Camptothecin derivative, linker, ligand-drug conjugate and medical application of camptothecin derivative, linker and ligand-drug conjugate

Abstract

The invention discloses a novel camptothecin derivative, a joint, a ligand-drug conjugate, a pharmaceutical composition containing the camptothecin derivative or conjugate, and corresponding medical application.

Description

Camptothecin derivatives, linkers, ligand-drug conjugates and their medical uses Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to camptothecin derivatives, linkers, ligand-drug conjugates and their medical uses in treating or preventing proliferative diseases. Background Art

[0002] Camptothecin is a water-insoluble cytotoxic alkaloid produced from the tree Camptotheca accuminata grown in China and the tree Nothapodytes foetida grown in India. Camptothecin and its related analogs are known potential anticancer agents and have demonstrated therapeutic activity in vitro and in vivo.

[0003] Camptothecin and its analogs are known inhibitors of DNA topoisomerase I. For example, exitecan, an analog of camptothecin, is also a DNA topoisomerase I inhibitor and exhibits strong anticancer activity.

[0004] Ixitectcan, a small molecule anti-tumor compound, is a camptothecin derivative known to exhibit anti-tumor effects by inhibiting DNA topoisomerase I. Developed by Daiichi Sankyo, it was initially advanced to Phase III clinical trials as a standalone chemotherapy agent, with primary indications for bone cancer, prostate cancer, breast cancer, and pancreatic cancer. Unlike irinotecan, currently in clinical use, Ixitectcan does not require enzyme activation. Furthermore, compared to SN-38, the active ingredient in irinotecan, and topotecan, also in clinical use, Ixitectcan exhibits stronger topoisomerase I inhibitory activity and exhibits enhanced cytotoxicity against various cancer cells in vitro. Ixitectcan has not yet been successfully marketed as a standalone chemotherapy agent, presumably due to its high cellular activity, resulting in a narrow therapeutic window.

[0005] Ligand-drug conjugates (ADCs), as a new type of targeted therapy, generally consist of three components: an antibody or antibody-like ligand, a small molecule drug, and a linker that connects the ligand and drug. Antibody-drug conjugates leverage the antibody's specific recognition of the antigen to deliver the drug molecule to the vicinity of the target cell and effectively release the drug molecule, achieving therapeutic efficacy. In 2000, Pfizer's ADC drug Mylotarg debuted on the market, bringing the ADC field, a field brimming with both potential and challenges, into the public consciousness. In recent years, the pharmaceutical market has experienced a new wave of ADC research and development, with 13 ADCs currently on the market worldwide.

[0006] DS-8201a (trade name: Enhertu), an antibody-drug conjugate jointly developed and commercialized by Daiichi Sankyo and AstraZeneca, was launched in December 2019. This drug forms an amide derivative of exotecan and glycolic acid and connects them to form an ADC. As a new generation of antibody-drug conjugate, Enhertu has shown the potential to become a blockbuster.

[0007] It is extremely necessary and urgent to explore and discover camptothecin or exotecan derivatives with better anti-tumor activity, improve the safety and efficacy of small molecule anti-tumor compounds in ADC drug applications, and thus obtain anti-tumor drugs with excellent therapeutic effects.

[0008] Summary of the Invention

[0009] The first aspect of the present invention provides a compound, such as a compound shown in Formula 1 or a pharmaceutically acceptable salt thereof,

[0010] in,

[0011] R 16 、R 17 、R 19 Each occurrence is independently selected from hydrogen, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl; preferably, R 16 、R 17 、R 19 Each occurrence is independently selected from hydrogen and C optionally substituted by R 1-6 Alkyl; More preferably, R 16 、R 17 、R 19 Each occurrence is independently selected from hydrogen and C 1-6 Alkyl; further preferably, R 16 、R 17 、R 19 Each occurrence of is independently selected from hydrogen;

[0012] R 20 Selected from N, and CR 20a;

[0013] R 20a selected from hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -C(O)CH2OH, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, and C 1-6 Alkyl; preferably, R 20a is hydrogen or C 1-6 alkyl;

[0014] R 11 Selected from O and S;

[0015] Q is selected from 0, 1, 2, 3, and 4;

[0016] R 18 selected from H, and -B-R3;

[0017] When Q is 0, R 18 When it is -B-R3, R 14 and R 15 are independently selected from hydrogen atoms, halogens, C 1-8 Alkyl, and C 1-8 preferably, R 14 and R 15 Each independently selected from hydrogen atom, F, Cl, Br, I, C 1-3 Alkyl and C 1-3 preferably, R 14 and R 15 Each independently is -CH3 or F; preferably, R 14 is -CH3, and / or R 15 is -F;

[0018] B is selected from -C(O)- and -P(O)(OH)-; preferably B is -C(O)-;

[0019] R3 is selected from -(CH2CH2O) a C 1-6 alkyl, Among them, when -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m-, the n methylene units are each independently replaced by -N(R5)C(O)-, -C(O)-, -OC(O)-, -NR5-, -O-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -C(=S)-, -C(=NR5)-, -N=N- or -N=CH-;

[0020] a is an integer greater than or equal to 1, preferably, a is an integer from 1 to 20, and further preferably, a is selected from 1, 2, 3, 4, 5, 6, 7, and 8;

[0021] R4 and R8 are each independently a single bond or are independently selected from -O-, C 1-8 Alkylene, C 1-8 Haloalkylene, -N(R5)C(O)-, -C(O)-, -OC(O)-, -NR5-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -C(=S)-, -C(=NR5)-, -N=N- and -N=CR5-;

[0022] Ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from 6-10 membered aryl or arylene, 5-10 membered heteroaryl or heteroarylene, 3-10 membered cycloalkyl or cycloalkylene, and 3-10 membered heterocyclyl or heterocyclylene, preferably, the heteroaryl, heteroarylene, heterocyclylene and heterocyclyl each independently contain 1, 2, 3 or 4 heteroatoms independently selected from N, O, P and S; and when R3 is When W1 and W2 are fused together, when R3 is

[0023] When , W1 and W2 are fused together, W2 and W3 are fused together, and W3 and W4 are fused together;

[0024] When ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from 3-10 membered cycloalkyl, cycloalkylene, heterocyclylene, and heterocyclyl, the 3-10 membered cycloalkyl, cycloalkylene, heterocyclylene, and heterocyclyl are optionally replaced by one or more R w1 、R w2 、R w3 、R w4 , or R w5 Replacement, R w1 、R w2 、R w3 、 R w4 , and R w5Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halogen, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Alkyl; preferably, R w1 、R w2 、R w3 、R w4 , and R w5 Each occurrence is independently selected from hydrogen, deuterium, F, Cl, Br, I, hydroxyl, ═O, ═S, —NH 2 , —NO 2 , —CN, —COOH, —OC 1-3 Alkyl, -C 1-3 Alkyl, and C 1-3 alkylene hydroxyl group;

[0025] When ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from 6-10 membered aryl, 6-10 membered arylene, 5-10 membered heteroarylene, and 5-10 membered heteroaryl, the aryl, arylene, heteroarylene, and heteroaryl groups are optionally replaced by one or more R w1 、R w2 、R w3 、R w4 , or R w5 Replacement, R w1 、R w2 、R w3 、R w4 , and R w5 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Alkyl; preferably, R w1 、R w2 、Rw3 、R w4 , and R w5 Each occurrence is independently selected from hydrogen, deuterium, F, Cl, Br, I, hydroxyl, -NH2, -NO2, -CN, -OC 1-3 Alkyl, -COOH, and C 1-3 alkyl;

[0026] R 13 Selected from hydrogen atoms, halogens, C 1-8 Alkyl, and C 1-8 preferably, R 13 is a hydrogen atom; R 12 Selected from C 1-8 Alkylene N(R a )(R b ), C 1-8 Haloalkylene N(R a )(R b ), halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Alkyl; preferably, R 12 Selected from C 1-3 Alkylene N(R a )(R b ), and C 1-3 Haloalkylene N(R a )(R b ); preferably, R 12 Selected from -CH2N(R a )(R b ); or, R 13 and R 12 Together with the carbon atom to which it is attached, it forms a saturated or unsaturated 5-10 membered cycloalkyl or a 5-10 membered heterocyclic group containing 1, 2 or 3 heteroatoms independently selected from N, O, P and S, wherein the cycloalkyl or heterocyclic group is optionally substituted by one or more independently selected from halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(Ra )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 The substituent of the alkyl group is substituted; preferably, R 13 and R 12 Together with the carbon atom to which it is attached, a 6-membered cycloalkyl group is formed, wherein the cycloalkyl group is -N(R a )(R b )replace;

[0027] R 3a , R 3b , and R5 are each independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 alkyl; and

[0028] m, and n are each independently an integer greater than or equal to 1, and m≥n; preferably, m and n are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m≥n; preferably, m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, -n is 0, 1, 2, 3, 4, or 5, and m≥n;

[0029] When Q is 1, 2, 3 or 4, R 18 When H, R 12 、R 13 , and R 14 are each independently selected from hydrogen, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Alkyl; preferably, R12 、R 13 , and R 14 Each independently selected from hydrogen, F, Cl, Br, I, C 1-3 Alkyl, -NO2, -CN, -OC 1-3 Alkyl; or R 12 and R 13 Together with the carbon atom to which it is attached, it forms a saturated or unsaturated 5-10 membered cycloalkyl or a 5-10 membered heterocyclic group containing 1, 2 or 3 heteroatoms independently selected from N, O, and S, wherein the cycloalkyl or heterocyclic group is unsubstituted or substituted by one or more independently selected from halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 The substituent of the alkyl group is substituted; preferably, R 12 and R 13 Together with the carbon atom to which it is attached, it forms a saturated or unsaturated 5-6 membered heterocyclic group containing 2 O atoms;

[0030] R 15 C 1-8 Alkylene hydroxyl, C 1-8 Alkyleneamino, -C(R a )(R b )-N(R a )-R 4c 、-C(R a )(R b )-N(R a )-C(O)-R 4c , or -C(R a )(R b )-N(R a )-C(O)OR 4c Preferably, R 15 -CH2-NH2, -CH2-NH-C(O)-R 4c 、-CH2-NH-C(O)OR 4c , or C4H8OH; preferably, R 15 is -CH2-NH2, or -(CH2)4-OH; and

[0031] R, R a, R b , and R 4c Each occurrence is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -C(O)CH2OH, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 Alkyl or C 1-6 Alkylene hydroxyl; preferably, R, R a , R b ,R 4c Each occurrence is independently hydrogen, F, Cl, Br, I, CH2OH, or C2H4OH;

[0032] When Q is 0, R 11 S, R 18 When H, R 12 selected from hydrogen, and -CH2N(R 3A )(R 4A );

[0033] R 14 , and R 15 are each independently selected from hydrogen, halogen, -NO2, -CN, -OR', -N(R a1 )(R b1 ), -C(O)R', -CO2R', -C(O)C(O)R', -C(O)CH2C(O)R', -C(O)N(R a1 )(R b1 )、-SO2N(R a1 )(R b1 ), -OC(O)R', -N(R')SO2R', and C 1-6 Alkyl; preferably, R 14 and R 15 are independently selected from halogen, amino, -OR' and C 1-6 Alkyl; further preferably, R 14 and R 15 Each independently selected from fluorine, chlorine, amino, -OC 1-6 Alkyl and C 1-6 alkyl;

[0034] or R 14 and R 15can form together a saturated or unsaturated 4-10 membered cycloalkyl or a 5-membered or 6-membered heterocyclic group containing 1-3 heteroatoms independently selected from N, O, and S, wherein the cycloalkyl or heterocyclic group is unsubstituted or substituted by one or more independently selected halogens, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl substitution; preferably, the cycloalkyl or heterocyclyl is unsubstituted;

[0035] R 13 For hydrogen, -C 1-6 Alkyl or -C 1-6 Alkylene-hydroxy; preferably, R 5A For hydrogen, C 1-6 Alkyl or -(CH2)4-hydroxy;

[0036] R 3A and R 4A Each occurrence is independently selected from hydrogen, halogen, -OR', -N(R a1 )(R b1 ), -C(O)R', -C(S)R', -C(S)C(R a1 )(R b1 )OH、-C(O)CH2N(R a1 )(R b1 ), -CO2R', -C(O)C(O)R', -C(O)CH2C(O)R', -C(O)C(R a1 )(R b1 )OH, -C(O)-C 3-6 Cycloalkylene-OH, -C(O)-C 1-6 Alkylene-OH, -C 1-6 Alkylene-OH-C(O)-C 3-6 Cycloalkylene-C 1-6 Alkylene -OH, -C(O)OC 1-6 Alkylene-OH, -S(O)R', -S(O)2R, -C(O)N(R a1 )(R b1 )、-SO2N(R a1)(R b1 ), -OC(O)R', -N(R')SO2R', and C optionally substituted by R' 1-6 Alkyl, C 1-6 Alkenyl and C 1-6 Alkynyl; preferably, R 4A Selected from hydrogen, or C 1-6 alkyl;

[0037] where R', R a1 , and R b1 Each occurrence is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl, 3, 4, 5, 6, 7 or 8 membered cycloalkyl, 3-8 membered heterocyclyl containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, 6-10 membered aryl or 5-10 membered heteroaryl containing 1, 2, or 3 heteroatoms independently selected from N, O, and S; said C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 The alkynyl, 3-, 4-, 5-, 6-, 7-, or 8-membered cycloalkyl, 3- to 8-membered heterocyclyl, 6- to 10-membered aryl, or 5- to 10-membered heteroaryl groups are unsubstituted or substituted with one or more substituted radicals independently selected from halogen, -NO2, -CN, -OH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 Alkyl, C 2-6 Alkenyl or C 1-6 Alkynyl, and hydroxyl C 1-6 Alkylene substitution;

[0038] When R 14 -CH3, R 15 is fluorine, R 13 When it is hydrogen, R 12 for and

[0039] Among them, when R 14 for -O-CH3, R 15 is fluorine, R 13 When it is hydrogen, R12 Not for and

[0040] Among them, when R 14 With R 15 Forming 1,4-dioxo six-membered ring, R 13 When it is hydrogen, R 12 -CH2NH2, -CH2NHC2H4OH, -CH2N(CH3)C2H4OH, -CH2NHC3H6OH, and

[0041] Among them, when R 14 is fluorine, R 15 is fluorine, R 13 When it is hydrogen, R 12 is not hydrogen; and

[0042] Among them, when R 14 -CH3, R 15 is chlorine, R 13 When it is hydrogen, R 12 Selected from and

[0043] Among them, when R 14 With R 15 Forming a 1,3-dioxo five-membered ring, R 13 When it is hydrogen, R 12 Selected from -CH2NHC2H4OH, -CH2N(CH3)C2H4OH, -CH2NHC3H6OH, or

[0044] Among them, when R 13 When selected from -(CH2)4-OH, R 12 When it is hydrogen, R 14 、R 15 is fluorine, or R 14 With R 15 Forming a 5-membered or 6-membered heterocyclic ring containing two oxygen atoms.

[0045] In some embodiments, the structure of Formula 1 is selected from the following structures:

[0046] When Formula 1 is Formula 1A:

[0047] Wherein, R1 and R2 are each independently selected from hydrogen atom, halogen, C 1-8 Alkyl, and C 1-8Preferably, R1 and R2 are each independently selected from hydrogen atom, F, Cl, Br, I, C 1-3 Alkyl and C 1-3 Haloalkyl; preferably, R1 and R2 are each independently -CH3 or F; preferably, R1 is -CH3, and / or R2 is F;

[0048] A is selected from oxygen atoms and sulfur atoms;

[0049] B is selected from -C(O)- and -P(O)(OH)-; preferably B is -C(O)-;

[0050] R3 is selected from -(CH2CH2O) a C 1-6 alkyl, Among them, when -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m -, the n methylene units are each independently replaced by -N(R5)C(O)-, -C(O)-, -OC(O)-, -NR5-, -O-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -C(=S)-, -C(=NR5)-, -N=N- or -N=CH-;

[0051] a is an integer greater than or equal to 1, preferably, a is an integer from 1 to 20, and further preferably, a is selected from 1, 2, 3, 4, 5, 6, 7, and 8;

[0052] R4 and R8 are each independently a single bond or are independently selected from -O-, C 1-8 Alkylene, C 1-8 Haloalkylene, -N(R5)C(O)-, -C(O)-, -OC(O)-, -NR5-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -C(=S)-, -C(=NR5)-, -N=N- and -N=CR5-;

[0053] Ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from 6-10 membered aryl or arylene, 5-10 membered heteroaryl or heteroarylene, 3-10 membered cycloalkyl or cycloalkylene, and 3-10 membered heterocyclyl or heterocyclylene, preferably, the heteroaryl, heteroarylene, heterocyclylene and heterocyclyl each independently contain 1, 2, 3 or 4 heteroatoms independently selected from N, O, P and S; and when R3 is When W1 and W2 are fused together, when R3 is

[0054] When , W1 and W2 are fused together, W2 and W3 are fused together, and W3 and W4 are fused together;

[0055] When ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from 3-10 membered cycloalkyl, cycloalkylene, heterocyclylene, and heterocyclyl, the 3-10 membered cycloalkyl, cycloalkylene, heterocyclylene, and heterocyclyl are optionally replaced by one or more R w1 、R w2 、R w3 、R w4 , or R w5 Replacement, R w1 、R w2 、R w3 、R w4 , and R w5 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halogen, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Alkyl; preferably, R w1 、R w2 、R w3 、R w4 , and R w5 Each occurrence is independently selected from hydrogen, deuterium, F, Cl, Br, I, hydroxyl, ═O, ═S, —NH 2 , —NO 2 , —CN, —COOH, —OC 1-3 Alkyl, -C 1-3 Alkyl, and C 1-3 alkylene hydroxyl group;

[0056] When ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from 6-10 membered aryl, 6-10 membered arylene, 5-10 membered heteroarylene, and 5-10 membered heteroaryl, the aryl, arylene, heteroarylene, and heteroaryl groups are optionally replaced by one or more R w1 、R w2 、R w3 、R w4 , or R w5 Replacement, R w1 、R w2 、R w3 、R w4 , and R w5 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Alkyl; preferably, R w1 、R w2 、R w3 、R w4 , and R w5 Each occurrence is independently selected from hydrogen, deuterium, F, Cl, Br, I, hydroxyl, -NH2, -NO2, -CN, -OC 1-3 Alkyl, -COOH, and C 1-3 alkyl;

[0057] R6 is selected from hydrogen atom, halogen, C 1-8 Alkyl, and C 1-8 Preferably, R6 is a hydrogen atom; R7 is selected from C 1-8 Alkylene N(R a )(R b ), C 1-8 Haloalkylene N(R a )(R b ), halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b)、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, -O-CH2C(O)N(R a )(R b ) and C optionally substituted by R 1-6 Alkyl; preferably, R7 is selected from C 1-3 Alkylene N(R a )(R b ), and C 1-3 Haloalkylene N(R a )(R b ); Preferably, R7 is selected from -CH2N(R a )(R b ); or, R6 and R7 together with the carbon atom to which they are attached form a saturated or unsaturated 5-10 membered cycloalkyl or a 5-10 membered heterocyclic group containing 1, 2 or 3 heteroatoms independently selected from N, O, P and S, wherein the cycloalkyl or heterocyclic group is optionally substituted by one or more independently selected halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Preferably, R6 and R7 form a 6-membered cycloalkyl group with the carbon atom to which they are attached, and the cycloalkyl group is replaced by -N(R a )(R b )replace;

[0058] R 3a , R 3b , and R5 are each independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 alkyl;

[0059] R, R a , and R b Each occurrence is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -C(O)CH2OH, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 Alkylene hydroxyl or C 1-6 alkyl; and

[0060] m, and n are each independently an integer greater than or equal to 1, and m≥n; preferably, m and n are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m≥n; preferably, m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, -n is 0, 1, 2, 3, 4, or 5, and m≥n;

[0061] When Formula 1 is Formula 1B:

[0062] Where q is 1, 2 or 3;

[0063] R 1c 、R 2c , and R 5c are each independently selected from hydrogen, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Alkyl; preferably, R 1c 、R 2c , and R 5c Each independently selected from hydrogen, F, Cl, Br, I, C 1-3 Alkyl, -NO2, -CN, -OC 1-3 Alkyl; or R 1c and R 2c Together with the carbon atom to which it is attached, it forms a saturated or unsaturated 5-10 membered cycloalkyl or a 5-10 membered heterocyclic group containing 1, 2 or 3 heteroatoms independently selected from N, O, and S, wherein the cycloalkyl or heterocyclic group is unsubstituted or substituted by one or more independently selected from halogen, -NO2, -CN, -OR, -SR, -N(Ra )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 The substituent of the alkyl group is substituted; preferably, R 1c and R 2c Together with the carbon atom to which it is attached, it forms a saturated or unsaturated 5-6 membered heterocyclic group containing 2 O atoms;

[0064] R 6c C 1-8 Alkylene hydroxyl, C 1-8 Alkyleneamino, -C(R a )(R b )-N(R a )-R 4c 、-C(R a )(R b )-N(R a )-C(O)-R 4c , or -C(R a )(R b )-N(R a )-C(O)OR 4c Preferably, R 6c -CH2-NH2, -CH2-NH-C(O)-R 4c 、-CH2-NH-C(O)OR 4c , or C4H8OH; preferably, R 6c is -CH2-NH2, or -(CH2)4-OH; and

[0065] R, R a , R b , and R 4c Each occurrence is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -C(O)CH2OH, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 Alkyl or C 1-6 Alkylene hydroxyl; preferably, R, R a , R b ,R4c Each occurrence is independently hydrogen, F, Cl, Br, I, CH2OH, or C2H4OH;

[0066] When Formula 1 is Formula 1C:

[0067] R 6A selected from hydrogen, and -CH2N(R 3A )(R 4A );

[0068] R 1A , and R 2A are each independently selected from hydrogen, halogen, -NO2, -CN, -OR', -N(R a1 )(R b1 ), -C(O)R', -CO2R', -C(O)C(O)R', -C(O)CH2C(O)R', -C(O)N(R a1 )(R b1 )、-SO2N(R a1 )(R b1 ), -OC(O)R', -N(R')SO2R', and C 1-6 Alkyl; preferably, R 1A and R 2A are independently selected from halogen, amino, -OR' and C 1-6 Alkyl; further preferably, R 1A and R 2A Each independently selected from fluorine, chlorine, amino, -OC 1-6 Alkyl and C 1-6 alkyl;

[0069] or R 1A and R 2A can form together a saturated or unsaturated 4-10 membered cycloalkyl or a 6-membered heterocyclic group containing 1-3 heteroatoms independently selected from N, O, and S, wherein the cycloalkyl or heterocyclic group is unsubstituted or substituted by one or more independently selected from halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C1-6 alkyl, C1-6 alkenyl or C1-6 alkynyl optionally substituted by R; preferably, the cycloalkyl or heterocyclyl is unsubstituted;

[0070] R 5A is hydrogen or C 1-6 alkyl;

[0071] R 3A and R 4A Each occurrence is independently selected from hydrogen, halogen, -OR', -N(R a1 )(R b1 ), -C(O)R', -C(S)R', -C(S)C(R a1 )(R b1 )OH、-C(O)CH2N(R a1 )(R b1 ), -CO2R', -C(O)C(O)R', -C(O)CH2C(O)R', -C(O)C(R a1 )(R b1 )OH, -C(O)-C 3-6 Cycloalkylene-OH, -C(O)-C 1-6 Alkylene-OH, -C(O)-C 3-6 Cycloalkylene-C 1-6 Alkylene -OH, -C(O)OC 1-6 Alkylene-OH, -S(O)R', -S(O)2R, -C(O)N(R a1 )(R b1 )、-SO2N(R a1 )(R b1 ), -OC(O)R', -N(R')SO2R', and C optionally substituted by R' 1-6 Alkyl, C 1-6 Alkenyl and C 1-6 Alkynyl; preferably, R 4A Selected from hydrogen, or C 1-6 alkyl;

[0072] where R', R a1 , and R b1 Each occurrence is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6Alkynyl, 3, 4, 5, 6, 7 or 8-membered cycloalkyl, 3-8 membered heterocyclyl containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, 6-10 membered aryl or 5-10 membered heteroaryl containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, wherein the C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 The alkynyl, 3-, 4-, 5-, 6-, 7-, or 8-membered cycloalkyl, 3- to 8-membered heterocyclyl, 6- to 10-membered aryl, or 5- to 10-membered heteroaryl groups are unsubstituted or substituted with one or more substituted radicals independently selected from halogen, -NO2, -CN, -OH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 Alkyl, C 2-6 Alkenyl or C 1-6 Alkynyl, and hydroxyl C 1-6 Alkylene substitution;

[0073] When R 1A -CH3, R 2A is fluorine, R 5A When it is hydrogen, R 6A for and

[0074] When R 1A for -O-CH3, R 2A is fluorine, R 5A When it is hydrogen, R 6A Not for and

[0075] When R 1A With R 2A Forming 1,4-dioxo six-membered ring, R 5A When it is hydrogen, R 6A -CH2NH2, -CH2NHC2H4OH, -CH2N(CH3)C2H4OH, -CH2NHC3H6OH, and

[0076] When R 1A is fluorine, R 2A is fluorine, R 5A When it is hydrogen, R 6A is not hydrogen; and

[0077] When R1A -CH3, R 2A is chlorine, R 5A When it is hydrogen, R 6A Selected from and

[0078] When R 1A With R 2A Forming a 1,3-dioxo five-membered ring, R 5A When it is hydrogen, R 6A Selected from -CH2NHC2H4OH, -CH2N(CH3)C2H4OH, -CH2NHC3H6OH, or

[0079] Among them, when R 5A When selected from -C4H8OH, R 6A is hydrogen, R 1A 、R 2A is fluorine, or R 1A With R 2A Forming a 5-membered or 6-membered heterocyclic ring containing two oxygen atoms.

[0080] In some embodiments, each occurrence of Ring W1, Ring W2, Ring W3, Ring W4, and Ring W5 is independently selected from the group consisting of 6- or 8-membered aryl or arylene, 5-, 6-, 7-, or 8-membered heteroaryl or heteroarylene containing 1, 2, 3, or 4 heteroatoms independently selected from N, O, and S, 4-, 5-, 6-, 7-, or 8-membered cycloalkyl or cycloalkylene, and 4-, 5-, 6-, 7-, or 8-membered heterocyclyl or heterocyclylene containing 1, 2, 3, or 4 heteroatoms independently selected from N, O, and S. When Ring W1, Ring W2, Ring W3, Ring W4, and Ring W5 are each independently selected from the group consisting of 4-8-membered cycloalkyl, cycloalkylene, heterocyclylene, and heterocyclyl, the 4-, 5-, 6-, 7-, or 8-membered cycloalkyl, cycloalkylene, heterocyclylene, and heterocyclyl are optionally substituted with one or more R w1 、R w2 、R w3 、R w4 , or R w5 Replacement, R w1 、R w2 、R w3 、R w4 , and R w5 Each occurrence is independently selected from hydrogen, deuterium, F, Cl, Br, I, hydroxyl, ═O, ═S, —NH 2 , —NO 2 , —CN, —COOH, —OC 1-3 Alkyl, -C 1-3 Alkyl, and C 1-3Alkylene hydroxyl; when ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from 6 or 8-membered aryl, 6 or 8-membered arylene, 5, 6, 7 or 8-membered heteroarylene and heteroaryl, the aryl, arylene, heteroarylene and heteroaryl are optionally replaced by one or more R w1 、R w2 、R w3 、R w4 , or R w5 Replacement, R w1 、R w2 、R w3 、R w4 , and R w5 Each occurrence is independently selected from hydrogen, deuterium, F, Cl, Br, I, hydroxyl, -NH2, -NO2, -CN, -OC 1-3 Alkyl, -COOH, and C 1-3 Preferably, ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently optionally substituted with hydrogen, deuterium, F, Cl, Br, I, hydroxyl, -NH2, -NO2, -CN, -OC 1-3 Alkyl, -COOH, or C 1-3 Alkyl-substituted phenyl or phenylene, pyridyl or pyridylene, pyrazolyl or pyrazolylene, or or is optionally substituted with hydrogen, deuterium, F, Cl, Br, I, hydroxyl, ═O, ═S, —NH 2 , —NO 2 , —CN, —COOH, —OC 1-3 Alkyl, -C 1-3 Alkyl, or C 1-3 Alkylene hydroxy-substituted tetrahydropyranyl or tetrahydropyranylene, dioxolanyl or dioxolanylene, tetrahydrofuranyl or tetrahydrofuranylene, represents the site of attachment; and / or

[0081] When R3 is When , ring W1 and ring W2 are fused to form the following fused ring structure:

[0082] Among them, R w1 and R w2 Each occurrence is independently selected from hydrogen, deuterium, F, Cl, Br, I, hydroxyl, ═O, ═S, —NH 2 , —NO 2 , —CN, —COOH, —OC 1-3 Alkyl, -C 1-3 Alkyl, and C 1-3 Alkylene hydroxyl group, represents the site of attachment; and / or

[0083] R 3AEach occurrence is independently selected from hydrogen, -C(O)R', -C(S)R', -C(S)C(R a1 )(R b1 )OH、-C(O)CH2N(R a1 )(R b1 )、-CO2R'、-C(O)C(R a1 )(R b1 )OH, -C(O)-C 3-6 Cycloalkylene-OH, -C(O)-C 1-6 Alkylene-OH, -C(O)-C 3-6 Cycloalkylene-C 1-6 Alkylene -OH, -C(O)OC 1-6 Alkylene-OH, and C optionally substituted by R' 1-6 Alkyl, C 1-6 Alkenyl and C 1-6 Alkynyl; and / or

[0084] Preferably, R 4A Selected from hydrogen, or C 1-6 Alkyl; and / or

[0085] R', R a1 , and R b1 Each occurrence is independently hydrogen, fluorine, chlorine, -OH, C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl, 3, 4, 5 or 6-membered cycloalkyl or 3, 4, 5 or 6-membered heterocyclic group containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, wherein the C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, 3, 4, 5 or 6-membered cycloalkyl, 3, 4, 5 or 6-membered heterocyclyl is unsubstituted or substituted by 1 or 2 independently selected from fluorine, chlorine, -NO2, -CN, -OH, -NH2, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, and hydroxyl C 1-6 The alkylene group is substituted with a substituent.

[0086] In some embodiments, the compound is selected from:

[0087] Wherein R3 is selected from the following groups:

[0088] in Indicates the junction site.

[0089] In some embodiments, the compound is selected from:

[0090] In some embodiments, the compound shown in Formula 1A or a pharmaceutically acceptable salt thereof,

[0091] Wherein, R1 and R2 are each independently selected from hydrogen atom, halogen, C 1-8 Alkyl, C 1-8 alkyl halide;

[0092] A is selected from an oxygen atom or a sulfur atom;

[0093] B is selected from -C(O)- or -P(O)(OH)-;

[0094] R3 is selected from -(C(R 3a )(R 3b )) m -R 3c 、 Among them, when -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m -, the n methylene units are each independently replaced by -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -O-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SO2N(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N- or -N=CH-, and when R3 is When , W1 and W2 are fused together;

[0095] R4 and R8 are single bonds or are independently selected from -O-, C 1-8 Alkylene, C 1-8 Haloalkylene, -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SO2N(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N- or -N=CH-;

[0096] Ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from a 6-10 membered aryl group, a 5-10 membered heteroaryl group, a 3-10 membered cycloalkyl group, or a 3-10 membered heterocyclyl group;

[0097] When ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from a cycloalkyl group or a heterocyclic group, the cycloalkyl group or heterocyclic group is replaced by one or more R w1 、R w2 、R w3 、R w4 、R w5 Replacement, R w1 、R w2 、R w3 、R w4 , or R w5 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 aliphatic groups;

[0098] When ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from an aryl group or a heteroaryl group, the aryl group or heteroaryl group is replaced by one or more R w1 、R w2 , or R w3 、R w4 , or R w5 Replacement, R w1 、R w2 、R w3 、R w4 , or R w5 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(Rb ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 aliphatic groups;

[0099] R6 is selected from hydrogen atom, halogen, C 1-8 Alkyl, C 1-8 Preferably, R6 is selected from a hydrogen atom;

[0100] R7 is selected from C 1-8 Alkylene N(R a )(R b ), C 1-8 Haloalkylene N(R a )(R b ), halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic group; preferably, R7 is selected from C 1-8 Alkylene N(R a )(R b ), or C 1-8 Haloalkylene N(R a )(R b ); Preferably, R7 is selected from -CH2N(R a )(R b );

[0101] Or R6 and R7 can form together with the carbon atom to which they are connected a saturated or unsaturated 5-10 membered cycloalkyl or a 6-10 membered heterocyclic group containing 1-3 heteroatoms independently selected from N, O, and S, wherein the cycloalkyl or heterocyclic group is substituted by one or more independently selected halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R1-6 Preferably, R6 and R7 form a 6-membered cycloalkyl group with the carbon atom to which they are attached, the cycloalkyl group being -N(R a )(R b )replace;

[0102] R 3a , R 3b , R 3c , R5 each occurrence is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 aliphatic groups;

[0103] R, R a , R b Each occurrence is independently hydrogen, protium, deuterium, tritium, halo, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -C(O)CH2OH, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, or C 1-6 aliphatic groups;

[0104] m and n are integers greater than or equal to 0;

[0105] When R3 is -(C(R 3a )(R 3b )) m -R 3c ,-(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m-, the n methylene units are each independently replaced by -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -O-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SON(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N- or -N=CH-;

[0106] Preferably, R3 is -(CH2) a (CH2CH2O) b (CH3) c , or C optionally substituted by R 1-8 Aliphatic group, m=a+3*b+c, b is not 0, a and b are integers greater than 0; when R3 is -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m -n methylene units are each independently replaced by -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -O-, -S-; wherein ring W1, ring W2 and ring W3 are each independently selected from 6-10 membered aryl, 5-8 membered heteroaryl, 4-8 membered cycloalkyl or 4-8 membered heterocyclyl; preferably, ring W1 is selected from phenyl or 4-8 membered heterocyclyl, ring W2 is selected from phenyl or 4-8 membered heterocyclyl, and ring W3 is selected from phenyl or 5-8 membered heteroaryl; preferably, ring W1 is phenyl or oxolanyl,

[0107] Ring W2 is phenyl or oxolanyl, and ring W3 is phenyl or pyridyl;

[0108] When ring W1, ring W2 or ring W3 is selected from cycloalkyl or heterocyclic group, the cycloalkyl or heterocyclic group is replaced by one or more R w1 、R w2 or R w3 Replacement, R w1 、R w2 or R w3 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a)(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 When ring W1, ring W2 or ring W3 is selected from 6-10 membered aryl or 5-8 membered heteroaryl, the aryl or heteroaryl is replaced by one or more R w1 、R w2 or R w3 Replacement, R w1 、R w2 or R w3 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic group; when R3 is -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m -n methylene units are each independently replaced by -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -O-, -S-; wherein ring W1 is selected from 6-10 membered aryl, 4-8 membered cycloalkyl, 4-8 membered heterocyclyl, 5-8 membered heteroaryl, when ring W1 is selected from 4-8 membered cycloalkyl or 4-8 membered heterocyclyl, the cycloalkyl or heterocyclyl is replaced by one or more R w1 Replacement, R w1 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, -(=O), -(=S), -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a)(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 When ring W1 is selected from 6-10 membered aryl or 5-8 membered heteroaryl, the aryl or heteroaryl is replaced by one or more R w1 Replacement, R w1 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 aliphatic groups;

[0109] When R3 is -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m -, the n methylene units of - are each independently replaced by -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -O-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SON(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N-, or -N=CH-;

[0110] Preferably, R3 is m1 is an integer from 1 to 6, and ring W1 is selected from 6-10 membered aryl, 4-8 membered cycloalkyl, 4-8 membered saturated or unsaturated heterocyclic group containing 1-3 heteroatoms selected from N, O, and S, and 5-8 membered heteroaryl containing 1-3 heteroatoms selected from N, O, and S.

[0111] When ring W1 is selected from 4-8 membered cycloalkyl or 4-8 membered heterocyclic group, the cycloalkyl or heterocyclic group is replaced by one or more R w1 Replacement, R w1 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, =O, =S, -NO2, -CN, -OR, -SR, -N(Ra )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R,

[0112] -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 aliphatic groups;

[0113] When ring W1 is selected from 6-10 membered aryl or 5-8 membered heteroaryl, the phenyl or 5-8 membered heteroaryl is replaced by one or more R w1 Replacement, R w1 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 aliphatic groups;

[0114] Preferably, when the ring W1 is surrounded by one or more R w1 When the ring W1 is selected from a 4-8 membered heterocyclic group, it contains 1, 2 or 3 heteroatoms, and each occurrence of the heteroatoms is independently selected from N, O or S; preferably, the ring W1 is

[0115] Ring W1 can be one or more R w1 Replacement, R w1 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(Rb ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 aliphatic groups;

[0116] When R3 is -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m -, wherein the n methylene units are each independently selected from -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -O-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SON(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N- or -N=CH-; wherein ring W1 and ring W2 are each independently selected from substituted or unsubstituted 6-10 membered aryl, 5-8 membered heteroaryl, 4-8 membered cycloalkyl or 4-8 membered heterocyclyl; preferably, ring W1 is ... w1 substituted phenyl or 4-8 membered heterocyclic group, ring W2 is selected from w2 substituted 4-8 membered heterocyclic group; preferably, ring W1 is substituted by one or more R w1 substituted benzene ring or oxolane ring,

[0117] Ring W2 is selected from w2 Replaced

[0118] When ring W1 or ring W2 is selected from a cycloalkyl group or a heterocyclic group, the cycloalkyl group or the heterocyclic group is replaced by one or more R w1 or R w2 Replacement, R w1 or R w2 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 aliphatic groups;

[0119] When ring W1 or ring W2 is selected from 6-10 membered aryl or 5-8 membered heteroaryl, the aryl or heteroaryl is replaced by one or more R w1 or R w2 Replacement, R w1 or R w2 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic group, R4 is a single bond or selected from -O-, C 1-8 Alkylene, C 1-8 Haloalkylene, -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SO2N(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N- or -N=CH-;

[0120] When R3 is -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m -, the n methylene units are each independently replaced by -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -O-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SON(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N- or -N=CH-; Ring W1 and Ring W2 are each independently selected from a 6-10 membered aryl group, a 5-8 membered heteroaryl group, a 4-8 membered cycloalkyl group or a 4-8 membered heterocyclyl group; Preferably, Ring W1 and Ring W2 are fused to form a fused ring structure selected from any of the following

[0121] When ring W1 or ring W2 is a 4-8 membered cycloalkyl group or a 4-8 membered heterocyclic group, it is w1 or R w2 Replacement, R w1 or R w2 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 aliphatic groups;

[0122] When ring W1 or ring W2 is a 6-10 membered aryl group or a 5-8 membered heteroaryl group, the aryl group or heteroaryl group is replaced by one or more R w1 or R w2 Replacement, R w1 or R w2 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 aliphatic groups;

[0123] When R3 is -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m-n methylene units are each independently replaced by -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -O-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SON(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N- or -N=CH-; Rings W1, W2, W3 and W4 are sequentially fused and connected, and Rings W1 and W4 are each independently selected from a 4-8 membered heterocyclic group, which is replaced by one or more R w1 or R w4 Replacement, R w1 or R w4 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic group; Ring W5 and Ring W2 are 6-10 membered aryl or 5-8 membered heteroaryl, the aryl or heteroaryl being one or more R w2 Replacement, R w2 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic group; Ring W3 is a 4-8 membered cycloalkyl group, which is w3 Replacement, R w3 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic group; R4 is a single bond or selected from -O-, C 1-8 Alkylene, C 1-8 Haloalkylene, -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SO2N(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N- or -N=CH-;

[0124] m is an integer from 1 to 20;

[0125] n is an integer from 0 to 8.

[0126] In some embodiments, a compound of Formula 1C or a pharmaceutically acceptable salt thereof,

[0127] Where q is 1, 2 or 3;

[0128] R 1c 、R 2c 、R 5c Each independently selected from hydrogen, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 aliphatic groups;

[0129] or R 1c and R 2cTogether with the carbon atom to which it is attached, it forms a saturated or unsaturated 5-10 membered cycloalkyl or a 6-10 membered heterocyclic group containing 1-3 heteroatoms independently selected from N, O, and S, wherein the cycloalkyl or heterocyclic group is substituted by one or more independently selected halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic group substitution;

[0130] R 6c C 1-8 Alkyl hydroxyl, C 1-8 Alkylamino, -C(R a )(R b )-N(R a )-R 4c 、-C(R a )(R b )-N(R a )-C(O)-R 4c , or -C(R a )(R b )-N(R a )-C(O)OR 4c ;

[0131] R, R a , R b ,R 4c Each occurrence is independently hydrogen, protium, deuterium, tritium, halo, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 aliphatic group, 3-8 membered cycloalkyl group, 3-8 membered heterocyclic group, 6-10 membered aryl group or 5-10 membered heteroaryl group; said C 1-6The aliphatic group, 3-8 membered cycloalkyl group, 3-8 membered heterocyclyl group, 6-10 membered aryl group or 5-10 membered heteroaryl group is unsubstituted or substituted by one or more substituted alkyl radicals independently selected from halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 Aliphatic groups, C 1-6 Alkylene hydroxy substituted.

[0132] The second aspect of the present invention provides a ligand-drug conjugate having a structure as shown in Formula 3 or a pharmaceutically acceptable salt thereof.

[0133] Among them, Ab is the ligand, L is the linker, and D is the drug part;

[0134] n is any integer or decimal from 1 to 15; preferably, n is any integer or decimal from 1 to 13; preferably, n is any integer or decimal from 3 to 10;

[0135] The drug portion D is any of the following structures,

[0136] in,

[0137] R1, R2, R3, R6, R7, A, B, R 1c 、R 2c 、R 5c 、R 1A 、R 2A 、R 3A 、R 4A , and q as defined in any one of claims 2, 3, or 4;

[0138] R 6A Selected from hydrogen, -CH(CH3)N(R 3A )(R 4A ) and -CH2N(R 3A )(R 4A );

[0139] R 5A is hydrogen or C 1-6 alkyl;

[0140] R 1s 、R 2s 、R 7s , and R 7ss Each independently is C 1-8 Alkylene NR-, C 1-8Haloalkylene NR-, -OR, -SR, -NR-, -C(O)NR-, -SO2NR-, or -O-CH2C(O)NR-; preferably, R 1s 、R 2s 、R 7s , and R 7ss Each independently represents -NR-, -NR-CH2-, or -O-CH2C(O)NR-; preferably R 1s 、R 2s 、R 7s , and R 7ss Each independently represents -NH-, -NH-CH2-, or -O-CH2C(O)NH-;

[0141] R 6bs Each occurrence is independently selected from -CH2N(R a1 )-、-CH2N(R a1 )-C 2-5 Alkylene-O-, -CH2N(R a1 )C(O)C(R a1 )(R b1 )O-、-CH2N(R a1 )C(S)C(R a1 )(R b1 )O-、-CH2N(R a1 )C(O)-C 3-8 Cycloalkylene-O-, -CH2N(R a1 )C(O)-C 2-5 Alkylene-O-, -CH2N(R a1 )C(O)C(R a1 )(R b1 )N(R a1 )-、-CH2N(R a1 )C(O)-C 3-8 Cycloalkylene-C(R a1 )(R b1 )-O-、-C 2-5 Alkylene-O-, -CH2N(R a1 )C(O)OC 2-5 Alkylene-O-;

[0142] Among them, R, R a1 , and R b1Each occurrence is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl, 3, 4, 5, 6, 7 or 8-membered cycloalkyl, 3-8 membered heterocyclyl containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, 6-10 membered aryl or 5-10 membered heteroaryl containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, wherein the C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 The alkynyl, 3-, 4-, 5-, 6-, 7-, or 8-membered cycloalkyl, 3- to 8-membered heterocyclyl, 6- to 10-membered aryl, or 5- to 10-membered heteroaryl groups are unsubstituted or substituted with one or more substituted radicals independently selected from halogen, -NO2, -CN, -OH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 Alkyl, C 2-6 Alkenyl or C 1-6 Alkynyl, and hydroxyl C 1-6 The alkylene group is substituted with a substituent; and

[0143] R 6aa Selected from -C 1-8 Alkylene N(R a2 )-、-CH2N(R a2 )C(O)C(R a2 )(R b2 )O-、-CH2N(R a2 )C(O)-C 3-8 Cycloalkylene-O-, -C 2-5 Alkylene-O-, -CH2N(R a2 )C(O)OC 2-5 Alkylene-O-; preferably R 6aa is -CH2-NH2, or -(CH2)4-O-;

[0144] Among them, R a2 and R b2Each occurrence is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, or C 1-6 Alkylene hydroxyl group; preferably, R a2 and R b2 Each occurrence is independently hydrogen, F, Cl, Br, I, -CH2OH, or -C2H4OH.

