Ecteinascidins derivative, ligand-drug conjugate and pharmaceutical use thereof

Abstract

Provided are a ecteinascidins derivative, a ligand-drug conjugate, a pharmaceutical composition containing the ecteinascidins derivative or conjugate, and a corresponding pharmaceutical use thereof.

Description

Tunicamycin derivatives, ligand-drug conjugates and medical uses thereof

[0001] This application claims priority to:

[0002] Chinese patent CN202411124134.4 filed on August 16, 2024,

[0003] Chinese patent CN 202411240899.4 filed on September 5, 2024,

[0004] Chinese patent CN202411466062.1 filed on October 21, 2024,

[0005] Chinese patent CN202510338506.1 filed on March 21, 2025. TECHNICAL FIELD

[0006] The present application relates to the field of medicine, in particular to tunicamycin derivatives, ligand-drug conjugates and medical uses thereof in treating or preventing proliferative diseases. BACKGROUND

[0007] Ecteinascidins are a class of natural products. In 1990, the KL Rinehart laboratory at the University of Illinois isolated six ecteinascidins from Caribbean tunicates, one of which is trabectedin. In 2015, trabectedin was approved by FDA for the treatment of patients with unresectable or metastatic liposarcoma or leiomyosarcoma who have been previously treated with an anthracycline. In addition to soft tissue sarcoma, trabectedin has also been approved by regulatory agencies in Europe, Canada and other regions for the treatment of platinum-sensitive recurrent ovarian cancer. Subsequent studies have found that the mechanism of action of trabectedin is relatively unique. Unlike the mechanism of action of paclitaxel and eribulin in inhibiting microtubules, trabectedin can prevent cell division by binding to DNA: two fused tetrahydroisoquinoline rings can interact with and covalently bind to specific sequences in the DNA minor groove, causing DNA double helix abnormalities and interfering with the normal function of XPG protein in the TC-NER system, thereby inducing DNA double-strand breaks and causing cell death. Rubitecan is a derivative of ecteinascidin developed by PharmaMar, an innovative drug for the treatment of small cell lung cancer. The clinical data of the drug analyzed 105 cases of extensive SCLC patients who had previously progressed after first-line chemotherapy, with rubitecan 3.2 mg / m2, 1 hour intravenous infusion, 3 weeks once. The results showed that the objective response rate of second-line treatment was 35.2%, and the disease control rate was 68.6%. The median response duration was 5.3 months, the median survival of patients was 9.3 months, and the 1-year survival rate was 34.2%. The most common 1 / 2 grade adverse reactions included fatigue (51.4%), nausea (32.4%), decreased appetite (21.0%), vomiting (18.1%), diarrhea (12.4%), constipation (9.5%), and neutropenia (5.7%). 3 / 4 serious adverse reactions included neutropenia, anemia, fatigue, thrombocytopenia, febrile neutropenia, pneumonia, elevated alanine aminotransferase levels, skin ulcers and diarrhea. Therefore, it is necessary to find better ecteinascidin drugs.

[0008] Ligand-drug conjugates (ADCs) as a new type of targeted drugs are generally composed of three parts: antibodies or antibody-like ligands, small molecule drugs, and connecting parts that couple ligands and drugs. Antibody-drug conjugates use the specific recognition of antibodies to antigens to transport drug molecules to the vicinity of target cells and effectively release drug molecules to achieve therapeutic purposes. In 2000, Mylotarg, an ADC drug from Pfizer, was first marketed, and the field of ADCs with great potential and challenges has since entered the public eye. In recent years, the pharmaceutical market has ushered in a new round of ADC research and development boom, and currently there are 13 ADC drugs on the market worldwide.

[0009] It is necessary and urgent to explore and utilize better ascidian derivatives, improve the safety and effectiveness of small-molecule compounds against tumors in the application of ADC drugs, and obtain anti-tumor drugs with excellent efficacy. SUMMARY

[0010] In order to solve the problems existing in the prior art, the present application provides a compound as shown in formula I-1, or an isomer, an isotopic derivative, a polymorph, a prodrug, a pharmaceutically acceptable salt or a solvate thereof,

[0011] wherein R1 and R2 are each independently selected from H, halogen, hydroxyl, C(=O)R0, C(=O)OR0, -OC(=O)OR0, -OC(=O)R0, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, and substituted or unsubstituted C1-C6 haloalkyl; the C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, and C1-C6 haloalkyl are optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6 alkyl;

[0012] Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, and Y9 are each independently selected from H, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, OH, NO2, NH2, SH, CN, or halogen;

[0013] X1 is each occurrence independently selected from halogen, alkyl, haloalkyl, OR x , CN, (=O), and H; wherein R x is selected from H, alkyl, haloalkyl, alkenyl, alkynyl, OH, OR0, SR0, SOR0, and SO2R0;

[0014] R3 is selected from CR 3a R 3b ;

[0015] R 3a is H, alkyl, alkoxy, OH, NO2, NH2, SH, CN, or halogen;

[0016] R 3b is selected from

[0017] R 3aa is

[0018] R 3ab is selected from OH, and N(R00 )2;

[0019] R 3b is not

[0020] R4is selected from C(R 4a )2;

[0021] R 4a each occurrence is independently selected from H, alkyl, alkoxy, OH, NO2, NH2, SH, CN, and halogen;

[0022] or R3is CR 3a , R4is CR 4a , and R3and R4together with the atoms to which they are attached form

[0023] Q1is independently selected at each occurrence from NR 00 , C(R 00 )2, O, and S;

[0024] Q2is independently selected at each occurrence from a single bond, NR 00 , C(R 00 )2, O, and S;

[0025] R a1 and R a2 are each independently selected from H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl;

[0026] R5and R6are each independently selected from H, halogen, C(=O)R0, C(=O)OR0, substituted or unsubstituted C1-C6alkyl, C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl; said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, and C2-C6alkynyl are optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6alkyl;

[0027] Q4is CR0or N, Q3is CR0or N;

[0028] Q6is C;

[0029] Q5is NR0;

[0030] R c1 , R c2 , R c3 , R c4 , R c6 , R c7 , and R c8 are each independently selected from R c0 , H, OH, -NO2, -NH2, -SH, -CN, halogen, -C(=O)H, -C(=O)CH3, -CO2H, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, and substituted or unsubstituted C2-C6alkynyl; said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, and C2-C6alkynyl are optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C6alkyl;

[0031] R c5 is selected from R c0 , H, OH, -NO2, -NH2, -SH, -CN, halogen, -C(=O)H, -C(=O)CH3, -CO2H, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, and substituted or unsubstituted C2-C6alkynyl;

[0032] R c5 is not

[0033] R c9 is R c0 , and R c9 is not , and R6is H, R c9 is

[0034] R c0 is R ca -(R cb ) m0 -,

[0035] R ca is selected from

[0036] R cb are each independently selected at each occurrence from -C(O)-, -N(R0)-, -O-, -C(S)-, -SO2-, -C(R0)2-;

[0037] m0 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;

[0038] m1 is at each occurrence independently selected from 0, 1, 2, 3, 4, 5, 6, and 7;

[0039] m2 is at each occurrence independently selected from 1, 2, 3, 4, 5, and 6;

[0040] R3 is CR 3a , R4 is CR 4a , and R3 and R4 form, with the atoms to which they are attached, formula 1-F: when R c6 is H, R c5 is not H, methoxy, hydroxy, methyl, -NH-methyl, F, -NHCOCH2OH; or when R c5 is amino, R c6 is not methoxy;

[0041] X2, X3, X4, X5, and X6 are at each occurrence independently selected from H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, C1-C6alkoxy, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl are optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6alkyl;

[0042] R0 and R 00 are at each occurrence independently selected from H, OH, NO2, NH2, SH, CN, halogen, -C(=O)H, -C(=O)-C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl are optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6alkyl;

[0043] Cy is selected from the group consisting of 3-10 membered cycloalkylene, 3-10 membered heterocyclylene containing at least one heteroatom independently selected from N, O, and S, 6-10 membered arylene, and 5-10 membered heteroarylene containing at least one heteroatom independently selected from N, O, and S, said 3-10 membered cycloalkylene, 3-10 membered heterocyclylene, 6-10 membered arylene, and 5-10 membered heteroarylene optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, C 1-6 alkyl, C 1-6 alkoxy, hydroxyl, -NO2, -NH2, and -CN; and

[0044] n is at each occurrence independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10.

[0045] A second aspect of the present application provides a compound of Formula I:

[0046] wherein R1and R2are each independently selected from H, halogen, hydroxyl, C(=O)R0, C(=O)OR0, -OC(=O)OR0, -OC(=O)R0, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted C1-C6alkoxy, and substituted or unsubstituted C1-C6haloalkyl; in one embodiment, R1and R2are each independently selected from H, halogen, hydroxyl, C(=O)R0, C(=O)OR0, -OC(=O)OR0, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted C1-C6alkoxy, and substituted or unsubstituted C1-C6haloalkyl, said C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C1-C6alkoxy, and C1-C6haloalkyl optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; in one embodiment, R1is -OC(=O)CH3; and / or, R2is methyl;

[0047] Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, and Y9are each independently selected from H, C1-C6alkyl, C1-C6alkoxy, C1-C6haloalkyl, OH, NO2, NH2, SH, CN, or halogen; in one embodiment, Y1is OH, and / or, Y2is -OCH3, and / or Y3is CH3, and / or, Y4, Y5, Y6, Y7, Y8, and Y9are each independently H;

[0048] X1is at each occurrence independently selected from halogen, alkyl, haloalkyl, OR xH, alkyl, haloalkyl, alkenyl, alkynyl, OH, OR0, SR0, SOR0, and SO2R0; in one embodiment, X1is selected from the group consisting of hydroxyl and CN; x H, alkyl, haloalkyl, alkenyl, alkynyl, OH, OR0, SR0, SOR0, and SO2R0; in one embodiment, X1is selected from the group consisting of hydroxyl and CN;

[0049] R3is selected from the group consisting of CR 3a R 3b ;

[0050] R 3a is H, alkyl, alkoxy, OH, NO2, NH2, SH, CN, or halogen;

[0051] R 3b is selected from the group consisting of

[0052] R 3aa is

[0053] R 3ab is selected from the group consisting of OH, and N(R 00 )2;

[0054] R 3b is not

[0055] R4is selected from the group consisting of C(R 4a )2;

[0056] R 4a each occurrence is independently selected from the group consisting of H, alkyl, alkoxy, OH, NO2, NH2, SH, CN, and halogen;

[0057] or R3is CR 3a , R4is CR 4a , and R3and R4together with the atoms to which they are attached form

[0058] Q1and Q2are each independently selected from the group consisting of C(R 00 )2, O, and S;

[0059] R a1 , and R a2 are each independently selected from the group consisting of H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; in an embodiment, R a1 a2 each independently selected from the group consisting of H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, C1-C6alkoxy, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, C1-C6alkoxy, aryl, and heteroaryl optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C6alkyl;

[0060] when neither Q1nor Q2is R a1 and R a2 at least one of R when at least one of Q1and Q2is R a1 and R a2 each independently selected from the group consisting of H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, NH2, NHR0, N(R0)2, NHC(O)R0, CN, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; in an embodiment, R a1 and R a2 ​each independently selected from H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, C1-C6alkoxy, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, C1-C6alkoxy, aryl, and heteroaryl optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6alkyl;

[0061] R5and R6are each independently selected from H, halogen, C(=O)R0, C(=O)OR0, substituted or unsubstituted C1-C6alkyl, C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, and substituted or unsubstituted C2-C6alkynyl, said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, and C2-C6alkynyl optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6alkyl;

[0062] Q4is CR0or N, and Q3is CR0or N; in an embodiment, Q4is CR0or N, and Q3is CR0or N; and at least one of Q3and Q4is N; in a further embodiment, Q4is CH or N, and Q3is CH or N, at least one of Q3and Q4is N;

[0063] Q6is C;

[0064] Q5is NR0; in an embodiment, Q5is NH;

[0065] R c1 , R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , and R c8 are each independently selected from R c0H, OH, -N02, -NH2, -SH, -CN, halogen, -C(=0)H, -C(=0)CH3, -C02H, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, and substituted or unsubstituted C2-C6alkynyl; in an embodiment, R c1 c2 c3 c4 c5 c6 c7 c8 each R is independently selected from R c0 H, OH, -N02, -NH2, -SH, -CN, halogen, -C(=0)H, -C(=0)CH3, -C02H, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, and substituted or unsubstituted C2-C6alkynyl, said Ci-C6alkyl, Ci-C6alkoxy, C2-C6alkenyl, and C2-C6alkynyl being optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and Ci-C6alkyl;

[0066] R c5 c6 c7 c8 at least one of R c0 ; in an embodiment, one and only one of R c5 c6 c7 c8 is R c0 ;

[0067] R c9 is R c0 , and R c9 is not

[0068] R c0 is R ca -(R cb ) m0 -,

[0069] R ca is selected from

[0070] R cb each occurrence is independently selected from a single bond, methylene, -C(O)-, -N(R0)-, ​​​​​​​​​​​​​

[0071] m0 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;

[0072] m1 is at each occurrence independently selected from 0, 1, 2, 3, 4, 5, 6, and 7;

[0073] m2 is at each occurrence independently selected from 1, 2, 3, 4, 5, and 6;

[0074] R3 is CR 3a , R4 is CR 4a , and R3 and R4, together with the atoms to which they are attached, form when X1 is -CN, R c1 is not when X1 is -OH, R c1 is not In an embodiment, when R3 is CR 3a , R4 is CR 4a , and R3 and R4, together with the atoms to which they are attached, form , R c1 is not

[0075] X2, X3, X4, X5, and X6 are at each occurrence independently selected from H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, C1-C6alkoxy, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; in an embodiment, X2, X3, X4, X5, and X6 are at each occurrence independently selected from H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl being optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6alkyl;

[0076] R0 and R 00each occurrence is selected independently from H, OH, N02, NH2, SH, CN, halogen, -C(=0)H, -C(=0)-Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; in an embodiment, R0and R 00 each occurrence is selected independently from H, OH, N02, NH2, SH, CN, halogen, -C(=0)H, -C(=0)-Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; in an embodiment, R0and R 1-6 substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said Ci-C6alkyl, Ci-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl being optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and Ci-C6alkyl; in an embodiment, R0and R 00 each occurrence is selected independently from H, OH, N02, NH2, SH, CN, halogen, -C(=0)H, -C(=0)-Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; in an embodiment, R0and R 1-3 substituted or unsubstituted Ci-C3alkyl, substituted or unsubstituted C2-C3alkenyl, substituted or unsubstituted C2-C3alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said Ci-C3alkyl, Ci-C3alkoxy, C2-C3alkenyl, C2-C3alkynyl, aryl, and heteroaryl being optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and Ci-C3alkyl; in an embodiment, R0and R 00 each occurrence is selected independently from H, OH, F, Cl, Br, I, substituted or unsubstituted Ci-C3alkoxy, and substituted or unsubstituted Ci-C3alkyl, said Ci-C3alkyl, and Ci-C3alkoxy being optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and Ci-C3alkyl;

[0077] Cy is selected from 3-10 membered cycloalkylene, 3-10 membered heterocyclylene containing at least one heteroatom independently selected from N, O, and S, 6-10 membered arylene, and 5-10 membered heteroarylene containing at least one heteroatom independently selected from N, O, and S, said 3-10 membered cycloalkylene, 3-10 membered heterocyclylene, 6-10 membered arylene, and 5-10 membered heteroarylene being optionally substituted with a substituent selected from a deuterium atom, halogen, C 1-6 alkyl, C 1-6substituted with a substituent selected from the group consisting of deuterium, halogen, C1-C6alkyl, C1-C6alkoxy, hydroxyl, -NO2, -NH2, and -CN; and 1-6 C1-C6alkyl, C1-C6alkoxy, hydroxyl, -NO2, -NH2, and -CN; and 1-6 C1-C6alkyl, C1-C6alkoxy, hydroxyl, -NO2, -NH2, and -CN; and

[0078] n each occurrence is independently selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;

[0079] In one embodiment, R3is CR 3a , R4is CR 4a , and R3and R4, when taken together with the atoms to which they are attached, form a ring structure selected from the group consisting of:

[0080] In one embodiment, when R3is not connected to R4to form a ring, R 3b is

[0081] In one embodiment, wherein

[0082] R1is selected from the group consisting of H, F, Cl, Br, I, hydroxyl, -OC(=O)O-C1-C6alkyl, and C1-C6alkyl; in one embodiment, R1is selected from the group consisting of H and -OC(=O)O-C1-C3alkyl; in one embodiment, R1is -OC(=O)CH3; and / or,

[0083] R2is selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and C1-C6alkyl; in one embodiment, R2is selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and C1-C3alkyl; in one embodiment, R2is methyl; and / or

[0084] Y1is hydroxyl; and / or

[0085] Y2is selected from the group consisting of H, C1-C6alkyl, and C1-C6alkoxy; in one embodiment, Y2is selected from the group consisting of H, C1-C3alkyl, and C1-C3alkoxy; in one embodiment, Y2is C1-C3alkoxy; in one embodiment, Y2is -OCH3; and / or

[0086] Y3is selected from the group consisting of H, C1-C6alkyl, and C1-C6alkoxy; in one embodiment, Y3is selected from the group consisting of H, C1-C3alkyl, and C1-C3alkoxy; in one embodiment, Y3is C1-C3alkyl; in one embodiment, Y3is CH3; and / or

[0087] Y4, Y5, Y6, Y8, and Y9 are each independently selected from the group consisting of H, C1-C3 alkyl, and C1-C3 alkoxy; in an embodiment thereof, Y4, Y5, Y6, Y8, and Y9 are each independently H; and / or

[0088] Y7 is selected from the group consisting of H, C1-C6 alkyl, and C1-C6 alkoxy; in an embodiment thereof, Y7 is selected from the group consisting of H, C1-C3 alkyl, and C1-C3 alkoxy; in an embodiment thereof, Y7 is selected from the group consisting of H, and C1-C3 alkyl; Y7 is C1-C3 alkyl; in an embodiment thereof, Y7 is CH3; and / or

[0089] X1is at each occurrence independently selected from the group consisting of halogen, C1-C6 alkyl, C1-C6 haloalkyl, hydroxyl, C1-C6 alkoxy, CN, (=0), and H; in an embodiment thereof, X1is at each occurrence independently selected from the group consisting of F, Cl, Br, I, C1-C3 alkyl, C1-C3 haloalkyl, hydroxyl, C1-C3 alkoxy, CN, and H; in an embodiment thereof, X1is selected from the group consisting of hydroxyl and CN; and / or

[0090] R3is CR 3a , R4is CR 4a ; in an embodiment thereof, R3and R4are each independently selected from the group consisting of CH, C-F, C-Cl, C-Br, C-I, C-hydroxyl, and C-C1-C3 alkyl; in an embodiment thereof, R3and R4are each independently CH;

[0091] and R3and R4together with the atoms to which they are attached form and / or

[0092] R a1 and R a2 are each independently selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and C1-C6 alkyl; in an embodiment thereof, R a1 and R a2 are each independently selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and C1-C3 alkyl; in an embodiment thereof, R a1 and R a2 are each independently H; and / or

[0093] R5and R6are each independently selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and C1-C6 alkyl; in an embodiment thereof, R5and R6are each independently selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and C1-C3 alkyl; in an embodiment thereof, R5and R6are each independently H; and / or

[0094] Q4is CR0or N, Q3is CR0or N; and at least one of Q3and Q4is N; in a further embodiment, Q4is CH or N, Q3is CH or N, at least one of Q3and Q4is N; and / or

