature for 40 minutes, and monitored by LC-MS. The reactants were concentrated under reduced pressure and separated by medium pressure liquid chromatography to obtain IM-6 (150 mg, 0.40 mmol, 71.8% yield), LCMS (ESI + )m / z:374.1[M+H]+ .

[0177] Example 1: Synthesis of Compound 1

[0178]

[0179] Step 1: Synthesis of compound 1-2

[0180] Add 2-bromo-6-fluoropyridine (3 g, 15 mmol) and tetrahydrofuran (25 mL) to a dry round-bottom flask. The temperature drops to -40°C, and NaHMDS (12.79 g, 85.23 mmol) is slowly added dropwise to the system. After stirring at -40°C for 30 minutes, dimethyl sulfoxide (3.21 g, 34.09 mmol) is added and stirring is continued at this temperature for 3 hours. The reaction is quenched with water, extracted three times with ethyl acetate, and the organic phase is dried over anhydrous sodium sulfate, filtered, and distilled under reduced pressure. Compound 1-2 (1.5 g, 6.00 mmol) is obtained by purification by MPLC, LCMS (ESI) m / z: 252.0 [M+H] + .

[0181] Step 2: Synthesis of Compound 1-3

[0182] Compound 1-2 (1.5 g, 6.00 mmol), tetrabutylammonium bromide (193.1 mg, 0.6 mmol), and dichloromethane (10 mL) were added to a dry flask at room temperature. After stirring at room temperature for 10 minutes, sodium hydroxide (9.60 g, 239.9 mmol) and water (10 mL) were added. The reaction was stirred at room temperature for 12 hours, quenched with water and extracted with ethyl acetate three times. The combined organic phase was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by MPLC to give compound 1-3 (450 mg, 1.63 mmol), LCMS (ESI) m / z: 278.0 [M+H] + .

[0183] Step 3: Synthesis of Compound 1-4

[0184] In a dry microwave tube, under nitrogen protection, add substrate 1-3 (93 mg, 336.77 μmol), IM-1-4 (89.82 mg, 404.13 μmol), cuprous iodide (64.14 mg, 336.77 μmol), potassium carbonate (65.17 mg, 505.16 μmol) and N, N'-dimethylethylenediamine (29.64 mg, 336.77 μmol), and then add 1,4-dioxane (5 mL). Under nitrogen environment, heat the reaction at 120 ° C for 10 hours. LC-MS monitoring, after the reaction is completed, the reaction system is cooled to room temperature, filtered, extracted with ethyl acetate 3 times, the organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by MPLC to obtain compound 1-4 (50 mg, 119.76 μmol), LCMS (ESI) m / z: 418.1 [M+H] + .

[0185] Step 4: Synthesis of compound 1

[0186] To tetrahydrofuran (1 mL) containing 1-4 (30 mg, 71.9 μmol) was added m-chloroperbenzoic acid (18.74 mg, 107.8 μmol) and stirred at room temperature for 1 hour. After the disappearance of the starting material, N,N-diisopropylethylamine (43 mg, 0.33 mmol) and 1-5 (15.3 mg, 80.1 μmol) were added and stirred at room temperature for 12 hours. After the reaction was completed, the mixture was concentrated under reduced pressure and the residue was purified by preparative high performance liquid chromatography (acidic) to obtain compound 1 (30 mg, 53.51 μmol). LCMS (ESI) m / z: 561.3 [M+H] + ,HPLC method A:R T =5.05min,purity:98.2%. 1 H NMR (400 MHz, Methanol-d 6 )δ8.79(s,1H),8.05–8.01(m,1H),7.93(d,J=8.4Hz,1H),7.61(d,J=7.6Hz, 1H),7.57-7.42(m,2H),7.03-6.88(m,2H),5.71–5.67(m,1H),5.06–5.05(m ,1H),4.98–4.95(m,1H),4.80–4.78(m,2H),3.20–3.18(m,5H),3.04(s,4H) ,2.65–2.63(m,5H),2.36(s,4H),1.92-1.75(m,2H),1.53(d,J=2.4Hz,2H).

[0187] Example 2: Synthesis of Compound 2

[0188]

[0189] Step 1: Synthesis of compound 2-1:

[0190] Add substrate IM-1 (43 mg, 120.30 μmol) and toluene (1 mL) to a dry single-mouth bottle, stir and dissolve, then add m-CPBA (43.60 mg, 252.64 μmol), react at room temperature for 30 min, and monitor by LC-MS. After the reaction is completed, add DIPEA (80.6 mg, 623.65 μmol, 108.63 μL), stir for 10 min, then add IM-4 (45.14 mg, 0.156 mmol), and monitor by LC-MS. After the reaction is completed, the reaction solution is concentrated under reduced pressure, and the residue is purified by medium pressure liquid chromatography (alkaline method) to obtain compound 2-1 (45 mg, 62.64% yield). 1 H NMR (400 MHz, DMSO-d 6 )δ10.08(s,1H),8.80(s,1H),8.00(t,J=8.0Hz,1H),7.74(d,J=8.2Hz,1H),7.59(d,J=7.7Hz,1H),7. 51(s,2H),6.61(d,J=8.5Hz,2H),5.71-5.61(m,1H),5.31(s,1H),4.99(dd,J=10.2,1.5Hz,1H),4.83( dd,J=17.1,1.5Hz,1H),4.68(d,J=5.9Hz,2H),4.49(s,1H),4.42(d,J=23.5Hz,1H),3.55(t,J=10.6H z,1H),3.27–3.20(m,2H),2.96(t,J=9.0Hz,1H),1.93(s,2H),1.46(s,6H),1.39(s,5H),1.32(s,4H). 1 H NMR (400 MHz, DMSO-d 6 ,D 2O)δ8.81(s,1H),8.02(t,J=7.9Hz,1H),7.73(d,J=7.8Hz,1H),7.60(d,J=7.7Hz,1H),7.51( s,2H),6.62(d,J=8.5Hz,2H),5.71–5.61(m,1H),5.04–5.01(m,1H),4.85–4.81(m,1H),4.6 9(d,J=5.9Hz,2H),4.49–4.41(m,2H),3.57(t,J=9.0Hz,1H),3.36–3.25(m,2H),2.96(t,J= 8.4Hz,1H),1.94(s,2H),1.47(s,6H),1.39(s,5H),1.32(s,4H).LCMS(E+)m / z:599.4[M+H] + ,HPLC method B:R T =8.23min,purity:91.5%.

[0191] Step 2: Synthesis of compound 2:

[0192] Substrate 2-1 (40.00 mg, 66.81 μmol) and DCM (3 mL) were added to a dry single-mouth bottle, stirred and dissolved, and TFA (3 mL) was slowly added, and LC-MS was monitored. After the reaction was completed, the solvent was removed by concentration under reduced pressure to obtain a residue, which was purified by medium pressure liquid chromatography (acid method) to obtain compound 2 (25 mg, 61.08% yield). 1 H NMR (400 MHz, Methanol-d 4 )δ8.77(s,1H),7.97(t,J=7.9Hz,1H),7.76(d,J=8.1Hz,1H),7.63(dd,J=7.8,0.8Hz,1H),7.54( d,J=8.4Hz,2H),6.67(d,J=8.6Hz,2H),5.70(m,1H),5.03(dq,J=10.2,1.2Hz,1H),4.92(d,J=1.2 Hz,1H),4.80(d,J=6.1Hz,2H),4.66(s,1H),4.48(s,1H),3.76(dd,J=10.6,2.5Hz,1H),3.42–3.3 4(m,3H),2.32(d,J=11.1Hz,1H),2.05(d,J=11.3Hz,1H),1.57(s,6H).LCMS(E+)m / z:499.2[M+H] + ,HPLC method A:R T =5.03min,purity:99.2%.

[0193] Example 3: Synthesis of Compound 3

[0194]

[0195] Step 1: Synthesis of compound 3:

[0196] Add substrate IM-1 (3.57 g, 10 mmol) and tetrahydrofuran (30 mL) to a dry single-mouth bottle, stir and dissolve, then add m-CPBA (2.5 g, 15 mmol, purity 85%), react at room temperature for 1 h, and monitor by LC-MS. After the reaction is completed, add DIPEA (6.45 g, 50 mmol), stir for 10 min, then add IM-2 (2.1 g, 10 mmol), and monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure to remove the solvent, and the residual impurities are purified by medium pressure liquid chromatography (alkaline method) to obtain compound 3 (2.1 g, 41% yield). 1 H NMR (400 MHz, DMSO-d 6 )δ10.04(s,1H),8.79(s,1H),8.00(t,J=7.9Hz,1H),7.75(s,1H),7.59(d,J=7.7Hz,1H),7.48(s,2 H),6.56(d,J=8.5Hz,2H),5.71-5.61(m,1H),5.32(s,1H),4.99(dd,J=10.3,1.5Hz,1H),4.82(d,J= 17.1,1H),4.67(d,J=6.0Hz,2H),4.27(s,1H),3.42(s,1H),3.13(d,J=9.0Hz,1H),2.78(d,J=9.5, 1H), 2.52 (d, J = 9.7Hz, 2H), 2.25 (s, 3H), 1.87 (d, J = 9.3Hz, 1H), 1.76 (d, J = 9.2Hz, 1H), 1.46 (s, 6H). 1 H NMR (400 MHz, DMSO-d 6 ,D 2O)δ8.80(s,1H),8.02(t,J=7.9Hz,1H),7.73(s,1H),7.61(d,J=7.8Hz,1H),7.48(s,2H),6.58(d,J=8.6H z,2H),5.71-5.60(d,J=10.3Hz,1H),5.03(m,1H),4.83(d,J=17.1Hz,1H),4.68(d,J=5.9Hz,2H),4.29(s ,1H),3.49(s,1H),3.35(d,J=9.5Hz,1H),3.17(d,J=9.4Hz,1H),2.82(d,J=9.7Hz,1H),2.56(d,J=9.7Hz ,1H),2.27(s,3H),1.91(d,J=9.6Hz,1H),1.81(d,J=9.6Hz,1H),1.47(s,6H).LCMS(E+)m / z:513.2[M+H] + ,HPLC methodB:R T =6.02min,purity:98.4%.

[0197] Example 4: Synthesis of Compound 4

[0198]

[0199] Step 1: Synthesis of compound 4-3:

[0200] Add substrate 4-1 (114 mg, 910.78 μmol) and methanol (2 mL) to a dry single-necked bottle, stir to dissolve, and then add NaBH 4 (38 mg, 1.00 mmol), react at room temperature for 1 h, and monitor by LC-MS. After the reaction is completed, dilute hydrochloric acid is slowly added under ice bath to quench the reaction, and the solvent is removed by concentration under reduced pressure. DMSO (4 mL) is added to the residue, and potassium tert-butoxide (337 mg, 2.36 mmol) is added after stirring and dissolving. Compound 4-2 (141 mg, 999.29 μmol) is added, and the temperature is raised to 80 ° C for reaction, and LC-MS is monitored. After the reaction is completed, it is extracted three times with ethyl acetate, and the organic phase is concentrated under reduced pressure and purified by medium pressure liquid chromatography (alkali method) to obtain compound 4-3 (99 mg, 50.71% yield), LCMS (ESI + )m / z:249.2[M+H] + .

[0201] Step 2: Synthesis of compound 4-4:

[0202] Substrate 4-3 (99 mg, 398.75 μmol) and ethanol (4 mL) were added to a dry single-mouth bottle. After stirring to dissolve, Fe (67 mg, 1.20 mmol) and CH 3 COOH (47.89 mg, 797.50 μmol, 45.65 μL), heated to 80 ° C for reaction, LC-MS monitoring. After the reaction was completed, it was filtered with diatomaceous earth, washed with ethyl acetate, and the organic phase was concentrated under reduced pressure to obtain a crude product 4-4 (80 mg, crude), LCMS (ESI + )m / z:219.2[M+H] + .

[0203] Step 3: Synthesis of compound 4:

[0204] Add substrate IM-1 (71 mg, 0.2 mmol) and toluene (2 mL) to a dry single-mouth bottle, stir and dissolve, then add m-CPBA (52 mg, 0.3 mmol, purity 85%), react at room temperature for 1 h, and monitor by LC-MS. After the reaction is completed, add DIPEA (83.58 mg, 646.69 μmol, 112.64 μL), stir for 10 min, and then add compound 4-4 (52.39 mg, 0.24 mmol), and monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure to remove the solvent, and the residue is purified by medium pressure liquid chromatography (alkaline method) to obtain compound 4 (8 mg, 6.32% yield). 1 H NMR (400 MHz, DMSO-d 6 )δ10.18(s,1H),8.84(s,1H),8.04(t,J=7.9Hz,1H),7.73(d,J=7.2,1H),7.62-7.60(m,3H),6 .90(d,J=7.7Hz,2H),5.71-5.62(m,1H),5.33(s,1H),4.99(dd,J=10.3,1.4Hz,1H),4.82(dd,J =10.3,1.4Hz,1H),4.68(d,J=5.9Hz,2H),4.44-4.42(m,1H),2.77-2.60(m,6H),2.04-2.03(m, 1H),1.84-1.81(m,1H),1.66–1.63(m,1H),1.57-1.54(m,1H),1.46(s,6H),1.36–1.33(m,1H). 1 H NMR (400 MHz, DMSO-d 6 ,D 2O)δ8.85(s,1H),8.04(t,J=7.9Hz,1H),7.72(d,J=7.6Hz,1H),7.64-7.59(m,3H),6.92( d,J=8.5Hz,2H),5.72-5.62(m,1H),5.03(d,J=10.3Hz,1H),4.83(d,J=17.1Hz,1H),4.69 (d,J=5.9Hz,2H),4.49(s,1H),2.81–2.63(m,6H),2.06(s,1H),1.86(s,1H),1.70-1.68( m,1H),1.62(d,J=7.4Hz,1H),1.47(s,6H),1.42-1.37(m,1H).LCMS(E+)m / z:528.2[M+H] + ,HPLC method B:R T =6.34min,purity:82.9%.

[0205] Example 5: Synthesis of Compound 5

[0206]

[0207] Step 1: Synthesis of compound 5:

[0208] Add substrate IM-3 (30 mg, 77 μmol) and tetrahydrofuran (2 mL) to a dry single-mouth bottle, stir to dissolve, add m-CPBA (20 mg, 116 μmol), react at room temperature for 1 h, and monitor by LC-MS. After the reaction is completed, add DIPEA (50 mg, 385 μmol), stir for 10 min, add compound IM-2 (20 mg, 98.39 μmol), and monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure to obtain the residue, which is purified by medium pressure liquid chromatography (alkaline method) to obtain compound 5 (15 mg, 55.87% yield). 1 H NMR (400 MHz, DMSO-d 6)δ10.01(s,1H),8.77(s,1H),7.76(t,J=8.0Hz,1H),7.50(s,2H),7.30(d,J=7.9Hz,1H), 6.59-6.55(m,3H),5.68-5.58(m,1H),5.00(d,J=10.1Hz,1H),4.89(d,J=17.0Hz,1H),4. 74(s,2H),4.26(s,1H),3.40(s,1H),3.37(s,6H),3.30(s,1H),3.13(d,J=9.0Hz,1H),2. 78-2.76(m,1H),2.47(s,1H),2.24(s,3H),1.85(d,J=9.3Hz,1H),1.75(d,J=9.1Hz,1H). 1 H NMR (400 MHz, DMSO-d 6 ,D 2 O)δ8.79(s,1H),7.80(t,J=7.9Hz,1H),7.50-7.48(m,2H),7.30(d,J=7.8Hz,1H),6.64(d,J=8.0Hz,1H ),6.58(d,J=8.7Hz,2H),5.69–5.59(m,1H),5.05–5.03(m,1H),4.92–4.87(m,1H),4.74(s,2H),4.27(s ,1H),3.44(s,1H),3.36(s,6H),3.35–3.32(m,1H),3.16(d,J=9.4Hz,1H),2.79(d,J=9.7Hz,1H),2.48 (d,J=9.9Hz,1H),2.24(s,3H),1.89(d,J=9.5Hz,1H),1.78(d,J=9.4Hz,1H).LCMS(E+)m / z:546.3[M+H] + ,HPLC method B:R T =5.56min,purity:92%.

[0209] Example 6: Synthesis of Compound 6

[0210]

[0211] Step 1: Synthesis of compound 6-1:

[0212] Add substrate 4-1 (1 g, 6.2 mmol) and ethanol / water (10:1, 20 mL) to a dry single-mouth bottle, stir and dissolve, then add p-nitrobenzaldehyde (1 g, 6.6 mmol) and sodium hydroxide (620 mg, 15.5 mmol), react at room temperature, and monitor with LC-MS. After the reaction is completed, filter out the precipitated solid, wash three times with a small amount of ethanol, and dry naturally to obtain compound 6-1 (1 g, 63% yield), LCMS (E+) m / z: 259.2 [M+H] + .

[0213] Step 2: Synthesis of compound 6-2:

[0214] Substrate 6-1 (103 mg, 0.4 mmol) and methanol (5 mL) were added to a dry single-mouth bottle, stirred and dissolved, and Pd / C (15 mg) was added. The hydrogen was replaced five times, and the reaction was stirred at room temperature and monitored by LC-MS. After the reaction was completed, Pd / C was removed by diatomaceous earth filtration, and the organic phase was concentrated under reduced pressure to obtain a crude product 6-2 (80 mg, crude), LCMS (ESI + )m / z:231.2[M+H] + Step 3: Synthesis of compound 6:

[0215] Add substrate IM-3 (30 mg, 77 μmol) and tetrahydrofuran (2 mL) to a dry single-mouth bottle, stir to dissolve, add m-CPBA (20 mg, 116 μmol), react at room temperature for 1 h, and monitor by LC-MS. After the reaction is completed, add DIPEA (50 mg, 385 μmol), stir for 10 min, add compound 6-2 (20 mg, 86.84 μmol), and monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure to obtain the residue, which is purified by medium pressure liquid chromatography (alkaline method) to obtain compound 6 (5 mg, 17.74% yield). 1 H NMR (400 MHz, DMSO-d 6 )δ10.18(s,1H),8.85(s,1H),7.78(t,J=7.9Hz,1H),7.61(d,J=8.1Hz,2H),7.30 (d,J=7.9Hz,1H),7.20(d,J=8.4Hz,2H),6.61(d,J=7.9Hz,1H),5.69–5.60(m,1H) ,5.02-4.99(m,1H),4.90(dd,J=17.1,1.7Hz,1H),4.74(d,J=6.0Hz,2H),3.37(s, 6H),2.99–2.86(m,4H),2.76–2.69(m,3H),2.33–2.29(m,2H),1.92–1.88(m,3H).1 H NMR (400 MHz, DMSO-d 6 ,D 2 O)δ8.86(s,1H),7.83(t,J=7.9Hz,1H),7.60(d,J=8.0Hz,2H),7.31(d,J=7.8Hz,1H ),7.22(d,J=8.2Hz,2H),6.68(d,J=8.0Hz,1H),5.70–5.61(m,1H),5.07–5.04(m,1 H),4.90(d,J=17.2Hz,1H),4.75(d,J=5.9Hz,2H),3.36(s,6H),3.09–2.85(m,4H), 2.79–2.72(m,3H),2.38–2.32(m,2H),1.91-1.89(m,3H).LCMS(E+)m / z:573.6[M+H] + ,HPLC method B:R T =5.05min,purity:87.7%.

[0216] Example 7: Synthesis of Compound 7

[0217]

[0218] Step 1: Synthesis of compound 7:

[0219] Add substrate IM-3 (30 mg, 77 μmol) and tetrahydrofuran (2 mL) to a dry single-mouth bottle, stir and dissolve, then add m-CPBA (20 mg, 116 μmol), react at room temperature for 1 h, and monitor by LC-MS. After the reaction is completed, add DIPEA (50 mg, 385 μmol), stir for 10 min, and then add compound 4-4 (20 mg, 91.62 μmol), and monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure to remove the solvent, and purify the residue by medium pressure liquid chromatography (alkaline method) to obtain compound 7 (5 mg, 19.08% yield). 1 H NMR (400 MHz, DMSO-d 6)δ10.16(s,1H),δ8.82(s,1H),7.79(t,J=8.0Hz,1H),7.61(s,2H),7.28(d,J=7.8Hz,1H),6.8 9(d,J=9.0Hz,2H),6.60(d,J=8.1Hz,1H),5.69–5.59(m,1H),5.02–4.99(m,1H),4.89(d,J=17 .2Hz,1H),4.74(s,2H),4.41(d,J=7.8Hz,1H),3.18(s,3H),3.16(s,3H),2.79-2.70(m,6H),2 .03-2.02(m,1H),1.91-1.80(m,1H),1.67-1.63(m,1H),1.57-1.54(m,1H),1.38-1.33(m,1H). 1 H NMR (400 MHz, DMSO-d 6 ,D 2 O)δ8.83(s,1H),7.82(t,J=8.0Hz,1H),7.60(s,2H),7.29(d,J=7.8Hz,1H),6.92(d, J=8.8Hz,2H),6.66(d,J=8.0Hz,1H),5.68-5.62(m,1H),5.05(d,J=10.4Hz,1H),4.89 (d,J=16.9Hz,1H),4.75(s,2H),4.48(s,1H),3.36(s,6H),2.78-2.72(m,6H),2.05( s,1H),1.84(s,1H),1.70-1.59(m,2H),1.40-1.33(m,1H).LCMS(E+)m / z:561.5[M+H] + ,HPLC method B:R T =5.10min,purity:90.2%.

[0220] Example 8: Synthesis of Compound 8

[0221]

[0222] Step 1: Synthesis of compound 8-2:

[0223] Add substrate 4-2 (1 g, 7.09 mmol), compound 8-1 (1.70 g, 8.50 mmol) and potassium carbonate (2.45 g, 17.72 mmol) to a dry single-mouth bottle, dissolve with DMF (10 mL), heat to 90 ° C, react for 1 h, and monitor with LC-MS. After the reaction is completed, extract with water and ethyl acetate three times, combine the organic phases, wash with saturated brine, dry with anhydrous sodium sulfate, and concentrate the organic solvent under reduced pressure to obtain a crude product. No purification is required. The product can be directly used in the next step to obtain compound 8-2 (2.1 g, crude) as a yellow oil. LCMS (E+) m / z: 322.4 [M+H] + .

[0224] Step 2: Synthesis of compound 8-3:

[0225] Add substrate 8-2 (2.1 g, 6.53 mmol) to a dry single-mouth bottle and dissolve it with HCl / 1,4-dioxane (10 mL). React at room temperature for 1 h and monitor with LC-MS. After the reaction is completed, concentrate under reduced pressure, extract with saturated sodium carbonate solution and ethyl acetate three times, combine the organic phases, wash with saturated brine, dry with anhydrous sodium sulfate, and concentrate the organic solvent under reduced pressure to obtain a crude product. No purification is required. The crude product can be directly used in the next step to obtain compound 8-3 (1.3 g, crude) as a yellow oil. LCMS (E+) m / z: 222.4 [M+H] + .

[0226] Step 3: Synthesis of compound 8-4:

[0227] Substrate 8-3 (700 mg, 3.16 mmol) was added to a dry single-mouth bottle, and compound 4-1 (396.00 mg, 3.16 mmol) was dissolved in MeOH / AcOH (10 mL), and reacted at room temperature for 1 h, and monitored by LC-MS. After the reaction was completed, it was concentrated under reduced pressure, extracted with water and ethyl acetate three times, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and the organic solvent was concentrated under reduced pressure. The residual impurities were removed by MPLC to obtain compound 8-4 (300 mg, 907.93 μmol, 28.70% yield) as a yellow solid. LCMS (E+) m / z: 331.4 [M+H] + .

[0228] Step 4: Synthesis of compound 8-5:

[0229] Substrate 8-4 (300 mg, 907.93 μmol) was added to a dry single-mouth bottle, and formaldehyde (136.31 mg, 4.54 mmol) was dissolved in MeOH / AcOH (8 mL). The temperature was raised to 50 °C, and after reacting for 3 h, NaCNBH was added to the system. 3(142.64 mg, 2.27 mmol) was heated to 70 ° C, reacted overnight, and monitored by LC-MS. After the reaction was completed, it was concentrated under reduced pressure, extracted with water and ethyl acetate three times, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and the organic solvent was concentrated under reduced pressure. The residual impurities were removed by MPLC to obtain compound 8-5 (283 mg, 739.44 μmol, 81.44% yield) as a yellow solid. LCMS (E+) m / z: 345.5 [M+H] + .

[0230] Step 5: Synthesis of compound 8-6:

[0231] Substrate 8-5 (1.5 g, 4.74 mmol) was added to a dry single-necked bottle and dissolved in MeOH / AcOH (10 mL), and Pd / C (172.73 mg, 1.42 mmol) was added. 2 Replace 3 times, react at room temperature for 3 hours, and monitor with LC-MS. After the reaction is completed, filter with diatomaceous earth, and concentrate the filtrate under reduced pressure without purification. The crude product can be directly used in the next step to obtain compound 8-6 (1.5 g, crude) as a black oil. LCMS (E+) m / z: 315.5 [M+H] + .

[0232] Step 6: Synthesis of compound 8:

[0233] Substrate 8-6 (50 mg, 159.00 μmol) was added to a dry single-mouth bottle, compound IM--5 (64.63 mg, 159.00 μmol) was dissolved in THF (3 mL), and PTSA (54.76 mg, 318.00 μmol) was added, the temperature was raised to 110°C, and the reaction was stirred for 5 hours, and LC-MS was monitored. After the reaction was completed, the organic solvent was concentrated under reduced pressure, extracted with water and ethyl acetate three times, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residual impurities were removed by MPLC to obtain compound 8 (23 mg, 35.02 μmol, 22.02% yield) as a yellow solid. 1 H NMR (600 MHz, DMSO-d 6)δ10.11(s,1H),8.80(s,1H),7.79(s,1H),7.56(s,2H),7.30(d,J=7.9Hz,1H),6.91(d,J=8.6Hz,2H) ,6.59(d,J=8.2Hz,1H),5.68–5.59(m,1H),5.00(d,J=10.1Hz,1H),4.89(d,J=17.0Hz,1H),4.74(s,2 H),3.74–3.66(m,2H),3.37(s,6H),2.94–2.91(m,1H),2.79–2.57(m,9H),2.06(s,3H),1.97–1.93(m ,1H),1.75–1.53(m,6H),1.42(d,J=11.1Hz,1H),1.24(d,J=11.7Hz,1H).LCMS(ESI+)m / z:657.4[M+H] + ,HPLC method B:R T =5.54min,purity:98.9%.

[0234] Example 9: Synthesis of Compound 9

[0235]

[0236] Step 1: Synthesis of compound 9-2:

[0237] Substrate 4-1 (161 mg, 1 mmol) was added to a dry single-mouth bottle, and after stirring and dissolving with ethanol / water (10:1, 10 mL), 9-1 (186 mg, 1.1 mmol) and sodium hydroxide (100 mg, 2.5 mmol) were added, and the reaction was carried out at room temperature and monitored by LC-MS. After the reaction was completed, the precipitated solid was filtered out, washed three times with a small amount of ethanol, and dried to obtain compound 9-2 (150 mg, 50% yield), LCMS (E+) m / z: 303.1 [M+H] + .

[0238] Step 2: Synthesis of compound 9-3:

[0239] Substrate 9-2 (81 mg, 0.27 mmol) and methanol (5 mL) were added to a dry single-mouth bottle, stirred and dissolved, and Pd / C (15 mg) was added. The hydrogen gas was replaced five times, and the reaction was stirred at room temperature and monitored by LC-MS. After the reaction was completed, Pd / C was removed with diatomaceous earth, and the organic phase was concentrated by vacuum distillation to obtain product 9-3 (42 mg, crude), LCMS (ESI + )m / z:275.2[M+H] + .

[0240] Step 3: Synthesis of compound 9:

[0241] Add substrate IM-3 (40 mg, 110.71 μmol) and THF (2 mL) to a dry single-mouth bottle, stir and dissolve, then add m-CPBA (29 mg, 116 μmol), react at room temperature for 1 h, and monitor by LC-MS. After the reaction is completed, add DIPEA (75.12 mg, 581.26 μmol), stir for 10 min, and then add compound 9-3 (36.45 mg, 132.85 μmol), and monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure to obtain the residue, which is purified by medium pressure liquid chromatography (alkaline method) to obtain compound 9 (35 mg, 51.26% yield). 1 H NMR (400 MHz, DMSO-d 6 )δ10.12(s,1H),8.82(s,1H),7.77-7.75(m,1H),7.59(s,1H),7.45(d,J=8.9Hz,1H),7.29(d,J =7.4Hz,1H),6.89(d,J=8.8Hz,1H),6.58(d,J=8.0Hz,1H),5.67-5.60(m,1H),5.00(d,J=10.2Hz ,1H),4.88(d,J=17.1Hz,1H),4.74(s,2H),4.01(q,J=7.0Hz,2H),3.37(s,6H),3.06-3.03(m,2H ),2.85-2.84(m,1H),2.71-2.64(m,4H),2.28(s,1H),1.95–1.87(m,4H),1.32(t,J=6.9Hz,3H). 1 H NMR (400 MHz, DMSO-d 6 ,D 2O)δ8.82(s,1H),7.81-7.77(m,1H),7.58(s,1H),7.45(d,J=8.7Hz,1H),7.29(d,J=7.9Hz,1H),6.9 0(d,J=8.9Hz,1H),6.61(d,J=8.1Hz,1H),5.67-5.60(m,1H),5.02(d,J=10.2Hz,1H),4.88(d,J=17 .1Hz,1H),4.73(s,1H),4.01(q,J=7.0Hz,2H),3.36(s,6H),3.03(d,J=14.9Hz,2H),2.86(s,1H),2 .71-2.65(m,4H),2.29(s,1H),1.96-1.87(m,4H),1.32(t,J=6.9Hz,3H).LCMS(E+)m / z:617.2[M+H] + ,HPLC method B:R T =6.66min,purity:95.4%.

[0242] Example 10: Synthesis of Compound 10

[0243]

[0244] Step 1: Synthesis of compound 10-1:

[0245] Substrate 4-1 (1 g, 6.2 mmol) and ethanol / water (10:1, 20 mL) were added to a dry single-mouth bottle. After stirring and dissolving, m-nitrobenzaldehyde (1 g, 6.6 mmol) and sodium hydroxide (620 mg, 15.5 mmol) were added. The reaction was carried out at room temperature and monitored by LC-MS. After the reaction was completed, the precipitated solid was filtered out, washed three times with a small amount of ethanol, and dried naturally to obtain compound 10-1 (1 g, 63% yield). LCMS (E+) m / z: 259.2 [M+H] + .

[0246] Step 2: Synthesis of compound 10-2:

[0247] Substrate 10-1 (103 mg, 0.4 mmol) and methanol (5 mL) were added to a dry single-mouth bottle, stirred and dissolved, and Pd / C (15 mg) was added. The hydrogen gas was replaced five times, and the reaction was stirred at room temperature and monitored by LC-MS. After the reaction was completed, Pd / C was removed with diatomaceous earth, and the organic phase was concentrated by vacuum distillation to obtain product 10-2 (65 mg, crude), LCMS (ESI + )m / z:231.2[M+H] + .

[0248] Step 3: Synthesis of compound 10:

[0249] Add substrate IM-3 (40 mg, 110.71 μmol) and THF (2 mL) to a dry single-mouth bottle, stir and dissolve, then add m-CPBA (29 mg, 116 μmol), react at room temperature for 1 h, and monitor by LC-MS. After the reaction is completed, add DIPEA (85.85 mg, 664.24 μmol), stir for 10 min, and then add compound 10-2 (30.6 mg, 132.85 μmol), and monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure, and the residue is purified by medium pressure liquid chromatography (alkaline method) to obtain compound 10 (15 mg, 22.22% yield). 1 HNMR (400 MHz, DMSO-d 6 )δ10.22(s,1H),8.87(s,1H),7.83(t,J=7.9Hz,1H),7.76(s,1H),7.47(d,J=8.2Hz,1H),7.31 (d,J=7.8Hz,1H),7.21(t,J=7.8Hz,1H),6.95(d,J=7.6Hz,1H),6.61(d,J=8.1Hz,1H),5.69-5. 60(m,1H),5.01(d,J=10.1Hz,1H),4.89(d,J=17.0Hz,1H),4.74(d,J=6.0Hz,2H),3.41(s,6H)3 .03–2.98(m,2H),2.89-2.86(m,1H),2.78–2.71(m,4H),2.32-2.31(m,1H),1.97-1.89(m,4H). 1 H NMR (400 MHz, DMSO-d 6 ,D 2O)δ8.87(s,1H),7.84(t,J=7.9Hz,1H),7.74(s,1H),7.47(d,J=8.2Hz,1H),7.31(d,J=7.7Hz,1H),7 .23(t,J=7.8Hz,1H),6.96(d,J=7.7Hz,1H),6.64(d,J=8.0Hz,1H),5.70-5.60(m,1H),5.03(d,J=10. 2Hz,1H),4.89(d,J=17.0Hz,1H),4.74(d,J=6.0Hz,2H),3.36(d,J=2.1Hz,6H),3.03–2.98(m,2H),2. 93-2.86(m,1H),2.78–2.69(m,4H),2.33-2.32(m,1H),1.99-1.84(m,4H).LCMS(E+)m / z:573.1[M+H] + ,HPLC method B:R T =6.24min,purity:93.9%.

[0250] Example 11: Synthesis of Compound 11

[0251]

[0252] Step 1: Synthesis of compound 11-2:

[0253] Add substrate 4-2 (2 g, 14.17 mmol), compound 11-1 (3.17 g, 17.01 mmol) and potassium carbonate (4.89 g, 35.44 mmol) to a dry single-mouth bottle, dissolve with DMF (10 mL), heat to 90 ° C, react for 1 h, and monitor with LC-MS. After the reaction is completed, extract with water and ethyl acetate three times, combine the organic phases, wash with saturated brine, dry with anhydrous sodium sulfate, and concentrate the organic solvent under reduced pressure to obtain a crude product without purification. The product can be directly used in the next step to obtain compound 11-2 (4 g, crude) as a yellow solid. LCMS (E+) m / z: 308.3 [M+H] + .

[0254] Step 2: Synthesis of compound 11-3:

[0255] Add substrate 11-2 (4 g, 13.01 mmol) to a dry single-mouth bottle and dissolve it with HCl / 1,4-dioxane (10 mL). React at room temperature for 1 h and monitor with LC-MS. After the reaction is completed, the organic phase is concentrated under reduced pressure, extracted with saturated sodium carbonate and ethyl acetate three times, the organic phases are combined, washed with saturated brine, dried over anhydrous sodium sulfate, and the organic solvent is concentrated under reduced pressure to obtain a crude product. No purification is required, and the product can be directly used in the next step to obtain compound 11-3 (2.5 g, crude) as a yellow solid. LCMS (E+) m / z: 208.2 [M+H] + .

[0256] Step 3: Synthesis of compound 11-4:

[0257] Substrate 11-3 (1 g, 4.83 mmol) was added to a dry single-mouth bottle, and the free base of compound 4-1 (724.81 mg, 5.79 mmol) was dissolved in MeOH / AcOH (10 mL), and the reaction was carried out at room temperature for 1 h, and LC-MS was monitored. After the reaction was completed, the organic phase was concentrated under reduced pressure, extracted with saturated sodium carbonate and ethyl acetate three times, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and the organic solvent was concentrated under reduced pressure. The residual impurities were removed by MPLC to obtain compound 11-4 (1.5 g, 4.27 mmol, 88.42% yield) as a yellow solid. LCMS (E+) m / z: 317.4 [M+H] + .

[0258] Step 4: Synthesis of compound 11-5:

[0259] Substrate 11-4 (1.5 g, 4.74 mmol) was added to a dry single-necked bottle, Pd / C (172.73 mg, 1.42 mmol) was dissolved in MeOH / AcOH (10 mL), and H 2 Replace 3 times, H 2 The reaction was carried out at room temperature for 3 h and monitored by LC-MS. After the reaction was completed, the mixture was filtered through diatomaceous earth and the filtrate was concentrated under reduced pressure without purification. The crude product was directly used in the next step to obtain compound 11-5 (1.5 g, crude) as a black oil. LCMS (E+) m / z: 287.4 [M+H] + .