[0145] In some embodiments, the drug moiety D is selected from any of the following structures,

[0146] in, Indicates the junction site.

[0147] In some embodiments, the linking moiety L is L 1 -L 2 -L 3 -L 4 ;

[0148] L 1 Selected from -(succinimide-3-yl-N)-, -(succinimide-3-yl-N)-WC(=O)-, or

[0149] Wherein, W is selected from C 1-10 Alkylene, C 1-10 Alkylene-cycloalkylene, C 1-10 Heteroalkylene, C 1-10 Alkylene-cycloheteroalkylene, or C 1-10 Heteroalkylene-cycloalkylene, preferably W is C 1-8 Alkylene, C 1-8 Alkylene-cycloalkylene or C 1-8 The heteroalkylene group comprises 1 to 3 heteroatoms independently selected from N, O or S, wherein the alkylene group, cycloalkylene group and heteroalkylene group are unsubstituted or each independently optionally further substituted by one or more substituents selected from halogen, hydroxyl, -CN, amino, alkyl, haloalkyl, deuterated alkyl, alkoxy and cycloalkyl, preferably, the alkylene group, cycloalkylene group and heteroalkylene group are unsubstituted or each independently optionally further substituted by one or more substituents selected from halogen, hydroxyl, -CN, amino, C 1-6 Alkyl, C 1-6 Halogenated alkyl, deuterated C 1-6 Alkyl, C1-6 Alkoxy and C 5-8 More preferably, the alkylene, cycloalkylene and heteroalkylene are unsubstituted or optionally substituted by halogen, -OH, -CN, C 1-3 Alkyl, and -OC 1-3 Alkyl is substituted by a substituent, preferably, the alkylene, cycloalkylene and heteroalkylene are unsubstituted or optionally substituted by a substituent selected from Cl, Br, F, -OH, -CN, methyl, and -OCH3;

[0150] X is selected from a single bond, C 1-10 Alkylene, C 1-10 Alkylene-cycloalkylene, C 1-10 Heteroalkylene, C 1-10 Alkylene-cycloheteroalkylene, or C 1-10 Heteroalkylene-cycloalkylene, preferably X is selected from a single bond, C 1-8 Alkylene, C 1-8 Alkylene-cycloalkylene or C 1-8 heteroalkylene;

[0151] L 2 Selected from -(CH2CH2O) r CH2CH2C(=O)-、-(CH2CH2O) r C(=O)-、-NR 1L (CH2CH2O) r C(=O)- 、 -(CH2CH2O) r CH2C(=O)- 、-NR 1L (CH2CH2O) r CH2CH2C(=O)-、 -NR 1L (CH2CH2O) r CH2C(=O)-、-NR 1L CH2-Ar 1 -(CH2CH2O) r CH2CH2NR 1L C(=O)CH2OCH2C(=O)-、-NR 1L (CH2CH2O) r CH2-Ar 1 -(CH2CH2O) r CH2CH2C(=O)-、-S(CH2) r C(=O)-, -O-(CH2CH2O) r CH2-Ar 1 -(CH2CH2O) r CH2CH2C(=O)-, or a single bond, wherein r is an integer of 12, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, each occurrence of n17 is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30; each occurrence of j is independently selected from 0, 1, 2, 3, 4, 5, 6; preferably, r is an integer of 1, 2, 3, 4, 5, 6, 7, or 8;

[0152] Ar 1 is selected from 6-10 membered aryl, 5-10 membered heteroaryl containing 1-3 heteroatoms independently selected from N, O, P and S, 3-10 membered cycloalkyl or 3-10 membered heterocyclic group containing 1-3 heteroatoms independently selected from N, O, P and S; preferably, Ar 1 Selected from

[0153] L 3 is a peptide residue consisting of 2 to 7 amino acids, preferably L 3 The peptide residue is composed of 2, 3, 4, 5 or 6 amino acids, wherein the amino acids are unsubstituted or optionally further substituted by one or more substituents selected from halogen, hydroxyl, -CN, amino, alkyl, haloalkyl, deuterated alkyl, alkoxy and cycloalkyl, preferably, optionally further substituted by one or more substituents selected from halogen, hydroxyl, -CN, amino, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Alkoxy and C 5-8 substituted by one or more substituents of the cycloalkyl group;

[0154] L 4 Selected from-NR 2L (CR 3L R 4L ) t -Z-(CR 3L R 4L ) t -C(=O)-, -NR 2L (CR 3L R 4L ) t -、 -NR 2L (CR 3L R 4L ) t -Z-(CR 3L R 4L )t -ZC(=O)-, -NR2-Ar 2 -(CR 3L R 4L ) t -ZC(=O)-, or a single bond, wherein t is independently an integer of 0, 1, 2, 3, 4, 5, or 6 at each occurrence; Z is independently a single bond, O, S, or -NH- at each occurrence; Ar 2 is an arylene or heteroarylene group, preferably selected from a 6-membered arylene group or a 5-8-membered heteroarylene group, wherein the heteroarylene group contains 1, 2 or 3 heteroatoms, and the heteroatoms are independently selected from N, O and S; the arylene or heteroarylene group is unsubstituted or optionally selected from H, halogen, -OH, -CN, C 1-6 Alkyl, -OC 1-6 Alkyl, -NH2, -NH(C 1-6 alkyl), and -N(C 1-6 alkyl)2 is substituted with a substituent;

[0155] R 1L and R 2L are the same or different and are independently selected from hydrogen atoms, halogens, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl and -C 1-6 Alkylene-OH;

[0156] R 3L and R 4L are the same or different and are independently selected from hydrogen atoms, halogens, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl and -C 1-6 Alkylene-OH; and

[0157] L 1 The end is connected to the ligand, L 4 The end is connected to the drug portion.

[0158] In some embodiments, L 3 is a peptide residue consisting of 2-6 amino acids selected from glycine, phenylalanine, alanine, valine, lysine, citrulline, serine, glutamic acid, and aspartic acid; preferably a dipeptide residue, a tripeptide residue, or a tetrapeptide residue selected from alanine, phenylalanine, glycine, lysine, and citrulline; preferably, L 3is a peptide residue selected from the group consisting of: glycine-phenylalanine-glycine, alanine-alanine-alanine-glycine, alanine-alanine-alanine, glycine-glycine-phenylalanine-glycine, valine-citrulline, and valine-alanine;

[0159] wherein the peptide residue is unsubstituted or optionally further substituted with one or more substituents selected from halogen, hydroxyl, -CN, amino, alkyl, haloalkyl, deuterated alkyl, alkoxy and cycloalkyl, preferably, optionally further substituted with one or more substituents selected from halogen, hydroxyl, -CN, amino, and C 1-6 Alkyl substituents are substituted.

[0160] In some embodiments, the linking moiety L is selected from:

[0161] in Indicates the junction site.

[0162] In some embodiments, the ligand-drug conjugate is selected from the following structures:

[0163] wherein n is an integer or decimal from 1 to 10; preferably, n is an integer or decimal from 3 to 8; and Ab is a ligand.

[0164] In some embodiments, the Ab is an antibody or an antigen-binding fragment thereof, or a polypeptide, wherein the antibody is selected from a chimeric antibody, a humanized antibody, and a fully human antibody;

[0165] Preferably, the antibody or antigen-binding fragment thereof is selected from anti-TROP-2 antibody, anti-HER2 (ErbB2) antibody, anti-NECTIN4 antibody, anti-EGFR antibody, anti-B7-H3 antibody, anti-c-Met antibody, anti-HER3 (ErbB3) antibody, anti-HER4 (ErbB4) antibody, anti-LIV-1 antibody, anti-ROR1 antibody, anti-CD20 antibody, anti-CD22 antibody, anti-CD30 antibody, anti-CD33 antibody, anti-CD44 antibody, anti-CD56 antibody, anti-CD70 antibody, anti-CD73 antibody, anti-CD105 antibody, anti-CEA antibody, anti-A33 antibody, anti-Cripto antibody, anti-EphA2 antibody, anti-G250 antibody, anti-MUCl antibody, anti-Lewis Y antibody, anti-VEGFR antibody, anti-GPNMB antibody, anti-Integrin antibody, anti-PSMA antibody, anti-Tenascin-C antibody, anti-SLC44A4 antibody, anti-Mesothelin antibody or antigen-binding fragment thereof;

[0166] Preferably, the antibody or antigen-binding fragment thereof is an anti-TROP-2 antibody, an anti-NECTIN4 antibody, an anti-B7-H3 antibody, an anti-HER2 (ErbB2) antibody, an anti-HER3 (ErbB3) antibody, an anti-LIV-1 antibody, an anti-ROR1 antibody or an antigen-binding fragment thereof;

[0167] Preferably, the antibody or antigen-binding fragment thereof is an anti-HER2 (ErbB2) antibody, an anti-NECTIN4 antibody, an anti-B7-H3 antibody or an antigen-binding fragment thereof;

[0168] Preferably, the antibody or antigen-binding fragment thereof is trastuzumab, ifinatamab monoclonal antibody or PADCEV monoclonal antibody.

[0169] The third aspect of the present invention provides a compound, such as a compound shown in Formula 5 or a pharmaceutically acceptable salt thereof,

[0170] L j -L2-L3–L4-D (Formula 5);

[0171] wherein -L2-, -L3-, -L4-, and -D are as defined in the second aspect of the present invention;

[0172] L j Selected from and W and X are as defined in any one of the second aspect of the present invention.

[0173] In some embodiments, the compound is selected from:

[0174] The fourth aspect of the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the compound or ligand-drug conjugate according to any one of the first, second or third aspects, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0175] The fifth aspect of the present invention provides a pharmaceutical composition, comprising the use of the compound or ligand-drug conjugate or pharmaceutically acceptable salt thereof according to any one of the first, second or third aspects of the present invention, or the pharmaceutical composition according to the fourth aspect, in the preparation of a medicament for treating or preventing tumors;

[0176] Preferably, the tumor is a cancer associated with B7-H3 expression, HER2 expression, or NECTIN4 expression;

[0177] Preferably, the cancer is selected from breast cancer, gastric cancer, melanoma and lung cancer.

[0178] The sixth aspect of the present invention provides a method for preventing or treating tumors, comprising administering to a subject in need thereof an effective amount of the compound or ligand-drug conjugate or a pharmaceutically acceptable salt thereof according to any one of the first, second, or third aspects of the present invention, or the pharmaceutical composition according to the fourth aspect;

[0179] Preferably, the tumor is a cancer associated with B7-H3 expression, HER2 expression, or NECTIN4 expression;

[0180] Preferably, the cancer is selected from breast cancer, gastric cancer, melanoma and lung cancer.

[0181] The sixth aspect of the present invention provides a compound or ligand-drug conjugate or a pharmaceutically acceptable salt thereof according to any one of the first, second or third aspects, or the pharmaceutical composition according to the fourth aspect, for use in preventing or treating tumors;

[0182] Preferably, the tumor is a cancer associated with B7-H3 expression, HER2 expression or NECTIN4 expression; preferably, the cancer is selected from breast cancer, gastric cancer, melanoma and lung cancer.

[0183] The sixth aspect of the present invention provides a use of the compound described in the first aspect of the present invention, characterized in that it is used as a toxin in an antibody-drug conjugate to prepare an antibody-drug conjugate. BRIEF DESCRIPTION OF THE DRAWINGS

[0184] FIG1 shows the inhibitory activity of B7H3-ADC-27 in the B7H3-negative Raji cell line in Experimental Example 6;

[0185] FIG2 shows the inhibitory activity of B7H3-ADC-27 in a mixed B7H3-positive A375 and negative Raji cell line in Experimental Example 6;

[0186] FIG3 shows the inhibitory activity of B7H3-ADC-1 in the B7H3-negative Raji cell line in Experimental Example 6;

[0187] FIG4 shows the inhibitory activity of B7H3-ADC-1 in a mixed B7H3-positive A375 and negative Raji cell line in Experimental Example 6;

[0188] FIG5 shows the tumor inhibitory activity of the ADC molecule in Experimental Example 6 in an animal model in which both B7H3-negative and -positive cells coexist;

[0189] FIG6 is a tumor growth curve of mice in Experimental Example 7;

[0190] FIG7 is a curve showing changes in body weight of mice in Experimental Example 7;

[0191] FIG8 is a tumor growth curve of mice in Experimental Example 8;

[0192] FIG9 is a curve showing changes in body weight of mice in Experimental Example 8;

[0193] Figure 10 is a curve showing the weight changes of mice in Experimental Example 9;

[0194] FIG11 is a tumor growth curve of mice in Experimental Example 10;

[0195] FIG12 shows the tumor tissues of mice in Experimental Example 10. DETAILED DESCRIPTION

[0196] The following is a clear and complete description of the technical solution of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0197] Terms and Definitions

[0198] Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meanings as commonly understood by one of ordinary skill in the art.

[0199] Certain compounds of the present invention may exist in free form or, where appropriate, in the form of pharmaceutically acceptable derivatives thereof. In the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, prodrugs, stereoisomers (including but not limited to diastereomers and enantiomers), tautomers, solvates, polymorphs and isotopic compounds, which, after being administered to a patient in need thereof, can directly or indirectly provide a compound of the present invention or a metabolite thereof. Therefore, when referring to a "compound of the present invention" herein, it is also intended to encompass the various derivative forms of the compound described above.

[0200] The term "pharmaceutically acceptable salt" refers to a salt that retains the biological effectiveness of the free acids and bases of the specified compound without any adverse biological effects. Examples of pharmaceutically acceptable salts include, but are not limited to: (1) acid addition salts, such as salts formed with inorganic acids such as hydrochloric acid, sulfuric acid, hydrobromic acid, nitric acid, phosphoric acid, etc.; or salts formed with organic acids such as malic acid, fumaric acid, maleic acid, benzoic acid, phenylacetic acid, succinic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, glycolic acid, cinnamic acid, pyruvic acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, acrylic acid, mandelic acid, etc.; or (2) base addition salts, such as salts formed with alkali metals such as lithium, sodium, potassium, etc.; and salts formed with alkaline earth metals such as calcium, magnesium, etc.; and salts formed with organic bases such as ammonium, choline, diethanolamine, lysine, ethylenediamine, tert-butylamine, tert-octylamine, tris(hydroxymethyl)aminomethane, N-methylglucamine, triethanolamine, dehydroabietinamine, etc. Other pharmaceutically acceptable salts are known to those skilled in the art.

[0201] Prodrugs of the compounds of the present invention are included within the scope of protection of the present invention. Generally, prodrugs are functional derivatives that are easily converted into the desired compound in vivo. Therefore, the term "administering" in the treatment methods provided herein includes administering the compounds disclosed herein, or methods that, although not explicitly disclosed, can be converted into the compounds disclosed herein in vivo after administration to a subject to treat the various diseases described. Conventional methods for selecting and preparing suitable prodrug derivatives are described in books such as "Design of Prodrugs" (H. Bundgaard, Elsevier, 1985).

[0202] The compounds of the present invention may contain one or more asymmetric centers and may thus produce diastereomers and optical isomers. The present invention includes all possible diastereomers and racemic mixtures thereof, their substantially pure resolved enantiomers, all possible geometric isomers and pharmaceutically acceptable salts thereof.

[0203] The compounds of the present invention do not have a precise stereostructure at any particular position within the compounds. The present invention encompasses all stereoisomers of the compounds and pharmaceutically acceptable salts thereof. Furthermore, mixtures of stereoisomers and isolated specific stereoisomers are also encompassed by the present invention. During the synthetic process for preparing such compounds, or using racemization or epimerization methods known to those skilled in the art, the resulting products may be mixtures of stereoisomers.

[0204] When the compounds of the present invention exist in tautomers, unless otherwise stated, the present invention includes any possible tautomers and pharmaceutically acceptable salts thereof, and mixtures thereof.

[0205] When the compounds of the present invention and their pharmaceutically acceptable salts exist in the form of solvates or polymorphs, the present invention includes any possible solvates and polymorphs. The type of solvent used to form the solvate is not particularly limited, as long as the solvent is pharmacologically acceptable. For example, water, ethanol, propanol, acetone and the like can be used.

[0206] The present invention also includes all pharmaceutically acceptable isotopic compounds that are identical to the compounds of the present invention except that one or more atoms are replaced by an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number prevalent in nature. Examples of suitable isotopes for inclusion in the compounds of the present invention include, but are not limited to, isotopes of hydrogen (e.g., deuterium (2H), tritium (3H)); isotopes of carbon (e.g., 13C and 14C); isotopes of chlorine (e.g., 37Cl); isotopes of iodine (e.g., 125I); isotopes of nitrogen (e.g., 13N and 15N); isotopes of oxygen (e.g., 17O and 18O); isotopes of phosphorus (e.g., 32P); and isotopes of sulfur (e.g., 34S).

[0207] The term "ligand" refers to a macromolecular compound that recognizes and binds to an antigen or receptor associated with a target cell. The function of a ligand is to present a drug to the target cell population bound to the ligand. In embodiments of the present invention, the ligand is represented by an Ab. The ligand can form a bond with a linker through a heteroatom on the ligand. The ligand is preferably an antibody, an antigen-binding fragment thereof, or a polypeptide. The antibody is selected from a chimeric antibody, a humanized antibody, a fully human antibody, or a murine antibody; preferably, a monoclonal antibody.

[0208] The term "ligand binding component" refers to a group capable of attaching to a ligand moiety.

[0209] The term "drug linking component" refers to a group capable of linking to a drug moiety.

[0210] The term "drug" refers to a cytotoxic drug, represented by D, which is a chemical molecule that has a strong ability to disrupt the normal growth of tumor cells.

[0211] The term "linker unit" or "linking fragment" or "linking unit" or "linking portion" or "linker" refers to a chemical structure fragment or bond that is connected to a ligand at one end and to a drug at the other end, and can also be connected to other linkers before being connected to the drug.

[0212] The term "ligand-drug conjugate" refers to a ligand linked to a biologically active drug via a stable linker. In the present disclosure, a "ligand-drug conjugate" is preferably an antibody-drug conjugate (ADC), which refers to a monoclonal antibody or antibody fragment linked to a biologically active toxic drug via a stable linker.

[0213] The three letter and one letter codes for amino acids used in this disclosure are as described in J. biol. chem, 243, p3558 (1968).

[0214] The term "antibody" refers to immunoglobulins, which are tetrapeptide chains composed of two identical heavy chains and two identical light chains connected by interchain disulfide bonds. The amino acid composition and order of the constant region of immunoglobulins' heavy chains vary, resulting in different antigenicity. Consequently, immunoglobulins can be divided into five classes, or isotypes, namely IgM, IgD, IgG, IgA, and IgE, with their corresponding heavy chains being μ, δ, γ, α, and ε, respectively. Within the same class, Igs are further divided into subclasses based on the amino acid composition of their hinge regions and the number and location of heavy chain disulfide bonds. For example, IgG can be divided into IgG1, IgG2, IgG3, and IgG4. Light chains are classified as either kappa or lambda chains based on differences in their constant regions. Each of the five Ig classes can have either kappa or lambda chains. The antibodies disclosed herein are preferably specific antibodies against cell surface antigens on target cells, and non-limiting examples thereof include the following antibodies: anti-TROP-2 antibody, anti-HER2 (ErbB2) antibody, anti-EGFR antibody, anti-NECTIN4 antibody, anti-B7-H3 antibody, anti-c-Met antibody, anti-HER3 (ErbB3) antibody, anti-HER4 (ErbB4) antibody, anti-LIV-1 antibody, anti-ROR1 antibody, anti-CD20 antibody, anti-CD22 antibody, anti-CD30 antibody, anti-CD33 antibody, anti-CD44 antibody, anti-CD56 antibody, anti-CD70 antibody, anti-CD73 antibody, anti-CD105 antibody, anti-CEA antibody, anti-A33 antibody, anti-Cripto antibody, anti-EphA2 antibody, anti-G250 antibody, anti-MUCl antibody, anti-Lewis antibody. Y antibody, anti-VEGFR antibody, anti-GPNMB antibody, anti-Integrin antibody, anti-PSMA antibody, anti-Tenascin-C antibody, anti-SLC44A4 antibody, anti-Mesothelin antibody or an antigen-binding fragment thereof; more preferably, the antibody or antigen-binding fragment thereof is an anti-B7-H3 antibody, anti-NECTIN4 antibody, anti-HER2 (ErbB2) antibody or an antigen-binding fragment thereof.

[0215] The antibodies of the present invention include murine antibodies, chimeric antibodies, humanized antibodies and fully human antibodies, with humanized antibodies and fully human antibodies being preferred.

[0216] The term "murine antibody" as used herein refers to antibodies produced in mice according to the knowledge and skills in the art. During production, a test subject is injected with a specific antigen and then a hybridoma expressing an antibody with the desired sequence or functional properties is isolated.

[0217] The term "chimeric antibody" refers to an antibody created by fusing the variable region of a mouse antibody with the constant region of a human antibody, which can mitigate the immune response induced by the mouse antibody. To create a chimeric antibody, one must first establish a hybridoma that secretes mouse-specific monoclonal antibodies. The variable region genes are then cloned from the mouse hybridoma cells. Furthermore, the constant region genes of the human antibody are cloned as needed. The mouse variable region genes and the human constant region genes are then linked to form a chimeric gene, which is then inserted into an expression vector. Finally, the chimeric antibody molecule is expressed in a eukaryotic or prokaryotic system.

[0218] The term "humanized antibody," also known as a CDR-grafted antibody, refers to an antibody produced by grafting murine CDR sequences onto a human variable region framework, i.e., different types of human germline antibody framework sequences. This overcomes the xenobiotic response induced by chimeric antibodies due to the presence of a large amount of murine protein.

[0219] The terms "fully human antibody", "fully human antibody" or "completely human antibody", also known as "fully human monoclonal antibody", refer to antibodies whose variable and constant regions are both human, eliminating immunogenicity and toxic side effects.

[0220] The term "antigen-binding fragment" refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen. It has been shown that fragments of a full-length antibody can be used to perform the antigen-binding function of an antibody. Examples of binding fragments included in "antigen-binding fragments" include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL, and CH1 domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments connected by a disulfide bridge on the hinge region, (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) an Fv fragment consisting of the VH and VL domains of a single arm of an antibody; (v) a single domain or dAb fragment (Ward et al., (1989) Nature 341: 544-546), which consists of a VH domain; and (vi) isolated complementarity determining regions (CDRs) or (vii) a combination of two or more isolated CDRs, optionally connected by a synthetic linker. In addition, although the two domains VL and VH of the Fv fragment are encoded by separate genes, they can be connected by synthetic linkers using recombinant methods, so that they can be produced as a single protein chain in which the VL and VH regions are paired to form a monovalent molecule (called single-chain Fv (scFv); see, for example, Bird et al. (1988) Science 242: 423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci USA 85: 5879-5883). Such single-chain antibodies are also intended to be included in the term "antigen-binding fragment" of an antibody. Such antibody fragments are obtained using conventional techniques known to those skilled in the art, and the fragments are screened for functionality in the same manner as for intact antibodies. Antigen-binding portions can be produced by recombinant DNA technology or by enzymatic or chemical fragmentation of intact immunoglobulins.

[0221] Fab is an antibody fragment having a molecular weight of approximately 50,000 and antigen-binding activity, among fragments obtained by treating IgG antibody molecules with the protease papain (cleaving the amino acid residue at position 224 of the H chain), in which approximately half of the N-terminal side of the H chain and the entire L chain are bound together by a disulfide bond.

[0222] F(ab')2 is an antibody fragment having a molecular weight of about 100,000 and antigen-binding activity, obtained by digesting the portion below the two disulfide bonds in the hinge region of IgG with the enzyme pepsin, and comprises two Fab regions linked at the hinge position.

[0223] Fab' is an antibody fragment having a molecular weight of about 50,000 and antigen-binding activity, obtained by cleaving the disulfide bond of the hinge region of the above-mentioned F(ab')2.

[0224] Furthermore, the Fab' fragment of the antibody can be produced by inserting a DNA encoding the Fab' fragment into a prokaryotic expression vector or a eukaryotic expression vector and introducing the vector into a prokaryotic or eukaryotic organism to express the Fab'.

[0225] The term "single-chain antibody", "single-chain Fv" or "scFv" refers to a molecule comprising an antibody heavy chain variable domain (or region; VH) and an antibody light chain variable domain (or region; VL) connected by a linker. Such scFv molecules can have the general structure: NH2-VL-linker-VH-COOH or NH2-VH-linker-VL-COOH. Suitable prior art linkers consist of repeated GGGGS amino acid sequences or variants thereof, for example, variants using 1-4 repeats (Holliger et al. (1993), Proc. Natl. Acad. Sci. USA 90: 6444-6448). Other linkers useful in the present disclosure are described by Alfthan et al. (1995), Protein Eng. 8:725-731, Choi et al. (2001), Eur. J. Immunol. 31:94-106, Hu et al. (1996), Cancer Res. 56:3055-3061, Kipriyanov et al. (1999), J. Mol. Biol. 293:41-56, and Roovers et al. (2001), Cancer Immunol.

[0226] The term "CDR" refers to one of the six hypervariable regions in the variable domain of an antibody that primarily contribute to antigen binding. One of the most commonly used definitions of the six CDRs is provided by Kabat EA et al. (1991) Sequences of proteins of immunological interest. NIH Publication 91-3242). As used herein, the Kabat definition of CDR is only applied to CDR1, CDR2, and CDR3 (CDR L1, CDR L2, CDR L3, or L1, L2, L3) of the light chain variable domain, and CDR2 and CDR3 (CDR H2, CDR H3, or H2, H3) of the heavy chain variable domain.

[0227] The term "vector" refers to a nucleic acid molecule capable of transporting another nucleic acid connected thereto. In one embodiment, a vector is a "plasmid", which refers to a circular double-stranded DNA loop into which another DNA segment can be connected. In another embodiment, a vector is a viral vector, in which another DNA segment can be connected to a viral genome. Vectors disclosed herein can autonomously replicate in the host cell into which they have been introduced (e.g., bacterial vectors and additional mammalian vectors with a bacterial origin of replication) or can be integrated into the genome of the host cell after introducing the host cell, thereby replicating (e.g., non-additional mammalian vectors) with the host genome.

[0228] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 carbon atoms, more preferably an alkyl group containing 1 to 10 carbon atoms, and most preferably an alkyl group containing 1 to 6 carbon atoms. The term "C1-6 alkyl" refers to a saturated straight or branched chain hydrocarbon group having 1 to 6 carbon atoms (e.g., 1, 2, 3, 4, 5, or 6 carbon atoms). For example, "C1-6 alkyl" can be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, or n-hexyl, etc.

[0229] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 10 carbon atoms, and most preferably 3 to 8 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; polycyclic cycloalkyls include spirocyclic, fused, and bridged cycloalkyls.

[0230] The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 20 ring atoms, wherein one or more (e.g., 1, 2, or 3) ring atoms are selected from nitrogen, oxygen, or sulfur, and the remaining ring atoms are carbon. Preferably, it contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably, the cycloalkyl ring contains 3 to 10 ring atoms. Non-limiting examples of monocyclic heterocyclyls include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and the like. Polycyclic heterocyclyls include spirocyclic, fused, and bridged heterocyclyls.

[0231] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings sharing adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6- to 10-membered, such as phenyl and naphthyl, with phenyl being preferred. The aryl ring may be fused to a heteroaryl, heterocyclyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the aryl ring.

[0232] The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur, and nitrogen. The heteroaryl group is preferably 5- to 10-membered, more preferably 5- or 6-membered, such as furanyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, and the like. The heteroaryl ring may be fused to an aryl, heterocyclyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the heteroaryl ring.

[0233] The term "haloalkyl" refers to an alkyl group substituted with one or more halo groups.

[0234] The term "deuterated alkyl" refers to an alkyl group substituted with one or more deuterium atoms.

[0235] The term "halo" refers to fluoro, chloro, bromo or iodo.

[0236] The term "drug moiety" or "D" generally refers to any compound that has the desired biological activity and a reactive functional group for preparing the conjugates of the present invention.

[0237] The term "cytotoxic compound" refers to a class of drugs that can effectively kill tumor cells and inhibit their proliferation. These include topoisomerase I (TOP1) inhibitors, tubulin polymerization inhibitors, topoisomerase II (TOP2) inhibitors, dihydrofolate reductase inhibitors, thymidine synthetase inhibitors, purine nucleoside synthetase inhibitors, ribonucleotide reductase inhibitors, DNA polymerase inhibitors, RNA polymerase II inhibitors, and other compounds that can inhibit cell proliferation. Topoisomerase I (TOP1) inhibitors include, but are not limited to, camptothecin derivatives, such as SN-38, Dxd, and Dx-8951, and tubulin polymerization inhibitors, such as Eribulin, MMAE, MMAF, and maytansine (structures shown below).

[0238] The term "camptothecin derivatives" refers to pyrroloquinoline alkaloid derivatives, such as wait.

[0239] The terms "substituted" and "substituted" mean that one or more (e.g., one, two, three, or four) hydrogen atoms on the designated atom are replaced with a group selected from the indicated group, provided that the designated atom's normal valency in the present context is not exceeded and that the substitution results in a stable compound. Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0240] If a substituent is described as "optionally substituted with...", the substituent may be (1) unsubstituted, or (2) substituted. If an atom or group is described as optionally substituted with one or more of the substituents listed, one or more hydrogen atoms on the atom or group may be replaced with independently selected, optional substituents. If a substituent is described as "independently selected from" or "each independently is," each substituent is selected independently of the others. Thus, each substituent may be the same as or different from another substituent. For example, when a substituent or substitution position or different substituents or substitution positions have R groups (such as, but not limited to, R2, R3, Rh, Ri, Rx, and / or Ry) that may be designated by the same or different symbols, each R is selected independently of the others, i.e., may be the same or different. The same is true for the selection of numerical values ​​such as d, g, m, and n.

[0241] Unless otherwise indicated, as used herein, the point of attachment of a substituent may be from any suitable position of the substituent.

[0242] When a bond to a substituent is shown to pass through a bond connecting two atoms in a ring, then such substituent may be bonded to any ring atom in the substitutable ring.

[0243] The terms "include," "comprising," "having," "containing," or "involving," and their variations herein, are inclusive or open-ended and do not exclude other unrecited elements or method steps. Those skilled in the art will understand that the above terms, such as "comprising," encompass the meaning of "consisting of."

[0244] As used herein, a "pharmaceutically acceptable carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the active ingredient is administered and which is suitable, within the scope of sound medical judgment, for contact with the tissues of humans and / or other animals without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit / risk ratio. The terms "active ingredient," "therapeutic agent," "active substance," or "active agent" refer to a chemical entity that is effective in treating one or more symptoms of a target disorder or condition.

[0245] As used herein, the term "effective amount" (e.g., "therapeutically effective amount" or "prophylactically effective amount") refers to an amount of active ingredient that, after administration, will achieve the desired effect to some extent, such as alleviating one or more symptoms of the condition being treated or preventing the appearance of the condition or its symptoms.

[0246] As used herein, unless otherwise indicated, the term "treat," ...

[0247] Example

[0248] The experimental methods in the following examples are conventional methods unless otherwise specified. The chemical raw materials, reagents, etc. used in the following examples are commercially available products unless otherwise specified. The abbreviations and their meanings appearing in this article are as follows:

[0249] Table 1 Abbreviations and their meanings

[0250] Cell name and origin

[0251] Unless otherwise stated, the cell strains or cell lines used in the examples of the present invention can be obtained through commercial channels.

[0252] SK-BR-3 cell line was purchased from Nanjing Beiruiji Biotechnology Co., Ltd.;

[0253] NCI-N87 cell line was purchased from Nanjing Beiruiji Biotechnology Co., Ltd.;

[0254] T47D cell line was purchased from Nanjing Kebai Biotechnology Co., Ltd.

[0255] MDA-MB-453 cell line was purchased from Nanjing Kebai Biotechnology Co., Ltd.

[0256] A375 cell line was purchased from ATCC;

[0257] NCI-H1703 cell line was purchased from Nanjing Kebai Biotechnology Co., Ltd.

[0258] Example 1:

[0259] Synthesis route:

[0260] Synthesis of intermediate 1-2:

[0261] Under an ice-water bath, 20 mL of a 33% HBr solution in AcOH was added to a single-necked flask. 1,2,3,4-tetra-O-acetyl-β-D-glucuronic acid methyl ester (1-1.5 g, 13.2 mmol) was slowly added to the system. After stirring for 10 minutes, the mixture was warmed to room temperature. Stirring overnight, the reaction solution gradually clarified. After completion of the reaction, DCM was added to the reaction system for dilution. The mixture was extracted three times with water (50 mL x 3). The organic phases were then washed three times with saturated aqueous NaHCO₃ and aqueous NaCl. The combined organic phases were dried over anhydrous Na₂SO₄ and concentrated to yield compound 1-2 (4.3 g, 82%).

[0262] Synthesis of intermediate 1-3:

[0263] Compound 1-2 (4.9 g, 12.3 mmol) was dissolved in 30 mL of acetonitrile, and 4-hydroxybenzaldehyde (1.8 g, 14.9 mmol) and silver oxide (4.2 g, 18.1 mmol) were added sequentially, stirring overnight. TLC monitoring indicated the reaction was complete, and the mixture was filtered through celite and washed with ethyl acetate. After spin drying and concentration, the crude product was redissolved in ethyl acetate and washed three times with saturated aqueous NaHCO₃ and aqueous NaCl. The organic phases were combined, dried over anhydrous Na₂SO₄, concentrated, and separated by column chromatography (petroleum ether:ethyl acetate = 4:1 to 2:1) to obtain compound 1-3 (2.7 g, 50% yield). MS (ESI): m / z found [M+18]+ = 456.1.

[0264] Synthesis of intermediate 1-4:

[0265] Compound 1-3 (6 g, 13.6 mmol) was dissolved in dry THF and NaBH₄ (1 g, 27.2 mmol) was added at 0°C. After 1 h of reaction, TLC monitoring indicated complete reaction. The reaction mixture was diluted with DCM and washed with saturated aqueous NaCl (200 mL x 4). The organic phases were combined, dried over anhydrous Na₂SO₄, and concentrated to afford compound 1-4 (5 g, 83%).

[0266] Synthesis of intermediate 1-5:

[0267] Compound 1-4 (63 mg, 0.143 mmol) was dissolved in DCM under ice-water bath conditions. p-Nitrophenyl chloroformate (29 mg, 0.143 mmol) and triethylamine (40 μL, 0.286 mmol) were added sequentially. The mixture was allowed to react in the ice-water bath for 10 minutes, then warmed to room temperature and stirred for 1 hour. TLC monitoring indicated that compound 1-4 had reacted to no change, and the reaction solution was retained for later use.

[0268] Synthesis of intermediate 1-7:

[0269] Under an ice-water bath, isotecan mesylate (1-6, 500 mg, 0.941 mmol) was dissolved in DMF. Di-tert-butyl dicarbonate (226 mg, 1.035 mmol) and triethylamine (262 μL, 1.881 mmol) were added sequentially. After stirring at 0°C for 15 minutes, the mixture was allowed to warm to room temperature and reacted overnight. TLC monitoring indicated the reaction was complete. The reaction solution was diluted with ethyl acetate, washed three times with water, and the combined organic phases were dried over anhydrous Na2SO4 and concentrated to afford compound 1-7 (600 mg, 100%).

[0270] Synthesis of intermediate 1-8:

[0271] Under an ice-water bath, 1-7 (100 mg, 0.187 mmol) was dissolved in DCM, and p-nitrophenyl chloroformate (151 mg, 0.747 mmol) and 4-dimethylaminopyridine (91 mg, 0.747 mmol) were added sequentially. The reaction was allowed to react at 0°C for 1 hour. After TLC monitoring of the reaction completion, 0.1N aqueous hydrochloric acid was added, followed by washing and extraction. The organic phase was washed three times with water and once with saturated aqueous NaCl, dried over anhydrous NaSO, concentrated, and purified by PTLC to afford 1-8 (110 mg, 84%).

[0272] Synthesis of intermediate 1-9:

[0273] Under ice-water bath, 1-8 (110 mg, 0.157 mmol) was dissolved in DMF, and N,N-dimethylethylenediamine (13.85 mg, 0.157 mmol) was added and reacted at 0°C for 10 minutes. The reaction was completed after TLC monitoring. The reaction solution was diluted with ethyl acetate, washed three times with water, and the combined organic phases were dried over anhydrous Na2SO4 and concentrated. After PTLC purification, compound 1-9 (62 mg, 75%) was obtained.

[0274] Synthesis of intermediate 1-10:

[0275] 1-9 (62 mg, 0.143 mmol) was added to the reaction mixture of 1-5 at room temperature and allowed to react overnight. TLC monitored the reaction completion. The organic phase was washed three times with water and once with saturated aqueous NaCl, dried over anhydrous Na2SO4, concentrated, and purified by P-TLC to afford 1-10 (50 mg, 45%).

[0276] Synthesis of intermediate 1-11:

[0277] Under an ice-water bath, 1-10 (30 mg, 0.027 mmol) was dissolved in ultra-dry DCM (5 mL) and trifluoroacetic acid (0.5 mL) was added for half an hour. TLC monitored the reaction completion. The reaction solution was diluted with ethyl acetate and neutralized with saturated sodium bicarbonate solution. The combined organic phases were washed three times with water, dried over anhydrous Na2SO4, and concentrated to afford compound 1-11 (27 mg, 98%).

[0278] Synthesis of compound 1:

[0279] At room temperature, 1-11 (36 mg, 0.035 mmol) was dissolved in methanol, and sodium methoxide (0.15 mg, 0.003 mmol) was added and reacted at room temperature. The reaction was monitored to be complete by TLC. The mixture was placed in an ice-water bath, and 2N sodium hydroxide solution (2.2 mg, 0.054 mmol) was added and reacted at 0°C for one hour. The reaction was monitored to be complete by TLC. The reaction solution was neutralized to neutrality by adding 1N hydrochloric acid, concentrated, and purified by HPLC to give compound 1 (15 mg, 48%). MS (ESI): m / z found [M+H]+=876.4.

[0280] Example 2:

[0281] Synthesis route:

[0282] Synthesis of intermediate 2-1:

[0283] At room temperature, compound 1-7 (50 mg, 0.093 mmol) was dissolved in 5 mL of ultra-dry dichloromethane. 4-Methyl-1-piperazinepropionic acid (32 mg, 0.182 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (80 mg, 0.449 mmol), and 4-dimethylaminopyridine (8 mg, 0.060 mmol) were added sequentially. The mixture was allowed to react overnight. LCMS monitoring indicated the reaction was complete. The reaction solution was concentrated and purified by PTLC to afford compound 2-1 (40 mg, 63%). MS (ESI): m / z found [M+H]+ = 690.3.

[0284] Synthesis of compound 2:

[0285] Under an ice-water bath, 2-1 (40 mg, 0.019 mmol) was dissolved in ultra-dry dichloromethane (5 mL) and trifluoroacetic acid (0.5 mL) was added for 1 hour. The reaction was complete after LCMS monitoring. The reaction solution was diluted with ethyl acetate and extracted with saturated sodium bicarbonate solution. The product was washed with saturated aqueous NaCl solution, dried over anhydrous Na2SO4, and concentrated. Pre-HPLC purification afforded compound 2 (0.79 mg, 7%). MS (ESI): m / z found [M+H]+ = 590.3.

[0286] Example 3:

[0287] Synthesis route:

[0288] Synthesis of intermediate 3-2:

[0289] At room temperature, p-hydroxyphenylacetic acid (1.0 g, 6.57 mmol) was dissolved in DMF (30 mL). Benzyl bromide (1.1 g, 6.57 mmol) and potassium carbonate (0.9 g, 6.57 mmol) were added sequentially and allowed to react overnight. LCMS monitoring indicated the reaction was complete. The reaction solution was diluted with ethyl acetate and extracted with water. The mixture was washed with saturated aqueous NaCl (200 mL x 4). The organic phases were combined, dried over anhydrous Na2SO4, concentrated, and purified by normal phase column chromatography to yield compound 3-2 (1.7 g, 100%). MS (ESI): m / z found [M+18]. + =260.2.

[0290] Synthesis of intermediate 3-3:

[0291] Under an ice-water bath, 3-2 (1.0 g, 4.128 mmol) was dissolved in DCM (50 mL). After shielding with tin foil, silver trifluoromethanesulfonate (1.54 g, 6.004 mmol) and 2,3,4,6-tetraacetoxy-α-D-pyranose glucopyranose bromide (2.5 g, 6.004 mmol) were added sequentially and reacted at 0°C for 2 hours. The reaction was monitored for completion by TLC. The reaction solution was quenched by N,N-diisopropylethylamine, and the filtrate was filtered, concentrated, and purified by normal phase column chromatography to afford 3-3 (1.5 g, 62%). MS (ESI): m / z found [M+18]+ = 590.2.

[0292] Synthesis of intermediate 3-4:

[0293] At room temperature, 3-3 (1.5 g, 2.56 mmol) was dissolved in ultra-dry ethyl acetate (10 mL) and methanol (2 mL). After hydrogen displacement with an appropriate amount of palladium on carbon, the reaction was allowed to react for 1 hour. The reaction was monitored by TLC. The reaction solution was filtered, the filtrate was concentrated, and purified by normal phase column chromatography to afford compound 3-4 (678 mg, 55%).

[0294] Synthesis of intermediate 3-5:

[0295] At room temperature, 1-7 (50 mg, 0.093 mmol) was dissolved in dichloromethane (5 mL), and compound 3-4 (88 mg, 0.182 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (87 mg, 0.449 mmol), and 4-dimethylaminopyridine (8 mg, 0.060 mmol) were added in sequence. The reaction was allowed to react overnight. LCMS monitoring showed that the reaction was complete. The reaction solution was concentrated and purified by PTLC to give compound 3-5 (72 mg, 77%). MS (ESI): m / z found [M+H] + =1000.4.