[0095] Q5is NH; and / or

[0096] Q6is C; and / or

[0097] Q2is selected from the group consisting of O, S, C(Ci-C6alkyl)2, C(Ci-C6alkyl)H, CH2, N(Ci-C6alkyl), and NH; in an embodiment, Q2is selected from the group consisting of O, S, C(Ci-C3alkyl)2, C(Ci-C3alkyl)H, CH2, N(Ci-C3alkyl), and NH; in an embodiment, Q2is selected from the group consisting of O, CH2, and NH; and / or

[0098] Q1is selected from the group consisting of O, S, C(Ci-C6alkyl)2, C(Ci-C6alkyl)H, CH2, N(Ci-C6alkyl), and NH; in an embodiment, Q1is selected from the group consisting of O, S, C(Ci-C3alkyl)2, C(Ci-C3alkyl)H, CH2, N(Ci-C3alkyl), and NH; in an embodiment, Q1is selected from the group consisting of O, CH2, and NH; in an embodiment, Q1is selected from the group consisting of O and NH;

[0099] R c1 , R c2 , R c3 , R c4 , R c6 , R c7 , and R c8 are each independently selected from the group consisting of R c0 , H, OH, -NO2, -NH2, -SH, -CN, halogen, -C(=O)H, -C(=O)CH3, -CO2H, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, and substituted or unsubstituted C2-C6alkynyl, said Ci-C6alkyl, Ci-C6alkoxy, C2-C6alkenyl, and C2-C6alkynyl optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and Ci-C6alkyl;

[0100] in an embodiment, R c6 is selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and Ci-C6alkyl; in an embodiment, R c6 is selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and Ci-C3alkyl; in an embodiment, R c6 is selected from the group consisting of H and F; and / or

[0101] In one embodiment, R c7 is selected from H, F, CI, Br, I, hydroxyl, and Ci-C6alkyl; in one embodiment, R c7 is selected from H, F, CI, Br, I, hydroxyl, and Ci-C3alkyl; in one embodiment, R c7 is H; and / or

[0102] In one embodiment, R c8 is selected from H, F, CI, Br, I, hydroxyl, and Ci-C6alkyl; in one embodiment, R c8 is selected from H, F, CI, Br, I, hydroxyl, and Ci-C3alkyl; in one embodiment, R c8 is H; and / or

[0103] R c5 is selected from R c0 , -NH2, and -(CH2) 1-4 -OH;

[0104] R c0 is R ca -(R cb ) m0 -,

[0105] R ca is selected from

[0106] R cb each occurrence is independently selected from methylene, -C(O)-, -O-, -N(R0)-, In one embodiment, R cb each occurrence is independently selected from methylene, -C(O)-, -N(R0)-,

[0107] m0is selected from 0, 1, 2, 3, 4, 5, and 6;

[0108] m1is each occurrence independently selected from 0, 1, and 2; in one embodiment, m1is each occurrence independently 1;

[0109] m2is each occurrence independently selected from 1, and 2; in one embodiment, m2is each occurrence independently 1;

[0110] In one embodiment, R c5 is selected from -NH2, -(CH2) m3 OH, -(CH2)m3 NHCH3, -(CH2) m3 NH2,

[0111] m1is each independently at each occurrence selected from 0, 1, and 2; in an embodiment thereof, m1is each independently at each occurrence 1;

[0112] m2is each independently at each occurrence selected from 1 and 2; in an embodiment thereof, m2is each independently at each occurrence 1;

[0113] m3is selected from 0, 1, 2, 3, 4, and 5;

[0114] m4is selected from 0, 1, 2, 3, 4, and 5; in an embodiment thereof, m4is selected from 0 and 1;

[0115] in an embodiment thereof, R c5 selected from -NH2, -(CH2) 1-4 -OH,

[0116] in an embodiment thereof, R c9 selected from and / or

[0117] X2, X3, X4, X5, and X6are each independently at each occurrence selected from H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6alkyl;

[0118] in an embodiment thereof, X2, X3, X4, and X5are each independently at each occurrence selected from H, F, Cl, Br, I, hydroxyl, and C1-C6alkyl; in an embodiment thereof, X2, X3, X4, and X5are each independently at each occurrence selected from H, F, Cl, Br, I, hydroxyl, and C1-C3alkyl; in an embodiment thereof, X2, X3, X4, and X5are each independently at each occurrence H; and / or

[0119] In an embodiment, X6is selected from the group consisting of H, F, Cl, Br, I, hydroxyl, C1-C6alkyl, -C1-C6alkyl-OH, and -C1-C6alkyl-NH2; in an embodiment, X6is selected from the group consisting of H, F, Cl, Br, I, hydroxyl, C1-C3alkyl, -C1-C3alkyl-OH, and -C1-C3alkyl-NH2; X6is selected from the group consisting of H, -CH2-OH, and -CH2-NH2; and / or

[0120] R0and R 00 each occurrence is independently selected from the group consisting of H, OH, NO2, NH2, SH, CN, F, Cl, Br, I, -C(=O)H, -C(=O)-C 1-6 alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; in an embodiment, R0and R 00 each occurrence is independently selected from the group consisting of H, OH, NO2, NH2, SH, CN, F, Cl, Br, I, -C(=O)H, -C(=O)-C 1-3 alkyl, substituted or unsubstituted C1-C3alkoxy, substituted or unsubstituted C1-C3alkyl, substituted or unsubstituted C2-C3alkenyl, substituted or unsubstituted C2-C3alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said C1-C3alkyl, C1-C3alkoxy, C2-C3alkenyl, C2-C3alkynyl, aryl, and heteroaryl optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C3alkyl; in an embodiment, R0and R 00 each occurrence is independently selected from the group consisting of H, OH, F, Cl, Br, I, substituted or unsubstituted C1-C3alkoxy, and substituted or unsubstituted C1-C3alkyl, said C1-C3alkyl, and C1-C3alkoxy optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C3alkyl; in an embodiment, R0each occurrence is independently selected from the group consisting of H and C1-C3alkyl; in an embodiment, R0each occurrence is independently selected from the group consisting of H and CH3; and / or

[0121] Cy is phenylene optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, C 1-6 alkyl, C 1-6substituted with substituents selected from the group consisting of alkyl, alkoxy, hydroxy, -NO2, -NH2, and -CN.

[0122] In one embodiment, R3is CR 3a , R4is CR 4a , and R3and R4, together with the atoms to which they are attached, form a ring selected from:

[0123] When R3is not connected to R4to form a ring, R 3b is

[0124] In one embodiment,

[0125] R 3a and R 4a are each independently selected from the group consisting of H, F, Cl, Br, I, hydroxy, and C1-C3 alkyl; in one embodiment, R 3a and R 4a are each independently H.

[0126] In one embodiment, the compound is

[0127] a1is selected from 0 and 1 ;

[0128] wherein X1is selected at each occurrence independently from CN and OH;

[0129] R c5 is selected from -NH2, -(CH2) m3 OH, -(CH2) m3 NHCH3, -(CH2) m3 NH2

[0130] m1is selected from 0, 1, 2, 3, 4, 5; in one embodiment, m1is selected from 0, 1, 2, and 3;

[0131] m2is selected from 1, 2, 3, 4, and 5; in one embodiment, m2is selected from 1, 2, and 3;

[0132] m3is selected from 0, 1, 2, 3, 4, and 5;

[0133] m4 is selected from 0, 1, 2, 3, 4, and 5; in an embodiment thereof, m4 is selected from 0 and 1;

[0134] R c6 is selected from H, OH, F, Cl, Br, I, and C1-C3 alkyl; in an embodiment thereof, R c6 is selected from H and F;

[0135] Q1and Q2are each independently selected from NR0, O, S, and C(R0)2; in an embodiment thereof, Q1is selected from NR0, O, and S; Q2is selected from C(R0)2, O, and S; in an embodiment thereof, Q1is selected from NH; Q2is selected from CH2, and O;

[0136] R0is selected from H, OH, F, Cl, Br, I, C1-C3 alkoxy, and C1-C3 alkyl; in an embodiment thereof, R0is each independently selected from H and C1-C3 alkyl at each occurrence thereof; more in an embodiment thereof, R0is each independently selected from H and CH3at each occurrence thereof;

[0137] X6is selected from H, -(CH2) m1 OH, and -(CH2) m1 NH2; in an embodiment thereof, X6is selected from H, -CH2-OH, and -CH2-NH2;

[0138] when R c5 is -(CH2) m3 NHCH3, -NH2, or -NHCOCH2OH, R c6 is selected from OH, F, Cl, Br, I, and C1-C3 alkyl; in an embodiment thereof, R c6 is F.

[0139] in an embodiment thereof, the compound is selected from:

[0140] A third aspect of the present application provides a drug linker conjugate as shown in Formula II, or an isomer, isotopic derivative, polymorph, prodrug, pharmaceutically acceptable salt or solvate thereof,

[0141] L0-D0(Formula II);

[0142] wherein,

[0143] L0is a linker precursor;

[0144] D0is a compound as described in any of the preceding at the R 3b , R a1 , R a2 , R c0 , R c1 , R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , Q1, or Q2 terminal group; in an embodiment, D0is a compound as described in any of the preceding at the R 3b , R a1 , R a2 , R c0 , R c1 , R c2 , R c3 , R c4 , R c5 , Q1, or Q2 terminal group; in an embodiment, D0is a compound as described in any of the preceding at the R 3b , R a1 , R a2 , R c0 , Q1, or Q2 terminal group; in an embodiment, D0is a compound as described in any of the preceding at the R c5 terminal group of Q1or a group formed by the loss of a hydrogen atom from the hydroxyl or amino group of the terminal group of Q2of formula I-C; the terminal fragment of the group after the loss of the hydrogen atom is selected from: wherein, represents the point of attachment of D0to L0after the loss of a hydrogen atom; represents the point of attachment of the terminal group to the rest of the compound;

[0145] R0and m1are as defined in any of the preceding; in an embodiment, m1is 1; in an embodiment, R0is selected from -CH3and H;

[0146] the linker precursor forms a linker upon attachment to the antibody.

[0147] The fourth aspect of the present application provides a drug linker conjugate as shown in formula II-1, or an isomer, isotopic derivative, polymorph, prodrug, pharmaceutically acceptable salt, or solvate thereof,

[0148] L0-D0(formula II-1);

[0149] wherein,

[0150] L0is a linker precursor;

[0151] D0is represented by Formula I-2,

[0152] wherein R1and R2are each independently selected from the group consisting of H, halogen, hydroxyl, C(=O)R0, C(=O)OR0, -OC(=O)OR0, -OC(=O)R0, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted C1-C6alkoxy, and substituted or unsubstituted C1-C6haloalkyl; in one embodiment, R1and R2are each independently selected from the group consisting of H, halogen, hydroxyl, C(=O)R0, C(=O)OR0, -OC(=O)OR0, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted C1-C6alkoxy, and substituted or unsubstituted C1-C6haloalkyl, said C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C1-C6alkoxy, and C1-C6haloalkyl optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C6alkyl;

[0153] Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, and Y9are each independently selected from the group consisting of H, C1-C6alkyl, C1-C6alkoxy, C1-C6haloalkyl, OH, NO2, NH2, SH, CN, or halogen;

[0154] X1is each occurrence, independently selected from the group consisting of halogen, alkyl, haloalkyl, OR x , CN, (=O), and H; wherein R x is selected from the group consisting of H, alkyl, haloalkyl, alkenyl, alkynyl, OH, OR0, SR0, SOR0, and SO2R0;

[0155] R3and R4are each independently selected from the group consisting of CH, C-F, C-Cl, C-Br, C-I, C-hydroxyl, and C-C1-C6alkyl; and R3and R4together with the atoms to which they are attached form

[0156] Q1is each occurrence, independently selected from the group consisting of NR 00 , C(R 00 )2, O, and S;

[0157] Q2is each occurrence, independently selected from the group consisting of a single bond, NR 00 , C(R00 )2, O, and S;

[0158] R5is selected from the group consisting of H, halogen, C(=O)R0, C(=O)OR0, substituted or unsubstituted C1-C6alkyl, C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl; in an embodiment, R5is selected from the group consisting of H, halogen, C(=O)R0, C(=O)OR0, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, and substituted or unsubstituted C2-C6alkynyl, the C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, and C2-C6alkynyl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C6alkyl;

[0159] Q4is CR0or N, and Q3is CR0or N;

[0160] Q5is NR0;

[0161] X2, X3, X4, X5, and X6are each independently selected at each occurrence from the group consisting of H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, C1-C6alkoxy, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; in an embodiment,

[0162] X2, X3, X4, X5, and X6are each independently selected at each occurrence from the group consisting of H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, C1-C6alkoxy, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, the C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C6alkyl;

[0163] R0and R 00each occurrence is independently selected from the group consisting of H, OH, N02, NH2, SH, CN, halogen, -C(=0)H, -C(=0)-Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; in an embodiment, R0and R 00 each occurrence is independently selected from the group consisting of H, OH, N02, NH2, SH, CN, halogen, -C(=0)H, -C(=0)-Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; in an embodiment, R0and R 1-6 substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said Ci-C6alkyl, Ci-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and Ci-C6alkyl; in an embodiment, R0and R 00 each occurrence is independently selected from the group consisting of H, OH, N02, NH2, SH, CN, halogen, -C(=0)H, -C(=0)-Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; in an embodiment, R0and R 1-3 substituted or unsubstituted Ci-C3alkyl, substituted or unsubstituted Ci-C3alkoxy, substituted or unsubstituted C2-C3alkenyl, substituted or unsubstituted C2-C3alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said Ci-C3alkyl, Ci-C3alkoxy, C2-C3alkenyl, C2-C3alkynyl, aryl, and heteroaryl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and Ci-C3alkyl; in an embodiment, R0and R 00 each occurrence is independently selected from the group consisting of H, OH, F, Cl, Br, I, substituted or unsubstituted Ci-C3alkoxy, and substituted or unsubstituted Ci-C3alkyl, said Ci-C3alkyl, and Ci-C3alkoxy being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and Ci-C3alkyl;

[0164] n1 is selected from 0, 1, 2, 3, 4;

[0165] La is selected from a single bond, -0-, C2-C6alkenylene, C2-C6alkynylene, Ci-C6alkylene, Ci-C6haloalkylene, -NR 00- C(O)-, -C(S)-, -SO2-, C3-C6cycloalkylene, C3-C6heterocycloalkylene containing 1, 2, or 3 heteroatoms independently selected from N, O, and S; in an embodiment, La is selected from the group consisting of a single bond, -O-, C2-C3alkynylene, C1-C3alkylene, C1-C3haloalkylene, -NH-, -NCH3-, -C(O)-, -C(S)-, -SO2-, C3-C6cycloalkylene, and C3-C6heterocycloalkylene containing 1, 2, or 3 heteroatoms independently selected from N, O, and S; in an embodiment, La is selected from the group consisting of a single bond, -NH-, -(CH2) m3 O-, -(CH2) m3 N(CH3)-, -(CH2) m3 NH-,

[0166] each occurrence of m1is independently selected from 0, 1, and 2; in an embodiment, each occurrence of m1is independently 1;

[0167] each occurrence of m2is independently selected from 1 and 2; in an embodiment, each occurrence of m2is independently 1;

[0168] m3is selected from 0, 1, 2, 3, 4, and 5;

[0169] m4is selected from 0, 1, 2, 3, 4, and 5; in an embodiment, m4is selected from 0 and 1;

[0170] L0is L j -L p -L 2 -L 3 -L 4 -L 5 -;

[0171] wherein,

[0172] L j is wherein, X S is a single bond or N(C 1-6 alkyl); each R is independently a thiol-reactable group;

[0173] L pis selected from a single bond, 6-10 membered arylene, and 5-8 membered heteroarylene comprising 1 to 3 heteroatoms independently selected from N, O, and S as ring atoms, wherein each of said 6-10 membered arylene and 5-8 membered heteroarylene is independently optionally substituted with one or more R 1L substituents;

[0174] L 2 is a single bond, or a combination of one or more of alkynylene, alkenylene, NR 2L , O, C(O), alkylene, and heteroalkylene, wherein said alkynylene, alkenylene, alkylene, and heteroalkylene are optionally substituted with one or more R 2L substituents;

[0175] L 3 is wherein, r1 is selected from a natural number between 1-20; r2 is selected from a natural number between 0-20; r3 is selected from 1, 2, 3; r4 is selected from 0, 1, 2, 3, 4, 5; r5 is selected from an integer between 1-30, wherein, L2 represents the connection site to L 2 , and L4 represents the connection site to L 4 ; in an embodiment thereof, r1 is selected from 1, 2, 3, 4, 5, 6; r2 is selected from 0, 1, 2, 3, 4, 5, 6; r4 is selected from 0, 1, 2; r5 is selected from an integer between 5-15, in an embodiment thereof, r5 is selected from 10, 11, 12, 13, 14, 15,

[0176] Ar 1 is, at each occurrence, independently selected from triazolylene, phenylene, pyrrolidinylene, piperidinylene, cyclohexanylene, thiazolylene, and dihydrodihydropyridazinylene, said triazolylene, phenylene, pyrrolidinylene, piperidinylene, cyclohexanylene, thiazolylene, and dihydrodihydropyridazinylene being optionally substituted with one or more substituents selected from hydroxy, cyano, amino, C 1-6 alkyl, haloC 1-6 alkyl, C 1-6 alkoxy, haloC 1-6 alkoxy, and C 3-8 cycloalkyl;

[0177] in an embodiment thereof, Ar 1 is, at each occurrence, independently selected from triazolylene, phenylene, pyrrolidinylene, piperidinylene, cyclohexanylene, thiazolylene, and dihydrodihydropyridazinylene, said triazolylene, phenylene, pyrrolidinylene, piperidinylene, cyclohexanylene, thiazolylene, and dihydrodihydropyridazinylene being optionally substituted with one or more substituents selected from hydroxy, cyano, amino, and C 1-6 alkyl;

[0178] In one embodiment, Ar 1 each occurrence is selected from In one embodiment, Ar 1 each occurrence is independently

[0179] L 4 is a peptide containing 2 to 7 amino acid residues, wherein the amino acids are optionally substituted with one or more substituents selected from halogen, hydroxyl, cyano, amino, alkyl, haloalkyl, deuterium- substituted alkyl, alkoxy, haloalkoxy, and cycloalkyl;

[0180] L 5 is selected from a single bond, L4 -NR 2L (CR 3L R 4L ) t -Z1-(CR 3L R 4L ) t -C(=O)-* D0 , L4 -NR 2L (CR 3L R 4L ) t - D0 , L4 -NR 2L (CR 3L R 4L ) t -Z1-(CR 3L R 4L ) t -Z1-C(=O)-* D0 , L4 -NR 2L -(CR 3L R 4L ) t -Z1-Ar 3 -C(=O)-* D0 , L4 -NR 2L -Ar 2 -(CR 3L R 4L ) t -Z1-C(=O)-NR 2L -(CR 3L R 4L ) t -Ar 2 -NR 2L -C(=O)-* D0and L4 -NR 2L -Ar 2 -(CR 3L R 4L ) t -Z1-C(=O)-* D0 ;* L4 denotes the point of attachment to L 4 ,* D0 denotes the point of attachment to D0;

[0181] wherein t is independently at each occurrence selected from 0, 1, 2, 3, 4, 5, and 6; Z1is independently at each occurrence a single bond, O, S, or NH; Ar 2 is independently at each occurrence an arylene or heteroarylene group, the heteroarylene group containing 1, 2, or 3 heteroatoms independently selected from N, O, and S as ring atoms, wherein the arylene or heteroarylene group is optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxyl, cyano, C 1-6 alkyl, -O-C 1-6 alkyl, amino, -NH(C 1-6 alkyl), and -N(C 1-6 alkyl)2;

[0182] Ar 3 is independently at each occurrence a cycloalkylene or heterocycloalkylene group, the heterocycloalkylene group containing 1, 2, or 3 heteroatoms independently selected from N, O, and S as ring atoms, wherein the cycloalkylene or heterocycloalkylene group is optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxyl, cyano, C 1-6 alkyl, -O-C 1-6 alkyl, amino, -NH(C 1-6 alkyl), and -N(C 1-6 alkyl)2;

[0183] R 1L , R 2L , R 3L , R 4L , and R 5L are each independently at each occurrence selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 alkoxy, haloC 1-6 alkyl, haloC 1-6 alkyl, deuterated C 1-6 alkyl, carboxyl, -C 1-6 alkylene-NH2, and -C 1-6 alkylene-OH;

[0184] denotes the point of attachment.