[0260] Step 5: Synthesis of compound 11:

[0261] Substrate 11-5 (35 mg, 122.20 μmol) was added to a dry single-mouth bottle, compound IM-5 (49.67 mg, 122.20 μmol) was dissolved in THF (3 mL), and PTSA (52.61 mg, 305.50 μmol) was added, the temperature was raised to 110°C, and the reaction was stirred for 5 hours, and LC-MS was monitored. After the reaction was completed, the organic solvent was concentrated under reduced pressure, extracted with water and ethyl acetate three times, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residual impurities were removed by MPLC to obtain compound 11 (21 mg, 32.56 μmol, 26.65% yield) as a green solid. 1 H NMR (600 MHz, DMSO-d 6 )δ10.13(s,1H),8.80(s,1H),7.80(s,1H),7.59(s,2H),7.30(d,J=7.7Hz,1H),6.91(d,J=8.5Hz,2H ),6.59(d,J=7.9Hz,1H),5.64(ddt,J=18.3,13.6,6.4Hz,1H),5.00(d,J=10.1Hz,1H),4.90(d,J=17 .0Hz,1H),4.75(s,2H),3.37(s,6H),3.13–3.09(m,6H),2.98–2.91(m,2H),2.80–2.63(m,4H),2.63 –2.55(m,2H),2.03–2.00(m,2H),1.77–1.60(m,2H),1.43–1.39(m,1H),1.29–1.25(m,1H).LCMS(ESI + )m / z:629.5[M+H] + ,HPLC method B:R T =5.94min,purity:97.5%

[0262] Example 12: Synthesis of Compound 12

[0263]

[0264] Step 1: Synthesis of compound 12-2:

[0265] Add substrate IM-2-1 (416 mg, 2.1 mmol) and DMSO (8 mL) to a dry single-mouth bottle, stir and dissolve, add potassium carbonate (387 mg, 2 mmol) and compound 12-1 (310 mg, 2 mmol), heat to 90 ° C for reaction, and monitor by LC-MS. After the reaction is completed, cool to room temperature, add water and stir until solid precipitates, filter out the solid, wash three times with water, wash three times with a small amount of methanol, and dry to obtain product 12-2 (200 mg, 30% yield), LCMS (ESI + )m / z:334.2[M+H] + .

[0266] Step 2: Synthesis of compound 12-3:

[0267] Substrate 12-2 (200 mg, 0.6 mmol) and DCM (5 mL) were added to a dry single-mouth bottle, and after stirring to dissolve, TFA (5 mL) was slowly added and monitored by LC-MS. After the reaction was completed, it was concentrated under reduced pressure. Methanol (5 mL) was added to the residue, and after stirring to dissolve, paraformaldehyde (180 mg) and acetic acid (500 μL) were added, and stirred at room temperature for 30 min, sodium cyanoborohydride (75 mg, 1.2 mmol) was added, and the temperature was raised to 50 ° C for reaction, and LC-MS was monitored. After the reaction was completed, it was cooled to room temperature, the solid was filtered off, the organic phase was concentrated under reduced pressure, and medium pressure liquid chromatography (alkali method) was purified to obtain compound 12-3 (120 mg, 81% yield), LCMS (ESI + )m / z:248.2[M+H] + .

[0268] Step 3: Synthesis of compound 12-4:

[0269] Substrate 12-3 (120 mg, 0.48 mmol) and methanol (5 mL) were added to a dry single-mouth bottle, stirred and dissolved, and Pd / C (12 mg) was added. The hydrogen was replaced five times, and the reaction was stirred at room temperature and monitored by LC-MS. After the reaction was completed, Pd / C was removed with diatomaceous earth, and the organic phase was concentrated by vacuum distillation to obtain product 12-4 (90 mg, 86.4% yield). LCMS (ESI + )m / z:218.2[M+H] + .

[0270] Step 4: Synthesis of compound 12:

[0271] Substrate IM-1 (52 mg, 0.15 mmol) and THF (3 mL) were added to a dry single-mouth bottle. After stirring and dissolving, m-CPBA (121 mg, 0.6 mmol, purity 85%) was added. The reaction was allowed to react at room temperature for 1 h and monitored by LC-MS. After the reaction was completed, DIPEA (98 mg, 758.28 μmol) was added. After stirring for 10 min, 12-4 (33 mg, 151.86 μmol) was added and monitored by LC-MS. After the reaction was completed, the mixture was concentrated under reduced pressure and the residue was purified by medium pressure liquid chromatography (alkaline method) to obtain compound 12 (13 mg, 15.38% yield). 1 H NMR (600 MHz, DMSO-d 6 )δ10.09(s,1H),8.82(s,1H),8.00-7.98(m,1H),7.78(d,J=8.1Hz,1H),7.61(d,J=7.7Hz,2H),7.28( dd,J=8.8,2.6Hz,1H),6.74(d,J=8.7Hz,1H),5.69-5.63(m,1H),5.32(s,1H),4.99(dd,J=10.3,1.5Hz ,1H),4.82(d,J=17.1Hz,1H),4.68(d,J=6.0Hz,2H),3.97(s,1H),3.39(s,1H),3.25–3.21(m,2H),2. 80-2.74(m,2H),2.32(s,3H),2.22(s,3H),1.83(d,J=9.3Hz,1H),1.72(d,J=9.2Hz,1H),1.46(s,6H). 1 H NMR (600 MHz, DMSO-d 6 ,D 2 O)δ8.82(s,1H),8.03-8.01(m,1H),7.76(d,J=8.1Hz,1H),7.65–7.57(m,2H),7.29(dd,J=8.7,2. 7Hz,1H),6.76(d,J=8.7Hz,1H),5.70-5.63(m,1H),5.03-5.01(m,1H),4.82(d,J=17.1Hz,1H),4.6 9(d,J=5.8Hz,2H),3.98(s,1H),3.42(s,1H),3.27–3.23(m,2H),2.81–2.74(m,2H),2.31(s,3H), 2.23(s,3H),1.86(d,J=9.5Hz,1H),1.75(d,J=9.4Hz,1H),1.47(s,6H).LCMS(E+)m / z:527.3[M+H] +,HPLC method B:R T =6.53min,purity:94.6%.

[0272] Example 13: Synthesis of Compound 13

[0273]

[0274] Step 1: Synthesis of compound 13-2:

[0275] Substrate 13-1 (1.03 g, 10 mmol) and DMSO (15 mL) were added to a dry single-mouth bottle, stirred and dissolved, potassium carbonate (2 g, 14 mmol) and p-fluoronitrobenzene (1.43 g, 10 mmol) were added, and the mixture was heated to 100 °C for reaction and monitored by LC-MS. After the reaction was completed, the mixture was cooled to room temperature, the solid was filtered out, and the filtrate was concentrated and purified by medium pressure liquid chromatography (acid method) to obtain compound 13-2 (2 g, 89% yield), LCMS (ESI + )m / z:225.1[M+H] + .

[0276] Step 2: Synthesis of compound 13-3:

[0277] Substrate 13-2 (44 mg, 0.2 mmol) and THF (2 mL) were added to a dry single-mouth bottle, and after stirring and dissolving, HATU (84 mg, 0.22 mmol) and DIPEA (28 mg, 0.22 mmol) were added. After stirring at room temperature for 30 min, methylamine hydrochloride (67 mg, 1 mmol) was added, and the temperature was raised to 65 ° C for reaction, and LC-MS monitoring was performed. After the reaction was completed, it was cooled to room temperature, extracted with ethyl acetate three times, and the organic phase was concentrated under reduced pressure to obtain a crude product 13-3 (24 mg, crude), LCMS (ESI + )m / z:238.3[M+H] + .

[0278] Step 3: Synthesis of compound 13-4:

[0279] Substrate 13-3 (24 mg, 0.1 mmol) and methanol (4 mL) were added to a dry single-mouth bottle, stirred and dissolved, and Pd / C (5 mg) was added. The hydrogen was replaced five times, and the reaction was stirred at room temperature and monitored by LC-MS. After the reaction was completed, Pd / C was removed by diatomaceous earth filtration, and the organic phase was concentrated by vacuum distillation to obtain a crude product 13-4 (18 mg, crude). LCMS (ESI + )m / z:208.2[M+H] + .

[0280] Step 4: Synthesis of compound 13:

[0281] Add substrate IM-1 (31 mg, 86 μmol) and THF (2 mL) to a dry single-mouth bottle, stir and dissolve, then add m-CPBA (26 mg, 129 μmol), react at room temperature for 1 h, and monitor by LC-MS. After the reaction is completed, add DIPEA (55 mg, 425 μmol), stir for 10 min, add 13-4 (18 mg, 85 μmol), heat to 80 °C for reaction, and monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure, and purify the residue by medium pressure liquid chromatography (alkaline method) to obtain compound 13 (7 mg, 16% yield). 1 H NMR (600 MHz, DMSO-d 6 )δ10.05(s,1H),8.79(s,1H),7.97-4.95(m,1H),7.75(d,J=4.4Hz,2H),7.60(d,J=7. 6Hz,1H),7.49(s,2H),6.59(d,J=8.0Hz,2H),5.69-5.62(m,1H),5.33(s,1H),5.04–4. 95(m,1H),4.98(d,J=10.0Hz,1H),4.82(d,J=17.0Hz,1H),4.67(d,J=5.9Hz,2H),3.78 (s,2H),3.41-3.40(m,2H),2.60(d,J=4.7Hz,3H),1.46(s,6H),1.08(t,J=7.0Hz,3H). 1 H NMR (600 MHz, DMSO-d 6 ,D 2 O)δ8.80(s,1H),8.00-7.97(m,1H),7.82(d,J=4.4Hz,1H),7.72(s,1H),7.61(d,J= 7.6Hz,1H),7.49(s,1H),6.59(d,J=8.2Hz,2H),5.69-5.63(m,1H),5.02(d,J=10.2 Hz,1H),4.83(d,J=17.0Hz,1H),4.68(s,2H),3.81(s,2H),3.42(q,J=7.0Hz,2H),2 .62(d,J=4.6Hz,3H),1.47(s,6H),1.08(t,J=7.0Hz,3H).LCMS(E+)m / z:517.3[M+H] + ,HPLC method B:R T =6.25min,purity:90.5%.

[0282] Example 14: Synthesis of Compound 14

[0283]

[0284] Step 1: Synthesis of compound 14-1:

[0285] Substrate 13-2 (44 mg, 0.2 mmol) and THF (2 mL) were added to a dry single-mouth bottle, and after stirring and dissolving, HATU (84 mg, 0.22 mmol) and DIPEA (28 mg, 0.22 mmol) were added. After stirring at room temperature for 30 min, N-methylpiperazine (40 mg, 0.4 mmol) was added, and the temperature was raised to 65 ° C for reaction, and LC-MS monitoring was performed. After the reaction was completed, it was cooled to room temperature, extracted with ethyl acetate three times, and the organic phase was concentrated under reduced pressure to obtain a crude product 14-1 (50 mg, crude), LCMS (ESI + )m / z:307.2[M+H] + Step 2: Synthesis of compound 14-2:

[0286] Substrate 14-1 (50 mg, 0.16 mmol) and methanol (4 mL) were added to a dry single-mouth bottle, stirred and dissolved, and Pd / C (5 mg) was added. The hydrogen gas was replaced five times, and the reaction was stirred at room temperature and monitored by LC-MS. After the reaction was completed, Pd / C was removed with diatomaceous earth, and the organic phase was concentrated under reduced pressure to obtain product 14-2 (40 mg, crude). LCMS (ESI + )m / z:277.2[M+H] + .

[0287] Step 3: Synthesis of compound 14:

[0288] Add substrate IM-1 (38 mg, 106 μmol) and THF (2 mL) to a dry single-mouth bottle, stir and dissolve, then add m-CPBA (40 mg, 200 μmol), react at room temperature for 1 h, and monitor by LC-MS. After the reaction is completed, add DIPEA (65 mg, 500 μmol), stir for 10 min, add compound 14-2 (40 mg, crude), heat to 80 °C for reaction, and monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure, and purify the residue by medium pressure liquid chromatography (alkaline method) to obtain compound 14 (12.2 mg, 17.58% yield). 1 H NMR (600 MHz, Methanol-d 4)δ8.76(s,1H),7.96-7.93(m,1H),7.76(s,1H),7.62(d,J=7.7Hz,1H),7.43(d,J=8.4Hz,2 H),6.68(d,J=8.6Hz,2H),5.73-5.67(m,1H),5.03(d,J=10.2Hz,1H),4.91(d,J=10.2Hz,1H ),4.80(d,J=5.8Hz,2H),4.59(s,2H),4.23(s,2H),3.76(s,2H),3.44(q,J=7.0Hz,2H),2.9 0-2.85(m,4H),2.62(s,3H),1.57(s,6H),1.18(t,J=7.0Hz,3H).LCMS(E+)m / z:586.4[M+H] + ,HPLC method B:R T =6.23min,purity:91.9%.

[0289] Example 15: Synthesis of Compound 15

[0290]

[0291] Step 1: Synthesis of compound 15-2:

[0292] Substrate 15-1 (2.44 g, 12.50 mmol), NBS (3.33 g, 18.70 mmol), AIBN (0.2 g, 1.21 mmol) were added to a single-mouth reaction bottle and heated with CCl 4 (20ml) was dissolved, and after nitrogen replacement 3 times, the reaction bottle was placed in an oil bath at 85°C and stirred overnight under nitrogen protection, and monitored by TLC. The solvent was removed by concentration under reduced pressure without purification, and the crude product was directly used in the next step to obtain the crude product 15-2 (3g, 11.03mmol, 88.1% yield).

[0293] Step 2: Synthesis of compound 15-3:

[0294] 4-Amino-1-methylpiperidine (1.8 g, 15.77 mmol) was added to the crude product of 15-2 (3 g, 11.03 mmol) to dissolve, and the reaction temperature was raised to 40 ° C. and stirred for 2 hours. LC-MS was monitored until the reaction was completed. The reactants were separated by column chromatography to obtain compound 15-3 (1.3 g, 4.72 mmol, 42.86% yield). LCMS (ESI + )m / z:276.1[M+H] + .

[0295] Step 3: Synthesis of compound 15-4:

[0296] Substrate 15-3 (0.6 g, 1.70 mmol) was added to the reaction flask and dissolved in anhydrous methanol (10 ml), and Pd / C (10% wt.) (60 mg) was added. After hydrogen replacement 3 times, the mixture was stirred at room temperature overnight and monitored by LC-MS. After the reaction was completed, the mixture was filtered using diatomaceous earth, and the filtrate was concentrated under reduced pressure and purified by column chromatography to obtain compound 15-4 (474 ​​mg, 1.92 mmol, 88.7% yield), LCMS (ESI + )m / z:246.2[M+H] + .

[0297] Step 4: Synthesis of compound 15:

[0298] Substrate IM-5 (39.3 mg, 0.097 mmol), substrate 15-4 (47.43 mg, 0.19 mmol), anhydrous acetonitrile (4 ml), TFA (55.18 mg, 0.48 mmol) were added to a microwave reaction tube. The reaction was carried out at 90 °C for 4 hours and monitored by LC-MS. After the reaction was completed, saturated NaHCO 3 The solution was quenched and extracted with ethyl acetate three times. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure and purified by medium pressure liquid chromatography to obtain the product 15 (10 mg, 0.017 mmol, 17.6% yield). 1 H NMR (600 MHz, Chloroform-d 6 )δ8.89(s,1H),8.03(s,1H),7.79(d,J=8.3Hz,1H),7.75(s,1H),7.71–7.68(m,1H ),7.56(d,J=8.2Hz,1H),7.31(d,J=7.7Hz,1H),6.74(d,J=8.0Hz,1H),5.69–5.63 (m,1H),5.05–4.96(m,2H),4.92(d,J=6.3Hz,2H),4.37(s,3H),3.34(s,6H),3.11 (s,2H),2.44(s,3H),2.33(s,2H),2.05(s,2H),1.89(d,J=12.5Hz,2H).LCMS(ESI + )m / z:588.1[M+H] + ,HPLC method B:R T =5.34min,purity:96.5%

[0299] Example 16: Synthesis of Compound 16

[0300]

[0301] Step 1: Synthesis of compound 16-2:

[0302] Pyridine (7.91 g, 100 mmol) and acetonitrile (40 mL) were added to a dry single-mouth bottle, stirred evenly, and 2-chloroacetamide (9.35 g, 100 mmol) was added, and the temperature was raised to 90°C for overnight reaction. After the reaction was completed, the solid was cooled to room temperature, filtered out, washed with petroleum ether three times, and recrystallized with ethanol to obtain the product 16-2 (10.8 g, 62% yield).

[0303] Step 2: Synthesis of compound 16-4:

[0304] Add substrate 16-2 (3.6 g, 21.15 mmol) and n-butanol (50 mL) to a dry single-mouth bottle, stir evenly, add 16-3 (1.82 g, 7.05 mmol), acetic acid (1.5 mL) and piperidine (2.5 mL), heat to 120 °C for reaction, and monitor with LC-MS. After the reaction is completed, filter out the solid, and the filtrate is treated with CHCl 3 (containing 5% MeOH) for three times, the organic phase was concentrated under reduced pressure, and the obtained solid was recrystallized from methanol to obtain the product 16-4 (1.92 g, 91% yield), LCMS (ESI + )m / z:298.2[M+H] + .

[0305] Step 3: Synthesis of compound 16-5:

[0306] Substrate 16-4 (500 mg, 1.68 mmol) and methanol (15 mL) were added to a dry single-mouth bottle, stirred evenly, Pd / C (50 mg) was added, hydrogen was replaced five times, the reaction was stirred at room temperature, and LC-MS was monitored. After the reaction was completed, Pd / C was removed with diatomaceous earth, washed with methanol, and the organic phase was concentrated under reduced pressure to obtain product 16-5 (310 mg, 69% yield), LCMS (ESI + )m / z:268.1[M+H] + .

[0307] Step 4: Synthesis of compound 16:

[0308] Add substrate IM-1 (70 mg, 200 μmol) and tetrahydrofuran (4 mL) to a dry single-mouth bottle, stir and dissolve, then add m-CPBA (60 mg, 300 μmol, purity 85%), react at room temperature for 1 h, and monitor by LC-MS. After the reaction is completed, add DIPEA (129.24 mg, 1.00 mmol), stir for 10 min, add 16-5 (53.47 mg, 200.00 μmol), microwave at 80 ° C, and monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure, and the residue is purified by medium pressure liquid chromatography (alkaline method) to obtain compound 16 (6.3 mg, 5.22% yield). 1 H NMR (600 MHz, DMSO-d 6 )δ11.81(s,1H),10.43(s,1H),8.93(s,1H),8.08-8.05(m,1H),7.81-7.78(m,3H ),7.64(d,J=7.7Hz,1H),7.58(d,J=8.2Hz,2H),6.22(s,1H),5.71-5.65(m,1H),5 .35(s,1H),5.01(d,J=10.2Hz,1H),4.84(d,J=17.2Hz,1H),4.69(d,J=5.9Hz,2H ),3.47(s,2H),3.09(s,1H),3.00(s,2H),1.84(s,2H),1.56(s,2H),1.47(s,6H). 1 H NMR (600 MHz, DMSO-d 6 ,D 2 O)δ8.93(s,1H),8.09-8.06(m,1H),7.81-7.76(m,3H),7.65(d,J=7.6Hz,1H) 7.59(d,J=8.4Hz,2H),6.27(s,1H),5.71-5.64(m,1H),5.04(d,J=10.2Hz,1H ),4.84(d,J=17.1Hz,1H),4.70(d,J=5.8Hz,2H),3.93(s,2H),3.16(s,1H),3 .02(s,2H),1.88(s,2H),1.57(s,2H),1.48(s,6H).LCMS(E+)m / z:577.3[M+H] + ,HPLCmethod B:R T =5.96min,purity:95.5%.

[0309] Example 17: Synthesis of Compound 17

[0310]

[0311] Step 1: Synthesis of compound 17-3

[0312] In a dry round-bottom flask, 17-2 (130 mg, 0.88 mmol) and DIPEA (0.58 g, 4.4 mmol) were dissolved in DMSO (1 ml) in sequence, and then 17-1 (1.1 mmol, 148 mmol) was added. Stir at room temperature for 2 hours. LC-MS monitoring, after the reaction was completed, diluted with water, extracted with ethyl acetate three times, washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and directly put into the next step without purification, to obtain a crude product 17-3 (150 mg, 0.64 mmol, 65.6% yield), LCMS (ESI+) m / z: 235.1 [M+H]+.

[0313] Step 2: Synthesis of compound 17-4

[0314] 17-3 (150 mg, 0.64 mmol), catalytic amount of Pd / C (45 mg) and MeOH (3 mL) were added to a 50 mL round-bottom flask, and the mixture was replaced with hydrogen three times and stirred vigorously at room temperature for 12 hours under a hydrogen atmosphere. LC-MS monitoring, diatomaceous earth filtration, the filtrate was concentrated under reduced pressure and purified by medium pressure liquid chromatography to obtain compound 17-4 (100 mg, 0.49 mmol, 76.1% yield), LCMS (ESI+) m / z: 205.1 [M+H]+.

[0315] Step 3: Synthesis of compound 17

[0316] m-CPBA (39 mg, 0.18 mmol) was added to a THF (1 mL) solution containing IM-1 (51 mg, 0.18 mmol) and stirred at room temperature for 10 minutes. After the disappearance of the starting material, DIPEA (122 mg, 0.75 mmol) was added and stirred at room temperature for 20 minutes. Subsequently, 17-4 (51 mg, 0.15 mmol) was added and stirred at room temperature for 1 hour. After the reaction was completed, water was added and extracted with ethyl acetate 3 times. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by acidic preparative high performance liquid chromatography to obtain compound 17 (26 mg, 0.05 mmol, 35.5% yield). 1 H NMR (600 MHz, DMSO-d 6)δ10.13(s,1H),8.81(s,1H),8.04(s,1H),7.74(d,J=7.6Hz,1H),7.60(d,J=7.7H z,1H),7.57(s,1H),6.82(d,J=8.5Hz,2H),5.69-5.64(m,1H),5.32(s,1H),4.99(d ,J=10.2Hz,1H),4.82(d,J=17.1Hz,1H),4.68(s,2H),4.42(s,2H),2.82–2.72(m,2 H),1.84(s,4H),1.49(s,1H),1.46(s,6H),1.38(s,2H).LCMS(E+)m / z:514.5[M+H] + ,HPLC method A:R T =7.13min,purity:>99.9%.

[0317] Example 18: Synthesis of Compound 18

[0318]

[0319] Step 1: Synthesis of compound 18-2

[0320] Add 18-1 (4 g, 20 mmol) to a dry single-mouth bottle and dissolve it in DCM (50 ml). Add TFA (25 mL) at room temperature and stir for 30 minutes. Monitor by LC-MS. After the reaction is completed, remove the solvent in vacuo, dilute with water, extract with ethyl acetate three times, dry the organic phase with anhydrous sodium sulfate and concentrate under reduced pressure. The crude product 18-2 is directly used in the next step without purification (1.8 g, crude). LCMS (ESI+) m / z: 100.1 [M+H] +

[0321] Step 2: Synthesis of compound 18-4

[0322] In a dry single-necked bottle, 18-2 (1 g, 7 mmol) was dissolved in DMSO (10 mL), and 18-3 (1 g, 10.5 mmol), K 2 CO 3 (4.8 g, 35 mmol). The reaction mixture was stirred at room temperature overnight. LC-MS monitoring, after the reaction was completed, it was diluted with 100 mL of water, extracted with ethyl acetate three times, the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and the obtained crude product 18-4 was directly used in the next step without purification (1.35 g, crude). LCMS (ESI+) m / z: 221.1 [M+H] +

[0323] Step 3: Synthesis of compound 18-6

[0324] In a dry round-bottom flask, 18-4 (1.35 g, 6.2 mmol) and 18-5 (1.47 g, 7.4 mmol) were dissolved in methanol (15 ml), and acetic acid (1 ml) was added and reacted at room temperature for 30 minutes. Then sodium cyanoborohydride (584 mg, 9.3 mmol) was added, the temperature was raised to 60°C, and stirred for 4 hours. LC-MS monitoring, after the reaction was completed, the solvent was removed under reduced pressure, water was added, and the mixture was extracted with ethyl acetate three times. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product 18-6 (1.75 g, crude), LCMS (ESI+) m / z: 403.2 [M+H] +

[0325] Step 4: Synthesis of compound 18-7

[0326] 18-6 (1.75 g, 4.4 mmol), catalytic amount of Pd / C (200 mg) and MeOH (20 mL) were added to a 50 mL round-bottom flask. Stir at room temperature for 12 hours under a hydrogen atmosphere. Monitor by LC-MS. After the reaction, the reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain compound 18-7 (1.46 g, 3.9 mmol, 90.2% yield). LCMS (ESI+) m / z: 373.3 [M+H] + .

[0327] Step 5: Synthesis of compound 18-8

[0328] In a dry round-bottom flask, 18-7 (1.46 g, 3.9 mmol) was dissolved in THF and heated at 0 °C under N 2 Slowly add 2M LiAlH 4 The reaction temperature was raised to 65°C and stirred for 6 hours at this temperature, monitored by LC-MS. After the reaction was completed, the reaction temperature was lowered to 0°C, and water (0.76 ml), 10% NaOH aqueous solution (0.76 ml), and water quenching (2.3 ml) were added in sequence. Stirred at room temperature for 30 minutes, anhydrous Na 2 SO 4 Dry and filter with celite. The filtrate was concentrated in vacuo and purified by column chromatography to obtain compound 18-8 (736 mg, 2.5 mmol, 65.6% yield). LCMS (ESI+) m / z: 287.2 [M+H] + .

[0329] Step 6: Synthesis of compound 18

[0330] According to the synthesis method of step 3 in Example 17, 17-4 (51 mg, 0.15 mmol) in step 3 was replaced by 18-8 (42.1 mg, 0.15 mmol) and the synthesis method was the same to obtain compound 18 (20 mg, 33.6 μmol, 22.9% yield). 1 H NMR (600 MHz, DMSO-d 6 )δ10.19(s,1H),8.84(s,1H),8.04(s,1H),7.76(d,J=8.1Hz,1H),7.62(d,J=7.7Hz,3H),7.00 (d,J=8.5Hz,3H),5.70–5.63(m,1H),5.00(d,J=10.2Hz,1H),4.83(d,J=17.2Hz,1H),4.68(s,3 H), 4.45 (s, 1H), 3.82 (s, 6H), 3.37 (s, 2H), 2.92 (s, 3H), 2.69 (t, J = 12.4Hz, 2H), 2.23 (d, J = 64. 4Hz,2H),2.07(d,J=11.8Hz,1H),1.78–1.63(m,2H),1.47(s,6H).LCMS(ESI+)m / z:596.4[M+H] + HPLC method A:R T =6.35min,purity:96.9%

[0331] Example 19: Synthesis of Compound 19

[0332]

[0333] Step 1: Synthesis of compound 19-2:

[0334] Substrate 19-1 (200 mg, 0.1 mmol) and DMSO (5 mL) were added to a dry single-mouth bottle. Potassium carbonate (470 mg, 3.4 mmol) and p-fluoronitrobenzene (141 mg, 1 mmol) were added under stirring, and the temperature was raised to 110°C for reaction, and LC-MS was monitored. After the reaction was completed, the mixture was cooled to room temperature, iodomethane (141 mg, 1 mmol) was added, and the temperature was raised to 90°C for reaction, and LC-MS was monitored. After the reaction was completed, the mixture was cooled to room temperature, the solid was filtered out, and the filtrate was concentrated and purified by medium pressure liquid chromatography (alkaline method) to obtain product 19-2 (50 mg, 19% yield), LCMS (ESI + )m / z:262.3[M+H] + .

[0335] Step 2: Synthesis of compound 19-3:

[0336] Substrate 19-2 (50 mg, 0.19 mmol) and methanol (2 mL) were added to a dry single-mouth bottle, stirred and dissolved, and Pd / C (10 mg) was added. The hydrogen was replaced five times, and the reaction was stirred at room temperature and monitored by LC-MS. After the reaction was completed, Pd / C was removed with diatomaceous earth, and the organic phase was concentrated by vacuum distillation to obtain product 19-3 (40 mg, crude). LCMS (ESI + )m / z:232.2[M+H] + .

[0337] Step 3: Synthesis of compound 19:

[0338] Add substrate IM-1 (52 mg, 0.15 mmol) and tetrahydrofuran (3 mL) to a dry single-mouth bottle, stir and dissolve, then add m-CPBA (121 mg, 0.6 mmol, purity 85%), react at room temperature for 1 h, and monitor by LC-MS. After the reaction is completed, add DIPEA (95.17 mg, 736.42 μmol), stir for 10 min, then add 19-3 (40 mg, crude), and monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure, and purify the residue by medium pressure liquid chromatography (alkaline method) to obtain compound 19 (35 mg, 40.96% yield). 1 H NMR (600 MHz, DMSO-d 6 )δ10.11(s,1H),8.80(s,1H),7.99-7.96(m,1H),7.75(s,1H),7.62(d,J=7.7Hz,1H),7. 47(s,2H),6.62(d,J=8.3Hz,2H),6.19(d,J=23.4Hz,1H),5.69-5.63(m,1H),4.99(d,J= 10.2Hz,1H),4.83(d,J=17.1Hz,1H),4.67(s,2H),3.89(s,2H),3.51(s,2H),3.29(s,3H ),2.97(s,3H),2.22(s,2H),2.00(d,J=7.8Hz,2H),1.76(d,J=14.1Hz,1H),1.46(s,6H). 1 H NMR (600 MHz, DMSO-d 6 ,D 2O)δ8.81(s,1H),8.01-7.98(m,1H),7.74(s,1H),7.62(d,J=7.8Hz,1H),7.49(s,2H), 6.64(d,J=8.4Hz,2H),5.69-5.63(m,1H),5.02(d,J=10.2Hz,1H),4.83(d,J=17.0Hz,1 H),4.68(s,2H),3.90-3.88(m,2H),3.43-3.42(m,4H),3.14–3.12(m,1H),2.97(s,3H) ,2.23–2.20(m,2H),2.03(s,2H),1.82(s,1H),1.47(s,6H).LCMS(E+)m / z:541.3[M+H] + ,HPLC method B:R T =5.65min,purity:93.2%.

[0339] Example 20: Synthesis of Compound 20

[0340]

[0341] Step 1: Synthesis of compound 20-2:

[0342] Substrate 20-1 (136 mg, 1 mmol) and DMSO (8 mL) were added to a dry single-mouth bottle, stirred and dissolved, potassium carbonate (332 mg, 2.4 mmol) and p-fluoronitrobenzene (141 mg, 1 mmol) were added, and the mixture was heated to 90°C for reaction and monitored by LC-MS. After the reaction was completed, the mixture was cooled to room temperature, and water was added and stirred to precipitate solids. The solids were filtered out, washed three times with water, washed three times with a small amount of methanol, and dried to obtain product 20-2 (180 mg, 81.8% yield), LCMS (ESI + )m / z:221.2[M+H] + .

[0343] Step 2: Synthesis of compound 20-3:

[0344] Substrate 20-2 (180 mg, 0.82 mmol) and methanol (7 mL) were added to a dry single-mouth bottle, and Pd / C (18 mg) was added under stirring. The hydrogen gas was replaced five times, and the reaction was stirred at room temperature and monitored by LC-MS. After the reaction was completed, Pd / C was removed with diatomaceous earth, and the organic phase was concentrated under reduced pressure to obtain a crude product 20-3 (120 mg, crude), LCMS (ESI+) m / z: 191.1 [M+H]+.

[0345] Step 3: Synthesis of compound 20:

[0346] Add substrate IM-1 (52 mg, 0.15 mmol) and tetrahydrofuran (3 mL) to a dry single-mouth bottle, stir and dissolve, then add m-CPBA (121 mg, 0.6 mmol, purity 85%), react at room temperature for 1 h, and monitor by LC-MS. After the reaction is completed, add DIPEA (98 mg, 758.28 μmol), stir for 10 min, and then add compound 20-3 (46 mg, 151.86 μmol), and monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure, and the residue is purified by medium pressure liquid chromatography (alkaline method) to obtain compound 20 (25 mg, 0.05 mmol, 33.84% yield). 1 H NMR (600 MHz, DMSO-d 6 )δ10.07(s,1H),8.80(s,1H),8.02-8.01(m,1H),7.75(s,1H),7.60(d,J=7.6Hz,1H),7.53–7.38(m ,2H),6.62(d,J=7.2Hz,2H),5.70-5.63(m,1H),5.33(s,1H),5.00(d,J=10.2Hz,1H),4.83(d,J=17. 1Hz,1H),4.69(s,2H),4.60(s,1H),4.53(s,1H),3.75(d,J=6.8Hz,1H),3.67(d,J=7.4Hz,1H),3.50 (d,J=9.0Hz,1H),2.95(d,J=9.0Hz,1H),1.93(d,J=9.2Hz,1H),1.84(d,J=8.8Hz,1H),1.47(s,6H). 1 HNMR (600 MHz, DMSO-d 6 ,D 2 O)δ8.82(s,1H),8.05-8.02(m,1H),7.74(s,1H),7.62(d,J=7.6Hz,1H),7.52(s,2H),6.63(d,J=7 .6Hz,2H),5.70-5.64(m,1H),5.03(d,J=10.2Hz,1H),4.84(d,J=17.0Hz,1H),4.70(s,2H),4.62(s ,1H),4.51(s,1H),3.78(d,J=7.4Hz,1H),3.68(d,J=7.6Hz,1H),3.52(d,J=9.4Hz,1H),2.96(d,J =9.2Hz,1H),1.95(d,J=9.0Hz,1H),1.86(d,J=9.0Hz,1H),1.48(s,6H).LCMS(E+)m / z:500.1[M+H]+ ,HPLC method B:R T =6.69min,purity:99.6%.

[0347] Example 21: Synthesis of Compound 21

[0348]

[0349] Step 1: Synthesis of compound 21-3:

[0350] Substrate 21-1 (3.0 g, 19.34 mmol) and DMSO (20 mL) were added to a dry single-mouth bottle, stirred and dissolved, potassium carbonate (2.67 g, 19.34 mmol) and 21-2 (3.82 g, 19.34 mmol) were added, and the mixture was heated to 100 °C for 3 hours and monitored by LC-MS. After the reaction was completed, the mixture was cooled to room temperature, and water was added and stirred to precipitate solids. The solids were filtered out, washed three times with water, washed three times with a small amount of methanol, and dried to obtain product 21-3 (6 g, 18.00 mmol, 93.06% yield), LCMS (ESI + )m / z:334.4[M+H] + .

[0351] Step 2: Synthesis of compound 21-4:

[0352] Substrate 21-3 (6.67 g, 20.01 mmol), TFA (2.28 g, 20.01 mmol) and DCM (50 mL) were added to a dry single-mouth bottle and stirred to dissolve. The mixture was reacted at room temperature for 18 hours and monitored by LC-MS. After the reaction was completed, the mixture was concentrated under reduced pressure, and water and ethyl acetate were added to extract three times. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product 21-4 (4 g, crude). LCMS (ESI + )m / z:234.3[M+H] + .

[0353] Step 3: Synthesis of compound 21-6:

[0354] In a dry single-mouth bottle, add substrate 21-4 (3 g, 12.86 mmol), compound 21-5 (1.46 g, 12.86 mmol), NaBH 3CN (1.2 g, 19.11 mmol) and methanol (30 mL) were stirred and dissolved, and the temperature was raised to 60°C for 18 hours, and LC-MS was used for monitoring. After the reaction was completed, the mixture was concentrated under reduced pressure, and water and ethyl acetate were added for extraction three times. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product 21-6 (3 g, 9.08 mmol, 70.60% yield) was obtained by medium pressure liquid chromatography. LCMS (ESI + )m / z:331.4[M+H] + .

[0355] Step 4: Synthesis of compound 21-7:

[0356] Substrate 21-6 (200 mg, 0.61 mmol) and methanol (10 mL) were added to a dry single-mouth bottle, stirred evenly, Pd / C (40 mg) was added, hydrogen was replaced five times, the reaction was stirred at room temperature, and LC-MS was monitored. After the reaction was completed, Pd / C was removed with diatomaceous earth, washed with methanol, and the organic phase was concentrated under reduced pressure to obtain product 21-7 (50 mg, 27.5% yield), LCMS (ESI + )m / z:301[M+H] + .