[0296] Synthesis of intermediate 3-6:

[0297] Under an ice-water bath, 3-5 (72 mg, 0.072 mmol) was dissolved in ultra-dry dichloromethane (5 mL) and trifluoroacetic acid (0.5 mL) was added for one hour. LCMS monitored the reaction for completion. The reaction mixture was diluted with ethyl acetate and extracted with saturated sodium bicarbonate solution. The mixture was washed with saturated aqueous NaCl solution, dried over anhydrous Na2SO4, and concentrated to afford compound 3-6 (90 mg, 100%). MS (ESI): m / z found [M+H]+ = 900.3.

[0298] Synthesis of compound 3:

[0299] At room temperature, 3-6 (32 mg, 0.036 mmol) was dissolved in methanol (5 mL), and sodium carbonate (23 mg, 0.216 mmol) was added. The mixture was allowed to react overnight at room temperature. The reaction was complete after LCMS monitoring. The reaction solution was filtered, and the filtrate was concentrated and purified by Pre-HPLC to obtain compound 3 (1.6 mg, 6%). MS (ESI): m / z found [M+H] + =734.1.

[0300] Example 4:

[0301] Synthesis of intermediate 4-1:

[0302] At room temperature, 1-7 (50 mg, 0.093 mmol) was dissolved in ultra-dry dichloromethane (5 mL), and 3-(N-phthalimido)propionic acid (40 mg, 0.182 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (80 mg, 0.449 mmol), and 4-dimethylaminopyridine (8 mg, 0.060 mmol) were added sequentially. The mixture was allowed to react overnight. LC-MS monitoring indicated the reaction was complete. The reaction solution was concentrated and purified by PTLC to give compound 4-1 (14 mg, 20.5%). MS (ESI): m / z found [M+H] + =737.2.

[0303] Synthesis of compound 4:

[0304] Under an ice-water bath, 4-1 (14 mg, 0.019 mmol) was dissolved in ultra-dry dichloromethane (5 mL) and trifluoroacetic acid (0.5 mL) was added for one hour. LC-MS monitored the reaction for completion. The reaction solution was diluted with ethyl acetate, extracted with saturated sodium bicarbonate solution, washed with saturated aqueous NaCl solution, dried over anhydrous Na2SO4, and concentrated. Pre-HPLC purification afforded compound 4 (1.9 mg, 16%). MS (ESI): m / z found [M+H] + =637.2.

[0305] Example 5:

[0306] Synthesis route:

[0307] Synthesis of intermediate 5-2:

[0308] Sodium chloroacetate (928 mg, 8.0 mmol) and sodium hydroxide (319 mg, 8.0 mmol) were dissolved in water (5 mL) at room temperature. After mixing, 3,4-methylenedioxyphenol (1.0 g, 7.2 mmol) was added and allowed to react for ten minutes. The temperature was then raised to 60°C and allowed to react overnight. LCMS monitoring indicated the reaction was complete. Hydrochloric acid was added to adjust the pH to 2. The precipitated solid was filtered, recrystallized from petroleum ether and ethyl acetate, and concentrated to yield compound 5-2 (485 mg, 34%). MS (ESI): m / z found [M+H] + =197.0.

[0309] Synthesis of intermediate 5-3:

[0310] At room temperature, 1-7 (50 mg, 0.093 mmol) was dissolved in ultra-dry dichloromethane (5 mL), and 5-2 (36 mg, 0.19 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (80 mg, 0.45 mmol), and 4-dimethylaminopyridine (8 mg, 0.06 mmol) were added sequentially. The reaction was allowed to react overnight. LC-MS monitoring showed that the reaction was complete. The reaction solution was concentrated and purified by PTLC to give compound 5-3 (53 mg, 80%). MS (ESI): m / z found [M+H] + =714.2.

[0311] Synthesis of compound 5:

[0312] Under ice-water bath, 5-3 (53 mg, 0.08 mmol) was dissolved in ultra-dry dichloromethane (5 mL) and trifluoroacetic acid (0.5 mL) was added to react for one hour. LCMS monitored the reaction to be complete. The reaction solution was dried and concentrated, and then purified by PTLC to give compound 5 (34 mg, 72%). MS (ESI): m / z found [M+H] + =614.2.

[0313] Example 6:

[0314] Synthesis route:

[0315] Synthesis of intermediate 6-2:

[0316] At room temperature, potassium hydroxide (1.8 g, 30.8 mmol) was dissolved in water (20 mL), and 5-fluorouracil (6-1, 1.0 g, 7.7 mmol) was added. The mixture was reacted at 60°C for 1 hour. The temperature was then cooled to room temperature, bromoacetic acid (1.5 g, 10.8 mmol) was added, and the temperature was raised to 60°C for 5 hours. After cooling to room temperature, hydrochloric acid was added to adjust the pH to 2, and the mixture was stirred at room temperature overnight. The precipitated solid was filtered and concentrated to obtain compound 6-2 (710 mg, 49%). MS (ESI): m / z found [M+H] + =189.0.

[0317] Synthesis of intermediate 6-3:

[0318] At room temperature, 1-7 (50 mg, 0.09 mmol) was dissolved in ultra-dry dichloromethane (5 mL), and 6-2 (35 mg, 0.19 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (80 mg, 0.45 mmol), and 4-dimethylaminopyridine (8 mg, 0.06 mmol) were added sequentially. The reaction was allowed to react overnight. LCMS monitoring showed that the reaction was complete. The reaction solution was concentrated and purified by PTLC to give compound 6-3 (50 mg, 83%). MS (ESI): m / z found [M+H] + =706.2.

[0319] Synthesis of compound 6:

[0320] Under ice-water bath, 6-3 (50 mg, 0.07 mmol) was dissolved in ultra-dry dichloromethane (5 mL) and trifluoroacetic acid (0.5 mL) was added to react for 1 hour. LCMS monitored the reaction to be complete. The reaction solution was dried and concentrated and then purified by Pre-HPLC to give compound 6 (34 mg, 70%). MS (ESI): m / z found [M+H] + =606.2.

[0321] Example 7:

[0322] Synthesis route:

[0323] Synthesis of intermediate 7-2:

[0324] At room temperature, 1-7 (50 mg, 0.09 mmol) was dissolved in 5 mL of ultra-dry dichloromethane. 4,7,10,13-Tetraoxatetradecanoic acid (43 mg, 0.182 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (80 mg, 0.449 mmol), and 4-dimethylaminopyridine (7.4 mg, 0.060 mmol) were added sequentially and allowed to react overnight. LC-MS monitoring indicated the reaction was complete. The reaction solution was concentrated and purified by PTLC to give compound 7-2 (48 mg, 68%). MS (ESI): m / z found [M+H] + =637.2.

[0325] Synthesis of compound 7:

[0326] Under ice-water bath, 7-2 (48 mg, 0.064 mmol) was dissolved in ultra-dry dichloromethane (5 mL) and trifluoroacetic acid (1.0 mL) was added to react for one hour. LC-MS monitored the reaction to be complete. The reaction solution was dried and concentrated, and then purified by Pre-HPLC to give compound 7 (35.87 mg, 86%). MS (ESI): m / z found [M+H] + =654.0.

[0327] Example 8:

[0328] Synthesis route:

[0329] Synthesis of Example 8:

[0330] Glycolic acid (19 mg, 0.248 mmol) was dissolved in DMF (3 mL), and HOSU (28.5 mg, 0.248 mmol) and EDCI (48 mg, 0.248 mmol) were added. After reacting at room temperature for 1 hour, compound 6 (100 mg, 0.165 mmol) and TEA (18.4 mg, 0.182 mmol) were added and reacted at room temperature overnight. After the reaction was complete as monitored by LC-MS, the reaction solution was purified by pre-HPLC and lyophilized to obtain a white solid product (28.4 mg, 26.0%). MS (ESI): m / z found [M+H] + =664.0.

[0331] Example 9:

[0332] Synthesis route:

[0333] Synthesis of intermediate 9-2:

[0334] Compound 1-7 (30 mg, 0.056 mmol) was dissolved in DCM (3 mL), and 2-methyl-4-chlorophenoxyacetic acid (20 mg, 0.098 mmol), EDCI (33 mg, 0.169 mmol), and DMAP (4.1 mg, 0.034 mmol) were added. The mixture was allowed to react at room temperature overnight. After the reaction was complete as monitored by LC-MS, the mixture was extracted with water / DCM, concentrated by distillation under reduced pressure, and purified by TLC (DCM:MeOH=10:1) to give 9-2 (31 mg, 77.2%) as a white solid. MS (ESI): m / z found [M+H] + =718.0.

[0335] Synthesis of compound 9:

[0336] Compound 9-2 (30 mg, 0.042 mmol) was dissolved in DCM (3 mL) and TFA (0.3 mL) was added. The mixture was allowed to react at room temperature for 3 hours. After the reaction was complete as monitored by LC-MS, the mixture was evaporated to dryness under reduced pressure, prepared by HPLC, and lyophilized to obtain a white solid product (11.9 mg, 45.9%). MS (ESI): m / z found [M+H] + =618.0.

[0337] Example 10:

[0338] Synthesis route:

[0339] Synthesis of intermediate 10-3:

[0340] Podophyllotoxin (10-1, 100 mg, 0.241 mmol) was dissolved in 10 mL of DCM at room temperature. TEA (49 mg, 0.482 mmol), succinic anhydride (25 mg, 0.241 mmol), and DMAP (3.0 mg, 0.024 mmol) were added sequentially and allowed to react at room temperature for 4 hours. TLC monitoring indicated the reaction was complete. The reaction solution was concentrated to dryness and separated by column chromatography (dichloromethane:methanol = 10:1) to afford compound 10-3 (125 mg, 99% yield). MS (ESI): m / z found [MH] - =513.0.

[0341] Synthesis of intermediate 10-4:

[0342] Compound 10-3 (50 mg, 0.093 mmol) was dissolved in dry DCM, and 1-7 (94 mg, 0.182 mmol) was added sequentially. The mixture was allowed to react overnight at room temperature. TLC monitoring showed that the reaction of the starting materials was complete. The reaction solution was concentrated to dryness and then separated by column chromatography (dichloromethane:methanol=10:1) to obtain compound 10-4 (37 mg, yield 39%). MS (ESI): m / z found [M+H] + =1032.2.

[0343] Synthesis of compound 10:

[0344] At room temperature, 10-4 (10 mg, 0.0097 mmol) was dissolved in 5 mL of dry DCM, and zinc bromide (4.4 mg, 0.0194 mmol) was added. The reaction was allowed to react at room temperature for 15 hours. LC-MS monitoring showed that the reaction was complete. The reaction solution was concentrated to dryness and then dissolved in a small amount of DMF and purified by pre-HPLC to obtain compound 10 (2.86 mg, 32% yield). MS (ESI): m / z found [M+H] + =932.4.

[0345] Example 11:

[0346] Synthesis route:

[0347] Synthesis of intermediate 11-2:

[0348] Boron trichloride (32 mL, 31.96 mmol) was dissolved in 80 mL of DCM under an ice-water bath. 3-Fluoro-4-methylaniline (11-1, 5 g, 39.95 mmol), chloroacetonitrile (3.53 g, 46.95 mmol), and aluminum trichloride (6.9 g, 51.94 mmol) were added sequentially. The mixture was stirred at 0°C for 10 minutes, then warmed to room temperature and allowed to react for 10 minutes before being heated again to 40°C overnight. TLC monitoring indicated the reaction was complete. The reaction solution was cooled to ice water, 30 mL of water was added, and the mixture was stirred in an ice bath for 10 minutes. 2N hydrochloric acid solution was added and stirred for one hour. The mixture was diluted with dichloromethane, washed three times with water, and the combined organic phases were dried over anhydrous Na2SO4, concentrated, and separated by column chromatography (dichloromethane:methanol = 10:1) to yield compound 11-2 (2.15 g, 27%). MS (ESI): m / z found [M+H] + =202.0.

[0349] Synthesis of intermediate 11-3:

[0350] At room temperature, compound 11-2 (500 mg, 2.487 mmol) was dissolved in 5 mL of toluene, and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-F]indolizine-3,6,10(4H)-one (624 mg, 2.369 mmol) and PPTS (30 mg, 0.118 mmol) were added. The atmosphere was purged with nitrogen three times, and the temperature was raised to 110°C for overnight reaction. LC-MS monitoring showed that the starting material had reacted completely. The reaction solution was concentrated to dryness and then dissolved in a small amount of DMF and separated by reverse phase column chromatography (water:acetonitrile = 45:55) to obtain compound 11-3 (467 mg, 46% yield). MS (ESI): m / z found [M+H] + =429.0.

[0351] Synthesis of intermediate 11-4:

[0352] Compound 11-3 (754 mg, 1.758 mmol) was dissolved in 6 mL of dry DMSO at room temperature, and sodium azide (114 mg, 1.758 mmol) was added. The mixture was allowed to react at room temperature for 4 hours. LC-MS monitoring showed that the reaction of the starting material was complete. The reaction solution was placed in an ice bath, 50 mL of water was added, and the mixture was stirred for 10 minutes before being filtered. The filter cake was collected and dried to obtain compound 11-4 (603 mg, 78% yield). MS (ESI): m / z found [M+H] + =436.0.

[0353] Synthesis of intermediate 11-5:

[0354] At room temperature, compound 11-4 (600 mg, 1.378 mmol) was dissolved in 10 mL of toluene, and triethyl phosphite (590 μL, 3.445 mmol) was added. The temperature was raised to 110°C and the reaction was allowed to react for 4 hours. After cooling to room temperature, 5 mL of 3N methanolic hydrogen chloride solution was added, and the temperature was raised to 80°C again, and the reaction was maintained overnight. LC-MS monitoring showed that the starting material had reacted completely. The reaction solution was concentrated and purified by reverse-phase column chromatography to obtain compound 11-5 (360 mg, 63% yield). MS (ESI): m / z found [M+H] + =410.0.

[0355] Synthesis of intermediate 11-6:

[0356] Compound 11-5 (64 mg, 0.156 mmol) was dissolved in 5 mL of dry pyridine at room temperature, and TESOTf (340 μL, 1.56 mmol) was added and allowed to react overnight. The reaction was monitored for completion by TLC. FmocCl (81 mg, 0.312 mmol) was added and allowed to react for two hours, which was also monitored for completion by TLC. The reaction solution was dried, diluted with DCM, and washed three times with water and once with saturated aqueous NaCl. After drying and concentration over anhydrous NaSO, the product was purified by column chromatography (dichloromethane:methanol = 95:5) to afford 11-6 (105 mg, 90% yield).

[0357] Synthesis of intermediate 11-7:

[0358] At room temperature, compound 11-6 (105 mg, 0.141 mmol) was dissolved in 5 mL of toluene, and Lawesson's reagent (143 mg, 0.352 mmol) was added. The temperature was raised to 110 ° C. and the reaction was complete after TLC monitoring. After the reaction solution was concentrated, a small amount of dichloromethane was added for dissolution and purified by column chromatography (dichloromethane: methanol = 95:5) to obtain compound 11-7 (102 mg, yield 95%).

[0359] Synthesis of intermediate 11-8:

[0360] Compound 11-7 (102 mg, 0.134 mmol) was dissolved in 3 mL of dry DMF at room temperature, and morpholine (300 μL) was added and allowed to react for 4 hours. The reaction was complete as monitored by TLC. The reaction solution was purified by reverse phase column chromatography (water:acetonitrile = 45:55) to obtain compound 11-8 (47 mg, 65% yield). MS (ESI): m / z found [M+H] + =540.2.

[0361] Synthesis of compound 11:

[0362] Compound 11-8 (47 mg, 0.0871 mmol) was dissolved in 5 mL of dry DCM at room temperature, and trifluoroacetic acid (1 mL) was added. The reaction was allowed to react overnight at room temperature. LC-MS monitored the reaction to completion. The reaction solution was purified by reverse phase column chromatography (water:acetonitrile = 45:55) to obtain compound 11 (32 mg, 86% yield). MS (ESI): m / z found [M+H] + =426.0.

[0363] Example 12:

[0364] Synthesis route:

[0365] Synthesis of intermediate 12-2:

[0366] DCM (50 ml) was placed in an eggplant-shaped flask and cooled on ice. Boron trichloride (11.34 ml, 11.34 mmol) was added and ice-cooled for 10 minutes. 3-Fluoro-4-methoxyaniline (12-1, 2 g, 14.17 mmol) was then added. Chloroacetonitrile (1.05 ml, 16.65 mmol) was then added after an ice-cooling for 10 minutes. Aluminum trichloride (2.46 g, 18.42 mmol) was then added after an ice-cooling for 10 minutes. The mixture was then heated to 40°C overnight. After completion of the reaction as monitored by LCMS, 15 ml of water was added under ice-cooling and stirred for 10 minutes. 4 ml of 2M hydrochloric acid was added and the mixture was stirred at room temperature for 1.5 hours. The mixture was filtered and the filtrate was extracted with water / DCM. Purification by normal phase column chromatography (PE:EA=2:1) ​​afforded the product as a yellow solid (668.8 mg, 21.7%). MS (ESI): m / z found [M+H] + =218.15

[0367] Synthesis of intermediate 12-3:

[0368] Compound 12-2 (668.8 mg, 3.07 mmol) was dissolved in toluene (15 ml), and 7-ethyl-10-hydroxycamptothecin intermediate (808.2 mg, 3.07 mmol) and PPTS (38.7 mg, 0.154 mmol) were added. The mixture was allowed to react overnight at 110°C. After completion of the reaction as monitored by LC-MS, the mixture was filtered, the supernatant was washed with ethanol, and dried to obtain a light yellow solid product (974.4 mg, 71.5%). MS (ESI): m / z found [M+H] + =445.25.

[0369] Synthesis of intermediate 12-4:

[0370] Compound 12-3 (974.4 mg, 2.19 mmol) was dissolved in DMSO (15 ml), sodium azide (142.4 mg, 2.19 mmol) was added, and the mixture was allowed to react at room temperature for 3 hours. After the reaction was complete as monitored by LCMS, 8 ml of water was added, the mixture was filtered, and dried to obtain a brown solid product (801.4 mg). MS (ESI): m / z found [M+H] + =452.30.

[0371] Synthesis of intermediate 12-5:

[0372] Compound 12-4 (801.4 mg, 1.78 mmol) was dissolved in ethanol (10 ml), and triethyl phosphite (739.54 mg, 4.45 mmol) was added. The mixture was reacted at 50°C for 4 hours, cooled to room temperature, and a solution of hydrogen chloride in ethyl acetate (19 ml) was added. The mixture was reacted at 60°C overnight. After completion of the reaction as monitored by LCMS, the mixture was filtered to obtain a light yellow solid product (550.8 mg, 72.8%). MS (ESI): m / z found [M+H] + =426.30.

[0373] Synthesis of intermediate 12-6:

[0374] Compound 12-5 (550.8 mg, 1.29 mmol) was dissolved in pyridine (10 ml), and TESOTf (2.8 ml, 12.9 mmol) was added. The mixture was allowed to react at room temperature overnight. After completion of the reaction as monitored by LCMS, FmocCl (667.4 mg, 2.58 mmol) was added and allowed to react at room temperature for 3 hours. After completion of the reaction as monitored by TLC, the pyridine was dried, and the mixture was extracted with H2O / DCM. The product was purified by normal phase column chromatography to obtain the product as a yellow solid (467.5 mg, 47.6%).

[0375] Synthesis of intermediate 12-7:

[0376] Under nitrogen, compound 12-6 (467.5 mg, 0.61 mmol) was dissolved in toluene (10 ml), and Lawesson's reagent (744.1 mg, 1.84 mmol) was added. The mixture was allowed to react at 110°C for 4 hours. After completion of the reaction, the mixture was evaporated to dryness under reduced pressure and purified by normal phase column chromatography to obtain a yellow solid (389.3 mg, 82.1%).

[0377] Synthesis of intermediate 12-8:

[0378] Compound 12-7 (389.3 mg, 0.5 mmol) was dissolved in DMF (5 ml), and morpholine (2 ml) was added. The mixture was allowed to react at room temperature for 5 hours. After completion of the reaction as monitored by LCMS, the mixture was concentrated by vacuum distillation, extracted with saturated NaCl solution / DCM, and purified by normal column chromatography (DCM:MeOH = 47:3). A brown solid product (134.6 mg, 48.4%) was obtained. MS (ESI): m / z found [M+H] + =556.45.

[0379] Synthesis of compound 12:

[0380] Compound 12-8 (134.6 mg, 0.24 mmol) was dissolved in DCM (5 ml) and TFA (1 ml) was added. The mixture was allowed to react at room temperature overnight. After completion of the reaction as monitored by LCMS, the reaction solution was dried and purified by reverse-phase column chromatography (H2O:ACN = 4:1). Lyophilization afforded an orange-red solid (54.9 mg, 51.8%). MS (ESI): m / z found [M+1] + =442.30. 1 H NMR (400 MHz, DMSO-d6) δ 8.15 (s, 1H), 8.04 (d, J = 12.1 Hz, 1H), 7.79 (d, J = 8.6 Hz, 2H), 6.70 (s, 1H), 5.93 (d, J = 16.6 Hz, 1H), 5.76 (s, 2H), 5.53 (d, J = 16.6 Hz, 1H), 4.48 (s, 2H), 4.11 (s, 3H), 1.90 (dt, J = 10.5, 6.8 Hz, 2H), 0.86 (t, J = 7.3 Hz, 3H). Example 13:

[0381] Synthesis route:

[0382] Synthesis of intermediate 13-2:

[0383] Boron trichloride (26 ml, 26 mmol) was dissolved in 80 ml of dichloromethane under an ice-water bath. 6-amino-1,4-benzodioxetine (13-1, 5 g, 33 mmol), chloroacetonitrile (3.01 g, 40 mmol), and aluminum trichloride (5.7 g, 43 mmol) were added sequentially. The mixture was stirred at 0°C for 10 minutes, then warmed to room temperature and reacted for 10 minutes before being heated again to 40°C and held overnight. TLC monitoring indicated the reaction was complete. The reaction solution was cooled to ice water, and 30 ml of water was added and stirred in an ice-water bath for 10 minutes. 2N hydrochloric acid was added and stirred for one hour. The mixture was diluted with dichloromethane and washed three times with water. The combined organic phases were dried over anhydrous Na2SO4, concentrated, and separated by column chromatography (dichloromethane:methanol = 10:1) to afford compound 13-2 (1.8 g, 24%). MS (ESI): m / z found [M+H] + =228.00.

[0384] Synthesis of intermediate 13-3:

[0385] At room temperature, compound 13-2 (2.6 g, 11.42 mmol) was dissolved in 60 mL of toluene, and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-F]indolizine-3,6,10(4H)-one (3.0 g, 11.42 mmol) and PPTS (144 mg, 0.57 mmol) were added. After nitrogen substitution three times, the temperature was raised to 110°C and the reaction was allowed to proceed overnight. LC-MS monitoring showed that the starting material had reacted completely. After cooling the reaction solution, the product was slurried and washed with ethanol to obtain compound 13-3 (2.74 g, 54% yield). MS (ESI): m / z found [M+H] + =455.30.

[0386] Synthesis of intermediate 13-4:

[0387] Compound 13-3 (2.74 g, 6.02 mmol) was dissolved in 6 ml of dry DMSO at room temperature, and sodium azide (392 mg, 6.02 mmol) was added. The mixture was allowed to react at room temperature for 4 hours. LC-MS monitoring showed that the reaction of the starting material was complete. The reaction solution was placed in an ice bath, 50 ml of water was added, and the mixture was stirred for 10 minutes before being filtered. The filter cake was collected and dried to obtain compound 13-4 (2.32 g, 83%). MS (ESI): m / z found [M+H] + =462.30.

[0388] Synthesis of intermediate 13-5:

[0389] At room temperature, compound 13-4 (2.32 g, 5.019 mmol) was dissolved in 20 ml of ethanol, and triethyl phosphite (2.15 ml, 12.548 mmol) was added. The reaction was heated to 50°C for 4 hours. After cooling to room temperature, 30 ml of 3N methanolic hydrogen chloride solution was added, and the temperature was raised to 60°C again, and the reaction was maintained overnight. LC-MS monitoring showed that the starting material had reacted completely. The reaction solution was cooled to room temperature, and the precipitated solid was filtered and washed with ethanol to obtain compound 13-5 (1.657 g, 73% yield). MS (ESI): m / z found [M+H] + =436.0.

[0390] Synthesis of intermediate 13-6:

[0391] Compound 13-5 (1.657 g, 3.805 mmol) was dissolved in 15 ml of dry pyridine at room temperature, and triethylsilyl trifluoromethanesulfonate (8.2 ml, 38.05 mmol) was added and allowed to react overnight. TLC monitored the reaction to be complete. Then, 9-fluorenylmethyl chloroformate (1.97 g, 7.610 mmol) was added and allowed to react for two hours. TLC monitored the reaction to be complete. The reaction solution was dried, diluted with dichloromethane, and washed three times with water and once with saturated aqueous NaCl. After drying and concentration over anhydrous NaSO, the solution was purified by column chromatography (dichloromethane:methanol = 95:5) to afford 36-6 (1.58 g, 58% yield).

[0392] Synthesis of intermediate 13-7:

[0393] At room temperature, compound 36-6 (1.58 g, 2.047 mmol) was dissolved in 5 ml of toluene, and Lawesson's reagent (2.5 g, 6.140 mmol) was added. The temperature was raised to 100°C and the reaction was allowed to proceed for 4 hours. The reaction was monitored to be complete by TLC. The reaction solution was concentrated and dissolved in a small amount of dichloromethane and purified by column chromatography (dichloromethane: methanol = 95:5) to obtain compound 13-7 (499 mg, yield 31%).

[0394] Synthesis of intermediate 13-8:

[0395] Compound 13-7 (499 mg, 0.633 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide at room temperature, and morpholine (2 ml) was added and allowed to react for 4 hours. TLC confirmed the completion of the reaction. The reaction solution was diluted with dichloromethane, washed three times with water, and once with a saturated aqueous NaCl solution. After drying over anhydrous Na2SO4, it was concentrated and purified by normal phase column chromatography (dichloromethane:methanol = 95:5) to obtain compound 13-8 (317 mg, 88% yield). MS (ESI): m / z found [M+H] + =566.20.

[0396] Synthesis of compound 13:

[0397] Compound 13-8 (257 mg, 0.453 mmol) was dissolved in 5 ml of dry dichloromethane at room temperature, and trifluoroacetic acid (2 ml) was added. The reaction was allowed to react overnight at room temperature. LC-MS monitored the reaction to completion. The reaction solution was purified by reverse phase column chromatography (water:acetonitrile = 45:55) to obtain compound 13 (112 mg, 55% yield). MS (ESI): m / z found [M+H] + =452.35. 1H NMR (600MHz, DMSO-d6) δ8.32(s,2H),7.92(s,1H),7.79(s,1H),7.71(s,1H),6.71(s,1H),5.94(d,J=16.5Hz,1H),5.77(d,J=2. 0Hz,2H),5.54(d,J=16.5Hz,1H),4.69(d,J=5.9Hz,2H),4.54–4.46(m,4H),1.97–1.85(m,J=7.2Hz,2H),0.87(t,J=7.3Hz,3H).

[0398] Example 14:

[0399] Synthesis route:

[0400] Synthesis of intermediate 14-2:

[0401] DCE (300 ml) was placed in an eggplant flask and cooled on ice. Boron trichloride (56.5 ml, 56.5 mmol) was added. After 10 minutes of ice-bath, 3-chloro-p-toluidine (14-1) (10 g, 70.6 mmol) was added. After 10 minutes of ice-bath, chloroacetonitrile (5.25 ml, 82.96 mmol) was added. After 10 minutes of ice-bath, aluminum trichloride (12.24 g, 91.78 mmol) was added. The mixture was stirred at room temperature for 10 minutes and then heated to 80°C overnight. After completion of the reaction as monitored by LCMS, 80 ml of water was added under ice-bath, and the mixture was stirred for 10 minutes. 20 ml of 2M hydrochloric acid was added, and the mixture was stirred at room temperature for 1.5 hours. The mixture was filtered, and the filtrate was extracted with water / DCM and purified by normal phase column chromatography (PE:EA=2:1) ​​to give the product as a yellow solid (2.96 g, 19.2%). MS (ESI): m / z found [M+1] + =218.0.

[0402] Synthesis of intermediate 14-3:

[0403] Compound 14-2 (2.96 g, 13.57 mmol) was dissolved in toluene (65 ml), and 7-ethyl-10-hydroxycamptothecin intermediate (3.57 g, 13.57 mmol) and PPTS (170.6 mg, 0.679 mmol) were added. The mixture was allowed to react overnight at 110°C. After completion of the reaction as monitored by LCMS, the mixture was filtered, the supernatant was washed with ethanol, and dried to obtain a pale yellow solid product (5 g, 82.8%). MS (ESI): m / z found [M+1]+ = 445.0.

[0404] Synthesis of intermediate 14-4:

[0405] Compound 14-3 (5 g, 11.22 mmol) was dissolved in DMSO (80 ml), sodium azide (729.4 mg, 11.22 mmol) was added, and the mixture was allowed to react at room temperature for 3 hours. After the reaction was complete as monitored by LCMS, 80 ml of water was added, the mixture was filtered, and dried to obtain a brown solid product (4.88 g). MS (ESI): m / z found [M+H] + =452.0.

[0406] Synthesis of intermediate 14-5:

[0407] Compound 14-4 (4.88 g, 10.8 mmol) was dissolved in ethanol (50 ml), and triethyl phosphite (4.63 ml, 27 mmol) was added. The mixture was reacted at 50°C for 4 hours, then cooled to room temperature. A solution of hydrogen chloride in ethyl acetate (60 ml) was added, and the mixture was allowed to react at 60°C overnight. After completion of the reaction as monitored by LCMS, the mixture was filtered to obtain a pale yellow solid product (4.39 g, 95.4%). MS (ESI): m / z found [M+H] + =426.0.

[0408] Synthesis of intermediate 14-6:

[0409] Compound 14-5 (4.39 g, 10.31 mmol) was dissolved in pyridine (40 ml), and TESOTf (22.19 ml, 103.1 mmol) was added. The mixture was allowed to react at room temperature overnight. After completion of the reaction as monitored by LCMS, FmocCl (5.34 g, 20.62 mmol) was added and allowed to react at room temperature for 3 hours. After completion of the reaction as monitored by TLC, the pyridine was evaporated, and the mixture was extracted with H2O / DCM. The product was purified by normal phase column chromatography to obtain the product as a yellow solid (2.94 g, 97%).

[0410] Synthesis of intermediate 14-7:

[0411] Under nitrogen, compound 14-6 (2.94 g, 3.86 mmol) was dissolved in toluene (30 ml), and Lawesson's reagent (4.68 g, 11.58 mmol) was added. The mixture was allowed to react at 110°C for 4 hours. After completion of the reaction, the mixture was evaporated to dryness under reduced pressure and purified by normal phase column chromatography to obtain a yellow solid (1.51 g, 50.3%).

[0412] Synthesis of intermediate 14-8:

[0413] Compound 14-7 (1.5 g, 1.97 mmol) was dissolved in DMF (5 ml), and morpholine (2 ml) was added. The mixture was allowed to react at room temperature for 5 hours. After completion of the reaction as monitored by LCMS, the mixture was concentrated by vacuum distillation, extracted with saturated NaCl solution / DCM, and purified by normal column chromatography (DCM:MeOH = 20:1). A brown solid product (536.8 mg, 49%) was obtained. MS (ESI): m / z found [M+H] + =556.2.

[0414] Synthesis of compound 14:

[0415] Compound 14-8 (536.8 mg, 0.97 mmol) was dissolved in DCM (10 ml) and TFA (2 ml) was added. The mixture was allowed to react at room temperature overnight. After completion of the reaction as monitored by LCMS, the reaction mixture was dried and purified by reverse-phase column chromatography (H2O:ACN=3:1). Lyophilization afforded an orange-red solid (348.2 mg, 81.2%). MS (ESI): m / z found [M+H] + =442.30. 1 H NMR (400MHz, DMSO-d6) δ8.40(s,1H),8.31(d,J=2.6Hz,1H),8.15(s,2H),7.81(s,1H),6.73(s,1H),5.94(d,J=16.6Hz, 1H), 5.77 (s, 2H), 5.54 (d, J = 16.7Hz, 1H), 4.52 (s, 1H), 2.61 (s, 3H), 1.90 (dt, J = 9.8, 6.7Hz, 2H), 0.87 (t, J = 7.3Hz, 4H).

[0416] Example 15:

[0417] Synthesis route:

[0418] Synthesis of intermediate 15-2:

[0419] DCE (300 ml) was placed in an eggplant-shaped flask and cooled on ice. Boron trichloride (61.6 ml, 61.6 mmol) was added. After 10 minutes of ice-cooling, 3,4-difluoroaniline (57-a) (10 g, 77.46 mmol) was added. After 10 minutes of ice-cooling, chloroacetonitrile (5.76 ml, 91.02 mmol) was added. After 10 minutes of ice-cooling, aluminum trichloride (13.4 g, 100.7 mmol) was added. The mixture was stirred at room temperature for 10 minutes and then heated to 80°C overnight. After completion of the reaction as monitored by LCMS, 80 ml of water was added under ice-cooling and stirred for 10 minutes. 20 ml of 2M hydrochloric acid was added and the mixture was stirred at room temperature for 1.5 hours. The mixture was filtered and the filtrate was extracted with water / DCM. Purification by normal phase column chromatography (PE:EA=2:1) ​​afforded the product as a yellow solid (5.79 g, 36.4%). MS (ESI): m / z found [M+H] + =206.0

[0420] Synthesis of intermediate 15-3:

[0421] Compound 15-2 (5.79 g, 28.16 mmol) was dissolved in toluene (150 ml), and 7-ethyl-10-hydroxycamptothecin intermediate (7.4 g, 28.16 mmol) and PPTS (354.3 mg, 1.41 mmol) were added. The mixture was allowed to react overnight at 110°C. After completion of the reaction as monitored by LC-MS, the mixture was filtered, the supernatant solid was washed with ethanol, and dried to yield the product as a pale yellow solid (6.87 g, 56.4%). MS (ESI): m / z found [M+H]+ = 433.0.

[0422] Synthesis of intermediate 15-4:

[0423] Compound 15-3 (6.87 g, 15.87 mmol) was dissolved in DMSO (105 ml), sodium azide (1.03 mg, 15.87 mmol) was added, and the mixture was allowed to react at room temperature for 3 hours. After the reaction was complete as monitored by LCMS, 80 ml of water was added, the mixture was filtered, and dried to obtain a brown solid product (5.45 g). MS (ESI): m / z found [M+H] + =440.0.

[0424] Synthesis of intermediate 15-5:

[0425] Compound 15-4 (5.45 g, 12.4 mmol) was dissolved in ethanol (75 ml), and triethyl phosphite (5.35 ml, 31.03 mmol) was added. The mixture was reacted at 50°C for 4 hours, then cooled to room temperature. A solution of hydrogen chloride in ethyl acetate (120 ml) was added, and the mixture was reacted at 60°C overnight. After completion of the reaction as monitored by LCMS, the mixture was filtered to obtain a pale yellow solid product (3.39 g, 51.67%). MS (ESI): m / z found [M+H] + =414.0.

[0426] Synthesis of intermediate 15-6:

[0427] Compound 15-5 (3.39 g, 8.2 mmol) was dissolved in pyridine (33 ml), and TESOTf (17.65 ml, 82 mmol) was added. The mixture was allowed to react at room temperature overnight. After completion of the reaction as monitored by LCMS, FmocCl (4.24 g, 16.4 mmol) was added and the reaction was continued at room temperature for 3 hours. After completion of the reaction as monitored by TLC, the pyridine was dried, and the mixture was extracted with H2O / DCM. The product was purified by normal phase column chromatography to obtain the product as a yellow solid (2.59 g, 42.1%).

[0428] Synthesis of intermediate 15-7:

[0429] Under nitrogen, compound 15-6 (2.59 g, 3.45 mmol) was dissolved in toluene (65 ml), and Lawesson's reagent (4.19 g, 10.35 mmol) was added. The mixture was allowed to react at 110°C for 4 hours. After completion of the reaction, the mixture was evaporated to dryness under reduced pressure and purified by normal phase column chromatography to obtain a yellow solid (1.69 g, 64%).

[0430] Synthesis of intermediate 15-8:

[0431] Compound 15-7 (1.69 g, 2.21 mmol) was dissolved in DMF (10 ml), and morpholine (4 ml) was added. The mixture was allowed to react at room temperature for 5 hours. After completion of the reaction as monitored by LCMS, the mixture was concentrated by vacuum distillation, extracted with saturated NaCl solution / DCM, and purified by normal column chromatography (DCM:MeOH = 25:2). A brown solid product (486.8 mg, 40.6%) was obtained. MS (ESI): m / z found [M+H] + =544.4.

[0432] Synthesis of compound 15:

[0433] Compound 15-8 (486.8 mg, 0.89 mmol) was dissolved in DCM (10 ml) and TFA (2 ml) was added. The mixture was allowed to react at room temperature overnight. After completion of the reaction as monitored by LCMS, the reaction solution was dried and purified by reverse-phase column chromatography (H2O:ACN=5:1). Lyophilization afforded an orange-red solid (128.8 mg, 51.8%). MS (ESI): m / z found [M+H] + =430.0. 1 H NMR (600MHz, DMSO-d6) δ8.53(dd,J=12.2,8.5Hz,1H),8.33(dd,J=11.4,7.9Hz,1H),7.84(s,1H),6.73(s,1H),5.94(d,J=1 6.5Hz,1H),5.81(d,J=2.0Hz,2H),5.55(d,J=16.5Hz,1H),4.56(s,2H),1.95–1.86(m,J=7.3Hz,2H),0.87(t,J=7.3Hz,3H).

[0434] Example 16:

[0435] Synthesis route:

[0436] Synthesis of intermediate 16-2:

[0437] Under ice, 16-1 (10 g, 0.045 mol) was dissolved in DCM (100 mL). CDI (9.5 g, 0.058 mol) was added and stirred for one hour. Dimethylhydroxylamine hydrochloride (6.5 g, 0.067 mol) and triethylamine (13.6 g, 0.013 mol) were then added. The mixture was returned to room temperature and stirred overnight. TLC confirmed the complete reaction. The mixture was quenched with 50 mL of water and extracted three times with dichloromethane. The organic phases were combined and washed once with 1N hydrochloric acid, once with saturated sodium bicarbonate, and once with saturated brine. The mixture was dried over anhydrous sodium sulfate, filtered, and dried to afford 16-2 (11 g, 92%) as a white solid. LC-MS (ESI): m / z found [M+H] + =267.2.

[0438] Synthesis of intermediate 16-3;

[0439] Under an ice-water bath, 3-fluoro-4-methylaniline (7.8 g, 0.062 mol) was dissolved in dichloromethane (50 mL). Acetic anhydride (12.7 g, 0.124 mol) and triethylamine (20.2 g, 0.2 mol) were added and the mixture was allowed to react at room temperature for 3 hours. TLC confirmed the complete reaction. The mixture was quenched with water and extracted with dichloromethane. The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and slurried with petroleum ether to obtain a brown solid 16-3 (11 g, 94.7%). MS (ESI): m / z found [M+H] + =168.0.

[0440] Synthesis of intermediate 16-4:

[0441] Under an ice-water bath, 16-3 (11 g, 0.065 mol) was added to DMF (110 mL), followed by the addition of NBS (16.3 g, 0.092 mol) in portions. The mixture was stirred at room temperature overnight. TLC confirmed the complete reaction. The mixture was poured into ice water and stirred for 0.5 h. The mixture was filtered, and the filter cake was washed with saturated sodium bicarbonate and dried to afford 16-4 (12 g, 74.5%) as a white solid. MS (ESI): m / z found [M+H] + =246.0.

[0442] Synthesis of intermediate 16-5:

[0443] Compound 16-4 (2 g, 8.13 mmol) was added to THF, the atmosphere was replaced with nitrogen, and the system was cooled to -60°C. n-Butyl lithium (7.15 mL, 17.9 mmol) was added dropwise to the system and stirred at this temperature for 1 hour. 16-2 (3.2 g, 12.19 mmol) was then dissolved in THF and added dropwise to the system at this temperature. The system was cooled to -20°C and stirred for 1 hour. The reaction was complete. Saturated ammonium chloride was added to quench the reaction at -20°C, and the mixture was extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and purified by medium-pressure normal phase chromatography to obtain a yellow oil (340 mg, 11.23%). LC-MS (ESI): m / z found [M+H] + =373.2.

[0444] Synthesis of intermediate 16-6:

[0445] 16-5 (150 mg, 0.4 mmol) was added to THF (5 mL) and methanol (5 mL). 3N aqueous hydrochloric acid (5 mL) was added and the mixture was heated to 70°C and allowed to react overnight. TLC confirmed the complete reaction. The mixture was cooled to room temperature and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and dried to give a yellow oil (110 mg, 82.67%). LC-MS (ESI): m / z found [M+H] + =331.2.

[0446] Synthesis of intermediate 16-7:

[0447] At room temperature, 16-6 (97.19 mg, 0.35 mmol) was dissolved in 3 mL of toluene, and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-F]indolizine-3,6,10(4H)-one (110 mg, 0.33 mmol) and PPTS (4.4 mg, 0.02 mmol) were added. The atmosphere was purged with nitrogen three times, and the temperature was raised to 110°C for overnight reaction. LC-MS monitoring showed that the starting material had reacted completely. The system was cooled to room temperature and filtered directly. The filter cake was washed with ethanol and then dried to obtain a brown solid 16-7 (41 mg, 25%). MS (ESI): m / z found [M+H] + =558.45.

[0448] Synthesis of intermediate 16-8:

[0449] Compound 16-7 (41 mg, 0.07 mmol) was dissolved in 3 mL of dry pyridine at room temperature, and TESOTf (194.6 mg, 0.7 mmol) was added and allowed to react overnight. TLC monitored the reaction for completion. The reaction solution was dried, diluted with DCM, and washed three times with water and once with a saturated aqueous NaCl solution. After drying over anhydrous Na2SO4, the solution was concentrated and purified by column chromatography (petroleum ether:ethyl acetate = 65:35) to afford 16-8 (50 mg, 99% yield) as a black solid. MS (ESI): m / z found [M+H] + =672.0.

[0450] Synthesis of intermediate 16-9:

[0451] At room temperature, compound 16-8 (50 mg, 0.074 mmol) was dissolved in 3 mL of toluene, and Lawesson's reagent (75.25 mg, 0.186 mmol) was added. The reaction temperature was raised to 110°C and allowed to react for 4 hours. TLC monitored the reaction until complete. The reaction solution was concentrated and dissolved in a small amount of dichloromethane and purified by column chromatography (petroleum ether:ethyl acetate = 75:25) to obtain compound 16-9 as a yellow oil (30 mg, yield 58.59%). MS (ESI): m / z found [M+H] + =688.0.