[0185] In one embodiment, wherein:

[0186] R1is selected from H, F, Cl, Br, I, hydroxyl, -OC(=O)O-C1-C6alkyl, and C1-C6alkyl; in one embodiment, R1is selected from H and -OC(=O)O-C1-C3alkyl; in one embodiment, R1is -OC(=O)CH3; and / or

[0187] R2is selected from H, F, Cl, Br, I, hydroxyl, and C1-C6alkyl; in one embodiment, R2is selected from H, F, Cl, Br, I, hydroxyl, and C1-C3alkyl; in one embodiment, R2is methyl; and / or

[0188] Y1, Y2, Y3, Y4, Y5, Y6, Y8, and Y9are each independently selected from H, C1-C3alkyl, C1-C3alkoxy, C1-C3haloalkyl, OH, NO2, NH2, SH, CN, or halogen; in one embodiment, Y1is OH; and / or

[0189] Y2is -OCH3; and / or

[0190] Y3is CH3; and / or

[0191] Y4, Y5, Y6, Y8, and Y9are each independently H; and / or

[0192] Y7is selected from H, C1-C6alkyl, and C1-C6alkoxy; in one embodiment, Y7is selected from H, C1-C3alkyl, and C1-C3alkoxy; in one embodiment, Y7is selected from H, and C1-C3alkyl; Y7is C1-C3alkyl; in one embodiment, Y7is CH3;

[0193] X1is each occurrence independently selected from halogen, C1-C6alkyl, C1-C6haloalkyl, hydroxyl, C1-C6alkoxy, CN, (=O), and H; in one embodiment, X1is each occurrence independently selected from F, Cl, Br, I, C1-C3alkyl, C1-C3haloalkyl, hydroxyl, C1-C3alkoxy, CN, and H; in one embodiment, X1is selected from hydroxyl and CN;

[0194] Q1is selected from O, S, C(C1-C6alkyl)2, C(C1-C6alkyl)H, CH2, N(C1-C6alkyl), and NH; in one embodiment, Q1is selected from O, S, C(C1-C3alkyl)2, C(C1-C3alkyl)H, CH2, N(C1-C3alkyl), and NH; in one embodiment, Q1is selected from O, CH2, and NH; in one embodiment, Q1is selected from O and NH; and / or

[0195] Q2is selected from the group consisting of O, S, C(Ci-C6alkyl)2, C(Ci-C6alkyl)H, CH2, N(Ci-C6alkyl), and NH; in an embodiment, Q2is selected from the group consisting of O, S, C(Ci-C3alkyl)2, C(Ci-C3alkyl)H, CH2, N(Ci-C3alkyl), and NH; in an embodiment, Q2is selected from the group consisting of O, CH2, and NH; and / or

[0196] Q4is CH or N, Q3is CH or N, at least one of Q3and Q4is N; and / or

[0197] Q5is NH; and / or

[0198] R5is selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and Ci-C6alkyl; in an embodiment, R5is selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and Ci-C3alkyl; in an embodiment, R5is H; and / or

[0199] R3and R4are each independently selected from the group consisting of CH, C-F, C-Cl, C-Br, C-I, C-hydroxyl, and C-Ci-C3alkyl; in an embodiment, R3and R4are each independently CH; and R3and R4, together with the atoms to which they are attached, form formula (II-A), formula (II-B), formula (II-C), or (II-F), in an embodiment, R3and R4, together with the atoms to which they are attached, form in an embodiment, R3and R4, together with the atoms to which they are attached, form in an embodiment, R3and R4, together with the atoms to which they are attached, form

[0200] X2, X3, X4, and X5are each independently selected at each occurrence from the group consisting of H, F, Cl, Br, I, hydroxyl, and Ci-C6alkyl; in an embodiment, X2, X3, X4, and X5are each independently selected at each occurrence from the group consisting of H, F, Cl, Br, I, hydroxyl, and Ci-C3alkyl; in an embodiment, X2, X3, X4, and X5are each independently H at each occurrence; and / or

[0201] X6is selected from the group consisting of H, F, CI, Br, I, hydroxyl, C1-C6alkyl, -C1-C6alkyl-OH, and -C1-C6alkyl-NH2; in an embodiment, X6is selected from the group consisting of H, F, CI, Br, I, hydroxyl, C1-C3alkyl, -C1-C3alkyl-OH, and -C1-C3alkyl-NH2; X6is selected from the group consisting of H, -CH2-OH, and -CH2-NH2; and / or

[0202] R0is independently at each occurrence selected from the group consisting of H and C1-C3alkyl; in a further embodiment, R0is independently at each occurrence selected from the group consisting of H and CH3; and / or

[0203] R 00 is independently at each occurrence selected from the group consisting of H, F, CI, Br, I, hydroxyl, and C1-C6alkyl; in an embodiment, R 00 is independently at each occurrence selected from the group consisting of H, F, CI, Br, I, hydroxyl, and C1-C3alkyl; and / or

[0204] n1is selected from the group consisting of 0, and 1 ; and / or

[0205] in an embodiment, La is selected from the group consisting of a single bond,

[0206] in an embodiment, wherein:

[0207] L j is a single bond; wherein X S is selected from the group consisting of a single bond, NH and N-CH3; in an embodiment, X S is a single bond;

[0208] in an embodiment, L j is selected from the group consisting of

[0209] each R is independently selected from the group consisting of halogen and -S-Ar 4 ; wherein Ar 4 is selected from the group consisting of phenyl, C 1-6 alkylphenyl-, C 1-6 alkoxyphenyl-, 2-pyridyl, 2-pyrimidinyl, 1 -methylimidazol-2-yl, the phenyl, C 1-6 alkylphenyl-, C 1-6 alkoxyphenyl-, 2-pyridyl, 2-pyrimidinyl and 1 -methylimidazol-2-yl are optionally substituted with 1, 2, 3 or 4 R a ;

[0210] wherein W2is selected from the group consisting of amino, -NRa -C 1-6 alkyl, -NR a -C 1-6 alkoxy, -NR a -C 1-6 alkylene-NR b R c , -NR a -C 1-6 alkylene-C 1-6 alkoxy and 5- to 8-membered heterocycloalkyl containing 1, 2, 3 or 4 heteroatoms as ring members, selected from the group consisting of N, O and S, wherein the S atom is optionally substituted with 1 or 2 oxo groups; in one embodiment, W2 is selected from the group consisting of amino, -NR a -C 1-6 alkyl,

[0211] R a each occurrence is independently selected from the group consisting of halogen, C 1-6 alkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, deuterated C 1-6 alkyl, carboxyl, -C 1-6 alkylene-C 1-6 alkoxy, -C 1-6 alkylene-NH2 and -C 1-6 alkylene-OH;

[0212] R b , R c each occurrence is independently selected from the group consisting of C 1-6 alkyl and C 1-6 alkoxy;

[0213] In one embodiment, R each occurrence is independently selected from the group consisting of halogen, More in one embodiment, each R is independently selected from the group consisting of chlorine, bromine,

[0214] In one embodiment, L j is selected from the group consisting of

[0215] In one embodiment, wherein

[0216] L p is selected from the group consisting of a single bond, phenylene and 5-8 membered heteroarylene; said 5-8 membered heteroarylene comprises 1 to 3 heteroatoms independently selected from N, O and S as ring members; wherein said phenylene and 5-8 membered heteroarylene are each independently optionally substituted with one or more R 5L ;

[0217] In one embodiment thereof, each of said phenylene and 5-8 membered heteroarylene is independently optionally substituted with 1, 2, 3, 4, 5 or 6 R 5L substituents;

[0218] In one embodiment thereof, L p is selected from a single bond, phenylene and pyridinylene; wherein each of said phenylene and pyridinylene is independently optionally substituted with 1, 2, 3 or 4 R 5L substituents; and / or

[0219] R 5L is, at each occurrence, independently selected from halogen, hydroxyl, cyano, amino, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, haloC 1-6 alkoxy and C 3-8 cycloalkyl;

[0220] In one embodiment thereof, R 5L is, at each occurrence, independently selected from halogen, hydroxyl, cyano, C 1-3 alkyl and -O-C 1-3 alkyl;

[0221] In one embodiment thereof, R 5L is, at each occurrence, independently selected from Cl, Br, F, -OH, -CN, methyl and -OCH 3； In one embodiment thereof, R 5L is F;

[0222] In one embodiment thereof, L p is selected from a single bond, phenylene and pyridinylene; wherein each of said phenylene and pyridinylene is independently optionally substituted with 1, 2, 3 or 4 substituents selected from Cl, Br, F, -OH, -CN, methyl and -OCH3;

[0223] In one embodiment thereof, L p is selected from wherein Lj denotes the site of attachment to L j denotes the site of attachment to L L2 denotes the site of attachment to L 2 denotes the site of attachment to L

[0224] In one embodiment thereof, wherein

[0225] L 4 is a peptide comprising 2, 3, 4, 5 or 6 amino acids, wherein said amino acids are optionally substituted with one or more substituents selected from halogen, hydroxyl, cyano, amino, C 1-6 alkyl, haloC 1-6alkyl, deuterated C 1-6 alkyl, C 1-6 alkoxy and C 5-8 cycloalkyl; in an embodiment thereof, L 4 is a peptide comprising 2 to 6 amino acid residues selected from the group consisting of glycine, phenylalanine, alanine, valine, lysine, citrulline, serine, glutamic acid and aspartic acid;

[0226] in an embodiment thereof, L 4 is a dipeptide, tripeptide or tetrapeptide composed of amino acid residues selected from the group consisting of alanine, phenylalanine, valine, glycine, lysine and citrulline;

[0227] in an embodiment thereof, L 4 is a peptide selected from the group consisting of glycine-phenylalanine-glycine, alanine-alanine-alanine-glycine, alanine-alanine-alanine, glycine-glycine-phenylalanine, glycine-glycine-phenylalanine-glycine, valine-citrulline and valine-alanine;

[0228] wherein the peptide is optionally substituted by one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, amino, alkyl, haloalkyl, deuterated alkyl, alkoxy and cycloalkyl, in an embodiment thereof, is optionally substituted by one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, amino and C 1-6 alkyl substituents;

[0229] in an embodiment thereof, L 4 is selected from wherein L3 denotes the site of attachment to L 3 denotes the site of attachment to L L5 denotes the site of attachment to L 5 denotes the site of attachment to L

[0230] in an embodiment thereof,

[0231] wherein t is at each occurrence independently from another 0, 1 or 2; and / or

[0232] Z1is at each occurrence independently from another a single bond, O or S; preferably a single bond or O; more preferably O; and / or

[0233] Ar 2 is at each occurrence independently from another phenylene, which is optionally substituted by one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, C 1-6 alkyl, -O-C 1-6 alkyl, amino, -NH(C 1-6 alkyl) and -N(C 1-6substituted with one or more substituents selected from halo, hydroxyl, cyano, C 2 independently at each occurrence is

[0234] Ar 3 independently at each occurrence is 4-6 membered cycloalkylene, optionally substituted with one or more substituents selected from halo, hydroxyl, cyano, C 1-6 alkyl, -O-C 1-6 alkyl, amino, -NH(C 1-6 alkyl) and -N(C 1-6 alkyl)2; in an embodiment thereof, Ar 3 independently at each occurrence is

[0235] in an embodiment thereof, L 5 is selected from a single bond, and / or

[0236] L 2 is O; and / or

[0237] L 3 is selected from

[0238] in an embodiment thereof, the L0is selected from:

[0239] represents the point of attachment of L0to D0.

[0240] in an embodiment thereof, wherein D0is selected from represents the point of attachment of D0to L0.

[0241] in an embodiment thereof, is selected from

[0242] A fifth aspect of the present application provides an antibody drug conjugate of Formula III, or a pharmaceutically acceptable salt thereof,p (Formula III)

[0243] wherein Ab is an antibody or antigen binding fragment thereof;

[0244] p is an integer or decimal number from 0 to 20, but not 0;

[0245] L is a linker that is -T-L p -L 2 -L 3 -L 4 -L 5 -;

[0246] L p , L 2 , L 3 , L 4 , L 5 , and D0 are as defined in any of the preceding;

[0247] T is selected from

[0248] In one embodiment, D0 is as shown in Formula I-2,

[0249] wherein R1 and R2 are each independently selected from H, halogen, hydroxyl, C(=O)R0, C(=O)OR0, -OC(=O)OR0, -OC(=O)R0, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, and substituted or unsubstituted C1-C6 haloalkyl; in one embodiment, R1 and R2 are each independently selected from H, halogen, hydroxyl, C(=O)R0, C(=O)OR0, -OC(=O)OR0, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, and substituted or unsubstituted C1-C6 haloalkyl, said C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, and C1-C6 haloalkyl optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6 alkyl;

[0250] Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, and Y9 are each independently selected from H, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, OH, NO2, NH2, SH, CN, or halogen;

[0251] X1is each occurrence independently selected from halogen, alkyl, haloalkyl, OR x, CN, (=0) and H; wherein R x is selected from H, alkyl, haloalkyl, alkenyl, alkynyl, OH, OR0, SR0, SOR0, and SO2R0;

[0252] R3and R4are each independently selected from CH, C-F, C-Cl, C-Br, C-I, C-hydroxyl, and C-C1-C6alkyl; and R3and R4, together with the atoms to which they are attached, form a

[0253] Q1is, at each occurrence, independently selected from NR 00 , C(R 00 )2, O, and S;

[0254] Q2is, at each occurrence, independently selected from a single bond, NR 00 , C(R 00 )2, O, and S;

[0255] R5is selected from H, halogen, C(=O)R0, C(=O)OR0, substituted or unsubstituted C1-C6alkyl, C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl; in an embodiment, R5is selected from H, halogen, C(=O)R0, C(=O)OR0, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, and substituted or unsubstituted C2-C6alkynyl, the C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, and C2-C6alkynyl being optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6alkyl;

[0256] Q4is CR0or N, Q3is CR0or N;

[0257] Q5is NR0;

[0258] X2, X3, X4, X5, and X6are each independently at each occurrence selected from H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, C1-C6alkoxy, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; in an embodiment,

[0259] X2, X3, X4, X5, and X6are each independently selected, at each occurrence, from H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6alkyl;

[0260] R0and R 00 are each independently selected, at each occurrence, from H, OH, NO2, NH2, SH, CN, halogen, -C(=O)H, -C(=O)-C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; in one embodiment, R0and R 00 are each independently selected, at each occurrence, from H, OH, NO2, NH2, SH, CN, F, Cl, Br, I, -C(=O)H, -C(=O)-C 1-6 alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; in one embodiment, R0and R 00 are each independently selected, at each occurrence, from H, OH, NO2, NH2, SH, CN, F, Cl, Br, I, -C(=O)H, -C(=O)-C 1-3 alkyl, substituted or unsubstituted C1-C3alkoxy, substituted or unsubstituted C1-C3alkyl, substituted or unsubstituted C2-C3alkenyl, substituted or unsubstituted C2-C3alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said C1-C3alkyl, C1-C3alkoxy, C2-C3alkenyl, C2-C3alkynyl, aryl, and heteroaryl optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C3alkyl; in one embodiment, R0and R 00each occurrence is selected independently from H, OH, F, CI, Br, I, substituted or unsubstituted C1-C3alkoxy, and substituted or unsubstituted C1-C3alkyl, said C1-C3alkyl, and C1-C3alkoxy optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C3alkyl;

[0261] n1 is selected from 0, 1, 2, 3, 4;

[0262] La is selected from a single bond, -O-, C2-C6alkenylene, C2-C6alkynylene, C1-C6alkylene, C1-C6haloalkylene, -NR 00 -, -C(O)-, -C(S)-, -SO2-, C3-C6cycloalkylene, C3-C6heterocycloalkylene containing 1, 2, or 3 heteroatoms independently selected from N, O, and S; in an embodiment, La is selected from a single bond, -O-, C2-C3alkynylene, C1-C3alkylene, C1-C3haloalkylene, -NH-, -NCH3-, -C(O)-, -C(S)-, -SO2-, C3-C6cycloalkylene, and C3-C6heterocycloalkylene containing 1, 2, or 3 heteroatoms independently selected from N, O, and S; in an embodiment, La is selected from a single bond, -NH-, -(CH2) m3 O-, -(CH2) m3 N(CH3)-, -(CH2) m3 NH-,

[0263] each occurrence of m1 is independently selected from 0, 1, and 2; in an embodiment, each occurrence of m1 is independently 1;

[0264] each occurrence of m2 is independently selected from 1 and 2; in an embodiment, each occurrence of m2 is independently 1;

[0265] m3 is selected from 0, 1, 2, 3, 4, and 5;

[0266] m4 is selected from 0, 1, 2, 3, 4, and 5; in an embodiment, m4 is selected from 0 and 1;

[0267] L p is selected from a single bond, 6-10 membered aralkylene, and 5-8 membered heteroaralkylene comprising 1 to 3 heteroatoms independently selected from N, O, and S as ring atoms, wherein each of said 6-10 membered aralkylene, and 5-8 membered heteroaralkylene is independently optionally substituted with one or more R1L substituted;

[0268] L 2 is a single bond, or a combination of one or more of alkynylene, alkenylene, NR 2L , O, C(O), alkylene, and heteroalkylene, wherein the alkynylene, alkenylene, alkylene, and heteroalkylene are optionally substituted with one or more R 2L substituted;

[0269] L 3 is wherein, r1 is selected from a natural number between 1-20; r2 is selected from a natural number between 0-20; r3 is selected from 1, 2, 3; r4 is selected from 0, 1, 2, 3, 4, 5; r5 is selected from an integer between 1-30, wherein, L2 represents the connection site with L 2 , and L4 represents the connection site with L 4 ; in an embodiment thereof, r1 is selected from 1, 2, 3, 4, 5, 6; r2 is selected from 0, 1, 2, 3, 4, 5, 6; r4 is selected from 0, 1, 2; r5 is selected from an integer between 5-15, in an embodiment thereof, r5 is selected from 10, 11, 12, 13, 14, 15,

[0270] Ar 1 is independently selected at each occurrence from triazolylene, phenylene, pyrrolidinylene, piperidinylene, cyclohexanylene, thiazolylene, and dihydrodihydropyridazinylene, optionally substituted with one or more substituents selected from hydroxy, cyano, amino, C 1-6 alkyl, haloC 1-6 alkyl, C 1-6 alkoxy, haloC 1-6 alkoxy, and C 3-8 cycloalkyl;

[0271] in an embodiment thereof, Ar 1 is independently selected at each occurrence from triazolylene, phenylene, pyrrolidinylene, piperidinylene, cyclohexanylene, thiazolylene, and dihydrodihydropyridazinylene, optionally substituted with one or more substituents selected from hydroxy, cyano, amino, and C 1-6 alkyl;

[0272] in an embodiment thereof, Ar 1 is independently selected at each occurrence from in an embodiment thereof, Ar 1Each time it appears, it is independent.