[0357] Step 5: Synthesis of compound 21:

[0358] Add substrate IM-1 (17.8 mg, 50.08 μmol) and tetrahydrofuran (2 mL) to a dry single-mouth bottle, stir and dissolve, then add m-CPBA (12.9 mg, 74.74 μmol), react at room temperature for 1 h, and monitor by LC-MS. After the reaction is completed, add DIPEA (6.47 mg, 50.08 μmol, 8.72 μL), stir for 10 min, and then add compound 21-7 (15.05 mg, 50.08 μmol), and monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure, and the residue is purified by medium pressure liquid chromatography (alkali method) to obtain compound 21 (5 mg, 8.16 μmol, 16.29% yield). 1 H NMR (600 MHz, DMSO-d 6)δ10.09(s,1H),8.82(s,1H),7.99–7.97(m,1H),7.78(d,J=7.8Hz,1H),7.60(d,J=7.8Hz,1H),7.57(s,1H),7. 29–7.27(m,1H),6.72(d,J=9.0Hz,1H),5.69–5.63(m,1H),5.33(s,1H),4.99(d,J=10.2Hz,1H),4.82(d,J=17. 4Hz,1H),4.68(d,J=5.9Hz,2H),3.95(s,1H),3.66(s,1H),3.20(q,J=8.4Hz,2H),3.02(d,J=8.4Hz,1H),2.68( d,J=7.8Hz,2H),2.56(d,J=9.0Hz,1H),2.22(s,3H),2.16(s,1H),2.12(s,3H),1.91(s,1H),1.88(s,1H),1.74 -1.69(m,4H),1.46(s,6H),1.38–1.25(m,2H).LCMS(ESI + )m / z:610.6[M+H] + .HPLC method A:R T =4.45min,purity:99.5%.

[0359] Example 22: Synthesis of Compound 22

[0360]

[0361] Step 1: Synthesis of compound 22-2:

[0362] Substrate 22-1 (158 mg, 499.31 μmol) was added to a dry single-mouth bottle, TFA (3 mL) and DCM (3 mL) were stirred and dissolved, and the mixture was reacted at room temperature for 18 hours and monitored by LC-MS. After the reaction was completed, the mixture was concentrated under reduced pressure, and water and ethyl acetate were added to extract three times. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product 22-2 (100 mg, crude), LCMS (ESI + )m / z:217.3[M+H] + .

[0363] Step 2: Synthesis of compound 22-3:

[0364] Substrate 22-2 (100 mg, 462.28 μmol) and DMSO (4 mL) were added to a dry single-mouth bottle, stirred and dissolved, potassium carbonate (97 mg, 701.88 μmol) and compound 4-2 (72 mg, 510.28 μmol) were added, and heated to 100 ° C for 3 hours, and monitored by LC-MS. After the reaction was completed, it was cooled to room temperature, and water was added and stirred to precipitate solids. The solids were filtered out, washed three times with water, washed three times with a small amount of methanol, and dried to obtain product 22-3 (150 mg, 444.56 μmol, 96.17% yield), LCMS (ESI + )m / z:338.4[M+H] + .

[0365] Step 3: Synthesis of compound 22-4:

[0366] Substrate 22-3 (50 mg, 148.19 μmol) and methanol (4 mL) were added to a dry single-mouth bottle, stirred evenly, Pd / C (8 mg) was added, hydrogen was replaced five times, the reaction was stirred at room temperature, and LC-MS was monitored. After the reaction was completed, Pd / C was removed with diatomaceous earth, washed with methanol, and the organic phase was concentrated under reduced pressure to obtain product 22-4 (20 mg, 65.06 μmol, 43.90% yield), LCMS (ESI + )m / z:308.4[M+H] + .

[0367] Step 4: Synthesis of compound 22:

[0368] Substrate 22-4 (24.70 mg, 80.34 μmol) and tetrahydrofuran (2 mL) were added to a dry single-mouth bottle, stirred to dissolve, and DIPEA (51.91 mg, 401.68 μmol, 69.96 μL) and compound IM-6 (30 mg, 80.34 μmol) were added, and the reaction was carried out at room temperature for 1 h, and monitored by LC-MS. After the reaction was completed, the mixture was concentrated under reduced pressure, and the residue was purified by medium pressure liquid chromatography to obtain compound 22 (5 mg, 7.30 μmol, 9.09% yield). 1 H NMR (600 MHz, DMSO-d 6)δ10.10(s,1H),8.80(s,1H),8.02(s,1H),7.79-7.78(m,1H),7.61-7.55(m,3H),7.18-7.17(m,5H) ,6.77(d,J=8.4Hz,2H),5.70–5.63(m,1H),5.33(s,1H),5.00(d,J=10.3Hz,1H),4.83(d,J=17.2Hz, 1H),4.68(s,2H),3.59(d,J=11.3Hz,2H),3.38(s,2H)3.08(d,J=11.3Hz,2H),2.87(d,J=10.6Hz,2H ),2.26(d,J=10.7Hz,2H),2.09(s,2H),1.70(d,J=12.2Hz,1H),1.60(d,J=12.1Hz,1H),1.47(s,6H). 1 H NMR (600 MHz, DMSO-d 6 ,D 2 O)δ8.81(s,1H),8.03(s,1H),7.78(s,1H),7.61-7.58(m,3H),7.18(s,5H),6.78(d,J=8.6H z,2H),5.69-5.63(m,1H),5.02(d,J=10.3Hz,1H),4.83(d,J=17.1Hz,1H),4.68(s,2H),3.64 (s,2H),3.39(s,2H),3.08(d,J=10.9Hz,2H),2.87(d,J=10.8Hz,2H),2.28(s,2H),2.10(s, 2H),1.70(d,J=11.9Hz,1H),1.60(d,J=12.2Hz,1H),1.47(s,6H).LCMS(E+)m / z:617.4[M+H] + ,HPLC method B:R T =6.01min,purity:90.1%.

[0369] Example 23: Synthesis of Compound 23

[0370]

[0371] Step 1: Synthesis of compound 23-1:

[0372] Substrate 22-1 (200 mg, 632.04 μmol) and methanol (5 mL) were added to a dry single-mouth bottle, stirred evenly, Pd / C (20 mg) was added, hydrogen was replaced five times, the reaction was stirred at room temperature, and LC-MS was monitored. After the reaction was completed, Pd / C was removed with diatomaceous earth, washed with methanol, and the organic phase was concentrated under reduced pressure to obtain product 23-1 (140 mg, crude), LCMS (ESI + )m / z:227.3[M+H] + .

[0373] Step 2: Synthesis of compound 23-2:

[0374] Substrate 23-1 (128.31 mg, 566.97 μmol) and DMSO (3 mL) were added to a dry single-mouth bottle, stirred and dissolved, potassium carbonate (96.74 mg, 0.7 mmol) and p-fluoronitrobenzene (4-2, 80 mg, 566.97 μmol) were added, and heated to 90 ° C for 6 hours, and monitored by LC-MS. After the reaction was completed, it was cooled to room temperature, and water was added and stirred to precipitate solids. The solids were filtered out, washed three times with water, washed three times with a small amount of methanol, and dried to obtain the product 23-2 (120 mg, crude), LCMS (ESI + )m / z:348.4[M+H] + .

[0375] Step 3: Synthesis of compound 23-3:

[0376] Substrate 23-2 (69 mg, 198.61 μmol) was added to a dry single-mouth bottle, and TFA (3 mL) and DCM (3 mL) were stirred and dissolved. The mixture was reacted at room temperature for 18 hours and monitored by LC-MS. After the reaction was completed, the mixture was concentrated under reduced pressure, and water and ethyl acetate were added to extract three times. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product 23-3 (49 mg, crude). LCMS (ESI + )m / z:248.3[M+H] + .

[0377] Step 4: Synthesis of compound 23-4:

[0378] Substrate 23-3 (49.46 mg, 0.2 mmol) was added to a dry single-mouth bottle, methanol (4 mL) was added and stirred to dissolve, formaldehyde (34.06 mg, 1.00 mmol) and acetic acid (11 mL) were added, and after stirring for 10 minutes, NaCNBH 3(25.14 mg, 400.00 μmol), heated to 50 ° C for 2 hours, monitored by LC-MS. After the reaction, it was concentrated under reduced pressure, extracted three times with water and ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was purified by medium pressure liquid chromatography to obtain compound 23-4 (30 mg, 114.80 μmol, 57.40% yield), LCMS (ESI + )m / z:262.3[M+H] + .

[0379] Step 5: Synthesis of compound 23-5:

[0380] Substrate 23-4 (30 mg, 114.80 μmol) and methanol (3 mL) were added to a dry single-mouth bottle, stirred evenly, Pd / C (8 mg) was added, hydrogen was replaced five times, the reaction was stirred at room temperature, and LC-MS was monitored. After the reaction was completed, Pd / C was removed with diatomaceous earth, washed with methanol, and the organic phase was concentrated under reduced pressure to obtain product 23-5 (20 mg, crude), LCMS (ESI + )m / z:232.3[M+H] + .

[0381] Step 6: Synthesis of compound 23:

[0382] Substrate 23-5 (18.58 mg, 80.34 μmol) and tetrahydrofuran (2 mL) were added to a dry single-mouth bottle, stirred to dissolve, and DIPEA (51.91 mg, 401.68 μmol, 69.96 μL) and compound IM-6 (30 mg, 80.34 μmol) were added. The reaction was carried out at room temperature for 1 h and monitored by LC-MS. After the reaction was completed, the mixture was concentrated under reduced pressure and the residue was purified by medium pressure liquid chromatography to obtain compound 23 (8 mg, 13.58 μmol, 16.91% yield). 1 H NMR (600 MHz, DMSO-d 6)δ10.09(s,1H),8.80(s,1H),8.04(s,1H),7.76(d,J=8.0Hz,1H),7.61-7.56(m,3H),6.7 3(d,J=8.6Hz,2H),5.65-5.63(m,1H),5.33(s,1H),4.99(dd,J=10.3,1.6Hz,1H),4.83(d d,J=17.1,1.7Hz,1H),4.68(d,J=6.1Hz,2H),3.57(d,J=11.2Hz,2H),3.46(s,3H),3.00( dd,J=11.3,4.8Hz,2H),2.88(s,2H),2.17-2.09(m,4H),1.66-1.54(m,2H),1.46(s,6H). 1 H NMR (600 MHz, DMSO-d 6 ,D 2 O)δ8.80(s,1H),8.05(s,1H),7.76(s,1H),7.62–7.55(m,3H),6.75(s,2 H),5.67-5.63(m,1H),5.02-5.01(m,1H),4.84–4.82(m,1H),4.68(d,J=6 .1Hz,2H),3.71(s,3H),3.58(d,J=11.4Hz,2H),2.99-2.89(m,4H),2.12 -2.04(m,4H),1.66–1.57(m,2H),1.47(s,6H).LCMS(E+)m / z:541.3[M+H] + ,HPLC method B:R T =5.29min,purity:94.1%.

[0383] Example 24: Synthesis of Compound 24

[0384]

[0385] Step 1: Synthesis of compound 24-2:

[0386] Add substrate 24-1 (2 g, 10.25 mmol) and carbon tetrachloride (40 mL) to a dry single-mouth bottle, stir evenly, add NBS (2.74 g, 15.37 mmol) and AIBN (269.24 mg, 1.64 mmol), replace with nitrogen five times, heat to 70 ° C, stir and reflux for 12 hours, and monitor with LC-MS. After the reaction is completed, concentrate under reduced pressure, and purify the residue by medium pressure liquid chromatography to obtain 24-2 (1.3 g, 4.74 mmol, 46.3% yield), LCMS (ESI+ )m / z:274.1[M+H] + .

[0387] Step 2: Synthesis of compound 24-4:

[0388] Substrate 24-2 (200 mg, 0.73 mmol) and 24-3 (166.42 mg, 1.46 mmol) were added to a dry single-mouth bottle, heated to 50°C, stirred and refluxed for 1 hour, and monitored by TLC. After the reaction was completed, it was concentrated under reduced pressure, a small amount of ethyl acetate was added, the solid was filtered out, and the filtrate was concentrated under reduced pressure. The residue was purified by medium pressure liquid chromatography to obtain 24-4 (180 mg, 0.65 mmol, 89.67% yield), LCMS (ESI + )m / z:276.1[M+H] + .

[0389] Step 3: Synthesis of compound 24-5:

[0390] Substrate 24-4 (180 mg, 0.65 mmol) and methanol (3 mL) were added to a dry single-mouth bottle, stirred evenly, Pd / C (36 mg) was added, hydrogen was replaced five times, the reaction was stirred at room temperature, and LC-MS was monitored. After the reaction was completed, Pd / C was removed with diatomaceous earth, washed with methanol, and the organic phase was concentrated under reduced pressure to obtain product 24-5 (100 mg, crude), LCMS (ESI + )m / z:246.3[M+H] + Step 4: Synthesis of compound 24-6:

[0391] Substrate IM-3 (0.2 g, 0.51 mmol) was added to a single-mouth reaction bottle, anhydrous THF (5 ml) was stirred and dissolved, and m-CPBA (208.2 g, 1.02 mmol, purity 85%) was added, and stirred at room temperature for 40 minutes, and monitored by LC-MS. The reactants were concentrated under reduced pressure and separated by medium pressure liquid chromatography to obtain compound 24-6 (130 mg, 0.31 mmol, 60.0% yield), LCMS (ESI + )m / z:423.5[M+H] + .

[0392] Step 5: Synthesis of compound 24:

[0393] Substrate 24-6 (31 mg, 73.38 μmol), 24-5 (36.00 mg, 146.75 μmol), TsOH (50.48 mg, 293.50 μmol) and anhydrous ACN (0.3 mL) were added to a dry microwave tube, stirred and dissolved, sealed, microwave heated to 110 ° C for 4 hours, and monitored by LC-MS. After the reaction was completed, it was cooled to room temperature, quenched by adding saturated sodium bicarbonate solution, extracted with ethyl acetate three times, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by medium pressure liquid chromatography to obtain product 24 (1.5 mg, 2.55 μmol, 3.48% yield). 1 H NMR (600 MHz, DMSO-d 6 )δ10.52(s,1H),8.91(s,1H),8.46(s,1H),7.95(s,1H),7.77(dd,J=8.2,2.2Hz,1H),7.55(d,J=8.3Hz,1H), 7.40(d,J=7.9Hz,1H),6.64(d,J=8.0Hz,1H),5.66(ddt,J=16.4,10.2,6.0Hz,1H),5.02(dd,J=10.3,1.6Hz,1 H),4.90(dd,J=17.1,1.6Hz,1H),4.80(s,2H),4.42(s,2H),4.08–3.97(m,1H),3.38(s,6H),2.89(d,J=11.1 Hz,2H),2.22(s,3H),2.07(t,J=11.8Hz,2H),1.82(dd,J=12.1,3.8Hz,2H),1.71(d,J=13.3Hz,2H).LCMS(ESI + )m / z:588.1[M+H] + ,HPLC method B:R T =5.22min,purity:92.3%

[0394] Example 25: Synthesis of Compound 25

[0395]

[0396] Step 1: Synthesis of compound 25-2:

[0397] Substrate 25-1 (0.5 g, 2.36 mmol) and DMSO (10 mL) were added to a dry single-mouth bottle, stirred and dissolved, potassium carbonate (651.05 mg, 4.71 mmol) and p-fluoronitrobenzene (4-2, 332.33 mg, 2.36 mmol) were added, and the mixture was heated to 100 °C for 6 hours and monitored by LC-MS. After the reaction was completed, the mixture was cooled to room temperature, and water was added and stirred to precipitate solids. The solids were filtered out, washed three times with water, washed three times with a small amount of methanol, and dried to obtain product 25-2 (0.7 g, 2.10 mmol, 89.15% yield), LCMS (ESI + )m / z:333.4[M+H] + .

[0398] Step 2: Synthesis of compound 25-3:

[0399] Substrate 25-2 (0.5 g, 1.50 mmol) and methanol (10 mL) were added to a dry single-mouth bottle, stirred evenly, Pd / C (910 mg) was added, hydrogen was replaced five times, the reaction was stirred at room temperature, and LC-MS was monitored. After the reaction was completed, Pd / C was removed with diatomaceous earth, washed with methanol, and the organic phase was concentrated under reduced pressure to obtain product 25-3 (0.4 g, 1.32 mmol, 87.91% yield), LCMS (ESI + )m / z:304.2[M+H] + .

[0400] Step 3: Synthesis of compound 25-4:

[0401] Substrate 25-3 (150 mg, 0.5 mmol) and THF (4 mL) were added to a dry single-necked bottle, stirred and dissolved, and LiAlH 4 (112.57 mg, 2.97 mmol), heated to 65 ° C for 2 hours, LC-MS monitoring. After the reaction, cooled to room temperature, added water and stirred to precipitate solids, filtered out the solids, washed three times with water, washed three times with a small amount of methanol, and dried to obtain the product 25-4 (80 mg, 0.36 mmol, 74.46% yield), LCMS (ESI + )m / z:218.3[M+H] + .

[0402] Step 4: Synthesis of compound 25:

[0403] Add substrate IM-1 (21.70 mg, 99.88 μmol) and tetrahydrofuran (2 mL) to a dry single-mouth bottle, stir and dissolve, then add m-CPBA (25.86 mg, 149.82 μmol), react at room temperature for 1 h, and monitor by LC-MS. After the reaction is completed, add DIPEA (64.54 mg, 499.40 μmol, 86.98 μL), stir for 10 min, add 25-4 (21.70 mg, 99.88 μmol), and monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure, and the residue is purified by medium pressure liquid chromatography (alkaline method) to obtain compound 25 (9 mg, 16.06 μmol, 16.08% yield). 1 H NMR (600 MHz, DMSO-d 6 )δ10.07(s,1H),8.80(s,1H),8.03-8.01(m,1H),7.75(s,1H),7.60(d,J=7.8Hz,1H),7 .51(s,2H),6.80(d,J=8.4Hz,2H),5.70-5.99(m,1H),5.31(s,1H),5.00-4.98(m,1H),4 .84-4.81(m,1H),4.68(d,J=6.0Hz,2H),4.20(s,2H),2.47-2.44(m,2H),2.30-2.28(m ,2H),2.06(s,3H),1.88(d,J=6.6Hz,2H),1.83(d,J=6.0Hz,2H),1.46(s,6H).LCMS(ESI + )m / z:527.3[M+H] + .HPLC method B:R T :7.83min,purity:94.0%.

[0404] Example 26: Synthesis of Compound 26

[0405]

[0406] Step 1, synthesis of compound 26-3:

[0407] Substrate 26-1 (1.27 g, 12.72 mmol), 26-2 (2 g, 8.48 mmol), t-BuONa (1 g, 10.17 mmol) and toluene (15 mL) were added to a dry microwave tube and stirred to dissolve. 2 Deoxygenate for 10 min, add Pd 2 (dba) 3(778 mg, 0.85 mmol), BINAP (264.6 mg, 0.43 mmol), and N 2 Deoxygenate for 10 min, seal the tube, heat in an oil bath to 110°C for 12 hours, and monitor with LC-MS. After the reaction, cool to room temperature, filter with diatomaceous earth, and separate the filtrate with medium pressure liquid chromatography to obtain product 26-3 (1.57 g, 6.15 mmol, 72.58% yield). LCMS (ESI + )m / z:255.2[M+H] + .

[0408] Step 2, synthesis of compound 26-5:

[0409] Substrate 26-3 (650 mg, 2.55 mmol) was added to a dry single-necked bottle and dissolved with acetone (6 mL) and water (7 mL), and 26-4 (637.87 mg, 3.82 mmol) and Pd(OAc) were added. 2 (57.25 mg, 0.25 mmol) and sodium carbonate (542 mg, 2.55 mmol), replaced with nitrogen 3 times, heated to 50 ° C under nitrogen protection for 12 hours, and monitored by LC-MS. After the reaction, it was cooled to room temperature, and water and ethyl acetate were added for extraction three times. The organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The residual impurities were purified by medium pressure liquid chromatography to obtain the product 26-5 (240 mg, 807.13 μmol, 31.68% yield), LCMS (ESI + )m / z:298.4[M+H] + .

[0410] Step 3: Synthesis of compound 26-6:

[0411] Substrate 26-5 (240 mg, 807.13 μmol) and methanol (10 mL) were added to a dry single-mouth bottle, stirred evenly, Pd / C (30 mg) was added, hydrogen was replaced five times, the reaction was stirred at room temperature, and LC-MS was monitored. After the reaction was completed, Pd / C was removed with diatomaceous earth, washed with methanol, and the organic phase was concentrated under reduced pressure to obtain product 26-6 (160 mg, 598.43 μmol, 74.14% yield), LCMS (ESI + )m / z:268.4[M+H] + .

[0412] Step 4: Synthesis of compound 26:

[0413] Add substrate IM-1 (47.5 mg, 132.89 μmol) and tetrahydrofuran (0.5 mL) to a dry single-mouth bottle, stir and dissolve, then add m-CPBA (54 mg, 265.79 μmol), react at room temperature for 1 h, and monitor by LC-MS. After the reaction is completed, add DIPEA (85.88 mg, 664.47 μmol, 115.74 μL), stir for 10 min, and then add compound 26-6 (21.70 mg, 99.88 μmol), and monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure, and the residue is purified by medium pressure liquid chromatography (alkali method) to obtain compound 26 (1.5 mg, 2.35 μmol, 1.77% yield). 1 H NMR (600 MHz, DMSO-d 6 )δ10.37(s,1H),8.90(s,1H),8.11(t,J=8.2Hz,1H),7.80(dd,J=8.3,6.2Hz,2H),7.63(d,J=7.7H z,1H),7.58(d,J=8.4Hz,2H),7.54(d,J=8.5Hz,2H),7.01(d,J=8.8Hz,2H),5.68(ddt,J=16.5,10 .3,5.9Hz,1H),5.35(s,1H),5.01(d,J=10.2Hz,1H),4.88–4.79(m,1H),4.70(d,J=6.0Hz,2H),3. 18(t,J=5.0Hz,4H),2.47(t,J=5.0Hz,4H),2.23(s,3H),1.47(s,6H).LCMS(ESI)m / z:577.3[M+H] + ,HPLC method B:R T =9.20min,purity:97.8%.

[0414] Example 27: Synthesis of Compound 27

[0415]

[0416] Step 1: Synthesis of compound 27-2:

[0417] Substrate 27-1 (400 mg, 2.02 mmol) and DMSO (5 mL) were added to a dry single-mouth bottle, stirred and dissolved, potassium carbonate (836.50 mg, 6.05 mmol) and p-fluoronitrobenzene (313.14 mg, 2.22 mmol) were added, and the mixture was heated to 65°C for 10 hours and monitored by LC-MS. After the reaction was completed, water and ethyl acetate were added for extraction three times, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by medium pressure liquid chromatography to obtain product 27-2 (633 mg, 1.98 mmol, 98.24% yield), LCMS (ESI + )m / z:320.4[M+H] + .

[0418] Step 2: Synthesis of compound 27-3:

[0419] Substrate 27-2 (633 mg, 1.98 mmol) and methanol (5 mL) were added to a dry single-mouth bottle, stirred evenly, Pd / C (60 mg) was added, hydrogen was replaced five times, the reaction was stirred at room temperature, and LC-MS was monitored. After the reaction was completed, Pd / C was removed with diatomaceous earth, washed with methanol, and the organic phase was concentrated under reduced pressure to obtain product 27-3 (459 mg, crude), LCMS (ESI + )m / z:290.4[M+H] + Step 3: Synthesis of compound 27-4:

[0420] Substrate 27-3 (150 mg, 518.36 μmol) and THF (5 mL) were added to a dry single-necked bottle. After stirring to dissolve, LiAlH 4 (19.67 mg, 518.36 μmol), heated to 65 ° C for 4 hours, LC-MS monitoring. After the reaction, 10% sodium hydroxide aqueous solution was added under ice bath to quench, filtered, the filtrate was concentrated under reduced pressure and purified by medium pressure liquid chromatography to obtain product 27-4 (36.59 mg, 180.00 μmol, 34.73% yield), LCMS (ESI + )m / z:204.3[M+H] + .

[0421] Step 4: Synthesis of compound 27:

[0422] Substrate IM-1 (53.61 mg, 0.15 mmol) and tetrahydrofuran (0.5 mL) were added to a dry single-mouth bottle, stirred and dissolved, and then m-CPBA (38.83 mg, 225.00 μmol) was added. The reaction was allowed to react at room temperature for 1 h and monitored by LC-MS. After the reaction was completed, DIPEA (96.93 mg, 750.00 μmol, 130.63 μL) was added, stirred for 10 min, and then compound 27-4 (36.59 mg, 180.00 μmol) was added and monitored by LC-MS. After the reaction was completed, the mixture was concentrated under reduced pressure, and the residue was purified by medium pressure liquid chromatography (alkali method) to obtain compound 27 (18.9 mg, 33.81 μmol, 22.54% yield). 1 H NMR (600 MHz, DMSO-d 6 )δ10.10(s,1H),8.80(s,1H),8.03(s,1H),7.77(s,1H),7.60(d,J=7.7Hz,3H),6.72 (d,J=8.5Hz,2H),5.70–5.61(m,1H),5.32(s,1H),4.99(dd,J=10.3,1.6Hz,1H),4.83 (dd,J=17.1,1.7Hz,1H),4.68(d,J=6.1Hz,2H),3.64(s,2H),3.46(d,J=10.9Hz,2H), 2.51(s,3H),2.05–1.98(m,2H),1.57(s,1H),1.46(s,6H).LCMS(E+)m / z:513.2[M+H] + ,HPLC Method BR T =11.32min,purity:92.6%.

[0423] Example 28: Synthesis of Compound 28

[0424]

[0425] Step 1: Synthesis of compound 28-2:

[0426] Substrate 28-1 (332 mg, 1.56 mmol) and DMSO (5 mL) were added to a dry single-mouth bottle, stirred and dissolved, potassium carbonate (648.42 mg, 4.69 mmol) and p-fluoronitrobenzene (242.73 mg, 1.72 mmol) were added, and the mixture was heated to 80°C for 12 hours and monitored by LC-MS. After the reaction was completed, water and ethyl acetate were added for extraction three times, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by medium pressure liquid chromatography to obtain product 28-2 (512 mg, 1.54 mmol, 98.20% yield), LCMS (ESI + )m / z:334.4[M+H] + .

[0427] Step 2: Synthesis of compound 28-3:

[0428] Substrate 28-2 (640 mg, 1.92 mmol) and methanol (5 mL) were added to a dry single-mouth bottle, stirred evenly, Pd / C (60 mg) was added, hydrogen was replaced five times, the reaction was stirred at room temperature, and LC-MS was monitored. After the reaction was completed, Pd / C was removed with diatomaceous earth, washed with methanol, and the organic phase was concentrated under reduced pressure to obtain product 28-3 (570 mg, crude), LCMS (ESI + )m / z:304.2[M+H] + Step 3: Synthesis of compound 28-4:

[0429] Substrate 28-3 (200 mg, 659.20 μmol) and THF (5 mL) were added to a dry single-necked bottle. After stirring to dissolve, LiAlH 4 (125.08 mg, 3.30 mmol), heated to 65 ° C for 4 hours, LC-MS monitoring. After the reaction, 10% sodium hydroxide aqueous solution was added under ice bath to quench, filtered, the filtrate was concentrated under reduced pressure and purified by medium pressure liquid chromatography to obtain product 28-4 (89 mg, 409.55 μmol, 62.13% yield), LCMS (ESI + )m / z:218.2[M+H] + .

[0430] Step 4: Synthesis of compound 28:

[0431] Add substrate IM-1 (53.61 mg, 0.15 mmol) and tetrahydrofuran (1 mL) to a dry single-mouth bottle, stir and dissolve, then add m-CPBA (38.83 mg, 225.00 μmol), react at room temperature for 1 h, and monitor by LC-MS. After the reaction is completed, add DIPEA (96.93 mg, 750.00 μmol, 130.63 μL), stir for 10 min, and then add compound 28-4 (32.60 mg, 150.00 μmol), and monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure, and the residue is purified by medium pressure liquid chromatography (alkali method) to obtain compound 28 (26.8 mg, 48.90 μmol, 32.60% yield). 1 H NMR (600 MHz, DMSO-d 6 )δ10.11(s,1H),8.81(s,1H),8.04(s,1H),7.75(s,1H),7.65–7.43(m,3H),6.79(d,J=8.6Hz,2H ),5.66(ddt,J=16.4,10.2,6.0Hz,1H),5.32(s,1H),4.99(dd,J=10.3,1.4Hz,1H),4.82(dd,J=17 .1,1.5Hz,1H),4.68(d,J=5.9Hz,2H),3.35(d,J=10.5Hz,2H),3.24(s,2H),2.81(d,J=10.3Hz,2H ),2.25(s,3H),2.00–1.94(m,2H),1.66(d,J=7.6Hz,2H),1.46(s,6H).LCMS(E+)m / z:527.4[M+H] + ,HPLC Method BR T =10.46min,purity:96.1%.

[0432] Example 29: Synthesis of Compound 29

[0433]

[0434] Step 1: Synthesis of compound 29-2

[0435] Substrate 29-1 (233 mg, 891.63 μmol) and methanol (10 mL) were added to a dry single-mouth bottle, stirred and dissolved, palladium carbon (10 mg) was added, hydrogen was replaced five times, reacted at room temperature for 2 hours, and monitored by LC-MS. After the reaction was completed, it was filtered with diatomaceous earth, washed with methanol, and the organic phase was concentrated by vacuum distillation to obtain compound 29-2 (200 mg, 866 μmol), LCMS (ESI) m / z: 232.2 [M+H] + .

[0436] Step 2: Synthesis of compound 29

[0437] In a dry single-mouth bottle, add substrate IM-1 (38.63 mg, 108.07 μmol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (30 mg, 162.10 μmol) and react at room temperature for one hour. Then add N,N-diisopropylethylamine (20.95 mg, 162.10 μmol) and 29-2 (25 mg, 0.18 mmol) to the reaction solution and react at room temperature overnight. LC-MS monitoring, after the reaction is completed, cool the reaction solution to room temperature, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 29 (18 mg, 33.29 μmol). LCMS (ESI) m / z: 541.4 [M+H] + , HPLC Method BR T =5.14min,purity>90.0%. 1 H NMR (600 MHz, Methanol-d 4 )δ8.76(s,1H),7.95–7.93(m,1H),7.76–7.74(m,1H),7.62(d,J=7.8Hz,1H),7.44(s,2H) ,6.70–6.62(m,2H),5.72–5.68(m,1H),5.03(d,J=10.2Hz,1H),4.95–4.88(m,2H),4.80(d ,J=6.0Hz,2H),4.00(d,J=8.6Hz,1H),3.94–3.89(m,1H),3.53–3.45(m,4H),3.35(s,3H), 3.22–3.18(m,1H),2.41–2.33(m,2H),2.16–2.12(m,2H),1.98–1.95(m,1H),1.57(s,6H).

[0438] Example 30: Synthesis of Compound 30

[0439]

[0440] Step 1: Synthesis of compound 30-2

[0441] Add substrate 30-1 (200 mg, 912.24 μmol) to a dry single-mouth bottle and dissolve it in tetrahydrofuran (3 mL), add triethylamine (461 mg, 4.56 mmol) and trifluoroethyl trifluoromethanesulfonate (317 mg, 1.37 mmol). Raise the room temperature to 80 ° C and stir overnight. Monitor by LC-MS. After the reaction is completed, distill and concentrate under reduced pressure to obtain compound 30-2 (235 mg, 780 μmol), LCMS (ESI) m / z: 302.1 [M+H] + .

[0442] Step 2: Synthesis of compound 30-3

[0443] Substrate 30-2 (235 mg, 780 μmol) and methanol (3 mL) were added to a dry single-mouth bottle. After stirring and dissolving, palladium carbon (25 mg) was added, and hydrogen was replaced five times. The reaction was carried out at room temperature for 2 hours and monitored by LC-MS. After the reaction was completed, it was filtered with diatomaceous earth, washed with methanol, and the organic phase was concentrated by vacuum distillation to obtain compound 30-3 (183 mg, 675 μmol). LCMS (ESI) m / z: 272.2 [M+H] + .

[0444] Step 3: Synthesis of compound 30

[0445] In a dry single-mouth bottle, add substrate IM-1 (27 mg, 75.54 μmol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (23 mg, 113.31 μmol) and react at room temperature for one hour. Then add N,N-diisopropylethylamine (48.81 mg, 377.1 μmol) and 30-3 (30.7 mg, 113.31 μmol) to the reaction solution and react at room temperature overnight. LC-MS monitoring, after the reaction is completed, the reaction solution is cooled to room temperature, concentrated under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (alkaline) to obtain compound 30 (10.7 mg, 18.43 μmol). LCMS (ESI) m / z: 581.3 [M + H] + , HPLC Method B Rt=8.07min, purity>89.7%. 1 H NMR (600 MHz, DMSO-d 6)δ10.11(s,1H),8.80(s,1H),7.99-7.96(m,1H),7.75(s,1H),7.62(d,J=7.8Hz,1H),7. 47(s,2H),6.62(d,J=8.4Hz,2H),6.19(d,J=23.4Hz,1H),5.69-5.63(m,1H),4.99(d,J=1 0.2Hz,1H),4.83(d,J=17.4Hz,1H),4.67(s,2H),3.89(s,2H),3.51(s,2H),3.29(s,3H) ,2.97(s,3H),2.22(s,2H),2.00(d,J=7.8Hz,2H),1.76(d,J=14.4Hz,1H),1.46(s,6H).D 2 O: 1 H NMR (600 MHz, DMSO-d 6 )δ8.81(s,1H),8.01-7.98(m,1H),7.74(s,1H),7.62(d,J=7.8Hz,1H),7.49(s, 2H),6.64(d,J=8.4Hz,2H),5.69-5.63(m,1H),5.02(d,J=10.2Hz,1H),4.83(d, J=17.4Hz,1H),4.68(s,2H),3.90-3.88(m,2H),3.43-3.42(m,4H),3.14–3.12( m,1H),2.97(s,3H),2.23–2.20(m,2H),2.03(s,2H),1.82(s,1H),1.47(s,6H).

[0446] Example 31: Synthesis of Compound 31

[0447]

[0448] Step 1: Synthesis of compound 31-2

[0449] Substrate 31-1 (2.0 g, 7.80 mmol) and methanol (10 mL) were added to a dry single-mouth bottle, stirred and dissolved, palladium carbon (200 mg) was added, hydrogen was replaced five times, reacted at room temperature for 2 hours, and monitored by LC-MS. After the reaction was completed, it was filtered with diatomaceous earth, washed with methanol, and the organic phase was concentrated by vacuum distillation to obtain compound 31-2 (1.6 g, 7.07 mmol), LCMS (ESI) m / z: 232.2 [M+H] + .

[0450] Step 2: Synthesis of compound 31

[0451] In a dry single-mouth bottle, add substrate IM-1 (100 mg, 279.78 μmol) and tetrahydrofuran (1.5 mL) to dissolve. Add m-chloroperbenzoic acid (120 mg, 559.55 μmol) and react at room temperature for one hour. Then add N, N-diisopropylethylamine (180.79 mg, 1.40 mmol) and 31-2 (126.61 mg, 559.55 μmol) to the reaction solution and react at room temperature overnight. LC-MS monitoring, after the reaction is completed, the reaction solution is cooled to room temperature, concentrated under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (alkaline) to obtain compound 31 (40 mg, 74.68 μmol). LCMS (ESI) m / z: 536.2 [M+H] + ,HPLC method B:R T =8.58min,purity:>96.1%. 1 H NMR (600 MHz, DMSO-d 6 )δ10.33(s,1H),8.89(s,1H),8.04–8.02(m,1H),7.80–7.71(m,3H),7.61(d,J=7. 8Hz,1H),7.23(d,J=8.4Hz,2H),6.81(d,J=7.4Hz,1H),6.73–6.71(m,1H),6.70–6. 67(m,2H),5.68–5.64(m,1H),5.32(s,1H),5.00(d,J=10.2Hz,1H),4.83(d,J=17. 4Hz,1H),4.69(d,J=6.0Hz,2H),4.27–4.26(m,2H),3.68–3.66(m,2H),1.46(s,7H)

[0452] Example 32: Synthesis of Compound 32

[0453]

[0454] Step 1: Synthesis of compound 32-1

[0455] In a dry single-mouth bottle, 4-1 (800 mg, 6.39 mmol) was dissolved in ethanol (10 mL), and potassium hydroxide (717 mg, 12.78 mmol) and p-nitrobenzaldehyde (1.16 g, 7.67 mmol) were added in sequence. The reaction mixture was stirred at room temperature overnight. After the reaction was completed, the mixture was concentrated under reduced pressure by LC-MS monitoring, and the residue was purified by MPLC to obtain compound 32-1 (863 mg, 3.34 mmol). LCMS (ESI) m / z: 259.1 [M+H] + .