[0452] Synthesis of intermediate 16-10:

[0453] Compound 16-9 (30 mg, 0.0445 mmol) was dissolved in 5 mL of dry DCM at room temperature. Trifluoroacetic acid (1 mL) was added and the mixture was allowed to react overnight at room temperature. LC-MS monitored the reaction for completion. The mixture was then spin-dried to obtain a yellow oil, 16-10 (14 mg, 54.82%), which was used directly in the next step.

[0454] Synthesis of compound 16:

[0455] 16-10 (7 mg, 0.012 mmol) was added to a solution of hydrobromic acid in acetic acid (2 mL) at room temperature and allowed to react overnight. LC-MS confirmed the complete reaction. The reaction mixture was directly dried and purified by reverse-phase column chromatography (water:acetonitrile = 65:35) to afford compound 16 (0.38 mg, 7.09% yield). 1 HNMR (600MHz, DMSO-d6) δ8.64(d,J=5.6Hz,3H),8.49(d,J=7.8Hz,1H),8.06(d,J=10.3Hz,1H),7.85(s,1H),5.93(t,J=17.2Hz,2H),5.78(dd,J=19.5, 3.5Hz,1H),5.56(dd,J=16.5,4.0Hz,1H),5.50(s,1H),2.58(s,3H),1.92( ttd,J=14.1,7.1,2.9Hz,2H),1.83(t,J=6.2Hz,3H),0.87(t,J=7.3Hz,3H).

[0456] Example 17:

[0457] Synthesis route:

[0458] Synthesis of compound 17:

[0459] At room temperature, glycolic acid (1.4 mg, 0.0176 mmol) was dissolved in 3 mL of dry DMF, and HOSU (2.1 mg, 0.0176 mmol) and EDCI (3.4 mg, 0.0176 mmol) were added and allowed to react for one hour. Compound 11 (5.0 mg, 0.0118 mmol) and triethylamine (1.4 mg, 0.0130 mmol) were then added and allowed to react overnight at room temperature. LC-MS monitored the reaction completion, and the reaction solution was purified by pre-HPLC to yield compound 17 (3.49 mg, 61% yield). MS (ESI): m / z found [M+H] + =484.0.

[0460] Example 18:

[0461] Synthesis route:

[0462] Synthesis of intermediate 18-2:

[0463] Dissolve tert-butyldimethylhydroxyethoxysilane (18-1, 500 mg, 2.84 mmol) and di(p-nitrobenzene) carbonate (1.7 g, 5.67 mmol) in DMF (5 mL). Add DIEA (704 μL, 4.25 mmol) and allow to react at room temperature for 5 hours. After completion of the reaction as monitored by TLC, the product was purified by normal phase column chromatography (PE:EA = 10:1) to yield 18-2 as a pale yellow oil (640 mg, 66.1%). MS (ESI): m / z found [M+H] + =342.0.

[0464] Synthesis of intermediate 18-3:

[0465] Compound 11 (5 mg, 0.0118 mmol) was dissolved in 3 mL of dry DMF at room temperature. Compound 18-2 (4.1 mg, 0.0118 mmol), 1-hydroxybenzotriazole (1.8 mg, 0.01298 mmol), and N,N-diisopropylethylamine (6.2 μL, 0.0354 mmol) were added and reacted at room temperature for 4 hours. LC-MS monitoring showed that the starting material had reacted completely. The reaction solution was purified by pre-HPLC to yield compound 18-3 (4 mg, 54%). MS (ESI): m / z found [M+H] + =628.2.

[0466] Synthesis of compound 18:

[0467] Compound 18-3 (4 mg, 0.0064 mmol) was dissolved in 5 mL of dry DCM at room temperature, and trifluoroacetic acid (1 mL) was added. The mixture was allowed to react at room temperature for 1 hour. LC-MS monitoring showed that the reaction of the starting material was complete. The reaction solution was dried under reduced pressure and dissolved in a small amount of DMF and purified by pre-HPLC to obtain compound 18 (1.36 mg, 41%). MS (ESI): m / z found [M+H] + =514.35.

[0468] Example 19:

[0469] Synthesis route:

[0470] Synthesis of intermediate 19-2:

[0471] Boc-L-cyclopropylglycine (19-1, 500 mg, 2.32 mmol) was dissolved in EA (4 mL) and a 3M HCl / EA solution (7.4 mL) was added. The mixture was allowed to react overnight at room temperature. A large amount of white solid precipitated from the reaction mixture. The upper filter cake was filtered and dried to obtain a solid crude product 19-2 (320 mg). MS (ESI): m / z found [M+1] + =116.2.

[0472] Synthesis of intermediate 19-3:

[0473] Dissolve 19-2 in 2N dilute sulfuric acid (4.4 mL), cool in an ice bath, and add 4.4 mol / L sodium nitrite solution (5 mL) dropwise. Warm to room temperature and allow to react overnight. Saturate the solution with sodium chloride. Extract the filtrate with water / EA. Dry the organic phase over anhydrous sodium sulfate and spin dry to obtain a crude product (80 mg).

[0474] Synthesis of compound 19:

[0475] Compound 11 (15 mg, 0.036 mmol) was dissolved in DMF (2 mL) and cooled to 0°C in an ice bath. Intermediate 19-3 (9 mg, 0.072 mmol), DIEA (6 μL, 0.034 mmol), and HATU (5.3 mg, 0.014 mmol) were added and reacted in an ice bath for 0.5 hours. After LC-MS monitoring indicated the reaction was complete, the reaction solution was concentrated by vacuum distillation, purified by HPLC, and lyophilized to obtain a yellow solid product (0.82 mg, 8.9%). MS (ESI): m / z found [M+H] + =524.40.

[0476] Example 20:

[0477] Synthesis route:

[0478] Synthesis of intermediate 20-2:

[0479] Boc-D-cyclopropylglycine (20-1, 500 mg, 2.32 mmol) was dissolved in EA (4 mL) and a 3M HCl / EA solution (7.4 mL) was added. The mixture was allowed to react overnight at room temperature. A large amount of white solid precipitated from the reaction mixture. The upper layer was filtered and dried to obtain a solid crude product 20-2 (300 mg). MS (ESI): m / z found [M+1] + =116.2.

[0480] Synthesis of intermediate 20-3:

[0481] 20-2 was dissolved in 2N dilute sulfuric acid (4.4 mL), cooled in an ice bath, and 4.4 mol / L sodium nitrite solution (5 mL) was added dropwise. The temperature was raised to room temperature and the reaction was allowed to proceed overnight. Sodium chloride was added to saturate the solution, and the filtrate was extracted with water / EA, dried over anhydrous sodium sulfate, and spin-dried to give a crude product (90 mg).

[0482] Synthesis of compound 20:

[0483] 11 (20 mg, 0.047 mmol) was dissolved in DMF (3 mL) and cooled to 0°C in an ice bath. 20-3 (9 mg, 0.072 mmol), DIEA (6 μL, 0.034 mmol), and HATU (5.3 mg, 0.014 mmol) were added and reacted in an ice bath for 0.5 h. After LC-MS monitoring indicated the reaction was complete, the reaction solution was concentrated by vacuum distillation, purified by HPLC, and lyophilized to obtain a yellow solid product (1.44 mg, 15.6%). MS (ESI): m / z found [M+H] + =524.00. 1H NMR (600MHz, DMSO-d6) δ8.29(d,J=7.1Hz,1H),7.97(d,J=10.6Hz,1H),7.81(s,1H),6.71(d,J=2. 0Hz,1H),6.23(dd,J=8.6,6.4Hz,1H),5.92(dd,J=16.5,3.0Hz,1H),5.53(d,J=16.4Hz,3H),5.45( d,J=19.1Hz,1H),5.03(dd,J=16.6,13.4Hz,1H),4.93(tt,J=7.0,3.9Hz,1H),2.54(d,J=5.6Hz,3 H),2.05–1.95(m,2H),1.89(dd,J=9.1,6.0Hz,2H),1.72(d,J=12.6Hz,1H),0.86(t,J=7.3Hz,5H).

[0484] Example 21:

[0485] Synthesis route:

[0486] Synthesis of compound 21:

[0487] On ice, cis-3-hydroxycyclobutanecarboxylic acid (4.09 mg, 0.0353 mmol) was dissolved in 5 ml of dry DCM. N-hydroxysuccinimide (4.06 mg, 0.0353 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (6.77 mg, 0.0353 mmol) were added and allowed to react for one hour. Compound 11 (10.00 mg, 0.0235 mmol) and triethylamine (3.6 μL, 0.0259 mmol) were then added and the reaction was maintained at room temperature for two hours. The reaction was complete as monitored by LCMS. The reaction solution was purified by pre-HPLC to yield compound 21 (3.54 mg, 30% yield). MS (ESI): m / z found [M+H] + =524.3 1H NMR (400MHz, DMSO-d6) δ8.68(t,J=5.8Hz,1H),8.37(d,J=8.2Hz,1H),7.96(d,J=10.7Hz,1H),7.80(s,1H),6.71(s,1H),5.93(d,J=16.6Hz,1H),5.6 7(s,2H),5.53(d,J=16.7Hz,1H),5.09(d,J=7.0Hz,1H),4.89(d,J=5.8Hz ,2H),3.92(q,J=7.6Hz,1H),2.53(s,3H),2.03–1.97(m,7H),0.85(s,3H).

[0488] Example 22:

[0489] Synthesis route:

[0490] Synthesis of compound 22:

[0491] Compound 11 (15 mg, 0.035 mmol) was dissolved in 5 ml of dry DCM under ice. (S)-2-hydroxypropionic acid (6.4 mg, 0.071 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (16 mg, 0.042 mmol), and N,N-diisopropylethylamine (18 μL, 0.106 mmol) were added and the mixture was allowed to react at room temperature for 2 hours. LCMS monitored the reaction completion. The reaction solution was purified by pre-HPLC to give compound 22 (2.37 mg, 14% yield). MS (ESI): m / z found [M+H] + =498.35. 1 H NMR (600MHz, DMSO-d6) δ8.78(t,J=6.1Hz,1H),8.48(d,J=8.2Hz,1H),7.96(d,J=1 0.7Hz,1H),7.81(s,1H),6.71(s,1H),5.94(d,J=16.5Hz,1H),5.77(s,2H),5.63(d ,J=4.8Hz,1H),5.54(d,J=16.5Hz,1H),4.88(qd,J=15.1,6.1Hz,2H),4.06–3.98( m,1H),2.52(s,3H),2.00(dt,J=18.4,7.0Hz,3H),1.95–1.85(m,2H),0.86(s,3H).

[0492] Example 23:

[0493] Synthesis route:

[0494] Synthesis of compound 23:

[0495] Compound 11 (15 mg, 0.035 mmol) was dissolved in 5 ml of dry DCM under ice. Compound D-2-hydroxypropionic acid (6.4 mg, 0.071 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (16 mg, 0.042 mmol), and N,N-diisopropylethylamine (18 μL, 0.106 mmol) were added and the mixture was allowed to react at room temperature for 2 hours. LCMS monitored the reaction completion. The reaction solution was purified by pre-HPLC to give compound 23 (4.94 mg, 28% yield). MS (ESI): m / z found [M+H] + =498.35. 1 H NMR(600MHz,DMSO-d6)δ8.78(t,J=6.1Hz,1H),8.48(d,J=8.1Hz, 1H),7.95(s,1H),7.81(s,1H),6.71(s,2H),5.94(d,J=16.5Hz,1H),5.77(s,2H),5.63(d,J=4.8Hz,1H),5.54(d,J=16.5Hz,1H),4. 96–4.77(m,2H),4.09–3.96(m,1H),2.52(s,3H),2.00(dt,J=18.6,7.1Hz,3H),1.89(dq,J=14.3,7.1Hz,2H),0.86(d,J=4.5Hz,3H).

[0496] Example 24:

[0497] Synthesis route:

[0498] Synthesis of intermediate 24-1:

[0499] Compound 11 (100 mg, 0.235 mmol) was dissolved in 10 ml of dry N,N-dimethylformamide under an ice bath. Di-tert-butyl dicarbonate (56 mg, 0.259 mmol) and triethylamine (33 μL, 0.235 mmol) were added and the mixture was allowed to react at room temperature for 8 hours. LCMS confirmed the reaction was complete. The reaction solution was diluted with dichloromethane and washed three times with water. The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, and then dried to obtain compound 24-1 (140 mg, 99% yield). MS (ESI): m / z found [M+H] +=426.45.

[0500] Synthesis of intermediate 24-2:

[0501] Compound 24-1 (70 mg, 0.133 mmol) was dissolved in 10 ml of dry dichloromethane under an ice bath. Compound 6-2 (49 mg, 0.260 mmol), 4-dimethylaminopyridine (9.8 mg, 0.0798 mmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (122 mg, 638 mmol) were added and the mixture was allowed to react at room temperature for 1 hour. LCMS monitored the reaction completion. DMSO was added to the reaction solution until clear. Two volumes of water were then added to precipitate a solid. The solid was filtered and dried to obtain compound 24-2 (30 mg, 32% yield). MS (ESI): m / z found [M+H] + =696.40.

[0502] Synthesis of compound 24:

[0503] Compound 24-2 (30 mg, 0.043 mmol) was dissolved in 5 ml of dry dichloromethane at room temperature. Trifluoroacetic acid (2 ml) was added and the mixture was allowed to react at room temperature for 2 hours. LCMS confirmed the reaction was complete. The reaction solution was concentrated under reduced pressure and dissolved in a small amount of methanol. Pre-HPLC analysis yielded compound 24 (16.75 mg, 65% yield). MS (ESI): m / z found [M+H] + =596.36. 1 H NMR(600MHz,DMSO-d6)δ11.97(s,1H),8.45(d,J=8.0Hz,1H),8.36(s,2H),8.18(d,J=6.6Hz,1H), 8.00(d,J=10.5Hz,1H),7.73(s,1H),5.90(d,J=17.0Hz,1H),5.83(d,J=9.4Hz,2H),5.77(s,1H), 5.65(d,J=17.0Hz,1H),4.86(d,J=17.6Hz,1H),4.78(s,2H),3.17(s,3H),2.22(q,J=7.3Hz,2H),0.91(t,J=7.4Hz,3H).

[0504] Example 25:

[0505] Synthesis of compound 25:

[0506] Compound 24 (5 mg, 0.0084 mmol) was dissolved in 5 ml of dry dichloromethane under ice. Hydroxyacetic acid (1.3 mg, 0.0168 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (3.8 mg, 0.0101 mmol), and N,N-diisopropylethylamine (3.3 μL, 0.0252 mmol) were added and the mixture was allowed to react at room temperature for 2 hours. LCMS monitored the reaction completion. The reaction solution was concentrated under reduced pressure and dissolved in a small amount of N,N-dimethylformamide. Pre-HPLC purification afforded compound 25 (0.57 mg, 11% yield). MS (ESI): m / z found [M+H] + =654.35.

[0507] Example 26:

[0508] Synthesis route:

[0509] Synthesis of intermediate 26-2:

[0510] On an ice bath, glycolic acid (50 mg, 0.657 mmol) was dissolved in 5 ml of dry pyridine, tert-butyldiphenylsilyl trifluoromethanesulfonate (409 μL, 1.315 mmol) was added, and the mixture was allowed to react at room temperature for 8 hours. LCMS confirmed the reaction was complete. The reaction solution was concentrated under reduced pressure and dissolved in a small amount of methanol. The product was purified by reverse-phase column chromatography to obtain compound 26-2 (100 mg, 48% yield). MS (ESI): m / z found [MH] - =313.10.

[0511] Synthesis of intermediate 26-3:

[0512] Compound 11 (100 mg, 0.235 mmol) was dissolved in 5 ml of dry pyridine under ice, and triethylsilyl trifluoromethanesulfonate (504 μL, 2.35 mmol) was added. The mixture was then allowed to react at room temperature for 8 hours. LCMS confirmed the reaction was complete. The reaction solution was concentrated under reduced pressure and then dissolved in a small amount of N,N-dimethylformamide. The product was purified by reverse-phase column chromatography to obtain compound 26-3 (55 mg, 43% yield). MS (ESI): m / z found [M+H] + =540.40.

[0513] Synthesis of intermediate 26-4:

[0514] Compound 26-3 (55 mg, 0.102 mmol) was dissolved in 5 ml of dry dichloromethane under ice. Compound 26-2 (64 mg, 0.204 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (47 mg, 0.122 mmol), and N,N-diisopropylethylamine (54 μL, 0.306 mmol) were added and the mixture was allowed to react at room temperature for 1 hour. LCMS monitored the reaction completion. The reaction solution was concentrated under reduced pressure and purified by normal phase column chromatography to obtain compound 26-4 (60 mg, 70% yield). MS (ESI): m / z found [M+H] + =837.45.

[0515] Synthesis of intermediate 26-5:

[0516] Compound 26-4 (76 mg, 0.0909 mmol) was dissolved in 5 ml of toluene at room temperature. Lawesson's reagent (110 mg, 0.273 mmol) was added, and the atmosphere was replaced with nitrogen three times. The temperature was raised to 100°C and the reaction was allowed to react for 4 hours. TLC confirmed the reaction was complete. The reaction solution was concentrated under reduced pressure and purified by normal phase column chromatography to obtain compound 26-5 (43 mg, 55% yield).

[0517] Synthesis of compound 26:

[0518] Compound 26-5 (43 mg, 0.0505 mmol) was dissolved in 5 ml of dry dichloromethane at room temperature. Trifluoroacetic acid (2 ml) was added and the mixture was allowed to react at room temperature for 36 hours. LCMS confirmed the reaction was complete. The reaction solution was concentrated under reduced pressure and dissolved in a small amount of N,N-dimethylformamide. Pre-HPLC analysis yielded compound 26 (16.75 mg, 65% yield). MS (ESI): m / z found [M+H] + =500.35. 1 H NMR (600MHz, DMSO-d6) δ10.64(t,J=5.9Hz,1H),8.46(d,J=8.1Hz,1H),7.98(d,J=10.6Hz,1H),7.81(s,1H),6.71(s,1H),6.10(s,1H),5.93(d, J=16.5Hz,1H),5.70(s,2H),5.53(d,J=16.5Hz,1H),5.48(d,J=5.9Hz,2H),4.36(s,2H),2.52(s,3H),1.95–1.83(m,2H),0.86(t,J=7.3Hz,3H).

[0519] Example 27:

[0520] Synthesis route:

[0521] Synthesis of compound 27:

[0522] Compound 11 (15 mg, 0.22 mmol) was dissolved in DCM (2 mL). Glycolaldehyde dimer (7.62 mg, 0.40 mmol) and TEA (10.69 mg, 0.65 mmol) were added at 0°C under nitrogen protection, and the mixture was stirred for 0.5 h. NaBH(OAc)3 (14.96 mg, 0.44 mmol) was then added, and the temperature was slowly raised to room temperature for 12 h. LC-MS monitored the reaction completion, and the mixture was extracted with DCM / saturated sodium bicarbonate aqueous solution. The organic phase was dried over anhydrous Na2SO4, filtered, and spin-dried. Pre-HPLC analysis was performed and lyophilized to obtain compound 27 (1.2 mg, 7.25%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + =470.35. 1 H NMR (400MHz, DMSO-d6) δ8.39(d,J=8.3Hz,1H),8.19(s,1H),7.93(d,J=10.8Hz,1H),7.80(s,1H),6.71(s,1H),5.93(d,J=16.6Hz,1H),5.66(s,2H) ,5.53(d,J=16.6Hz,1H),4.41(s,2H),3.53(t,J=5.8Hz,2H),2.75–2.63( m,3H),2.53(s,3H),1.89(dt,J=10.1,6.6Hz,3H),0.86(t,J=7.3Hz,3H).

[0523] Example 28:

[0524] Synthesis of compound 28:

[0525] Compound 11 (15 mg, 0.035 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. 3-Hydroxycyclopentanecarboxylic acid (9.2 mg, 0.071 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (16 mg, 0.042 mmol), and N,N-diisopropylethylamine (18 μL, 0.106 mmol) were added and the mixture was allowed to react at room temperature for 2 hours. LCMS monitored the reaction to completion. The reaction solution was purified by pre-HPLC to give compound 28 (3.03 mg, 16% yield). MS (ESI): m / z found [M+H] + =538.40. 1H NMR (600MHz, DMSO-d6) δ8.75(t,J=5.9Hz,1H),8.37(d,J=8.2Hz,1H),7.96(d,J=10.7Hz,1H),7.80(s, 1H),6.72(s,1H),5.93(d,J=16.5Hz,1H),5.68(d,J=3.7Hz,2H),5.53(d,J=16.5Hz,1H),5.02(s,1H), 4.89(td,J=5.5,2.8Hz,2H),4.48(s,1H),2.87(p,J=8.4Hz,1H),2.53(s,3H),2.03–1.96(m,2H),1.94 –1.88(m,2H),1.70(ddd,J=8.6,6.9,4.4Hz,2H),1.46(dq,J=14.8,7.6,7.0Hz,2H),0.88–0.84(m,3H).

[0526] Example 29:

[0527] Synthesis route:

[0528] Synthesis of compound 29:

[0529] Compound 11 (10 mg, 0.023 mmol) was dissolved in DMF (1 mL) and cooled to 0°C in an ice bath. Compound 29-1 (4 mg, 0.047 mmol), TEA (8 μL, 0.059 mmol), and DMMTM (13 mg, 0.047 mmol) were added and reacted in an ice bath for 2 hours. After completion of the reaction as monitored by LC-MS, the product was purified by HPLC and lyophilized to afford the yellow solid product 29 (5.40 mg, 49.1%). MS (ESI): m / z found [M+H] + =486.30. 1 HNMR (600MHz, DMSO-d6) δ8.86(t,J=6.1Hz,1H),8.52(d,J=8.2Hz,1H),7.96(d,J=10.7Hz,1H),7.81(s,1H),6.72(s,1H),5.94(d,J=16.5 Hz,1H),5.79(s,2H),5.54(d,J=16.5Hz,1H),4.89(d,J=6.2Hz,2H),3.93(s,1H),2.53(s,3H),1.94–1.86(m,2H),0.86(t,J=7.3Hz,3H).

[0530] Example 30:

[0531] Synthesis of compound 30:

[0532] Compound 12 (5 mg, 0.011 mmol) was dissolved in dichloromethane (2 mL), cooled to 0°C in an ice bath, and cis-3-hydroxycyclobutanecarboxylic acid (3 mg, 0.023 mmol), DIEA (6 μL, 0.034 mmol), and HATU (5.3 mg, 0.014 mmol) were added. The mixture was allowed to react in an ice bath for 0.5 h. After completion of the reaction as monitored by LC-MS, the reaction solution was concentrated by vacuum distillation, purified by HPLC, and lyophilized to obtain the product 30 (2.4 mg, 40.7%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + =540.40. 1 H NMR (400MHz, DMSO-d6) δ8.65(t,J=6.0Hz,1H),8.04(d,J=12.1Hz,1H),7.91(d,J=9.2Hz,1H),7.77(s,1H),6.69(d,J=11.1Hz,2H),5.9 3(d,J=16.6Hz,1H),5.70(s,2H),5.52(d,J=16.6Hz,1H),5.32(t,J=4.8Hz,1H),4.96–4.86(m,2H),4.09(s,3H),3.91(q,J=7.7Hz,1H), 2.03–1.92(m,6H),0.87(d,J=7.3Hz,3H).

[0533] Example 31:

[0534] Synthesis of compound 31:

[0535] Compound 12 (5 mg, 0.011 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0°C in an ice bath. Glycolic acid (2 mg, 0.023 mmol), DIEA (6 μL, 0.034 mmol), and HATU (5.3 mg, 0.014 mmol) were added and reacted in an ice bath for 0.5 h. After the reaction was complete as monitored by LC-MS, the reaction solution was concentrated by vacuum distillation, purified by HPLC, and lyophilized to obtain the product 31 (0.77 mg, 14.3%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + =500.35. 1H NMR (400MHz, DMSO-d6) δ8.84(t,J=6.2Hz,1H),8.02(d,J=12.1Hz,1H),7.97(d,J=9.2Hz,1H),7.78(s,1H),6.74(s,1H),6.68(s,1H),5.93(d,J=16. 6Hz,1H),5.79(s,2H),5.50(d,J=16.6Hz,1H),5.31(t,J=4.9Hz,2H),4.9 5–4.88(m,2H),4.05(s,3H),2.00(d,J=7.5Hz,2H),0.84(d,J=4.7Hz,3H).

[0536] Example 32:

[0537] Synthesis of compound 32:

[0538] Compound 12 (5 mg, 0.011 mmol) was dissolved in dichloromethane (2 mL), cooled to 0°C in an ice bath, and L-lactic acid (2.1 mg, 0.023 mmol), DIEA (6 μL, 0.034 mmol), and HATU (5.3 mg, 0.014 mmol) were added. The mixture was reacted in an ice bath for 0.5 h. After LC-MS monitoring indicated the reaction was complete, the reaction solution was concentrated by vacuum distillation, purified by HPLC, and lyophilized to obtain the product 32 (0.61 mg, 10.9%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + =514.35. 1 H NMR (400MHz, DMSO-d6) δ8.80(t,J=6.1Hz,1H),8.04(d,J=12.1Hz,1H),7.98(d,J=9.2Hz,1H),7.78(s,1H),6.68(s,2H),5.93(d,J=16.6Hz,1H), 5.80(s,2H),5.53(d,J=16.5Hz,1H),5.32(t,J=4.8Hz,2H),4.96–4.85( m,2H),4.06(s,3H),2.00(d,J=7.8Hz,2H),0.86(dt,J=7.6,4.1Hz,6H).

[0539] Example 33:

[0540] Synthesis route:

[0541] Synthesis of intermediate 33-1;

[0542] 12 (15 mg, 0.034 mmol) was dissolved in DMF (2 mL), and 18-2 (12 mg, 0.034 mmol), HOBt (6 mg, 0.044 mmol), and DIEA (6 μL, 0.068 mmol) were added. The mixture was allowed to react at room temperature for 4 hours. After the reaction was complete as monitored by LC-MS, the mixture was purified by HPLC and lyophilized to obtain the yellow solid product 33-1 (12 mg, 54.5%). LC-MS (ESI): m / z found [M+H] + =644.40.

[0543] Synthesis of compound 33:

[0544] Compound 33-1 (12 mg, 0.019 mmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (1 mL) was added, and the mixture was allowed to react at room temperature for 1.5 hours. The reaction was complete after monitoring by LC-MS. The mixture was purified by HPLC and lyophilized to obtain a yellow solid product 33 (4.76 mg, 47.6%). LC-MS (ESI): m / z found [M+H] + =530.35. 1 H NMR (600MHz, DMSO-d6) δ8.22(t,J=6.1Hz,1H),8.04(d,J=12.0Hz,1H),7.87(d,J= 9.1Hz,1H),7.77(s,1H),6.70(s,1H),5.93(d,J=16.4Hz,1H),5.74(s,2H),5.52( d,J=16.4Hz,1H),4.83(d,J=6.1Hz,2H),4.72(s,1H),4.11(s,3H),3.99(t,J=5.1 Hz,2H),3.54(t,J=5.1Hz,2H),1.94–1.84(m,J=7.2Hz,2H),0.87(t,J=7.3Hz,3H).

[0545] Example 34:

[0546] Synthesis of compound 34:

[0547] Compound 12 (5 mg, 0.011 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0°C in an ice bath. D-lactic acid (2.1 mg, 0.023 mmol), DIEA (6 μL, 0.034 mmol), and HATU (5.3 mg, 0.014 mmol) were added and reacted in an ice bath for 0.5 h. After completion of the reaction as monitored by LC-MS, the reaction solution was concentrated by vacuum distillation, purified by HPLC, and lyophilized to obtain the product 34 as a yellow solid (1.00 mg, 17.9%). LC-MS (ESI): m / z found [M+H] + =514.35. 1 H NMR(600MHz,DMSO-d6)δ8.80(t,J=6.2Hz,1H),8.04(d,J=12.0Hz,1H),7.99(d,J=9.2Hz ,1H),7.78(s,1H),6.70(s,1H),5.94(d,J=16.5Hz,1H),5.86–5.74(m,2H),5.61(d,J=4. 9Hz,1H),5.53(d,J=16.4Hz,1H),4.90(ddd,J=50.7,15.0,6.2Hz,2H),4.07(s,3H),4.04 (td,J=6.7,4.4Hz,1H),1.95–1.83(m,J=7.3Hz,2H),1.23(s,3H),0.87(t,J=7.3Hz,3H).

[0548] Example 35:

[0549] Synthesis route:

[0550] Synthesis of intermediate 35-1:

[0551] 12 (100 mg, 0.180 mmol) was dissolved in dichloromethane (3 mL), cooled to 0°C in an ice bath, and 26-2 (113 mg, 0.360 mmol), DIEA (89 μL, 0.540 mmol), and HATU (82 mg, 0.216 mmol) were added. The mixture was allowed to react at room temperature for 2 hours. After the reaction was complete as monitored by LC-MS, the reaction solution was concentrated by distillation under reduced pressure and purified by TLC (DCM:EA=1:1) to obtain the product 35-1 (90 mg, 58.8%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + =852.60.

[0552] Synthesis of intermediate 35-2:

[0553] Under nitrogen, compound 35-1 (90 mg, 0.106 mmol) was dissolved in toluene (3 mL), and Lawesson's reagent (171 mg, 0.424 mmol) was added. The mixture was reacted at 100°C for 4 hours. After completion of the reaction as monitored by TLC, the mixture was evaporated to dryness under reduced pressure and purified by TLC (PE:EA = 5:1) to afford the product 35-2 (45 mg, 48.9%) as a yellow solid.

[0554] Synthesis of compound 35:

[0555] Compound 35-2 (45 mg, 0.052 mmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (5 mL) was added, and the mixture was allowed to react at room temperature for 3 days. After the reaction was complete as monitored by LC-MS, the mixture was concentrated by vacuum distillation, prepared by HPLC, and lyophilized to obtain a yellow solid product 35 (3.34 mg, 12.5%). LC-MS (ESI): m / z found [M+H] + =516.30. 1 HNMR(600MHz,DMSO-d6)δ10.59(t,J=5.9Hz,1H),8.06(d,J=11.9Hz,1H),7.85(d,J=9.0Hz,1H),7.79(s,1H) ,6.70(s,1H),6.05(t,J=5.8Hz,1H),5.93(d,J=16.4Hz,1H),5.76(d,J=2.9Hz,2H),5.53(d,J=16.5Hz,1H), 5.50(dd,J=10.3,5.9Hz,1H),4.35(d,J=5.7Hz,2H),4.06(s,3H),2.55(s,1H),1.90(hept,J=7.1Hz,2H),0.87(t,J=7.4Hz,3H).

[0556] Example 36:

[0557] Synthesis of compound 36:

[0558] Compound 13 (5 mg, 0.011 mmol) was dissolved in 5 ml of dry dichloromethane under ice. Hydroxyacetic acid (1.7 mg, 0.022 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (5 mg, 0.013 mmol), and N,N-diisopropylethylamine (5.7 μL, 0.033 mmol) were added and the mixture was allowed to react at room temperature for 2 hours. LCMS monitored the reaction to completion. The reaction solution was purified by pre-HPLC to give compound 36 (1.55 mg, 28% yield). MS (ESI): m / z found [M+H] +=510.35. 1 HNMR(600MHz,DMSO-d6)δ8.80(t,J=6.1Hz,1H),7.97(s,1H),7.76(s,1H),7.62(s,1H),6.69(s,1H),5.93(d,J=16.4Hz,1H),5. 77(d,J=1.9Hz,3H),5.52(d,J=16.4Hz,1H),4.79(dd,J=6.1,2.8Hz,2H),4.46(s,6H),1.92–1.88(m,2H),0.86(t,J=7.3Hz,3H).

[0559] Example 37:

[0560] Synthesis route:

[0561] Synthesis of intermediate 37-1:

[0562] Compound 13 (15 mg, 0.033 mmol) was dissolved in 3 ml of dry N,N-dimethylformamide at room temperature. Compound 18-2 (11 mg, 0.033 mmol), 1-hydroxybenzotriazole (5 mg, 0.036 mmol), and N,N-diisopropylethylamine (17 μL, 0.099 mmol) were added and the mixture was allowed to react at room temperature for 3 hours. LCMS monitored the reaction for completion. The reaction solution was purified by pre-HPLC to afford compound 37-1 (23 mg, 99% yield). MS (ESI): m / z found [M+H] + =654.45.

[0563] Synthesis of compound 37:

[0564] Compound 37-1 (23 mg, 0.035 mmol) was dissolved in 5 ml of dry dichloromethane at room temperature. Trifluoroacetic acid (2 ml) was added and the mixture was allowed to react at room temperature for 2 hours. LCMS confirmed the reaction was complete. The reaction solution was concentrated under reduced pressure and dissolved in a small amount of N,N-dimethylformamide. Pre-HPLC purification afforded compound 37 (4.21 mg, 22% yield). MS (ESI): m / z found [M+H] + =540.35. 1H NMR (400MHz, DMSO-d6) δ8.84(t,J=6.2Hz,1H),8.02(d,J=12.1Hz,1H),7.97(d,J=9.2Hz,1H),7.78(s,1H),6.74(s,1H),6.68(s,1H),5.93(d,J=16. 6Hz,1H),5.79(s,2H),5.50(d,J=16.6Hz,1H),5.31(t,J=4.9Hz,2H),4.9 5–4.88(m,2H),4.05(s,3H),2.00(d,J=7.5Hz,2H),0.84(d,J=4.7Hz,3H).

[0565] Example 38:

[0566] Synthesis of compound 38:

[0567] Compound 13 (5 mg, 0.011 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. cis-3-hydroxycyclobutanecarboxylic acid (2.6 mg, 0.022 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (4.9 mg, 0.013 mmol), and N,N-diisopropylethylamine (5.7 μL, 0.033 mmol) were added and the mixture was allowed to react at room temperature for 2 hours. LCMS monitored the reaction to completion. The reaction solution was purified by pre-HPLC to give compound 38 (0.34 mg, 5% yield). MS (ESI): m / z found [M+H] + =550.45. 1 H NMR(400MHz,DMSO-d6)δ8.62(t,J=5.9Hz,1H),7.80(s,1H),7.75(s,1H),7.61(s,1H) ,6.68(s,1H),5.93(d,J=16.5Hz,1H),5.64(s,2H),5.51(d,J=16.6Hz,1H),5.09(s,1 H),4.76(d,J=5.8Hz,2H),4.46(s,4H),3.91(t,J=7.9Hz,1H),2.45–2.36(m,1H),2.2 7(qt,J=7.4,3.8Hz,2H),1.90(dtd,J=18.1,10.5,4.4Hz,4H),0.86(t,J=7.3Hz,3H).

[0568] Example 39:

[0569] Synthesis route:

[0570] Synthesis of intermediate 39-1:

[0571] Compound 13-8 (60 mg, 0.106 mmol) was dissolved in 5 ml of dry dichloromethane under ice. Compound 26-2 (67 mg, 0.212 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (48 mg, 0.127 mmol), and N,N-diisopropylethylamine (55 μL, 0.318 mmol) were added and the mixture was allowed to react at room temperature for 1 hour. LCMS monitored the reaction completion. The reaction solution was concentrated under reduced pressure and purified by normal phase column chromatography to obtain compound 39-1 (53 mg, 58%). MS (ESI): m / z found [M+H] + =862.55.

[0572] Synthesis of intermediate 39-2:

[0573] Compound 39-1 (53 mg, 0.061 mmol) was dissolved in 5 ml of toluene at room temperature. Lawesson's reagent (75 mg, 0.184 mmol) was added, and the atmosphere was replaced with nitrogen three times. The temperature was raised to 100°C and the reaction was allowed to react for 4 hours. TLC confirmed the reaction was complete. The reaction solution was concentrated under reduced pressure and purified by normal phase column chromatography to obtain compound 39-2 (37 mg, 69% yield).

[0574] Synthesis of compound 39:

[0575] Compound 39-2 (43 mg, 0.0505 mmol) was dissolved in 5 ml of dry dichloromethane at room temperature. Trifluoroacetic acid (2 ml) was added and the mixture was allowed to react at room temperature for 48 hours. LCMS confirmed the completion of the reaction. The reaction solution was concentrated under reduced pressure and dissolved in a small amount of N,N-dimethylformamide. Pre-HPLC analysis yielded compound 39 (4.08 mg, 18% yield). MS (ESI): m / z found [M+H] + =526.35. 1 HNMR(600MHz,DMSO-d6)δ10.55(t,J=5.8Hz,1H),7.89(s,1H),7.76(s,1H),7.63(s,1H),6.67(s,1H),6.07(t,J=5.7Hz,1H),5.92(d,J=16.4Hz,1H) ,5.71(s,2H),5.52(d,J=16.4Hz,1H),5.35(dd,J=5.9,3.4Hz,2H),4.46(s ,4H),4.32(d,J=5.4Hz,2H),1.90(hept,J=7.1Hz,2H),0.88–0.85(m,3H).

[0576] Example 40:

[0577] Compound 13 (5 mg, 0.011 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. (S)-2-hydroxypropionic acid (1.6 μL, 0.022 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (4.9 mg, 0.013 mmol), and N,N-diisopropylethylamine (5.7 μL, 0.033 mmol) were added and the mixture was allowed to react at room temperature for 2 hours. LCMS monitored the reaction to be complete. The reaction solution was purified by pre-HPLC to afford compound 40 (0.34 mg, 5% yield). MS (ESI): m / z found [M+H] + =524.40. 1 H NMR(600MHz,DMSO-d6)δ8.73(t,J=6.1Hz,1H),7.94(s,1H),7.75(s,1H),7.62(d,J =10.8Hz,1H),6.67(s,1H),5.92(s,1H),5.75(d,J=3.7Hz,2H),5.60(d,J=4.9Hz,1 H),5.52(d,J=16.4Hz,2H),4.82(d,J=6.4Hz,1H),4.77(dd,J=8.6,6.1Hz,2H),4.4 6(s,4H),4.03–3.95(m,1H),1.91–1.88(m,2H),1.22(d,J=6.8Hz,3H),0.86(s,3H).

[0578] Example 41:

[0579] Compound 13 (5 mg, 0.011 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. D-2-hydroxypropionic acid (1.6 μL, 0.022 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (4.9 mg, 0.013 mmol), and N,N-diisopropylethylamine (5.7 μL, 0.033 mmol) were added and the mixture was allowed to react at room temperature for 2 hours. LCMS monitored the reaction to completion, and the reaction solution was purified by pre-HPLC to afford compound 41 (1.16 mg, 20% yield). MS (ESI): m / z found [M+H] + =524.40. 1H NMR(600MHz,DMSO-d6)δ8.72(t,J=6.1Hz,1H),7.94(s,1H),7.75(s,1H),7.61(d, J=1.2Hz,1H),6.67(s,1H),5.93(d,J=16.4Hz,1H),5.75(s,2H),5.60(d,J=4.9Hz, 1H),5.52(d,J=16.4Hz,1H),4.77(d,J=6.0Hz,2H),4.46(s,4H),4.00(q,J=6.7,6 .1Hz,1H),1.90(hept,J=7.3Hz,2H),1.21(d,J=6.7Hz,3H),0.86(t,J=7.3Hz,3H).

[0580] Example 42:

[0581] Compound 14 (5 mg, 0.0113 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. cis-3-hydroxycyclobutanecarboxylic acid (2.6 mg, 0.0226 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (5.2 mg, 0.0136 mmol), and N,N-diisopropylethylamine (5.9 μL, 0.0339 mmol) were added and the mixture was allowed to react at room temperature for 2 hours. LCMS monitored the reaction completion. The reaction solution was purified by pre-HPLC to give compound 42 (3.03 mg, 50% yield). MS (ESI): m / z found [M+H] + =540.03. 1 H NMR(600MHz,DMSO-d6)δ8.68(t,J=5.9Hz,1H),8.41(s,1H),8.32(s,1H),7 .80(s,1H),6.71(s,1H),5.93(d,J=16.5Hz,1H),5.68(s,2H),5.53(d,J=1 6.5Hz,1H),5.09(d,J=7.0Hz,1H),4.89(d,J=5.9Hz,2H),3.92(q,J=7.1Hz ,1H),2.61(s,3H),2.44(tt,J=9.8,7.4Hz,1H),2.29(ddt,J=11.7,6.9,3.6 Hz,2H),1.98–1.92(m,2H),1.89(dt,J=14.3,7.5Hz,2H),0.86(d,J=7.4Hz,3H).

[0582] Example 43:

[0583] Compound 14 (5 mg, 0.0113 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. (S)-2-hydroxypropionic acid (1.7 μL, 0.0226 mmol), 4-(4,6-dimethoxytriazin-2-yl)-4-methylmorpholine hydrochloride (6.3 mg, 0.0226 mmol), and triethylamine (3.9 μL, 0.0283 mmol) were added and the mixture was allowed to react at room temperature for 1 hour. LCMS monitored the reaction completion. The reaction solution was purified by pre-HPLC to afford compound 43 (1.76 mg, 30% yield). MS (ESI): m / z found [M+H] + =514.35. 1 HNMR(400MHz,DMSO-d6)δ8.78(t,J=6.0Hz,1H),8.51(s,1H),8.31(s,1H),7.8 0(s,1H),6.71(s,1H),5.93(d,J=16.7Hz,1H),5.77(s,2H),5.63(d,J=4.8Hz, 1H),5.53(d,J=16.7Hz,1H),4.97–4.81(m,2H),4.06–3.95(m,1H),2.60(s,3H ), 1.90 (dt, J = 11.6, 6.9 Hz, 2H), 1.22 ( d, J = 6.8 Hz, 4H), 0.86 ( t, J = 7.3 Hz, 3H).

[0584] Example 44:

[0585] Compound 15 (5 mg, 0.011 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. cis-3-hydroxycyclobutanecarboxylic acid (2.6 mg, 0.022 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (4.9 mg, 0.013 mmol), and N,N-diisopropylethylamine (5.7 μL, 0.033 mmol) were added and the mixture was allowed to react at room temperature for 2 hours. LCMS monitored the reaction to completion. The reaction solution was purified by pre-HPLC to give compound 44 (0.34 mg, 5% yield). MS (ESI): m / z found [M+H] + =550.45. 1HNMR(600MHz,DMSO-d6)δ8.69(t,J=6.0Hz,1H),8.50(dd,J=12.3,8.6Hz,1H),8.30(dd, J=11.3,8.0Hz,1H),7.81(s,1H),6.72(s,1H),5.94(d,J=16.5Hz,1H),5.72(s,2H),5.5 4(d,J=16.5Hz,1H),5.09(d,J=7.0Hz,1H),4.86(d,J=6.0Hz,2H),3.97–3.86(m,1H),2. 43(tt,J=9.9,7.5Hz,1H),2.32–2.23(m,2H),1.96–1.85(m,4H),0.86(t,J=7.3Hz,3H).

[0586] Example 45:

[0587] Compound 15 (5 mg, 0.0116 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. (S)-2-hydroxypropionic acid (1.7 μL, 0.0232 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (5.3 mg, 0.0139 mmol), and N,N-diisopropylethylamine (6.1 μL, 0.0348 mmol) were added and the mixture was allowed to react at room temperature for 1 hour. LCMS monitored the reaction completion. The reaction solution was purified by pre-HPLC to give compound 45 (1.75 mg, 30% yield). MS (ESI): m / z found [M+H] + =502.30. 1 HNMR(600MHz,DMSO-d6)δ8.85(t,J=6.1Hz,1H),8.63(dd,J=12.4,8.6Hz,1H),8.3 0(dd,J=11.3,8.0Hz,1H),7.82(s,1H),6.72(s,1H),5.94(d,J=16.6Hz,1H),5.81( s,2H),5.64(d,J=4.8Hz,1H),5.54(d,J=16.5Hz,1H),4.89–4.80(m,2H),4.04–3. 97(m,1H),1.90(hept,J=7.1Hz,2H),1.20(d,J=6.8Hz,3H),0.87(t,J=7.3Hz,3H).