[0273] L 4 It is a peptide containing 2 to 7 amino acid residues, wherein the amino acid is optionally substituted by one or more substituents selected from halogen, hydroxyl, cyano, amino, alkyl, haloalkyl, deuteryl, alkoxy, haloalkoxy and cycloalkyl.

[0274] L 5 Selected from single key, * L4 -NR 2L (CR 3L R 4L ) t -Z1-(CR 3L R 4L ) t -C(=O)-* D0 、* L4 -NR 2L (CR 3L R 4L ) t -* D0 、* L4 -NR 2L (CR 3L R 4L ) t -Z1-(CR 3L R 4L ) t -Z1-C(=O)-* D0 、* L4 -NR 2L -(CR 3L R 4L ) t -Z1-Ar 3 -C(=O)-* D0 、* L4 -NR 2L -Ar 2 -(CR 3L R 4L ) t -Z1-C(=O)-NR 2L -(CR 3L R 4L ) t -Ar 2 -NR 2L -C(=O)-* D0 and* L4 -NR 2L -Ar 2 -(CR 3L R 4L ) t-Z1-C(=0)-* D0 L4 represents the point of attachment to L 4 D0 represents the point of attachment to D0;

[0275] wherein t is independently at each occurrence selected from 0, 1, 2, 3, 4, 5, and 6; Z1is independently at each occurrence a single bond, O, S, or NH; Ar 2 is independently at each occurrence arylene or heteroarylene, the heteroarylene containing 1, 2, or 3 heteroatoms independently selected from N, O, and S as ring atoms, wherein the arylene or heteroarylene is optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxyl, cyano, C 1-6 alkyl, -O-C 1-6 alkyl, amino, -NH(C 1-6 alkyl), and -N(C 1-6 alkyl)2;

[0276] Ar 3 is independently at each occurrence cycloalkylene or heterocycloalkylene, the heterocycloalkylene containing 1, 2, or 3 heteroatoms independently selected from N, O, and S as ring atoms, wherein the cycloalkylene or heterocycloalkylene is optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxyl, cyano, C 1-6 alkyl, -O-C 1-6 alkyl, amino, -NH(C 1-6 alkyl), and -N(C 1-6 alkyl)2;

[0277] R 1L , R 2L , R 3L , R 4L , and R 5L are each independently at each occurrence selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 alkoxy, haloC 1-6 alkyl, haloC 1-6 alkyl, deuterated C 1-6 alkyl, carboxyl, -C 1-6 alkylene-NH2, and -C 1-6 alkylene-OH;

[0278] T is selected from

[0279] represents the point of attachment.

[0280] In an embodiment, wherein the linker is selected from: ​​​

[0281] wherein, represents the point of attachment of the linker to D0. In one embodiment, it is selected from:

[0282] p is selected from an integer or decimal number from 1 to 12; in one embodiment, p is selected from an integer or decimal number from 1 to 10; in one embodiment, p is selected from an integer or decimal number from 2 to 8; in one embodiment, p is selected from an integer or decimal number from 2 to 6; preferably p is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10.

[0283] In one embodiment, wherein the antibody is selected from a chimeric antibody, a humanized antibody and a fully human antibody;

[0284] In one embodiment, the antibody or antigen binding fragment thereof is selected from an anti-TROP-2 antibody, an anti-HER2 (ErbB2) antibody, an anti-NECTIN4 antibody, an anti-EGFR antibody, an anti-B7-H3 antibody, an anti-c-Met antibody, an anti-HER3 (ErbB3) antibody, an anti-HER4 (ErbB4) antibody, an anti-LIV-1 antibody, an anti-ROR1 antibody, an anti-CD20 antibody, an anti-CD22 antibody, an anti-CD30 antibody, an anti-CD33 antibody, an anti-CD44 antibody, an anti-CD56 antibody, an anti-CD70 antibody, an anti-CD73 antibody, an anti-CD105 antibody, an anti-CEA antibody, an anti-A33 antibody, an anti-Cripto antibody, an anti-EphA2 antibody, an anti-G250 antibody, an anti-MUC1 antibody, an anti-Lewis Y antibody, an anti-VEGFR antibody, an anti-GPNMB antibody, an anti-Integrin antibody, an anti-PSMA antibody, an anti-Tenascin-C antibody, an anti-SLC44A4 antibody, an anti-Mesothelin antibody, an anti-CDH17 antibody and an anti-GPC3 antibody or an antigen binding fragment thereof;

[0285] In one embodiment, 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, an anti-CDH17 antibody and an anti-GPC3 antibody or an antigen binding fragment thereof;

[0286] In one embodiment, the antibody or antigen-binding fragment thereof is an anti-HER2 (ErbB2) antibody, an anti-NECTIN4 antibody, an anti-B7-H3 antibody, an anti-CDH17 antibody, and an anti-GPC3 antibody or antigen-binding fragment thereof

[0287] In one embodiment, the antibody or antigen-binding fragment thereof is an anti-NECTIN4 antibody or antigen-binding fragment thereof; in one embodiment, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the heavy chain comprises a heavy chain variable region comprising 3 heavy chain complementarity determining regions (HCDRs), wherein the amino acid sequence of HCDR1 is set forth in SEQ ID NO: 11, the amino acid sequence of HCDR2 is set forth in SEQ ID NO: 12, and the amino acid sequence of HCDR3 is set forth in SEQ ID NO: 13; and / or the light chain comprises a light chain variable region comprising 3 light chain complementarity determining regions (LCDRs), wherein the amino acid sequence of LCDR1 is set forth in SEQ ID NO: 14, the amino acid sequence of LCDR2 is set forth in SEQ ID NO: 15, and the amino acid sequence of LCDR3 is set forth in SEQ ID NO: 16; in one embodiment, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and / or a light chain variable region, wherein the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 18; and / or the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 17; in one embodiment, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is set forth in SEQ ID NO: 1, and / or the amino acid sequence of the light chain is set forth in SEQ ID NO: 2;

[0288] In an embodiment, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the heavy chain comprises a heavy chain variable region comprising 3 heavy chain complementarity determining regions (HCDRs), wherein the amino acid sequence of HCDR1 is set forth in SEQ ID NO: 19, the amino acid sequence of HCDR2 is set forth in SEQ ID NO: 20, and the amino acid sequence of HCDR3 is set forth in SEQ ID NO: 21; and / or the light chain comprises a light chain variable region comprising 3 light chain complementarity determining regions (LCDRs), wherein the amino acid sequence of LCDR1 is set forth in SEQ ID NO: 22, the amino acid sequence of LCDR2 is set forth in SEQ ID NO: 23, and the amino acid sequence of LCDR3 is set forth in SEQ ID NO: 24; in an embodiment, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and / or a light chain variable region, wherein the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 26; and / or the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 25; in an embodiment, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is set forth in SEQ ID NO: 3, and / or the amino acid sequence of the light chain is set forth in SEQ ID NO: 4;

[0289] In an embodiment, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the heavy chain comprises a heavy chain variable region comprising 3 heavy chain complementarity determining regions (HCDRs), wherein the amino acid sequence of HCDR1 is set forth in SEQ ID NO: 27, the amino acid sequence of HCDR2 is set forth in SEQ ID NO: 28, and the amino acid sequence of HCDR3 is set forth in SEQ ID NO: 29; and / or the light chain comprises a light chain variable region comprising 3 light chain complementarity determining regions (LCDRs), wherein the amino acid sequence of LCDR1 is set forth in SEQ ID NO: 30, the amino acid sequence of LCDR2 is set forth in SEQ ID NO: 31, and the amino acid sequence of LCDR3 is set forth in SEQ ID NO: 32; in an embodiment, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and / or a light chain variable region, wherein the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 34; and / or the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 33; in an embodiment, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is set forth in SEQ ID NO: 5, and / or the amino acid sequence of the light chain is set forth in SEQ ID NO: 6;

[0290] In an embodiment, the antibody or antigen-binding fragment thereof is an anti-B7-H3 antibody or antigen-binding fragment thereof; in an embodiment, the anti-B7-H3 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the heavy chain comprises a heavy chain variable region comprising three heavy chain complementarity determining regions (HCDRs), wherein the amino acid sequence of HCDR1 is set forth in SEQ ID NO: 35, the amino acid sequence of HCDR2 is set forth in SEQ ID NO: 36, and the amino acid sequence of HCDR3 is set forth in SEQ ID NO: 37; and / or the light chain comprises a light chain variable region comprising three light chain complementarity determining regions (LCDRs), wherein the amino acid sequence of LCDR1 is set forth in SEQ ID NO: 38, the amino acid sequence of LCDR2 is set forth in SEQ ID NO: 39, and the amino acid sequence of LCDR3 is set forth in SEQ ID NO: 40; in an embodiment, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and / or a light chain variable region, wherein the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 42; and / or the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 41; in an embodiment, the anti-B7-H3 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is set forth in SEQ ID NO: 7, and / or the amino acid sequence of the light chain is set forth in SEQ ID NO: 8;

[0291] In one embodiment, the anti-B7-H3 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the heavy chain comprises a heavy chain variable region comprising 3 heavy chain complementarity determining regions (HCDRs), wherein the amino acid sequence of HCDR1 is set forth in SEQ ID NO: 43, the amino acid sequence of HCDR2 is set forth in SEQ ID NO: 44, and the amino acid sequence of HCDR3 is set forth in SEQ ID NO: 45; and / or the light chain comprises a light chain variable region comprising 3 light chain complementarity determining regions (LCDRs), wherein the amino acid sequence of LCDR1 is set forth in SEQ ID NO: 46, the amino acid sequence of LCDR2 is set forth in SEQ ID NO: 47, and the amino acid sequence of LCDR3 is set forth in SEQ ID NO: 48; in one embodiment, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and / or a light chain variable region, wherein the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 50; and / or the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 49; in one embodiment, the anti-B7-H3 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is set forth in SEQ ID NO: 9, and / or the amino acid sequence of the light chain is set forth in SEQ ID NO: 10.

[0292] The seventh aspect of the present application provides a pharmaceutical composition consisting of substance X and a pharmaceutically acceptable excipient.

[0293] The substance X is substance X1 or substance X2; the substance X1 is a compound or a pharmaceutically acceptable salt thereof as described in any of the preceding; the substance X2 is an antibody-drug conjugate as described in any of the preceding and represented by formula III.

[0294] The eighth aspect of the present application provides use of a compound as described in any of the preceding, a drug linker conjugate as described in the preceding, an antibody-drug conjugate as described in any of the preceding, or a pharmaceutical composition as described in the preceding, in the manufacture of a medicament for treating or preventing a disease associated with abnormal cell activity; in one embodiment, the disease is cancer.

[0295] In one embodiment, the cancer is a cancer associated with abnormal expression of B7-H3, abnormal expression of HER2, abnormal expression of NECTIN4, abnormal expression of CDH17, or abnormal expression of GPC3.

[0296] In one embodiment, the cancer is selected from breast cancer, gastric cancer, pancreatic cancer, prostate cancer, colorectal cancer, melanoma, and lung cancer; in one embodiment, the prostate cancer includes androgen-dependent or androgen-independent prostate cancer; the lung cancer includes non-small cell lung cancer and small cell lung cancer; the colorectal cancer includes colorectal adenocarcinoma; the pancreatic cancer includes pancreatic ductal adenocarcinoma.

[0297] In one embodiment, the cancer is selected from breast cancer, gastric cancer, pancreatic cancer, prostate cancer, colorectal cancer, melanoma, and lung cancer; in one embodiment, the prostate cancer includes androgen-dependent or androgen-independent prostate cancer; the lung cancer includes non-small cell lung cancer and small cell lung cancer; the colorectal cancer includes colorectal adenocarcinoma; the pancreatic cancer includes pancreatic ductal adenocarcinoma.

[0298] In one embodiment, the cancer is selected from breast cancer, gastric cancer, pancreatic cancer, prostate cancer, colorectal cancer, melanoma, and lung cancer; in one embodiment, the prostate cancer includes androgen-dependent or androgen-independent prostate cancer; the lung cancer includes non-small cell lung cancer and small cell lung cancer; the colorectal cancer includes colorectal adenocarcinoma; the pancreatic cancer includes pancreatic ductal adenocarcinoma.

[0299] In one embodiment, the cancer is selected from breast cancer, gastric cancer, pancreatic cancer, prostate cancer, colorectal cancer, melanoma, and lung cancer; in one embodiment, the prostate cancer includes androgen-dependent or androgen-independent prostate cancer; the lung cancer includes non-small cell lung cancer and small cell lung cancer; the colorectal cancer includes colorectal adenocarcinoma; the pancreatic cancer includes pancreatic ductal adenocarcinoma.

[0300] In one embodiment, the cancer is selected from breast cancer, gastric cancer, pancreatic cancer, prostate cancer, colorectal cancer, melanoma, and lung cancer; in one embodiment, the prostate cancer includes androgen-dependent or androgen-independent prostate cancer; the lung cancer includes non-small cell lung cancer and small cell lung cancer; the colorectal cancer includes colorectal adenocarcinoma; the pancreatic cancer includes pancreatic ductal adenocarcinoma.

[0301] In one embodiment, the cancer is selected from breast cancer, gastric cancer, pancreatic cancer, prostate cancer, colorectal cancer, melanoma, and lung cancer; in one embodiment, the prostate cancer includes androgen-dependent or androgen-independent prostate cancer; the lung cancer includes non-small cell lung cancer and small cell lung cancer; the colorectal cancer includes colorectal adenocarcinoma; the pancreatic cancer includes pancreatic ductal adenocarcinoma.

[0302] The present application also provides a use of a compound as described in any of the preceding embodiments as a small molecule toxin for the preparation of an antibody drug conjugate.

[0303] The present application also provides the use of a compound as described in any of the preceding for the manufacture of an antibody-drug conjugate comprising a covalently attached antibody or antigen binding fragment thereof and a linker-small molecule toxin moiety, wherein the linker-small molecule toxin moiety is a drug linker conjugate as described in any of the preceding.

[0304] Terms and definitions

[0305] Unless otherwise defined herein, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

[0306] Certain compounds of the present application can exist in free form or, where appropriate, in the form of a pharmaceutically acceptable derivative thereof. In the present application, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, prodrugs, stereoisomers (including but not limited to diastereiomers and enantiomers), tautomers, solvates, polymorphs, and isotopologues, of the compounds, which upon administration to a patient in need thereof, are capable of becoming, directly or indirectly, the compounds of the present application or a metabolite thereof. Accordingly, as used herein, reference to "a compound of the present application" is meant to also encompass all such potential derivative forms.

[0307] The term "pharmaceutically acceptable salt" refers to salts that retain the biological effectiveness and nontoxic of the free acids and bases of the particular compound. Examples of pharmaceutically acceptable salts include, but are not limited to: (1) acid addition salts, formed with inorganic acids such as hydrochloric acid, sulfuric acid, hydrobromic acid, nitric acid, phosphoric acid, and the like; or 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, hydroxyacetic acid, cinnamic acid, pyruvic acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, propyleneuric acid, mandelic acid, and the like; or (2) base addition salts, formed with alkali metals, such as sodium, potassium, and the like; and alkaline earth metals, such as calcium, magnesium, and the like; and organic bases such as ammonium hydroxide, choline hydroxide, diethanolamine, lysine, ethylenediamine, t-butylamine, t-octylamine, tris(hydroxymethyl)aminomethane, N-methylglucamine, triethanolamine, dehydroabietylamine, and the like. Other pharmaceutically acceptable salts are known to those skilled in the art.

[0308] Prodrugs of the compounds of the present application are included within the scope of the present application. In general, such prodrugs will be functional derivatives of the compounds which are readily convertible in vivo into the required compound. Thus, the term "administration of a compound", as used herein, encompasses the

[0309] The compounds of the present application can contain one or more asymmetric centers and can thus give rise to enantiomers and diastereomers. The present application includes all possible enantiomers and diastereomers of the compounds of the present application, as well as, their racemic mixtures, their substantially pure resolved individual enantiomers, all possible geometric isomers, and pharmaceutically acceptable salts thereof.

[0310] The compounds of the present application are not defined with respect to the stereochemistry at any position of the compounds. The present application includes all stereoisomers and pharmaceutically acceptable salts of the compounds. Also, mixtures of stereoisomers and separated individual stereoisomers are included in the present application. The products of the syntheses can be mixtures of stereoisomers, which can be separated into their individual stereoisomers using methods known to those of ordinary skill in the art.

[0311] Where the compounds of the present application contain one or more chiral centers, and where the compounds possess any level of optical activity, the present application includes each of the individual enantiomers of the instant compounds. The present application also includes every possible stereoisomer of the compounds of the present application. Where the compounds of the present application contain one or more double bonds, and where the compounds possess any level of geometric isomerism, the present application includes each of the individual geometric isomers and mixtures thereof.

[0312] Where the compounds of the present application exist in solvated forms, the present application includes each of the possible solvated forms that the compounds are capable of existing in. The type of solvent molecules which can be present in the solvated forms are not specifically limited as long as the solvent is pharmacologically acceptable. For example, water, ethanol, propanol, acetone, and the like can be used.

[0313] The present application also includes all pharmaceutically acceptable isotopically enriched compounds which are the same as the compounds of the present application except for the inclusion of one or more isotopic atoms having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number present in nature. Examples of isotopes suitable for inclusion in the compounds of the present application include, but are not limited to, isotopes of hydrogen, such as deuterium (2H) and tritium (3H); isotopes of carbon, such as 13C and 14C; isotopes of chlorine, such as 37CI; isotopes of iodine, such as 125I; isotopes of nitrogen, such as 13N and 15N; isotopes of oxygen, such as 17O and 18O; isotopes of phosphorus, such as 32P; and isotopes of sulfur, such as 34S.

[0314] The term "ligand-drug conjugate" refers to a ligand linked to a biologically active drug through a stable linker. In the present disclosure, the "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 through a stable linker.

[0315] The term "ligand" refers to a macromolecular compound capable of recognizing and binding to an antigen or receptor associated with a target cell. The role of the ligand is to present the drug to the target cell population to which the ligand binds. In the embodiments of the present application, the ligand is represented as Ab, and the ligand can form a linkage with the linker through a heteroatom on the ligand. The ligand is preferably an antibody or an antigen-binding fragment thereof or a polypeptide, and the antibody is selected from a chimeric antibody, a humanized antibody, a fully human antibody, or a murine antibody; and is preferably a monoclonal antibody.

[0316] The term "ligand binding moiety" refers to a group capable of binding to a ligand.

[0317] The term "drug linking moiety" refers to a group capable of binding to a drug.

[0318] The term "drug", "toxin", or "payload" refers to a cytotoxic drug, which is a chemical molecule capable of strongly damaging the normal growth of tumor cells in the tumor cells. And has a reactive functional group to prepare the antibody drug conjugate or drug linker conjugate compound described in the present application.

[0319] The term "linker unit" or "linking fragment" or "linking unit" or "linking moiety" or "linker" refers to a chemical structure fragment or bond connected to the ligand at one end and connected to the drug at the other end, which can also be connected to other linkers before being connected to the drug.