[0456] Step 2: Synthesis of compound 32-2

[0457] Add trimethylsulfoxide iodide (2.94 g, 13.37 mmol) and sodium hydroxide (1.07 g, 26.73 mmol) to a single-mouth bottle and dissolve in tetrahydrofuran (20 mL), replace nitrogen, and react at room temperature for 1 hour. Add 32-1 (863 mg, 3.34 mmol), replace nitrogen again, stir at 65 ° C for 1 hour, and monitor the reaction by LCMS. After the reaction is completed, concentrate under reduced pressure to remove the solvent, add ethyl acetate and water to extract 3 times, dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is purified by column chromatography to obtain compound 32-2 (493 mg, 1.81 mmol). LCMS (ESI) m / z: 273.1 [M+H] + .

[0458] Step 3: Synthesis of compound 32-3

[0459] Substrate 32-2 (493 mg, 1.81 mmol) and methanol (5 mL) were added to a dry single-mouth bottle, stirred and dissolved, palladium carbon (50 mg) was added, hydrogen was replaced five times, reacted at room temperature for 2 hours, and monitored by LC-MS. After the reaction was completed, it was filtered with diatomaceous earth, washed with methanol, and the organic phase was concentrated by vacuum distillation to obtain compound 32-3 (395 mg, 1.63 mmol), LCMS (ESI) m / z: 243.2 [M+H] + .

[0460] Step 4: Synthesis of compound 32

[0461] In a dry single-mouth bottle, add substrate IM-1 (36 mg, 100.72 μmol) and tetrahydrofuran (1.5 mL) to dissolve. Add m-chloroperbenzoic acid (26 mg, 151 μmol) and react at room temperature for one hour. Then add N, N-diisopropylethylamine (65 mg, 504 μmol) and 32-3 (48.8 mg, 201 μmol) to the reaction solution and react at room temperature overnight. LC-MS monitoring, after the reaction is completed, the reaction solution is cooled to room temperature, concentrated under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (alkaline) to obtain compound 32 (7.2 mg, 13.05 μmol). LCMS (ESI) m / z: 552.3 [M+H] + ,HPLC method A:R T =9.90min,purity:>99.1%. 1 H NMR (600 MHz, DMSO-d 6)δ10.26(s,1H),8.88(s,1H),8.04–8.02(m 1H),7.75(d,J=8.4Hz,1H),7.62(d,J=8.4Hz,3H),7.26(d,J=8.4Hz,2H),5.67–5.65(m,1H),5.34(s ,1H),5.00(d,J=10.2Hz,1H),4.83(d,J=17.4Hz,1H),4.69(d,J=6.0Hz,2H),2.96–2.93(m,2H),2.7 7–2.70(m,1H),2.49(d,J=12.6Hz,1H),2.45–2.43(m,1H),2.21–2.20(m,1H),2.01-1.97(m,1H),1. 92–1.89(m,2H),1.87–1.79(m,J=2.6Hz,1H),1.67–1.62(m,1H),1.59(d,J=9.6Hz,1H),1.46(s,6H).

[0462] Example 33: Synthesis of Compound 33

[0463]

[0464] Step 1: Synthesis of compound 33-2

[0465] In a dry single-mouth bottle, 23-5 (100 mg, 404.38 μmol) was dissolved in tetrahydrofuran (5 ml), and triethylamine (409.19 mg, 4.04 mmol) and 33-1 (140.79 mg, 606.57 μmol) were added in sequence. The reaction mixture was stirred at 80°C for 3 hours. After the reaction was completed, it was concentrated under reduced pressure, and the residue was purified by MPLC to obtain compound 33-2 (70 mg, 212.56 μmol). LCMS (ESI) m / z: 330.1 [M+H] + .

[0466] Step 2: Synthesis of compound 33-3

[0467] 33-2 (80 mg, 0.24 mmol) was added to a 50 mL round-bottom flask and dissolved in methanol (3 mL). A catalytic amount of palladium carbon (10 mg) was added and stirred at room temperature for 2 hours under a hydrogen atmosphere. LC-MS monitoring. After the reaction was completed, the reaction solution was filtered through diatomaceous earth, washed twice with methanol, and the filtrate was concentrated under reduced pressure to obtain compound 33-3 (20 mg, 0.17 mmol,). LCMS (ESI) m / z: 300.2 [M+H] + .

[0468] Step 3: Synthesis of compound 33

[0469] Add substrate IM-6 (48.55 mg, 130.00 μmol) and tetrahydrofuran (3 mL) to a dry single-mouth bottle, stir and dissolve, then add N,N-diisopropylethylamine (84.01 mg, 650.00 μmol) and 33-3 (44 mg, 146.99 μmol), and react at room temperature for 2 hours. LC-MS monitoring. After the reaction is completed, the solvent is removed by distillation under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (basic) to obtain compound 33 (20 mg, 31.77 μmol). LCMS (ESI) m / z: 609.2 [M+H] + ,HPLC method A:R T =5.8min,purity:96.7%. 1 H NMR (600 MHz, DMSO-d 6 )δ10.08(s,1H),8.80(s,1H),8.01-8.00(m,1H),7.77(s,1H),7.60(d,J=7.8Hz ,1H),7.54–7.40(m,2H),6.75(d,J=8.4Hz,2H),5.67-5.63(m,1H),5.37(s,1H), 5.00(d,J=10.2Hz,1H),4.83(d,J=16.8Hz,1H),4.69(s,2H),3.47(s,2H),3.08( s,4H),2.95(s,2H),2.70(s,2H),2.07(s,2H),1.66-1.59(m,2H),1.47(s,6H).D 2 O: 1 H NMR (600 MHz, DMSO-d 6 )δ8.81(s,1H),8.02-8.01(m,1H),7.75(s,1H),7.61(d,J=7.2Hz,1H),7.54(s,2 H),6.75(d,J=7.8Hz,2H),5.68-5.64(m,1H),5.02(d,J=10.2Hz,1H),4.84(d,J=1 6.8Hz,1H),4.70(s,2H),3.52(d,J=10.8Hz,2H),3.09–3.04(m,4H),2.94(d,J=1 0.2Hz, 2H), 2.69 (d, J = 10.2Hz, 2H), 2.08 (s, 2H), 1.67-1.58 (m, 2H), 1.48 (s, 6H).

[0470] Example 34: Synthesis of Compound 34

[0471]

[0472] Step 1: Synthesis of compound 34-1

[0473] Add substrate 23-5 (100 mg, 404.38 μmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (3 mL), add 2-bromoethyl methyl ether (84 mg, 606 μmol), potassium carbonate (111.77 mg, 808.76 μmol) and potassium iodide (13.43 mg, 80.88 μmol), and react at 85°C for 3 hours. LC-MS monitoring. After the reaction is completed and the reaction solution is cooled to room temperature, water is added, and ethyl acetate is extracted 3 times. The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by column chromatography to obtain compound 34-1 (65 mg, 212.86 μmol). LCMS (ESI) m / z: 306.2 [M+H] + .

[0474] Step 2: Synthesis of compound 34-2

[0475] Add 34-1 (80 mg, 261.98 μmol) to a 50 mL round-bottom flask and dissolve in methanol (3 mL). Add a catalytic amount of palladium carbon (10 mg) and stir at room temperature for 2 hours under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 34-2 (29 mg, 0.10 mmol). LCMS (ESI) m / z: 276.2 [M+H] + .

[0476] Step 3: Synthesis of compound 34

[0477] Add substrate IM-6 (37 mg, 99.08 μmol) and tetrahydrofuran (3 mL) to a dry single-mouth bottle, stir and dissolve, then add N,N-diisopropylethylamine (64.03 mg, 495.41 μmol) and 34-2 (28.65 mg, 104.04 μmol), and react at room temperature for 2 hours. LC-MS monitoring. After the reaction is completed, the solvent is removed by distillation under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (alkaline) to obtain compound 34 (10 mg, 17.10 μmol). LCMS (ESI) m / z: 585.3 [M+H] + ,HPLC method A:R T =5.4min,purity:96.4%. 1 H NMR (600 MHz, DMSO-d 6)δ10.09(s,1H),8.80(s,1H),8.03(s,1H),7.76(d,J=7.2Hz,1H),7.61–7.56(m,3H ),6.75(s,2H),5.69-5.63(m,1H),5.32(s,1H),4.99(d,J=10.2Hz,1H),4.83(d,J=1 7.4,1H),4.68(d,J=4.2Hz,2H),3.54(d,J=10.2Hz,2H),3.40(s,4H),3.18(s,3H),3 .06-2.98(m,4H),2.44-2.29(m,2H),2.13(s,2H),1.68–1.57(m,2H),1.46(s,6H).D 2 O: 1 H NMR (600 MHz, DMSO-d 6 )δ8.81(s,1H),8.03(s,1H),7.76-7.75(m,1H),7.61(d,J=7.8Hz,1H)7.56–7.50(m,2 H),6.77(s,2H),5.69-5.63(m,1H),5.01(d,J=10.2Hz,1H),4.83(d,J=16.8Hz,,1H), 4.68(d,J=4.8Hz,2H),3.63(s,2H),3.55(d,J=10.8Hz,2H),3.41(s,2H),3.18(s,3H) ,3.05-2.96(m,4H),2.45-2.36(m,2H),2.14(s,2H),1.66-1.63(m,2H),1.47(s,6H).

[0478] Example 35: Synthesis of Compound 35

[0479]

[0480] Step 1: Synthesis of compound 35-2

[0481] In a 100mL dry round-bottom flask, add 35-1 (2.59g, 11mmol) under nitrogen protection, and add tetrahydrofuran (25mL) to dissolve. The temperature dropped to -78℃, and n-LiBu (5.5mL, 2M in THF, 11mmol) was slowly added dropwise into the reaction system. After stirring at -78℃ for 30 minutes, N-methyl-4-piperidone (1.24g, 11.00mmol) was added. Slowly warm to room temperature and stir for 1 hour. Add saturated NH 4The reaction was quenched with Cl solution and extracted with ethyl acetate three times. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and the residue was purified by column chromatography to obtain compound 35-2 (2.63 g, 9.73 mmol). LCMS (ESI) m / z: 270.1 [M+H] + Step 2: Synthesis of compound 35-3

[0482] In a dry microwave tube, under nitrogen protection, add substrate 35-2 (675.4 mg, 2.5 mmol), ammonia (1.75 g, 50.00 mmol), cuprous oxide (17.9 mg, 0.125 mmol), sodium iodide (242.66 mg, 1.62 mmol), potassium carbonate (69.10 mg, 0.5 mmol) and N, N'-dimethylethylenediamine (22 mg, 0.25 mmol), and then add ethylene glycol (5 mL). Under nitrogen environment, heat the reaction at 110 ° C for 12 hours. LC-MS monitoring, after the reaction is completed, the reaction system is cooled to room temperature, filtered, extracted with ethyl acetate 3 times, the organic phase is dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue is purified by column chromatography to obtain compound 35-3 (346 mg, 1.68 mmol). LCMS (ESI) m / z: 207.2 [M+H] + .

[0483] Step 3: Synthesis of compound 35

[0484] In a dry single-mouth bottle, add substrate IM-1 (71.5 mg, 0.2 mmol), add tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (51.8 mg, 0.3 mmol), react at room temperature for 30 minutes, then add N, N-diisopropylethylamine (129.2 mg, 1.00 mmol) and 35-3 (61.9 mg, 0.3 mmol) to the reaction solution, and react at room temperature for 4 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 35 (16 mg, 31.03 μmol). 1 HNMR (600 MHz, DMSO-d 6)δ10.27(s,1H),8.88(s,1H),8.06-8.04(m,1H),7.76(d,J=8.4Hz,1H),7.65(d,J=26.4Hz,3 H),7.43(d,J=8.4Hz,2H),5.67–5.64(m,1H),5.34(s,1H),5.00(d,J=10.2Hz,1H),4.83(d,J =17.4Hz,1H),4.73(s,1H),4.68(d,J=6.0Hz,2H),2.58(d,J=10.8Hz,2H),2.40–2.38(m,2H) ,2.23(s,3H),2.01–1.91(m,2H),1.61–1.55(m,2H),1.46(s,6H).LCMS(ESI)m / z:516.3[M+H] + , HPLC R T =4.68min,purity>92.7%.

[0485] Example 36: Synthesis of Compound 36

[0486]

[0487] Step 1: Synthesis of compound 36-2

[0488] Add 36-1 (5 g, 44.22 mmol) to a dry single-mouth bottle and dissolve it in dichloromethane (50 mL). Add triethylamine (8.95 g, 88.44 mmol, 12.33 mL), 4-dimethylaminopyridine (540.22 mg, 4.42 mmol) and di-tert-butyl dicarbonate (10.62 g, 48.64 mmol) in sequence. The reaction mixture was stirred at room temperature for 2 hours. LC-MS monitoring. After the reaction was completed, it was diluted with water and extracted with dichloromethane three times. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound 36-2 (4.62 g, 21.67 mmol). LCMS (ESI) m / z: 214.1 [M+H] + .

[0489] Step 2: Synthesis of compound 36-3

[0490] Add 36-2 (4.62 g, 21.67 mmol) to a 50 mL round-bottom flask and dissolve in ethanol (3 mL). Add a catalytic amount of palladium carbon (400 mg) and stir at room temperature for 6 hours under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with ethanol, and concentrate the filtrate under reduced pressure to obtain compound 36-3 (3.85 g, 21.01 mmol). LCMS (ESI) m / z: 184.1 [M+H]+ .

[0491] Step 3: Synthesis of compound 36-4

[0492] Add substrate 36-3 (3.85 g, 21.01 mmol) dissolved in methanol (40 mL) to a dry single-mouth bottle, add formaldehyde (1.26 g, 42.03 mmol) and acetic acid (2 mL), and stir at room temperature for 30 minutes. Add sodium cyanoborohydride (3.30 g, 52.54 mmol), raise the temperature to 50 ° C and react for 12 hours. After the reaction is completed, dilute with water, add saturated sodium bicarbonate to adjust to alkalinity, and extract with dichloromethane three times. The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by column chromatography to obtain compound 36-4 (4.01 g, 18.98 mmol). LCMS (ESI) m / z: 212.1 [M+H] + .

[0493] Step 4: Synthesis of compound 36-5

[0494] Add 36-4 (4.01 g, 18.98 mmol) and dissolve it in dichloromethane (40 ml) in a dry single-mouth bottle. Add trifluoroacetic acid (21.64 g, 189.81 mmol, 14.53 mL) and react at room temperature for 30 minutes. LC-MS monitoring, after the reaction is completed, remove the solvent by vacuum distillation to obtain compound 36-5 (1.86 g, 16.73 mmol). LCMS (ESI) m / z: 112.1 [M+H] + .

[0495] Step 5: Synthesis of compound 36-6

[0496] In a dry single-mouth bottle, 36-5 (1.86 g, 16.73 mmol) was dissolved in dimethyl sulfoxide (15 ml), and p-fluoronitrobenzene (2.36 g, 16.73 mmol) and potassium carbonate (5.78 g, 41.84 mmol) were added in sequence. The reaction mixture was stirred at 100 °C overnight. LC-MS monitoring, after the reaction was completed, the mixture was diluted with water, extracted with ethyl acetate three times, the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and the residue was purified by column chromatography to obtain compound 36-6 (3.56 g, 15.33 mmol). LCMS (ESI) m / z: 233.1 [M+H] + .

[0497] Step 6: Synthesis of compound 36-7

[0498] Add 36-6 (3.56 g, 15.33 mmol) to a dry single-mouth bottle and dissolve it in ethanol (35 mL). Add a catalytic amount of palladium carbon (350 mg) and stir at room temperature for 3 hours under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with ethanol, and concentrate the filtrate under reduced pressure to obtain compound 36-7 (2.95 g, 14.59 mmol). LCMS (ESI) m / z: 203.1 [M+H] + .

[0499] Step 7: Synthesis of compound 36

[0500] In a dry single-mouth bottle, add substrate IM-1 (45.51 mg, 225.00 μmol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (38.83 mg, 225.00 μmol) and react at room temperature for 30 minutes. Then add N,N-diisopropylethylamine (96.93 mg, 750.00 μmol) and 36-7 (53.61 mg, 0.15 mmol) to the reaction solution and react at room temperature for 4 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 36 (9 mg, 17.59 μmol). LCMS (ESI) m / z: 512.2 [M+H] + , HPLC Method BR T =7.26min,purity>98.0%. 1 HNMR (600 MHz, DMSO-d 6 )δ10.39(s,1H),8.91(s,1H),8.09(s,1H),7.87(s,1H),7.82(d,J=8.4H z,2H),7.78(d,J=8.4Hz,1H),7.72(d,J=9.0Hz,2H),7.64(d,J=7.8Hz,1H ),7.45(s,1H),5.71-5.66(m,1H),5.34(s,1H),5.00(d,J=10.2Hz,1H), 4.84(d,J=17.4Hz,1H), 4.70(d,J=6.0Hz,2H), 2.70(s,6H), 1.47(s,6H).

[0501] Example 37: Synthesis of Compound 37

[0502]

[0503] Step 1: Synthesis of compound 37-1

[0504] Add substrate 23-5 (150 mg, 606.57 μmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (3 mL), add 1,2-fluorobromoethane (84 mg, 661.65 μmol), potassium carbonate (165 mg, 1.19 mmol) and potassium iodide (20 mg, 120.5 μmol), and react at 90°C for 3 hours. LC-MS monitoring. After the reaction is completed, the reaction solution is cooled to room temperature, water is added, filtered, and washed with methanol. The organic phase is concentrated under reduced pressure to obtain compound 37-1 (80 mg, 272.7 μmol). LCMS (ESI) m / z: 394.2 [M+H] + .

[0505] Step 2: Synthesis of compound 37-2

[0506] Add 37-1 (80 mg, 272.7 μmol) to a 50 mL round-bottom flask and dissolve in methanol (3 mL). Add a catalytic amount of palladium carbon (10 mg) and stir at room temperature for 2 hours under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 37-2 (50 mg, 0.19 mmol). LCMS (ESI) m / z: 264.2 [M+H] + .

[0507] Step 3: Synthesis of compound 37

[0508] Add substrate IM-6 (42 mg, 112.47 μmol) and tetrahydrofuran (3 mL) to a dry single-mouth bottle, stir and dissolve, then add N,N-diisopropylethylamine (72.68 mg, 562.36 μmol) and 37-2 (30 mg, 113.92 μmol), and react at room temperature for 2 hours. LC-MS monitoring. After the reaction is completed, the solvent is removed by distillation under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (alkaline) to obtain compound 37 (25 mg, 42.65 μmol). LCMS (ESI) m / z: 585.3 [M+H] + ,HPLC method A:R T =5.4min,purity:96.4%. 1H NMR(600MHz,Chloroform-d)δ8.74(s,1H),7.82(s,1H),7.71(d,J=7.8Hz,1H),7.36(s,2H) ,7.26(d,J=7.2Hz,1H),6.76(s,2H),5.67-5.60(m,1H),4.97(d,J=10.2Hz,1H),4.87(d,J= 17.4Hz,1H),4.67(d,J=6.0Hz,2H),4.55-4.74(m,2H),3.94(brs,1H),3.57-3.55(m,2H),3 .10-2.93(m,4H),2.70-2.51(m,4H),2.12(s,2H),1.67(s,2H),1.51(s,6H),1.25(brs,1H).

[0509] Example 38: Synthesis of Compound 38

[0510]

[0511] Step 1: Synthesis of compound 38-1

[0512] IM-4 (500 mg, 1.57 mmol) was added to a dry single-mouth bottle and dissolved in dichloromethane (3 ml). Trifluoroacetic acid (1 ml) was added and the reaction was allowed to react at room temperature for 1 hour. After the reaction was completed, the solvent was removed by distillation under reduced pressure to obtain compound 38-1 (330 mg, 1.51 mmol). LCMS (ESI) m / z: 220.1 [M+H] + .

[0513] Step 2: Synthesis of compound 38-2

[0514] Add substrate 38-1 (100 mg, 456.12 μmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (2 mL), add 2-bromoethanol (57.00 mg, 456.12 μmol), potassium carbonate (94.55 mg, 684.18 μmol) and potassium iodide (15.14 mg, 91.22 μmol), and react at 90°C for 2 hours. LC-MS monitoring. After the reaction is completed, the reaction solution is cooled to room temperature, water is added, filtered, and washed with methanol. The organic phase is concentrated under reduced pressure to obtain compound 38-2 (93 mg, 353 μmol). LCMS (ESI) m / z: 264.1 [M+H] + .

[0515] Step 3: Synthesis of compound 38-3

[0516] Under nitrogen protection, add substrate 38-2 (93 mg, 353.22 μmol) to a dry single-mouth bottle, dissolve in tetrahydrofuran (2.5 mL), and add triethylamine (142.97 mg, 1.41 mmol). The system temperature drops to 0°C, and acetyl chloride (41.59 mg, 529.83 μmol) is added. Slowly warm to room temperature and stir for 1 hour. LC-MS monitoring. After the reaction is completed, concentrate under reduced pressure to obtain compound 38-3 (96 mg, 314.8 μmol). LCMS (ESI) m / z: 292.1 [M+H] + .

[0517] Step 4: Synthesis of compound 38-4

[0518] Add 38-3 (277 mg, 907.22 μmol) to a 50 mL round-bottom flask and dissolve in methanol (3 mL). Add a catalytic amount of palladium carbon (30 mg) and stir at room temperature for 2 hours under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 38-4 (177 mg, 642 μmol). LCMS (ESI) m / z: 262.2 [M+H] + .

[0519] Step 5: Synthesis of compound 38-5

[0520] In a dry single-mouth bottle, add substrate IM-1 (40 mg, 111.91 μmol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (38.62 mg, 223.82 μmol) and react at room temperature for 30 minutes. Then add N,N-diisopropylethylamine (72.32 mg, 559.55 μmol) and 38-4 (30.81 mg, 111.91 μmol) to the reaction solution and react at room temperature for 4 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure to obtain crude product 38-5 (300 mg, crude). LCMS (ESI) m / z: 571.3 [M+H] + .

[0521] Step 6: Synthesis of compound 38

[0522] In a dry single-mouth bottle, add substrate 38-5 (300 mg, crude) and methanol (3 mL) to dissolve. Add sodium hydroxide (41.05 mg, 1.03 mmol) and water (0.2 mL) and react at room temperature for 30 minutes. Monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure, dilute with water, and extract with ethyl acetate three times. The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product is purified by preparative high performance liquid chromatography (alkaline) to obtain compound 38 (10.36 mg, 19.09 μmol). LCMS (ESI) m / z: 543.2 [M+H] + .HPLC method A:R T =4.82min,purity:100.0%. 1 H NMR (600 MHz, DMSO-d 6 )δ10.04(s,1H),8.79(s,1H),8.02–7.99(m,1H),7.74(s,1H),7.59(d,J=7.2Hz,1H),7.49(s,2 H),6.55(d,J=7.8Hz,2H),5.67-5.63(m,1H),5.32(s,1H),4.99(d,J=10.2Hz,1H),4.82(d,J=17 .4Hz,1H),4.69-4.67(m,2H),4.37-4.35(m,1H),4.26(s,1H),3.54(s,1H),3.38-3.36(m,3H),3 .14(d,J=9.0Hz,1H),2.87(d,J=9.0Hz,1H),2.48-2.46(m,2H),1.83-1.77(m,2H),1.46(s,6H).

[0523] Example 39: Synthesis of Compound 39

[0524]

[0525] Step 1: Synthesis of compound 39

[0526] In a dry single-mouth bottle, add substrate IM-3 (30.49 mg, 150.00 μmol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (25.88 mg, 150.00 μmol) and react at room temperature for 30 minutes. Then add N, N-diisopropylethylamine (19.39 mg, 150.00 μmol) and compound 27-4 (30.49 mg, 150.00 μmol) to the reaction solution and react at room temperature for 4 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 39 (26 mg, 38.83 μmol). LCMS (ESI) m / z: 546.2 [M+H] + , HPLC Method BR T =5.93min,purity>81.5%. 1 H NMR (600 MHz, DMSO-d 6 )δ10.07(s,1H),8.78(s,1H),7.78(s,1H),7.61(s,2H),7.32(d,J=7.8Hz,1H),6 .71(d,J=7.8Hz,2H),6.59(d,J=8.4Hz,1H),5.68–5.60(m,1H),5.00(d,J=10.2Hz ,1H),4.90(d,J=17.4Hz,1H),4.75(s,2H),3.60(s,2H),3.44(d,J=11.4Hz,2H), 3.38(s,6H),3.35(d,J=8.4Hz,2H),2.44(s,1H),2.00(s,3H),1.55–1.53(m,1H).

[0527] Example 40: Synthesis of Compound 40

[0528]

[0529] Step 1: Synthesis of compound 40-2

[0530] Dissolve 40-1 (0.4 g, 2 mmol) in dimethyl sulfoxide (15 mL) in a dry single-mouth bottle, and add p-fluoronitrobenzene (0.28 g, 2 mmol) and potassium carbonate (1.11 g, 4 mmol) in sequence. The reaction mixture was stirred at 100°C overnight. LC-MS monitoring, after the reaction was completed, dilute with water, extract with ethyl acetate three times, dry the organic phase with anhydrous sodium sulfate and concentrate under reduced pressure, the residue was purified by column chromatography to obtain compound 40-2 (0.63 g, 2.56 mmol). LCMS (ESI) m / z: 248.1 [M+H] + .

[0531] Step 2: Synthesis of compound 40-3

[0532] Under nitrogen protection, add substrate 40-2 (71.4 mg, 300 μmol) to a dry single-mouth bottle, dissolve in tetrahydrofuran (2.5 mL), and add triethylamine (121.4 mg, 1.2 mmol). The system temperature is reduced to 0°C, and acetic anhydride (61.2 mg, 600 μmol) is added. Slowly warm to room temperature and stir for 12 hours. LC-MS monitoring. After the reaction is completed, it is concentrated under reduced pressure, and the residue is purified by MPLC to obtain compound 40-3 (25.3 mg, 87.4 μmol). LCMS (ESI) m / z: 290.2 [M+H] + .

[0533] Step 3: Synthesis of compound 40-4

[0534] 40-3 (173 mg, 598 μmol) was added to a 50 mL round-bottom flask and dissolved in methanol (5 mL). A catalytic amount of palladium carbon (18 mg) was added and stirred at room temperature for 2 hours under a hydrogen atmosphere. LC-MS monitoring. After the reaction was completed, the reaction solution was filtered through diatomaceous earth, washed twice with methanol, and the filtrate was concentrated under reduced pressure to obtain compound 40-4 (122 mg, 470 μmol). LCMS (ESI) m / z: 260.2 [M+H] + .

[0535] Step 4: Synthesis of compound 40

[0536] In a dry single-mouth bottle, add substrate IM-1 (35.7 mg, 100 μmol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (25.85 mg, 150 μmol) and react at room temperature for 30 minutes. Then add N,N-diisopropylethylamine (64.54 mg, 500 μmol) and 40-4 (51.81 mg, 0.2 mmol) to the reaction solution, and raise the temperature to 60°C to react for 4 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 40 (18 mg, 31.65 μmol). LCMS (ESI) m / z: 569.3 [M+H] + , HPLC Method B Rt=6.28min, purity>63.4%. 1 HNMR (600 MHz, DMSO-d 6)δ10.14(s,1H),8.82(s,1H),8.04(s,1H),7.89(s,1H),7.75(s,1H),7.67–7.47(m,3H),6.93(d,J=8. 4Hz,2H),5.68–5.64(m,1H),5.32(s,1H),4.99(d,J=10.2Hz,1H),4.82(d,J=17.4Hz,1H),4.68(s,2H), 4.37–4.35(m,1H),4.13–4.06(m,1H),3.69(s,2H),3.62(d,J=7.8Hz,1H),2.66–2.55(m,3H),2.39(s,1 H),1.79(d,J=13.2Hz,1H),1.59(d,J=13.2Hz,1H),1.46(s,6H),1.45–1.42(m,1H),1.36–1.24(m,2H).

[0537] Example 41: Synthesis of Compound 41

[0538]

[0539] Step 1: Synthesis of compound 41-1

[0540] Add substrate 38-1 (140 mg, 638.57 μmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (3 mL), add 2-iodopropane (131 mg, 770.63 μmol), and potassium carbonate (124 mg, 897.25 μmol), and react at 90°C for 3 hours. LC-MS monitoring. After the reaction is completed, the reaction solution is cooled to room temperature, water is added, and ethyl acetate is extracted 3 times. The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by column chromatography to obtain compound 41-1 (80 mg, 306.14 μmol). LCMS (ESI) m / z: 262.2 [M+H] + .

[0541] Step 2: Synthesis of compound 41-2

[0542] Add 41-1 (80 mg, 306.14 μmol) to a 50 mL round-bottom flask and dissolve in methanol (3 mL). Add a catalytic amount of palladium carbon (20 mg) and stir at room temperature for 2 hours under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 41-2 (60 mg, crude). LCMS (ESI) m / z: 232.2 [M+H] + .

[0543] Step 3: Synthesis of compound 41

[0544] Add substrate IM-6 (45 mg, 120.50 μmol) and tetrahydrofuran (3 mL) to a dry single-mouth bottle, stir and dissolve, then add N,N-diisopropylethylamine (77.87 mg, 602.52 μmol) and 41-2 (30 mg, 129.68 μmol), and react at room temperature for 2 hours. LC-MS monitoring. After the reaction is completed, the solvent is removed by distillation under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (basic) to obtain compound 41 (20 mg, 36.99 μmol). LCMS (ESI) m / z: 541.3 [M+H] + ,HPLC method A:R T =5.1min,purity:95.3%. 1 H NMR (600 MHz, DMSO-d 6 )δ10.05(s,1H),8.79(s,1H),8.02–7.99(m,1H),7.78-7.75(m,1H),7.59(d,J=7.8Hz,1H),7.55 -7.41(m,2H),6.57(d,J=7.8Hz,2H),5.69-5.63(m,1H),5.33(s,1H),4.99(d,J=10.2Hz,1H),4. 84(d,J=16.8Hz,1H),4.68(s,2H),4.26(s,1H),3.71(s,1H),3.32-3.30(m,1H),3.16(d,J=9.0H z,1H),3.01(d,J=7.8Hz,1H),2.41-2.36(m,2H),1.81(s,2H),1.46(s,6H),0.98-0.93(m,3H).D 2 O: 1 H NMR (600 MHz, DMSO-d 6)δ8.80(s,1H),8.03–7.99(m,1H),7.73(s,1H),7.60(d,J=7.2Hz,1H),7.49-7.37(m,2H), 6.59(d,J=7.2Hz,2H),5.69-5.63(m,1H),5.02(d,J=10.2Hz,1H),4.83(d,J=16.8Hz,1H), 4.68(s,2H),4.26(s,1H),3.78(s,1H),3.32(d,J=7.8Hz,1H),3.19(d,J=9.0Hz,1H),3.03 (d,J=9.0Hz,1H),2.42-2.35(m,2H),1.84(d,J=6.6Hz,2H),1.47(s,6H),1.0-0.94(m,6H).

[0545] Example 42: Synthesis of Compound 42

[0546]

[0547] Step 1: Synthesis of compound 42-1

[0548] Add 27-2 (330 mg, 1.03 mmol) and dissolve it in dichloromethane (5 ml) in a dry single-mouth bottle. Add trifluoroacetic acid (2 ml) and react at room temperature for 30 minutes. Monitor by LC-MS. After the reaction is completed, remove the solvent by distillation under reduced pressure to obtain compound 42-1 (280 mg, crude). LCMS (ESI) m / z: 220.1 [M+H] + .

[0549] Step 2: Synthesis of compound 42-2

[0550] Add substrate 42-1 (300 mg, 1.37 mmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (3 mL), add 2-iodopropane (348.92 mg, 2.05 mmol), and potassium carbonate (567.35 mg, 4.11 mmol), and react at 70°C for 12 hours. LC-MS monitoring. After the reaction is completed, the reaction solution is cooled to room temperature, water is added, and ethyl acetate is extracted three times. The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by column chromatography to obtain compound 42-2 (280 mg, 1.07 mmol). LCMS (ESI) m / z: 262.2 [M+H] + .

[0551] Step 3: Synthesis of compound 42-3

[0552] Add 42-2 (0.8 g, 3.06 mmol) to a 50 mL round-bottom flask and dissolve in methanol (10 mL). Add a catalytic amount of palladium carbon (80 mg) and stir at room temperature for 2 hours under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 42-3 (500 mg, 2.16 mmol). LCMS (ESI) m / z: 232.2 [M+H] + .

[0553] Step 4: Synthesis of compound 42

[0554] In a dry single-mouth bottle, add substrate IM-1 (42.89 mg, 120.00 μmol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (31.07 mg, 180.00 μmol) and react at room temperature for 30 minutes. Then add N,N-diisopropylethylamine (77.54 mg, 600.00 μmol) and 42-3 (33.31 mg, 144.00 μmol) to the reaction solution and react at room temperature for 4 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 42 (30 mg, 51.77 μmol). LCMS (ESI) m / z: 541.4 [M+H] + , HPLC Method BR T =7.02min,purity>93.3%. 1 H NMR (600 MHz, DMSO-d 6 )δ10.09(s,1H),8.80(s,1H),8.03(s,1H),7.76(s,1H),7.60-7.47(m,3H),6.69(d,J=8.4 Hz,2H),5.68–5.64(m,1H),5.31(s,1H),5.00–4.98(m,1H),4.86–4.79(m,1H),4.68(d,J= 5.4Hz,2H),3.71(d,J=6.0Hz,2H),3.42(d,J=11.2Hz,2H),3.24(d,J=10.8Hz,2H),2.50–2 .43(m,1H),2.37–2.35(m,1H),1.50(d,J=8.4Hz,1H),1.46(s,6H),0.88(d,J=6.0Hz,6H).

[0555] Example 43: Synthesis of Compound 43

[0556]

[0557] Step 1: Synthesis of compound 43-1

[0558] Add substrate 27-1 (0.33 g, 1.66 mmol) dissolved in methanol (4 mL) to a dry single-mouth bottle, add formaldehyde (226.77 mg, 6.66 mmol) and acetic acid (0.5 mL), and stir at room temperature for 10 minutes. Add sodium cyanoborohydride (1.41 g, 6.66 mmol) and react at room temperature for 12 hours. After the reaction is completed, dilute with water, extract three times with dichloromethane, dry the organic phase over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is purified by column chromatography to obtain compound 43-1 (300 mg, 1.41 mmol). LCMS (ESI) m / z: 213.2 [M+H] + .

[0559] Step 2: Synthesis of compound 43-2

[0560] Add 43-1 (300 mg, 1.41 mmol) and dissolve it in dichloromethane (3 ml) in a dry single-mouth bottle. Add trifluoroacetic acid (1 ml) and react at room temperature for 30 minutes. After LC-MS monitoring, the solvent was removed by vacuum distillation after the raw material disappeared to obtain compound 43-2 (0.12 g, 1.07 mmol).

[0561] Step 3: Synthesis of compound 43-3

[0562] In a dry single-mouth bottle, 43-2 (0.22 g, 1.96 mmol) was dissolved in dimethyl sulfoxide (5 ml), and p-fluoronitrobenzene (304.41 mg, 2.16 mmol) and potassium carbonate (1.36 g, 9.81 mmol) were added in sequence. The reaction mixture was stirred at 100 °C for 3 hours. LC-MS monitoring, after the reaction was completed, the mixture was diluted with water, extracted with ethyl acetate three times, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound 43-3 (260 mg, 1.11 mmol). LCMS (ESI) m / z: 234.1 [M+H] + .

[0563] Step 4: Synthesis of compound 43-4

[0564] Add 43-3 (0.3 g, 1.29 mmol) to a 50 mL round-bottom flask and dissolve in methanol (5 mL). Add a catalytic amount of palladium carbon (15 mg) and stir at room temperature for 3 hours under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 43-4 (200 mg, 983.85 μmol). LCMS (ESI) m / z: 204.2 [M+H] + .