[0588] Example 46:

[0589] Synthesis route:

[0590] Synthesis of intermediate 46-3:

[0591] At room temperature, 5-bromo-4-fluoro-2-nitrobenzaldehyde (46-1, 1.0 g, 4.032 mmol) was dissolved in 20 mL of toluene. T-butyl carbamate (567 mg, 4.839 mmol), Pd2(dba)3 (185 mg, 0.202 mmol), CsCO3 (4.2 g, 8.064 mmol), and Xphos (385 mg, 0.801 mmol) were added. The atmosphere was purged with nitrogen three times, then the temperature was raised to 90°C and the reaction was allowed to proceed overnight. LC-MS monitoring indicated complete reaction of the starting material. The reaction solution was diluted with ethyl acetate, washed three times with water, and the combined organic phases were dried over anhydrous Na2SO4, concentrated, and isolated by column chromatography (petroleum ether:ethyl acetate = 93:7) to afford compound 46-3 (878 mg, 77% yield). LC-MS (ESI): m / z found [M+H] + =285.2.

[0592] Synthesis of intermediate 46-4:

[0593] Compound 46-4 (878 mg, 3.089 mmol) was dissolved in 5 mL of water and 20 mL of ethanol at room temperature. Ammonium chloride (268 mg, 5.004 mmol) and iron powder (691 mg, 12.356 mmol) were added. The atmosphere was purged with nitrogen three times, then the temperature was raised to 80°C and the reaction was allowed to proceed overnight. LC-MS monitoring indicated that the starting material had reacted completely. The reaction mixture was filtered to remove the iron powder, and the filtrate was concentrated and diluted with ethyl acetate. The organic phases were washed three times with water, combined, dried over anhydrous Na2SO4, and separated by column chromatography (dichloromethane:methanol = 99:1) to afford compound 46-4 (571 mg, 73% yield). MS (ESI): m / z found [M+H] + =255.4.

[0594] Synthesis of intermediate 46-5:

[0595] At room temperature, compound 46-4 (571 mg, 2.242 mmol) was dissolved in 10 mL of toluene, and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyran-O-[3,4-F]-indolizine-3,6,10(4H)-one (492 mg, 1.868 mmol) and TsOH (33 mg, 0.187 mmol) were added. The temperature was raised to 110°C and the reaction was allowed to proceed for 2 hours. LC-MS monitoring showed that the reaction of the starting material was complete. The reaction solution was cooled and concentrated, and then recrystallized by adding diethyl ether to obtain compound 46-5 (749 mg, yield 69%). LC-MS (ESI): m / z found [M+H] + =482.

[0596] Synthesis of intermediate 46-6:

[0597] Compound 46-5 (400 mg, 0.831 mmol) was dissolved in 6 mL of dry pyridine at room temperature, and TESOTf (1.8 mL, 8.31 mmol) was added and allowed to react overnight. TLC monitored the reaction for completion. The reaction solution was dried, diluted with DCM, and washed three times with water and once with a saturated aqueous NaCl solution. After drying over anhydrous Na2SO4, the solution was concentrated and purified by column chromatography (petroleum ether:ethyl acetate = 65:35) to afford compound 46-6 (340 mg, 68% yield). MS (ESI): m / z found [M+H] + =596.2.

[0598] Synthesis of intermediate 46-7:

[0599] At room temperature, compound 46-6 (202 mg, 0.339 mmol) was dissolved in 10 mL of toluene, and Lawesson's reagent (343 mg, 0.848 mmol) was added. The temperature was raised to 100°C and the reaction was allowed to proceed for 4 hours. The reaction was monitored to be complete by TLC. The reaction solution was concentrated and dissolved in a small amount of dichloromethane and purified by column chromatography (petroleum ether:ethyl acetate = 65:35) to obtain compound 46-7 (102 mg, yield 95%).

[0600] Synthesis of compound 46:

[0601] Compound 46-7 (30 mg, 0.049 mmol) was dissolved in 5 ml of dry DCM at room temperature, and trifluoroacetic acid (1 ml) was added. The reaction was allowed to react overnight at room temperature. LC-MS monitored the reaction to be complete. The reaction solution was purified by reverse phase column chromatography to obtain compound 46 (20 mg, 99% yield). LC-MS (ESI): m / z found [M+H] + =398.0.

[0602] Example 47:

[0603] Synthesis route:

[0604] Synthesis of intermediate 47-2:

[0605] Compound 47-1 (2.0 g, 6.51 mmol) was dissolved in MeOH (40 mL), and NaBH4 (0.172 g, 4.55 mmol) was added in three portions. The mixture was allowed to react at room temperature for 30 min. The solvent was removed by distillation under reduced pressure, and a small amount of H2O was added to dissolve the mixture. 2M HCl aqueous solution was added dropwise to adjust the pH to 4. A large amount of solid precipitated. The solid was filtered, and the filter cake was washed with a small amount of cold water and lyophilized to obtain the off-white solid product 47-2 (1.135 g, 56.4% yield). LC-MS: [M+H] + =310.2.

[0606] Synthesis of intermediate 47-3:

[0607] To compound 47-2 (1.135 g, 3.67 mmol) was added 60 mL of glacial acetic acid. NaIO4 (1.138 g, 5.32 mmol) was dissolved in 12 mL of H2O. The aqueous NaIO4 solution was added dropwise to the glacial acetic acid solution of compound 47-2 at room temperature and stirred at room temperature until the reaction was complete. The reaction was quenched with 1 mL of EG and extracted with DCM / H2O. The mixture was dried over anhydrous Na2SO4 and filtered. The solvent was removed by distillation under reduced pressure and the product was purified by medium pressure liquid chromatography (DCM:MeOH = 95:5). Concentration afforded the product 47-3 (1.0 g, 88.9% yield) as a white solid. LC-MS: [M+H] + =308.0.

[0608] Synthesis of intermediate 47-4:

[0609] A suspension of Zn (1.625 g, 24.7 mmol) in anhydrous THF (30 mL) was stirred under N₂ protection. TMSCl (0.706 g, 6.5 mmol) was added dropwise at room temperature for activation for 15 min, then the mixture was heated to reflux. The heating bath was removed and tert-butyl bromoacetate (3.6 mL, 24.7 mmol) was added dropwise while maintaining reflux. The mixture was heated to reflux until the zinc powder was completely dissolved. A solution of 47-3 in anhydrous THF was then added dropwise, and the mixture was heated to reflux for 2 h. After cooling to room temperature, the mixture was extracted with DCM / saturated aqueous NH₄Cl, dried over anhydrous Na₂SO₄, and filtered. The solvent was removed by distillation under reduced pressure, and the mixture was purified by medium-pressure liquid chromatography (DCM:MeOH = 96:6) and concentrated to afford 47-4 as a pale yellow oil (1.268 g, 98.6% yield). LC-MS: [M+H] + =396.0.

[0610] Synthesis of intermediate 47-5:

[0611] Trifluoroacetic acid (7.74 mL) was added dropwise to 47-4 (1.268 g, 3.206 mmol) at room temperature and allowed to react overnight. The solvent was removed by distillation under reduced pressure and the product was purified by medium-pressure liquid chromatography (DCM:MeOH = 98:2). Concentration afforded 47-5 (0.502 g, 56.5% yield) as a light pink solid. LC-MS: [M+H] + =278.0.

[0612] Synthesis of intermediate 47-6:

[0613] At room temperature, 47-5 (0.251 g, 0.905 mmol) was dissolved in 3 mL of toluene and 3 mL of glacial acetic acid. Compound 11-3 (0.182 g, 0.905 mmol) and p-TsOH·H2O (9 mg) were then added sequentially. The temperature was raised to 110°C and the reaction was allowed to proceed overnight. After removing the solvent by distillation under reduced pressure, 8 mL of acetone was added, and the mixture was stirred at room temperature for 30 min. The mixture was filtered to obtain a light yellow solid 47-6 (0.247 g, 63.4% yield). LC-MS: [M+H] + =443.0.

[0614] Synthesis of compound 47:

[0615] Compound 47-6 (50 mg, 0.113 mmol) was dissolved in 5 mL of EtOH. Hexamethylenetetramine (47.6 mg, 0.339 mmol) was added at room temperature and the mixture was heated to 80°C and refluxed for 48 h. The solvent was removed by distillation under reduced pressure and the mixture was purified by reverse-phase column chromatography to obtain the yellow solid product 47 (10 mg, 20.9% yield). LC-MS: [M+H] + =424.0.

[0616] Example 48:

[0617] Synthesis route:

[0618] Synthesis of intermediate 48-2:

[0619] 25 mL of DCE was added to an eggplant-shaped flask, cooled to 0°C in an ice bath, and BCl₃ (6 mL, 6.39 mmol) was added. After stirring on ice for 10 min, 11-1 (1 g, 7.99 mmol), 48-1 (1.502 g, 9.388 mmol), and AlCl₃ (1.39 g, 10.4 mmol) were added sequentially. The mixture was allowed to react at room temperature for 10 min, then heated to 80°C and allowed to react overnight. The reaction solution was cooled to room temperature, and H₂O (20 mL) was slowly added in an ice bath. After stirring for 10 min, 2M aqueous HCl (40 mL) was added and stirred at 0°C for 30 min. Extraction was performed with DCM / H₂O (30 mL × 3), dried over anhydrous Na₂SO₄, and filtered. The solvent was removed by distillation under reduced pressure, and the mixture was purified by medium-pressure liquid chromatography (petroleum ether:ethyl acetate = 2:1) to afford 48-2 (1.85 g, 80.4% yield), a red oil. LC-MS: [M+H] + =288.0.

[0620] Synthesis of intermediate 48-3:

[0621] At room temperature, 48-2 (100 mg, 0.361 mmol) was dissolved in 3 mL of toluene and 3 mL of glacial acetic acid. Compound 47-5 (83 mg, 0.288 mmol) and p-TsOH·H2O (5 mg) were then added sequentially. The temperature was raised to 110°C and the reaction mixture was allowed to react for 7 h. The solvent was removed by distillation under reduced pressure, and the mixture was purified by medium-pressure liquid chromatography to obtain compound 48-3 (30 mg, 15.7% yield). LC-MS: [M+H] + =529.0.

[0622] Synthesis of compound 48:

[0623] Compound 48-3 (30 mg, 0.057 mmol) was dissolved in HMPA (1.5 mL) and H2O (0.3 mL), and the temperature was raised to 110°C for 4 h. HPLC preparation was performed to obtain compound 27 (2.26 mg, yield 8.5%). LC-MS: [M+H] + =467.35. 1H NMR (600MHz, DMSO-d6) δ8.24(d,J=8.2Hz,1H),7.85(d,J=10.8Hz,1H),7.37(s,1H),6.04(s,1H ),5.53(d,J=15.1Hz,1H),5.40(d,J=15.0Hz,1H),5.28(d,J=4.1Hz,2H),4.45(t,J=5.1Hz,1H) ,3.48(q,J=6.8,5.9Hz,3H),3.20(dq,J=9.2,6.0,5.1Hz,2H),3.05(d,J=13.9Hz,1H),2.52(s, 3H), 1.85 (q, J = 7.4Hz, 2H), 1.74 (p, J = 7.9Hz, 2H), 1.62 (p, J = 6.4Hz, 2H), 0.86 (t, J = 7.4Hz, 4H).

[0624] Example 49:

[0625] Synthesis route:

[0626] Synthesis of intermediate 49-1:

[0627] At room temperature, 47-5 (100 mg, 0.361 mmol) was dissolved in 3 mL of toluene and 3 mL of glacial acetic acid. Compound 15-2 (66.6 mg, 0.325 mmol) and p-TsOH·H2O (5 mg) were then added sequentially. The temperature was raised to 110°C and the reaction mixture was allowed to react for 5 h. The solvent was removed by distillation under reduced pressure, and the mixture was purified by medium-pressure liquid chromatography (dichloromethane:methanol = 94:6) to obtain 49-1 (50 mg, 31.0% yield) as a yellow solid. LC-MS: [M+H] + =447.0.

[0628] Synthesis of compound 49:

[0629] Compound 49-1 (50 mg, 0.112 mmol) was dissolved in 5 mL of EtOH. Hexamethylenetetramine (46 mg, 0.336 mmol) was added at room temperature and the mixture was heated to 78°C and refluxed for 48 h. The solvent was removed by distillation under reduced pressure and purified by HPLC to obtain product 49 as a pale yellow solid (1 mg, 2.1% yield). LC-MS: [M+H] + =428.0.

[0630] Example 50:

[0631] Synthesis route:

[0632] Synthesis of compound 50:

[0633] Compound 1 (5 mg, 0.0050 mmol) was dissolved in ultra-dry DMF under an ice-water bath. 50-1 (3.5 mg, 0.006 mmol), HATU (2.5 mg, 0.0065 mmol), HOBt (0.9 mg, 0.0065 mmol), and N,N-diisopropylethylamine (1.5 mg, 0.011 mmol) were added sequentially. The mixture was allowed to react at room temperature for one hour. LCMS monitored the reaction to be complete. The reaction solution was concentrated and purified by Pre-HPLC to give compound 50 (1.0 mg, 12%). MS (ESI): m / z found [M+H] + =1475.4.

[0634] Example 51:

[0635] Synthesis route:

[0636] Synthesis of compound 51:

[0637] Under ice-water bath, 2 (4.09 mg, 0.0069 mmol) was dissolved in ultra-dry DMF (5 mL). 50-1 (4.28 mg, 0.0069 mmol), HATU (4.56 mg, 0.0079 mmol), HOBt (1.07 mg, 0.0079 mmol), and N,N-diisopropylethylamine (2.00 mg, 0.0150 mmol) were added sequentially. The mixture was allowed to react at room temperature for one hour. LC-MS monitored the reaction to be complete. The reaction solution was concentrated and purified by Pre-HPLC to give compound L1-S02 (5.90 mg, 73%). MS (ESI): m / z found [M / 2+H] + =648.3.

[0638] Example 52:

[0639] Synthesis route:

[0640] Synthesis of compound 52:

[0641] Under ice-water bath, 3 (5.0 mg, 0.0068 mmol) was dissolved in ultra-dry DMF (5 mL). 50-1 (4.2 mg, 0.0068 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (4.5 mg, 0.0078 mmol), 1-hydroxybenzotriazole monohydrate (1.1 mg, 0.0078 mmol), and N,N-diisopropylethylamine (1.7 mg, 0.0129 mmol) were added sequentially. The mixture was allowed to react for 10 minutes and then at room temperature for 1 hour. LC-MS monitored the reaction to determine completion. The reaction solution was concentrated and purified by Pre-HPLC to give compound L1-3 (2.43 mg, 27%). MS (ESI): m / z found [M+H] + =1331.5.

[0642] Example 53:

[0643] Synthesis route:

[0644] Synthesis of compound 53:

[0645] Under ice-water bath, 6 (5.0 mg, 0.0083 mmol) was dissolved in ultra-dry DMF. 50-1 (5.1 mg, 0.0083 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (5.5 mg, 0.0095 mmol), 1-hydroxybenzotriazole monohydrate (1.3 mg, 0.007895 mmol), and N,N-diisopropylethylamine (2.1 mg, 0.0158 mmol) were added sequentially. The mixture was allowed to react for 10 minutes and then at room temperature for 1 hour. LC-MS monitored the reaction to be complete. The reaction solution was concentrated and purified by Pre-HPLC to yield the compound (4.08 mg, 41%). MS (ESI): m / z found [M+H] + =1205.4.

[0646] Example 54:

[0647] Synthesis route:

[0648] Synthesis of compound 54:

[0649] Compound 7 (5 mg, 0.0077 mmol) was dissolved in 2 mL of dry DMF at room temperature under an ice bath. 50-1 (4.7 mg, 0.0077 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (5.1 mg, 0.0088 mmol), 1-hydroxybenzotriazole monohydrate (1.2 mg, 0.0088 mmol), and N,N-diisopropylethylamine (1.9 mg, 0.0146 mmol) were added sequentially. The mixture was allowed to react for 10 minutes, then the temperature was raised to room temperature and the reaction was continued for 2 hours. LC-MS monitored the reaction completion. The reaction solution was purified by pre-HPLC to afford the compound (4 mg, 42% yield). MS (ESI): m / z found [M+H] + =1252.85.

[0650] Example 55:

[0651] Synthesis of compound 55:

[0652] Under ice, 50-1 (7.2 mg, 0.0118 mmol) was dissolved in 2 ml of ultra-dry DMF. 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (7.8 mg, 0.0134 mmol) and 1-hydroxybenzotriazole monohydrate (2 mg, 0.0134 mmol) were added and reacted for ten minutes. 11 (5 mg, 0.0118 mmol) and N,N-diisopropylethylamine (1.5 mg, 0.0224 mmol) were then added and reacted for ten minutes. The mixture was then warmed to room temperature and allowed to react for one hour. The reaction was complete after LC-MS monitoring. The reaction solution was purified by Pre-HPLC to give compound 55 (2.22 mg, 19%). MS (ESI): m / z found [M+H] + =1024.60.

[0653] Example 56:

[0654] Synthesis route:

[0655] Synthesis of compound 56:

[0656] Under ice, 56-1 (5.9 mg, 0.0071 mmol) was dissolved in 2 mL of ultra-dry DMF. 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (3.8 mg, 0.0066 mmol) and 1-hydroxybenzotriazole monohydrate (1.0 mg, 0.0066 mmol) were added and reacted for 10 minutes. Compound 11 (3.0 mg, 0.0071 mmol) and N,N-diisopropylethylamine (1.4 mg, 0.0135 mmol) were then added and reacted for 10 minutes. The mixture was then warmed to room temperature for one hour. LC-MS monitored the reaction to determine completion. The reaction solution was purified by Pre-HPLC to yield compound 56 (0.43 mg, 5%). MS (ESI): m / z found [M+H] + =1246.80.

[0657] Example 57:

[0658] Synthesis route:

[0659] Synthesis of intermediate 57-2:

[0660] Dissolve 1-(9H-fluoren-9-ylmethyl)-5,8,11,14,17,20-hexaoxa-2-azatricosanedioate (1.4 g, 1.73 mmol) in DMF (10 mL). Add piperidine (2.5 ml) and stir at room temperature for 3 h. LC-MS monitoring indicated that the reaction of 57-1 was complete. Diethyl ether was added to the reaction solution at 0°C, causing a white solid to precipitate. After standing at room temperature for 2 h, the white solid 57-2 (800 mg, 95%) was obtained by filtration.

[0661] Synthesis of intermediate 57-4:

[0662] 2-Chloropyrimidine-5-carboxylic acid (1 g, 6.3 mmol) and MeSNa (860 mg, 12.6 mmol) were dissolved in MeOH (200 ml), and KCO (2.7 g, 12.6 mmol) was added. The mixture was allowed to react overnight at room temperature. The reaction solution was concentrated, H O (50 ml) was added, and the mixture was extracted with EA (30 ml x 3). 2M HCl was added dropwise to the aqueous phase. A solid precipitated, and the mixture was allowed to stand for 1 hour before being filtered and lyophilized to obtain 57-4 (800 mg, 75%) as a white solid. MS (ESI): m / z found [M+H] + =171.2.

[0663] Synthesis of intermediate 57-5:

[0664] 57-4 (150 mg, 0.881 mmol) and N-hydroxysuccinimide (107.2 mg, 0.925 mmol) were dissolved in DCM (5 ml). 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (177.3 mg, 0.925 mmol) was added and stirred at room temperature for 3 h. LC-MS monitoring showed that the reaction of the starting material 57-4 was complete and it was used directly in the next step. MS (ESI): m / z found [M+H] + =268.2.

[0665] Synthesis of intermediate 57-6:

[0666] Compound 57-2 (311.2 mg, 0.881 mmol) was added to the reaction mixture of step 57-5, followed by triethylamine (122 μL, 0.881 mmol) and allowed to react at room temperature for 5 h. Rapid reaction chromatography was performed on a C-18 reverse phase column using 26% ACN in H₂O to afford a white solid (400 mg, 89%). MS (ESI): m / z found [M+H] + =506.2.2.

[0667] Synthesis of intermediate 57-7:

[0668] Compound 57-6 (400 mg, 0.79 mmol) was dissolved in DCM (5 ml), and 85% m-chloroperbenzoic acid (402 mg, 1.97 mmol) was added. The mixture was allowed to react at room temperature for 12 h. The mixture was then flash chromatographed on a C-18 reverse phase column using 35% ACN in H2O. After concentration, a colorless oily liquid (380 mg, 89%) was obtained. MS (ESI): m / z found [M+H] + =538.2.

[0669] Synthesis of intermediate 57-9:

[0670] 57-8 (14.4 mg, 0.032 mmol) was dissolved in DMF (3 ml), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (14 mg, 0.038 mmol) and 1-hydroxybenzotriazole (5 mg, 0.038 mmol) were added. The mixture was stirred at room temperature for 20 min. N,N-diisopropylethylamine (1 μL, 0.064 mmol) and compound 6 (20 mg, 0.032 mmol) were added, and the mixture was stirred at room temperature for 12 h. 30 ml of H2O was added to the reaction mixture, and the mixture was extracted with DCM (15 ml x 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography with MeOH in DCM (10%) to obtain a white solid (30 mg, 90%). MS (ESI): m / z found [M+H]+ =1024.55.

[0671] Synthesis of intermediate 57-10:

[0672] Compound 57-9 (30 mg, 0.029 mmol) was dissolved in DCM (5 ml), trifluoroacetic acid (1 ml) was added, and the mixture was stirred at room temperature for 2 h. The reaction solution was concentrated, dissolved in 2 ml of MeOH, and purified by flash chromatography on a C-18 reversed phase column using 60% ACN in H2O to afford a white solid (12 mg, 44%). MS (ESI): m / z found [M+H] + =924.2.

[0673] Synthesis of compound 57:

[0674] Compound 57-7 (5 mg, 0.0053 mmol) was dissolved in DMF (3 ml), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (2.5 mg, 0.0063 mmol) was added. The mixture was stirred at room temperature for 20 min. Compound 57-10 (3 mg, 0.0053 mmol) and N,N-diisopropylethylamine (1.35 μL, 0.0076 mmol) were added, and the mixture was stirred at room temperature for 3 h. The reaction mixture was purified by HPLC and concentrated to give the product as a light yellow solid (2 mg, 26%). MS (ESI): m / z found [M+H] + =1443.8.

[0675] Example 58:

[0676] Synthesis route:

[0677] 58-1 (5 mg, 0.0058 mmol) was dissolved in DMF (3 ml), and compound 6 (3.5 mg, 0.0058 mmol), DIEA (1 μL, 0.0058 mmol), HOBT (1.5 mg, 0.0117 mmol), and pyridine (2.31 μL, 0.029 mmol) were added and stirred at room temperature for 12 h. HPLC preparative purification with ACN in H2O (1‰ TFA) 5-55% for 30 min afforded a yellow solid (2.1 mg, 27%). MS (ESI): m / z found [M+H] + =1321.7.

[0678] Example 59:

[0679] Synthesis route:

[0680] Synthesis of intermediate 59-2:

[0681] 57-7 (40 mg, 0.074 mmol) and N-hydroxysuccinimide (9.5 mg, 0.082 mmol) were dissolved in DCM (5 ml), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (16 mg, 0.082 mmol) was added, followed by stirring at room temperature for 3 h. LC-MS monitored the reaction for completion. VAL-CIT-PAB (59-1, 28 mg, 0.082 mmol) and triethylamine (11.3 μL) were added, and the mixture was stirred at room temperature for 2 h. The reaction solution was concentrated, dissolved in 5 ml of MeOH, and purified by flash column chromatography (ACN in H2O 78%). MS (ESI): m / z found [M+H] + =899.2.

[0682] Synthesis of intermediate 59-3:

[0683] 59-2 (52 mg, 0.058 mmol) and di(p-nitrobenzene) carbonate (35 mg, 0.116 mmol) were dissolved in DMF (2.5 ml), and N,N-diisopropylethylamine (14.3 μL, 0.087 mmol) was added and allowed to react at room temperature for 18 h. 50 ml of H₂O was added to the reaction solution, and the mixture was extracted with DCM (20 ml x 3). The organic phases were combined, washed three times with water, and then concentrated. The combined organic phases were purified by flash column chromatography using MeOH in DCM (5%) to obtain a white solid (20 mg, 32%). MS (ESI): m / z found [M+1] + =1064.3.

[0684] Synthesis of compound 59:

[0685] 59-3 (20 mg, 0.019 mmol) was dissolved in DMF (3 mL), and 6 (11 mg, 0.019 mmol) and N,N-diisopropylethylamine (12 μL, 0.075 mmol) were added. The mixture was stirred at room temperature for 3 h. Purification by flash column chromatography using MeOH in DCM (8%) afforded 5 mg of the crude product as a white solid. HPLC preparation using ACN in H₂O (1‰ TFA) at 5-55% yielded 0.84 mg (3%) of the product as a pale yellow solid. MS (ESI): m / z found [M / 2+1] + =765.15.

[0686] Example 60:

[0687] Synthesis route:

[0688] Synthesis of intermediate 60-2:

[0689] 15-Azido-4,7,10,13-tetraoxopentadecanoic acid (76 mg, 0.26 mmol) was dissolved in DMF (5 ml), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (110 mg, 0.34 mmol) was added. The mixture was stirred at room temperature for 20 min. VAL-CIT-PAB (100 mg, 0.26 mmol) and DIEA (54 μL, 0.32 mmol) were added, and the mixture was stirred at room temperature for 12 h. The reaction mixture was purified by flash column chromatography on a C-18 reverse phase column using ACN in H2O (75%) to obtain the product as a white solid (120 mg, 75%). MS (ESI): m / z found [M+H] + =653.4.

[0690] Synthesis of intermediate 60-3:

[0691] Dissolve 60-2 (60 mg, 0.092 mmol) and di(p-nitrobenzene) carbonate (97 mg, 0.322 mmol) in DMF (2.5 ml) and add N,N-diisopropylethylamine (35 μL, 0.211 mmol). Heat to 50°C and react for 12 h. Add 30 ml of H2O to the reaction solution, extract with DCM (10 ml x 3), combine the organic phases, dry over anhydrous sodium sulfate, and concentrate. After concentration, column chromatography with MeOH in DCM (8%) was performed to obtain the product as a white solid (40 mg, 75.3%). MS (ESI): m / z found [M+H] + =818.4.

[0692] Synthesis of intermediate 60-4:

[0693] 60-3 (20 mg, 0.024 mmol) was dissolved in DMF (2.5 ml), and 6 (15 mg, 0.024 mmol) and N,N-diisopropylethylamine (16 μl, 0.096 mmol) were added. The mixture was reacted at room temperature for 3 h. ACN in H2O 5-55% (1‰ TFA) was prepared by HPLC for 30 min to give the product as a white solid (5 mg, 16%). MS (ESI): m / z found [M+H] + =1284.55.

[0694] Synthesis of compound 60:

[0695] Compounds 60-4 (3.8 mg, 0.0076 mmol) and 60-5 (5 mg, 0.0038 mmol) were dissolved in THF (0.5 ml) and EtOH (1.5 ml). Copper sulfate pentahydrate (1 mg, 0.0038 mmol) and L-ascorbic acid (0.7 mg, 0.0038 mmol) were added. The mixture was stirred at room temperature under nitrogen for 3 h and purified by HPLC using ACN in H2O (1‰ TFA) 5-55% for 30 min. The product was obtained as a light yellow solid (1.8 mg, 26%). MS (ESI): m / z found [M+H] + =1770.75.

[0696] Example 61:

[0697] Synthesis route:

[0698] Synthesis of intermediate 61-2:

[0699] Dissolve 57-7 (100 mg, 0.186 mmol) in DMF (3 mL), add 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (35 mg, 0.186 mmol), and stir at room temperature for 30 min. Then add DIEA (38 μL, 0.459 mmol) and 61-1 (80 mg, 0.186 mmol). React at room temperature for 5 h. Flash chromatography is performed on a C-18 reverse phase column using 38% ACN in H2O. Concentrate to obtain a white solid (90 mg, 51%). MS (ESI): m / z found [M+23] + =965.2.

[0700] Synthesis of compound 61:

[0701] Compound 61-2 (5.2 mg, 0.0055 mmol) was dissolved in DMF (2 ml), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (2.1 mg, 0.0055 mmol) and HOBT (0.7 mg, 0.0055 mmol) were added. The mixture was stirred at room temperature for 20 min. 11 (2.34 mg, 0.0055 mmol) and DIEA (1.82 μl, 0.011 mmol) were added, and the mixture was stirred at room temperature for 5 h. HPLC was used to prepare the mixture, 5%-50% (30 min), ACN in H2O (0.1% TFA), to give a yellow solid (2.4 mg, 32%). MS (ESI): m / z found [M+H] + =1350.7.

[0702] Example 62:

[0703] 61-2 (10 mg, 0.011 mmol) was dissolved in DMF (1.5 ml), and HATU (5 mg, 0.013 mmol), DIEA (2.2 μL, 0.013 mmol), and 12 (5 mg, 0.011 mmol) were added. The reaction was allowed to react at room temperature for 3 hours. After completion of the reaction as monitored by LC-MS, the product was purified by HPLC and lyophilized to give a yellow solid (0.50 mg, 3.4%). MS (ESI): m / z found [M / 2+1] + =684.35.

[0704] Example 63:

[0705] Compound 61-2 (11 mg, 0.012 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (4.1 mg, 0.022 mmol), compound 13 (5 mg, 0.012 mmol), and 2,4,6-trimethylpyridine (8.6 μL, 0.065 mmol) were added and the mixture was allowed to react at room temperature for 2 hours. LC-MS monitored the reaction completion. The reaction solution was purified by pre-HPLC to give compound 63 (0.7 mg, 4% yield). MS (ESI): m / z found [M+H] + =1376.80.

[0706] Example 64:

[0707] Compound 61-2 (11 mg, 0.0113 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (4.0 mg, 0.0203 mmol), compound 14 (5 mg, 0.0113 mmol), and 2,4,6-trimethylpyridine (8.0 μL, 0.0610 mmol) were added and the mixture was allowed to react at room temperature for 2 hours. LCMS monitored the reaction for completion. The reaction solution was purified by pre-HPLC to afford compound 64 (5.85 mg, 4% yield). MS (ESI): m / z found [M+H] + =1366.70.

[0708] Example 65:

[0709] Compound 61-2 (11 mg, 0.0116 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (4.0 mg, 0.0209 mmol), compound 15 (5.0 mg, 0.0116 mmol), and 2,4,6-trimethylpyridine (8.3 μL, 0.0610 mmol) were added and the mixture was allowed to react at room temperature for 2 hours. LCMS monitored the reaction to completion. The reaction solution was purified by pre-HPLC to give compound 65 (3.76 mg, 24% yield). MS (ESI): m / z found [M+H] + =1354.98.

[0710] Example 66:

[0711] Compound 61-2 (13 mg, 0.0134 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (4.6 mg, 0.0134 mmol), compound 24 (8 mg, 0.0134 mmol), and 2,4,6-trimethylpyridine (9.6 μL, 0.0724 mmol) were added and the mixture was allowed to react at room temperature for 2 hours. LCMS monitored the reaction to completion. The reaction solution was purified by pre-HPLC to give compound 66 (0.9 mg, 4% yield). MS (ESI): m / z found [M / 2+H] + =761.25.

[0712] Example 67:

[0713] 61-2 (20 mg, 0.021 mmol) was dissolved in DMF (3 ml), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (11.3 mg, 0.030 mmol) and 1-hydroxybenzotriazole (4.02 mg, 0.030 mmol) were added. The mixture was stirred at room temperature for 20 min, and 47 (9 mg, 0.021 mmol) and N,N-diisopropylethylamine (6.7 μL) were added. The mixture was purified by HPLC using ACN in H2O (1‰ HCOOH) 5-55% for 30 min. The product was obtained as a white solid (2 mg, 7%). MS (ESI): m / z found [M+H] + =1348.76.

[0714] Example 68:

[0715] Synthesis route:

[0716] Synthesis of intermediate 68-2:

[0717] Compound 68-1 (312 mg, 1.18 mmol) was added to the reaction mixture of 57-5 (200 mg, 1.18 mmol) from the previous step, followed by triethylamine (163 μL, 1.18 mmol) and allowed to react at room temperature for 5 h. Rapid reaction chromatography was performed on a C-18 reverse phase column using 26% ACN in H2O to yield a colorless oil (320 mg, 65%). MS (ESI): m / z found [M+H] + =418.2.

[0718] Synthesis of intermediate 68-3:

[0719] Compound 68-2 (320 mg, 0.767 mmol) was dissolved in DCM (5 mL), and 85% m-chloroperbenzoic acid (390 mg, 2.26 mmol) was added. The mixture was allowed to react at room temperature for 12 h. The mixture was then flash chromatographed on a C-18 reverse phase column using 35% ACN in H2O. After concentration, a colorless oily liquid (180 mg, 52.2%) was obtained. MS (ESI): m / z found [M+H] + =450.25.

[0720] Synthesis of intermediate 68-4:

[0721] Dissolve 68-3 (50 mg) in DMF (3 mL), add 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (42 mg), and stir at room temperature for 30 min. Add DIEA (46 μL) and 61-1 (48 mg). React at room temperature for 5 h. Purify by reverse-phase column chromatography (ACN in H2O 38%) and concentrate to obtain a white solid (50 mg). MS (ESI): m / z found [MH] - =853.2.

[0722] Synthesis of compound 68:

[0723] Compound 68-3 (10.1 mg, 0.0118 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (7.8 mg, 0.0135 mmol) and 1-hydroxybenzotriazole (1.9 mg, 0.0135 mmol) were added and allowed to react for 10 minutes. Compound 11 (5 mg, 0.0118 mmol) and N,N-diisopropylethylamine (3.9 μL, 0.0224 mmol) were then added and the reaction was maintained at room temperature for 2 hours. The reaction was complete as monitored by LCMS. The reaction solution was purified by pre-HPLC to obtain the compound (2.39 mg, 16% yield). MS (ESI): m / z found [M+H] + =1262.70.

[0724] Example 69:

[0725] Synthesis route:

[0726] Synthesis of intermediate 69-2:

[0727] Compound 69-1 (114.5 mg, 0.646 mmol) was added to the reaction mixture of 57-5, followed by triethylamine (90 μL, 0.646 mmol) and the mixture was allowed to react at room temperature for 5 h. Rapid reaction chromatography was performed on a C-18 reverse phase column using 28% ACN in H2O to obtain a colorless oil (100 mg, 58%). MS (ESI): m / z found [M+H] + =268.0.

[0728] Synthesis of intermediate 69-3:

[0729] Compound 69-2 (100 mg, 0.375 mmol) was dissolved in DCM (3 mL), and 85% m-chloroperbenzoic acid (154 mg, 0.936 mmol) was added. The mixture was allowed to react at room temperature for 12 h. The mixture was then subjected to rapid column chromatography on a C-18 reverse phase column using 18% ACN in H2O. After concentration, a colorless oily liquid (35 mg, 25.8%) was obtained. MS (ESI): m / z found [M+H] + =362.0.

[0730] Synthesis of intermediate 69-4:

[0731] Dissolve 69-3 (35 mg) in DMF (3 mL) and add 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (36 mg). Stir at room temperature for 30 min. Add DIEA (40 μL) and 61-1 (41 mg). React at room temperature for 5 h. Purify by HPLC (ACN in H2O 38%) and lyophilize to obtain the product. MS (ESI): m / z found [MH] - =765.3.

[0732] Synthesis of compound 69:

[0733] Compound 69-4 (9.1 mg, 0.0118 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (7.8 mg, 0.0135 mmol) and 1-hydroxybenzotriazole (1.9 mg, 0.0135 mmol) were added and allowed to react for 10 minutes. Compound 11 (5 mg, 0.0118 mmol) and N,N-diisopropylethylamine (3.9 μL, 0.0224 mmol) were then added and the reaction was maintained at room temperature for 2 hours. The reaction was complete as monitored by LCMS. The reaction solution was purified by pre-HPLC to obtain the compound (5.26 mg, 37% yield). MS (ESI): m / z found [M+H] + =1174.83.

[0734] Example 70:

[0735] The synthesis steps were the same as in Example 69 except that the corresponding starting materials were replaced to give a yellow solid product (5.26 mg, 37% yield). LC-MS (ESI): m / z found [M+H] + =1218.75.

[0736] Example 71:

[0737] Synthesis route:

[0738] Synthesis of intermediate 71-3:

[0739] 71-1 (200 mg, 0.876 mmol) and 71-2 (180 mg, 0.876 mmol) were dissolved in DMF, and TEA (0.66 mL), CuI (16.8 mg, 0.088 mmol), and PdCl2(PPh3)2 (61.8 mg, 0.088 mmol) were added sequentially. The mixture was heated to 95°C under N2 protection and reacted for 6 h. LC-MS monitored the reaction completion. The mixture was extracted with EA / H2O, washed with saturated NaCl, dried over anhydrous Na2SO4, filtered, and dried by spin drying. The product was purified by normal phase column chromatography using 8% EA:PE and dried to give a yellow solid (205 mg, 66.4%). LC-MS (ESI): m / z found [M+H] + =353.0.

[0740] Synthesis of intermediate 71-4:

[0741] 71-3 (91 mg, 0.258 mmol) was dissolved in DCM (3 mL) and TFA (1 mL) was added dropwise. The mixture was allowed to react at room temperature for 1 h. LC-MS confirmed the reaction was complete. The solvent was then removed by distillation under reduced pressure to give a yellow solid product (90.6 mg, 100%). LC-MS (ESI): m / z found [M+H] + =297.0.

[0742] Synthesis of intermediate 71-5:

[0743] Compound 71-4 (90.6 mg, 0.258 mmol) was dissolved in DCM (5 mL), and m-chloroperbenzoic acid (111.4 mg, 0.645 mmol) was added. The mixture was allowed to react at room temperature for 12 h. The mixture was then flash chromatographed on a C-18 reverse phase column using 16% ACN in H2O. After concentration, a white solid (49 mg, 48.8%) was obtained. LC-MS (ESI): m / z found [M+H] + =329.0.

[0744] Synthesis of intermediate 71-6:

[0745] 71-5 (25 mg, 0.076 mmol) was dissolved in MeCN, HOSU and DCC were added at 0°C, and the mixture was allowed to react at room temperature for 2 h. LC-MS monitored the reaction completion, and the white insoluble solid was removed by filtration. The filtrate was concentrated under reduced pressure to obtain a white solid product (32.3 mg, 99.9%). LC-MS (ESI): m / z found [M+H] + =426.0.

[0746] Synthesis of intermediate 71-7:

[0747] 71-6 (32.3 mg, 0.076 mmol) was dissolved in DMF (2 mL), and 61-1 (32.2 mg, 0.076 mmol) and DIEA (25 μL, 0.152 mmol) were added sequentially and reacted at room temperature for 2 h. After the reaction was complete, the product was purified by reverse-phase column chromatography and lyophilized to obtain the product (49.2 mg, 8.2%). LC-MS (ESI): m / z found [MH] - =732.25.

[0748] Synthesis of compound 71:

[0749] Compound 71-7 (8.7 mg, 0.0118 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (7.8 mg, 0.0135 mmol) and 1-hydroxybenzotriazole (1.9 mg, 0.0135 mmol) were added and allowed to react for 10 minutes. Compound 11 (5 mg, 0.0118 mmol) and N,N-diisopropylethylamine (3.9 μL, 0.0224 mmol) were then added and the reaction was maintained at room temperature for 2 hours. The reaction was complete as monitored by LCMS. The reaction solution was purified by pre-HPLC to obtain the compound (2.61 mg, 19% yield). MS (ESI): m / z found [M+H] + =1141.60.

[0750] Example 72:

[0751] Synthesis route:

[0752] Synthesis of intermediate 72-3:

[0753] On ice, compound N-2-Fluorenylmethyloxycarbonyl-L-2,4-diaminobutyric acid (72-1, 334 mg, 0.981 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide. Methyl-PEG8-NHS ester (72-2, 500 mg, 0.981 mmol) and N,N-diisopropylethylamine (267 μL, 1.530 mmol) were added and the mixture was allowed to react at room temperature for 2 hours. LCMS monitored the reaction completion. The reaction solution was purified by reverse-phase column chromatography and lyophilized to obtain the compound (605 mg, 84% yield). MS (ESI): m / z found [M+H] + =735.50.

[0754] Synthesis of intermediate 72-4:

[0755] Compound 72-3 (100 mg, 0.136 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. N-hydroxysuccinimide (20 mg, 0.163 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (32 mg, 0.163 mmol) were added and allowed to react for 1.5 hours. Compound 61-1 (58 mg, 0.136 mmol) and N,N-diisopropylethylamine (24 μL, 0.136 mmol) were then added and the reaction was maintained at room temperature for 2 hours. LCMS monitored the reaction completion. The reaction solution was purified by reverse phase column chromatography to obtain the compound (65 mg, 42% yield). MS (ESI): m / z found [M+H] + =1140.80.

[0756] Synthesis of intermediate 72-5:

[0757] Compound 72-4 (485 mg, 0.426 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide at room temperature. Piperidine (421 μL, 4.26 mmol) was added and the atmosphere was purged with nitrogen three times. The reaction was allowed to react at room temperature for 3 hours. LC-MS monitored the reaction to completion. The reaction solution was purified by reverse-phase column chromatography to obtain the compound (1.5 mg, 99% yield). MS (ESI): m / z found [M+H] + =918.70.

[0758] Synthesis of intermediate 72-6:

[0759] Compound 72-5 (37.8 mg, 0.0412 mmol) was dissolved in 3 ml of dry N,N-dimethylformamide under ice. Maleimidoacetic acid succinimidyl ester (10.4 mg, 0.0412 mmol) and N,N-diisopropylethylamine (6.8 μL, 0.0412 mmol) were added and the mixture was allowed to react at room temperature for 3 hours. LCMS monitored the reaction to completion. The reaction solution was purified by pre-HPLC to obtain the compound (17 mg, 39% yield). MS (ESI): m / z found [M+H] + =735.50.

[0760] Synthesis of compound 72:

[0761] Compound 72-6 (12.4 mg, 0.0118 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (7.8 mg, 0.0135 mmol) and 1-hydroxybenzotriazole (1.9 mg, 0.0135 mmol) were added and allowed to react for 10 minutes. Compound 11 (5 mg, 0.0118 mmol) and N,N-diisopropylethylamine (3.9 μL, 0.0224 mmol) were then added and the reaction was maintained at room temperature for 2 hours. LCMS monitored the reaction completion. The reaction solution was purified by pre-HPLC to yield 2.41 mg of compound (14% yield). LC-MS (ESI): m / z found [M+H] + =1462.90.