[0320] The three-letter and one-letter codes for amino acids used in the present disclosure are as described in J. Biol. Chem, 243, p3558 (1968).

[0321] The term "antibody" refers to an immunoglobulin, which is a four polypeptide chain structure connected by interchain disulfide bonds of two identical heavy chains and two identical light chains. The antigenicity of immunoglobulin heavy chain constant region is different due to different amino acid composition and arrangement order. Accordingly, immunoglobulin can be divided into five types, or called isotypes of immunoglobulin, i.e. IgM, IgD, IgG, IgA and IgE, and the corresponding heavy chains are μ chain, δ chain, γ chain, α chain and ε chain, respectively. The same type of Ig can be divided into different subtypes according to the difference of amino acid composition of hinge region and the number and position of heavy chain disulfide bonds, such as IgG can be divided into IgG1, IgG2, IgG3 and IgG4. The light chain is divided into κ chain or λ chain by different constant regions. Each type of Ig in the five types of Ig can have κ chain or λ chain. The antibody described in the present disclosure is preferably a specific antibody against a cell surface antigen on a target cell, and non-limiting examples are 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-MUC1 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; more preferably, the antibody or antigen-binding fragment thereof is an anti-B7-H3 antibody, an anti-NECTIN4 antibody, an anti-HER2 (ErbB2) antibody or an antigen-binding fragment thereof.

[0322] The antibody of the present application includes murine antibody, chimeric antibody, humanized antibody and fully human antibody, preferably humanized antibody and fully human antibody.

[0323] The term "murine antibody" in the present application is an antibody prepared from a mouse according to the knowledge and skill in the art. When prepared, the test subject is injected with a specific antigen, and then hybridoma expressing antibody with desired sequence or functional characteristics is isolated.

[0324] The term "chimeric antibody" is an antibody in which the variable region of a murine antibody is fused with the constant region of a human antibody, and can reduce the immune response induced by the murine antibody. To construct a chimeric antibody, a hybridoma secreting a murine-specific monoclonal antibody is first constructed, and then the variable region gene is cloned from the murine hybridoma cell. The constant region gene of a human antibody is cloned as needed, and the murine variable region gene and the human constant region gene are ligated to form a chimeric gene, which is inserted into an expression vector, and finally the chimeric antibody molecule is expressed in a eukaryotic system or a prokaryotic system.

[0325] The term "humanized antibody" is also referred to as a CDR-grafted antibody, and refers to an antibody in which the CDR sequence of a mouse is grafted into the variable region framework of a human antibody, i.e., an antibody produced in a different type of human germline antibody framework sequence. It can overcome the heterogeneity reaction induced by the chimeric antibody carrying a large amount of murine protein component.

[0326] The term "fully human antibody", "human antibody" or "completely human antibody", also referred to as "fully human monoclonal antibody", is an antibody in which the variable region and the constant region of the antibody are both human-derived, and which is free of immunogenicity and toxic side effects.

[0327] 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 the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed within the term "antigen-binding fragment" 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 linked by a disulfide bridge at the hinge region, (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a 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) an isolated complementarity determining region (CDR) or (vii) a combination of two or more isolated CDRs which can optionally be joined by a synthetic linker. Furthermore, although the two domains of the Fv fragment, VL and VH, are coded for by separate genes, they can be joined, using recombinant methods, by a synthetic linker that enables them to be produced as a single protein chain in which the VL and VH regions pair to form monovalent molecules (known as single chain Fv (scFv); see, e.g., 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 encompassed by the term "antigen-binding fragment" of an antibody. Such antibody fragments are obtained using conventional techniques known to those with skill in the art, and are screened for utility in the same fashion as are intact antibodies.

[0328] Fab is an antibody fragment having a molecular weight of about 50,000 and having antigen binding activity, obtained from a fragment of an IgG antibody molecule by treating with a protease, papain, which cleaves the amino acid residue at position 224 of the H chain, in which about one-half of the H chain N-terminal side and the entire L chain are bound together by disulfide bonds.

[0329] F(ab')2 is an antibody fragment having a molecular weight of about 100,000 and having antigen binding activity, obtained by digesting the lower portion of two disulfide bonds in the hinge region of IgG with the enzyme pepsin, and containing two Fab regions connected at the hinge position.

[0330] Fab' is an antibody fragment having a molecular weight of about 50,000 and having antigen binding activity, obtained by cleaving the disulfide bond of the hinge region of the above F(ab')2.

[0331] Further, the Fab' can be produced by inserting a DNA encoding the Fab' fragment of the antibody into an expression vector for prokaryotic expression or eukaryotic expression and introducing the vector into a prokaryotic or eukaryotic to express the Fab'.

[0332] The term "single-chain antibody", "single-chain Fv" or "scFv" means 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 using 1-4 repeats of the variant (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.

[0333] The term "CDR" refers to one of six hypervariable regions within 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 E. A. et al. (1991) Sequences of proteins of immunological interest. NIH Publication 91-3242. As used herein, the Kabat definition of CDRs applies only to CDR1, CDR2 and CDR3 of the light chain variable domain (CDR L1, CDR L2, CDR L3 or L1, L2, L3) and to CDR2 and CDR3 of the heavy chain variable domain (CDR H2, CDR H3 or H2, H3).

[0334] The term "vector" refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. In one embodiment, the vector is a "plasmid," which refers to a circular double stranded DNA loop into which additional DNA segments can be ligated. In another embodiment, the vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome. The vectors disclosed herein are capable of replicating in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors) or can integrate into the genome of the host cell upon introduction into the host cell and be replicated with the host genome (e.g., non-episomal mammalian vectors).

[0335] The term "alkyl" refers to saturated aliphatic hydrocarbon groups, which are straight-chain or branched groups, preferably containing 1 to 20 carbon atoms, more preferably alkyl groups containing 1 to 12 carbon atoms, and even more preferably alkyl groups containing 1 to 10 carbon atoms, and most preferably alkyl groups containing 1 to 6 carbon atoms. The term "Ci-6 alkyl" refers to saturated straight-chain or branched hydrocarbon groups having from 1 to 6 carbon atoms (e.g., 1, 2, 3, 4, 5, or 6 carbon atoms). For example, "Ci-6 alkyl" can be methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, sec-butyl, t-butyl, n-pentyl, i-pentyl, neopentyl, or n-hexyl, and the like.

[0336] The term "cycloalkyl" refers to saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituents, the cycloalkyl ring comprising 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 cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; polycyclic cycloalkyl groups include spiro, fused, and bridged cycloalkyl groups.

[0337] The term "heterocyclyl" refers to saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituents, comprising 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, the heterocyclyl ring comprises 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably, the cycloalkyl ring comprises 3 to 10 ring atoms. Non-limiting examples of monocyclic heterocyclyl groups include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and the like. Polycyclic heterocyclyl groups include spiro, fused, and bridged heterocyclyl groups.

[0338] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (that is, rings which share pairs of adjacent carbon atoms) ring systems having a conjugated pi-electron system, preferably 6- to 10-membered, such as phenyl and naphthyl, preferably phenyl. The aryl ring can be fused to a heteroaryl, heterocyclyl, or cycloalkyl ring, wherein the ring that is attached to the parent structure is the aryl ring.

[0339] The term "heteroaryl" refers to a heteroaromatic system comprising 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. Heteroaryl groups are 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 can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring to which the parent structure is attached together with the heteroaryl ring is an aryl, heterocyclyl or cycloalkyl ring.

[0340] The term "haloalkyl" refers to an alkyl group substituted with one or more halo groups.

[0341] The term "deuteroalkyl" refers to an alkyl group substituted with one or more deuterium atoms.

[0342] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0343] The terms "substituted" and "substitution" mean that one or more (e.g., one, two, three or four) hydrogens on the designated atom is / are replaced with a selection from the indicated group, provided that normal valency 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.

[0344] If a substituent is described as "optionally substituted" with a group, then the substituent can be (1) unsubstituted, or (2) substituted with the group. If an atom or group is described as being optionally substituted with one or more of a list of substituents, then one or more hydrogens on the atom or group are replaced with an independently selected, optional substituent. If a substituent is described as being "independently selected from" or "each independently," then each substituent is selected independently of the others. Thus, each substituent can be the same or different from another (or other) substituent(s). For example, where a substituent or substitution position, or different substituents or substitution positions, have a selection of R groups (such as, but not limited to, R2, R3, Rh, Ri, Rx, and / or Ry) that can be the same or different designations, each R is selected independently, i.e., can be the same or different. The same is true for selections of numerical values such as d, g, m, n.

[0345] Unless indicated otherwise, as used herein, the point of attachment of a substituent can be from any suitable position on the substituent.

[0346] When a bond to a substituent is shown as a dashed line passing through a bond connecting two atoms in a ring, then such substituent can be bonded to either atom in the ring that is replaceable.

[0347] The terms "comprise", "comprising", "have", "having", "include", "including", "contain", "containing", or "involve" and any variations thereof herein are open-ended, and do not exclude additional, unrecited elements or method steps. It will be understood by those within the art that the term "comprising" encompasses the terms "consisting of" and "consisting essentially of".

[0348] A "pharmaceutically acceptable carrier" in the present application means a diluent, adjuvant, excipient, or vehicle with which the active ingredient is administered, and which is suitable for use in contact with the tissues of human beings and / or other animals without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit / risk ratio, within the scope of sound medical judgment. The term "active ingredient", "therapeutic agent", "active substance" or "active agent" means a chemical entity that is effective in treating one or more symptoms of a target disorder or condition.

[0349] The term "effective amount" (e.g., "therapeutically effective amount" or "prophylactically effective amount") as used herein means the amount of active ingredient which will elicit the desired effect, e.g., relief from one or more symptoms of the disorder being treated or prevention of the occurrence of the disorder or its symptoms.

[0350] The term "treat" as used herein means to reverse, alleviate, inhibit, or prevent the progression of the disorder or condition to which such term applies, or to ameliorate symptoms of such disorder or condition. DETAILED DESCRIPTION

[0351] Embodiments of the present application will be described in detail with reference to the following Examples, but the present application should not be construed as being limited thereto. Unless otherwise specified, the conditions in the Examples are those under ordinary conditions or those recommended by the manufacturer. Unless otherwise specified, the reagents or instruments used in the Examples are those commercially available.

[0352] Abbreviations appearing herein and their meanings are as follows:

[0353] Table 1 Abbreviations and their meanings

[0354] Cell name and source

[0355] Unless otherwise specified, the cell lines or cell strains used in the Examples of the present application can be obtained through commercial channels.

[0356] Example 1: Synthesis of Compound E1

[0357] Step 1: Synthesis of compound E1-2

[0358] Compound E1-1 (2.74 g, 15.5 mmol) was dissolved in water (100 ml), sodium carbonate (2.74 g, 25.8 mmol) was added, benzyl chloroformate was dissolved in THF (20 ml), and the mixture was added dropwise to the reaction system under ice bath, and stirred at room temperature for 4 hours. The reaction was monitored by LCMS. The reaction solution was extracted with EA (40 ml*2), and the organic phase was combined and dried over anhydrous sodium sulfate. After concentration, the product was purified by reversed-phase column chromatography to obtain compound E1-2 (3.8 g, 78.7%) as a gray solid. LC-MS (ESI): m / z found [M+H] + = 311.4.

[0359] Step 2: Synthesis of compound E1-3

[0360] Compound E1-2 (1.2 g, 3.87 mmol) was dissolved in ACN (10 ml), sodium carbonate (2.67 g, 19.35 mmol) and 1,2-dibromoethane (3.6 g, 19.35 mmol) were added, and the reaction system was closed and stirred at 80°C for 16 hours. The reaction was monitored by LCMS. The reaction solution was extracted with EA (40 ml*2) and water (60 ml), and the organic phase was combined and dried over anhydrous sodium sulfate. After concentration, the product was purified by normal phase column chromatography to obtain compound E1-3 (700 mg, 43.3%) as a yellow oil. LC-MS (ESI): m / z found [M+H] + = 417.3.

[0361] Step 3: Synthesis of compound E1-4

[0362] Compound E1-3 (700 mg, 1.68 mmol) was dissolved in EtOH (6 ml), methylamine hydrochloride (1.136 g, 16.8 mmol) and DIEA (2.6 g, 20.16 mmol) were added, and the reaction system was closed and stirred at 100°C for 3 hours. The reaction was monitored by LCMS. The reaction solution was concentrated to obtain compound E1-4 (490 mg, 79.3%) as a yellow oil. LC-MS (ESI): m / z found [M+H] + = 368.3.

[0363] Step 4: Synthesis of compound E1-5

[0364] Compound E1-4 (490 mg, 1.33 mmol) was dissolved in DCM (25 ml), and TEA (330 mg, 3.26 mmol) and di-tert-butyl dicarbonate (533 mg, 2.45 mmol) were added. The mixture was stirred at room temperature for 3 hours. The reaction was monitored by LC-MS to indicate completion. The reaction solution was extracted with EA (40 ml * 2) and water (60 ml). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by normal-phase column chromatography to give compound E1-5 (490 mg, 78.7%) as a yellow oil. LC-MS (ESI): m / z found [M+H] + =468.6.

[0365] Step 5: Synthesis of compound E1-6

[0366] Compound E1-5 (490 mg, 1.05 mmol) was dissolved in MeOH (12 mL), and Pd-C (290 mg, 60% wt) was added. The mixture was purged with hydrogen and stirred at room temperature for 16 hours. The reaction was monitored for completion by LC-MS. The reaction solution was filtered, concentrated, and purified by HPLC to obtain compound E1-6 (200 mg, 57.2%) as a white solid. LC-MS (ESI): m / z found [M+H] + =334.4.

[0367] Step 6: Synthesis of compound E1-7

[0368] Compound E1-6 (107 mg, 0.32 mmol) was dissolved in EtOH (2 mL), and sodium acetate (58 mg, 0.7 mmol) and Int.1 (20 mg, 0.032 mmol) were added. The mixture was stirred at 50 °C for 4 hours, then stirred at room temperature for 16 hours. The reaction was monitored by LC-MS to indicate completion. The reaction solution was concentrated and extracted with EA (5 mL x 2) and water (10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by normal-phase column chromatography to give compound E1-7 (20 mg, 66.7%) as a white solid. LC-MS (ESI): m / z found [M+H] + =937.1.

[0369] Step 7: Synthesis of compound E1-8

[0370] Compound E1-7 (20 mg, 0.021 mmol) was dissolved in THF (1 ml), HCl / 1,4-dioxane (0.5 ml, 4M) was added, and stirring was performed at room temperature for 2 hours. The reaction was monitored to completion by LCMS. After the reaction solution was poured into saturated sodium bicarbonate solution (10 ml), extraction was performed with DCM (5 ml*2), the organic phase was combined, dried over anhydrous sodium sulfate, concentrated, and purified by normal phase column chromatography to obtain compound E1-8 (10 mg, 56.9%) as a white solid. LC-MS (ESI): m / z found [M+H]=837.1. +

[0371] Eighth step: synthesis of compound E1-9

[0372] Compound E1-8A (1.3 mg, 0.006 mmol) was dissolved in DMF (1 ml), HOBT (1.58 mg, 0.0066 mmol), DIEA (1.63 mg, 0.012 mmol), and compound 8 (5.2 mg, 0.006 mmol) were added, and stirring was performed at room temperature for 2 hours. The reaction was monitored to completion by LCMS. The reaction solution was purified by reverse phase column chromatography to obtain compound E1-9 (2.5 mg) as a white solid. LC-MS (ESI): m / z found [M+H]=926.02. +

[0373] Example 2: synthesis of compound E2

[0374] First step: synthesis of compound E2-2

[0375] Compound E1-1 (1 g, 3.22 mmol) was dissolved in DMF (20 mL), K2CO3 (887 mg, 6.57 mmol) was added, and then 1-Boc-3-iodazetidine (1.36 g, 4.83 mmol) was added, and reflux was performed at 120°C for 8 hours. The reaction was monitored to completion by LCMS. Extraction was performed with saturated aqueous NaCl solution (60 ml) and EA (40 ml), the organic phase was combined, dried over anhydrous Na2SO4, concentrated under reduced pressure, and purified by normal phase column chromatography (37% EA: PE). Concentration under reduced pressure gave compound E2-2 (886 mg, 59.1%) as a yellowish oil. LC-MS (ESI): m / z found [M+H]=366.2. +

[0376] Second step: synthesis of compound E2-3

[0377] ​​​Compound E2-2 (886 mg, 1.9045 mmol) was dissolved in MeOH (26.6 mL), and Pd / C (270 mg, 60% wt) was added. The reaction was carried out at room temperature for 16 hours under hydrogen protection. The reaction was monitored by LC-MS. The reaction solution was filtered and concentrated, purified by reversed-phase column chromatography, and concentrated under reduced pressure to obtain crude E2-3 (464.7 mg), which was purified by HPLC to obtain a grayish-white solid (104 mg, 16.5%). LC-MS (ESI): m / z found [M+H] + =332.2.

[0378] Step 3: Synthesis of compound E2-4

[0379] Compound E2-3 (63.9 mg, 0.192 mmol) was dissolved in AcOH (4.8 mL), and Int.1 (30 mg, 0.048 mmol) was added. The mixture was stirred at room temperature for 16 hours. The reaction was monitored by LC-MS to indicate completion. The reaction solution was concentrated and extracted with EA (5 mL x 2) and saturated sodium bicarbonate aqueous solution (10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by normal-phase column chromatography to give compound E2-4 (30 mg, 66.8%) as a white solid. LC-MS (ESI): m / z found [M+H] + =935.1.

[0380] Step 4: Synthesis of compound E2

[0381] Compound E2-4 (40 mg, 0.043 mmol) was dissolved in THF (2 ml), and HCl / 1,4-dioxane (1.5 ml, 4 M) was added. The mixture was stirred at room temperature for 3 hours. The reaction was monitored by LC-MS to indicate completion. The reaction solution was poured into a saturated sodium bicarbonate solution (10 ml), extracted with DCM (5 ml * 2), and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by normal-phase column chromatography to give compound E2 (20 mg, 55.7%) as a white solid. LC-MS (ESI): m / z found [M+H] + =835.1.

[0382] Example 3: Synthesis of compound E6

[0383] Step 1: Synthesis of compound E6-2

[0384] Compound E2-2 (1.1 g, 2.36 mmol) was dissolved in THF (10 mL), and HCl / 1,4-dioxane solution (5 mL) was added. The mixture was stirred at room temperature for 5 hours. The reaction was monitored by LC-MS to indicate completion. The reaction solution was poured into a saturated sodium bicarbonate solution (30 mL) and extracted with EA (20 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to give compound E6-2 (630 mg, 73.0%) as a deep yellow solid. LC-MS (ESI): m / z found [M+H] + =365.43.

[0385] Step 2: Synthesis of compound E6-3

[0386] Compound E6-2 (700 mg, 1.90 mmol) was dissolved in DMF (7 mL), and glycolic acid (121.1 mg, 1.59 mmol), HOBt (64.4 mg, 0.476 mmol), EDCI (396.2 mg, 2.065 mmol), and TEA (320.6 mg, 3.18 mmol) were added. The reaction was carried out at room temperature for 2 hours. The reaction was monitored by LC-MS. The reaction solution was purified by reversed-phase column chromatography and concentrated under reduced pressure to give E6-3 (310 mg, 38.5%) as a yellow oil. LC-MS (ESI): m / z found [M+H] + =423.47.