[0565] Step 5: Synthesis of compound 43

[0566] In a dry single-mouth bottle, add substrate IM-1 (42.89 mg, 120.00 μmol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (31.07 mg, 180.00 μmol) and react at room temperature for 30 minutes. Then add N, N-diisopropylethylamine (77.54 mg, 600.00 μmol) and compound 43-4 (29.27 mg, 144.00 μmol) to the reaction solution and react at room temperature for 4 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 43 (32 mg, 55.12 μmol). LCMS (ESI) m / z: 513.2 [M+H] + ,HPLC Method BR T =6.47min,purity>88.3%. 1 H NMR (600 MHz, DMSO-d 6 )δ10.05(s,1H),8.80(s,1H),7.99–7.97(m,1H),7.74(s,1H),7.60-7.58(m,1H),7.48(s, 2H),6.45(d,J=8.4Hz,2H),5.64–5.63(m,1H),5.31(s,1H),4.99(d,J=10.2Hz,1H),4.83( d,J=17.4Hz,1H),4.68(d,J=5.6Hz,2H),4.23(d,J=5.6Hz,2H),2.97(d,J=10.8Hz,2H),2. 71(d,J=11.4Hz,2H),2.42–2.39(m,1H),2.08(s,3H),1.96(d,J=7.2Hz,1H),1.46(s,6H).

[0567] Example 44: Synthesis of Compound 44

[0568]

[0569] Step 1: Synthesis of compound 44-1

[0570] Add substrate 38-1 (300 mg, 1.37 mmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (2 mL), add 1,2-fluorobromoethane (173.72 mg, 1.37 mmol), potassium carbonate (378 mg, 2.74 mmol) and potassium iodide (45.43 mg, 273.67 μmol), and react overnight at 90°C. LC-MS monitoring. After the reaction is completed, the reaction solution is cooled to room temperature, water is added, filtered, and washed with methanol. The organic phase is concentrated under reduced pressure to obtain compound 44-1 (320 mg, 1.21 mmol). LCMS (ESI) m / z: 266.1 [M+H] + .

[0571] Step 2: Synthesis of compound 44-2

[0572] Add 44-1 (320 mg, 1.21 mmol) to a round-bottom flask and dissolve it in methanol (5 mL). Add a catalytic amount of palladium carbon (30 mg) and stir at room temperature for 2 hours under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 44-2 (200 mg, 849.98 μmol). LCMS (ESI) m / z: 236.2 [M+H] + .

[0573] Step 3: Synthesis of compound 44

[0574] In a dry single-mouth bottle, add substrate IM-1 (74 mg, 207.03 μmol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (90 mg, 414.07 μmol) and react at room temperature for 30 minutes. Then add N, N-diisopropylethylamine (133.79 mg, 1.04 mmol, 180.30 μL) and 44-2 (97.43 mg, 414.07 μmol) to the reaction solution and react at room temperature for 4 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 44 (11.5 mg, 21.12 μmol). LCMS (ESI) m / z: 545.3 [M+H] + ,HPLC method A:R T =4.95min,purity:>84.1%. 1H NMR(600MHz,DMSO-d 6) δ10.05(s,1H),8.79(s,1H),8.02–8.00(m,1H),7.74(s,1H),7.59(d,J=7.8Hz,1H),7.50(s,2H),6.57(d, J=8.4Hz,2H),5.68–5.65(m,1H),5.31(s,1H),4.99(d,J=10.2,1H),4.82(d,J=17.4Hz,1H),4.67(s,2H),4.4 9–4.42(m,1H),4.38–4.36(m,1H),4.29(s,1H),3.57(s,1H),3.35(d,J=9.0Hz,1H),3.12(d,J=9.0Hz,1H),2 .90(d,J=9.6Hz,1H),2.83–2.64(m,2H),2.53(d,J=9.6Hz,1H),1.84(d,J=9.6Hz,1H),1.79(d,J=9.0Hz,1H).

[0575] Example 45: Synthesis of Compound 45

[0576]

[0577] Step 1: Synthesis of compound 45-2

[0578] In a dry microwave tube, under nitrogen protection, add substrate 36-1 (1.13 g, 9.99 mmol), 45-1 (2.20 g, 10.99 mmol), cuprous iodide (1.90 g, 9.99 mmol), potassium carbonate (1.38 g, 9.99 mmol) and N, N'-dimethylethylenediamine (880.93 mg, 9.99 mmol), and then add 1,4-dioxane (15 mL). Under nitrogen environment, heat the reaction at 120 ° C for 3 hours. LCMS monitoring, after the reaction is completed, the reaction system is cooled to room temperature, filtered, extracted with ethyl acetate 3 times, the organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by MPLC to obtain compound 45-2 (0.8 g, 3.44 mmol). LCMS (ESI) m / z: 233.1 [M+H] + .

[0579] Step 2: Synthesis of compound 45-3

[0580] Add 45-2 (800 mg, 3.44 mmol) to a 50 mL round-bottom flask and dissolve in methanol (10 mL). Add a catalytic amount of palladium carbon (40 mg) and stir at room temperature for 3 hours under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 45-3 (500 mg, 2.47 mmol). LCMS (ESI) m / z: 203.1 [M+H] + .

[0581] Step 3: Synthesis of compound 45

[0582] In a dry single-mouth bottle, add substrate IM-1 (39 mg, 109.11 μmol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (28.25 mg, 163.67 μmol) and react at room temperature for 30 minutes. Then add N,N-diisopropylethylamine (70.51 mg, 545.56 μmol, 95.02 μL) and 45-3 (33.10 mg, 163.67 μmol) to the reaction solution and react at room temperature for 4 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 45 (32 mg, 62.55 μmol). LCMS (ESI) m / z: 512.2 [M+H] + , HPLC Method BR T =7.60min,purity>100.0%. 1 H NMR (600 MHz, DMSO-d 6 )δ10.47(s,1H),8.85(s,1H),8.41(s,1H),8.04-8.01(m,1H),7.88(d,J=7.8Hz,1H),7.79–7.68(m,2H),7.52(d,J=9.0Hz,2H),6.88(d,J =8.4Hz,2H),5.71–5.64(m,1H),5.34(s,1H),5.00(d,J=10.2Hz,1H),4.85–4.77(m,1H),4.65(d,J=6.0Hz,2H),2.97(s,6H),1.46(s,6H).

[0583] Example 46: Synthesis of Compound 46

[0584]

[0585] Step 1: Synthesis of compound 46-2

[0586] In a dry single-mouth bottle, add sodium methoxide (297 mg, 5.52 mmol), nitromethane (306.35 mg, 5.02 mmol) and methanol (8 mL), and stir the reaction mixture at room temperature for 30 minutes. Add substrate 46-1 (1 g, 5.02 mmol) and stir at room temperature overnight. After the reaction is completed, concentrate under reduced pressure and add acetic acid dropwise to adjust the pH to 7.0. Extract with dichloromethane three times. Dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is purified by column chromatography to obtain compound 46-2 (640 mg, 2.46 mmol). Step 2: Synthesis of compound 46-3

[0587] Add 46-2 (100 mg, 384.19 μmol) to a 50 mL round-bottom flask and dissolve in methanol (2 mL). Add a catalytic amount of palladium carbon (40 mg) and stir at room temperature for 2 hours under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 46-3 (80 mg, 347.4 μmol). LCMS (ESI) m / z: 236.2 [M+H] + .

[0588] Step 3: Synthesis of compound 46-4

[0589] Add substrate 46-3 (1 g, 4.34 mmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (10 mL), add potassium carbonate (1.2 g, 8.68 mmol) and chloroacetyl chloride (540.40 mg, 4.78 mmol), and react at room temperature for 5 hours. After the reaction is completed, dilute with water, extract with ethyl acetate 3 times, dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is purified by column chromatography to obtain compound 46-4 (480 mg, 1.56 mmol). LCMS (ESI) m / z: 266.1 [M+H] + .

[0590] Step 4: Synthesis of compound 46-5

[0591] Add substrate 46-4 (60 mg, 195.58 μmol) to a dry single-mouth bottle, dissolve in isopropanol (6 mL), add potassium tert-butoxide (112 mg, 998.13 μmol), and stir at room temperature for 3 hours. After the reaction is completed, concentrate under reduced pressure, extract with ethyl acetate 3 times, dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain crude product 46-5 (50 mg, crude). LCMS (ESI) m / z: 215.1 [M+H] + .

[0592] Step 5: Synthesis of compound 46-6

[0593] Add substrate 46-5 (26 mg, 96.18 μmol) to a dry single-mouth bottle, dissolve in tetrahydrofuran (4.4 mL), add 2M borane tetrahydrofuran solution (0.19 mL, 384.72 μmol), and stir at 70°C for 2 hours. After the reaction is completed, concentrate under reduced pressure, add methanol and N,N,N',N'-tetramethylethylenediamine, and stir at 78°C overnight. After the reaction is completed, concentrate under reduced pressure, extract with ethyl acetate 3 times, dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain crude product 46-6 (25 mg, crude). LCMS (ESI) m / z: 257.2 [M+H] + .

[0594] Step 6: Synthesis of compound 46-7

[0595] In a dry single-mouth bottle, 46-6 (25 mg, 97.53 μmol) was dissolved in dimethyl sulfoxide (2 ml), and p-fluoronitrobenzene (13.76 mg, 97.53 μmol) and potassium carbonate (19 mg, 137.48 μmol) were added in sequence. The reaction mixture was stirred at 100 °C for 5 hours. LC-MS monitoring, after the reaction was completed, the mixture was diluted with water, extracted with ethyl acetate three times, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound 46-7 (30 mg, 79.48 μmol). LCMS (ESI) m / z: 234.1 [M+H] + .

[0596] Step 7: Synthesis of Compound 46-8

[0597] Add 46-7 (18 mg, 47.69 μmol) dissolved in dichloromethane (1 ml) to a dry single-mouth bottle. Add trifluoroacetic acid (1 ml) and react at room temperature for 30 minutes. After the disappearance of the starting material, the solvent was removed by distillation under reduced pressure to obtain compound 46-8 (13 mg, crude).

[0598] Step 8: Synthesis of compound 46-9

[0599] Add substrate 46-8 (13 mg, 46.88 μmol) dissolved in methanol (2 mL) to a dry single-mouth bottle, add formaldehyde (16 mg, 469.76 μmol) and acetic acid (50 μL), and stir at room temperature for 10 minutes. Add sodium cyanoborohydride (10 mg, 159.13 μmol) and react at 50°C for 2 hours. After the reaction is completed, concentrate under reduced pressure, and purify the residue by MPLC to obtain compound 46-9 (10 mg, 34.32 μmol). LCMS (ESI) m / z: 262.2 [M+H] + .

[0600] Step 9: Synthesis of Compound 46-10

[0601] Add 46-9 (50 mg, 171.62 μmol) to a 50 mL round-bottom flask and dissolve in methanol (3 mL). Add a catalytic amount of palladium carbon (15 mg) and stir at room temperature for 2 hours under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 46-10 (30 mg, 114.7 μmol). LCMS (ESI) m / z: 236.2 [M+H] + .

[0602] Step 10: Synthesis of compound 46

[0603] Add substrate IM-6 (42 mg, 112.47 μmol) and tetrahydrofuran (4 mL) to a dry single-mouth bottle, stir and dissolve, then add N,N-diisopropylethylamine (72.68 mg, 562.36 μmol) and 46-10 (30 mg, 114.78 μmol), and react at room temperature for 2 hours. LC-MS monitoring. After the reaction is completed, the solvent is removed by distillation under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (basic) to obtain compound 46 (32 mg, 56.07 μmol). LCMS (ESI) m / z: 571.2 [M+H] + ,HPLC method B:R T =6.53min,purity:99.5%. 1 H NMR (600 MHz, DMSO-d 6 )δ10.15(s,1H),8.83(s,1H),8.04(s,1H),7.75(d,J=6.6Hz,1H),7.62–7.59(m,3H),6.91(d,J=8.4Hz,2H), 5.70-5.63(m,1H),5.34(s,1H),5.02–4.99(m,1H),4.83(d,J=16.8Hz,1H),4.68(d,J=5.4Hz,2H),3.81–3.74 (m,2H),3.20(d,J=12.0Hz,1H),3.16-3.14(m,1H),2.99-2.96(m,1H),2.89(d,J=12.0Hz,1H),2.66-2.64(m, 1H),2.32(s,1H),2.32-2.21(m,5H),1.74–1.66(m,2H),1.57-1.52(m,1H),1.46(s,6H),1.44–1.38(m,1H).D 2 O: 1 H NMR (600 MHz, DMSO-d 6)δ8.83(s,1H),8.05(s,1H),7.74(d,J=7.2Hz,1H),7.63–7.59(m,3H),6.92(d,J=9.0Hz,2H),5 .70-5.63(m,1H),5.02(d,J=10.2Hz,1H),4.83(d,J=17.4Hz,1H),4.69(d,J=5.4Hz,2H),3.82– 3.77(m,2H),3.18-3.13(m,2H),3.00-2.96(m,1H),2.91(d,J=12.0Hz,1H),2.67-2.64(m,1H), 2.51-2.44(m,2H),2.35-2.26(m,4H),1.71-1.67(m,2H),1.57-1.54(m,1H),1.50-1.47(m,7H).

[0604] Example 47: Synthesis of Compound 47

[0605]

[0606] Step 1: Synthesis of compound 47-1

[0607] In a dry single-mouth bottle, 4-1 (323 mg, 2 mmol) was dissolved in ethanol (10 ml), and potassium hydroxide (337 mg, 6 mmol) and p-bromobenzaldehyde (370 mg, 2 mmol) were added in sequence. The reaction mixture was stirred at room temperature overnight. After the reaction was completed, the residue was concentrated under reduced pressure and purified by MPLC to obtain compound 47-1 (456 mg, 1.6 mmol). LCMS (ESI) m / z: 292.1 [M+H] + .

[0608] Step 2: Synthesis of compound 47-2

[0609] Add trimethylsulfoxide iodide (1.6 g, 8 mmol) and sodium hydroxide (0.64 g, 16 mmol) dissolved in tetrahydrofuran (20 mL) in a single-mouth bottle, replace nitrogen, and react at room temperature for 1 hour. Add 47-1 (582 mg, 2 mmol), replace nitrogen again, stir at 65 ° C for 1 hour, and monitor the reaction by LC-MS. After the reaction is completed, concentrate under reduced pressure to remove the solvent, add ethyl acetate and water to extract 3 times, dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is purified by column chromatography to obtain compound 47-2 (284 mg, 0.93 mmol). LCMS (ESI) m / z: 306.1 [M+H] + .

[0610] Step 3: Synthesis of compound 47-3

[0611] In a dry single-mouth bottle, add substrate IM-3 (78 mg, 0.2 mol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (52 mg, 0.3 mmol) and react at room temperature for 30 minutes. Then add N, N-diisopropylethylamine (129 mg, 1 mmol) and ammonia water (0.1 mL, 0.4 mmol) to the reaction solution and react at room temperature for 2 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 47-3 (58 mg, 0.16 mmol). LCMS (ESI) m / z: 585.2 [M+H] + .

[0612] Step 4: Synthesis of compound 47

[0613] In a dry microwave tube, under nitrogen protection, add substrate 47-3 (58 mg, 0.16 mmol), 47-2 (70 mg, 0.23 mmol), cuprous iodide (61 mg, 320.29 μmol), potassium carbonate (44.2 mg, 319.83 μmol) and N, N'-dimethylethylenediamine (56 mg, 635.28 μmol), and then add 1,4-dioxane (4 mL). Under nitrogen environment, heat at 110 ° C for 6 hours. LCMS monitoring, after the reaction is completed, the reaction system is cooled to room temperature, concentrated under reduced pressure to remove the solvent, and the residue is purified by MPLC to obtain compound 47 (61 mg, 104.33 μmol). LCMS (ESI) m / z: 585.2 [M+H] + ,HPLC method B:R T =6.54min,purity:100%. 1 H NMR (600 MHz, DMSO-d 6)δ10.23(s,1H),8.86(s,1H),7.80–7.77(m,1H),7.63(d,J=8.4Hz,2H),7.30–7.25(m,3H),6.61(d,J= 8.0Hz,1H),5.65–5.62(m,1H),5.00(d,J=10.2Hz,1H),4.90(d,J=17.4Hz,1H),4.75(d,J=6.0Hz,2H), 3.37(s,6H),2.96–2.92(m,2H),2.74–2.72(m,1H),2.50–2.47(m,1H),2.44–2.43(m,1H),2.22–2.21( m,1H),1.99–1.98(m,1H),1.93–1.86(m,2H),1.83–1.82(m,1H),1.65–1.63(m,1H),1.60–1.57(m,1H).

[0614] Step 5: Synthesis of compounds 47a and 47b

[0615] Compound 47 (20 mg) was separated by SFC to give 47a (6 mg), LCMS (ESI) m / z: 585.2 [M+H] + ,HPLCmethod B:R T =6.72min, purity:98.7%. and 47b (8.6mg), LCMS (ESI) m / z: 585.2[M+H] + ,HPLCmethod B:R T =6.75min,purity:99.6%.

[0616] Example 48: Synthesis of Compound 48

[0617]

[0618] Step 1: Synthesis of compound 48-1

[0619] Add substrate 27-2 (100 mg, 456.12 μmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (2 mL), add 1-bromo-2-fluoroethane (86.86 mg, 684.18 μmol), potassium carbonate (88.25 mg, 638.57 μmol) and potassium iodide (8 mg, 48.19 μmol), and react at 100 ° C for 7.5 hours. LC-MS monitoring. After the reaction is completed, the reaction solution is cooled to room temperature and extracted with ethyl acetate three times. The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 48-1 (60 mg, 226.17 μmol). LCMS (ESI) m / z: 266.2 [M+H] + .

[0620] Step 2: Synthesis of compound 48-2

[0621] Add 48-1 (60 mg, 226.17 μmol) to a 50 mL round-bottom flask and dissolve in methanol (5 mL). Add a catalytic amount of palladium carbon (15 mg) and stir at room temperature for 1 hour under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 48-2 (45 mg, crude). LCMS (ESI) m / z: 236.2 [M+H] + .

[0622] Step 3: Synthesis of compound 48

[0623] Add substrate IM-6 (42 mg, 112.47 μmol) and tetrahydrofuran (4 mL) to a dry single-mouth bottle, stir and dissolve, then add N,N-diisopropylethylamine (72.68 mg, 562.36 μmol) and 48-2 (42 mg, 112.47 μmol), and react at room temperature for 4 hours. LC-MS monitoring. After the reaction is completed, the solvent is removed by distillation under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (basic) to obtain compound 48 (45 mg, 74.45 μmol). LCMS (ESI) m / z: 545.2 [M+H] + ,HPLC method B:R T =6.81min,purity:90.1%. 1H NMR(600MHz,Chloroform-d)δ8.81(s,1H),7.86-7.84(m,1H),7.78-7.77(m,1H),7.48(s,2H),7.33(d ,J=7.8Hz,1H),6.73(d,J=9.0Hz,2H),5.73-5.67(m,1H),5.04(d,J=10.2Hz,1H),4.94(d,J=16.8Hz,1 H),4.74(d,J=6.0Hz,2H),4.62-4.60(m,1H),4.54-4.52(m,1H),3.99(s,1H),3.92(d,J=4.8Hz,2H),3 .64(d,J=10.8Hz,2H),3.39(d,J=10.8Hz,2H),2.78-2.70(m,3H),1.68(d,J=8.4Hz,1H),1.58(s,6H).

[0624] Example 49: Synthesis of Compound 49

[0625]

[0626] Step 1: Synthesis of compound 49-1

[0627] Add substrate 27-2 (100 mg, 456.12 μmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (2 mL), add cyclobutyl bromide (92.37 mg, 684.18 μmol), potassium carbonate (158 mg, 1.14 mmol) and potassium iodide (8 mg, 48.19 μmol), and react at 100 ° C for 32 hours. LC-MS monitoring. After the reaction is completed, the reaction solution is cooled to room temperature and extracted with ethyl acetate three times. The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 49-1 (32 mg, 117.07 μmol). LCMS (ESI) m / z: 274.2 [M+H] + .

[0628] Step 2: Synthesis of compound 49-2

[0629] Add 49-1 (32 mg, 117.07 μmol) to a 50 mL round-bottom flask and dissolve in methanol (5 mL). Add a catalytic amount of palladium carbon (10 mg) and stir at room temperature for 1 hour under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 49-2 (28 mg, crude). LCMS (ESI) m / z: 244.1 [M+H] + .

[0630] Step 3: Synthesis of compound 49

[0631] Add substrate IM-6 (35.7 mg, 95.60 μmol) and tetrahydrofuran (3 mL) to a dry single-mouth bottle, stir and dissolve, then add N,N-diisopropylethylamine (61.78 mg, 478.00 μmol) and 49-2 (25.59 mg, 105.16 μmol), and react at room temperature for 4 hours. LC-MS monitoring. After the reaction is completed, the solvent is removed by distillation under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (basic) to obtain compound 49 (20.8 mg, 33.31 μmol). LCMS (ESI) m / z: 553.2 [M+H] + ,HPLC method B:R T =7.50min,purity:88.5%. 1 H NMR(600MHz,Chloroform-d)δ8.82(s,1H),7.86-7.84(m,1H),7.78-7.77(m,1H),7.48(s,2H),7.33 (d,J=7.8Hz,1H),6.71(d,J=8.4Hz,2H),5.74-5.67(m,1H),5.04(d,J=10.2Hz,1H),4.94(d,J=19.4H z,1H),4.67(d,J=5.4Hz,2H),3.94-3.88(m,3H),3.47(d,J=10.2Hz,2H),3.35(d,J=10.2Hz,2H),3. 21(s,1H),2.81-2.72(m,1H),2.00-1.90(m,5H),1.78-1.74(m,1H),1.69-1.64(m,1H),1.58(s,6H).

[0632] Example 50: Synthesis of Compound 50

[0633]

[0634] Step 1: Synthesis of compound 50-2

[0635] Add substrate 50-1 (2 g, 14.91 mmol) dissolved in methanol (40 mL) to a dry single-mouth bottle, add formaldehyde (358.09 mg, 11.92 mmol) and acetic acid (89.51 mg, 1.49 mmol), and stir at room temperature for 15 minutes. Add sodium triacetoxyborohydride (15.80 g, 74.53 mmol) and react at room temperature for 3 hours. After the reaction is completed, concentrate under reduced pressure, extract with dichloromethane three times, dry the organic phase over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is purified by MPLC to obtain compound 50-2 (200 mg, 1.35 mmol). LCMS (ESI) m / z: 149.1 [M+H] + .

[0636] Step 2: Synthesis of compound 50-3

[0637] Under nitrogen protection, 50-2 (138 mg, 0.93 mmol), p-fluoronitrobenzene (282 mg, 1.4 mmol), Pd 2 (dba) 3 (85 mg, 0.093 mmol), XantPhos (108 mg, 0.18 mmol), potassium tert-butoxide (358 mg, 3.72 mmol) and 1,4-dioxane (3 mL). The reaction mixture was stirred at 110 ° C for 3 hours under a nitrogen atmosphere. LC-MS monitoring, after the reaction was completed, it was cooled to room temperature, extracted with ethyl acetate three times, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound 50-3 (20 mg, 0.074 mmol). LCMS (ESI) m / z: 270.1 [M+H] + .

[0638] Step 3: Synthesis of compound 50-4

[0639] Add 50-3 (20 mg, 74 μmol) to a 50 mL round-bottom flask and dissolve in methanol (5 mL). Add a catalytic amount of palladium carbon (10 mg) and stir at room temperature for 1 hour under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 50-4 (7.80 mg, 32.59 μmol). LCMS (ESI) m / z: 240.2 [M+H] + .

[0640] Step 4: Synthesis of compound 50

[0641] In a dry single-mouth bottle, add substrate IM-1 (13.98 mg, 39.11 μmol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (24.75 mg, 143.41 μmol) and react at room temperature for 30 minutes. Then add N,N-diisopropylethylamine (42.12 mg, 325.93 μmol) and 50-4 (7.80 mg, 32.59 μmol) to the reaction solution and react at room temperature for 2 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 50 (2 mg, 3.65 μmol). LCMS (ESI) m / z: 549.2 [M+H] + ,HPLC method B:R T =9.17min,purity:95.7%. 1 HNMR (600 MHz, DMSO-d 6 )δ10.28(s,1H),8.87(s,1H),8.03–7.99(m,1H),7.75(d,J=8.4Hz,1H),7.70(d,J=8.4Hz,2H) ,7.61(d,J=7.8Hz,1H),7.15(d,J=8.4Hz,2H),6.70–6.60(m,2H),6.53(d,J=8.4Hz,1H),6.49 –6.42(m,1H),5.71–5.62(m,1H),5.32(s,1H),5.02–4.96(m,1H),4.85–4.79(m,1H),4.68(d, J=6.0Hz,2H),3.68–3.65(m,2H),3.30(d,J=1.8Hz,2H),2.87(s,3H),1.46(s,6H),1.24(s,1H)

[0642] Example 51: Synthesis of Compound 51

[0643]

[0644] Step 1: Synthesis of compound 51-2

[0645] In a dry microwave tube, under nitrogen protection, add substrate 51-1 (44 mg, 195.48 μmol), IM-1-4 (44 mg, 197.96 μmol), cuprous iodide (37.70 mg, 197.96 μmol), potassium carbonate (38.3 mg, 227.14 μmol) and N, N'-dimethylethylenediamine (34.90 mg, 395.92 μmol), and then add 1,4-dioxane (2 mL). Under nitrogen environment, heat the reaction at 120 ° C for 4 hours. LCMS monitoring, after the reaction is completed, the reaction system is cooled to room temperature, concentrated under reduced pressure to remove the solvent, and the residue is purified by MPLC to obtain compound 51-2 (26 mg, 70.95 μmol). LCMS (ESI) m / z: 367.1 [M+H] + .

[0646] Step 2: Synthesis of compound 51

[0647] In a dry single-mouth bottle, add substrate 51-2 (56 mg, 152.82 μmol) and tetrahydrofuran (4 mL) to dissolve. Add m-chloroperbenzoic acid (42.06 mg, 244.52 μmol) and react at 50°C for 2 hours. Then add N,N-diisopropylethylamine (98.75 mg, 764.11 μmol) and 27-4 (37.28 mg, 183.39 μmol) to the reaction solution and react at 50°C for 16 hours. Monitor by LC-MS, concentrate under reduced pressure after the reaction is completed, and purify the residue by preparative high performance liquid chromatography (basic) to obtain compound 51 (11 mg, 21.09 μmol). LCMS (ESI) m / z: 522.2 [M+H] + ,HPLC method B:R T =8.05min,purity:96.8%. 1 HNMR (600 MHz, DMSO-d 6)δ10.05(s,1H),8.80(s,1H),8.22(d,J=8.4Hz,1H),7.64–7.59(m,4H),6.67(d,J=8.4Hz, 2H),6.56(d,J=3.6Hz,1H),5.75–5.65(m,1H),5.00(d,J=10.2Hz,1H),4.86(d,J=17.4Hz,1 H),4.64(s,2H),4.30–4.26(m,2H),3.57(d,J=6.0Hz,2H),3.41(d,J=10.8Hz,2H),3.25(d ,J=11.4Hz,2H),2.42–2.41(m,1H),1.98(s,3H),1.52(d,J=8.4Hz,1H),1.40–1.38(m,3H).

[0648] Example 52: Synthesis of Compound 52

[0649]

[0650] Step 1: Synthesis of compound 52-2

[0651] Add substrate 52-1 (1 g, 3.94 mmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (10 mL), add iodomethane (1.40 g, 9.84 mmol), cesium carbonate (5.13 g, 15.74 mmol), and react at 50°C for 2 hours. Monitor by LC-MS. After the reaction, cool the reaction solution to room temperature, add water, extract with ethyl acetate three times, dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by column chromatography to obtain compound 52-2 (1 g, 3.54 mmol). LCMS (ESI) m / z: 283.1 [M+H] + .

[0652] Step 2: Synthesis of Compound 52-3

[0653] In a dry microwave tube, under nitrogen protection, add substrate 52-2 (167.55 mg, 593.88 μmol), IM-1-4 (110 mg, 494.90 μmol), cuprous iodide (9.43 mg, 49.49 μmol), potassium carbonate (95.76 mg, 692.86 μmol) and N, N'-dimethylethylenediamine (8.73 mg, 98.98 μmol), and then add N, N-dimethylformamide (2 mL). Under nitrogen environment, heat the reaction at 110 ° C for 12 hours. LCMS monitoring, after the reaction is completed, the reaction system is cooled to room temperature, concentrated under reduced pressure to remove the solvent, and the residue is purified by MPLC to obtain compound 52-3 (16 mg, 37.78 μmol). LCMS (ESI) m / z: 425.1 [M+H] + .

[0654] Step 3: Synthesis of compound 52

[0655] In a dry single-mouth bottle, add substrate 52-3 (18.00 mg, 42.50 μmol) and tetrahydrofuran (0.5 mL) to dissolve. Add m-chloroperbenzoic acid (11.00 mg, 63.76 μmol) and react at room temperature for 10 minutes. Then add N,N-diisopropylethylamine (27.47 mg, 212.52 μmol) and 27-4 (8.64 mg, 42.50 μmol) to the reaction solution and react at room temperature for 4 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (basic) to obtain compound 52 (7.2 mg, 12.44 μmol). LCMS (ESI) m / z: 579.1 [M+H] + , HPLC Method BR T =7.30min,purity>98.0%. 1 H NMR (600 MHz, DMSO-d 6)δ9.92(s,1H),8.77(s,1H),7.61(d,J=1.8Hz,1H),7.56(d,J=8.4Hz,1H),7.52(s,1H), 7.39(s,1H),7.19(s,1H),6.59(s,3H),5.68–5.66(m,1H),5.09(s,1H),4.93(s,1H),4. 24(s,2H),3.81(s,3H),3.80(s,2H),3.60(s,3H),3.56(d,J=5.4Hz,2H),3.37(d,J=10. 8Hz, 2H), 3.24–3.18 (m, 2H), 2.39 (d, J = 2.4Hz, 1H), 1.96 (s, 3H), 1.50 (d, J = 8.4Hz, 1H).

[0656] Example 53: Synthesis of Compound 53

[0657]

[0658] Step 1: Synthesis of compound 53-2

[0659] Add substrate 53-1 (2.1 g, 10.66 mmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (25 mL), add iodoethane (4.99 g, 31.97 mmol), potassium carbonate (4.22 g, 31.97 mmol), and react at 90°C for 12 hours. Monitor by LC-MS. After the reaction, cool the reaction solution to room temperature, add water, extract with ethyl acetate three times, dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by column chromatography to obtain compound 53-2 (1.38 g, 6.13 mmol). LCMS (ESI) m / z: 226.0 [M+H] + .

[0660] Step 2: Synthesis of compound 53-3

[0661] Under nitrogen protection, 53-2 (849 mg, 8.66 mmol), B (pin) were added into a dry single-mouth bottle. 2 (1.61 g, 6.35 mmol), Pd(dppf)Cl 2(236 mg, 0.29 mmol), potassium tert-butoxide (845 mg, 8.66 mmol) and 1,4-dioxane (30 mL). The reaction mixture was stirred at 110 ° C for 16 hours under a nitrogen atmosphere. LC-MS monitoring, after the reaction was completed, it was cooled to room temperature, extracted with ethyl acetate three times, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound 53-3 (680 mg, 2.5 mmol). LCMS (ESI) m / z: 274.2 [M+H] + .

[0662] Step 3: Synthesis of compound 53-4

[0663] Add substrate 53-3 (0.47 g, 1.73 mmol) to a dry single-mouth bottle and dissolve in ethanol (12 mL). Add 6M hydrochloric acid solution (0.3 mL, 1.73 mmol) and react at 90°C for 7 hours. Monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure and purify the residue by MPLC to obtain compound 53-4 (180 mg, 0.95 mmol). LCMS (ESI) m / z: 192.1 [M+H] + .

[0664] Step 4: Synthesis of compound 53-5

[0665] Add substrate 53-4 (217.99 mg, 1.15 mmol) to a dry single-mouth bottle and dissolve in dichloromethane (20 mL). Add IM-1-4 (170 mg, 764.85 μmol), pyridine (242.00 mg, 3.06 mmol), copper acetate (277.84 mg, 1.53 mmol) and stir at room temperature for 24 hours. Monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure and purify the residue by MPLC to obtain compound 53-5 (31 mg, 84.60 μmol). LCMS (ESI) m / z: 368.1 [M+H] + .

[0666] Step 5: Synthesis of compound 53

[0667] In a dry single-mouth bottle, add substrate 53-5 (39 mg, 106.43 μmol) and tetrahydrofuran (6 mL) to dissolve. Add m-chloroperbenzoic acid (20.20 mg, 117.07 μmol) and react at room temperature for 30 minutes. Then add N,N-diisopropylethylamine (137.55 mg, 1.06 mmol) and 27-4 (23.80 mg, 117.07 μmol) to the reaction solution and react at room temperature for 2 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (basic) to obtain compound 53 (8.3 mg, 15.91 μmol). LCMS (ESI) m / z: 523.3 [M+H] + ,HPLC method B:R T =7.18min,purity:98.3%. 1 HNMR (600 MHz, DMSO-d 6 )δ10.14(s,1H),8.80(s,1H),8.14(s,1H),7.92(d,J=8.4Hz,1H),7.80(s,1H),7.56–7.43(m ,2H),7.28(s,1H),6.60(s,2H),5.76–5.62(m,1H),5.08(d,J=10.2Hz,1H),4.95(d,J=17.4H z,1H),4.51–4.47(m,2H),4.35(s,1H),3.56(d,J=5.4Hz,2H),3.38(d,J=10.8Hz,2H),3.22( d,J=11.2Hz,2H),2.45–2.38(m,1H),1.96(s,3H),1.50(d,J=8.4Hz,1H),1.44–1.37(m,3H).

[0668] Example 54: Synthesis of Compound 54

[0669]

[0670] Step 1: Synthesis of compound 54-2

[0671] Add substrate 54-1 (300 mg, 1.60 mmol) to a dry single-mouth bottle, dissolve in tetrahydrofuran (3 mL), add N,N'-carbonyldiimidazole (344.52 mg, 2.39 mmol), and react at 80°C for 2 hours. Monitor by LC-MS. After the reaction, cool the reaction solution to room temperature, add water, extract with ethyl acetate three times, dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is purified by column chromatography to obtain compound 54-2 (260 mg, 1.22 mmol). LCMS (ESI) m / z: 213.9 [M+H] + .

[0672] Step 2: Synthesis of compound 54-3

[0673] Add substrate 54-2 (620 mg, 2.90 mmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (6 mL), add iodoethane (451.83 mg, 2.90 mmol), cesium carbonate (1.12 g, 3.19 mmol), and react at room temperature for 12 hours. Monitor by LC-MS. After the reaction, cool the reaction solution to room temperature, add water, extract with ethyl acetate three times, dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by MPLC to obtain compound 54-3 (279 mg, 1.15 mmol). LCMS (ESI) m / z: 242.0 [M+H] + .

[0674] Step 3: Synthesis of compound 54-4

[0675] Under nitrogen protection, 54-3 (220 mg, 908.83 μmol), B (pin) were added into a dry single-mouth bottle. 2 (346.18mg,1.36mmol), Pd(dppf)Cl 2 ·Dichloromethane (74.22 mg, 90.88 μmol), potassium carbonate (408.20 mg, 2.95 mmol) and 1,4-dioxane (7 mL). The reaction mixture was stirred at 90 ° C for 16 hours under a nitrogen atmosphere. LC-MS monitoring, after the reaction was completed, it was cooled to room temperature, water was added, and ethyl acetate was extracted three times. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound 54-4 (140 mg, 484.20 μmol). LCMS (ESI) m / z: 290.2 [M+H] + .

[0676] Step 4: Synthesis of compound 54-5

[0677] Substrate 54-4 (140 mg, 484.20 μmol) was added to a dry single-mouth bottle and dissolved in acetonitrile (2 mL). 6M hydrochloric acid solution (80.70 μL) was added and reacted at 80°C for 4 hours. LC-MS monitoring. After the reaction was completed, the mixture was concentrated under reduced pressure and the residue was purified by MPLC to obtain compound 54-5 (33 mg, 159.43 μmol). LCMS (ESI) m / z: 208.1 [M+H] + .

[0678] Step 5: Synthesis of compound 54-6

[0679] Add substrate 54-5 (30 mg, 144.93 μmol) to a dry single-mouth bottle and dissolve it in dichloromethane (3 mL). Add IM-1-4 (32.2 mg, 222.27 μmol), pyridine (11.46 mg, 144.93 μmol), copper acetate (52.65 mg, 289.87 μmol) and stir at room temperature for 24 hours. Monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure and purify the residue by MPLC to obtain compound 54-6 (30 mg, 78.24 μmol). LCMS (ESI) m / z: 384.1 [M+H] + .