[0762] Example 73:

[0763] Synthesis route:

[0764] Synthesis of intermediate 73-3:

[0765] 73-1 (410 mg, 1 mmol) was dissolved in DMF, and DIEA (0.188 mL, 1.14 mmol) and HATU (0.433 g, 1.14 mmol) were added sequentially. The mixture was allowed to react at room temperature for 30 minutes. TLC analysis indicated the formation of an active ester intermediate. 73-2 (0.136 mL, 1 mmol) was added and allowed to react at room temperature for 2 hours. After the reaction was complete, the mixture was slurried in H2O / DMF, filtered, and the filter cake was lyophilized to obtain the product (501 mg, 95.8%). LC-MS (ESI): m / z found [M+H] + =524.45.

[0766] Synthesis of intermediate 73-4:

[0767] 73-3 (501 mg, 0.958 mmol) was dissolved in DCM (10 mL), TFA was added dropwise at 0°C, and the reaction was then allowed to proceed to room temperature. The mixture was concentrated under reduced pressure, dissolved in a small amount of DMF, and purified on a reverse phase column with 39% MeCN:H2O. The product was lyophilized to obtain a white solid (370 mg, 82.6%). LC-MS showed [M+H] + =468.2.

[0768] Synthesis of intermediate 73-5:

[0769] 73-4 (100 mg, 0.214 mmol) was dissolved in DMF, and Cu(OAc)2 (2 mg), Pb(OAc)4 (110 mg, 0.338 mmol), and CH3COOH (28.2 μL) were added sequentially. The mixture was heated to 60°C under N2 protection and reacted for 4 h. Some of the remaining raw material was extracted with EA / H2O, washed with saturated NaCl, dried over anhydrous Na2SO4, filtered, and dried by spin drying. The product (65.8 mg, 63.9%) was obtained by filtration. LC-MS showed [M+H] + =482.2.

[0770] Synthesis of intermediate 73-6:

[0771] 73-5 (646 mg, 1.34 mmol) was dissolved in DCM (14 mL), PPTS (700 mg, 2.81 mmol) was added, and the mixture was refluxed at 40°C overnight. Purification by normal phase column chromatography and concentration under reduced pressure gave a white solid (351 mg, 44.6%). LC-MS showed [M+Na] + =610.2.

[0772] Synthesis of intermediate 73-7:

[0773] Compound 73-6 (100 mg, 0.170 mmol) was dissolved in 5 ml of dry ethyl acetate and 10 ml of dry ethanol at room temperature. An appropriate amount of palladium-carbon was added, and the hydrogen atmosphere was replaced three times. The reaction was then maintained at room temperature for 4 hours. LCMS confirmed the reaction was complete. The reaction solution was filtered through celite, and the filtrate was concentrated and purified by reverse-phase column chromatography to afford the compound (25 mg, 30% yield). MS (ESI): m / z found [M+H] + =496.20.

[0774] Synthesis of intermediate 73-8:

[0775] Compound 73-7 (5.9 mg, 0.0118 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide at room temperature. O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (7.8 mg, 0.0135 mmol), compound 11 (5 mg, 0.0118 mmol), and N,N-diisopropylethylamine (3.9 μL, 0.0224 mmol) were added and the mixture was allowed to react at room temperature for 2 hours. LCMS monitored the reaction to completion. The reaction solution was purified by pre-HPLC to obtain the compound (2.3 mg, 22% yield). LC-MS (ESI): m / z found [M+H] + =905.55.

[0776] Synthesis of intermediate 73-9:

[0777] Compound 73-8 (2.3 mg, 0.0025 mmol) was dissolved in 3 ml of dry N,N-dimethylformamide at room temperature, and piperidine (2.5 μL, 0.025 mmol) was added. The mixture was allowed to react at room temperature for 1.5 hours. LC-MS monitored the reaction to completion. The reaction solution was purified by pre-HPLC to obtain compound 72-4 (1.5 mg, 87% yield). MS (ESI): m / z found [M+H] + =683.45.

[0778] Synthesis of compound 73:

[0779] Compound 73-9 (1.5 mg, 0.0022 mmol) was dissolved in 2 ml of dry N,N-dimethylformamide under ice. O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (1.0 mg, 0.0025 mmol), compound 57-7 (1.4 mg, 0.0022 mmol), and N,N-diisopropylethylamine (0.8 μL, 0.0044 mmol) were added and the mixture was allowed to react at room temperature for 2 hours. LCMS monitored the reaction to completion. The reaction solution was purified by pre-HPLC to obtain the compound (0.95 mg, 36% yield). MS (ESI): m / z found [M+H] + =1096.80.

[0780] Example 74:

[0781] Synthesis route:

[0782] Synthesis of intermediate 74-2:

[0783] 74-1 (500 mg, 3.96 mmol) and 70-2 (0.813 mg, 3.96 mmol) were dissolved in DMF (3 mL). TEA (3 mL), CuI (75 mg, 0.396 mmol), and PdCl2(PPh3)2 (278 mg, 0.396 mmol) were added sequentially. The mixture was heated to 95°C under N2 protection and reacted for 6 h. LC-MS monitored the reaction completion. The product was extracted with EA / H2O, washed with saturated NaCl, dried over anhydrous Na2SO4, filtered, and dried by spin drying. The product was purified by normal phase column chromatography and concentrated under reduced pressure to obtain a white solid (736 mg, 74.2%). LC-MS (ESI): m / z found [M+H] + =251.0.

[0784] Synthesis of intermediate 74-3:

[0785] 74-2 (0.375 g, 1.5 mmol) was dissolved in THF (8 mL), and H₂O (4 mL) and LiOH·H₂O (0.441 g, 10.5 mmol) were added sequentially. The mixture was allowed to react at room temperature for 4 h. After the reaction was complete, the reaction solution was washed twice with EA. The aqueous phase was adjusted to pH 2 with 1 M HCl aqueous solution. A large amount of white solid precipitated, which was filtered and lyophilized to obtain a white solid (273 mg, 77.1%). LC-MS (ESI): m / z found [M+H] + =237.0.

[0786] Synthesis of intermediate 74-4:

[0787] Compound 74-3 (540 mg, 0.488 mmol) was dissolved in DCM (40 mL), and m-chloroperbenzoic acid (766 mg, 4.437 mmol) was added. The mixture was reacted at room temperature for 12 h. After purification by reverse-phase column chromatography, the product was concentrated to give a white solid (483 mg, 83.9%). LC-MS (ESI): m / z found [M+H] + =269.2.

[0788] Synthesis of intermediate 74-5:

[0789] 74-4 (30 mg, 0.112 mmol) was dissolved in MeCN, HOSU and DCC were added at 0°C, and the mixture was allowed to react at room temperature for 2 h. LC-MS monitored the reaction completion, and the white insoluble solid was removed by filtration. The filtrate was concentrated under reduced pressure to obtain a white solid product (40 mg, 99%). LC-MS (ESI): m / z found [M+H] + =366.20.

[0790] Synthesis of intermediate 74-7:

[0791] Dissolve 74-6 (2 g, 9.23 mmol) in DCM (20 mL) and dissolve K2CO3 (3.8 g, 27.7 mmol) in H2O (8 mL). After cooling, add K2CO3 solution and (Boc)2O (2 g, 9.23 mmol). Allow to react at room temperature for 2 h. The reaction mixture is extracted with DCM / H2O, washed three times with saturated sodium chloride, concentrated, and purified using a normal phase column (17% EA:PE). Concentration under reduced pressure afforded the product as a pale yellow solid (980 mg, 37.9%). LC-MS (ESI): m / z found [MH] + =279.0.

[0792] Synthesis of intermediate 74-8:

[0793] 74-7 (980 mg, 3.5 mmol) was dissolved in EtOH (20 mL), and NaBH4 (200 mg, 5.28 mmol) was added. The mixture was reacted at room temperature under N2 for 2 h. The mixture was extracted with EA / H2O, washed with saturated NaCl, dried over anhydrous Na2SO4, filtered, and dried. The product was purified by normal phase column chromatography using 25% EA:PE and dried to give a light yellow solid (590 mg, 60%). LC-MS (ESI): m / z found [M-56] + =227.20.

[0794] Synthesis of intermediate 74-9:

[0795] 74-8 (590 mg, 2.09 mmol) was dissolved in MeOH (10 mL), and Pd / C was added. The mixture was reacted under H₂ for 4 h at room temperature. After the reaction was complete, the mixture was filtered, and the filtrate was dried and purified using a normal phase column (70% EA). The product was concentrated under reduced pressure to obtain a white solid (450 mg, 85%). LC-MS (ESI): m / z found [M-56] + =179.20.

[0796] Synthesis of intermediate 74-10:

[0797] Fmoc-VA (732 mg, 1.78 mmol) and HATU (814 mg, 2.136 mmol) were dissolved in DMF (4 mL) and stirred for 5 min. 74-9 (450 mg, 1.78 mmol) and DIEA (692 mg, 5.35 mmol) were added sequentially and allowed to react at room temperature. After the reaction was complete, the mixture was purified on a C-18 column (48% MeCN in H2O) and concentrated under reduced pressure to yield a white solid (1.05 g, 91%). LC-MS (ESI): m / z found [M-100] + =546.5.

[0798] Synthesis of intermediate 74-11:

[0799] 74-10 (200 mg, 0.31 mmol) was dissolved in DCM (2 mL), HCl / EA (0.39 mL) was added, and the mixture was allowed to react at room temperature for 4 h. After the reaction was complete, the mixture was concentrated under reduced pressure and dissolved in a small amount of DMF. The mixture was purified by reverse-phase column chromatography (50% ACN in H2O) and lyophilized to obtain a white solid (380 mg, 56%). LC-MS (ESI): m / z found [M+H] + =545.2.

[0800] Synthesis of intermediate 74-12:

[0801] 74-11 (380 mg, 0.698 mmol) was dissolved in DMF (2 mL), and m-PEG8-NHS ester (355.5 mg, 0.698 mmol) and DIEA (270 mg, 2.09 mmol) were added and reacted at room temperature. After the reaction was complete, the product was purified by reverse C-18 column (42% MeCN in H2O) and lyophilized to obtain a white solid (350 mg, 53.4%). LC-MS (ESI): m / z found [M+H] + =939.7.

[0802] Synthesis of intermediate 74-13:

[0803] Dissolve 74-12 (350 mg, 0.374 mmol) in DMF and stir for 5 minutes. Then add (PNP)2CO (226.7 mmol) and DIEA (48 mg, 0.374 mmol) sequentially and react at room temperature. After the reaction is complete, purify the mixture on a C-18 column (55% ACN in H2O) and lyophilize to obtain a white solid product. LC-MS (ESI): m / z found [M / 2+H] + =553.5.

[0804] Synthesis of intermediate 74-14:

[0805] 74-13 (50 mg, 0.045 mmol) and HOBt (6.75 mg, 0.050 mmol) were dissolved in DMF, and 11 (19.25 mg, 0.045 mmol) and DIEA (17.5 mg, 0.136 mmol) were added sequentially. The mixture was allowed to react at room temperature for 2 h. HPLC purification (10%-70% MeCN in H2O, 30 min) and lyophilization afforded the product as a yellow solid (35 mg, 56%). LC-MS (ESI): m / z found [M / 2+H] + =696.30.

[0806] Synthesis of intermediate 74-15:

[0807] 74-14 (35 mg, 0.025 mmol) was dissolved in DMF, diethylamine was added, and the mixture was allowed to react at room temperature for 2 h. Purification by HPLC (10%-60% ACN in H2O, 30 min) and lyophilization afforded a yellow solid (6 mg, 20.4%). LC-MS (ESI): m / z found [M+H] + =1168.80.

[0808] Synthesis of compound 74:

[0809] Dissolve 73-5 (2 mg, 0.005 mmol) in DMF, add 74-15 (3 mg, 0.003 mmol) and TEA (0.66 mg, 0.005 mmol), and react at room temperature for 2 h. Purify by HPLC and lyophilize to obtain a yellow solid (0.6 mg, 16.5%). LC-MS (ESI): m / z found [M / 2+H] + =710.35.

[0810] Example 75:

[0811] Synthesis route:

[0812] Synthesis of intermediate 75-1:

[0813] 68-3 (30 mg, 0.083 mmol) was dissolved in MeCN, HOSU and DCC were added at 0°C, and the mixture was allowed to react at room temperature for 2 h. LC-MS monitored the reaction completion. The white insoluble solid was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain a white solid product (38 mg, 99%). LC-MS (ESI): m / z found [M+H] + =459.1.

[0814] Synthesis of compound 75:

[0815] 75-1 (2 mg, 0.005 mmol) was dissolved in DMF, and 74-15 (3 mg, 0.003 mmol) and TEA (0.66 mg, 0.005 mmol) were added and reacted at room temperature for 2 h. Purification by HPLC and lyophilization afforded a yellow solid (0.63 mg, 16.2%). LC-MS (ESI): m / z found [M / 2+H] + =757.05.

[0816] Example 76:

[0817] Synthesis route:

[0818] Synthesis of intermediate 76-1:

[0819] 74-4 (43 mg, 0.177 mmol) was dissolved in DCM (5 mL), and HOSu (21.4 mg, 0.186 mmol) and EDCI (35.7 mg, 0.186 mmol) were added. The mixture was allowed to react at room temperature until the concentration of 74-4 ceased to decrease. 72-5 (162 mg, 0.177 mmol) and TEA (24 μL) were then added and the reaction continued at room temperature overnight. After the reaction was complete, the mixture was concentrated under reduced pressure, dissolved in MeOH, and prepared by HPLC to obtain a colorless oil (54.3 mg, 26.3%). LC-MS (ESI): m / z found [M-75] = 1092.65.

[0820] Synthesis of compound 76:

[0821] 76-1 (8.2 mg, 0.007 mmol) was dissolved in DMF, HATU (3.06 mg, 0.008 mmol) was added, and the mixture was allowed to react at room temperature for 30 min. 11 (3 mg, 0.007 mmol) and DIEA (1.3 μL, 0.008 mmol) were then added, and the reaction was continued at room temperature for 2 h. The reaction was monitored for completion by LC-MS, and the product was purified by HPLC (0.05% TFA-water) and lyophilized to obtain the product (3.23 mg, 29.3%). LC-MS (ESI): m / z found [M / 2+1] + =788.95.

[0822] Example 77:

[0823] Synthesis route:

[0824] Synthesis of intermediate 77-2:

[0825] 77-1 (300 mg, 0.6 mmol) was dissolved in DMF, and EDCI (126 mg, 0.66 mmol) and HOSU (76 mg, 0.66 mmol) were added. The mixture was allowed to react at room temperature for 2 h. DIEA (155 mg, 1.2 mmol) and 72-1 (204 mg, 0.6 mmol) were then added and allowed to react at room temperature for another 4 h. After completion of the reaction as monitored by LC-MS, the mixture was purified by reverse-phase C-18 column (50% ACN in H2O) and concentrated under reduced pressure to afford a colorless oil (193 mg, 39%). LC-MS (ESI): m / z found [M+H] + =823.6.

[0826] Synthesis of intermediate 77-3:

[0827] 77-2 (308 mg, 0.374 mmol) was dissolved in DMF, and DCC (85 mg, 0.412 mmol) and HOSU (47.4 mg, 0.412 mmol) were added. The mixture was allowed to react at room temperature for 2 h. DIEA (48 mg, 0.374 mmol) and 61-1 (158 mg, 0.374 mmol) were then added and allowed to react at room temperature for another 4 h. The mixture was purified by reverse-phase C-18 column purification (44% ACN in H2O) and concentrated under reduced pressure to afford the product (2225 mg, 49%). LC-MS (ESI): m / z found [M+H] + =1228.85.

[0828] Synthesis of intermediate 77-4:

[0829] 77-3 (225 mg, 0.183 mmol) was dissolved in DMF, diethylamine (50 μL) was added, and the mixture was allowed to react at room temperature for 2 h. Purification was performed on a reverse phase column (30% ACN in H2O) and concentrated under reduced pressure to obtain the product (115 mg, 62%). LC-MS (ESI): m / z found [M+H] + =1006.75.

[0830] Synthesis of intermediate 77-5:

[0831] Dissolve 77-4 (115 mg, 0.114 mmol) in DMF, add 74-5 (62 mg, 0.171 mmol) and DIEA (44 mg, 0.342 mmol), and react at room temperature for 2 h. Purify with a reverse phase column and lyophilize to obtain the product (100 mg, 70%). LC-MS (ESI): m / z found [M+H] + =1257.00.

[0832] Synthesis of compound 77:

[0833] 77-5 (30 mg, 0.024 mmol) and HATU (10 mg, 0.026 mmol) were dissolved in DMF, and 11 (10 mg, 0.024 mmol) and DIEA (9.1 mg, 0.072 mmol) were added and reacted at room temperature for 2 h. Purification by pre-HPLC and lyophilization afforded a yellow solid (11 mg, 28%). LC-MS (ESI): m / z found [M / 2+H] + =833.05.

[0834] Examples 78-80:

[0835] Referring to the synthesis method of Example 77, replacing the corresponding reaction raw materials, Examples 78-80 were obtained.

[0836] Table 1 Structural formula and MS (ESI) of Examples 78-80

[0837] Examples 81-82:

[0838] Referring to the synthesis methods of Example 72 and Example 76, the corresponding reaction raw materials were replaced to obtain Examples 81-82.

[0839] Table 2 Structural formula and MS (ESI) of Examples 81-82

[0840] Example 83:

[0841] Synthesis route:

[0842] Synthesis of intermediate 83-3:

[0843] 83-1 (100 mg, 0.326 mmol) was dissolved in THF (3 mL), and a catalytic amount of DMF was added. Oxalyl chloride (138 μL, 0.038 mmol) was added dropwise at 0°C under nitrogen. The reaction was then allowed to proceed at room temperature for 1 h, after which the solution turned clear from turbid. The product was then concentrated under reduced pressure. The acyl chloride intermediate was dissolved in DCM (2 mL), and TEA (453 μL, 3.26 mmol) and morpholine (143 μL, 1.632 mmol) were added at 0°C. The reaction was then allowed to proceed at room temperature for 2 h. The product was extracted with DCM / H₂O, washed with saturated NaCl, dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. Purification was then performed on a normal phase column (5.9% MeOH:DCM) to afford a light brown oil (141 mg, 97.3%). LC-MS (ESI): m / z found [M+H] + =445.0.

[0844] Synthesis of intermediate 83-4:

[0845] 83-3 (2.89 g, 6.5 mmol) was dissolved in EtOH (25 mL) and NaBH4 (1.967 g, 52 mmol) was added twice, with a 0.5 h interval between each addition. The reaction was allowed to react at room temperature overnight under nitrogen. The product was quenched with saturated aqueous NH4Cl solution, extracted with EA, dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and purified by normal phase column chromatography (10.3% MeOH:DCM). Concentration under reduced pressure gave a light yellow oil (1.25 g, 41.1%). LC-MS (ESI): m / z found [M+H] + =224.2.

[0846] Synthesis of intermediate 83-7:

[0847] 83-5 (200 mg, 1.142 mmol) and 83-6 (340.8 mg, 1.142 mmol) were dissolved in DMF (5 mL), and K2CO3 (316 mg, 2.284 mmol) was added. The mixture was refluxed at 100°C overnight. Extraction was performed with 1N dilute hydrochloric acid / DCM, and the mixture was dried over anhydrous Na2SO4. The mixture was filtered and concentrated under reduced pressure. Purification was performed on a normal phase column (15% EA:PE) and concentrated under reduced pressure to obtain a light yellow oil (234.7 mg, 68.2%). LC-MS (ESI): m / z found [M+H] + =302.2.

[0848] Synthesis of intermediate 83-8:

[0849] 83-7 (235 mg, 0.780 mmol) was dissolved in EtOH (8 mL). H2O (2 mL), Fe (174.3 mg, 3.12 mmol), and NH4Cl (66.8 mg, 1.248 mmol) were added sequentially, and the mixture was heated to 80°C and refluxed for 5 h. Filtered through Celite, the filtrate was concentrated under reduced pressure, and purified on a normal phase column (18.8% EA:PE). Concentration under reduced pressure afforded a light yellow oil (127.2 mg, 60.1%). LC-MS (ESI): m / z found [M+H] + =272.2.

[0850] Synthesis of intermediate 83-9:

[0851] 83-8 (23 mg, 0.085 mmol) was dissolved in 1 mL of AcOH, 2,3-dibromomaleic anhydride (21.7 mg, 0.085 mmol) was added, and the mixture was heated to 110°C and refluxed for 3 h. The mixture was evaporated under reduced pressure, purified by a normal phase column (15.8% EA:PE), and concentrated under reduced pressure to obtain a white solid (32.9 mg, 76.0%). LC-MS (ESI): m / z found [M+Na] + =526.8.

[0852] Synthesis of intermediate 83-10:

[0853] 83-9 (32.9 mg, 0.065 mmol) was dissolved in 2 mL of DCM. 83-4 (28.9 mg, 0.129 mmol) and DIEA (21.3 μL, 0.129 mmol) were slowly added at 0°C. The mixture was stirred at 0°C for 5 min and then allowed to react at room temperature for 2 h. The mixture was evaporated under reduced pressure, purified by a normal phase column, and concentrated under reduced pressure to yield an orange solid (16.3 mg, 31.6%). LC-MS (ESI): m / z found [M+H] + =794.0.

[0854] Synthesis of intermediate 83-12:

[0855] Compound 15-azido-4,7,10,13-tetraoxopentadecanoic acid (83-11, 50 mg, 0.0171 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. N-hydroxysuccinimide (26 mg, 0.257 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (50 mg, 0.257 mmol) were added and allowed to react for one hour. Compound 61-1 (109 mg, 0.257 mmol) and triethylamine (26 μL, 0.189 mmol) were then added and allowed to react at room temperature for 2 hours. The reaction was complete as monitored by LCMS. The reaction solution was purified by pre-HPLC to yield the compound (65 mg, 54%). LC-MS (ESI): m / z found [M+H] + =697.50.

[0856] Synthesis of intermediate 83-13:

[0857] Compound 83-12 (20 mg, 0.0287 mmol) was dissolved in 5 ml of dry N,N-dimethylformamide under ice. 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (12.4 mg, 0.0327 mmol) was added and allowed to react for 20 minutes. Compound 11 (12 mg, 0.0287 mmol) and N,N-diisopropylethylamine (9.7 μL, 0.0540 mmol) were then added and the reaction was maintained at room temperature for 2 hours. LC-MS monitored the reaction completion. The reaction solution was purified by pre-HPLC to obtain the compound (15 mg, 47% yield). LC-MS (ESI): m / z found [M+H] + =1104.07.

[0858] Synthesis of compound 83:

[0859] At room temperature, compound 83-13 (5 mg, 0.0045 mmol) was dissolved in 3 ml of dry tetrahydrofuran and 1 ml of dry ethanol. Compound 83-10 (3.6 mg, 0.0045 mmol), copper sulfate pentahydrate (1.12 mg, 0.0045 mmol), and vitamin C (0.79 mg, 0.0045 mmol) were added. The atmosphere was purged with nitrogen three times and the reaction was maintained at room temperature for 2 hours. LCMS monitored the reaction to completion. The reaction solution was dried and purified by pre-HPLC to obtain the compound (1.60 mg, 19% yield). LC-MS (ESI): m / z found [M / 2+H] + =949.00.

[0860] Examples 84-88:

[0861] Table 3 Structural formula and MS (ESI) of Examples 84-87

[0862] Example 89:

[0863] Synthesis route:

[0864] Synthesis of intermediate 89-3:

[0865] Compound 89-1 (200 mg, 0.3176 mmol) was dissolved in DCM (6.67 ml) at room temperature, and TFA (1.33 ml) and ethylene glycol (89-2, 0.333 ml) were added. The mixture was allowed to react at room temperature for 2 h. LC-MS confirmed the complete reaction of the starting material. The reaction solution was extracted with DCM / H₂O, and the organic phase was washed with saturated brine, dried over anhydrous Na₂SO₄, and concentrated. The product was then purified by normal phase column chromatography (methanol:dichloromethane = 13:87) to afford the product (130 mg, 65% yield). LC-MS (ESI): m / z found [M+H] + =632.5.

[0866] Synthesis of intermediate 89-4:

[0867] Compound 89-3 (125 mg, 0.198 mmol) was dissolved in DMF (2.5 ml) at room temperature, and DIEA (50 mg, 0.396 mmol) and di(p-nitrobenzene) carbonate (125 mg, 0.396 mmol) were added. The mixture was allowed to react at room temperature for 4 h. TLC and LC-MS confirmed that the starting material was no longer reactive. The reaction solution was extracted with DCM and water, and the organic phase was concentrated and purified by normal phase column chromatography (methanol:dichloromethane = 1:9) to obtain the product (70 mg, 44% yield). LC-MS (ESI): m / z found [M+H] + =797.5.

[0868] Synthesis of intermediate 89-5:

[0869] Compound 89-4 (70 mg, 0.0878 mmol) was dissolved in DMF (2 ml) at room temperature, and HOBT (13.5 mg, 0.0966 mmol) was added. The mixture was stirred for 5 min, followed by the addition of DIEA (22.7 mg, 0.1757 mmol) and 11 (38.8 mg, 0.0878 mmol). The mixture was allowed to react at room temperature for 2 h. LC-MS confirmed the absence of the starting material. The reaction solution was filtered and purified by a reverse phase C-18 column (55% MeCN in H2O). The product was concentrated to obtain 35 mg (37% yield). LC-MS (ESI): m / z found [M+H] + =1083.3.

[0870] Synthesis of intermediate 89-6:

[0871] Compound 89-5 (35 mg, 0.0323 mmol) was dissolved in DMF (2 ml) at room temperature, and piperidine (14 mg, 0.1615 mmol) was added. The mixture was reacted at room temperature for 2 h. LC-MS monitored the complete reaction of the starting material. The reaction solution was filtered and purified by Pre-HPLC (5%-50% ACN in H2O, 30 min, 0.1% formic acid system). The product was lyophilized to obtain 8.2 mg (29% yield). MS (ESI): m / z found [M+H] + =861.55.

[0872] Synthesis of compound 89:

[0873] Compound 57-7 (7.7 mg, 14.3 μmol) was dissolved in DMF (2 ml) at room temperature, and HATU (5.8 mg, 15.24 μmol) was added. The mixture was stirred for 5 min, followed by the addition of DIEA (3.7 mg, 28.6 μmol) and 89-6 (8.2 mg, 9.525 μmol). The mixture was allowed to react at room temperature for 2 h. LC-MS confirmed the absence of the starting material. The reaction solution was filtered and purified by Pre-HPLC (10%-55% ACN in H2O, 30 min, 0.1% formic acid system). The product was lyophilized to obtain 3.0 mg, 22% yield. MS (ESI): m / z found [M+H] + =1381.7.

[0874] Example 89:

[0875] Synthesis route:

[0876] Synthesis of intermediate 90-1:

[0877] At room temperature, 90-1 (465 mg, 0.7553 mmol) was dissolved in DMF (5 ml), and copper acetate (1 mg, 0.0038 mmol), acetic acid (104 mg, 1.737 mmol), and lead acetate (388 mg, 1.193 mmol) were added. The mixture was reacted at 60°C under nitrogen for 4 h. TLC confirmed the complete reaction of the starting material. The reaction solution was purified by reverse-phase C-18 column (50% MeCN in H2O) and concentrated to obtain the product (330 mg, 69% yield). LC-MS (ESI): m / z found [M+H] + =570.45.

[0878] Synthesis of intermediate 90-2:

[0879] Compound 90-1 (244 mg, 0.388 mmol) and TsOH (33 mg, 0.194 mmol) were dissolved in THF (3 ml) at room temperature. Benzyl (1s, 3s)-3-hydroxycyclobutane-1-carboxylate (160 mg, 0.777 mmol) was added and the mixture was allowed to react at room temperature for 2 h under N₂ protection. LC-MS confirmed the completion of the reaction. The reaction solution was extracted with DCM / H₂O, and the organic phase was washed with saturated brine, dried over anhydrous Na₂SO₄, concentrated, and purified by normal phase column chromatography (methanol:dichloromethane = 1:9) to afford product 1-3 (140 mg, 46.7% yield). MS (ESI): m / z found [M+H] + =776.5.

[0880] Synthesis of intermediate 90-3:

[0881] Compound 90-2 (140 mg, 0.18 mmol) was dissolved in EtOAc (2 ml) and EtOH (2 ml) at room temperature. Pd / C (0.096 mg, 0.009 mmol) was added to displace the hydrogen atmosphere, and the mixture was allowed to react at room temperature for 16 h. After LC-MS monitoring indicated that the starting material no longer reacted, the reaction solution was filtered and purified by reverse-phase C-18 column (47% MeCN in H2O). The product was concentrated to obtain 65 mg, 52.8% yield. MS (ESI): m / z found [M+H] + =686.5.

[0882] Synthesis of intermediate 90-4:

[0883] Compound 90-3 (65 mg, 95 μmol) was dissolved in DMF (2 ml) at room temperature, and HATU (39.6 mg, 104 μmol) was added. The mixture was stirred for 5 min, followed by the addition of DIEA (12.3 mg, 189.6 μmol) and 11 (40.3 mg, 95 μmol). The mixture was allowed to react at room temperature for 2 h. After LC-MS monitoring indicated that the starting material no longer reacted, the reaction solution was filtered and purified by reverse-phase C-18 column (55% MeCN in H2O). The product was concentrated to obtain the product (50 mg, 48% yield). MS (ESI): m / z found [M+H] + =1093.65.

[0884] Synthesis of intermediate 90-5:

[0885] At room temperature, compound 90-4 (50 mg, 0.0457 mmol) was dissolved in DMF (1.25 ml) and added

[0886] Piperidine (19.5 mg, 0.2285 mmol) was added and reacted at room temperature for 2 h. LC-MS monitored the complete reaction of the starting material. The reaction solution was filtered and purified by Pre-HPLC (5%-50% ACN in H2O, 30 min, 0.1% formic acid system). The product (13 mg, 32.6% yield) was lyophilized. MS (ESI): m / z found [M+H] + =871.55.

[0887] Synthesis of compound 90:

[0888] At room temperature, 57-7 (8.8 mg, 16.4 μmol) was dissolved in DMF (2 ml), HATU (6.8 mg, 17.9 μmol) was added, and the mixture was stirred for 5 min. DIEA (5.8 mg, 45 μmol) and 90-5 (13 mg, 15 μmol) were then added and reacted at room temperature for 2 h. LC-MS confirmed the absence of the starting material. The reaction solution was filtered and purified by Pre-HPLC (10%-50% ACN in H2O, 30 min, 0.1% formic acid system). The product was lyophilized to obtain 5.0 mg, 24% yield. MS (ESI): m / z found [M+H] + =1390.75.

[0889] Examples 91-96:

[0890] Synthesis route:

[0891] Synthesis of intermediate 91-2:

[0892] 91-1 (1 g, 1.7 mmol) was dissolved in DMF (8 mL), and EDCI (360 mg, 1.87 mmol) and HOSU (220 mg, 1.87 mmol) were added. The mixture was allowed to react at room temperature for 2 h. DIEA (320 mg, 2.55 mmol) and 72-1 (520 mg, 1.47 mmol) were then added and allowed to react at room temperature for another 4 h. After the reaction was complete, the mixture was purified by reverse phase column chromatography and concentrated under reduced pressure to obtain a colorless oil (980 mg, 63%). LC-MS (ESI): m / z found [M+H] + =911.7.

[0893] Except for replacing the corresponding reaction raw materials, Examples 91-96 were synthesized by referring to the synthesis method of Example 77 and Example 88. Their structural formulas and LC-MS (ESI) are shown in Table 4.

[0894] Table 4 Structural formula and MS (ESI) of Examples 91-96

[0895] Example 97:

[0896] Synthesis route:

[0897] Synthesis of intermediate 97-1:

[0898] Compound 90-3 (270 mg, 0.394 mmol) was dissolved in DMF, piperidine (167 mg, 1.968 mmol) was added, and the mixture was allowed to react at room temperature for 2 h. Purification was performed on a reverse phase column and concentration was performed under reduced pressure to obtain a white solid product (129 mg, 70%). LC-MS (ESI): m / z found [M+H] + =464.4.

[0899] Synthesis of intermediate 97-2:

[0900] 91-2 (253 mg, 0.278 mmol) was dissolved in DMF, followed by the addition of HATU (116 mg, 0.306 mmol) and DIEA (72 mg, 0.556 mmol). After stirring at room temperature for 30 min, compound 96-1 (129 mg, 0.278 mmol) was added and allowed to react at room temperature for another 2 h. Pre-HPLC preparation and concentration under reduced pressure afforded a colorless oil (120 mg, 31%). LC-MS (ESI): m / z found [M+H] + =1357.0.

[0901] Synthesis of intermediate 97-3:

[0902] Dissolve 97-2 (120 mg, 0.088 mmol) in DMF, add piperidine (38 mg, 0.442 mmol), and react at room temperature for 2 h. Pre-HPLC preparation and concentration under reduced pressure afforded a colorless oil (61 mg, 61%). LC-MS (ESI): m / z found [M+H] + =1134.85.

[0903] Synthesis of intermediate 97-4:

[0904] 97-3 (61 mg, 0.054 mmol) was dissolved in DMF, followed by the addition of 74-5 (21.6 mg, 0.059 mmol) and DIEA (20.85 mg, 0.161 mmol) at room temperature. After the reaction was complete, the mixture was purified by Pre-HPLC and concentrated under reduced pressure to obtain a light yellow oil (21 mg, 28%). LC-MS (ESI): m / z found [M / 2+H] + =693.3.

[0905] Synthesis of compound 97:

[0906] 97-4 (7 mg, 0.005 mmol) and HATU (2.1 mg, 0.005 mmol) were dissolved in DMF, DIEA (2 mg, 0.015 mmol) and 11 (2.36 mg, 0.005 mmol) were added, and the mixture was reacted at room temperature for 2 h. Pre-HPLC preparation and lyophilization gave a yellow solid (1.6 mg, 17.66%). LC-MS (ESI): m / z found [M / 2+H] + =897.15.

[0907] Examples 98-101:

[0908] Except for replacing the corresponding reaction raw materials, the synthesis method of Examples 98-101 was referred to that of Example 97, and their structural formulas and LC-MS (ESI) are shown in Table 5.

[0909] Table 5 Structural formula and LC-MS (ESI) of Examples 98-101

[0910] Example 102:

[0911] Synthesis route:

[0912] Synthesis of intermediate 102-1:

[0913] Dissolve Fmoc-VC (2.36 g, 4.756 mmol) and HATU (2.17 g, 5.707 mmol) in DMF (5 mL) and stir for 5 min. Then add 74-9 (1.2 g, 4.756 mmol) and DIEA (1.84 g, 14.268 mmol) sequentially. Allow to react at room temperature. After the reaction is complete, purify the mixture on a C-18 column (50% MeCN in H2O) and concentrate under reduced pressure to yield a white solid. LC-MS (ESI): m / z found [M+H] + =731.6.

[0914] Synthesis of intermediate 102-2:

[0915] 102-1 (710 mg, 0.972 mmol) was dissolved in DMF (5 mL), and HCl / EA (1.21 mL) was added and reacted at room temperature for 4 h. After the reaction was complete, the mixture was concentrated under reduced pressure to obtain a white solid (620 mg, 96%). LC-MS (ESI): m / z found [M+H] + =631.2.

[0916] Synthesis of intermediate 102-3:

[0917] 102-2 (1.8 g, 2.82 mmol) was dissolved in DMF (5 mL), and m-PEG8-NHS ester (1.7 g, 2.82 mmol) and DIEA (0.93 mL, 5.64 mmol) were added and reacted at room temperature. After the reaction was complete, the product was purified by reverse C-18 column and lyophilized to obtain a colorless oil (1.5 g, 51.9%). LC-MS (ESI): m / z found [M+H] + =1025.80.

[0918] Synthesis of intermediate 102-4:

[0919] 102-3 (1.5 g, 1.46 mmol) was dissolved in DMF, diethylamine (150 μL) was added, and the mixture was reacted at room temperature for 4 hours. The product was purified by reverse phase column chromatography (H2O:ACN=1:4) to obtain the product. LC-MS (ESI): m / z found [M+H] + =803.70.

[0920] Synthesis of intermediate 102-5:

[0921] 102-4 (560 mg, 0.7 mmol) was dissolved in DMF (4 mL), and maleimide-PEG2-succinimide ester (0.7 mmol) and DIEA (231 μL) were added. The reaction was stirred at room temperature for 3 hours. Purification was performed by reverse phase column chromatography (H2O:ACN=7:3) and concentration under reduced pressure to obtain the product (350 mg, 48.1%). LC-MS (ESI): m / z found [M+H] + =1042.75.

[0922] Synthesis of intermediate 102-6:

[0923] 102-5 (170 mg, 0.020 mmol) and (PNP)2CO (199 mg, 0.076 mmol) were dissolved in DMF (3 mL), and DIEA (51 μL) was added. The reaction was allowed to proceed at 40°C for 2 hours, then at room temperature overnight. The product was purified by reverse-phase column chromatography and lyophilized to obtain the product (90 mg, 45.9%).

[0924] Synthesis of compound 102:

[0925] 102-6 (30 mg, 0.022 mmol) was dissolved in DMF (2 ml), and HOBt (4 mg, 0.029 mmol), DIEA (7.3 μL, 0.044 mmol), and 11 (9 mg, 0.022 mmol) were added. The mixture was allowed to react at room temperature for 2.5 hours. After completion of the reaction as monitored by LC-MS, the product was purified by HPLC and lyophilized to give a yellow solid (1.94 mg, 6.5%). LC-MS (ESI): m / z found [M / 2+H] + =748.05.

[0926] Example 102:

[0927] Synthesis route:

[0928] Synthesis of intermediate 103-2:

[0929] 46-1 (500 mg, 2.016 mmol), N-tert-butyloxycarbonyl-L-alaninamide (103-1, 456 mg, 2.419 mmol), Pd2(dba)3 (93 mg, 0.1008 mmol), CsCO3 (1314 mg, 4.032 mmol), and XPhos (192 mg, 0.4032 mmol) were dissolved in toluene at room temperature. After nitrogen replacement, the temperature was raised to 90°C and the reaction was allowed to proceed overnight. TLC monitoring indicated the reaction was complete. The reaction solution was extracted with ethyl acetate and water. The organic phase was then washed with saturated aqueous NaCl solution, concentrated to dryness over anhydrous Na2SO4, and purified by normal phase column chromatography (petroleum ether:ethyl acetate = 77.7:22.3) to obtain the product (430 g, 60%). LC-MS (ESI): m / z found [MH] - =354.0.

[0930] Synthesis of intermediate 103-3:

[0931] Compound 103-2 (200 mg, 0.563 mmol), iron powder (126 mg, 2.252 mmol), and ammonium chloride (49 mg, 0.912 mmol) were dissolved in 2 mL of H₂O and 8 mL of EtOH at room temperature and allowed to react at 80°C for 5 hours. LC-MS monitoring indicated complete reaction. The reaction solution was filtered through celite, the filtrate was concentrated to dryness, and then separated by normal phase column chromatography (petroleum ether:ethyl acetate = 81:19) to afford the compound (103 mg, 57% yield). MS (ESI): m / z found [MH] - =324.0.

[0932] Synthesis of intermediate 103-5 / 103-6:

[0933] Compound 103-4 (1 g, 3.799 mmol) was dissolved in 10 ml of dry pyridine on an ice bath. TESOTf (8.6 ml, 37.99 mmol) was added and allowed to react in an ice bath for ten minutes before being warmed to room temperature and reacted overnight. LC-MS monitoring showed complete reaction of the starting material. The reaction solution was concentrated under reduced pressure to remove the bulk of pyridine and then dissolved in dichloromethane. The mixture was then extracted three times with water, washed once with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting product was then separated by normal phase column chromatography to afford the compound (1.35 g, 79% yield). MS (ESI): m / z found [M+H] + =378.0 / 492.2.

[0934] Synthesis of intermediate 103-7:

[0935] Compound 103-5 / 103-6 (440 mg, 0.896 mmol) was dissolved in 10 ml of toluene at room temperature. Lawesson's reagent (435 mg, 1.075 mmol) was added. After nitrogen substitution, the temperature was raised to 90°C and the reaction was allowed to proceed for 4 hours. TLC monitoring indicated complete reaction. The reaction solution was concentrated and purified by normal phase column chromatography (petroleum ether:ethyl acetate = 95:5) to afford the compound (153 mg, 43% yield). MS (ESI): m / z found [M+H] + =394.0.

[0936] Synthesis of intermediate 103-8:

[0937] Compound 103-7 (169 mg, 0.430 mmol) was dissolved in 10 ml of dry tetrahydrofuran at room temperature, and triethylamine trihydrofluoride (175 μl, 1.075 mmol) was added. The mixture was allowed to react overnight. TLC monitored the reaction for completion. The reaction solution was diluted with DCM and washed three times with water and once with saturated aqueous NaCl. The mixture was then dried over anhydrous NaSO, concentrated, and purified by column chromatography (petroleum ether:ethyl acetate = 71:29). The product (73 mg, 61% yield) was obtained by distillation under reduced pressure.

[0938] Synthesis of intermediate 103-9:

[0939] Compound 103-3 (34 mg, 0.105 mmol), 103-8 (30 mg, 0.105 mmol), and PPTS (1.3 mg, 0.005 mmol) were dissolved in 10 ml of toluene at room temperature and heated to 110°C for 5 hours. The reaction was complete as monitored by TLC. The reaction solution was concentrated and dissolved in a small amount of dichloromethane and purified by column chromatography (petroleum ether:ethyl acetate = 78:22) to obtain the compound (32 mg, 54% yield). MS (ESI): m / z found [M+H] + =569.0.

[0940] Synthesis of intermediate 103-10:

[0941] Compound 103-9 (68 mg, 0.120 mmol) was dissolved in 5 ml of dry dichloromethane at room temperature, and trifluoroacetic acid (500 μL) was added and allowed to react for 1 hour. TLC monitored the reaction to completion. The reaction solution was concentrated, dissolved in a small amount of DMF, and purified by pre-HPLC (water:acetonitrile = 75:25, 1 / 1000 formic acid) to obtain the compound (12 mg, 22% yield). MS (ESI): m / z found [M+H] + =469.30.

[0942] Synthesis of intermediate 103-11:

[0943] At room temperature, compound Fmoc-L-valine (2.7 mg, 0.0079 mmol), HATU (4.5 mg, 0.0079 mmol), and DIPEA (3.3 μL, 0.0189 mmol) were dissolved in dry DMF. After half an hour of reaction, 103-10 (3.0 mg, 0.0064 mmol) was added and allowed to react overnight at room temperature. LC-MS monitored the reaction for completion. The reaction solution was purified by pre-HPLC (water:acetonitrile = 41.2:58.8) and lyophilized to afford the compound (2.74 mg, 54% yield). MS (ESI): m / z found [M+H] + =790.50.

[0944] Synthesis of intermediate 103-12:

[0945] Compound 103-11 (30 mg, 0.038 mmol) was dissolved in DMF (3 mL) at room temperature, pyridine (1 mL) was added, and the mixture was allowed to react at room temperature for 2 h. After the reaction was complete, the product was purified by pre-HPLC and lyophilized to obtain a yellow solid product (12 mg, 56%). LC-MS (ESI): m / z found [M+H] + =568.32.