[0387] Step 3: Synthesis of compound E6-4

[0388] Compound E6-3 (300 mg, 0.707 mmol) was dissolved in MeOH (9 mL), and Pd / C (180 mg, 60% wt) was added. The reaction was carried out at room temperature for 16 hours under hydrogen protection. The reaction was monitored by LC-MS to indicate completion. The reaction solution was filtered, concentrated, purified by reversed-phase column chromatography, and then concentrated under reduced pressure. HPLC purification yielded E6-4 (57 mg, 27.8%) as a white solid. LC-MS (ESI): m / z found [M+H] + =289.34.

[0389] Step 4: Synthesis of compound E6

[0390] Compound E6-4 (10 mg, 0.032 mmol) was dissolved in AcOH (0.8 ml), added Int. 1 (5 mg, 0.008 mmol), replaced with nitrogen for three times, stirred at room temperature for 16 hours. LCMS monitored the completion of the reaction. The reaction was purified by HPLC to obtain compound E6 (4.1 mg, 57.4%) as a white solid. LC-MS (ESI): m / z found [M+H]=892.98. + = 892.98.

[0391] Example 4: Synthesis of compound E7

[0392] First step: Synthesis of compound E7-2

[0393] Compound E7-1 (1 g, 4.93 mmol) was dissolved in nitromethane (22 ml), added ammonium acetate (1 g, 13.06 mmol), replaced with nitrogen for three times, stirred at 102°C for 1.5 hours. LCMS monitored the completion of the reaction. The reaction was cooled and filtered, the filter cake was washed with water, dried to obtain compound E7-2 (1.1 g, 90.6%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=246.22. + = 246.22.

[0394] Second step: Synthesis of compound E7-3

[0395] Compound E7-2 (900 mg, 3.64 mmol) was dissolved in DMF (10 mL) and methanol (10 ml), added sodium borohydride (275.2 mg, 7.28 mmol) in three portions under ice bath, reacted at room temperature for 1.5 hours. LCMS monitored the completion of the reaction. The reaction was poured into water (50 ml), added 1M HCl to adjust the pH to 5-6, then extracted with EA (30 ml*2), the organic phase was combined, dried, and the sample was stirred, purified by normal phase column chromatography, concentrated to obtain E7-3 (460 mg, 50.7%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=248.24. + = 248.24.

[0396] Third step: Synthesis of compound E7-4

[0397] Compound E7-3 (100 mg, 0.4 mmol) was dissolved in THF (2 mL) and water (1 ml), lithium hydroxide (84 mg, 2 mmol) was added, and the reaction was stirred at room temperature for 2 hours. LCMS monitoring showed that the reaction was completed. The reaction was poured into water (5 ml), extracted with EA (3 ml*2), the organic phase was combined, dried, and purified by normal phase column chromatography. After concentration, E7-4 (90 mg, 96.0%) was obtained as a yellow solid. LC-MS (ESI): m / z found [M+H] + = 234.21.

[0398] Fourth step: synthesis of compound E7-5

[0399] Compound E7-4 (80 mg, 0.34 mmol) was dissolved in 1,4-dioxane (1 ml), TEA (68.7 mg, 0.68 mmol) and dppa (97 mg, 0.41 mmol) were added, and the reaction was stirred at room temperature for 0.5 hours. E7-4A (63.92 mg, 0.34 mmol) was dissolved in 1,4-dioxane (0.5 ml) and added to the above reaction system. The reaction system was heated to 100°C and stirred for 1 hour. LCMS monitoring showed that the reaction was completed. The reaction was poured into water (5 ml), extracted with EA (3 ml*2), the organic phase was combined, dried and concentrated, and then wet loaded and purified by normal phase column chromatography to obtain compound E7-5 (60 mg, 43.5%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 405.46.

[0400] Fifth step: synthesis of compound E7-6

[0401] Compound E7-5 (60 mg, 0.148 mmol) was dissolved in MeOH (3 ml), Pd-C (42 mg, 70% wt) was added, and hydrogen was replaced. The reaction was stirred at room temperature for 16 hours. LCMS monitoring showed that the reaction was completed. The reaction was filtered and concentrated, and then purified by HPLC to obtain compound E7-6 (16 mg, 27.7%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 389.50.

[0402] Sixth step: synthesis of compound E7-7

[0403] Compound E7-6 (16 mg, 0.041 mmol) was dissolved in AcOH (1 ml), Int. 1 (10 mg, 0.016 mmol) was added, replaced with nitrogen for three times, stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction was concentrated and extracted with EA (5 ml*2) and water (10 ml), the organic phase was combined and dried over anhydrous sodium sulfate, concentrated and purified by normal phase column chromatography to obtain compound E7-7 (9 mg, 56.6%) as a white solid. LC-MS (ESI): m / z found [M+H]=993.15. + +

[0404] Step 7: Synthesis of compound E7

[0405] Compound E7-7 (9 mg, 0.009 mmol) was dissolved in THF (0.5 ml), HCl / 1,4-dioxane (0.25 ml, 4M) was added, stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction was purified by HPLC to obtain compound E7 (2.6 mg, 32.3%) as a white solid. LC-MS (ESI): m / z found [M+H]=893.03. + +

[0406] Example 5: Synthesis of compound E8

[0407] Step 1: Synthesis of compound E8-2

[0408] Compound E7-4 (200 mg, 0.851 mmol) was dissolved in 1,4-dioxane (2 ml), TEA (172 mg, 1.702 mmol) and dppa (280.8 mg, 1.021 mmol) were added, stirred at room temperature for 0.5 hours, then E8-1A (168 mg, 0.851 mmol) was dissolved in 1,4-dioxane (1 ml) and added to the above reaction system, the reaction system was heated to 100°C and stirred for 1 hour. The reaction was monitored by LCMS. The reaction was poured into water (10 ml) and extracted with EA (6 ml*2), the organic phase was combined and dried, concentrated and purified by normal phase column chromatography to obtain compound E8-2 (287 mg, 78.5%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=429.48. + +

[0409] Step 2: Synthesis of compound E8-3

[0410] Compound E8-2 (140 mg, 0.325 mmol) was dissolved in MeOH (4.2 ml), Pd-C (98 mg, 70%wt) was added, hydrogen was replaced, and the mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction was filtered and concentrated, and purified by HPLC to give compound E8-3 (22 mg, 16.9%) as a white solid. LC-MS (ESI): m / z found [M+H]=399.50. +

[0411] Step 3: Synthesis of compound E8-4

[0412] Compound E8-3 (22 mg, 0.055 mmol) was dissolved in AcOH (1 ml), Int.1 (10 mg, 0.016 mmol) was added, and the mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction was concentrated, extracted with EA (5 ml*2) and water (10 ml), and dried over anhydrous sodium sulfate. The mixture was concentrated and purified by normal phase column chromatography to give compound E8-4 (9 mg, 56.6%) as a white solid. LC-MS (ESI): m / z found [M+H]=1003.14. +

[0413] Step 7: Synthesis of compound E8

[0414] Compound E8-4 (9 mg, 0.009 mmol) was dissolved in DCM (0.5 ml), TFA (0.2 ml) was added, and the mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction was purified by HPLC to give compound E8 (3.2 mg, 39.4%) as a white solid. LC-MS (ESI): m / z found [M+H]=903.02. +

[0415] Example 6: Synthesis of compound E9

[0416] Step 1: Synthesis of compound E9-1

[0417] ​​​Compound E2-2 (1.5 g, 3.22 mmol) was dissolved in THF (50 ml), HCl / 1,4-dioxane (10 ml, 4M) was added, and stirred at room temperature for 4 hours. The reaction was monitored by LCMS to be complete. The reaction was extracted with EA (40 ml*2), and the organic phase was combined, dried over anhydrous sodium sulfate, and concentrated to give compound E9-1 (890 mg, 73.9%) as a yellow oil. LC-MS (ESI): m / z found [M+H]=366.8. +

[0418] Second Step: Synthesis of compound E9-2

[0419] Compound E9-1 (150 mg, 0.42 mmol) was dissolved in THF (1 ml), DIEA (63 mg, 0.49 mmol) and triphosgene (60 mg, 0.21 mmol) were added, and stirred at room temperature for 40 minutes. 2-(tert-Butyldimethylsiloxy)ethylamine was dissolved in THF (3 ml), DIEA (159 mg, 1.23 mmol) was added, and it was slowly added dropwise to the above system, and stirred at room temperature for 4 hours. The reaction was monitored by LCMS to be complete. The reaction was extracted with EA (40 ml*2) and water (60 ml), and the organic phase was combined, dried over anhydrous sodium sulfate, and concentrated to give compound E9-2 (67 mg, 28.2.%) as a yellow oil. LC-MS (ESI): m / z found [M+H]=567.6. +

[0420] Third Step: Synthesis of compound E9-3

[0421] Compound E9-2 (67 mg, 1.68 mmol) was dissolved in THF (1.5 ml), tetrabutylammonium chloride (0.3 ml, 1M) was added, and stirred at room temperature for 4 hours. The reaction was monitored by LCMS to be complete. The reaction was concentrated and purified by reverse phase column chromatography to give compound E9-3 (26 mg, 48.7%) as a light yellow oil. LC-MS (ESI): m / z found [M+H]=453.2. +

[0422] Fourth Step: Synthesis of compound E9-4

[0423] ​​​Compound E9-3 (26 mg, 0.06 mmol) was dissolved in MeOH (1.5 mL), and Pd / C (15.6 mg, 60% wt) was added. The mixture was purged with hydrogen and stirred at room temperature for 16 hours. The reaction was monitored for completion by LC-MS. The reaction solution was filtered, concentrated, and purified by HPLC to obtain compound E9-4 (10 mg, 52.4%) as a white solid. LC-MS (ESI): m / z found [M+H] + =319.2.

[0424] Step 5: Synthesis of compound E9

[0425] Compound E9-4 (10 mg, 0.03 mmol) was dissolved in HoAC (1.5 mL), and Int.1 (5 mg, 0.08 mmol) was added. The mixture was stirred at room temperature for 16 hours. The reaction was monitored for completion by LC-MS. The reaction solution was filtered, concentrated, and purified by HPLC to give compound E9 (1.92 mg, 7%) as a white solid. LC-MS (ESI): m / z found [M+H] + =922.2.

[0426] Example 7: Synthesis of compound E10

[0427] Step 1: Synthesis of compound E10-2

[0428] Compound E7-4 (110 mg, 0.46 mmol) was dissolved in water (2 ml), and TEA (0.13 ml, 0.94 mmol) and DPPA (0.12 ml, 0.56 mmol) were added. The mixture was reacted at room temperature for 30 min, followed by the addition of tert-butyl(2-aminoethyl)(methyl)carbamate (245 mg, 1.40 mmol), and the temperature was raised to 100 °C for 2 h. The reaction was monitored by LC-MS to ensure complete reaction. The reaction solution was cooled to room temperature and extracted with EA. The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by normal-phase column chromatography to give compound E10-2 (83 mg, 43.7%) as a pale yellow oil. LC-MS (ESI): m / z found [M+H] + =405.2.

[0429] Step 2: Synthesis of compound E10-3

[0430] Compound E10-2 (158 mg, 0.39 mmol) was dissolved in MeOH (5 ml), palladium on carbon (110.6 mg) was added and hydrogen was bubbled into the solution. The reaction was stirred at room temperature overnight. LCMS was used to monitor the completion of the reaction. The reaction mixture was filtered through celite and the filtrate was concentrated and purified by reverse phase column chromatography to give compound E10-3 (89 mg, 60.9%) as a white solid. LC-MS (ESI): m / z found [M+H]=375.5. +

[0431] Step 3: Synthesis of compound E10-4

[0432] Compound E10-3 (24 mg, 0.064 mmol) was dissolved in AcOH (1 ml), Int.1 (10 mg, 0.016 mmol) was added and the reaction was stirred at room temperature overnight. LCMS was used to monitor the completion of the reaction. The reaction mixture was adjusted to pH = 7 with a saturated aqueous sodium bicarbonate solution and extracted with EA (5 ml*2) and water (10 ml). The organic phase was combined, dried over anhydrous sodium sulfate, concentrated and purified by normal phase column chromatography to give compound E10-4 (11 mg, 70.06%) as a white solid. LC-MS (ESI): m / z found [M+H]=979.1. +

[0433] Step 4: Synthesis of compound E10

[0434] Compound E10-4 (11 mg, 0.011 mmol) was dissolved in THF (1 ml), HCl / 1,4-dioxane (0.5 ml, 4M) was added and the reaction was stirred at room temperature for 2 hours. LCMS was used to monitor the completion of the reaction. The reaction mixture was poured into a saturated sodium bicarbonate solution (10 ml) and extracted with DCM (5 ml*2). The organic phase was combined, dried over anhydrous sodium sulfate, concentrated and purified by normal phase column chromatography to give compound E10 (4.72 mg, 56.9%) as a white solid. LC-MS (ESI): m / z found [M+H]=879.0. +

[0435] Example 8: Synthesis of compound E11

[0436] Step 1: Synthesis of compound E11-2

[0437] ​​​Compound E7-4 (300 mg, 1.28 mmol) was dissolved in dioxane (5 ml), TEA (0.35 ml, 2.55 mmol) and DPPA (0.34 ml, 1.53 mmol) were added, and the reaction was allowed to proceed at room temperature for 30 min. 2-(tert-Butyldimethylsilyl)oxy)ethan-1-amine (224 mg, 1.28 mmol) was added, and the reaction was allowed to proceed at 100 °C for 2 h. The reaction was monitored by LCMS. The reaction was allowed to cool to room temperature, and the reaction was extracted with EA. The organic phase was combined and dried over anhydrous sodium sulfate. The solvent was removed by concentration, and the residue was purified by normal phase column chromatography to give compound E1-2 (152 mg, 29.3%) as a white solid. LC-MS (ESI): m / z found [M+H]+= 406.6.

[0438] Second Step: Synthesis of compound E11-3

[0439] Compound E11-2 (202 mg, 0.50 mmol) was dissolved in THF (2.5 mL), and TBAF (1 mL, 1.49 mmol, 1M in THF) was added. The reaction was allowed to proceed at room temperature for 3 h under nitrogen. The reaction was monitored by LCMS. The reaction was filtered and concentrated. The residue was purified by reverse phase column chromatography and concentrated under reduced pressure to give a yellowish solid (120 mg, 82.8%). LC-MS (ESI): m / z found [M+H]+= 292.3. +

[0440] Third Step: Synthesis of compound E11-4

[0441] Compound E11-3 (120 mg, 0.41 mmol) was dissolved in MeOH (5 ml), and palladium on carbon (84 mg) was added. Hydrogen was introduced, and the reaction was allowed to proceed at room temperature overnight. The reaction was monitored by LCMS. The reaction was filtered through celite, and the filtrate was concentrated and purified by reverse phase column chromatography to give compound E11-4 (34 mg, 31.5%) as a white solid. LC-MS (ESI): m / z found [M+H]+= 262.3. +

[0442] Fourth Step: Synthesis of compound E11

[0443] ​​Compound E11-4 (8.5 mg, 0.323 mmol) was dissolved in AcOH (0.5 ml), Int.1 (5 mg, 0.008 mmol) was added, stirred at room temperature for 16 hours. LCMS monitoring reaction was complete. The reaction was concentrated and extracted with EA (5 ml*2) and saturated aqueous sodium bicarbonate solution (10 ml), the organic phase was combined and dried over anhydrous sodium sulfate, concentrated and purified by normal phase column chromatography to give compound E11 (0.52 mg, 7.4%) as a white solid. LC-MS (ESI): m / z found [M+H]=866.0 + = 866.0

[0444] Example 9: Synthesis of compound E12

[0445] First step: Synthesis of compound E12-1

[0446] Compound E12-1 (10 g, 15.5 mmol) was dissolved in Et2O (100 ml), oxalyl chloride (2.74 g, 24.7 mmol) was added, refluxed overnight. LCMS monitoring reaction was complete. The reaction was filtered, washed with ether, NH4.MeOH (15 ml) was added to the solid, stirred at room temperature for 4 h, the reaction was concentrated and purified by normal phase column to give compound E12-2 (3.8 g, 26.6%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=234.1. + = 234.1.

[0447] Second step: Synthesis of compound E12-3

[0448] Compound E12-2 (2 g, 3.87 mmol) was dissolved in MeOH (20 ml), Pd-C (200 mg, 60% wt) was added, hydrogen was replaced, stirred at room temperature for 16 hours. LCMS monitoring reaction was complete. The reaction was filtered and concentrated, purified by HPLC to give compound E12-3 (1.54 g, 88.5%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=204.2. + = 204.2.

[0449] Third step: Synthesis of compound E12-4

[0450] Compound E12-3 (1.54 g, 1.33 mmol) was dissolved in DCM (40 ml), TEA (2.29 g, 3.99 mmol) and di-tert-butyl dicarbonate (3.3 g, 2.66 mmol) were added, stirred at room temperature for 2 hours. LCMS monitoring reaction was completed. The reaction was extracted with EA (40 ml*2) and water (60 ml), the organic phase was combined and dried over anhydrous sodium sulfate, concentrated and purified by normal phase column chromatography to give compound E12-4 (1.3 g, 56.7%) as a yellow oil. LC-MS (ESI): m / z found [M+H] = 304.1. +

[0451] Fourth step: synthesis of compound E12-5

[0452] Compound E12-4 (500 mg, 3.30 mmol) was dissolved in dioxane (25 ml), lithium aluminum hydride (6.6 ml, 2.5 mol in THF) was added, and the reaction was refluxed overnight. LCMS monitoring reaction was completed. H2O (2.5 ml), 15% NaOH aqueous solution (2.5 ml), H2O (7.5 ml) were added dropwise under ice water bath, the ice water bath was removed, stirred at room temperature for 1 hour, filtered with diatomite, the filtrate was concentrated under reduced pressure, concentrated and purified by HPLC to give compound E12-5 (200 mg, 64.3%) as a yellow oil. LC-MS (ESI): m / z found [M+H] = 190.2. +

[0453] Fifth step: synthesis of compound E12-6

[0454] Compound E12-5 (10 mg, 0.32 mmol) was dissolved in EtOH (2 ml), sodium acetate (58 mg, 0.7 mmol) and Int. 1 (20 mg, 0.032 mmol) were added, stirred at room temperature for 6 hours. LCMS monitoring reaction was completed. The reaction was concentrated and extracted with EA (5 ml*2) and water (10 ml), the organic phase was combined and dried over anhydrous sodium sulfate, concentrated and purified by normal phase column chromatography to give compound E12-6 (16 mg, 66.7%) as a white solid. LC-MS (ESI): m / z found [M+H] = 793.9. +

[0455] Sixth step: synthesis of compound E12

[0456] ​​​Compound E12-6 (5 mg, 0.063 mmol) was dissolved in DMF (2 ml), 2- hydroxycyclobutane-1 -carboxylic acid (3.66 mg, 0.315 mmol), HATU (3.59 mg, 0.095 mmol), DIEA (8.13 mg, 0.63 mmol) were added and stirred at room temperature for 1 h. LCMS was used to monitor the completion of the reaction. To the reaction mixture was added EA (10 ml), the mixture was washed with saturated sodium bicarbonate (20 ml), the organic phase was dried, filtered, concentrated and purified by HPLC to give compound E12 (1.5 mg, 26.7%) as a white solid. LC-MS (ESI): m / z found [M+H]=892.0. +

[0457] Example 10: Synthesis of compound E13

[0458] First Step: Synthesis of compound E13-2

[0459] Compound E7-3 (500 mg, 3.30 mmol) was dissolved in THF (25 ml), lithium aluminum hydride (12.1 ml, 2.5 mol in THF) was added and stirred at room temperature overnight. LCMS was used to monitor the completion of the reaction. H2O (2.5 ml), 15% NaOH aqueous solution (2.5 ml), H2O (7.5 ml) were added dropwise successively under ice water bath cooling, the ice water bath was removed and stirred at room temperature for 1 h, filtered with diatomite, the filtrate was concentrated under reduced pressure, concentrated and purified by HPLC to give compound E13-2 (150 mg, 39.2%) as a yellow oil. LC-MS (ESI): m / z found [M+H]=191.2. +

[0460] Second Step: Synthesis of compound E13-3

[0461] Compound E13-2 (12.2 mg, 1.28 mmol) was dissolved in AcOH (2 ml), Int. 1 (10 mg, 0.32 mmol) was added and stirred at room temperature for 6 h. LCMS was used to monitor the completion of the reaction. The reaction mixture was concentrated and extracted with EA (5 ml*2) and saturated sodium bicarbonate aqueous solution (10 ml), the organic phase was combined, dried with anhydrous sodium sulfate, concentrated and purified by HPLC to give compound E13 (5.97 mg, 47.1%) as a white solid. LC-MS (ESI): m / z found [M+H]=794.8. +

[0462] Example 11: Synthesis of compound E14 ​​​

[0463] First Step: Synthesis of compound E14-2

[0464] Compound E7-4 (200 mg, 0.851 mmol) was dissolved in 1,4-dioxane (2 ml), TEA (172 mg, 1.702 mmol) and dppa (280.8 mg, 1.021 mmol) were added, after stirring at room temperature for 0.5 hours, E14-1A (146.37 mg, 0.851) was dissolved in 1,4-dioxane (1 ml) and added to the above reaction system, and the reaction system was warmed to 100 °C and stirred for 1 hour. The reaction was monitored by LCMS. The reaction liquid was poured into water (10 ml) and extracted with EA (6 ml*2), the organic phase was combined and dried and concentrated, and was subjected to normal phase column chromatography for purification to obtain compound E14-2 (280 mg, 81.5%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + = 403.44.