[0680] Step 6: Synthesis of compound 54

[0681] In a dry single-mouth bottle, add substrate 54-6 (30 mg, 78.24 μmol) and dissolve in tetrahydrofuran (2 mL). Add m-chloroperbenzoic acid (20.25 mg, 117.36 μmol) and react at room temperature for 30 minutes. Then add N,N-diisopropylethylamine (50.56 mg, 391.21 μmol) and 27-4 (19.09 mg, 93.89 μmol) to the reaction solution and react at room temperature for 1.5 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 54 (4.3 mg, 6.99 μmol). LCMS (ESI) m / z: 539.1 [M+H] + ,HPLC method A:R T =7.01min,purity:89.4%. 1 H NMR (600 MHz, DMSO-d 6)δ10.01(s,1H),8.79(s,1H),7.53-7.48(m,4H),7.26–7.24(m,1H),6.63-6.62(m, 2H),5.71-5.66(m,1H),5.08(d,J=10.2Hz,1H),4.98-4.95(m,1H),4.28-4.19(m,2H ),3.89-3.86(m,2H),3.56(d,J=4.8Hz,2H),3.38(d,J=10.8Hz,2H),3.22(d,J=10. 8Hz,2H),2.41-2.38(m,1H),1.96(s,3H),1.50(d,J=8.4Hz,1H),1.27–1.24(m,3H).

[0682] Example 55: Synthesis of Compound 55

[0683]

[0684] Step 1: Synthesis of compound 55-2

[0685] In a dry single-mouth bottle, add substrate 55-1 (0.83 g, 4.28 mmol), add N,N-dimethylformamide (6 mL) to dissolve. Add potassium tert-butoxide (960.29 mg, 8.56 mmol) and triphenylmethane chloride (1.43 g, 5.13 mmol), and react at room temperature overnight. LC-MS monitoring, after the reaction is completed, cool to room temperature, add water, extract three times with dichloromethane, dry the organic phase over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is recrystallized to obtain compound 55-2 (1.53 g, 3.51 mmol). LCMS (ESI) m / z: 437.1 [M+H] + .

[0686] Step 2: Synthesis of compound 55-3

[0687] In a dry microwave tube, under nitrogen protection, add substrate 55-2 (0.88 g, 2.02 mmol), tetrahydropyrrole (573.81 mg, 8.07 mmol), cuprous iodide (38.41 mg, 201.70 μmol), potassium tert-butoxide (452.67 mg, 4.03 mmol) and 2-acetylcyclohexanone (56.55 mg, 403.40 μmol), and then add N,N-dimethylformamide (5 mL). Under nitrogen environment, heat the reaction at 80 ° C for 24 hours. LCMS monitoring, after the reaction is completed, the reaction system is cooled to room temperature, concentrated under reduced pressure to remove the solvent, and the residue is purified by MPLC to obtain compound 55-3 (250 mg, 658.77 μmol). LCMS (ESI) m / z: 380.2 [M+H]+ .

[0688] Step 3: Synthesis of compound 55-4

[0689] Add 55-3 (140 mg, 368.91 μmol) to a dry single-mouth bottle and dissolve it in dichloromethane (2 ml). Add trifluoroacetic acid (1 ml) and react at room temperature for 30 minutes. After the disappearance of the starting material, the solvent was removed by distillation under reduced pressure to obtain compound 55-4 (22 mg, 160.37 μmol). LCMS (ESI) m / z: 138.1 [M+H] + .

[0690] Step 4: Synthesis of compound 55-5

[0691] In a dry single-mouth bottle, 55-4 (250 mg, 1.82 mmol) was dissolved in dimethyl sulfoxide (5 ml), and p-fluoronitrobenzene (282.85 mg, 2.00 mmol) and potassium carbonate (2.52 g, 18.22 mmol) were added in sequence. The reaction mixture was stirred at 100 °C for 12 hours. LC-MS monitoring, after the reaction was completed, it was diluted with water, extracted with ethyl acetate three times, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound 55-5 (200 mg, 774.37 μmol). LCMS (ESI) m / z: 259.1 [M+H] + .

[0692] Step 5: Synthesis of compound 55-6

[0693] In a dry single-mouth bottle, 55-5 (9 mg, 34.85 μmol) was dissolved in tetrahydrofuran (0.5) and acetic acid (0.1 mL), and zinc powder (45.57 mg, 696.93 μmol) was added and stirred at room temperature for 30 minutes. LC-MS monitoring, after the reaction was completed, the filtrate was filtered with diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain compound 55-6 (6 mg, 26.28 μmol). LCMS (ESI) m / z: 229.2 [M+H] + .

[0694] Step 6: Synthesis of compound 55

[0695] In a dry single-mouth bottle, add substrate IM-1 (42.27 mg, 118.27 μmol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (30.61 mg, 177.40 μmol) and react at room temperature for 30 minutes. Then add N,N-diisopropylethylamine (76.43 mg, 591.35 μmol) and 55-6 (27 mg, 118.27 μmol) to the reaction solution and react at room temperature for 4 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 55 (2 mg, 3.72 μmol). LCMS (ESI) m / z: 538.2 [M+H] + , HPLC Method BR T =8.23min,purity>97.9%. 1 HNMR (600 MHz, DMSO-d 6 )δ10.39(s,1H),8.90(s,1H),8.10(s,1H),7.82(d,J=8.4Hz,2H),7.78-7 .77(m,2H),7.74–7.69(m,2H),7.64(d,J=7.8Hz,1H),7.35(s,1H),5.72– 5.64(m,1H),5.34(s,1H),5.00(d,J=10.2Hz,1H),4.83(d,J=17.4Hz,1H) ,4.69(d,J=6.0Hz,2H),3.08–3.06(s,4H),1.92–1.89(m,4H),1.47(s,6H)

[0696] Example 56: Synthesis of Compound 56

[0697]

[0698] Step 1: Synthesis of compound 56

[0699] In a dry single-mouth bottle, add substrate 52 (2 mg, 3.46 μmol) and methanol (1 mL) to dissolve. Add 1M NaOH solution (5 μl, 5.18 μmol) and stir at room temperature for 30 minutes. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 56 (1.2 mg, 2.13 μmol). LCMS (ESI) m / z: 565.3 [M+H] + , HPLC Rt=4.68min, purity>83.3%. 1 H NMR (600 MHz, Methanol-d 4)δ8.65(s,1H),7.60(d,J=2.1Hz,1H),7.37(d,J=8.6Hz,3H),7.10(s,1H),7.07(d,J= 8.2Hz,1H),6.52(d,J=14.6Hz,2H),5.67–5.59(m,1H),5.06–5.00(m,1H),4.87(d,J= 17.4Hz,1H),4.26(d,J=5.5Hz,2H),3.73(s,3H),3.57(s,2H),3.50(d,J=2.8Hz,2H), 3.39(d,J=11.0Hz,2H),2.46(s,1H),1.84(s,2H),1.79(s,3H),1.27(d,J=2.9Hz,1H).

[0700] Example 57: Synthesis of Compound 57

[0701]

[0702] Step 1: Synthesis of Compound 57-2

[0703] In a dry microwave tube, under nitrogen protection, add substrate 57-1 (546.59 mg, 2.39 mmol), IM-1-4 (444 mg, 2.00 mmol), cuprous iodide (762 mg, 4.00 mmol), potassium carbonate (552 mg, 2.00 mmol) and N, N'-dimethylethylenediamine (705 mg, 2.00 mmol), and then add 1,4-dioxane (10 mL). Under nitrogen environment, heat the reaction at 110 ° C for 16 hours. LCMS monitoring, after the reaction is completed, the reaction system is cooled to room temperature, filtered, and the filtrate is concentrated under reduced pressure. The residue is purified by MPLC to obtain compound 57-2 (140 mg, 377.94 μmol). LCMS (ESI) m / z: 371.1 [M+H] + .

[0704] Step 2: Synthesis of compound 57

[0705] In a dry single-mouth bottle, add substrate 57-2 (7.4 mg, 19.98 μmol) and tetrahydrofuran (0.5 mL) to dissolve. Add m-chloroperbenzoic acid (7.64 mg, 44.27 μmol) and react at room temperature for 10 minutes. Then add N,N-diisopropylethylamine (5.72 mg, 44.27 μmol) and 27-4 (6 mg, 29.52 μmol) to the reaction solution and react at room temperature for 4 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 57 (8 mg, 15.22 μmol). LCMS (ESI) m / z: 526.3 [M+H] + ,HPLC R T =6.38min,purity>94.2%. 1 H NMR (600 MHz, DMSO-d 6 )δ9.95(s,1H),8.76(s,1H),7.54(s,2H),7.38(d,J=9.0Hz,1H),7.27(d,J=8.4Hz,1H),6.99(d,J=8.5Hz,1H ),6.64(d,J=8.6Hz,2H),5.69–5.67(m,1H),5.62(d,J=18.0Hz,1H),5.10(d,J=10.2Hz,1H),4.94(d,J=17.4H z,1H),4.55(s,1H),4.41(d,J=9.6Hz,1H),4.36(d,J=9.6Hz,1H),4.22(s,1H),3.55(d,J=6.0Hz,2H),3.39(d ,J=11.2Hz,2H),3.22(d,J=11.2Hz,2H),2.43–2.38(m,1H),1.96(s,3H),1.57(s,3H),1.50(d,J=8.4Hz,1H).

[0706] Example 58: Synthesis of Compound 58

[0707]

[0708] Step 1: Synthesis of compound 58-1

[0709] Add substrate 38-1 (65.29 mg, 297.82 μmol) dissolved in 1,2-dichloroethane (5 mL) to a dry single-mouth bottle, add 3-oxetanone (108 mg, 1.5 mmol) and acetic acid (0.5 mL), and stir at room temperature for 15 minutes. Add sodium triacetoxyborohydride (315.60 mg, 1.49 mmol) and react at room temperature for 10 hours. After the reaction is completed, concentrate under reduced pressure, and purify the residue by MPLC to obtain compound 58-1 (60 mg, 217.94 μmol). LCMS (ESI) m / z: 276.1 [M+H] + .

[0710] Step 2: Synthesis of compound 58-2

[0711] Add 58-1 (60 mg, 217.94 μmol) to a 50 mL round-bottom flask and dissolve in methanol (5 mL). Add a catalytic amount of palladium carbon (15 mg) and stir at room temperature for 1 hour under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 58-2 (45 mg, 183.43 μmol). LCMS (ESI) m / z: 246.2 [M+H] + .

[0712] Step 3: Synthesis of compound 58

[0713] Add substrate IM-6 (41.84 mg, 112.04 μmol) and tetrahydrofuran (2 mL) to a dry single-mouth bottle, stir and dissolve, then add N,N-diisopropylethylamine (72.40 mg, 560.20 μmol) and 58-2 (30.23 mg, 123.24 μmol), and react at room temperature for 2 hours. LC-MS monitoring. After the reaction is completed, the solvent is removed by distillation under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (basic) to obtain compound 58 (35 mg, 62.16 μmol). LCMS (ESI) m / z: 555.2 [M+H] + ,HPLC method B:R T =6.27min,purity:98.5%. 1 H NMR (600 MHz, DMSO-d 6)δ10.10(s,1H),8.80(s,1H),8.03(s,1H),7.77(s,1H),7.61-7.47(m,3H),6.67(d,J=7. 8Hz,2H),5.70-5.63(m,1H),5.32(s,1H),4.99(d,J=10.2Hz,1H),4.83(d,J=16.8Hz,1H) ,4.68(d,J=2.4Hz,2H),4.58–4.56(m,2H),4.28–4.26m,2H),3.75(d,J=6.0Hz,2H),3.62 -3.60(m,1H),3.28-3.26(m,4H),2.48-2.47(m,1H),1.54(d,J=8.4Hz,1H),1.47(s,6H).D 2 O: 1 H NMR (600 MHz, DMSO-d 6 )δ8.81(s,1H),8.04(s,1H),7.75(s,1H),7.62-7.45(s,3H),6.69(d,J=8.4Hz,2H ),5.70-5.63(m,1H),5.03(d,J=10.2Hz,1H),4.83(d,J=16.8Hz,1H),4.69(d,J=3. 0Hz,2H),4.60–4.58(m,2H),4.32–4.29(m,2H),3.76(d,J=5.4Hz,2H),3.63-3.62 (m,1H),3.26–3.24(m,4H),2.52-2.51(m,1H),1.53(d,J=8.4Hz,1H),1.48(s,6H).

[0714] Example 59: Synthesis of Compound 59

[0715]

[0716] Step 1: Synthesis of compound 59-1

[0717] Add substrate 46-6 (250 mg, 975.27 μmol) dissolved in methanol (2 mL) to a dry single-mouth bottle, add formaldehyde (16 mg, 469.76 μmol) and acetic acid (0.5 mL), and stir at room temperature for 30 minutes. Add sodium cyanoborohydride (306.43 mg, 4.88 mmol) and react at 50 ° C for 2 hours. Monitor by TLC. After the reaction is completed, concentrate under reduced pressure, and the residue is purified by column chromatography to obtain compound 59-1 (130 mg, 480.83 μmol). LCMS (ESI) m / z: 262.2 [M+H] + .

[0718] Step 2: Synthesis of compound 59-2

[0719] Add 59-1 (54 mg, 199.73 μmol) to a dry single-mouth bottle and dissolve it in dichloromethane (2 ml). Add trifluoroacetic acid (1 ml) and react at room temperature for 30 minutes. After LC-MS monitoring, the solvent is removed by vacuum distillation after the raw material disappears to obtain compound 59-2 (30 mg, 176.21 μmol).

[0720] Step 3: Synthesis of compound 59-3

[0721] In a dry single-mouth bottle, 59-2 (30 mg, 176.21 μmol) was dissolved in dimethyl sulfoxide (2 ml), and p-fluoronitrobenzene (24.86 mg, 176.21 μmol) and NaH (21.15 mg, 881.05 μmol) were added in sequence. The reaction mixture was stirred at 100 °C for 3 hours. LC-MS monitoring, after the reaction was completed, the reaction was quenched with water, extracted with ethyl acetate three times, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound 59-3 (35 mg, 120.13 μmol). LCMS (ESI) m / z: 234.1 [M+H] + .

[0722] Step 4: Synthesis of compound 59-4

[0723] Add 59-3 (35 mg, 120.13 μmol) to a 50 mL round-bottom flask and dissolve in methanol (5 mL). Add a catalytic amount of palladium carbon (15 mg) and stir at room temperature for 2 hours under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 59-4 (30 mg, 114.78 μmol). LCMS (ESI) m / z: 262.2 [M+H] + .

[0724] Step 5: Synthesis of compound 59

[0725] Add substrate IM-6 (42.86 mg, 114.78 μmol) and tetrahydrofuran (2 mL) to a dry single-mouth bottle, stir and dissolve, then add N,N-diisopropylethylamine (74.17 mg, 573.92 μmol) and 59-4 (30 mg, 114.78 μmol), and react at room temperature for 2 hours. LC-MS monitoring. After the reaction is completed, the solvent is removed by distillation under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (basic) to obtain compound 59 (3 mg, 4.75 μmol). LCMS (ESI) m / z: 571.2 [M+H] + ,HPLC method B:RT =7.61min,purity:90.3%. 1 H NMR(600MHz,Chloroform-d)δ8.82(s,1H),7.89-7.76(m,2H),7.45-7.44(m,2H) ,7.35(d,J=7.8Hz,1H),6.97(s,2H),5.73-5.67(m,1H),5.04(d,J=10.2Hz,1H), 4.94(d,J=16.8,1.2Hz,1H),4.74(d,J=6.0Hz,2H),3.97-3.83(m,3H),3.25-3.1 1(m,3H),2.58-2.33(m,6H),1.93(s,1H),1.78(s,1H),1.61(s,3H),1.58(s,6H).

[0726] Example 60: Synthesis of Compound 60

[0727]

[0728] Step 1: Synthesis of compound 60-1

[0729] IM-1-5 (250 mg, 1.16 mmol) was dissolved in tetrahydrofuran (2 ml) in a dry single-mouth bottle, the temperature was lowered to 0°C, NaH (83.30 mg, 3.47 mmol) was added, and the mixture was stirred at 0°C for 10 minutes. Iodomethane (492.7 mg, 3.47 mmol) was added, the mixture was warmed to room temperature and stirred for 6 hours. LC-MS monitoring was performed, and the reaction was quenched with water after completion, extracted with ethyl acetate three times, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound 60-1 (210 mg, 912.64 μmol). LCMS (ESI) m / z: 230.1 [M+H] + .

[0730] Step 2: Synthesis of compound 60-2

[0731] In a dry microwave tube, under nitrogen protection, add substrate 60-1 (207.05 mg, 899.82 μmol), IM-1-4 (200 mg, 899.82 μmol), cuprous iodide (342.74 mg, 1.80 mmol), potassium carbonate (174.10 mg, 1.26 mmol) and N, N'-dimethylethylenediamine (705 mg, 2.00 mmol), and then add 1,4-dioxane (4 mL). Under nitrogen environment, heat at 110 ° C for 2 hours. LC-MS monitoring, after the reaction is completed, the reaction system is cooled to room temperature, ammonia water (2 mL) is added, and ethyl acetate is extracted three times. The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by MPLC to obtain compound 60-2 (80 mg, 215.37 μmol). LCMS (ESI) m / z: 372.2 [M+H] + .

[0732] Step 3: Synthesis of compound 60

[0733] In a dry single-mouth bottle, add substrate 60-2 (46 mg, 123.84 μmol) and tetrahydrofuran (0.5 mL) to dissolve. Add m-chloroperbenzoic acid (32 mg, 185.76 μmol) and react at room temperature for 10 minutes. Then add N,N-diisopropylethylamine (80.02 mg, 619.19 μmol) and 27-4 (30.21 mg, 148.60 μmol) to the reaction solution and react at room temperature for 6 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (basic) to obtain compound 60 (14.08 mg, 26.74 μmol). LCMS (ESI) m / z: 527.2 [M+H] + ,HPLC method B:R T =5.86min,purity:97.6%. 1 H NMR (600 MHz, DMSO-d 6)δ10.10(s,1H),8.80(s,1H),8.07-8.05(m,1H),7.83-7.82(m,1H),7.59(s,2H),7.46(d ,J=7.8Hz,1H),6.72(d,J=7.8Hz,2H),5.70-5.63(m,1H),4.99(d,J=10.2Hz,1H),4.84-4. 81(m,1H),4.69(d,J=4.2Hz,2H),3.59(d,J=3.6Hz,2H),3.43(d,J=10.8Hz,2H),3.28(d,J =10.8Hz,2H),3.09(s,3H),2.43(s,1H),1.99(s,3H),1.54(d,J=7.8Hz,1H),1.49(s,6H).

[0734] Example 61: Synthesis of Compound 61

[0735]

[0736] Step 1: Synthesis of compound 61-2

[0737] Add substrate 61-1 (200 mg, 1.30 mmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (2.5 mL), add iodoethane (406.24 mg, 2.60 mmol), potassium carbonate (5.4 g, 3.91 mmol), and react at 75°C for 1 hour. Monitor by LC-MS. After the reaction, cool the reaction solution to room temperature, filter, and concentrate under reduced pressure. The residue is purified by MPLC to obtain compound 61-2 (140 mg, 770.83 μmol). LCMS (ESI) m / z: 266.2 [M+H] + .

[0738] Step 2: Synthesis of compound 61-3

[0739] Substrate 61-2 (130 mg, 715.77 μmol) was added to a dry single-mouth bottle, dissolved in acetonitrile (2 mL), and trimethylsilyl bromide (109.58 mg, 715.77 μmol) was added. The reaction was allowed to react at 90°C for 4 hours. LC-MS monitoring was performed. After the reaction was completed, the reaction solution was cooled to room temperature, concentrated under reduced pressure, and the residue was purified by MPLC to obtain compound 61-3 (60 mg, 265.40 μmol). LCMS (ESI) m / z: 266.2 [M+H] + .

[0740] Step 3: Synthesis of compound 61-4

[0741] In a dry microwave tube, under nitrogen protection, add substrate 61-3 (36.99 mg, 163.60 μmol), IM-1-4 (40 mg, 179.96 μmol), cuprous iodide (62.32 mg, 327.21 μmol), potassium carbonate (31.65 mg, 229.05 μmol) and N, N'-dimethylethylenediamine (57.69 mg, 654.42 μmol), and then add 1,4-dioxane (4 mL). Under nitrogen environment, heat at 110 ° C for 3 hours. LCMS monitoring, after the reaction is completed, the reaction system is cooled to room temperature, concentrated under reduced pressure, and the residue is purified by MPLC to obtain compound 61-4 (19 mg, 51.71 μmol).

[0742] Step 4: Synthesis of compound 61

[0743] In a dry single-mouth bottle, add substrate 61-4 (19 mg, 51.71 μmol) and tetrahydrofuran (0.7 mL) to dissolve. Add m-chloroperbenzoic acid (13.39 mg, 77.57 μmol) and react at room temperature for 30 minutes. Then add N,N-diisopropylethylamine (33.42 mg, 258.55 μmol) and 27-4 (10.51 mg, 51.71 μmol) to the reaction solution and react at room temperature for 10 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (basic) to obtain compound 61 (3.89 mg, 7.44 μmol). LCMS (ESI) m / z: 523.3 [M+H] + ,HPLC method A:R T =4.98min,purity:98.6%. 1 H NMR (600MHz, CDCl3) δ8.76 (s, 1H), 8.05 (d, J = 8.4Hz, 1H), 7.89 (s, 1H), 7.78 (d, J = 8. 4Hz,1H),7.44(s,2H),6.67(d,J=8.4Hz,2H),5.63-5.59(m,1H),4.93-4.90(m,3H),4 .87–4.84(m,1H),4.46–4.42(m,2H),3.95(s,2H),3.58(d,J=8.4Hz,2H),3.46(d,J=8 .4Hz,2H),2.93-2.87(m,1H),2.25(s,3H),1.71(d,J=6.0Hz,1H),1.61–1.59(m,3H).

[0744] Example 62: Synthesis of Compound 62

[0745]

[0746] Step 1: Synthesis of compound 62

[0747] In a dry microwave tube, under nitrogen protection, substrate 62-1 (36.99 mg, 163.60 μmol), 62-2 (40.39 mg, 123.75 μmol), cuprous iodide (47.14 mg, 247.50 μmol), potassium carbonate (68.41 mg, 495.01 μmol) and N,N'-dimethylethylenediamine (21.78 mg, 247.50 μmol) were added, and then 1,4-dioxane (2 mL) was added. Under nitrogen environment, the reaction was heated at 110 ° C for 16 hours. LC-MS monitoring, after the reaction was completed, the reaction system was cooled to room temperature, concentrated under reduced pressure, and the residue was purified by MPLC to obtain compound 62 (0.7 mg, 1.23 μmol). LCMS (ESI) m / z: 569.3 [M+H] + ,HPLC methodA:R T =6.03min,purity:73.2%. 1 H NMR (600MHz, DMSO-d6) δ8.93–8.92(m,1H),8.07–8.03(m,1H),7.92–7.87(m,4H),7.88(d,J=7. 8Hz,1H),7.72(d,J=7.8Hz,1H),5.76–5.71(m,1H),5.03–5.01(m,1H),4.92–4.89(m,1H),4.83 –4.82(m,2H),4.66–4.63(m,1H),4.43–4.41(m,1H),4.29–4.26(m,1H),3.57(d,J=11.4Hz,2H) ,3.04–3.00(m,2H),2.95(s,3H),2.19(d,J=14.4Hz,1H),2.16–1.51(m,4H),1.58–1.53(m,7H)

[0748] Example 63: Synthesis of Compound 63

[0749]

[0750] Step 1: Synthesis of compound 63-2

[0751] Add substrate 38-1 (44 mg, 200.69 μmol) dissolved in 1,2-dichloroethane (5 mL) to a dry single-mouth bottle, add 63-1 (171.79 mg, 1.00 mmol) and acetic acid (0.5 mL), and stir at room temperature for 15 minutes. Add sodium triacetoxyborohydride (211.94 mg, 1 mmol) and react at room temperature for 12 hours. After the reaction is completed, concentrate under reduced pressure, and purify the residue by MPLC to obtain compound 63-2 ((350 mg, 934.75 μmol). LCMS (ESI) m / z: 375.2 [M+H] + .

[0752] Step 2: Synthesis of compound 63-3

[0753] Add 63-2 (60 mg, 160.24 μmol) to a dry single-mouth bottle and dissolve it in dichloromethane (2 ml). Add trifluoroacetic acid (1 ml) and react at room temperature for 30 minutes. After the disappearance of the starting material, the solvent was removed by distillation under reduced pressure to obtain compound 63-3 (43 mg, 156.75 μmol). LCMS (ESI) m / z: 275.2 [M+H] + .

[0754] Step 3: Synthesis of compound 63-4

[0755] Add substrate 63-3 (43 mg, 156.75 μmol) dissolved in methanol (3 mL) to a dry single-mouth bottle, add formaldehyde (26.69 mg, 783.76 μmol) and acetic acid (0.2 mL), and stir at room temperature for 10 minutes. Add sodium cyanoborohydride (19.70 mg, 313.51 μmol) and react at 50 ° C for 6 hours. LC-MS monitoring. After the reaction is completed, concentrate under reduced pressure, extract with ethyl acetate, dry the organic phase over anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain compound 63-4 (35 mg, 121.38 μmol). LCMS (ESI) m / z: 289.2 [M+H] + .

[0756] Step 4: Synthesis of compound 63-5

[0757] Add 63-4 (35 mg, 120.13 μmol) to a 50 mL round-bottom flask and dissolve in methanol (3 mL). Add a catalytic amount of palladium carbon (15 mg) and stir at room temperature for 1 hour under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 63-5 (28 mg, 71.90 μmol). LCMS (ESI) m / z: 259.2 [M+H] + .

[0758] Step 5: Synthesis of compound 63

[0759] Add substrate IM-6 (16.89 mg, 65.36 μmol) and tetrahydrofuran (2 mL) to a dry single-mouth bottle, stir and dissolve, then add N,N-diisopropylethylamine (42.24 mg, 326.82 μmol) and 63-5 (28 mg, 71.90 μmol), and react at room temperature for 4 hours. LC-MS monitoring. After the reaction is completed, the solvent is removed by distillation under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (basic) to obtain compound 63 (11 mg, 18.49 μmol). LCMS (ESI) m / z: 568.4 [M+H] + ,HPLC method B:R T =5.21min,purity:97.5%. 1 H NMR (600 MHz, DMSO-d 6 )δ8.82-8.79(m,1H),7.99-7.91(m,1H),7.79-7.71(m,1H),7.59-7.55(m,1H),7.5 1-7.47(m,2H),6.71-6.69(m,2H),5.71-5.69(m,1H),5.67-5.61(m,1H),5.00-4.9 8(m,1H),4.83-4.56(m,3H),4.25–4.13(m,2H),4.02–3.89(m,4H),3.75–3.57(m,5 H),3.28-3.21(m,3H),,2.42-2.38(m,1H),1.80-1.72(m,1H),1.46-1.42(m,6H).D 2 O: 1 H NMR (600 MHz, 333 K, DMSO-d 6 )δ8.77(s,1H),7.94(s,1H),7.74(s,1H),7.57(d,J=7.2Hz,1H),7.47(s,2H),6.72(d,J=9.0Hz,2H),5.68-5.62(m,1H),5.00(d,J=10.2H z,1H),4.85(d,J=17.4Hz,1H),4.67(s,2H),4.17–3.62(m,10H),3.39–3.37(m,1H),3.18(s,3H),2.45(s,1H),1.77(s,1H),1.47(s,6H).

[0760] Example 64: Synthesis of Compound 64

[0761]

[0762] Step 1: Synthesis of compound 64-1

[0763] Substrate 38-1 (109 mg, 497.17 μmol) was added to a dry single-mouth bottle, dissolved in acetone (3 mL), allyl bromide (90.1 mg, 745.76 μmol) and potassium carbonate (207 mg, 745.76 μmol) were added, and reacted at room temperature for 2 hours under nitrogen atmosphere. LC-MS monitoring. After the reaction was completed, it was concentrated under reduced pressure, and the residue was purified by MPLC to obtain compound 64-1 (128 mg, 497.03 μmol). LCMS (ESI) m / z: 260.2 [M+H] + .

[0764] Step 2: Synthesis of compound 64-2

[0765] Add 64-1 (128 mg, 497.03 μmol) to a dry single-mouth bottle and dissolve it in ethanol (10 mL). Add iron powder (138.92 mg, 2.49 mmol) and ammonium chloride (399 mg, 7.46 mmol), and heat the reaction to 80 ° C and stir for 2 hours. LC-MS monitoring. After the reaction is completed, cool to room temperature, filter with diatomaceous earth, and concentrate the filtrate under reduced pressure. The residue is purified by MPLC to obtain compound 64-2 (25 mg, 109.02 μmol). LCMS (ESI) m / z: 230.2 [M+H] + .

[0766] Step 3: Synthesis of compound 64

[0767] In a dry single-mouth bottle, add substrate IM-1 (18 mg, 50.36 μmol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (13 mg, 137.36 μmol) and react at room temperature for 30 minutes. Then add N,N-diisopropylethylamine (9.7 mg, 75.05 μmol) and 64-2 (21 mg, 91.58 μmol) to the reaction solution and react at room temperature for 2 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 64 (6.2 mg, 11.51 μmol). LCMS (ESI) m / z: 539.2 [M+H] + ,HPLC method B:R T =5.59min,purity:92.7%. 1 H NMR (400 MHz, DMSO-d 6)δ8.80(s,1H),8.05–8.03(m,1H),7.79–7.77(m,2H),7.61(d,J=7.8Hz,3H),6.7 2(d,J=8.4Hz,2H),6.01–5.72(m,1H),5.72–5.54(m,1H),5.25–4.96(m,4H),4.83 (d,J=17.4Hz,1H),4.68(d,J=6.0Hz,2H),3.44(d,J=11.2Hz,2H),3.30(d,J=11. 2Hz, 2H), 3.18 (s, 2H), 2.93 (d, J = 6.0Hz, 2H), 1.55 (d, J = 8.4Hz, 1H), 1.47 (s, 6H).

[0768] Example 65: Synthesis of Compound 65

[0769]

[0770] Step 1: Synthesis of compound 65-1

[0771] Substrate 38-1 (87 mg, 396.83 μmol) was added to a dry single-mouth bottle, dissolved in acetone (3 mL), iodoethane (92.84 mg, 595.25 μmol) and potassium carbonate (83 mg, 1.19 mmol) were added, and the mixture was reacted at room temperature for 2 hours under a nitrogen atmosphere. LC-MS monitoring was performed. After the reaction was completed, the mixture was concentrated under reduced pressure, and the residue was purified by MPLC to obtain compound 65-1 (87 mg, 351.81 μmol). LCMS (ESI) m / z: 248.1 [M+H] + .

[0772] Step 2: Synthesis of compound 65-2

[0773] Add 65-1 (94 mg, 380.12 μmol) to a 50 mL round-bottom flask and dissolve in methanol (5 mL). Add a catalytic amount of palladium carbon (10 mg) and stir at room temperature for 1 hour under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 65-2 (80 mg, 368.14 μmol). LCMS (ESI) m / z: 218.2 [M+H] + .

[0774] Step 3: Synthesis of compound 65

[0775] In a dry single-mouth bottle, add substrate IM-1 (18 mg, 50.36 μmol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (13 mg, 137.36 μmol) and react at room temperature for 30 minutes. Then add N,N-diisopropylethylamine (9.7 mg, 75.05 μmol) and 65-2 (19.9 mg, 91.58 μmol) to the reaction solution and react at room temperature for 2 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 65 (6.2 mg, 11.77 μmol). LCMS (ESI) m / z: 527.2 [M+H] + ,HPLC method B:R T =6.27min,purity:76.2%. 1 HNMR (400 MHz, DMSO-d 6 )δ10.09(s,1H),8.80(s,1H),8.06–8.02(m,1H),7.76(d,J=8.0Hz,1H),7.60(d,J=7.6Hz,3H) ,6.70(d,J=8.4Hz,2H),5.74–5.55(m,1H),5.31(s,1H),5.04–4.94(m,1H),4.83(d,J=17.2Hz, 1H),4.68(d,J=6.0Hz,2H),3.67(d,J=5.6Hz,2H),3.42(d,J=11.2Hz,2H),3.26(d,J=11.2Hz,2 H),2.42(d,J=6.8Hz,1H),2.28(d,J=7.2Hz,2H),1.55(s,1H),1.46(s,6H),0.93–0.91(m,3H).

[0776] Example 66: Synthesis of Compound 66

[0777]

[0778] Step 1: Synthesis of compound 66-1

[0779] Add substrate 38-1 (80 mg, 364.90 μmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (3 mL), add 2-bromoethanol (68 mg, 544.16 μmol), potassium carbonate (99 mg, 716.35 μmol) and potassium iodide (5 mg, 30.12 μmol), and react at 100°C for 2 hours. LC-MS monitoring. After the reaction, the reaction solution was cooled to room temperature, extracted with ethyl acetate three times, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 66-1 (70 mg, 265.86 μmol). LCMS (ESI) m / z: 264.1 [M+H] + .

[0780] Step 2: Synthesis of compound 66-2

[0781] Add 66-1 (70 mg, 265.86 μmol) to a 50 mL round-bottom flask and dissolve in methanol (3 mL). Add a catalytic amount of palladium carbon (15 mg) and stir at room temperature for 3 hours under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 66-2 (35 mg, 150 μmol). LCMS (ESI) m / z: 234.2 [M+H] + .

[0782] Step 3: Synthesis of compound 66

[0783] Add substrate IM-6 (50 mg, 128.39 μmol) and tetrahydrofuran (4 mL) to a dry single-mouth bottle, stir and dissolve, then add N,N-diisopropylethylamine (82.97 mg, 641.97 μmol) and 66-2 (32 mg, 57.32 μmol), and react at room temperature for 4 hours. LC-MS monitoring. After the reaction is completed, the solvent is removed by distillation under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (basic) to obtain compound 66 (45 mg, 74.45 μmol). LCMS (ESI) m / z: 543.2 [M+H] + ,HPLC method B:R T =5.69min,purity:97.2%. 1H NMR(600MHz,Chloroform-d)δ8.81(s,1H),7.87-7.84(m,1H),7.76(d,J=7.8Hz,1H),7.47( s,2H),7.34(d,J=7.8Hz,1H),6.72(d,J=9.0Hz,2H),5.73-5.67(m,1H),5.04(d,J=10.2Hz,1 H),4.94(d,J=16.8Hz,1H),4.73(d,J=6.0Hz,2H),3.97(d,J=5.4Hz,2H),3.67-3.63(m,4H), 3.44(d,J=10.8Hz,2H),2.82-2.80(m,1H),2.64(s,2H),1.72(d,J=8.4Hz,1H),1.59(s,6H).

[0784] Example 67: Synthesis of Compound 67

[0785]

[0786] Step 1: Synthesis of compound 67-1

[0787] Add substrate 38-1 (80 mg, 364.90 μmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (3 mL), add 2-bromoethyl methyl ether (76 mg, 546.80 μmol), potassium carbonate (99 mg, 716.35 μmol) and potassium iodide (5 mg, 30.12 μmol), and react at 100°C for 6 hours. LC-MS monitoring. After the reaction, the reaction solution was cooled to room temperature, extracted with ethyl acetate three times, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 67-1 (75 mg, 270.45 μmol). LCMS (ESI) m / z: 264.1 [M+H] + .

[0788] Step 2: Synthesis of compound 67-2

[0789] Add 67-1 (75 mg, 270.45 μmol) to a 50 mL round-bottom flask and dissolve in methanol (3 mL). Add a catalytic amount of palladium carbon (15 mg) and stir at room temperature for 3 hours under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 67-2 (53 mg, 214.28 μmol). LCMS (ESI) m / z: 234.2 [M+H] + .

[0790] Step 3: Synthesis of compound 67

[0791] Add substrate IM-6 (50 mg, 128.39 μmol) and tetrahydrofuran (3 mL) to a dry single-mouth bottle, stir and dissolve, then add N,N-diisopropylethylamine (82.97 mg, 641.97 μmol) and 67-2 (53 mg, 214.28 μmol), and react at room temperature for 4 hours. LC-MS monitoring. After the reaction is completed, the solvent is removed by distillation under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (basic) to obtain compound 67 (40 mg, 67.11 μmol). LCMS (ESI) m / z: 557.3 [M+H] + ,HPLC method B:R T =6.43min,purity:93.4%. 1 H NMR(600MHz,Chloroform-d)δ8.82(s,1H),7.88-7.84(m,1H),7.77(d,J=8.4Hz,1H),7.46(s,2H), 7.34-7.32(m,1H),6.72(d,J=8.4Hz,2H),5.76-5.66(m,1H),5.06–5.03(m,1H),4.97–4.92(m,1H) ,4.74(d,J=6.0Hz,2H),3.95-3.93(m,2H),3.65(d,J=10.8Hz,2H),3.52-3.50(m,2H),3.39(d,J=1 0.8Hz, 2H), 3.35 (s, 3H), 2.83-2.81 (m, 1H), 2.66-2.64 (m, 2H), 1.68 (d, J = 8.4Hz, 1H), 1.59 (s, 6H).