[0946] Synthesis of compound 103:

[0947] Compound 57-7 (12 mg, 0.22 mmol) was dissolved in DMF, and HATU (9 mg, 0.023 mmol) and DIEA (8 mg, 0.062 mmol) were added sequentially. The mixture was reacted at room temperature for 2 h. Pre-HPLC purification and lyophilization gave compound 103. LC-MS (ESI): m / z found [M+H] + =1087.4.

[0948] Example 104:

[0949] Synthesis route:

[0950] Synthesis of intermediate 104-1:

[0951] 83-11 (31 mg, 0.106 mmol) was dissolved in DMF (5 mL) at room temperature. HOSU (24 mg, 0.127 mmol) and EDCI (16 mg, 0.127 mmol) were added and reacted for two hours. 97-1 (49 mg, 0.106 mmol) and TEA (16 μL, 0.117 mmol) were then added sequentially and the reaction was maintained at room temperature for one hour. After completion of the reaction, the reaction solution was purified by pre-HPLC and lyophilized to obtain a colorless oil (21 mg, 27%). LC-MS (ESI): m / z found [MH] - =735.40.

[0952] Synthesis of intermediate 104-2:

[0953] Compound 104-1 (21 mg, 0.029 mmol) was dissolved in DMF (3 mL) under ice, and HATU (13 mg, 0.034 mmol) and DIEA (9.9 μL, 0.057 mmol) were added. After reacting for 15 minutes, compound 11 (12 mg, 0.029 mmol) was added. After the reaction was complete, the product was purified by Pre-HPLC and concentrated under reduced pressure to obtain a yellow solid (17 mg, 52%). LC-MS (ESI): m / z found [M+H] + =1144.80.

[0954] Synthesis of compound 104:

[0955] Compounds 83-10 (9.8 mg, 0.012 mmol) and 104-2 (17 mg, 0.015 mmol) were dissolved in THF / EtOH (1:3). CuSO₄.5H₂O (3.1 mg, 0.012 mmol) and ascorbic acid (2.2 mg, 0.012 mmol) were added sequentially. The mixture was reacted at room temperature under nitrogen for 1.5 h. Pre-HPLC preparation and concentration under reduced pressure afforded a yellow solid (6.93 mg, 29.8%). LC-MS (ESI): m / z found [M / 2+H] + =970.20.

[0956] Examples 105-106:

[0957] Except for replacing the corresponding reaction raw materials, Examples 105-106 were synthesized according to the synthesis method of Example 104. Their structures and LC-MS are shown in Table 6.

[0958] Table 6 Structural formula and LC-MS (ESI) of Examples 105-106

[0959] Examples 107-109:

[0960] Synthesis route:

[0961] Synthesis of intermediate 107-2:

[0962] At room temperature, compound 107-1 (200 mg, 0.543 mmol) was dissolved in DCM (5 mL), and 89-2 (303 μL, 5,429 mmol) and PPTS (273 mg, 1.086 mmol) were added sequentially. The mixture was allowed to react overnight at room temperature. After the reaction was complete, the mixture was concentrated under reduced pressure, dissolved in a small amount of DMF, and purified by reverse phase column chromatography. The product was concentrated under reduced pressure to obtain a white solid (200 mg, 99%). LC-MS (ESI): m / z found [M+Na] + =393.0.

[0963] Synthesis of intermediate 107-3:

[0964] 107-2 (20 mg, 0.059 mmol) was dissolved in DMF (2 mL) on an ice bath. (PNP)2CO (6.6 mg, 0.216 mmol) and DIEA (8.3 μL, 0.054 mmol) were added sequentially. The mixture was stirred on ice for 10 min and then allowed to warm to room temperature overnight. Purification by pre-HPLC and concentration under reduced pressure afforded the product as a white solid (9 mg, 31%). 1 H NMR (600MHz, DMSO-d6) δ8.73(t,J=6.7Hz,1H),8.31(d,J=9.1Hz,2H),7.90(d,J=7.6Hz,2H),7.73(dd,J=18.5,8.3Hz,2H),7.63–7.54(m,3H),7.42(t, J=7.4Hz,2H),7.33(t,J=7.4Hz,2H),4.61(d,J=6.7Hz,2H),4.38–4.33(m,2 H), 4.29 (d, J = 7.1Hz, 2H), 4.23 (dd, J = 14.1, 6.8Hz, 1H), 3.70–3.63 (m, 4H).

[0965] Synthesis of intermediate 107-4:

[0966] Compound 107-3 (9 mg, 0.018 mmol) was dissolved in DMF (3 mL), and HOBt (2.8 mg, 0.018 mmol) was added. The mixture was allowed to react at room temperature for 15 min. 11 (8 mg, 0.018 mmol) and DIEA (5.9 μL, 0.034 mmol) were then added sequentially. The reaction was continued at room temperature for two hours. Pre-HPLC preparation and lyophilization yielded the compound. LC-MS (ESI): m / z found [M+H] + =822.50.

[0967] Synthesis of intermediate 107-7:

[0968] Under ice, 83-11 (100 mg, 0.343 mmol) was dissolved in DMF (3 mL). EDCI (79 mg, 0.412 mmol) and HOSU (47 mg, 0.412 mmol) were added sequentially, and the mixture was allowed to react at room temperature for 1 h. 107-6 (96 mg, 0.343 mmol) and TEA (52 μL, 0.377 mmol) were then added, and the mixture was allowed to react at room temperature for 2 h. After the reaction was substantially complete, the mixture was pre-HPLC prepared and lyophilized to obtain a colorless oil (28.1 mg, 15%). LC-MS (ESI): m / z found [M+Na] + =553.50.

[0969] Synthesis of intermediate 107-8:

[0970] Compound 83-10 (34 mg, 0.042 mmol) and compound 107-7 (28.1 mg, 0.051 mmol) were dissolved in THF (2 mL) and EtOH (6 mL). CuSO4·5H2O (11 mg, 0.042 mmol) and ascorbic acid (7.4 mg, 0.042 mmol) were added sequentially. The mixture was reacted at room temperature under N2 protection for 2 h. Pre-HPLC purification and lyophilization afforded a yellow solid. LC-MS (ESI): m / z found [M+H] + =1346.80.

[0971] Except for replacing the corresponding reaction raw materials, the remaining intermediates and Examples 107-109 were synthesized according to the synthesis methods of Example 77 and Example 88. Their structures and LC-MS are shown in Table 7.

[0972] Table 7 Structural formula and LC-MS (ESI) of Examples 105-106

[0973] Example 10: HER2 ADC conjugation

[0974] Interchain disulfide bonds were reduced by adding 1 mM EDTA (Invitrogen, Catalog No. AM9260G) and 8-fold molar equivalents of TECP (Thermo, Catalog No. 77720) to a 10 mg / mL trastuzumab solution, and the mixture was stirred at 37° C., 250 rpm for 2.5 hours.

[0975] Trastuzumab light chain sequence (SEQ ID NO.1):

[0976] Trastuzumab heavy chain sequence (SEQ ID NO.2):

[0977] The mixture was cooled to a target temperature of 4°C and 10 drug equivalents per mole of the compound was added as a 10% (v / v) DMSO solution (Sigma, Catalog No. D2660). After 3 hours, the antibody-drug conjugates were ultrafiltered using PBS. Their numbers and structures are shown in Table 8.

[0978] Table 8 Ligand-drug conjugate numbers and structural formulas

[0979] Note: In Table 8, mAb refers to trastuzumab in this example. Each occurrence of n is an integer or decimal from 1 to 10, representing the average number of units conjugated to each antibody by each selected drug linker compound. n can also be expressed as DAR.

[0980] After ultrafiltration, the concentration of the ADC conjugated product was determined by absorbance at 280 nm and 365 nm. The successful conjugation was confirmed by HPLC-HIC analysis. Antibody purity was determined by SEC-HPLC. DAR values ​​were determined by LC-MS. The results are shown in Table 9.

[0981] Table 9 ADC drug conjugation results of HER2 antibodies

[0982] Example 111: NECTIN-4 ADC Conjugation

[0983] Interchain disulfide bonds were reduced by adding 1 mM EDTA (Invitrogen, Catalog No. AM9260G) and 8-fold molar equivalents of TECP (Thermo, Catalog No. 77720) to a 10 mg / mL PADCEV mAb solution, and the mixture was stirred at 37° C., 250 rpm for 2.5 hours.

[0984] PADCEV monoclonal antibody light chain sequence (SEQ ID NO. 3):

[0985] PADCEV monoclonal antibody heavy chain sequence (SEQ ID NO. 4):

[0986] The mixture was cooled to a target temperature of 4°C and 10 drug equivalents per mole of the compound was added as a 10% (v / v) DMSO solution (Sigma, Cat. No. D2660). After 3 hours, the antibody-drug conjugates (ADCs) were ultrafiltered using PBS. Their names and structures are shown in Table 10.

[0987] Table 10 Names and structural formulas of ligand-drug conjugates

[0988] Note: In Table 10, Ab refers to PADCEV monoclonal antibody in this example. Each occurrence of n is an integer or decimal from 1 to 10, representing the average number of units conjugated to each antibody by each selected drug linker compound. n may also be expressed as DAR.

[0989] After ultrafiltration, the concentration of the ADC conjugated product was determined by absorbance at 280 nm and 365 nm. The successful conjugation was confirmed by HPLC-HIC analysis. Antibody purity was determined by SEC-HPLC. DAR values ​​were determined by LC-MS. The results are shown in Table 11.

[0990] Table 11 ADC drug conjugation results of NECTIN4 antibody

[0991] Example 112: B7H3 ADC Conjugation

[0992] 1. Prepare solution:

[0993] 1) Prepare 1 mM EDTA-PBS buffer by adding 0.5 M EDTA to 20 μL EDTA in 10 ml PBS.

[0994] 2) Dilute 0.5M TCEP to 5mM: 10μL TCEP + 990μL EDTA-PBS.

[0995] 3) Prepare 10% DMSO solution: 100 μL DMSO + 900 μL EDTA-PBS.

[0996] 4) Antibody Ifinatamab was diluted to 10 mg / ml.

[0997] 2. TCEP reduction cleavage of disulfide bonds:

[0998] When synthesizing an ADC with a DAR value of 8: antibody:TCEP = 1:8 (molar ratio);

[0999] 3. Incubate on a 37°C shaker for 2 h (normal shaking speed: 210-230 rpm), then place on ice.

[1000] 4. Dissolve the linker-payload (referring to the linker and small molecule drug portion of the ligand-drug conjugate, excluding the ligand portion): dissolve it in pure DMSO to make a 5 mg / ml solution.

[1001] 5. Add Linker-payload to the antibody at a ratio of antibody: Linker = 1:10 (molar ratio).

[1002] 6. Mix well on a rotary mixer for 2 hours.

[1003] 7. After mixing, desalt using a desalting column.

[1004] 8. For the samples obtained after desalting, the concentration of ADC conjugated product was determined using the BCA method.

[1005] 9. Determine whether the coupling is successful by HPLC-HIC analysis.

[1006] 10. Antibody purity was tested by SEC-HPLC.

[1007] 11. Detect DAR value by LC-MS.

[1008] B7H3 target antibody Ifinatamab heavy chain SEQ ID NO: 5

[1009] B7H3 target antibody Ifinatamab light chain SEQ ID NO: 6

[1010] The names and specific structures of the obtained antibody-drug conjugates are shown in Table 12.

[1011] Table 12 Names and structural formulas of ligand-drug conjugates

[1012] After ultrafiltration, the concentration of the ADC conjugated product was determined by absorbance at 280 nm and 365 nm. The successful conjugation was confirmed by HPLC-HIC analysis. Antibody purity was determined by SEC-HPLC. DAR values ​​were determined by LC-MS. The results are shown in Table 13.

[1013] Table 13 ADC drug conjugation results of B7H3 antibody

[1014] Test Example 1 Test on the inhibition of tumor cell proliferation in vitro by the camptothecin derivatives or analogs (Payload) of the present invention

[1015] The control compounds used in this experiment were DXD (synthesized from EXD according to the synthesis method of patent EP2907824B1. EXD was purchased from MedChemExpress, product number: HY-13631A) and SN38 (purchased from Bidex Pharmaceuticals, product number BD8481). Their structural formulas are

[1016] The cell lines used in this experiment were human breast cancer cells SK-BR-3, T47D, MDA-MB-453, human malignant melanoma cells A375, human lung squamous cell carcinoma cells NCI-H1703, and human gastric cancer cells NCI-N87. Cell suspensions were prepared in fresh cell culture medium containing 10% FBS and then diluted to a density of 5 × 10 4 cells / mL,10×10 4 cells / mL, 100 μL per well was added to a 96-well cell culture plate (Thermo catalog number: 167425), and cultured at 37°C with 5% carbon dioxide for 24 h.

[1017] The test compound of the present invention and the control compound were prepared to 10 mM with DMSO, and the drugs were diluted with 5% FBS 1640 culture medium in a gradient of 50, 10, 2, 0.4, 0.08, 0.016, 0.0032, 0.00064, 0.000128, and 0 μM, with two replicates for each well.

[1018] 50 μL of cell supernatant was pipetted from each 96-well plate, followed by the addition of 50 μL of compound dilutions at final molar masses of 25, 5, 1, 0.2, 0.04, 0.0016, 0.00032, 0.000064, and 0 μM. After mixing, the cells were cultured in a constant temperature CO2 incubator for 4 days. Cell supernatant was then added with 100 μL of detection reagent (5% FBS1640 medium and CCK-8 solution in a 10:1 ratio) and incubated in an incubator for 4 hours. Chemiluminescence was read on a microplate reader (TECAN, Spark), and data were analyzed using Graphpad Prism 8 software.

[1019] The experimental results are shown in Table 14.

[1020] Table 14 IC values ​​of the compounds of the present invention for inhibition of cancer cell proliferation in vitro 50

[1021] Note: “~” indicates IC 50 estimated value.

[1022] Conclusion: The camptothecin derivatives provided by the present invention have better proliferation inhibitory activity against human breast cancer cells SK-BR-3, T47D, MDA-MB-453, human malignant melanoma cells A375, human lung squamous cell carcinoma cells NCI-H1703 and human gastric cancer cells NCI-N87 than DXD and SN38, indicating that they have drug potential.

[1023] Test Example 2 Test on the inhibition of tumor cell proliferation in vitro by the camptothecin derivative (Payload) of the present invention

[1024] 1. Instruments and Reagents

[1025] 2. Experimental Procedure

[1026] a. Plating & culture: MDA-MB-453 (purchased from: Xiehe, catalog number: 1101HUM-PUMCO00016) / T47D (Kebai, CBP60397) / HT1376 cells (purchased from: Kebai, catalog number: CBP60310) were cultured at 1×10 3 / 2×10 3 / 1×10 3 Cells were seeded into 96-well plates at a density of 100 μL, 200 μL of PBS was added to the outermost circle (to reduce the volatilization of the culture medium), and cultured at 37°C, 5% CO2 for 24 h;

[1027] b. Drug Addition & Incubation: Remove the 96-well flat-bottom plate, aspirate the culture medium in the 96-well plate, add 100 μL of ADC drug / payload diluted in culture medium, and incubate at 37°C, 5% CO2 for 5 days.

[1028] 3. Detection and data processing

[1029] a. Remove the 96-well plate, remove the culture medium, and add 100 μL of detection solution (CCK8: culture medium = 1:9) to each well; place in a 37°C, 5% CO2 incubator for 4 hours;

[1030] b. Read the values ​​at 450 nm using a microplate reader (CEY0017) and record them in Excel;

[1031] c. Graphpad Prism 9.0 was used to analyze and organize the data.

[1032] The experimental results are shown in Table 15.

[1033] Table 15 IC values ​​of the camptothecin derivatives of the present invention for inhibition of cancer cell proliferation in vitro 50

[1034] Note: “-” means not tested.

[1035] Conclusion: The camptothecin derivatives provided by the present invention have better proliferation inhibitory activity against human breast cancer cells T47D, MDA-MB-453, and human bladder cancer cells HT1376 than EXD, indicating that they have drug potential.

[1036] Experimental Example 3 In vitro proliferation inhibition test of HER2 target tumor cells by the ligand-drug conjugate of the present invention

[1037] The cell lines used in this experiment were HER2-high-expressing human breast cancer cell line SK-BR-3 and human gastric cancer cell line NCI-N87. Cell suspensions were prepared in fresh cell culture medium containing 10% FBS and then diluted to a density of 5×10 4 cells / mL,10×10 4 cells / mL, 100 μL per well was added to a 96-well cell culture plate (Thermo catalog number: 167425), and cultured at 37°C with 5% carbon dioxide for 24 h.

[1038] The ADC sample was prepared at 10 μM in PBS. This was the initial concentration, and a five-fold gradient dilution was performed using PBS, for a total of nine concentrations. 50 μL of culture medium was aspirated from each well, and 50 μL of the above ADC solution was added, resulting in an initial ADC concentration of 5 μM in each well and a final volume of 100 μL per well. The cells were cultured at 37°C in 5% CO2 for 3 days. 100 μL of CTG ( Luminescent Cell Viability Assay (Promega, Catalog No. G7573) was used. The cells were mixed on a decolorizing shaker at room temperature for 30 minutes, incubated for 10 minutes, and chemiluminescence was read on a microplate reader (TECAN, Spark). Data were analyzed using Graphpad Prism 5 software. The experimental results are shown in Table 16.

[1039] Table 16 IC values ​​of the ligand-drug conjugates of the present invention for inhibition of cancer cell proliferation in vitro 50 value

[1040] Conclusion: The ligand-drug conjugate targeting HER2 of the present invention has significant proliferation inhibitory activity on SK-BR-3 cells and NCI-N87 cells.

[1041] Experimental Example 4 In vitro proliferation inhibition test of NECTIN-4 target tumor cells by the ligand-drug conjugate of the present invention

[1042] The cell lines used in this experiment were human breast cancer T47D and MDA-MB-453 cell lines that highly expressed NECTIN4. Cell suspensions were prepared in fresh cell culture medium containing 10% FBS and then diluted to a density of 5×10 4 cells / mL,10×10 4 cells / mL, 100 μL per well was added to a 96-well cell culture plate (Thermo catalog number: 167425), and cultured at 37°C with 5% carbon dioxide for 24 h.

[1043] The ADC sample was prepared at 10 μM in PBS. This was the initial concentration, and a five-fold gradient dilution was performed using PBS, for a total of nine concentrations. 50 μL of culture medium was aspirated from each well, and 50 μL of the above ADC solution was added, resulting in an initial ADC concentration of 5 μM in each well and a final volume of 100 μL per well. The cells were cultured at 37°C in 5% CO2 for 3 days. 100 μL of CTG ( Luminescent Cell Viability Assay (Promega, Catalog No. G7573) was used. The cells were mixed on a decolorizing shaker at room temperature for 30 minutes, incubated for 10 minutes, and chemiluminescence was read on a microplate reader (TECAN, Spark). Data were analyzed using Graphpad Prism 5 software. The experimental results are shown in Table 17.

[1044] Table 17 IC values ​​of the ligand-drug conjugates of the present invention for inhibition of cancer cell proliferation in vitro 50 value

[1045] Conclusion: The ligand-drug conjugate targeting NECTIN-4 of the present invention has obvious proliferation inhibitory activity on T47D cells and MDA-MB-453 cells.

[1046] Experimental Example 5 In vitro proliferation inhibition test of B7H3 target tumor cells by the ligand-drug conjugate of the present invention

[1047] The cell lines used in this experiment were human A375 (ATCC, Catalog No.: CRL-1619), NCI-H358 (ATCC, Catalog No.: CRL-5807), NCI-H1975 (Kebai, Catalog No.: CBP60121), and NCI-H345 cell lines (BIOBW Beijing Bio-Tech, Catalog No.: bio-133296). Cell suspensions were prepared in fresh cell culture medium containing 10% FBS and then diluted to a density of 2×10 4 cells / mL、4×10 4 cells / mL, 2×10 4 cells / mL and 2×104 cells / mL, 100 μL per well was added to a 96-well cell culture plate (Thermo catalog number: 167425), and cultured at 37°C with 5% carbon dioxide for 24 h.

[1048] The ADC sample was prepared at 10 μM in PBS. This was the initial concentration, and a five-fold gradient dilution was performed using PBS, for a total of nine concentrations. 50 μL of culture medium was aspirated from each well, and 50 μL of the above ADC solution was added, resulting in an initial ADC concentration of 5 μM in each well and a final volume of 100 μL per well. The cells were cultured at 37°C in 5% CO2 for 3 days. 100 μL of CTG ( Luminescent Cell Viability Assay, Promega, catalog number: G7573), mixed on a decolorization shaker at room temperature for 30 minutes, incubated for 10 minutes, and chemiluminescence was read on a microplate reader (TECAN, Spark). Data were analyzed using Graphpad Prism 5 software. The results are shown in Table 18.

[1049] Table 18 IC values ​​of the ligand-drug conjugates of the present invention for inhibition of cancer cell proliferation in vitro 50 value

[1050] Note: “-” means not tested.

[1051] Conclusion: The ligand-drug conjugate provided by the present invention has excellent proliferation inhibitory activity against human A375, NCI-H358, NCI-H1975, and NCI-H345 cell lines with high B7H3 expression, indicating that it has drug potential.

[1052] Test Example 6: Bystander Killing Effect

[1053] 1) In vitro bystander effect:

[1054] 1. Cell culture (Raji(-) or A375(+):Raji(-)=1:1, 1×10 cells of each type 5 ) to a 6-well plate, and 3 ml of 10% FBS1640 medium was added to each well.

[1055] 2. Add the calculated amount of ADC B7H3-ADC-1 and B7H3-ADC-27 to the culture medium and add them to the corresponding wells of the plate so that there is 4 ml of culture medium in each well.

[1056] 3. On the 4th day after drug addition, the supernatant was collected and the cells on the plate were digested. The cells were collected in the same centrifuge tube as the supernatant and centrifuged at 300 × g for 5 min.

[1057] 4. Add FACS to wash cells

[1058] 5. Incubate M30 (1ug / 100ul) antibody in a 4°C refrigerator for 1 hour (antibody + cells = 100ul + 100ul)

[1059] 6. Wash twice with FACS, add PE dye (1:200) and incubate for 30 minutes

[1060] 7. Wash with PBS three times, add NIR dye (1:500) and incubate for 15 minutes

[1061] 8. Wash once with PBS, resuspend in PBS and perform flow cytometry (select PE and APC-cy7 channels)

[1062] Conclusion: The results, shown in Figures 1, 2, 3, and 4, demonstrate that neither the ADC of the present invention nor the positive control ADC exhibited significant killing activity against B7H3-negative cell lines. When co-cultured with B7H3-positive and negative cells, the ADC of the present invention efficiently killed both positive and negative cells, demonstrating a bystander killing effect superior to that of the positive control drug.

[1063] 2) Bystander effect in vivo:

[1064] Experimental methods:

[1065] Prepare B7H3-positive A375 cells and B7H3-negative MDA-MB-453 cells by resuspending them in 100 μl of PBS, adding equal volumes of Matrigel in a 1:1 ratio and mixing thoroughly. Take 200 μl and inoculate subcutaneously into nude mice.

[1066] When grown to 200mm 3 Left and right, tail vein administration, volume 200ul;

[1067] Tumor tissue was removed 15 and 30 days after administration, washed three times with PBS, cut into small pieces, and digested at 37°C for 30 min with 3 ml of trypsin. The tissue was then sieved through a 70 μm cell sieve. Undigested tissue fragments were ground and digested with 6 ml of culture medium containing 10% FBS. The tissue was centrifuged at 200 g for 5 min, resuspended in 1 ml of PBS, and the cells were counted.

[1068] Dilute the cells to 2 × 10 6 / ml, take 50ul, that is 1×10 5 / well cells were plated in a U-bottom 96-well plate;

[1069] Dilute the M30 antibody to 20 μg / ml, take 50 μl (1 μg / well) and add it to the cells, mix well, and incubate at 4°C for 1 hour;

[1070] Wash twice with FACS buffer by centrifugation, add 100 μl of anti-human Fc.PE fluorescent secondary antibody (1:200 dilution), and incubate at 4°C for 30 min;

[1071] Wash twice by centrifugation with FACS washing solution, add 70ul FACS to resuspend, and take 50ul to detect PE fluorescence intensity by flow cytometry.

[1072] Conclusion: The results are shown in Figure 5, indicating that the ADC of the present invention exhibits superior tumor inhibition activity compared to positive drug ADCs in an animal model in which both B7H3-negative and positive cells coexist. The tumor (volume) inhibition rate (TGI) of the control ADC B7H3-ADC-12mpk was 58.37%, and that of B7H3-ADC-22mpk was 54.81%. The TGI of the ADC provided by the present invention, B7H3-ADC-12mpk, was 90.46%, and that of 1mpk was 71.95%, indicating that the ADC of the present invention has a bystander effect that is significantly superior to that of positive drugs.

[1073] Experimental Example 7 Antitumor Efficacy of the B7H3 Ligand-Drug Conjugate of the Present Invention in the A375 CDX Model

[1074] Experimental steps:

[1075] 1. Human malignant melanoma cells A375 (ATCC) (5×10 6 After cell inoculation, the tumors grew for 9 days and the tumor volume reached 200 mm. 3 After the experiment, the animals were randomly divided into groups (D0), with 5 animals in each group, for a total of 5 groups.

[1076] 2. B7H3-ADC-1 and B7H3-ADC-27 were administered via tail vein injection at two doses, 2 mg / kg and 4 mg / kg, respectively, as a single dose. Tumor volume and body weight were measured twice weekly and the data were recorded. Tumor volume (V): V = 1 / 2 × major diameter × minor diameter 2 The tumor growth curve was prepared using GrapHPadPrism 8.0.2.2.263 software. The tumor growth curve is shown in FIG6 , and the changes in mouse weight are shown in FIG7 .

[1077] Conclusion: The ADC B7H3-ADC-27 provided by the present invention can significantly inhibit the growth of tumors in A375 model mice at both 2mpk and 4mpk doses, and its anti-tumor activity is significantly better than that of the positive control ADC. In addition, no animal deaths or significant weight loss were observed in all treatment groups during the observation period, indicating that B7H3-ADC-27 has no obvious toxicity.

[1078] Experimental Example 8 Antitumor efficacy of the NECTIN4 ligand-drug conjugate of the present invention in the MDA-MB-453CDX model

[1079] 1. Human breast cancer cells MDA-MB-453 (Xiehe) (1×10 7 After cell inoculation, the tumor grew for 9 days and the tumor volume reached 150 mm. 3 After the experiment, the animals were randomly divided into three groups (D0), with 5 animals in each group, for a total of 3 groups.

[1080] 2. NECTIN4-ADC-1 and NECTIN4-ADC-2 were administered via tail vein injection at a single dose of 5 mg / kg. Tumor volume and body weight were measured twice weekly and recorded. Data were analyzed using Excel 2016 statistical software: mean was calculated as average; SD was calculated as STDEV; and SEM was calculated as STDEV / SQRT. Tumor growth curves were generated using GraphPad Prism 8.0.2.2.263 software.

[1081] Tumor volume (V): V = 1 / 2 × L long diameter × L short diameter 2

[1082] Relative tumor volume (RTV): RTV = VT / V0

[1083] Relative tumor proliferation rate T / C (%) = TRTV / CRTV × 100%

[1084] Tumor inhibition rate (%) = (CRTV-TRTV) / CRTV (%)

[1085] Where V0 and VT are the tumor volumes at the start and end of the experiment, respectively. CRTV and TRTV are the relative tumor volumes of the blank control group (PBS) and the experimental group at the end of the experiment, respectively. The changes in tumor volume of the mice are shown in Figure 8, and the changes in mouse body weight are shown in Figure 9.

[1086] Conclusion: The body weight of mice increased steadily, with no significant safety risks. Furthermore, NECTIN4-ADC-2 (5 mg / kg) was significantly more effective than the control drug NECTIN4-ADC-1 (5 mg / kg).

[1087] Test Example 9 Toxicity Test of Ligand-Drug Conjugates of the Present Invention

[1088] 1. Experimental purpose:

[1089] The potential toxicity of ADC administered five times via tail vein injection as a single dose in Balb / c mice was studied, and the dosage and frequency of administration were preliminarily determined to provide a reference for subsequent formal toxicology experiments.

[1090] 2. Experimental Animals

[1091] Species / strain / grade: Balb / c mice, SPF grade

[1092] Experimental animals: Source: Weitonglihua. Age: 6-8 weeks. Number and gender: 8 females.

[1093] 3. Experimental Design

[1094] 3.1 Trial Grouping

[1095] 3.2 Experimental methods

[1096] Administration route and frequency: intravenous injection, once a week, for a total of 5 doses, Day 0, 7, 14, 21, 28

[1097] Experimental duration: 6 weeks

[1098] Dosing volume: 170μL-210μL

[1099] 3.3 Detection indicators:

[1100] ① General clinical observation: once before grouping and once a day during the trial.

[1101] ② Body weight: Weigh once before administration and twice weekly thereafter. Weigh once before planned euthanasia of animals in the main experimental group.

[1102] ③ Histopathological examination: Based on the gross observation results and possible toxic target organs, the heart, liver, spleen, lung, kidney, and small intestine were prepared and examined. The specific experimental procedures are as follows:

[1103] a. Sample: Tissue sample, approximately 3mm thick.

[1104] b. Dehydration: Dehydration with graded alcohol: 60%, 70%, 80%, 90%, 95% I, 95% II, 100% I, 100% II for 1 hour per tank; xylene I for 30 minutes, xylene II for 20 minutes; paraffin wax immersion: paraffin I and paraffin II for 20 minutes per tank.

[1105] c. Embedding: Paraffin embedding.

[1106] d. Section: The thickness of the section is about 4 μm.

[1107] e. Hematoxylin eosin (HE) staining: ① Dewaxing and hydrating: dewaxing in xylene I and xylene II for 15 min each; 100% anhydrous ethanol, 95% ethanol, 80% ethanol, and 70% ethanol for 10 min each; rinsing with running water for 2 min; ② Hematoxylin staining for 10 min, followed by rinsing with running water; ③ 0.5% eosin staining for 5 min, followed by rinsing with running water; ④ Gradient alcohol dehydration: 70%, 80%, 95%, and 100%, 10 s each; ⑤ Xylene clearing: xylene I and xylene II for 5 min each; ⑥ Seal with neutral resin glue.

[1108] f. Observe under light microscope and take microphotographs.

[1109] 3.4 Data Analysis

[1110] The experimental results are expressed as mean ± standard error, and one-way analysis of variance was performed. P values ​​less than 0.05 were considered statistically significant (*p<0.05).

[1111] 4. Experimental results:

[1112] HE, -: no obvious lesions, +: mild lesions, ++: mild, +++: moderate, ++++: severe.

[1113] The changes in mouse body weight are shown in FIG10 .

[1114] Conclusion: For the test substance B7H3-ADC-27, compared with the vehicle group, at a dose of 70 mg / kg, the animal body weight did not decrease significantly, and showed a slow growth state (p < 0.05), indicating that the ligand drug conjugate provided by the present invention has no obvious toxicity.

[1115] Experimental Example 10 Antitumor efficacy of the B7H3 ligand-drug conjugate of the present invention in the NCI-H358 CDX model

[1116] 1. Experimental Procedure

[1117] BALB / c-Nude mice were subcutaneously inoculated with human non-small cell lung cancer cells NCI-H358 (3×106 / 200 μL / mouse, in 50% low-growth factor artificial basement membrane matrix gel) in the right flank. After cell inoculation, the tumor grew for 5 days and the tumor volume reached 130 mm. 3 After about 24 hours, the animals were randomly divided into 13 groups (D0), with 5 animals per group. Drugs were administered via tail vein injection, with high and low doses given once. Tumor volume and body weight were measured twice weekly and the data were recorded.

[1118] 2. Data Analysis

[1119] Data were analyzed using Excel 2016 statistical software, with averages calculated as "average." Tumor growth curves were generated using GraphHPad Prism 8.0.2.2.263 software.

[1120] Tumor volume (V): V = 1 / 2 × L long diameter × L short diameter 2

[1121] Relative tumor volume (RTV) = VT / V0

[1122] Relative tumor proliferation rate T / C (%) = TRTV / CRTV × 100%

[1123] Tumor inhibition rate (%) = (CRTV-TRTV) / CRTV (%)

[1124] Where V0 and VT are the tumor volumes at the start and end of the experiment, respectively. CRTV and TRTV are the relative tumor volumes of the blank control (PBS) and experimental groups at the end of the experiment, respectively. The changes in tumor volume in the mice are shown in Figures 11 and 12.

[1125] Conclusion: The experimental group ADCs B7H3-ADC-15, B7H3-ADC-21, B7H3-ADC-23 (DAR4), and B7H3-ADC-27 provided by the present invention can significantly inhibit the growth of tumors in NCI-H358 model mice, whether in the low-dose group or the high-dose group, and their anti-tumor activity is significantly better than that of the positive control ADCs (B7H3-ADC-1, B7H3-ADC-2-DAR4).

[1126] The present invention has been described through the above-described embodiments. However, it should be understood that the above-described embodiments are for illustrative and illustrative purposes only and are not intended to limit the present invention to the described embodiments. Furthermore, it will be understood by those skilled in the art that the present invention is not limited to the above-described embodiments and that further variations and modifications may be made based on the teachings of the present invention, all of which fall within the scope of the present invention. The scope of protection of the present invention is defined by the appended claims and their equivalents.