[0465] Second Step: Synthesis of compound E14-3

[0466] Compound E14-2 (140 mg, 0.347 mmol) was dissolved in MeOH (4.2 ml), Pd-C (98 mg, 70% wt) was added, and hydrogen replacement was performed, and stirring was performed at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction liquid was filtered and concentrated, and HPLC purification was performed to obtain compound E14-3 (24 mg, 18.5%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 373.46.

[0467] Third Step: Synthesis of compound E14-4

[0468] Compound E14-3 (24 mg, 0.064 mmol) was dissolved in AcOH (1 ml), Int.1 (10 mg, 0.016 mmol) was added, and nitrogen replacement was performed three times, and stirring was performed at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction liquid was concentrated and extracted with EA (5 ml*2) and water (10 ml), the organic phase was combined and dried with anhydrous sodium sulfate, concentrated and purified by normal phase column chromatography to obtain compound E14-4 (9 mg, 57.5%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 977.10.

[0469] Seventh Step: Synthesis of compound E14

[0470] Compound E14-4 (9 mg, 0.009 mmol) was dissolved in DCM (0.5 ml), TFA (0.2 ml) was added, and stirred at room temperature for 2 hours. The reaction was monitored by LCMS to be completed. The reaction was purified by HPLC to give compound E14 (3.21 mg, 40.5%) as a white solid. LC-MS (ESI): m / z found [M+H] 876.99. + = 876.99.

[0471] Example 12: Synthesis of compound E16

[0472] First Step: Synthesis of compound E16-2

[0473] Compound E16-1 (3 g, 1.5 mmol) was dissolved in DMF (80 ml), potassium carbonate (3.6 g, 1.8 mmol), tert-butyl (2-bromoethyl)carbamate (5.84 g, 1.8 mmol) and sodium iodide (1.08 g, 0.45 mmol) were added, and reacted at 60°C for 4 hours. The reaction was monitored by LCMS to be completed. Extraction was performed with saturated aqueous NaCl solution (60 ml) and EA (40 ml), and the organic phase was combined, dried over anhydrous Na2SO4, concentrated under reduced pressure, and purified by reverse phase column to give compound E16-2 (3.5 g, 57.4%) as a yellow solid. LC-MS (ESI): m / z found [M+H] 372.4. + = 372.4.

[0474] Second Step: Synthesis of compound E16-3

[0475] Compound E16-2 (3.5 g, 3.87 mmol) was dissolved in DMF (20 ml), potassium carbonate (5.16 g, 11.61 mmol) and benzyl bromide (2.34 g, 4.257 mmol) were added, and stirred at room temperature for 3 hours. The reaction was monitored by LCMS to be completed. Extraction was performed with saturated aqueous NaCl solution (60 ml) and EA (50 ml), and the organic phase was combined, dried over anhydrous Na2SO4, concentrated under reduced pressure, and purified by normal phase column to give compound E16-3 (3.4 g, 73.6%) as a yellow solid. LC-MS (ESI): m / z found [M+H] 372.4. + = 372.4.

[0476] Third Step: Synthesis of compound E16-4

[0477] Compound E16-3 (500 mg, 0.33 mmol) was dissolved in DMF (8 ml), NaH (75 mg, 0.495 mmol) was added, and the mixture was stirred in an ice bath for 0.5 h. Iodomethane (375 mg, 0.66 mmol) was added dropwise into the system, and the mixture was stirred at room temperature for 2 h. LCMS monitoring showed that the reaction was completed. The reaction solution was extracted with EA (40 ml*2) and water (60 ml). The organic phase was combined, dried over anhydrous sodium sulfate, concentrated, and purified by normal phase column chromatography to obtain compound E16-4 (410 mg, 78.8%) as a yellow oil. LC-MS (ESI): m / z found [M+H]=386.4. +

[0478] Fourth step: synthesis of compound E16-5

[0479] Compound E16-4 (320 mg, 3.30 mmol) was dissolved in nitromethane (5 ml), and ammonium acetate (256 mg, 13.2 mmol) was added. The reaction was refluxed overnight. LCMS monitoring showed that the reaction was completed. The reaction solution was concentrated under reduced pressure, and the concentrate was purified by normal phase to obtain compound E16-5 (340 mg, 92.3%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=429.4. +

[0480] Fifth step: synthesis of compound E16-6

[0481] Compound E16-5 (340 mg, 1.30 mmol) was dissolved in THF (20 ml), and lithium aluminum hydride (1.6 ml, 2.5 mol in THF) was added. The reaction was stirred at room temperature for 2 h. LCMS monitoring showed that the reaction was completed. H2O (2.5 ml), 15% NaOH aqueous solution (2.5 ml), and H2O (7.5 ml) were added dropwise in sequence under ice water bath cooling. The ice water bath was removed, and the mixture was stirred at room temperature for 1 h. The mixture was filtered through diatomite, and the filtrate was concentrated under reduced pressure. The concentrate was purified by HPLC to obtain compound E16-6 (50 mg, 21%) as a yellow oil. LC-MS (ESI): m / z found [M+H]=301.4. +

[0482] Sixth step: synthesis of compound E16-7

[0483] ​​​Compound E16-6 (50 mg, 1.90 mmol) was dissolved in MeOH (26.6 mL) and Pd / C (20 mg, 60% wt) was added, hydrogen was replaced, and the reaction was stirred at room temperature for 16 hours. LCMS monitoring showed the reaction was completed. The reaction was filtered and concentrated to get the crude E16-7 (25 mg). LC-MS (ESI): m / z found [M+H] + = 211.2.

[0484] Step 7: Synthesis of compound E16-8

[0485] Compound E16-7 (25 mg, 0.119 mmol) was dissolved in EtOH (2 ml), Int. 1 (20 mg, 0.032 mmol) and sodium acetate (25 mg, 0.192 mmol) were added, and the reaction was stirred at room temperature for 16 hours. LCMS monitoring showed the reaction was completed. The reaction was concentrated and extracted with EA (5 ml*2) and saturated aqueous sodium bicarbonate solution (10 ml), and the organic phase was combined and dried over anhydrous sodium sulfate, concentrated and purified by HPLC to give compound E16-8 (15 mg, 57.3%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 814.9.

[0486] Step 8: Synthesis of compound E16-10

[0487] Compound E16-9 (200 mg, 1.136 mmol) was dissolved in DCM (8 ml), bis(4-nitrophenyl) carbonate (345.5 mg, 1.136 mmol), TEA (172.2 mg, 1.704 mmol) were added, and the reaction was stirred at room temperature for 2 hours. LCMS monitoring showed the reaction was completed. The reaction was purified by normal phase column chromatography to give compound E16-10 (375 mg, 96.8%) as a colorless oil. LC-MS (ESI): m / z found [M+H] + = 342.4.

[0488] Step 9: Synthesis of compound E16-11

[0489] Compound E16-10 (8.4 mg, 0.024 mmol) was dissolved in DMF (2 ml), HOBT (3.7 mg, 0.027 mmol), DIEA (6.35 mg, 0.049 mmol) and compound E16-8 (15 mg, 0.018 mmol) were added, stirred at room temperature for 2 hours. LCMS monitoring reaction completion. The reaction was purified by HPLC to give compound E16-11 (12 mg) as a white solid. LC-MS (ESI): m / z found [M+H] + = 1017.2.

[0490] Step 10: Synthesis of compound E16

[0491] E16-11 (12 mg, 0.011 mmol) was dissolved in THF (1 ml), TBAF (0.04 ml, 0.023 mmol) was added, stirred at room temperature for 2 hours. LCMS monitoring reaction completion. The reaction was purified by Prep-HPLC to give compound E16 (8.22 mg, 77.2%) as a white solid. LC-MS (ESI): m / z found [1 / 2M+H] + = 902.9.

[0492] Example 13: Synthesis of compound E18

[0493] Step 1: Synthesis of compound E18

[0494] Compound E13 (5.4 mg, 0.045 mmol) was dissolved in ACN (3 ml) and H2O (2 ml), silver nitrate (21.97 mg, 0.855 mmol) was added, stirred at room temperature for 12 hours. LCMS monitoring reaction completion. To the reaction solution, saturated brine (2.5 ml) and saturated sodium bicarbonate (2.5 ml) were added, stirred for 15 min, the mixture was extracted with EA (5 ml*2) and water (10 ml), the organic phase was combined and dried over anhydrous sodium sulfate, concentrated and purified by HPLC to give compound E18 (2 mg, 37.5%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 785.8.

[0495] Example 14: Synthesis of compound E19

[0496] Step 1: Synthesis of compound E19-2

[0497] Compound E19-1 (300 mg, 1.28 mmol) was dissolved in HCl / 1,4-dioxane (2 ml), TEA (259 mg, 2.56 mmol) and DPPA (413 mg, 1.5 mmol) were added, compound 3-aminoazetidine-1-carboxylic acid tert-butyl ester was dissolved in HCl / 1,4-dioxane (2 ml) and added dropwise to the above reaction system, stirred at 120 °C for 6 hours. LCMS monitoring reaction completion. Extracted with saturated NaCl aqueous solution (60 ml) and EA (40 ml), the organic phase was combined and dried over anhydrous Na2SO4, concentrated under reduced pressure, purified by normal phase column chromatography to obtain compound E19-2 (463 mg, 89.8%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 304.1.

[0498] Second step: synthesis of compound E19-3

[0499] Compound E19-2 (245 mg, 0.61 mmol) was dissolved in DCM (5 ml), TFA (2.5 ml) was added, stirred at room temperature for 8 hours. LCMS monitoring reaction completion. The reaction liquid was purified by reversed phase column chromatography to obtain compound E19-3 (148 mg, 79.8%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 304.2.

[0500] Third step: synthesis of compound E19-4

[0501] Compound E19-3 (138 mg, 0.45 mmol) was dissolved in DMF (1 ml), HOBT (13.8 mg, 0.11 mmol), EDCI (94 mg, 0.49 mmol), TEA (115 mg, 1.13 mmol) and 2-hydroxyacetic acid (27.6 mg, 0.38 mmol) were added, stirred at room temperature for 2 hours. LCMS monitoring reaction completion. The reaction liquid was purified by reversed phase column chromatography to obtain compound E19-4 (117 mg, 71.8%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 362.2.

[0502] Fourth step: synthesis of compound E19-5

[0503] Compound E19-5 (17 mg, 0.05 mmol) was dissolved in AcOH (1 ml), Int. 1 (8 mg, 0.01 mmol) was added, stirred at room temperature for 16 hours. LCMS monitored the completion of the reaction. The reaction was filtered and concentrated, HPLC purification to give compound E19 (3.28 mg, 35.1%) as a white solid. LC-MS (ESI): m / z found [M+H]+= 934.6.

[0504] Fifth step: synthesis of compound E19

[0505] Compound E19-5 (17 mg, 0.05 mmol) was dissolved in AcOH (1 ml), Int. 1 (8 mg, 0.01 mmol) was added, stirred at room temperature for 16 hours. LCMS monitored the completion of the reaction. The reaction was filtered and concentrated, HPLC purification to give compound E19 (3.28 mg, 35.1%) as a white solid. LC-MS (ESI): m / z found [M+H]+= 934.6.

[0506] Example 15: synthesis of compound E20

[0507] First step: synthesis of compound E20-1

[0508] Compound E9-1 (400 mg, 1.10 mmol) was dissolved in THF (4 ml), DIEA (169 mg, 1.32 mmol) and triphosgene (160 mg, 0.55 mmol) were added, stirred at room temperature for 40 minutes. tert-Butyl methyl(2-(methylamino)ethyl)carbamate was dissolved in THF (4 ml), DIEA (424 mg, 3.29 mmol) was added, and added dropwise to the above reaction system, stirred at room temperature for 4 hours. LCMS monitored the completion of the reaction. Extracted with saturated aqueous NaCl solution (60 ml) and EA (40 ml), the organic phase was combined, dried over anhydrous Na2SO4, concentrated under reduced pressure, purified by normal phase column chromatography to give compound E20-1 (147 mg, 23.1%) as a colorless liquid. LC-MS (ESI): m / z found [M+H] + = 580.2.

[0509] Second step: synthesis of compound E20-2

[0510] Compound E20-1 (147 mg, 0.25 mmol) was dissolved in MeOH (5 ml), Pd / C (147 mg, 100% wt) was added, hydrogen was replaced, stirred at room temperature for 16 hours. LCMS monitored the reaction was completed. The reaction was filtered and concentrated, HPLC purification to get compound E20-2 (23 mg, 20.6%) as a white solid. LC-MS (ESI): m / z found [M+H]+= 446.2.

[0511] Step 3: Synthesis of compound E20-3

[0512] Compound E20-2 (23 mg, 0.05 mmol) was dissolved in AcOH (2 ml), Int. 1 (10 mg, 0.02 mmol) was added, stirred at room temperature for 16 hours. LCMS monitored the reaction was completed. The reaction was filtered and concentrated, HPLC purification to get compound E20-3 (8 mg, 15.3%) as a white solid. LC-MS (ESI): m / z found [M+H]+= 1048.6.

[0513] Step 4: Synthesis of compound E20

[0514] Compound E20-3 (8 mg, 0.007 mmol) was dissolved in DCM (0.5 ml), TFA (0.2 ml) was added, stirred at room temperature for 2 hours. LCMS monitored the reaction was completed. The reaction was filtered and concentrated, HPLC purification to get compound E20 (5.12 mg, 71.4%) as a white solid. LC-MS (ESI): m / z found [M+H]+= 949.6.

[0515] Example 16: Synthesis of compound E21

[0516] Step 1: Synthesis of compound E21

[0517] Compound E12-6 (10 mg, 0.063 mmol) was dissolved in DMF (2 ml), added (1s,3s)-3-hydroxycyclobutane-1 -carboxylic acid (7.3 mg, 0.315 mmol), HATU (7.2 mg, 0.095 mmol), DIEA (16.3 mg, 0.63 mmol) and stirred at room temperature for 1 h. The reaction was monitored by LCMS. To the reaction was added EA (10 ml), washed with saturated sodium bicarbonate (20 ml), the organic phase was dried, filtered, concentrated and purified by HPLC to give compound E21 (5.8 mg, 51.8%) as a white solid. LC-MS (ESI): m / z found [M+H] = 892.0. +

[0518] Example 17: Synthesis of compound E22

[0519] Step 1: Synthesis of compound E22-2

[0520] Compound E12-6 (10 mg, 0.063 mmol) was dissolved in DMF (2 ml), added 1- (hydroxymethyl)cyclopropane-1 -carboxylic acid (7.3 mg, 0.315 mmol), HATU (7.2 mg, 0.095 mmol), DIEA (16.3 mg, 0.63 mmol) and stirred at room temperature for 1 h. The reaction was monitored by LCMS. To the reaction was added EA (10 ml), washed with saturated sodium bicarbonate (20 ml), the organic phase was dried, filtered, concentrated and purified by HPLC to give compound E22 (2.65 mg, 23.7%) as a white solid. LC-MS (ESI): m / z found [M+H] = 892.0. +

[0521] Example 18: Synthesis of compound E23

[0522] Step 1: Synthesis of compound E23-2

[0523] ​​The DMF (22 ml) was cooled in an ice bath, purged three times with nitrogen, and phosphorus oxychloride (6.25 g, 41.15 mmol) was added. After stirring at room temperature for 15 minutes, compound E23-1 (1 g, 4.115 mmol) was dissolved in DMF (5 ml) and added to the reaction system. After stirring at room temperature for 1 hour, the mixture was heated to 40°C and stirred for 2 hours. The reaction was monitored by LC-MS to indicate completion. The reaction solution was cooled and poured into water (50 ml). The pH was adjusted to alkaline with saturated sodium hydroxide solution. After filtration and drying, compound E23-2 (670 mg, 60.0%) was obtained as a yellow solid. LC-MS (ESI): m / z found [M+H] + =271.06.

[0524] Step 2: Synthesis of compound E23-3

[0525] Compound E23-2 (670 mg, 2.47 mmol) was dissolved in nitromethane (7 mL), and ammonium acetate (504 mg, 6.54 mmol) was added. The mixture was purged with nitrogen three times and refluxed at 102 °C with stirring for 1.5 hours. The reaction was monitored by LC-MS to indicate completion. After cooling, the reaction solution was filtered, the filter cake was washed with water, and dried to obtain compound E23-3 (425 mg, 54.8%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + =314.08.

[0526] Step 3: Synthesis of compound E23-4

[0527] Compound E23-3 (425 mg, 1.35 mmol) was dissolved in DMF (2.5 mL) and methanol (2.5 mL). Sodium borohydride (103 mg, 2.7 mmol) was added in three portions under ice bath conditions, and the reaction was allowed to proceed at room temperature for 1.5 hours. The reaction was monitored by LC-MS to ensure completion. The reaction solution was poured into water (10 mL), and the pH was adjusted to 5-6 with 1 M HCl. Extraction was performed using EA (6 mL x 2). The organic phases were combined, dried, and purified by normal-phase column chromatography. After concentration, E23-4 (280 mg, 65.6%) was obtained as a yellow solid. LC-MS (ESI): m / z found [M+H] + =316.10.