[0792] Example 68: Synthesis of Compound 68

[0793]

[0794] Step 1: Synthesis of compound 68-1

[0795] In a dry single-mouth bottle, acetic acid (18 mg, 299.74 μmol) and N, N-diisopropylethylamine (42.61 mg, 329.72 μmol) were dissolved in tetrahydrofuran (2 mL), the temperature was lowered to 0 ° C, HATU (49.93 mg, 131.39 μmol) was added, and stirred for 10 minutes. After adding 38-1 (65.72 mg, 299.74 μmol), it was stirred for 30 minutes. After the reaction was completed, it was quenched with water, extracted with ethyl acetate three times, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by MPLC to obtain compound 68-1 (45 mg, 172.23 μmol). LCMS (ESI) m / z: 262.1 [M + H] + .

[0796] Step 2: Synthesis of compound 68-2

[0797] Add 68-1 (45 mg, 172.23 μmol) to a 50 mL round-bottom flask and dissolve in methanol (3 mL). Add a catalytic amount of palladium carbon (20 mg) and stir at room temperature for 1 hour under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 68-2 (29 mg, 214.28 μmol). LCMS (ESI) m / z: 236.2 [M+H] + .

[0798] Synthesis of compound 68

[0799] Add substrate IM-6 (50 mg, 128.39 μmol) and tetrahydrofuran (3 mL) to a dry single-mouth bottle, stir and dissolve, then add N,N-diisopropylethylamine (82.97 mg, 641.97 μmol) and 68-2 (29 mg, 125.38 μmol), and react at room temperature for 4 hours. LC-MS monitoring. After the reaction is completed, the solvent is removed by distillation under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (basic) to obtain compound 68 (51.3 mg, 89.10 μmol). LCMS (ESI) m / z: 541.3 [M+H] + ,HPLC method B:R T =6.04min,purity:93.9%. 1H NMR(600MHz,Chloroform-d)δ8.79(s,1H),7.87-7.83(m,1H),7.76(d,J=8.4Hz,1H) ,7.45(d,J=7.2Hz,2H),7.33(d,J=7.6Hz,1H),6.68(d,J=8.4Hz,2H),5.75-5.65(m,1 H),5.05–5.03(m,1H),4.94(d,J=17.4Hz,1H),4.74(d,J=6.0Hz,2H),4.61-4.54(m,2 H),4.09(brs,1H),3.97-3.95(m,1H),3.68-3.58(m,2H),3.43(d,J=9.0Hz,1H),2.78

[0800] -2.73(m,1H),1.94(s,3H),1.79(brs,1H),1.71(d,J=8.4Hz,1H),1.59(s,6H).

[0801] Example 69: Synthesis of Compound 69

[0802]

[0803] Step 1: Synthesis of compound 69-1

[0804] Add substrate 38-1 (241.29 mg, 1.10 mmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (5 mL), add methyl chloroformate (104 mg, 1.10 mmol) and potassium carbonate (194 mg, 1.40 mmol), and react at room temperature for 5 hours. Monitor by LC-MS. After the reaction, extract with ethyl acetate three times, dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain compound 69-1 (60 mg, 216.39 μmol). LCMS (ESI) m / z: 278.1 [M+H] + .

[0805] Step 2: Synthesis of compound 69-2

[0806] Add 69-1 (75 mg, 270.45 μmol) to a 50 mL round-bottom flask and dissolve in methanol (3 mL). Add a catalytic amount of palladium carbon (15 mg) and stir at room temperature for 1 hour under a hydrogen atmosphere. Monitor by LC-MS. After the reaction is completed, filter the reaction solution with diatomaceous earth, wash twice with methanol, and concentrate the filtrate under reduced pressure to obtain compound 69-2 (38 mg, 153.66 μmol). LCMS (ESI) m / z: 248.1 [M+H] + .

[0807] Step 3: Synthesis of compound 69

[0808] Add substrate IM-6 (50 mg, 128.39 μmol) and tetrahydrofuran (3 mL) to a dry single-mouth bottle, stir and dissolve, then add N,N-diisopropylethylamine (82.97 mg, 641.97 μmol) and 69-2 (38 mg, 153.66 μmol), and react at room temperature for 4 hours. LC-MS monitoring. After the reaction is completed, the solvent is removed by distillation under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (basic) to obtain compound 69 (35 mg, 62.88 μmol). LCMS (ESI) m / z: 557.2 [M+H] + ,HPLC method B:R T =7.13min,purity:98.7%. 1 H NMR(600MHz,Chloroform-d)δ8.79(s,1H),7.87-7.84(m,1H),7.77(d,J=8.4Hz,1H), 7.44(s,2H),7.33(d,J=7.2Hz,1H),6.68(d,J=8.4Hz,2H),5.75-5.65(m,1H),5.05–5 .02(m,1H),4.96–4.91(m,1H),4.74(d,J=6.0Hz,2H),4.39(d,J=5.4Hz,2H),3.93-3. 83(m,2H),3.65(s,3H),3.37(d,J=10.2Hz,2H),2.74-2.69(m,1H),1.60-1.59(m,7H).

[0809] Example 70: Synthesis of Compound 70

[0810]

[0811] Step 1: Synthesis of compound 70-2

[0812] In a dry single-mouth bottle, 70-1 (100 mg, 507.53 μmol) was dissolved in tetrahydrofuran (3 ml), the temperature was lowered to 0°C, NaH (14.62 mg, 609.04 μmol) was added, and the mixture was stirred at 0°C for 10 minutes. Ethyl iodide (94.99 mg, 609.04 μmol) was added, the temperature was raised to room temperature and stirred for 2 hours. LC-MS monitoring, after the reaction was completed, the mixture was quenched with water, extracted with ethyl acetate three times, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure, and the residue was purified by MPLC to obtain compound 70-2 (65 mg, 288.78 μmol). LCMS (ESI) m / z: 225.0 [M+H] + .

[0813] Step 2: Synthesis of compound 70-3

[0814] Dissolve 70-2 (65 mg, 288.78 μmol) in trifluoroacetic acid (1 ml) in a dry single-mouth bottle, add acetic anhydride (30.62 mg, 288.78 μmol, 0.5 mL), and heat to 70 ° C and stir for 4 hours. LC-MS monitoring, after the reaction is completed, cool to room temperature, concentrate under reduced pressure, and the residue is purified by MPLC to obtain compound 70-3 (40 mg, 149.74 μmol). LCMS (ESI) m / z: 267.0 [M+H] + .

[0815] Step 3: Synthesis of compound 70-4

[0816] In a dry microwave tube, under nitrogen protection, add substrate 70-3 (32.78 mg, 122.70 μmol), IM-1-4 (30 mg, 134.97 μmol), cuprous iodide (46.74 mg, 245.41 μmol), potassium carbonate (23.74 mg, 171.78 μmol) and N, N'-dimethylethylenediamine (43.27 mg, 490.81 μmol), and then add 1,4-dioxane (2 mL). Under nitrogen environment, heat the reaction at 110 ° C for 3 hours. LC-MS monitoring, after the reaction is completed, the reaction system is cooled to room temperature, concentrated under reduced pressure to remove the solvent, and the residue is purified by MPLC to obtain compound 70-4 (33 mg, 74.26 μmol). LCMS (ESI) m / z: 409.1 [M+H] + .

[0817] Step 4: Synthesis of compound 70

[0818] In a dry single-mouth bottle, add substrate 70-4 (59.71 mg, 146.17 μmol) and tetrahydrofuran (1.5 mL) to dissolve. Add m-chloroperbenzoic acid (45.40 mg, 263.10 μmol) and react at room temperature for 30 minutes. Then add N,N-diisopropylethylamine (94.45 mg, 730.83 μmol) and 1-5 (33.55 mg, 175.40 μmol) to the reaction solution and react at room temperature for 12 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (basic) to obtain compound 70 (23 mg, 40.40 μmol). LCMS (ESI) m / z: 552.2 [M+H] + ,HPLC method B:R T =6.81min,purity:97.6%. 1H NMR (600 MHz, DMSO-d 6 )δ10.14(s,1H),8.84(s,1H),8.64(d,,J=10.8Hz,2H),7.74(d,J=8.4Hz, 1H),7.56(s,2H),6.91-6.89(m,2H),5.71-5.66(m,1H),4.50–4.99(m,1H) ,4.85(d,J=17.4Hz,1H),4.66-4.59(m,2H),4.35–4.32(m,2H),3.09-3.0 8(m,4H),2.49(s,3H),2.48-2.46(m,4H),2.23(s,3H),1.47-1.45(m,3H).

[0819] Example 71: Synthesis of Compound 71

[0820]

[0821] Step 1: Synthesis of compound 71-1

[0822] Dissolve 70-1 (600 mg, 2.67 mmol) in 1,4-dioxane (1 ml) in a dry single-mouth bottle, add pyridinium tribromide (3.41 g, 10.66 mmol), and stir at room temperature for 16 hours. Monitor by LC-MS. After the reaction is completed, add water, extract with dichloromethane three times, dry the organic phase over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is purified by MPLC to obtain compound 71-1 (230 mg, 576.62 μmol). LCMS (ESI) m / z: 398.8 [M+H] + .

[0823] Step 2: Synthesis of compound 71-2

[0824] Dissolve 71-1 (230 mg, 576.62 μmol) in tetrahydrofuran (1.5 mL) in a dry single-mouth bottle, add zinc powder (45.57 mg, 696.93 μmol) and saturated ammonium chloride solution (1.5 mL), and stir at room temperature for 10 minutes. Monitor by LC-MS, filter with diatomaceous earth after the reaction, concentrate the filtrate under reduced pressure, and purify the residue by MPLC to obtain compound 71-2 (44 mg, 182.51 μmol). LCMS (ESI) m / z: 241.0 [M+H] + .

[0825] Step 3: Synthesis of compound 71-3

[0826] Dissolve 71-2 (250 mg, 1.16 mmol) in tetrahydrofuran (2 ml) in a dry single-mouth bottle, lower the temperature to 0°C, add NaH (83.30 mg, 3.47 mmol), and stir at 0°C for 10 minutes. Add iodomethane (492.7 mg, 3.47 mmol), warm to room temperature and stir for 6 hours. Monitor by LC-MS, quench with water after the reaction is completed, extract with ethyl acetate three times, dry the organic phase over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is purified by column chromatography to obtain compound 71-3 (210 mg, 912.64 μmol). LCMS (ESI) m / z: 269.0 [M+H] + .

[0827] Step 4: Synthesis of compound 71-4

[0828] In a dry microwave tube, under nitrogen protection, add substrate 71-3 (17.61 mg, 65.44 μmol), IM-1-4 (16 mg, 71.99 μmol), cuprous iodide (24.93 mg, 130.88 μmol), potassium carbonate (12.66 mg, 91.62 μmol) and N, N'-dimethylethylenediamine (23.07 mg, 261.77 μmol), and then add 1,4-dioxane (1 mL). Under nitrogen environment, heat the reaction at 110 ° C for 3 hours. LC-MS monitoring, after the reaction is completed, the reaction system is cooled to room temperature, concentrated under reduced pressure, and the residue is purified by MPLC to obtain compound 71-4 (2.01 mg, 4.91 μmol). LCMS (ESI) m / z: 411.2 [M+H] + .

[0829] Step 5: Synthesis of compound 71

[0830] In a dry single-mouth bottle, add substrate 71-4 (2.01 mg, 4.91 μmol) and add tetrahydrofuran (2 mL) to dissolve. Add m-chloroperbenzoic acid (1.52 mg, 8.83 μmol) and react at room temperature for 1 hour. Then add N,N-diisopropylethylamine (3.17 mg, 24.54 μmol, 4.27 μL) and 1-5 (938.70 μg, 4.91 μmol) to the reaction solution and react at room temperature for 8 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 71 (2.3 mg, 3.45 μmol). LCMS (ESI) m / z: 554.3 [M+H] + ,HPLC method B:R T =5.89min,purity:83.2%. 1HNMR(600MHz,CDCL3)δ8.77(s,1H),7.51(d,J=7.8Hz,1H),7.42(d,J=1.8Hz,2H) ,7.36(d,J=8.4Hz,1H),6.87-6.84(m,2H),5.70-5.63(m,1H),4.99(d,J=10.2Hz ,1H),4.92(d,J=16.8Hz,1H),4.63(d,J=5.4Hz,2H),3.80-3.77(m,2H),3.36-3. 24(m,4H),2.87-2.80(m,4H),2.49-2.45(m,3H),1.36(s,6H),1.23–1.21(m,3H).

[0831] Example 72: Synthesis of Compound 72

[0832]

[0833] Step 1: Synthesis of compound 72-21

[0834] Add substrate 72-1 (88 mg, 395.92 μmol) to a dry single-mouth bottle and dissolve in dichloromethane (20 mL). Add IM-1-4 (170 mg, 764.85 μmol), pyridine (0.4 mL), copper acetate (146 mg, 803.83 μmol), and stir at 30°C for 24 hours. Monitor by LC-MS. After the reaction is completed, concentrate under reduced pressure, and the residue is purified by MPLC to obtain compound 72-2 (28 mg, 79.00 μmol). LCMS (ESI) m / z: 354.95 [M+H] + .

[0835] Step 2: Synthesis of compound 72

[0836] In a dry single-mouth bottle, add substrate 72-2 (28 mg, 79.00 μmol) and tetrahydrofuran (2 mL) to dissolve. Add m-chloroperbenzoic acid (27.26 mg, 157.99 μmol) and react at room temperature for 1 hour. Then add N,N-diisopropylethylamine (16 mg, 83.65 μmol) and 1-5 (938.70 μg, 4.91 μmol) to the reaction solution and react at room temperature for 2 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 72 (9 mg, 17.72 μmol). LCMS (ESI) m / z: 498.1 [M+H] + ,HPLC method B:R T =8.01min,purity:98%. 1HNMR(400MHz,Chloroform-d)δ8.84(s,1H),7.85-7.81(m,2H),7.48–7.41(m,5H),6.86(d,J=8.4Hz,2H ),5.79-5.72(m,1H),5.18–5.09(m,2H),4.47(d,J=6.0Hz,2H),3.32(s,4H),2.86(s,4H),2.55(s,3H).

[0837] Example 73: Synthesis of Compound 73

[0838]

[0839] Step 1: Synthesis of compound 73-2

[0840] Add substrate 73-1 (0.5 g, 3.28 mmol) to a dry single-mouth bottle and dissolve in acetonitrile (5 mL). Add pyridine (1.5 mL) and Selectfluor reagent (1.05 g, 2.95 mmol) and stir at 15°C for 16 hours. Monitor by LC-MS. After the reaction is completed, add water and extract with ethyl acetate three times. The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by prep-HPLC to obtain compound 73-2 (112 mg, 656.62 μmol). LCMS (ESI) m / z: 171.0 [M+H] + .

[0841] Step 2: Synthesis of compound 73-3

[0842] In a dry single-mouth bottle, 73-2 (0.112 g, 656.62 μmol) was dissolved in N, N-dimethylformamide (3 ml), the temperature was lowered to 0°C, NaH (39.40 mg, 984.93 μmol, 60% purity) was added, and the mixture was stirred at 0°C for 30 minutes. Ethyl iodide (153.65 mg, 984.93 μmol, 78.79 μL) was added, the mixture was warmed to room temperature and stirred for 16 hours. LC-MS monitoring, after the reaction was completed, the mixture was quenched with saturated ammonium chloride solution, extracted with ethyl acetate three times, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure, and the residue was purified by column chromatography to obtain compound 73-3 (120 mg, 604.16 μmol). LCMS (ESI) m / z: 199.2 [M+H] + Step 3: Synthesis of compound 73-4

[0843] In a dry microwave tube, under nitrogen protection, add substrate 74-3 (0.04 g, 201.39 μmol), IM-1-4 (44.76 mg, 201.39 μmol), cuprous iodide (76.71 mg, 402.77 μmol), sodium iodide (60.37 mg, 402.77 μmol), potassium carbonate (69.58 mg, 503.46 μmol) and N, N'-dimethyl-1,2-cyclohexanediamine (114.58 mg, 805.54 μmol), and then add anisole (1.5 mL). Under nitrogen environment, microwave heating at 130 ° C for 4 hours. LC-MS monitoring, after the reaction is completed, the reaction system is cooled to room temperature, water and ammonia (1 mL) are added, and ethyl acetate is extracted three times. The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by column chromatography to obtain compound 73-4 (20 mg, 52.03 μmol). LCMS (ESI) m / z: 385.0 [M+H] + .

[0844] Step 4: Synthesis of compound 73

[0845] In a dry single-mouth bottle, add substrate 73-4 (20 mg, 52.03 μmol) and tetrahydrofuran (1.5 mL) to dissolve. Add m-chloroperbenzoic acid (19.01 mg, 93.65 μmol, 85% purity) and react at room temperature for 1 hour. Then add N,N-diisopropylethylamine (67.24 mg, 520.25 μmol, 90.62 μL) and 1-5 (11.94 mg, 62.43 μmol) to the reaction solution and react at room temperature for 5 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 73 (20.7 mg, 37.39 μmol). LCMS (ESI) m / z: 528.3 [M+H] + ,HPLC method B:R T =8.09min,purity:95.3%. 1 H NMR (600 MHz, DMSO-d 6)δ10.16(s,1H),8.84(s,1H),8.32(s,1H),7.67(s,1H),7.65(d,J=8.4 Hz,1H),7.61(s,2H),6.92(d,J=8.4Hz,2H),5.74–5.68(m,1H),5.00(d, J=10.2Hz,1H),4.86(d,J=17.4Hz,1H),4.65(s,2H),4.26–4.22(m,2H) ,3.12–3.09(m,4H),2.47–2.45(m,4H),2.22(s,3H),1.39–1.36(m,3H).

[0846] Example 74: Synthesis of Compound 74

[0847]

[0848] Step 1: Synthesis of compound 74-2

[0849] Add substrate 74-1 (1.01 g, 3.99 mmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (30 mL), add methyl propylene oxide (345.57 mg, 4.79 mmol), potassium carbonate (1.66 g, 11.98 mmol), and raise the temperature to 120°C for 8 hours. Monitor by LC-MS. After the reaction is completed, cool to room temperature, extract with ethyl acetate three times, dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is purified by column chromatography to obtain compound 74-2 (680 mg, 2.09 mmol). LCMS (ESI) m / z: 311.9 [M+H] + .

[0850] Step 2: Synthesis of compound 74-3

[0851] In a dry single-mouth bottle, 74-2 (98 mg, 301.54 μmol) was dissolved in tetrahydrofuran (4 ml), the temperature was lowered to 0°C, NaH (24.12 mg, 603.08 μmol, 60% purity) was added in portions, the temperature was raised to 60°C and stirred for 30 hours. LC-MS monitoring, after the reaction was completed, the temperature was lowered to room temperature, quenched with saturated aqueous ammonium chloride solution, extracted with ethyl acetate three times, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure, and the residue was purified by MPLC to obtain compound 74-3 (48 mg, 196.65 μmol). LCMS (ESI) m / z: 244.0 [M+H] + .

[0852] Step 3: Synthesis of compound 74-4

[0853] In a dry microwave tube, under nitrogen protection, add substrate 74-3 (47 mg, 192.56 μmol), IM-1-4 (31.77 mg, 142.96 μmol), cuprous iodide (36.67 mg, 192.56 μmol), potassium carbonate (38.30 mg, 277.14 μmol) and N, N'-dimethylethylenediamine (33.95 mg, 385.11 μmol), and then add 1,4-dioxane (3 mL). Under nitrogen environment, microwave heating at 120 ° C for 4 hours. LC-MS monitoring, after the reaction is completed, the reaction system is cooled to room temperature, concentrated under reduced pressure, and the residue is purified by column chromatography to obtain compound 74-4 (12 mg, 31.13 μmol). LCMS (ESI) m / z: 386.1 [M+H] + .

[0854] Step 4: Synthesis of compound 74

[0855] In a dry single-mouth bottle, add substrate 74-4 (12 mg, 31.13 μmol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (21.49 mg, 124.53 μmol) and react at room temperature for 0.5 hours. Then add N,N-diisopropylethylamine (20.22 mg, 156.46 μmol, 27.25 μL) and 1-5 (20 mg, 104.56 μmol) to the reaction solution and react at room temperature for 8 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 74 (7.2 mg, 12.56 μmol). LCMS (ESI) m / z: 529.2 [M+H] + ,HPLC method B:R T =7.08min,purity:92.2%. 1 H NMR(600MHz,Chloroform-d)δ8.80(s,1H),7.47(d,J=7.8Hz,2H),7.37-7.30(m,2H),6.90(d,J=8.4Hz,2H),5.74-5.68(m,1H), 5.05(d,J=10.2Hz,1H),4.98(d,J=16.8Hz,1H),4.69(s,2H),4.11(s,2H),3.24(s,4H),2.69(s,4H),2.43(s,3H),1.41(s,6H).

[0856] Example 75: Synthesis of Compound 75

[0857]

[0858] Step 1: Synthesis of compound 75-2

[0859] Add substrate 75-1 (965 mg, 5.00 mmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (20 mL), add iodoethane (383 mg, 4.70 mmol), N,N-diisopropylethylamine (1.29 g, 10.00 mmol), and react at room temperature for 1 hour. Monitor by TLC. After the reaction is completed, extract with ethyl acetate three times, dry the organic phase with anhydrous sodium sulfate, filter, concentrate under reduced pressure, and purify the residue by column chromatography to obtain compound 75-2 (650 mg, 3.22 mmol). LCMS (ESI) m / z: 202.0 [M+H] + .

[0860] Step 2: Synthesis of compound 75-3

[0861] Dissolve 75-2 (308 mg, 1.53 mmol) in tetrahydrofuran (5 mL) in a dry single-mouth bottle, add zinc powder (1.6 g, 24.62 mmol) and ammonium chloride (642 mg, 12.00 mmol), and stir at 70 °C for 3 hours. Monitor by LC-MS, filter with diatomaceous earth after the reaction, concentrate the filtrate under reduced pressure, and purify the residue by MPLC to obtain compound 75-3 (248 mg, 1.44 mmol). LCMS (ESI) m / z: 172.1 [M+H] + .

[0862] Step 3: Synthesis of compound 75-4

[0863] Dissolve 75-3 (248 mg, 1.44 mmol) in triethyl orthoacetate (4 ml) in a dry single-mouth bottle and stir at 100°C for 1 hour. Add acetic acid (1.05 g, 17.47 mmol, 1 mL) and continue stirring for 8 hours. Monitor by LC-MS. After the reaction is completed, cool to room temperature, concentrate under reduced pressure, extract with ethyl acetate 3 times, dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is purified by MPLC to obtain compound 75-4 (220 mg, 1.12 mmol). LCMS (ESI) m / z: 196.1 [M+H] + Step 4: Synthesis of compound 75-5

[0864] In a dry microwave tube, under nitrogen protection, add substrate 75-4 (64 mg, 327.12 μmol), IM-1-4 (72.71 mg, 327.12 μmol), cuprous iodide (124.60 mg, 654.23 μmol), sodium iodide (98.06 mg, 654.23 μmol), potassium carbonate (38.30 mg, 277.14 μmol) and N, N'-dimethyl-1,2-cyclohexanediamine (186.12 mg, 1.31 mmol), and then add anisole (2 mL). Under nitrogen environment, microwave heating at 130 ° C for 10 hours. LCMS monitoring, after the reaction is completed, the reaction system is cooled to room temperature, water and ammonia (1 mL) are added, and ethyl acetate is extracted three times. The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by column chromatography to obtain compound 75-5 (15 mg, 39.32 μmol). LCMS (ESI) m / z: 382.2 [M+H] + .

[0865] Step 5: Synthesis of compound 75

[0866] In a dry single-mouth bottle, add substrate 75-5 (15 mg, 39.32 μmol) and tetrahydrofuran (2 mL) to dissolve. Add m-chloroperbenzoic acid (20.36 mg, 117.97 μmol) and react at room temperature for 1 hour. Then add N,N-diisopropylethylamine (25.36 mg, 196.25 μmol, 34.18 μL) and 1-5 (18.77 mg, 98.12 μmol) to the reaction solution and react at room temperature for 8 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 75 (3.2 mg, 5.12 μmol). LCMS (ESI) m / z: 525.2 [M+H] + ,HPLC method B:R T =6.16min,purity:84.0%. 1 H NMR(600MHz,Chloroform-d)δ8.84(s,1H),8.07(d,J=7.8Hz,1H),7.66(d,J=7.2Hz,1H),7.46(d,J=8.4Hz,2H),6.88(d,J=8.4Hz,2H),5.75-5.68(m,1H), 5.00(d,J=10.2Hz,1H),4.89(d,J=17.4Hz,1H),4.73(d,J=3.6Hz,2H),4.32– 4.28(m,2H),3.25(s,4H),2.71-2.68(m,7H),2.45(s,3H),1.47-1.42(m,3H).

[0867] Example 76: Synthesis of Compound 76

[0868]

[0869] Step 1: Synthesis of compound 76

[0870] In a dry single-mouth bottle, add substrate 51-2 (26 mg, 70.95 μmol) and tetrahydrofuran (2 mL) to dissolve. Add m-chloroperbenzoic acid (35 mg, 202.82 μmol) and react at 50°C for 2 hours. Then add N,N-diisopropylethylamine (45.62 mg, 353.00 μmol) and IM-2 (22 mg, 108.22 μmol) to the reaction solution and react at room temperature for 8 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (basic) to obtain compound 76 (15 mg, 28.76 μmol). LCMS (ESI) m / z: 522.3 [M+H] + ,HPLC method B:R T =7.68min,purity:94.4%. 1 H NMR(600MHz,Chloroform-d)δ8.83(s,1H),8.03(d,J=7.8Hz,1H),7.53-7.43(m,3H),7.2 8(d,J=3.6Hz,1H),6.54–6.52(m,3H),5.75-5.69(m,1H),5.00(d,J=10.2Hz,1H),4.90(( d,J=17.4Hz,1H),4.76(s,2H),4.34-4.28(m,3H),3.78(s,1H),3.51–3.45(m,2H),3.21( s,1H),2.84(s,1H),2.53(s,3H),2.21(s,1H),2.06(d,J=9.6Hz,1H),1.51–1.46(m,3H).

[0871] Example 77: Synthesis of Compound 77

[0872]

[0873] Step 1: Synthesis of compound 77

[0874] In a dry single-mouth bottle, add substrate 51-2 (26 mg, 70.95 μmol) and tetrahydrofuran (2 mL) to dissolve. Add m-chloroperbenzoic acid (35 mg, 202.82 μmol) and react at 55°C for 6 hours. Then add N,N-diisopropylethylamine (45.62 mg, 353.00 μmol) and 77-1 (17 mg, 89.34 μmol) to the reaction solution and react at room temperature for 8 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (basic) to obtain compound 77 (5 mg, 9.83 μmol). LCMS (ESI) m / z: 509.6 [M+H] + ,HPLC method B:R T =8.50min,purity:98.2%. 1 HNMR (600 MHz, DMSO-d 6 )δ10.18(s,1H),8.87(s,1H),8.23(d,J=7.8Hz,1H),7.67(d,J=3.0Hz,3H),7.53(d,J=7. 8Hz,1H),7.16(d,J=8.4Hz,2H),6.58(d,J=3.6Hz,1H),5.74-5.68(m,1H),5.00(d,J=7.2 Hz,1H),4.86(d,J=16.8Hz,1H),4.61(s,2H),4.30–4.26(m,2H),2.86(d,J=11.4Hz,2H), 2.41-2.39(m,1H),2.19(s,3H),1.97-1.93(m,2H),1.71-1.62(m,4H),1.41–1.38(m,3H).

[0875] Example 78: Synthesis of Compound 78

[0876]

[0877] Step 1: Synthesis of compound 78-1

[0878] 3,4-difluoronitrobenzene (159 mg, 999.43 μmol) was dissolved in dimethyl sulfoxide (3 mL) in a dry single-mouth bottle, and 27-1 (237.78 mg, 1.20 mmol) and potassium carbonate (137.92 mg, 999.43 μmol) were added in sequence. The reaction mixture was stirred at 100 °C overnight. After the reaction was completed, it was diluted with water and extracted with ethyl acetate three times. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound 78-1 (302 mg, 890 μmol). LCMS (ESI) m / z: 338.2 [M+H]+ .

[0879] Step 2: Synthesis of compound 78-2

[0880] Add substrate 78-1 (337.35 mg, 1 mmol) to a dry single-mouth bottle and dissolve it in dichloromethane (2 mL). Slowly add trifluoroacetic acid (1.54 g, 13.51 mmol, 1 mL) and react at room temperature for 0.5 hours, and monitor with LC-MS. After the reaction is completed, remove the solvent by distillation under reduced pressure to obtain compound 78-2 (223 mg, 0.94 mmol). LCMS (ESI) m / z: 238.1 [M+H] + .

[0881] Step 3: Synthesis of compound 78-3

[0882] Add substrate 78-2 (223 mg, 0.94 mmol) dissolved in 1,2-dichloroethane (3 mL) to a dry single-mouth bottle, add formaldehyde (150.13 mg, 5.00 mmol), and stir at room temperature for 10 minutes. Add sodium triacetoxyborohydride (1.06 g, 5.00 mmol) and stir at room temperature for 1 hour. After the reaction is completed, cool to room temperature, dilute with water, extract three times with dichloromethane, dry the organic phase over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is purified by column chromatography to obtain compound 78-3 (198 mg, 0.78 mmol). LCMS (ESI) m / z: 252.1 [M+H] + .

[0883] Step 4: Synthesis of compound 78-4

[0884] In a dry single-mouth bottle, 78-3 (251.26 mg, 1 mmol) was dissolved in tetrahydrofuran (10 mL) and acetic acid (0.5 mL), and zinc powder (1.95 g, 30.00 mmol) was added and stirred at room temperature for 30 minutes. LC-MS monitoring, after the reaction was completed, diatomaceous earth was used for filtration, and the filtrate was concentrated under reduced pressure to obtain compound 78-4 (200 mg, 903.86 μmol). LCMS (ESI) m / z: 222.1 [M+H] + .

[0885] Step 5: Synthesis of Compound 78

[0886] In a dry single-mouth bottle, add substrate IM-1 (42.27 mg, 118.27 μmol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (30.61 mg, 177.40 μmol) and react at room temperature for 30 minutes. Then add N, N-diisopropylethylamine (76.43 mg, 591.35 μmol) and 78-4 (27 mg, 118.27 μmol) to the reaction solution and react at room temperature for 4 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 78 (2 mg, 3.72 μmol). LCMS (ESI) m / z: 531.7 [M+H] + .HPLC:R T =7.53min,purity:85.0%. 1 HNMR (600 MHz, DMSO-d 6 )δ10.28(s,1H),8.85(s,1H),8.002-7.99(m,1H),7.76(d,J=7.8Hz,1H),7.71-7.67(m,1H ),7.63(d,J=7.8Hz,1H),7.34(d,J=8.4Hz,1H),6.95-6.91(m,1H),5.70-5.64(m,1H),5.3 4(s,1H),5.00(d,J=10.2Hz,1H),4.83(d,J=17.4Hz,1H),4.68(d,J=6.0Hz,2H),3.63(d,J =11.1Hz,2H),3.50(d,J=5.9Hz,2H),3.43(s,2H),3.17(s,2H),2.04(s,3H),1.46(s,6H).

[0887] Example 79: Synthesis of Compound 79

[0888]

[0889] Step 1: Synthesis of compound 79-1

[0890] In a dry single-mouth bottle, 3,4-difluoronitrobenzene (354.16 mg, 2 mmol) was dissolved in dimethyl sulfoxide (5 mL), and 27-1 (475.83 mg, 2.40 mmol) and potassium carbonate (276.00 mg, 2.00 mmol) were added in sequence. The reaction mixture was stirred at 100 ° C for 1 hour. After the reaction was completed, it was diluted with water and extracted with ethyl acetate three times. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain compound 79-1 (635 mg, 1.79 mmol). LCMS (ESI) m / z: 356.1 [M+H]+ .

[0891] Step 2: Synthesis of compound 79-2

[0892] Add substrate 79-1 (635 mg, 1.79 mmol) to a dry single-mouth bottle and dissolve it in dichloromethane (5 mL). Slowly add trifluoroacetic acid (2 mL) and react at room temperature for 0.5 hours, and monitor with LC-MS. After the reaction is completed, remove the solvent by distillation under reduced pressure to obtain compound 79-2 (300 mg, 1.18 mmol). LCMS (ESI) m / z: 256.1 [M+H] + .

[0893] Step 3: Synthesis of compound 79-3

[0894] Add substrate 79-2 (150 mg, 587.73 μmol) to a dry single-mouth bottle and dissolve it in 1,2-dichloroethane (2 mL), add formaldehyde (88.24 mg, 2.94 mmol), and stir at room temperature for 10 minutes. Add sodium triacetoxyborohydride (622.81 mg, 2.94 mmol) and stir at room temperature for 1 hour. After the reaction is completed, cool to room temperature, dilute with water, extract three times with dichloromethane, dry the organic phase over anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain compound 79-3 (131 mg, 487 μmol). LCMS (ESI) m / z: 270.1 [M+H] + .

[0895] Step 4: Synthesis of compound 79-4

[0896] In a dry single-mouth bottle, 79-3 (131 mg, 487 μmol) was dissolved in tetrahydrofuran (3 mL) and acetic acid (0.2 mL), and zinc powder (0.96 g, 14.61 mmol) was added and stirred at room temperature for 30 minutes. LC-MS monitoring, after the reaction was completed, the filtrate was filtered with diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain compound 79-4 (123 mg, 515 μmol). LCMS (ESI) m / z: 240.1 [M+H] + .

[0897] Step 5: Synthesis of compound 79

[0898] In a dry single-mouth bottle, add substrate IM-1 (15 mg, 42.20 μmol) and tetrahydrofuran (1 mL) to dissolve. Add m-chloroperbenzoic acid (14.09 mg, 84.40 μmol) and react at room temperature for 30 minutes. Then add N,N-diisopropylethylamine (27.27 mg, 211.00 μmol) and 79-4 (10.10 mg, 42.20 μmol) to the reaction solution and react at room temperature for 3 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (alkaline) to obtain compound 79 (1.5 mg, 2.48 μmol). LCMS (ESI) m / z: 549.3 [M+H] + .HPLC:R T =7.95min,purity:90.6%. 1 H NMR (600 MHz, DMSO-d 6 )δ10.51(s,1H),8.95(s,1H),8.03-7.99(m,1H),7.75(d,J=12.0Hz,1H),7.67(d,J=12.0Hz,1H),7.55(d,J =7.8Hz,1H),5.71-5.64(m,1H),5.70-5.64(m,1H),5.35(s,1H),5.01-4.98(m,1H),4.84(d,J=1.8Hz,1H), 4.80(d,J=1.8Hz,1H),4.69(d,J=8.0Hz,1H),3.62(d,J=15.6Hz,2H),3.17(d,J=10.2Hz,2H),3.05(d,J=10 .2Hz,2H),2.67(d,J=2.4Hz,1H),2.33(d,J=3.0Hz,1H),2.21(s,3H),1.80(d,J=12.0Hz,1H),1.46(s,6H).

[0899] Example 80: Synthesis of Compound 80

[0900]

[0901] Step 1: Synthesis of compound 80-2

[0902] Add substrate 80-1 (581.98 mg, 3.02 mmol) to a dry single-mouth bottle, dissolve in N,N-dimethylformamide (10 mL), add dimethyl malonate (436 mg, 3.30 mmol), cesium carbonate (3 g, 9.05 mmol), and raise the temperature to 90°C for 2 hours. Monitor by LC-MS. After the reaction is completed, cool to room temperature, extract with ethyl acetate 3 times, dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is purified by column chromatography to obtain compound 80-2 (753 mg, 2.61 mmol). LCMS (ESI) m / z: 289.1 [M+H] + .