Claims

1. A compound as shown in Formula 1 or a pharmaceutically acceptable salt thereof, in, R 16 , R 17 , R 19 Each occurrence is independently selected from hydrogen, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 Alkynyl; preferably, R 16 , R 17 , R 19 Each occurrence is independently selected from hydrogen and C optionally substituted by R 1-6 More preferably, R 16 , R 17 , R 19 Each occurrence is independently selected from hydrogen and C 1-6 Alkyl; further preferably, R 16 , R 17 , R 19 Each occurrence of is independently hydrogen; R 20 Selected from N, and CR 20a ; R 20a selected from hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -C(O)CH2OH, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, and C 1-6 Alkyl; preferably, R 20a is hydrogen or C 1-6 alkyl; R 11 Selected from O and S; Q is selected from 0, 1, 2, 3, and 4; R 18 is selected from H, and -B-R3; When Q is 0, R 18 When it is -B-R3, R 14 and R 15 are independently selected from hydrogen atoms, halogens, C 1-8 Alkyl, and C 1-8 Preferably, R 14 and R 15 Each independently selected from hydrogen, F, Cl, Br, I, C 1-3 Alkyl and C 1-3 Preferably, R 14 and R 15 Each independently is -CH3 or F; preferably, R 14 is -CH3, and / or R 15 for -F; B is selected from -C(O)- and -P(O)(OH)-; preferably B is -C(O)-; R3 is selected from -(CH2CH2O) a C 1-6 alkyl, When -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m -n methylene units are each independently replaced by -N(R5)C(O)-, -C(O)-, -OC(O)-, -NR5-, -O-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -C(=S)-, -C(=NR5)-, -N=N- or -N=CH-; a is an integer greater than or equal to 1, preferably, a is an integer from 1 to 20, and more preferably, a is selected from 1, 2, 3, 4, 5, 6, 7, and 8; R4 and R8 are each independently a single bond or are independently selected from -O-, C 1-8 Alkylene, C 1-8 Haloalkylene, -N(R5)C(O)-, -C(O)-, -OC(O)-, -NR5-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -C(=S)-, -C(=NR5)-, -N=N- and -N=CR5-; Ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from 6-10 membered aryl or arylene, 5-10 membered heteroaryl or heteroarylene, 3-10 membered cycloalkyl or cycloalkylene, and 3-10 membered heterocyclyl or heterocyclylene, preferably, the heteroaryl, heteroarylene, heterocyclylene and heterocyclyl each independently contain 1, 2, 3 or 4 heteroatoms independently selected from N, O, P and S; and when R3 is When W1 and W2 are fused together, when R3 is When W1 and W2 are fused together, W2 and W3 are fused together, and W3 and W4 are fused together; When ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from 3-10 membered cycloalkyl, cycloalkylene, heterocyclylene, and heterocyclyl, the 3-10 membered cycloalkyl, cycloalkylene, heterocyclylene, and heterocyclyl are optionally replaced by one or more R w1 , R w2 , R w3 , R w4 , or R w5 Replacement, R w1 , R w2 , R w3 , R w4 , and R w5 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halogen, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Alkyl; preferably, R w1 , R w2 , R w3 , R w4 , and R w5 Each time it appears independently selected from hydrogen, deuterium, F, Cl, Br, I, hydroxyl, =O, =S, -NH2, -NO2, -CN, -COOH, -OC 1-3 Alkyl, -C 1-3 Alkyl, and C 1-3 Alkylene hydroxyl group; When ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from 6-10 membered aryl, 6-10 membered arylene, 5-10 membered heteroarylene, and 5-10 membered heteroaryl, the aryl, arylene, heteroarylene, and heteroaryl groups are optionally replaced by one or more R w1 , R w2 , R w3 , R w4 , or R w5 Replacement, R w1 , R w2 , R w3 , R w4 , and R w5 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Alkyl; preferably, R w1 , R w2 , R w3 , R w4 , and R w5 Each occurrence is independently selected from hydrogen, deuterium, F, Cl, Br, I, hydroxyl, -NH2, -NO2, -CN, -OC 1-3 Alkyl, -COOH, and C 1-3 alkyl; R 13 Selected from hydrogen atoms, halogens, C 1-8 Alkyl, and C 1-8 Preferably, R 13 is a hydrogen atom; R 12 Selected from C 1-8 Alkylene N(R a )(R b ), C 1-8 Haloalkylene N(R a )(R b ), halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Alkyl; preferably, R 12 Selected from C 1-3 Alkylene N(R a )(R b ), and C 1-3 Haloalkylene N(R a )(R b ); preferably, R 12 Selected from -CH2N(R a )(R b ); or, R 13 and R 12 Together with the carbon atom to which it is attached, it forms a saturated or unsaturated 5-10 membered cycloalkyl or a 5-10 membered heterocyclic group containing 1, 2 or 3 heteroatoms independently selected from N, O, P and S, wherein the cycloalkyl or heterocyclic group is optionally substituted by one or more independently selected from halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 The alkyl group is substituted with a substituent; preferably, R 13 and R 12 Together with the carbon atom to which it is attached, a 6-membered cycloalkyl is formed, wherein the cycloalkyl is -N(R a )(R b )replace; R 3a , R 3b , and R5, at each occurrence, is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Alkyl; and m, and n are each independently an integer greater than or equal to 1, and m≥n; preferably, m and n are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m≥n; preferably, m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, -n is 0, 1, 2, 3, 4, or 5, and m≥n; When Q is 1, 2, 3 or 4, R 18 When H, R 13 , R 14 , and R 15 are each independently selected from hydrogen, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Alkyl; preferably, R 13 , R 14 , and R 154 Each independently selected from hydrogen, F, Cl, Br, I, C 1-3 Alkyl, -NO2, -CN, -OC 1-3 Alkyl; or R 14 and R 15 Together with the carbon atom to which it is attached, it forms a saturated or unsaturated 5-10 membered cycloalkyl or a 5-10 membered heterocyclic group containing 1, 2 or 3 heteroatoms independently selected from N, O, and S, wherein the cycloalkyl or heterocyclic group is unsubstituted or substituted by one or more independently selected from halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 The alkyl group is substituted with a substituent; preferably, R 14 and R 15 With The carbon atoms connected together form a saturated or unsaturated 5-6-membered heterocyclic group containing 2 O atoms; R 12 C 1-8 Alkylene hydroxyl, C 1-8 Alkyleneamino, -C(R a )(R b )-N(R a )-R 4c ,-C(R a )(R b )-N(R a )-C(O)-R 4c , or -C(R a )(R b )-N(R a )-C(O)OR 4c Preferably, R 12 -CH2-NH2, -CH2-NH-C(O)-R 4c 、-CH2-NH-C(O)OR 4c , or C4H8OH; preferably, R 12 is -CH2-NH2, or -(CH2)4-OH; and R, R a , R b , and R 4c Each occurrence is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -C(O)CH2OH, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 Alkyl or C 1-6 Alkylene hydroxyl group; preferably, R, R a , R b ,R 4c Each occurrence is independently hydrogen, F, Cl, Br, I, CH2OH, or C2H4OH; When Q is 0, R 11 is S, and R 18 When H, R 12 is selected from hydrogen, and -CH2N(R 3A )(R 4A ); R 14 , and R 15 are each independently selected from hydrogen, halogen, -NO2, -CN, -OR', -N(R a1 )(R b1 ), -C(O)R', -CO2R', -C(O)C(O)R', -C(O)CH2C(O)R', -C(O)N(R a1 )(R b1 )、-SO2N(R a1 )(R b1 ), -OC(O)R', -N(R')SO2R', and C 1-6 Alkyl; preferably, R 14 and R 15 are independently selected from halogen, amino, -OR' and C 1-6 Alkyl; further preferably, R 14 and R 15 Each independently selected from fluorine, chlorine, amino, -OC 1-6 Alkyl and C 1-6 Alkyl; or R 14 and R 15 can together form a saturated or unsaturated 4-10-membered cycloalkyl or a 5-membered or 6-membered heterocyclic group containing 1-3 heteroatoms independently selected from N, O, and S, wherein the cycloalkyl or heterocyclic group is unsubstituted or substituted by one or more independently selected from halogen, -NO2, -CN, -OR, -SR, -N(R a1 )(R b1 ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a1 )(R b1 )、-SO2N(R a1 )(R b1 ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl substitution; preferably, the cycloalkyl or heterocyclyl is unsubstituted; R 13 For hydrogen, -C 1-6 Alkyl or -C 1-6 Alkylene-hydroxy; preferably, R 5A For hydrogen, C 1-6 Alkyl or -(CH2)4-hydroxy; R 3A and R 4A Each occurrence is independently selected from hydrogen, halogen, -OR', -N(R a1 )(R b1 ), -C(O)R', -C(S)R', -C(S)C(R a1 )(R b1 )OH、-C(O)CH2N(R a1 )(R b1 ), -CO2R', -C(O)C(O)R', -C(O)CH2C(O)R', -C(O)C(R a1 )(R b1 )OH, -C(O)-C 3-6 Cycloalkylene-OH, -C(O)-C 1-6 Alkylene -OH, -C 1-6 Alkylene-OH-C(O)-C 3-6 Cycloalkylene-C 1-6 Alkylene -OH, -C(O)OC 1-6 Alkylene -OH, -S(O)R', -S(O)2R, -C(O)N(R a1 )(R b1 )、-SO2N(R a1 )(R b1 ), -OC(O)R', -N(R')SO2R', and C optionally substituted by R' 1-6 Alkyl, C 1-6 Alkenyl and C 1-6 Alkynyl; preferably, R 4A Selected from hydrogen, or C 1-6 alkyl; Where R', R a1 , and R b1 Each occurrence is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, 3, 4, 5, 6, 7 or 8 membered cycloalkyl, 3-8 membered heterocyclyl containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, 6-10 membered aryl, or 5-10 membered heteroaryl containing 1, 2, or 3 heteroatoms independently selected from N, O, and S; the C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl, 3, 4, 5, 6, 7 or 8 membered cycloalkyl, 3-8 membered heterocyclyl, 6-10 membered aryl or 5-10 membered heteroaryl are unsubstituted or substituted by one or more independently selected from halogen, -NO2, -CN, -OH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 Alkyl, C 2-6 Alkenyl or C 1-6 Alkynyl, and hydroxyl C 1-6 Alkylene substitution; When R 14 -CH3, R 15 is fluorine, and R 13 When it is hydrogen, R 12 for When R 14 for -O-CH3, R 15 is fluorine, R 13 When it is hydrogen, R 12 Not for and When R 14 With R 15 Forming 1,4-dioxo six-membered ring, R 13 When it is hydrogen, R 12 -CH2NH2, -CH2NHC2H4OH, -CH2N(CH3)C2H4OH, -CH2NHC3H6OH, and When R 14 is fluorine, R 15 is fluorine, R 13 When it is hydrogen, R 12 is not hydrogen; and When R 14 -CH3, R 15 is chlorine, R 13 When it is hydrogen, R 12 Selected from and When R 14 With R 15 Forming a 1,3-dioxo five-membered ring, R 13 When it is hydrogen, R 12 Selected from -CH2NHC2H4OH, -CH2N(CH3)C2H4OH, -CH2NHC3H6OH, or and When R 13 When selected from -(CH2)4-OH, R 12 When it is hydrogen, R 14 , R 15 is fluorine, or R 14 With R 15 It forms a 5- or 6-membered heterocyclic ring containing two oxygen atoms.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the structure of Formula 1 is selected from the following structures: When Formula 1 is Formula 2A: in, R1 and R2 are each independently selected from hydrogen atoms, halogens, C 1-8 Alkyl, and C 1-8 Preferably, R1 and R2 are each independently selected from hydrogen atom, F, Cl, Br, I, C 1-3 Alkyl and C 1-3 Haloalkyl; preferably, R1 and R2 are each independently -CH3 or F; preferably, R1 is -CH3, and / or R2 is F; A is selected from oxygen atoms and sulfur atoms; B is selected from -C(O)- and -P(O)(OH)-; preferably B is -C(O)-; R3 is selected from -(CH2CH2O) a C 1-6 alkyl, When -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m -n methylene units are each independently replaced by -N(R5)C(O)-, -C(O)-, -OC(O)-, -NR5-, -O-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -C(=S)-, -C(=NR5)-, -N=N- or -N=CH-; a is an integer greater than or equal to 1, preferably, a is an integer from 1 to 20, and more preferably, a is selected from 1, 2, 3, 4, 5, 6, 7, and 8; R4 and R8 are each independently a single bond or are independently selected from -O-, C 1-8 Alkylene, C 1-8 Haloalkylene, -N(R5)C(O)-, -C(O)-, -OC(O)-, -NR5-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -C(=S)-, -C(=NR5)-, -N=N- and -N=CR5-; Ring W1, Ring W2, Ring W3, Ring W4, and Ring W5 are each independently selected from 6-10 5-10 membered aryl or arylene, 5-10 membered heteroaryl or heteroarylene, 3-10 membered cycloalkyl or cycloalkylene, and 3-10 membered heterocyclyl or heterocyclylene, preferably, the heteroaryl, heteroarylene, heterocyclylene and heterocyclyl each independently contain 1, 2, 3 or 4 heteroatoms independently selected from N, O, P and S; and when R3 is When W1 and W2 are fused together, when R3 is When W1 and W2 are fused together, W2 and W3 are fused together, and W3 and W4 are fused together; When ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from 3-10 membered cycloalkyl, cycloalkylene, heterocyclylene, and heterocyclyl, the 3-10 membered cycloalkyl, cycloalkylene, heterocyclylene, and heterocyclyl are optionally replaced by one or more R w1 , R w2 , R w3 , R w4 , or R w5 Replacement, R w1 , R w2 , R w3 , R w4 , and R w5 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halogen, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Alkyl; preferably, R w1 , R w2 , R w3 , R w4 , and R w5 Each occurrence is independently selected from hydrogen, deuterium, F, Cl, Br, I, hydroxyl, =O, =S, -NH2, -NO2, -CN, -COOH, -OC 1-3 Alkyl, -C 1-3 Alkyl, and C 1-3 Alkylene hydroxyl group; When ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from 6-10 membered aryl, 6-10 membered arylene, 5-10 membered heteroarylene, and 5-10 membered heteroaryl, the aryl, arylene, heteroarylene, and heteroaryl groups are optionally replaced by one or more R w1 , R w2 , R w3 , R w4 , or R w5 Replacement, R w1 , R w2 , R w3 , R w4 , and R w5 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Alkyl; preferably, R w1 , R w2 , R w3 , R w4 , and R w5 Each occurrence is independently selected from hydrogen, deuterium, F, Cl, Br, I, hydroxyl, -NH2, -NO2, -CN, -OC 1-3 Alkyl, -COOH, and C 1-3 alkyl; R6 is selected from hydrogen atom, halogen, C 1-8 Alkyl, and C 1-8 Preferably, R6 is a hydrogen atom; R7 is selected from C 1-8 Alkylene N(R a )(R b ), C 1-8 Haloalkylene N(R a )(R b ), halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, -O-CH2C(O)N(R a )(R b ) and C optionally substituted by R 1-6 Alkyl; preferably, R7 is selected from C 1-3 Alkylene N(R a )(R b ), and C 1-3 Haloalkylene N(R a )(R b ); Preferably, R7 is selected from -CH2N(R a )(R b ); or, R6 and R7 together with the carbon atom to which they are attached form a saturated or unsaturated 5-10 membered cycloalkyl or a 5-10 membered heterocyclic group containing 1, 2 or 3 heteroatoms independently selected from N, O, P and S, wherein the cycloalkyl or heterocyclic group is optionally substituted by one or more independently selected from halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Preferably, R6 and R7 form a 6-membered cycloalkyl group with the carbon atom to which they are attached, and the cycloalkyl group is -N(R a )(R b )replace; R 3a , R 3b , and R5, at each occurrence, is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 alkyl; R, R a , and R b Each occurrence is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -C(O)CH2OH, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 Alkylene hydroxyl or C 1-6 Alkyl; and m, and n are each independently an integer greater than or equal to 1, and m≥n; preferably, m and n are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m≥n; preferably, m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, -n is 0, 1, 2, 3, 4, or 5, and m≥n; When Formula 1 is Formula 2B: Where q is 1, 2 or 3; R 1c , R 2c , and R 5c are each independently selected from hydrogen, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 Alkyl; preferably, R 1c , R 2c , and R 5c Each independently selected from hydrogen, F, Cl, Br, I, C 1-3 Alkyl, -NO2, -CN, -OC 1-3 Alkyl; or R 1c and R 2c Together with the carbon atom to which it is attached, it forms a saturated or unsaturated 5-10 membered cycloalkyl or a 5-10 membered heterocyclic group containing 1, 2 or 3 heteroatoms independently selected from N, O, and S, wherein the cycloalkyl or heterocyclic group is unsubstituted or substituted by one or more independently selected from halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, and C optionally substituted by R 1-6 The alkyl group is substituted with a substituent; preferably, R 1c and R 2c Together with the carbon atom to which it is attached, it forms a saturated or unsaturated 5-6-membered heterocyclic group containing 2 O atoms; R 6c C 1-8 Alkylene hydroxyl, C 1-8 Alkyleneamino, -C(R a )(R b )-N(R a )-R 4c ,-C(R a )(R b )-N(R a )-C(O)-R 4c , or -C(R a )(R b )-N(R a )-C(O)OR 4c Preferably, R 6c -CH2-NH2, -CH2-NH-C(O)-R 4c 、-CH2-NH-C(O)OR 4c , or C4H8OH; preferably, R 6c is -CH2-NH2, or -(CH2)4-OH; and R, R a , R b , and R 4c Each occurrence is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -C(O)CH2OH, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 Alkyl or C 1-6 Alkylene hydroxyl group; preferably, R, R a , R b ,R 4c Each occurrence is independently hydrogen, F, Cl, Br, I, CH2OH, or C2H4OH; When Formula 1 is Formula 2C: R 6A is selected from hydrogen, and -CH2N(R 3A )(R 4A ); R 1A , and R 2A are each independently selected from hydrogen, halogen, -NO2, -CN, -OR', -N(R a1 )(R b1 ), -C(O)R', -CO2R', -C(O)C(O)R', -C(O)CH2C(O)R', -C(O)N(R a1 )(R b1 )、-SO2N(R a1 )(R b1 ), -OC(O)R', -N(R')SO2R', and C 1-6 Alkyl; preferably, R 1A and R 2A are independently selected from halogen, amino, -OR' and C 1-6 Alkyl; further preferably, R 1A and R 2A Each independently selected from fluorine, chlorine, amino, -OC 1-6 Alkyl and C 1-6 alkyl; or R 1A and R 2A can together form a saturated or unsaturated 4-10-membered cycloalkyl or a 5-membered or 6-membered heterocyclic group containing 1-3 heteroatoms independently selected from N, O, and S, wherein the cycloalkyl or heterocyclic group is unsubstituted or substituted by one or more independently selected from halogen, -NO2, -CN, -OR, -SR, -N(R a1 )(R b1 ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a1 )(R b1 )、-SO2N(R a1 )(R b1 ), -OC(O)R, -N(R)SO2R, or C1-6 alkyl, C1-6 alkenyl or C1-6 alkynyl optionally substituted by R; preferably, the cycloalkyl or heterocyclic group is unsubstituted; R 5A is hydrogen or C 1-6 Alkyl or -C 1-6 Alkylene-hydroxy; preferably, R 5A For hydrogen, C 1-6 Alkyl or -(CH2)4-hydroxy; R 3A and R 4A Each occurrence is independently selected from hydrogen, halogen, -OR', -N(R a1 )(R b1 ), -C(O)R', -C(S)R', -C(S)C(R a1 )(R b1 )OH、-C(O)CH2N(R a1 )(R b1 ), -CO2R', -C(O)C(O)R', -C(O)CH2C(O)R', -C(O)C(R a1 )(R b1 )OH, -C(O)-C 3-6 Cycloalkylene-OH, -C(O)-C 1-6 Alkylene -OH, -C(O)-C 3-6 Cycloalkylene-C 1-6 Alkylene -OH, -C(O)OC 1-6 Alkylene -OH, -S(O)R', -S(O)2R, -C(O)N(R a1 )(R b1 )、-SO2N(R a1 )(R b1 ), -OC(O)R', -N(R')SO2R', and C optionally substituted by R' 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 Alkynyl; preferably, R 4A Selected from hydrogen, or C 1-6 alkyl; Where R', R a1 , and R b1 Each occurrence is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, 3, 4, 5, 6, 7 or 8 membered cycloalkyl, 3-8 membered heterocyclyl containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, 6-10 membered aryl, or 5-10 membered heteroaryl containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, wherein the C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 The alkynyl, 3-, 4-, 5-, 6-, 7-, or 8-membered cycloalkyl, 3- to 8-membered heterocyclyl, 6- to 10-membered aryl, or 5- to 10-membered heteroaryl is unsubstituted or substituted with one or more substituted alkyl radicals independently selected from halogen, -NO2, -CN, -OH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C ... 1-6 Alkyl, C 2-6 Alkenyl or C 1-6 Alkynyl, and hydroxyl C 1-6 Alkylene substitution; When R 1A -CH3, R 2A is fluorine, R 5A When it is hydrogen, R 6A for and When R 1A for -O-CH3, R 2A is fluorine, R 5A When it is hydrogen, R 6A Not for and When R 1A With R 2A Forming 1,4-dioxo six-membered ring, R 5A When it is hydrogen, R 6A -CH2NH2, -CH2NHC2H4OH, -CH2N(CH3)C2H4OH, -CH2NHC3H6OH, and When R 1A is fluorine, R 2A is fluorine, R 5A When it is hydrogen, R 6A is not hydrogen; and When R 1A -CH3, R 2A is chlorine, R 5A When it is hydrogen, R 6A Selected from and When R 1A With R 2A Forming a 1,3-dioxo five-membered ring, R 5A When it is hydrogen, R 6A Selected from -CH2NHC2H4OH, -CH2N(CH3)C2H4OH, -CH2NHC3H6OH, or and Among them, when R 5A When selected from -(CH2)4-hydroxy, R 6A is hydrogen, R 1A , R 2A is fluorine, or R 1A With R 2A It forms a 5- or 6-membered heterocyclic ring containing two oxygen atoms.

3. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein: When Formula 1 is Formula 2A: R1 and R2 are each independently selected from hydrogen atoms, F, Cl, Br, I, C 1-3 Alkyl and C 1-3 Preferably, R1 and R2 are each independently -CH3 or F; Preferably, R1 is -CH3, and / or R2 is F; and / or a is an integer from 1 to 20, more preferably, a is selected from 1, 2, 3, 4, 5, 6, 7, and 8; and / or R6 is a hydrogen atom, and R7 is selected from C 1-3 Alkylene N(R a )(R b ), and C 1-3 Haloalkylene N(R a )(R b ); Preferably, R7 is selected from -CH2N(R a )(R b ); or R6 and R7 form a 6-membered cycloalkyl with the carbon atom to which they are connected, wherein the cycloalkyl is -N(R a )(R b ); and / or m and n are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m ≥ n; preferably, m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, -n is 0, 1, 2, 3, 4, or 5, and m ≥ n; and / or Ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from 6- or 8-membered aryl or arylene, 5-, 6-, 7- or 8-membered heteroaryl or heteroarylene containing 1, 2, 3 or 4 heteroatoms independently selected from N, O, and S, 4-, 5-, 6-, 7- or 8-membered cycloalkyl or cycloalkylene, and 4-, 5-, 6-, 7- or 8-membered heterocyclyl or heterocyclylene containing 1, 2, 3 or 4 heteroatoms independently selected from N, O, and S. When ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from 4-8 membered cycloalkyl, cycloalkylene, heterocyclylene, and heterocyclyl, the 4-, 5-, 6-, 7- or 8-membered cycloalkyl, cycloalkylene, heterocyclylene, and heterocyclyl are optionally substituted with one or more R w1 , R w2 , R w3 , R w4 , or R w5 Replacement, R w1 , R w2 , R w3 , R w4 , and R w5 Each occurrence is independently selected from hydrogen, deuterium, F, Cl, Br, I, hydroxyl, =O, =S, -NH2, -NO2, -CN, -COOH, -OC 1-3 Alkyl, -C 1-3 Alkyl, and C 1-3 Alkylene hydroxyl group; when ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from 6 or 8-membered aryl, 6 or 8-membered arylene, 5, 6, 7 or 8-membered heteroarylene and heteroaryl, the aryl, arylene, heteroarylene and heteroaryl are optionally substituted by one or more R w1 , R w2 , R w3 , R w4 , or R w5 Replacement, R w1 , R w2 , R w3 , R w4 , and R w5 Each occurrence is independently selected from hydrogen, deuterium, F, Cl, Br, I, hydroxyl, -NH2, -NO2, -CN, -OC 1-3 Alkyl, -COOH, and C 1-3 Preferably, ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently optionally substituted with hydrogen, deuterium, F, Cl, Br, I, hydroxyl, -NH2, -NO2, -CN, -OC 1-3 Alkyl, -COOH, or C 1-3 Alkyl-substituted phenyl or phenylene, pyridyl or pyridylene, pyrazolyl or pyrazolylene, or or optionally substituted with hydrogen, deuterium, F, Cl, Br, I, hydroxyl, =O, =S, -NH2, -NO2, -CN, -COOH, -OC 1-3 Alkyl, -C 1-3 Alkyl, or C 1-3 Alkylene hydroxy-substituted tetrahydropyranyl or tetrahydropyranylene, dioxolanyl or dioxolanylene, tetrahydrofuranyl or tetrahydrofuranylene, indicates a site of attachment; and / or When R3 is When W1 and W2 are fused together to form the following fused ring structure: Among them, R w1 and R w2 Each occurrence is independently selected from hydrogen, deuterium, F, Cl, Br, I, hydroxyl, =O, =S, -NH2, -NO2, -CN, -COOH, -OC 1-3 Alkyl, -C 1-3 Alkyl, and C 1-3 Alkylene hydroxyl group, represents the junction site; When Formula 1 is Formula 2B: R 1c , R 2c , and R 5c Each independently selected from hydrogen, F, Cl, Br, I, C 1-3 Alkyl, -NO2, -CN, -OC 1-3 Alkyl; or R 1c and R 2c Together with the carbon atom to which it is attached, it forms a saturated or unsaturated 5-6 membered heterocyclic group containing 2 O atoms; and / or R 6c -CH2-NH2, -CH2-NH-C(O)-R 4c 、-CH2-NH-C(O)OR 4c , or C4H8OH; preferably, R 6c is -CH2-NH2, or -(CH2)4-OH; and / or R, R a , R b ,R 4c Each occurrence is independently hydrogen, F, Cl, Br, I, CH2OH, or C2H4OH; When Formula 1 is Formula 2C: R 1A and R 2A are independently selected from halogen, amino, -OR' and C 1-6 Alkyl; further preferably, R 1A and R 2A Each independently selected from fluorine, chlorine, amino, -OC 1-6 Alkyl and C 1-6 Alkyl; and / or R 3A Each occurrence is independently selected from hydrogen, -C(O)R', -C(S)R', -C(S)C(R a1 )(R b1 )OH、-C(O)CH2N(R a1 )(R b1 )、-CO2R'、-C(O)C(R a1 )(R b1 )OH, -C(O)-C 3-6 Cycloalkylene-OH, -C(O)-C 1-6 Alkylene -OH, -C(O)-C 3-6 Cycloalkylene-C 1-6 Alkylene -OH, -C(O)OC 1-6 Alkylene-OH, and C optionally substituted by R' 1-6 Alkyl, C 1-6 Alkenyl and C 1-6 Alkynyl; and / or Preferably, R 4A Selected from hydrogen, or C 1-6 Alkyl; and / or R', R a1 , and R b1 Each occurrence is independently hydrogen, fluorine, chlorine, -OH, C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, 3, 4, 5 or 6-membered cycloalkyl or 3, 4, 5 or 6-membered heterocyclyl containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, wherein the C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 The alkynyl, 3, 4, 5 or 6-membered cycloalkyl, 3, 4, 5 or 6-membered heterocyclyl is unsubstituted or substituted by 1 or 2 independently selected from fluorine, chlorine, -NO2, -CN, -OH, -NH2, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, and hydroxyl C 1-6 The alkylene group is substituted with a substituent.

4. The compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from: Wherein R3 is selected from the following groups: in Indicates the connection site.

5. A compound as shown in Formula 2 or a pharmaceutically acceptable salt thereof, in, R1 and R2 are each independently selected from a hydrogen atom, a halogen group, a C 1-8 Alkyl, C 1-8 Haloalkyl; A is selected from an oxygen atom or a sulfur atom; B is selected from -C(O)- or -P(O)(OH)-; R3 is selected from -(C(R 3a )(R 3b )) m -R 3c , When -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m -The n methylene units are each independently -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -O-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SO2N(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N- or -N=CH-, when R3 is When , W1 and W2 are fused together; R4 and R8 are single bonds or are independently selected from -O-, C 1-8 Alkylene, C 1-8 Haloalkylene, -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SO2N(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N- or -N=CH-; Ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from a 6-10 membered aryl group, a 5-10 membered heteroaryl group, a 3-10 membered cycloalkyl group, or a 3-10 membered heterocyclyl group; When ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from a cycloalkyl group or a heterocyclic group, the cycloalkyl group or the heterocyclic group is replaced by one or more R w1 , R w2 , R w3 , R w4 , R w5 Replacement, R w1 , R w2 , R w3 , R w4 , or R w5 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic groups; When ring W1, ring W2, ring W3, ring W4, and ring W5 are each independently selected from an aryl group or a heteroaryl group, the aryl group or the heteroaryl group is replaced by one or more R w1 , R w2 , or R w3 , R w4 , or R w5 Replacement, R w1 , R w2 , R w3 , R w4 , or R w5 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic groups; R6 is selected from hydrogen atom, halogen, C 1-8 Alkyl, C 1-8 Preferably, R6 is selected from a hydrogen atom; R7 is selected from C 1-8 Alkylene N(R a )(R b ), C 1-8 Haloalkylene N(R a )(R b ), halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Preferably, R7 is selected from C 1-8 Alkylene N(R a )(R b ), or C 1-8 Haloalkylene N(R a )(R b ); Preferably, R7 is selected from -CH2N(R a )(R b ); Or R6 and R7 can form a saturated or unsaturated 5-10 membered cycloalkyl or a 6-10 membered heterocyclic group containing 1-3 heteroatoms independently selected from N, O, and S, wherein the cycloalkyl or heterocyclic group is substituted by one or more independently selected from halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Preferably, R6 and R7 form a 6-membered cycloalkyl with the carbon atom to which they are attached, the cycloalkyl being -N(R a )(R b )replace; R 3a , R 3b , R 3c , R5 is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic groups; R, R a , R b Each occurrence is independently hydrogen, protium, deuterium, tritium, halide, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -C(O)CH2OH, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, or C 1-6 Aliphatic groups; m and n are integers greater than or equal to 0; When R3 is -(C(R 3a )(R 3b )) m -R 3c ,-(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m - the n methylene units are each independently replaced by -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -O-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SO2N(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N- or -N=CH-; Preferably, R3 is -(CH2) a (CH2CH2O) b (CH3) c , or C optionally substituted by R 1-8 Aliphatic group, m = a + 3 * b + c, b is not 0, a and b are integers greater than 0; when R3 is -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m -n methylene units are each independently replaced by -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -O-, -S-; wherein ring W1, ring W2 and ring W3 are each independently selected from 6-10 membered aryl, 5-8 membered heteroaryl, 4-8 membered cycloalkyl or 4-8 membered heterocyclyl; preferably, ring W1 is selected from phenyl or 4-8 membered heterocyclyl, ring W2 is selected from phenyl or 4-8 membered heterocyclyl, and ring W3 is selected from phenyl or 5-8 membered heteroaryl; preferably, ring W1 is phenyl or oxolanyl, Ring W2 is phenyl or oxolanyl, and ring W3 is phenyl or pyridyl; When ring W1, ring W2 or ring W3 is selected from cycloalkyl or heterocyclic group, the cycloalkyl or The heterocyclic group is replaced by one or more R w1 , R w2 or R w3 Replacement, R w1 , R w2 or R w3 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 When ring W1, ring W2 or ring W3 is selected from 6-10 membered aryl or 5-8 membered heteroaryl, the aryl or heteroaryl is replaced by one or more R w1 , R w2 or R w3 Replacement, R w1 , R w2 or R w3 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic group; when R3 is -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m -n methylene units are each independently replaced by -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -O-, -S-; wherein ring W1 is selected from 6-10 membered aryl, 4-8 membered cycloalkyl, 4-8 membered heterocyclyl, 5-8 membered heteroaryl, when ring W1 is selected from 4-8 membered cycloalkyl or 4-8 membered heterocyclyl, the cycloalkyl or heterocyclyl is replaced by one or more R w1 Replacement, R w1 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, -(=O), -(=S), -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 When ring W1 is selected from 6-10 membered aryl or 5-8 membered heteroaryl, the aryl or heteroaryl is replaced by one or more R w1 Replacement, R w1 Every time it appears are each independently selected from hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic groups; When R3 is -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m -n methylene units are each independently replaced by -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -O-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SO2N(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N-, or -N=CH-; Preferably, R3 is m1 is an integer from 1 to 6, and ring W1 is selected from 6-10 membered aryl, 4-8 membered cycloalkyl, 4-8 membered saturated or unsaturated heterocyclic group containing 1-3 heteroatoms selected from N, O, and S, and 5-8 membered heteroaryl containing 1-3 heteroatoms selected from N, O, and S. When ring W1 is selected from 4-8 membered cycloalkyl or 4-8 membered heterocyclic group, the cycloalkyl or heterocyclic group is replaced by one or more R w1 Replacement, R w1 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic groups; When ring W1 is selected from 6-10 membered aryl or 5-8 membered heteroaryl, the phenyl or 5-8 membered heteroaryl is replaced by one or more R w1 Replacement, R w1 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, -NO2, -CN, -OR, -SR, -N(R a )(R b )、-C(O)R、 -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic groups; Preferably, when the ring W1 is composed of one or more R w1 When the ring W1 is selected from a 4-8 membered heterocyclic group, it contains 1, 2 or 3 heteroatoms, and each occurrence of the heteroatoms is independently selected from N, O or S; preferably, the ring W1 is Ring W1 can be one or more R w1 Replacement, R w1 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic groups; When R3 is -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m -, wherein the n methylene units are each independently replaced by -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -O-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SO2N(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N- or -N=CH-; wherein ring W1 and ring W2 are each independently selected from substituted or unsubstituted 6-10 membered aryl, 5-8 membered heteroaryl, 4-8 membered cycloalkyl or 4-8 membered heterocyclyl; preferably, ring W1 is ... w1 substituted phenyl or 4-8 membered heterocyclic group, ring W2 is selected from w2 substituted 4-8 membered heterocyclic group; preferably, ring W1 is substituted by one or more R w1 Substituted benzene ring or oxolane ring, Ring W2 is selected from w2 Replaced When ring W1 or ring W2 is selected from a cycloalkyl group or a heterocyclic group, the cycloalkyl group or the heterocyclic group is w1 or R w2 Replacement, R w1 or R w2 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic groups; When ring W1 or ring W2 is selected from a 6-10 membered aryl group or a 5-8 membered heteroaryl group, the aryl or heteroaryl group is replaced by one or more R w1 or R w2 Replacement, R w1 or R w2 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic group, R4 is a single bond or selected from -O-, C 1-8 Alkylene, C 1-8 Haloalkylene, -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SO2N(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N- or -N=CH-; When R3 is -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m -The n methylene units are each independently -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -O-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SO2N(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N- or -N=CH-; Ring W1 and Ring W2 are each independently selected from a 6-10 membered aryl group, a 5-8 membered heteroaryl group, a 4-8 membered cycloalkyl group or a 4-8 membered heterocyclic group; Preferably, Ring W1 and Ring W2 are fused to form a fused ring structure selected from any of the following When ring W1 or ring W2 is a 4-8 membered cycloalkyl group or a 4-8 membered heterocyclic group, it is w1 or R w2 Replacement, R w1 or R w2 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic groups; When ring W1 or ring W2 is a 6-10 membered aryl group or a 5-8 membered heteroaryl group, the aryl group or heteroaryl group is replaced by one or more R w1 or R w2 Replacement, R w1 or R w2 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic groups; When R3 is -(C(R 3a )(R 3b )) m - contains a methylene unit, the -(C(R 3a )(R 3b )) m -n methylene units are each independently replaced by -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -O-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SO2N(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N- or -N=CH-; Rings W1, W2, W3 and W4 are sequentially fused and connected, and Rings W1 and W4 are each independently selected from a 4-8 membered heterocyclic group, which is replaced by one or more R w1 or R w4 Replacement, R w1 or R w4 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic group; Ring W5 and Ring W2 are 6-10 membered aryl or 5-8 membered heteroaryl, the aryl or heteroaryl being separated by one or more R w2 Replacement, R w2 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic group; Ring W3 is a 4-8 membered cycloalkyl group, which is w3 Replacement, R w3 Each occurrence is independently selected from hydrogen, protium, deuterium, tritium, halo, =O, =S, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b ), -SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic group; R4 is a single bond or selected from -O-, C 1-8 Alkylene, C 1-8 Haloalkylene, -N(R5)C(O)-, -C(O)N(R5)-, -C(O)-, -OC(O)-, -C(O)O-, -NR5-, -S-, -SO-, -SO2-, -P(R5)-, -P(=O)(R5)-, -N(R5)SO2-, -SO2N(R5)-, -C(=S)-, -C(=NR5)-, -N=N-, -CH=N- or -N=CH-; m is an integer from 1 to 20; n is an integer from 0 to 8.

6. A compound as shown in formula 1C or a pharmaceutically acceptable salt thereof, in, q is 1, 2, or 3; R 1c , R 2c , R 5c Each is independently selected from hydrogen, halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R, -S(O)2R, -C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic groups; or R 1c and R 2c Together with the carbon atom to which it is attached, it forms a saturated or unsaturated 5-10-membered cycloalkyl or a 6-10-membered heterocyclic group containing 1-3 heteroatoms independently selected from N, O, and S, wherein the cycloalkyl or heterocyclic group is substituted by one or more independently selected from halogen, -NO2, -CN, -OR, -SR, -N(R a )(R b ), -C(O)R, -CO2R, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)R、-S(O)2R、-C(O)N(R a )(R b )、-SO2N(R a )(R b ), -OC(O)R, -N(R)SO2R, or C optionally substituted by R 1-6 Aliphatic group substitution; R 6c C 1-8 Alkyl hydroxyl, C 1-8 Alkylamino, -C(R a )(R b )-N(R a )-R 4c ,-C(R a )(R b )-N(R a )-C(O)-R 4c , or -C(R a )(R b )-N(R a )-C(O)OR 4c ; R, R a , R b ,R 4c Each occurrence is independently hydrogen, protium, deuterium, tritium, halide, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 aliphatic group, 3-8 membered cycloalkyl group, 3-8 membered heterocyclyl group, 6-10 membered aryl group or 5-10 membered heteroaryl group; said C 1-6 The aliphatic group, 3-8 membered cycloalkyl group, 3-8 membered heterocyclyl group, 6-10 membered aryl group or 5-10 membered heteroaryl group is unsubstituted or substituted by one or more substituted alkyl groups independently selected from halo, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C ... 1-6 Aliphatic group, C 1-6 Alkylene hydroxy substituted.

7. The compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

8. A ligand-drug conjugate having a structure as shown in Formula 3 or a pharmaceutically acceptable salt thereof, in, Ab is the ligand, L is the linker moiety, and D is the drug moiety; n is any integer or decimal from 1 to 15; preferably, n is any integer or decimal from 1 to 13; preferably, n is any integer or decimal from 3 to 10; The drug portion D is any of the following structures, in, R1, R2, R3, R6, R7, A, B, R 1c , R 2c , R 5c , R 1A , R 2A , R 3A , R 4A , and q as defined in any one of claims 2, 3, 4, 5, 6, or 7; R 6A Selected from hydrogen, -CH(CH3)N(R 3A )(R 4A ) and -CH2N(R 3A )(R 4A ); R 5A is hydrogen or C 1-6 alkyl; R 1s , R 2s , R 7s , and R 7ss Each independently is C 1-8 Alkylene NR-, C 1-8 Preferably, R 1s , R 2s , R 7s , and R 7ss Each independently represents -NR-, -NR-CH2-, or -O-CH2C(O)NR-; preferably R 1s , R 2s , R 7s , and R 7ss Each independently is -NH-, -NH-CH2-, or -O-CH2C(O)NH-; R 6bs Each occurrence is independently selected from -CH2N(R a1 )-、-CH2N(R a1 )-C 2-5 Alkylene-O-, -CH2N(R a1 )C(O)C(R a1 )(R b1 )O-、-CH2N(R a1 )C(S)C(R a1 )(R b1 )O-、-CH2N(R a1 )C(O)-C 3-8 Cycloalkylene-O-, -CH2N(R a1 )C(O)-C 2-5 Alkylene-O-, -CH2N(R a1 )C(O)C(R a1 )(R b1 )N(R a1 )-、-CH2N(R a1 )C(O)-C 3-8 Cycloalkylene-C(R a1 )(R b1 )-O-、-C 2-5 Alkylene-O-, -CH2N(R a1 )C(O)OC 2-5 Alkylene-O-; Where R, R a1 , and R b1 Each occurrence is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, 3, 4, 5, 6, 7 or 8 membered cycloalkyl, 3-8 membered heterocyclyl containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, 6-10 membered aryl, or 5-10 membered heteroaryl containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, wherein the C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 The alkynyl, 3-, 4-, 5-, 6-, 7-, or 8-membered cycloalkyl, 3- to 8-membered heterocyclyl, 6- to 10-membered aryl, or 5- to 10-membered heteroaryl is unsubstituted or substituted with one or more substituted alkyl radicals independently selected from halogen, -NO2, -CN, -OH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, C ... 1-6 Alkyl, C 2-6 Alkenyl or C 1-6 Alkynyl, and hydroxyl C 1-6 The alkylene group is substituted with a substituent; and R 6aa Selected from -C 1-8 Alkylene N(R a2 )-、-CH2N(R a2 )C(O)C(R a2 )(R b2 )O-、-CH2N(R a2 )C(O)-C 3-8 Cycloalkylene-O-, -C 2-5 Alkylene-O-, -CH2N(R a2 )C(O)OC 2-5 Alkylene-O-; preferably R 6aa is -CH2-NH2, or -(CH2)4-O-; Among them, R a2 and R b2 Each occurrence is independently hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, or C 1-6 Preferably, R a2 and R b2 Each occurrence is independently hydrogen, F, Cl, Br, I, -CH2OH, or -C2H4OH.

9. The ligand-drug conjugate or a pharmaceutically acceptable salt thereof according to claim 8, wherein the drug moiety D is selected from any of the following structures: in, Indicates the connection site.

10. The ligand-drug conjugate according to claim 8 or 9, or a pharmaceutically acceptable salt thereof, The connecting part L is L 1 -L2 -L 3 -L 4 ; L 1 Selected from -(succinimidyl-3-yl-N)-, -(succinimidyl-3-yl-N)-WC(=O)-, or in, W is selected from C 1-10 Alkylene, C 1-10 Alkylene-cycloalkylene, C 1-10 Heteroalkylene, C 1-10 Alkylene-cycloheteroalkylene, or C 1-10 Heteroalkylene-cycloalkylene, preferably W is C 1-8 Alkylene, C 1-8 Alkylene-cycloalkylene or C 1-8 The heteroalkylene group comprises 1 to 3 heteroatoms independently selected from N, O or S, wherein the alkylene group, cycloalkylene group and heteroalkylene group are unsubstituted or each independently optionally further substituted by one or more substituents selected from halogen, hydroxyl, -CN, amino, alkyl, haloalkyl, deuterated alkyl, alkoxy and cycloalkyl, preferably, the alkylene group, cycloalkylene group and heteroalkylene group are unsubstituted or each independently optionally further substituted by one or more substituents selected from halogen, hydroxyl, -CN, amino, C 1-6 Alkyl, C 1-6 Haloalkyl, deuterated C 1-6 Alkyl, C 1-6 Alkoxy and C 5-8 More preferably, the alkylene, cycloalkylene and heteroalkylene are unsubstituted or optionally substituted by halogen, -OH, -CN, C 1-3 Alkyl, and -OC 1-3 Alkyl is substituted with a substituent, preferably, the alkylene, cycloalkylene and heteroalkylene are unsubstituted or optionally substituted with a substituent selected from Cl, Br, F, -OH, -CN, methyl, and -OCH3; X is selected from a single bond, C 1-10 Alkylene, C 1-10 Alkylene-cycloalkylene, C 1-10 Heteroalkylene, C 1-10 Alkylene-cycloheteroalkylene, or C 1-10 Heteroalkylene-cycloalkylene, preferably X is selected from a single bond, C 1-8 Alkylene, C 1-8 Alkylene-cycloalkylene or C 1-8 heteroalkylene; L 2 Choice-(CH2CH2O) r CH2CH2C(=O)-, -(CH2CH2O) r C(=O)-, -NR 1L (CH2CH2O) r C(=O)-, -(CH2CH2O) r CH2C(=O)-, -NR 1L (CH2CH2O) r CH2CH2C(=O)-, -NR 1L (CH2CH2O) r CH2C(=O)-, -NR 1L CH2-Ar 1 -(CH2CH2O) r CH2CH2NR 1L C(=O)CH2OCH2C(=O)-, -NR 1L (CH2CH2O) r CH2-Ar 1 -(CH2CH2O) r CH2CH2C(=O)-, -S(CH2) r C(=O)-, -O-(CH2CH2O) r CH2-Ar 1 -(CH2CH2O) r CH2CH2C(=O)-, or a single bond, wherein r is an integer of 12, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, n17 is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 at each occurrence; j is independently selected from 0, 1, 2, 3, 4, 5, 6 at each occurrence; preferably r is an integer of 1, 2, 3, 4, 5, 6, 7, or 8; Ar 1 is selected from 6-10 membered aryl, 5-10 membered heteroaryl containing 1-3 heteroatoms independently selected from N, O, P and S, 3-10 membered cycloalkyl or 3-10 membered heterocyclic group containing 1-3 heteroatoms independently selected from N, O, P and S; preferably, Ar 1 Selected from L 3 is a peptide residue consisting of 2 to 7 amino acids, preferably L 3 is a peptide residue consisting of 2, 3, 4, 5 or 6 amino acids, wherein the amino acids are unsubstituted or optionally further substituted by halogen, hydroxyl, -CN, The alkyl radical is substituted with one or more substituents selected from amino, alkyl, haloalkyl, deuterated alkyl, alkoxy and cycloalkyl, and is preferably further substituted with one or more substituents selected from halogen, hydroxy, -CN, amino, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Alkoxy and C 5-8 The cycloalkyl group is substituted with one or more substituents; L 4 selected from -NR 2L (CR 3L R 4L ) t -Z-(CR 3L R 4L ) t -C(=O)-, -NR 2L (CR 3L R 4L ) t -, -NR 2L (CR 3L R 4L ) t -Z-(CR 3L R 4L ) t -Z-C(=O)-, -NR2-Ar 2 -(CR 3L R 4L ) t -ZC(=O)-, or a single bond, wherein t is independently an integer of 0, 1, 2, 3, 4, 5, or 6 at each occurrence; Z is independently a single bond, O, S, or -NH- at each occurrence; Ar 2 is an arylene or heteroarylene group, preferably selected from a 6-membered arylene group or a 5-8-membered heteroarylene group, wherein the heteroarylene group contains 1, 2 or 3 heteroatoms, wherein the heteroatoms are independently selected from N, O and S; the arylene or heteroarylene group is unsubstituted or optionally selected from H, halogen, -OH, -CN, C 1-6 Alkyl, -OC 1-6 Alkyl, -NH2, -NH(C 1-6 alkyl), and -N(C 1-6 Alkyl)2 is substituted by a substituent; R 1L and R 2L are the same or different and are independently selected at each occurrence from hydrogen atoms, halogens, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl and -C 1-6 Alkylene-OH; R 3L and R 4L are the same or different and are independently selected at each occurrence from hydrogen atoms, halogens, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl and -C 1-6 Alkylene-OH; and L 1 The end is connected to the ligand, L 4 The end is connected to the drug portion.

11. The ligand-drug conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 8 to 10, wherein L 3 is a peptide residue consisting of 2 to 6 amino acids selected from glycine, phenylalanine, alanine, valine, lysine, citrulline, serine, glutamic acid, and aspartic acid; preferably a dipeptide residue, a tripeptide residue, or a tetrapeptide residue selected from alanine, phenylalanine, glycine, lysine, and citrulline; preferably, L 3 is a peptide residue selected from the group consisting of glycine-phenylalanine-glycine, alanine-alanine-alanine-glycine, alanine-alanine-alanine, glycine-glycine-phenylalanine-glycine, valine-citrulline, and valine-alanine; in, The peptide residue is unsubstituted or optionally further substituted by one or more substituents selected from halogen, hydroxyl, -CN, amino, alkyl, haloalkyl, deuterated alkyl, alkoxy and cycloalkyl, preferably, optionally further substituted by one or more substituents selected from halogen, hydroxyl, -CN, amino, and C 1-6 Alkyl substituents are substituted.

12. The ligand-drug conjugate according to any one of claims 8 to 11 or a pharmaceutically acceptable salt thereof, Wherein the linking part L is selected from: in Indicates the connection site.

13. The ligand-drug conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 8 to 12, wherein the ligand-drug conjugate is selected from the following structures: in, n is an integer or decimal from 1 to 10; preferably, n is an integer or decimal from 3 to 8; Ab is a ligand.

14. A compound as shown in Formula 5 or a pharmaceutically acceptable salt thereof, L j -L2-L3–L4-D (Formula 5); Wherein -L2-, -L3-, -L4-, -D are as defined in any one of claims 8 to 13; L j Selected from and W, X are as defined in any one of claims 8-13.

15. The compound according to claim 11, 16. The ligand-drug conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 8 to 13, wherein the Ab is an antibody or an antigen-binding fragment thereof or a polypeptide, wherein the antibody is selected from a chimeric antibody, a humanized antibody and a fully human antibody; Preferably, the antibody or its antigen-binding fragment is selected from anti-TROP-2 antibody, anti-HER2 (ErbB2) antibody, anti-NECTIN4 antibody, anti-EGFR antibody, anti-B7-H3 antibody, anti-c-Met antibody, anti-HER3 (ErbB3) antibody, anti-HER4 (ErbB4) antibody, anti-LIV-1 antibody, anti-ROR1 antibody, anti-CD20 antibody, anti-CD22 antibody, anti-CD30 antibody, anti-CD33 antibody, anti-CD44 antibody, anti-CD56 antibody, anti-CD70 antibody, anti-CD73 antibody, anti-CD105 antibody, anti-CEA antibody, anti-A33 antibody, anti-Cripto antibody, anti-EphA2 antibody, anti-G250 antibody, anti-MUCl antibody, anti-Lewis Y antibody, anti-VEGFR antibody, anti-GPNMB antibody, anti-Integrin antibody, anti-PSMA antibody, anti-Tenascin-C antibody, anti-SLC44A4 antibody, anti-Mesothelin antibody or its antigen-binding fragment Fragment; Preferably, the antibody or antigen-binding fragment thereof is an anti-TROP-2 antibody, an anti-NECTIN4 antibody, an anti-B7-H3 antibody, an anti-HER2 (ErbB2) antibody, an anti-HER3 (ErbB3) antibody, an anti-LIV-1 antibody, an anti-ROR1 antibody or an antigen-binding fragment thereof; Preferably, the antibody or antigen-binding fragment thereof is an anti-HER2 (ErbB2) antibody, an anti-NECTIN4 antibody, an anti-B7-H3 antibody or an antigen-binding fragment thereof; Preferably, the antibody or antigen-binding fragment thereof is trastuzumab, ifinatamab monoclonal antibody or PADCEV monoclonal antibody.

17. A pharmaceutical composition comprising a therapeutically effective amount of the compound or ligand-drug conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 16, and a pharmaceutically acceptable carrier.

18. Use of the compound or ligand-drug conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 16, or the pharmaceutical composition according to claim 17 in the preparation of a drug for treating or preventing tumors; Preferably, the tumor is a cancer associated with B7-H3 expression, HER2 expression, or NECTIN4 expression; Preferably, the cancer is selected from breast cancer, gastric cancer, melanoma and lung cancer.

19. A method for preventing or treating tumors, comprising administering an effective amount of the compound or ligand-drug conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 16, or the pharmaceutical composition according to claim 17 to a subject in need thereof; Preferably, the tumor is a cancer associated with B7-H3 expression, HER2 expression, or NECTIN4 expression; Preferably, the cancer is selected from breast cancer, gastric cancer, melanoma and lung cancer.

20. The compound or ligand-drug conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 16, or the pharmaceutical composition according to claim 17, for use in preventing or treating tumors; Preferably, the tumor is a cancer associated with B7-H3 expression, HER2 expression or NECTIN4 expression; preferably, the cancer is selected from breast cancer, gastric cancer, melanoma and lung cancer.

21. The use of a compound according to any one of claims 1 to 7, characterized in that Used as a toxin in antibody-drug conjugates to prepare antibody-drug conjugates.

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