[0903] Step 2: Synthesis of compound 80-3

[0904] In a dry single-mouth bottle, 80-2 (753 mg, 2.61 mmol) was dissolved in ethanol (10 mL) and acetic acid (2 mL) solution, iron powder (582.80 mg, 10.44 mmol) was added, and the temperature was raised to 80°C for 2 hours. LC-MS monitoring, after the reaction was completed, diatomaceous earth was used for filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by MPLC to obtain compound 80-3 (417 mg, 1.84 mmol). LCMS (ESI) m / z: 227.1 [M+H] + .

[0905] Step 3: Synthesis of compound 80-4

[0906] Add substrate 80-3 (45 mg, 198.57 μmol) to a dry single-mouth bottle, dissolve in acetonitrile (10 mL), add iodomethane (0.22 g, 1.99 mmol), potassium carbonate (111 mg, 794.29 μmol), and react at 60°C for 8 hours. Monitor by LC-MS. After the reaction, cool the reaction solution to room temperature, add water, extract with ethyl acetate three times, dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by column chromatography to obtain compound 80-4 (15 mg, 58.90 μmol). LCMS (ESI) m / z: 255.1 [M+H] + .

[0907] Step 4: Synthesis of compound 80-5

[0908] In a dry microwave tube, under nitrogen protection, add substrate 80-4 (15 mg, 58.90 μmol), IM-1-4 (13.09 mg, 58.90 μmol), cuprous iodide (22.43 mg, 117.80 μmol), sodium iodide (17.66 mg, 117.80 μmol), potassium carbonate (25 mg, 176.70 μmol) and N, N'-dimethyl-1,2-cyclohexanediamine (32.2 mg, 235.6 μmol), and then add anisole (1.5 mL). Under nitrogen environment, microwave heating at 130 ° C for 4 hours. LCMS monitoring, after the reaction is completed, the reaction system is cooled to room temperature, water and ammonia (1 mL) are added, and ethyl acetate is extracted three times. The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by column chromatography to obtain compound 80-5 (10.5 mg, 23.84 μmol). LCMS (ESI) m / z: 441.1 [M+H] + .

[0909] Step 5: Synthesis of compound 80

[0910] In a dry single-mouth bottle, add substrate 80-5 (30 mg, 68.11 μmol) and tetrahydrofuran (1.5 mL) to dissolve. Add m-chloroperbenzoic acid (26.39 mg, 152.92 μmol) and react at room temperature for 1 hour. Then add N,N-diisopropylethylamine (19.76 mg, 152.92 μmol) and 1-5 (19.5 mg, 101.95 μmol) to the reaction solution and react at room temperature for 3 hours. LC-MS monitoring, after the reaction is completed, concentrate under reduced pressure, and purify the residue by preparative high performance liquid chromatography (basic) to obtain compound 80 (5.7 mg, 9.77 μmol). LCMS (ESI) m / z: 584.2 [M+H] + .HPLC:R T =6.37min,purity:98.8%. 1 HNMR (600 MHz, DMSO-d 6 )δ10.14(s,1H),8.82(s,1H),7.80(d,J=7.8Hz,2H),7.67–7.39(m,2H),6.92(d,J=8.4Hz,2H),5.66–5.63(m,1H),4.99(d,J=10.2 Hz,1H),4.81(d,J=17.4Hz,1H),4.46(s,2H),3.59(s,3H),3.28(s,3H),3.10(s,4H),2.48–2.43(m,4H),2.23(s,3H),1.58(s,3H).

[0911] Biological evaluation test:

[0912] Unless otherwise specified, compound AZD1775 was used as a control in some biological evaluation experiments in this part of the examples. The structural information of AZD1775 (CAS No.: 955365-80-7) is as follows:

[0913]

[0914] Experimental Example 1: Evaluation of the effect of compounds on the binding of WEE1 protein to Tracer 178 by TR-FRET method

[0915] First, prepare compound solutions with different concentration gradients. Dissolve the compound in DMSO and dilute the compound 4 times for a total of 10 dose points, set 2 parallel replicates for each concentration, add DMSO as a positive control (maximum signal control) and a negative control (minimum signal control), and ensure that the final DMSO content in each reaction well is 0.25%.

[0916] The compound was added to a 384-well plate using ECHO665, and then Wee1 protein solution was added. The protein was prepared in buffer (50mM HEPES pH 7.5, 10mM MgCl2, 1mM EGTA, 0.01% Brij-35) WEE1 (Thermo Fisher, Cat#PR7373A), and the final protein concentration was 15nM). The reaction substrate Tracer 178 (Invitrogen, PV5593) and MAbAnti-GST-Eu crypate (Cisbio, 61GSTKLA) were added to a 384-well plate (Corning, cat#3574), and the plate was centrifuged at 1000rpm for 1min. The 384-well reaction plate was placed in a constant temperature shaker and incubated for 60min at 25°C and 300rpm. Tracer 178 and MAbAnti-GST-Eu crypate were prepared in buffer (50mM HEPES pH 7.5, 10mM MgCl2, 1mM EGTA, 0.01% Brij-35), and the final reaction concentration of Tracer178 was 50nM, and the final concentration of MAb Anti-GST-Eu crypate was 2nM. The negative control (minimum signal control) used an equal amount of buffer instead of protein solution.

[0917] After incubation, the reading was performed using BMG PHERAStar (using 337 nm wavelength as excitation light and 620 nm and 665 nm wavelengths as emission light to read the fluorescence signal value). The fluorescence signal ratio was calculated: 665 / 620*1000 as the final enzyme activity signal value, and the TR-FRET signal obtained by the positive control (maximum signal control) and the negative control (minimum signal control) was normalized to obtain the inhibition rate of compounds at different concentrations. The IC50 of the compound's inhibition of enzyme activity was then calculated using GraphPad Prism 6 in the log(inhibitor) vs.response–Variable slope mode. The fitting equation is: Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*HillSlope)), where Y represents the known percentage of residual enzyme activity and X represents the concentration of the known compound after Log.

[0918] The Wee1 inhibitory activity of the example compounds was tested according to the above method. The test results are shown in Table 1, where the IC 50 The descriptions are classified as follows:

[0919] “-” indicates IC 50 Measured value 10 μM;

[0920] "+" indicates IC 50 The measured value is less than or equal to 10 μM and greater than 1 μM;

[0921] "++" indicates IC 50 The measured value is less than or equal to 1μM and greater than 100nM;

[0922] "+++" indicates IC 50 The measured value is less than or equal to 100nM and greater than 10nM;

[0923] "++++" indicates IC 50 The measured value is less than or equal to 10nM and greater than 5nM;

[0924] "++++++" indicates IC 50 The measured value was less than or equal to 5 nM.

[0925] Table 1. Inhibitory activity of the compounds of the present invention on Wee1 kinase

[0926] Compound No. <![CDATA[IC 50 / nM]]> Compound No. <![CDATA[IC 50 / nM]]> Compound No. <![CDATA[IC 50 / nM]]> AZD1775 ++++ Compound 1 ++++ Compound 3 +++++ Compound 4 +++++ Compound 5 +++++ Compound 6 +++++ Compound 7 +++++ Compound 8 ++++ Compound 9 +++++ Compound 10 +++++ Compound 11 ++++ Compound 12 ++++ Compound 13 ++++ Compound 14 ++++ Compound 15 ++++ Compound 16 +++ Compound 17 ++++ Compound 18 ++++ Compound 19 +++++ Compound 20 ++++ Compound 21 +++ Compound 22 +++ Compound 23 +++ Compound 24 +++ Compound 25 ++++ Compound 27 ++++ Compound 28 ++++ Compound 29 ++++ Compound 30 +++++ Compound 31 +++ Compound 32 +++ Compound 33 +++ Compound 34 +++ Compound 35 +++ Compound 36 +++ Compound 37 +++ Compound 38 +++ Compound 39 +++++ Compound 41 +++ Compound 42 +++ Compound 43 +++ Compound 44 ++++ Compound 45 +++ Compound 46 +++ Compound 47 +++++ Compound 47a +++ Compound 47b +++ Compound 48 +++++ Compound 49 +++++ Compound 50 ++ Compound 51 +++++ Compound 52 +++ Compound 53 ++++ Compound 54 ++++ Compound 55 +++ Compound 56 +++ Compound 57 +++ Compound 58 ++++ Compound 60 ++++ Compound 61 +++++ Compound 62 ++++ Compound 63 + Compound 64 +++ Compound 65 ++++ Compound 66 ++++ Compound 67 ++++ Compound 68 +++ Compound 69 +++ Compound 70 ++++ Compound 71 +++ Compound 72 +++ Compound 73 +++ Compound 74 +++ Compound 75 +++ Compound 76 +++ Compound 77 ++++ Compound 78 +++ Compound 79 +++

[0927] The above experiments show that the compounds disclosed in the present invention have obvious inhibitory activity on Wee1 kinase.

[0928] Experimental Example 2: Evaluation of cell proliferation inhibitory activity

[0929] 1. The antiproliferative activity of the compounds on H1299 cells was evaluated by the Cell Titer-Glo method.

[0930] H1299 (ATCC, cat#HTB-96) cells were seeded in a 384-well plate (Corning, 3707) at 750 cells / well in culture medium (90% 1640 (Hyclone, cat#16600082) + 10% FBS (Corning, Cat#35-081-CV) + 100ug / ml Normocin (InvivoGen, cat#Ant-nr-2)) and incubated overnight at 37°C, 5% CO2 to allow them to adhere.

[0931] Prepare compound solutions with different concentration gradients. Dissolve DMSO to a concentration of 10mM test compound and 10mM reference compound AZD1775, and dilute the compound in the culture medium in series, with a total of 9 dose points, and set 2 parallel replicates for each concentration. The cell growth group without compound addition serves as a positive control (maximum signal control), and the culture medium serves as a negative control (minimum signal control), while ensuring that the final DMSO content in each reaction well is 0.2%. After removing the culture medium from the 384-well plate, transfer 25ul of the prepared compounds of different concentrations into the well plate, and incubate the compounds and cells in a cell culture incubator at 37°C, 5% CO2 for 3 days.

[0932] Take the 384-well plate out of the cell culture incubator and let it equilibrate to room temperature for 1 hour. Then add 25ul of Cell Titer-Glo detection reagent to each reaction well. After shaking for 2 minutes, incubate at room temperature for 10 minutes and read the results (Luminescence) using BMG PHERAStar. Calculate the inhibition rate based on the luminescence signal: first calculate the average value of the positive control (maximum signal control) and the negative control (minimum signal control).

[0933] by To calculate the inhibition rate of different concentrations of compounds on cells. The IC50 of the compound's inhibition of cell activity was calculated by fitting in the log(inhibitor) vs.response–Variableslope mode using GraphPad Prism 6. The fitting equation is: Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*HillSlope)), where Y represents the inhibition rate and X represents the concentration of the known compound after Log.

[0934] 2. The antiproliferative activity of the compounds on MIA PaCa2 cells was evaluated by the Cell Titer-Glo method.

[0935] MIA PACA2 (ATCC, cat#CRL-1420) cells were seeded in 384-well plates (Corning, 3707) at a rate of 750 cells / well in culture medium (90% 1640 (Hyclone, cat#16600082) + 10% FBS (Corning, Cat#35-081-CV) + 100ug / ml Normocin (InvivoGen, cat#Ant-nr-2)) and incubated overnight at 37°C, 5% CO2 to allow them to adhere.

[0936] Prepare compound solutions with different concentration gradients. Dissolve DMSO to a concentration of 10mM test compound and 10mM reference compound AZD1775, and dilute the compounds in the culture medium in series, with a total of 9 dose points, and set 2 parallel replicates for each concentration. The cell growth group without compound addition serves as a positive control (maximum signal control), and the culture medium serves as a negative control (minimum signal control), while ensuring that the final DMSO content in each reaction well is 0.2%. After removing the culture medium from the 384-well plate, transfer 25ul of the prepared compounds of different concentrations into the well plate, and incubate the compounds and cells in a cell culture incubator at 37°C, 5% CO2 for 3 days.

[0937] Take the 384-well plate out of the cell culture incubator and let it equilibrate to room temperature for 1 hour. Then add 25ul of Cell Titer-Glo detection reagent to each reaction well. After shaking for 2 minutes, incubate for 10 minutes and read the results (Luminescence) with BMG PHERAStar. Calculate the inhibition rate based on the luminescence signal: first calculate the average value of the positive control (maximum signal control) and the negative control (minimum signal control).

[0938] by To calculate the inhibition rate of different concentrations of compounds on cells. The IC50 of the compound's inhibition of cell activity was calculated by fitting in the log(inhibitor) vs.response–Variableslope mode using GraphPad Prism 6. The fitting equation is: Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*HillSlope)), where Y represents the inhibition rate and X represents the concentration of the known compound after Log.

[0939] According to the above method, the example compounds were subjected to H1299 and MIA PaCa2 in vitro anti-cell proliferation tests. The test results are shown in Table 2, in which the IC 50 The descriptions are classified as follows:

[0940] “-” indicates IC 50 The measured value was greater than 10 μM;

[0941] "+" indicates IC 50 The measured value is less than or equal to 10 μM and greater than 5 μM;

[0942] "++" indicates IC 50 The measured value is less than or equal to 5 μM and greater than 2 μM;

[0943] "+++" indicates IC 50 The measured value is less than or equal to 2 μM and greater than 1 μM;

[0944] "++++" indicates IC 50 The measured value is less than or equal to 1 μM and greater than 0.1 μM;

[0945] "++++++" indicates IC 50 The measured value was less than or equal to 0.1 μM.

[0946] Table 2. In vitro anti-cell proliferation inhibitory activity of the compounds of the present invention on H1299 and MIA PaCa-2

[0947]

[0948]

[0949] As shown in Table 2, most of the compounds of the present invention have good proliferation inhibition activity against H1299 and MIA PaCa-2 tumor cells, and some of the compounds have better proliferation inhibition activity than the control compound AZD1775.

[0950] Experimental Example 3: In vitro liver microsome (mouse and human) test

[0951] 1. Preparation of working fluid:

[0952] The microsomes were taken out of the -80℃ refrigerator, thawed quickly in a 37℃ water bath, and placed on ice for later use. The test sample was diluted with DMSO to prepare a 10mM stock solution, and then diluted with acetonitrile to a 0.5mM secondary stock solution. The microsomes were diluted to 0.75mg / ml using Buffer C; the secondary stock solution was then added to a final concentration of 1.5μM as the working solution. Based on n=2 and 5 time points, 350μL of each compound was prepared and placed on ice before use. NADPH was diluted with Buffer C to a 6mM working solution as the starting solution. An acetonitrile solution containing an internal standard was prepared as a precipitant. Verapamil-HCl was used as the internal standard at a concentration of 4ng / ml.

[0953] 2. Experimental process:

[0954] Take a round-bottom plate, record it as the reaction plate, and dispense the prepared working solution of each compound into the plate according to the number of replicates and time points (the sample at 0h is also added to the reaction plate), 30μL / well; incubate the reaction plate at 37℃ for 10 minutes. Take another pointed-bottom plate, record it as the precipitation plate, and add 135μL of precipitant to each well; after incubation for 10 minutes, transfer the sample at 0h to the precipitation plate, add 15μL of the starting solution, and place the precipitation plate on ice before centrifugation.

[0955] Add enough diluted starting liquid to the dispensing plate to facilitate the pipetting operation.

[0956] The reaction is carried out on a warm incubation shaking device. Use a spray gun to draw 15μL / sample of the starting solution and add it to the reaction plate. Slightly shake and mix to start the reaction. Use a timer to accurately time and record; usually, samples with shorter reaction times are started later, and vice versa, start earlier to leave enough time for multiple plate operations to terminate the reaction. For example: For experiments with four time points of 5, 15, 30, and 60 minutes, the 60-minute one is started first, and the 5-minute one is started later, so as to leave enough operation time for the 5-minute time point group.

[0957] After the reaction time is up, use a spray gun to suck out all the solutions in the reaction plate and add them to the precipitation plate to terminate the reaction at that time point. After all reactions are terminated, shake the precipitation plate on a shaker at 600 rpm for ten minutes to precipitate the protein. Centrifuge at the maximum speed of the centrifuge at 4°C for 15 minutes, take 80 μL of the supernatant, add 320 μL of pure water, mix well, and perform LC-MS analysis.

[0958] 3. Experimental results: as shown in Table 3.

[0959] Table 3 In vitro liver microsome (mouse and human) test results

[0960] Compound <![CDATA[T 1 / 2 / min(M)]]> <![CDATA[T 1 / 2 / min(H)]]> Compound <![CDATA[T 1 / 2 / min(M)]]> <![CDATA[T 1 / 2 / min(H)]]> AZD1775 25.99 38.12 Compound 3 96.25 >120 Compound 4 10.08 22.21 Compound 6 5.72 5.16 Compound 8 >120 Not tested Compound 9 0.78 2.82 Compound 10 1.34 8.31 Compound 11 110.0 Not tested Compound 12 31.50 Not tested Compound 15 >120 Not tested Compound 18 >120 Not tested Compound 19 48.12 Not tested Compound 23 38.72 Not tested Compound 24 >120 Not tested Compound 25 14.09 Not tested Compound 27 33 105 Compound 28 15.03 Not tested Compound 37 21.72 Not tested Compound 39 >120 Not tested Compound 46 13.64 16.82 Compound 51 14.65 19.04 Compound 54 77 Not tested Compound 57 43.86 Not tested Compound 61 >120 Not tested Compound 62 25.11 Not tested / / /

[0961] As shown in Table 3, the compounds of the present invention have good metabolic stability properties under the evaluation of the in vitro liver microsome model.

[0962] Experimental Example 4 Cell Permeability Evaluation (Caco-2 Model)

[0963] Caco-2 cells were purchased from the American Model Tissue Cell Collection (Rockville, MD). The cell culture medium was modified Eagle's medium (MEM) containing 10% inactivated fetal bovine serum and 1% non-essential amino acids. The cells were seeded on polycarbonate filters (Cat. No. 3396) and placed at 37°C, 5% CO 2 Culture in an incubator.

[0964] Cells can be used for transport experiments after 21 to 28 days of culture and the apparent permeability coefficient (Papp ) to characterize and verify the density of the cell monolayer. In the experiment, the compound was dissolved in DMSO to prepare a 10mM stock solution, and diluted with Hanks balanced salt solution (HBSS, Invitrogen, Cat#14025-092) containing 25mM HEPES (pH 7.4) to obtain a working solution. A 10μM working solution of the compound to be tested was added to the apical side and basolateral side of Caco-2 and incubated at 37°C for 90 minutes. After the incubation, the samples on the apical side and basolateral side were diluted, and the concentrations of the compounds on the apical side and basolateral side were detected by LC-MS / MS, and the concentrations of the compounds were quantitatively calculated by the standard curve. The test results are shown in Table 4.

[0965] Table 4 Permeability data of compounds in Caco-2 model

[0966]

[0967] It can be concluded from the experimental results in Table 4 that the cell permeability properties of some compounds of the present invention in the Caco-2 model are comparable to those of the control compounds.

[0968] Experimental Example 5: Evaluation of plasma protein binding rate (PPB)

[0969] 1. Experimental Procedure

[0970] Sample preparation: Dissolve the compound in DMSO to prepare a 10 mM stock solution, then dilute the compound in PBS to prepare a 0.02 mM secondary stock solution, and then use blank plasma to dilute the 0.02 mM solution to 1 μM, which is the sample to be incubated.

[0971] Preparation of dialysis device: first add 400 μL blank PBS to the white well of the balanced dialysis plate, add 200 μL prepared plasma sample to the red well, and seal the dialysis plate with sealing film.

[0972] Recovery plate preparation: Prepare two 96-well deep-well plates, marked as T0 and T5, and add all plasma samples to the two plates with n = 2. Add 300 μL acetonitrile (Verapamil-HCl, 4 ng / mL) directly to the T0 plate, then add 50 μL blank PBS and mix for 5 minutes, and place in a 4°C refrigerator until the end of the incubation experiment.

[0973] Experimental operation: The dialysis device and T5 plate were placed in a microplate constant temperature oscillator and incubated for 5 hours (37°C, 300 rpm or minimum speed). After the incubation, 300 μL of acetonitrile (Verapamil-HCl, 4 ng / mL) was added, and then 50 μL of PBS solution was added. After the dialysis incubation, a new 96-well deep well plate was taken. 50 μL of plasma well sample was added to the corresponding position of the 96-well plate, and then 300 μL of acetonitrile was added, and 50 μL of blank PBS was added; 50 μL of buffer well sample was added to the corresponding position of the 96-well plate, and then 300 μL of acetonitrile was added, and 50 μL of blank plasma was added. 300 μL of acetonitrile (Verapamil-HCl, 4 ng / mL) was added to the plasma well of the T5 plate, and then 50 μL of PBS solution was added. Oscillate for 5 minutes to fully precipitate the protein, and centrifuge at 4°C, 20,000g for 10 minutes. Take 200 μL of supernatant and add it to 200 μL of pure water, mix well and perform LC-MS / MS analysis.

[0974] 2. Data processing and parameter calculation

[0975] ●Plasma protein binding rate = [(Rpe-Rb) / Rpe] × 100%

[0976] ●Recovery rate = [(Rpe+Rb) / R5h] × 100%

[0977] ●Stability = (R5 / R0) × 100%

[0978] in:

[0979] ●R pe = Ratio of the peak area of ​​the test sample on the plasma side to the internal standard

[0980] ●R b = Ratio of peak area of ​​test sample on buffer side to internal standard

[0981] ·R 5 = Ratio of peak area of ​​incubator stability sample to internal standard

[0982] ·R 0 = Ratio of peak area of ​​refrigerator stability sample to internal standard

[0983] 3. The test results are shown in Table 5:

[0984] Table 5 Plasma protein binding rate of representative compounds (mouse)

[0985]

[0986]

[0987] It can be seen from the experimental results in Table 5 that the compounds of the present invention have good plasma protein binding ability.

[0988] Experimental Example 6: Evaluation of the inhibitory activity of CYP450 subtypes

[0989] The enzymatic experiment quantitatively detects the inhibition of the activity of each subtype of CYP450 by small molecule inhibitors through the fluorescence generated by the oxidation of substrates by cytochrome P450. The experiment was carried out in a 384-well plate (Corning, Cat#3575) and the reaction buffer used was: 142.86mM Potassium Phosphate, pH 7.4. The components of Solution A used in the experiment were: 26.13mM NADP + (Sigma-aldrich, Cat#N0505), 65.77mM G6P (J&K, Cat#968161) and 65.42mM MgCl 2 (Sigma-aldrich, Cat#M2670). The components of Solution B used in the experiment were: 40U / mL G6PDH (Sigma-aldrich, Cat#G6378). The components of the substrate mixture solution were: 0.05X Solution A, 0.01X Solution B, 50mM Potassium Phosphate, 0.01mM BOMCC / 0.01mM EOMCC / 0.001mM DBOMF. For CYP3A4 and CYP2C9, the reaction system was 50μL or 20μL, respectively, including 3nM CYP3A4 or 120nM CYP2C9, BOMCC substrate mixture solution and different concentrations of the test compound. For CYP2C19, CYP2D6 and CYP1A2, the reaction system was 20 μL, including 12.5 nM CYP2C19, 80 nM CYP2D6 or 1 nM CYP1A2, EOMCC substrate mixed solution and different concentrations of the test compound. For CYP2C8, the reaction system was 50 μL, including 1.5 nM CYP2C8, DBOMF substrate mixed solution and different concentrations of the test compound. After the compound and enzyme were pre-incubated for 10 minutes, the substrate was added, and the fluorescence signals of different bands (BOMCC / EOMCC Ex430 nm / Em480 nm, DBOMF Ex490 nm / Em520 nm) were read using BMG PHERAStar according to different substrates. The reaction interval was 30 seconds or more (set according to the actual number of experimental wells), and the reaction time was 30 minutes. The experimental data were analyzed and processed by GraphPad Prism 6 software to obtain IC 50 The results are shown in Table 6.

[0990] Table 6 Results of the inhibition test of compounds on different subtypes of CYPs

[0991]

[0992] It can be seen from the experimental results in Table 6 that compound 3 of the present invention has no obvious cytochrome P450 enzyme inhibitory activity.

[0993] Experimental Example 7: Evaluation of potassium channel (hERG) inhibitory activity

[0994] Experimental purpose: To detect the effect of the test compound on hERG potassium channel using manual patch clamp method.

[0995] Experimental steps:

[0996] 1. Experimental materials:

[0997] A. CHO (Chinese Hamster Ovary) Stable Transfection Cell Line Culture

[0998] The cell line used in the patch clamp experiment was the 10th generation CHO cells overexpressing the hERG potassium channel cDNA.

[0999] CHO hERG cells were cultured in a culture dish or culture flask in a 37°C, 5% CO2 incubator. 24-48 hours before the electrophysiological experiment, the cells were dropped onto a round glass slide and cultured in a cell culture medium, and used in the experiment after the cells adhered to the wall.

[1000] Cell culture medium (purchased from Invitrogen) components:

[1001] Ham's F12 medium

[1002] 10% (v / v) heat-inactivated FBS

[1003] 100 μg / ml Hygromycin B

[1004] 100 μg / ml Geneticin (G418)

[1005] B. Compound Preparation

[1006] Compound powders were dissolved in the extracellular fluid, and all were routinely sonicated and shaken for 5 to 10 minutes to ensure complete dissolution of the compound.

[1007] The final concentrations of the compounds used for electrophysiological assays were 5 and 20 μM, and the final concentration of DMSO was 0.1%.

[1008] (II) Experimental plan:

[1009] A. Electrophysiological Recording Experimental Procedure

[1010] Cell membrane currents were recorded using a HEKA EPC-10USB patch clamp amplifier (HEKA Elektronik, Germany).

[1011] 1) Take a coverslip with a large number of single CHO hERG cells growing evenly on the surface, place it in a continuous recording pool on an inverted microscope, perfuse extracellular fluid (about 1 ml per minute) and continue recording until the current stabilizes.

[1012] 2) Use the standard whole-cell recording mode to record the HERG channel current of a single cell. First, clamp the membrane voltage at

[1013] -80mV, give the cells a +20mV voltage stimulus for 5s to activate the hERG potassium channel, then repolarize to -50mV for 5s to generate an outward tail current, and continue perfusing until the current stabilizes. At this time, the peak tail current value is the control current value.

[1014] 3) Then, the extracellular fluid containing the drug to be tested is perfused and the recording is continued until the inhibitory effect of the drug on the hERG current reaches a stable state. At this time, the peak value of the tail current is the current value after drug addition.

[1015] 4) Perfuse the cells again with extracellular solution until the hERG current returns to or approaches the level before drug addition, then continue perfusing to test other concentrations or drugs. One or more compounds or drug concentrations can be tested on each cell.

[1016] 5) Cisapride (C4740-10 mg, Sigma) was used as a positive control in the experiment to ensure that the cells used reacted normally.

[1017] (III) Quality Control

[1018] The test data in the report needs to meet the following standards:

[1019] Electrophysiological recording parameters

[1020] a) Sealing resistance>500MΩ

[1021] b) Contact resistance (Ra) <10MΩ

[1022] c) Initial tail current amplitude > 200pA

[1023] d) Current rundown (spontaneous decrease) <2% / min

[1024] e) Leakage current < 200 pA or 10% of the peak hERG current (within 90% of the recording time)

[1025] The test results are shown in Table 7:

[1026] Table 7 hERG inhibition test results of compounds

[1027]

[1028] It can be seen from the experimental results in Table 7 that compound 3 of the present invention has no obvious hERG inhibitory activity. It can be seen from the experimental results of the present invention that the compounds of the present invention, especially the preferred compounds, are superior to the prior art in terms of drug activity and toxic side effects.

[1029] Experimental Example 8: Pharmacokinetic evaluation of single-dose administration in mice

[1030] This experiment aims to study the pharmacokinetics in plasma of male ICR mice after administration.

[1031] 1. Purpose of the experiment

[1032] The purpose of this study is to obtain the pharmacokinetic profile of the test compound in ICR mice (including intravenous and oral administration). 2. Regulatory Compliance This study (non-GLP study) test sample-testing, DMPK animal testing, and DMPK analysis were all completed in Chengdu Pioneer. All experiments followed this experimental protocol and the relevant SOPs of relevant institutions.

[1033] 3. Test materials, instruments and equipment

[1034] 3.1 Test materials

[1035] 3.1.1 Test products

[1036] The following test samples are provided by Chengdu Pioneer, and their quality is guaranteed to meet the requirements.

[1037] Name / Code Test compound Characteristics Solution Solvent / dosage form 5% DMSO-10% Solutol-85% HPBCD (20%, W / V) Storage conditions Room temperature Test solution concentration 0.2mg / ml (intravenous), 1mg / ml (oral) Test solution volume 3ml, 3ml

[1038] 3.1.2 Test system

[1039] Species: SPF grade male ICR mice

[1040] Weight / age: about 30g

[1041] Quantity: 6

[1042] Source of experimental animals: Weitonglihua

[1043] 4. Experimental process

[1044] 4.1 Solvent preparation

[1045] Intravenous injection solvent: 5% DMSO-10% Solutol-85% HPBCD (20%, W / V)

[1046] “Gastric administration solvent / dosage form: 5% DMSO-10% Solutol-85% HPBCD (20%, W / V)”

[1047] 4.2 Administration of Test Article (Route)

[1048] Intravenous injection: Dosage 1 mg / kg, administration volume 5 mL / kg

[1049] Oral gavage: dosage 10 mg / kg, administration volume 10 mL / kg

[1050] Ultrasound 5 minutes before administration

[1051] 4.3 Test substance preparation (concentration)

[1052] Intravenous: 0.2 mg / mL

[1053] Intragastric administration: 1 mg / mL

[1054] The subjects were fasted overnight before administration and fed four hours after administration.

[1055] 4.4 Sample collection

[1056] Blood (40-50 μL) was collected by orbital venous puncture at 5 minutes (IV only), 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, and 24 hours after administration into anticoagulant tubes pre-sprayed with EDTA-K2. The blood samples were centrifuged at 10,000 rpm for 20 minutes within 1 hour (stored on wet ice before and after centrifugation), and the upper layer, i.e. plasma, was collected and frozen at -20°C or below for LC-MS / MS analysis.

[1057] 4.5 Analysis and detection of biological drugs: LC-MS / MS analysis and detection.

[1058] 5 Data processing

[1059] The drug concentration of the samples at each time point was detected by LC-MS / MS. Phoenix WinNonlin 5.2 non-compartmental model was used to calculate the pharmacokinetic parameters of terminal elimination half-life (t 1 / 2 ), area under the concentration-time curve (AUC), apparent distribution volume (Vd), clearance (CL), mean residence time (MRT), Cmax, and bioavailability (F%) are directly obtained from serum concentration results. Blood drug concentration and pharmacokinetic parameters are expressed as mean ± standard deviation (X ± SD). Specific detection and analysis methods are specified in the form of protocol revisions.

[1060] 6 The test results are shown in Table 8:

[1061] Table 8 Pharmacokinetic parameters in mice

[1062]

[1063]

[1064] As shown in Experimental Table 8, the pharmacokinetic analysis in mice revealed that the pharmacokinetic properties of compounds 48 and 49 of the present invention were comparable to those of the control compound AZD1775, and the pharmacokinetic properties of compounds 51 and 53 were superior to those of the control compound AZD1775.

Claims

1. A compound represented by formula I or a pharmaceutically acceptable salt thereof: in, The R1 is selected from methyl, ethyl, propyl, isopropyl, cyclopropyl, 3-10 membered heterocyclic ring, the 3-10 membered heterocyclic ring may be further replaced by one, two, three or four independent R 11 Substitute; said R 11 Selected from hydrogen, C 1～6 Alkyl, halogen substituted C 1～6 Alkyl, halogen; R2 is selected from The R 21 Selected from hydrogen, C 1～6 Alkyl, -C 0～2 Alkylene-C(O)OC 1～6 Alkyl, -C 0～2 Alkylene -COOH, -C(O)C 1～6 Alkyl, halogen; The R3 is selected from X1 is selected from chemical bonds, O, NR N1 or CR C1 R C2 ; X2 is selected from N or CR C1 ; The X3 is selected from O, NR N1 or CR C1 R C2 ; n is selected from 1, 2 or 3; R C1 , R C2 are independently selected from hydrogen, C 1～3 Alkyl, halogen, halogen-substituted C 1～3 Alkyl, -C 1～2 Alkylene-N(C 1～3 Alkyl)(C 1～3 alkyl); R N1 Selected from hydrogen, C 1～3 Alkyl, halogen substituted C 1～3 Alkyl, -(3- to 10-membered heterocycloalkyl); The R4 is selected from: Wherein, X is selected from O, NR 41 ; X4 is selected from O, NH or CH2; n1 is selected from 1 or 2; said R4 can be further replaced by one, two, three or four independent R 41 replace; The R 41 Selected from hydrogen, oxo, C 1～3 Alkyl, -C 2～6 Alkenyl, halogen-substituted C 1～3 Alkyl, hydroxy substituted C 1～6 Alkyl, -C 0～2 Alkylene-O(C 1～3 Alkyl), -NH2, -NH(C 1～3 Alkyl), -N(C 1～3 Alkyl)(C 1～3 Alkyl), -C(O)R 42 、-C(O)NR 42 R 43 、-C(O)OR 42 , -(3- to 10-membered cycloalkyl), -(3- to 10-membered heterocycloalkyl), -C 1～2 Alkylene-(5-10 membered aromatic ring); wherein the alkyl, alkylene, cycloalkyl, heterocycloalkyl, aromatic ring may be further replaced by one, two, three or four independent R 44 replace; R 42 , R 43 are independently selected from hydrogen, halogen, -OH, C 1～3 Alkyl, halogen substituted C 1～3 alkyl; R 44 Selected from hydrogen, C 1～3 Alkyl, halogen substituted C 1～3 Alkyl, -O(C 1～3 alkyl); Alternatively, the R 41 With R C1 or R C2 Together with the atoms to which it is directly attached, it forms a 3- to 10-membered heterocyclic ring; The R5 is selected from hydrogen, halogen, -C 1～3 Alkyl, halogen substituted C 1～3 Alkyl, hydroxy substituted C 1～3 Alkyl, -O(C 1～3 Alkyl), -NH(C 1～3 Alkyl), -O-C2 alkylene-N(C 1～3 Alkyl)(C 1～3 alkyl), -O-(3- to 10-membered cycloalkyl), -O-(3- to 10-membered heterocycloalkyl), wherein the alkyl, alkylene, cycloalkyl, heterocycloalkyl may be further replaced by one, two, three or four independent R 51 replace; The R 51 Selected from hydrogen, C 1～3 Alkyl, halogen substituted C 1～3 alkyl; Wherein, the A ring is selected from Wherein, the B ring is selected from R7, R8 and the atoms they are directly connected to form Wherein, the C ring is selected from The R 10 Selected from hydrogen, C 1～6 Alkyl, halogen substituted C 1～6 alkyl; The R9 is selected from -C 0～2 Alkylene-C(O)NR 91 R 92 、-C(O)C 1～6 Alkyl; each R 91 , R 92 are independently selected from hydrogen, C 1～6 Alkyl, halogen substituted C 1～6 alkyl; or R 91 , R 92 The atoms directly connected to it form a 3-10 membered heterocyclic ring, which may be further separated by one, two, three or four independent R 93 replace; Each R 93 are independently selected from hydrogen, C 1～6 Alkyl, halogen substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 Alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl); The 3- to 10-membered heterocyclic ring and the 3- to 10-membered heterocycloalkyl group are saturated rings or non-aromatic unsaturated rings having at least one heteroatom, and the heteroatom is a nitrogen atom, an oxygen atom or a sulfur atom.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The R1 is selected from methyl, ethyl, propyl, isopropyl, cyclopropyl, And / or, said R2 is selected from 3. The compound according to claim 2 or a pharmaceutically acceptable salt thereof, characterized in that: The R4 is specifically selected from:

4. The compound according to claim 3 or a pharmaceutically acceptable salt thereof, characterized in that: The R5 is specifically selected from methyl, ethyl, ethoxy, F, hydroxy-substituted methyl, 5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The R3 is selected from 6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The compound described in formula I is specifically:

7. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The compound described in formula I is specifically:

8. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The compound described in formula I is specifically:

9. Use of the compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a WEE1-mediated disease.

10. The use according to claim 9, characterized in that: The WEE1-mediated disease is one or more of diseases associated with inflammation, autoimmune diseases, infectious diseases, cancer, and precancerous syndrome.

11. A pharmaceutical composition, characterized in that: The invention relates to a preparation prepared by using the compound described in any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof as a pharmaceutically active ingredient and adding pharmaceutically acceptable auxiliary materials.

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