[0528] Step 4: Synthesis of compound E23-5

[0529] Compound E23-4 (140 mg, 0.44 mmol) was dissolved in 1,4-dioxane (3 ml), TEA (0.3 ml), E23-4A (112.2 mg, 0.66 mmol), copper iodide (25 mg, 0.132 mmol) and dichlorobis(triphenylphosphine)palladium (31 mg, 0.044 mmol) were added, and the reaction was stirred at room temperature for 3 hours under nitrogen. The reaction was monitored by LCMS. The reaction was poured into water (8 ml) and extracted with EA (5 ml*2). The organic phase was combined and dried and concentrated. Purification was performed by normal phase column chromatography to obtain compound E23-5 (150 mg, 95.1%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=358.51. +

[0530] Step 5: Synthesis of compound E23-6

[0531] Compound E23-5 (150 mg, 0.419 mmol) was dissolved in MeOH (4.5 ml), Pd-C (500 mg, 300% wt) was added, and the reaction was stirred at room temperature for 16 hours under hydrogen. The reaction was monitored by LCMS. The reaction was filtered and concentrated to obtain compound E23-6 (60 mg, 43.0%) as a white solid. LC-MS (ESI): m / z found [M+H]=332.56. +

[0532] Step 6: Synthesis of compound E23-7

[0533] Compound E23-6 (60 mg, 0.18 mmol) was dissolved in THF (1 ml), TBAF (0.5 ml) was added, and the reaction was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction was purified by HPLC to obtain compound E23-7 (8 mg, 20.3%) as a white solid. LC-MS (ESI): m / z found [M+H]=218.30. +

[0534] Step 7: Synthesis of compound E23

[0535] Compound E23-7 (8 mg, 0.0366 mmol) was dissolved in AcOH (1 ml), Int.1 (5 mg, 0.008 mmol) was added, and the reaction was stirred at room temperature for 16 hours under nitrogen. The reaction was monitored by LCMS. The reaction was purified by HPLC to obtain compound E23 (2.1 mg, 31.9%) as a white solid. LC-MS (ESI): m / z found [M+H]​​​+ = 821.95.

[0536] Example 19: Synthesis of compound E24

[0537] First Step: Synthesis of compound E24-2

[0538] Compound E24-1 (100 mg, 0.15 mmol) was dissolved in ETOH (2.5 ml) and water (2.5 ml), lithium hydroxide (64.62 mg, 0.3 mmol) was added, stirred at room temperature for 4 hours. TLC monitoring reaction complete. The reaction was adjusted to PH with dilute hydrochloric acid, extracted with EA (10 ml*2), the organic phase was combined and dried over anhydrous sodium sulfate, concentrated to give compound E24-2 (15 mg, 78.7%) as a colorless oil. LC-MS (ESI): m / z found [M+H] + = 117.0.

[0539] Second Step: Synthesis of compound E24

[0540] Compound E12-6 (10 mg, 0.063 mmol) was dissolved in DMF (2 ml), (1R, 2R)-2- (hydroxymethyl)cyclopropane-1-carboxylic acid (7.3 mg, 0.315 mmol), HATU (7.2 mg, 0.095 mmol), DIEA (16.3 mg, 0.63 mmol) were added, stirred at room temperature for 1 hour. LCMS monitoring reaction complete. To the reaction was added EA (10 ml), saturated sodium bicarbonate (20 ml) was washed, the organic phase was dried, filtered, rotary evaporated, concentrated and purified by HPLC to give compound E24 (8.35 mg, 74.6%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 891.3.

[0541] Example 20: Synthesis of compound E26

[0542] First Step: Synthesis of compound E26

[0543] Compound E12-6 (10 mg, 0.012 mmol) was dissolved in DMF (2 ml), 1- hydroxycyclopropane-1-carboxylic acid (6.43 mg, 0.063 mmol), HATU (7.1 mg, 0.018 mmol), DIEA (16.3 mg, 0.12 mmol) were added and stirred at room temperature for 1 hour. The reaction was monitored to completion by LCMS. To the reaction solution was added EA (10 ml), saturated sodium bicarbonate (20 ml) was added for washing, the organic phase was dried, filtered, concentrated and purified by HPLC to give compound E26 (3.2 mg, 28.9%) as a white solid. LC-MS (ESI): m / z found [M+H]=877.9. +

[0544] Example 21: Synthesis of compound E28

[0545] Step 1: Synthesis of compound E28

[0546] Compound E21 (4.8 mg, 0.005 mmol) was dissolved in ACN (3 ml) and H2O (2 ml), silver nitrate (17.4 mg, 0.102 mmol) was added and stirred at room temperature for 12 hours. The reaction was monitored to completion by LCMS. To the reaction solution was added saturated brine (2.5 ml) and saturated sodium bicarbonate (2.5 ml), stirred for 15 min, the mixture was extracted with EA (5 ml*2) and water (10 ml), the organic phase was combined and dried over anhydrous sodium sulfate, concentrated and purified by HPLC to give compound E28 (0.85 mg, 17.8%) as a white solid. LC-MS (ESI): m / z found [M+H]=865.0. +

[0547] Example 22: Synthesis of compound E29

[0548] Step 1: Synthesis of compound E29-2

[0549] Compound E29-1 (1 g, 5.3 mmol) was dissolved in DMF (80 ml), potassium carbonate (2.2 g, 15.9 mmol) and 1,2-2 bromoethane (1.2 g, 6.36 mmol) were added and reacted at 120 °C for 16 hours. The reaction was monitored to completion by LCMS. Extraction was performed with saturated aqueous NaCl solution (60 ml) and EA (40 ml), the organic phase was combined and dried over anhydrous Na2SO4, concentrated under reduced pressure and purified by normal phase column to give compound E29-2 (1.1 g, 96.7%) as a yellow solid. LC-MS (ESI): m / z found [M+H]​​+ = 215.0.

[0550] Second Step: Synthesis of compound E29-3

[0551] Compound E29-2 (1.1 g, 5.14 mmol) was dissolved in DCM (20 ml), (Boc)20 (2.24 g, 10.28 mmol), triethylamine (1.66 g, 16.45 mmol) and DMAP (0.13 g, 1.02 mmol) were added, stirred at room temperature for 16 hours. LCMS monitoring reaction completion. With saturated NaCl aqueous solution (60 ml) and DCM (50 ml) extraction, the organic phase was combined and dried over anhydrous Na2S04, concentrated under reduced pressure, purified by normal phase column chromatography to obtain compound E29-3 (0.54 g, 33.8%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + = 315.2.

[0552] Third Step: Synthesis of compound E29-4

[0553] Compound E29-3 (500 mg, 1.59 mmol) was dissolved in DMF (15 ml), TEA (322 mg, 3.18 mmol), triethylsilane (462 mg, 3.98 mmol), [1,1-bis(diphenylphosphino)ferrocene] dichloropalladium (116.6 mg, 0.19 mmol) were added in turn at room temperature, CO was replaced, and the system was transferred to 100°C and stirred for 16 hours. LCMS monitoring reaction completion. The reaction liquid was extracted with EA (40 ml*2) and water (60 ml), the organic phase was combined and dried over anhydrous sodium sulfate, concentrated and purified by normal phase column chromatography to obtain compound E29-4 (25 mg, 6.1%) as a yellow oil. LC-MS (ESI): m / z found [M+H] + = 264.0.

[0554] Fourth Step: Synthesis of compound E29-5

[0555] Compound E29-4 (25 mg, 0.095 mmol) was dissolved in nitromethane (5 ml), ammonium acetate (29.3 mg, 0.38 mmol) was added, and the reaction was refluxed overnight. LCMS monitoring reaction completion. The reaction liquid was concentrated under reduced pressure, concentrated and purified by normal phase to obtain compound E29-5 (25 mg, 86.2%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + = 307.3.

[0556] Step 5: Synthesis of compound E29-6

[0557] Compound E29-5 (25 mg, 0.082 mmol) was dissolved in DCM (3 ml), hydrochloric acid-dioxane (1.5 mL) was added, and the reaction was stirred at room temperature for 2 h. The reaction was monitored by LCMS to be completed. The reaction solution was concentrated under reduced pressure, and the concentrated solution was purified by normal phase to give compound E29-5 (15 mg, 89.3%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + = 207.3.

[0558] Step 6: Synthesis of compound E29-7

[0559] Compound E29-6 (15 mg, 0.073 mmol) was dissolved in THF (20 ml), and lithium aluminum hydride (0.43 ml, 2.5 mol in THF) was added, and the reaction was stirred at room temperature for 2 h. The reaction was monitored by LCMS to be completed. H2O (2.5 ml), 15% NaOH aqueous solution (2.5 ml), and H2O (7.5 ml) were added dropwise successively under ice water bath cooling, the ice water bath was removed, and the reaction was stirred at room temperature for 1 h. The reaction solution was filtered through diatomite, and the filtrate was concentrated under reduced pressure. The concentrated solution was purified by HPLC to give compound E29-7 (10 mg, 77.2%) as a yellow oil. LC-MS (ESI): m / z found [M+H] + = 179.2.

[0560] Step 7: Synthesis of compound E29

[0561] Compound E29-7 (10 mg, 0.064 mmol) was dissolved in AcOH (2 ml), and M24 (10 mg, 0.016 mmol) was added, and the reaction was stirred at room temperature for 16 h. The reaction was monitored by LCMS to be completed. The reaction solution was concentrated, extracted with EA (5 ml*2) and saturated sodium bicarbonate aqueous solution (10 ml), and the organic phase was combined, dried over anhydrous sodium sulfate, and concentrated. The concentrated solution was purified by HPLC to give compound E29 (7.48 mg, 59.5%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 782.8.

[0562] Example 23: Synthesis of compound E31

[0563] Step 1: Synthesis of compound E31-2

[0564] E31-1 (1 g, 4.67 mmol) was dissolved in ethanol (40 ml), triethylamine (1.41 g, 14.02 mmol) and Pd(dppf)Cl2(342 mg, 0.467 mmol) were added, CO was replaced for three times, heated to 100 °C and stirred for 2 hours. The reaction was monitored by LCMS to be completed. The reaction was cooled and poured into water (200 ml), extracted with EA (100 ml*2), the organic phase was combined, dried and purified by normal phase column chromatography. Compound E31-2 (460 mg, 47.5%) was obtained as a yellow solid after concentration. LC-MS (ESI): m / z found [M+H]=207.20. +

[0565] Second step: synthesis of compound E31-3

[0566] DMF (4.5 ml) was cooled in ice bath and replaced with nitrogen for three times. Phosphorus oxychloride (3.4 g, 22.11 mmol) was added and stirred at room temperature for 15 minutes. Compound E31-2 (460 mg, 2.21 mmol) was dissolved in DMF (1.5 ml) and added to the reaction system. The reaction was stirred at room temperature for 1 hour and then heated to 40 °C for 2 hours. The reaction was monitored by LCMS to be completed. The reaction was cooled and poured into water (50 ml), the pH was adjusted to basic with saturated sodium hydroxide solution, filtered and dried to obtain compound E31-3 (400 mg, 76.9%) as a white solid. LC-MS (ESI): m / z found [M+H]=235.21. +

[0567] Third step: synthesis of compound E31-4

[0568] Compound E31-3 (400 mg, 2.47 mmol) was dissolved in nitromethane (8 ml), ammonium acetate (346 mg, 4.49 mmol) was added, and nitrogen was replaced for three times. The reaction was stirred at 102 °C for 1.5 hours. The reaction was monitored by LCMS to be completed. The reaction was poured into water (30 ml), extracted with EA (20 ml*2), the organic phase was combined, dried, concentrated and purified by normal phase column chromatography. Compound E31-4 (222 mg, 32.3%) was obtained as a yellow solid after concentration. LC-MS (ESI): m / z found [M+H]=278.24. +

[0569] Fourth step: synthesis of compound E31-5

[0570] ​​​Compound E31-4 (222 mg, 0.798 mmol) was dissolved in DMF (2 mL) and methanol (2 ml), sodium borohydride (60.7 mg, 1.60 mmol) was added in three portions under ice bath, and the reaction was stirred at room temperature for 1.5 hours. LCMS monitoring showed the reaction was completed. The reaction solution was poured into water (10 ml), and the pH was adjusted to 5-6 by adding 1M HC1, then extracted with EA (6 ml*2), the organic phase was combined, dried and concentrated, and purified by normal phase column chromatography to obtain E31-5 (100 mg, 44.7%) as a yellow solid. LC-MS (ESI): m / z found [M+H] = 280.26. + = 280.26.

[0571] Step 5: Synthesis of compound E31-6

[0572] Compound E31-5 (90 mg, 0.391 mmol) was dissolved in THF (2 ml), cooled in an ice water bath, and replaced with nitrogen three times, then lithium aluminum hydride (2.5M, 1.56 ml) was added, and the reaction was stirred at room temperature for 16 hours. LCMS monitoring showed the reaction was completed. The reaction solution was purified by HPLC preparation, and freeze-dried to obtain compound E31-6 (16 mg, 19.6%) as a white solid. LC-MS (ESI): m / z found [M+H] = 208.24. + = 208.24.

[0573] Step 6: Synthesis of compound E31

[0574] Compound E31-6 (16 mg, 0.0768 mmol) was dissolved in AcOH (1 ml), and Int.1 (8 mg, 0.0128 mmol) was added, and replaced with nitrogen three times, and the reaction was stirred at room temperature for 16 hours. LCMS monitoring showed the reaction was completed. The reaction solution was purified by HPLC to obtain compound E31 (3.2 mg, 30.8%) as a white solid. LC-MS (ESI): m / z found [M+H] = 811.88. + = 811.88.

[0575] Example 24: Synthesis of compound E32

[0576] Step 1: Synthesis of compound E32-2

[0577] Compound E32-1 (2 g, 10.6 mmol) was dissolved in DMF (80 ml), potassium carbonate (4.4 g, 31.8 mmol) and 1,2-2 bromoethane (2.4 g, 12.7 mmol) were added, and the reaction was carried out at 120 °C for 16 hours. The reaction was monitored by LCMS. The reaction was extracted with saturated NaCl aqueous solution (60 ml) and EA (40 ml), and the organic phase was combined and dried over anhydrous Na2SO4, concentrated under reduced pressure, and purified by normal phase column chromatography to obtain compound E32-2 (0.7 g, 31.8%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + = 215.0.

[0578] Second step: synthesis of compound E32-3

[0579] Compound E32-2 (0.7 g, 3.27 mmol) was dissolved in DCM (20 ml), (Boc)2O (1.43 g, 6.54 mmol), triethylamine (1.06 g, 10.46 mmol) and DMAP (0.079 g, 0.65 mmol) were added, and the reaction was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction was extracted with saturated NaCl aqueous solution (60 ml) and DCM (50 ml), and the organic phase was combined and dried over anhydrous Na2SO4, concentrated under reduced pressure, and purified by normal phase column chromatography to obtain compound E32-3 (0.7 g, 70.0%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + = 315.2.

[0580] Third step: synthesis of compound E32-4

[0581] Compound E32-3 (500 mg, 1.59 mmol) was dissolved in DMF (15 ml), and TEA (322 mg, 3.18 mmol), triethylsilane (462 mg, 3.98 mmol), [1,1-bis(diphenylphosphino)ferrocene]palladium dichloride (116.6 mg, 0.19 mmol) were added in turn at room temperature, CO was replaced, and the system was transferred to 100 °C and stirred for 16 hours. The reaction was monitored by LCMS. The reaction was extracted with EA (40 ml*2) and water (60 ml), and the organic phase was combined and dried over anhydrous sodium sulfate, concentrated, and purified by normal phase column chromatography to obtain compound E32-4 (320 mg, 76.4%) as a yellow oil. LC-MS (ESI): m / z found [M+H] + = 264.0.

[0582] Fourth step: synthesis of compound E32-5

[0583] Compound E32-4 (320 mg, 0.095 mmol) was dissolved in nitromethane (5 ml), ammonium acetate (399.1 mg, 0.38 mmol) was added, and the reaction was refluxed overnight. The reaction was monitored by LCMS. The reaction was concentrated under reduced pressure, and purified by normal phase after concentration to give compound E32-5 (220 mg, 59.1%) as a yellow solid. LC-MS (ESI): m / z found [M+H] = 307.3. +

[0584] Step 5: Synthesis of compound E32-6

[0585] Compound E32-5 (220 mg, 0.718 mmol) was dissolved in DCM (5 ml), hydrochloric acid-dioxane (5 mL) was added, and the reaction was stirred at room temperature for 2 h. The reaction was monitored by LCMS. The reaction was concentrated under reduced pressure, and purified by normal phase after concentration to give compound E32-6 (120 mg, 81.1%) as a yellow solid. LC-MS (ESI): m / z found [M+H] = 207.3. +

[0586] Step 6: Synthesis of compound E32-7

[0587] Compound E32-6 (120 mg, 0.58 mmol) was dissolved in THF (20 ml), lithium aluminum hydride (2.56 ml, 2.5 mol in THF) was added, and the reaction was stirred at room temperature for 2 h. The reaction was monitored by LCMS. H2O (2.5 ml), 15% NaOH aqueous solution (2.5 ml), and H2O (7.5 ml) were added dropwise successively under ice water bath cooling, the ice water bath was removed, and the reaction was stirred at room temperature for 1 h. The reaction was filtered through celite, and the filtrate was concentrated under reduced pressure. The product was purified by HPLC after concentration to give compound E32-7 (100 mg, 96.5%) as a yellow oil. LC-MS (ESI): m / z found [M+H] = 179.2. +

[0588] Step 7: Synthesis of compound E32

[0589] ​​​Compound E32-7 (22 mg, 0.12 mmol) was dissolved in AcOH (2 ml), M24 (19.2 mg, 0.03 mmol) was added, and the mixture was stirred at room temperature for 16 h. The reaction was monitored by LCMS. The reaction solution was concentrated and extracted with EA (5 ml*2) and saturated aqueous sodium bicarbonate solution (10 ml). The organic phase was combined, dried over anhydrous sodium sulfate, concentrated, and purified by HPLC to obtain compound E32 (14 mg, 58.1%) as a white solid. LC-MS (ESI): m / z found [M+H]=782.8. +

[0590] Example 25: Synthesis of compound E33

[0591] Step 1: Synthesis of compound E33

[0592] Compound E26 (2.2 mg, 0.002 mmol) was dissolved in ACN (1.5 ml) and H2O (1 ml), and silver nitrate (8.1 mg, 0.048 mmol) was added. The mixture was stirred at room temperature for 12 h. The reaction was monitored by LCMS. Saturated brine (2.5 ml) and saturated sodium bicarbonate (2.5 ml) were added to the reaction solution, and the mixture was stirred for 15 min. The mixture was extracted with EA (5 ml*2) and water (10 ml). The organic phase was combined, dried over anhydrous sodium sulfate, concentrated, and purified by HPLC to obtain compound E33 (1 mg, 46.1%) as a white solid. LC-MS (ESI): m / z found [M+H]=850.9. +

[0593] Example 26: Synthesis of compound E35

[0594] Step 1: Synthesis of compound E35

[0595] Compound E23 (5 mg, 0.006 mmol) was dissolved in ACN (0.6 ml) and distilled water (0.4 ml), and silver nitrate (19.7 mg, 0.1157 mmol) was added. The mixture was replaced with nitrogen three times and stirred at room temperature for 16 h. The reaction was monitored by LCMS. The reaction solution was purified by HPLC preparation, and compound E35 (2.2 mg, 45.1%) was obtained as a white solid after lyophilization. LC-MS (ESI): m / z found [M+H]=812.94. +

[0596] Example 27: Synthesis of compound E36 ​​​

[0597] Step 1: synthesis of compound E36

[0598] Compound E22 (1.65 mg, 0.0018 mmol) was dissolved in ACN (1.5 ml) and H2O (1 ml), silver nitrate (5.98 mg, 0.035 mmol) was added, stirred at room temperature for 12 hours. LCMS monitoring reaction completion. To the reaction solution was added saturated brine (2.5 ml) and saturated sodium bicarbonate (2.5 ml), stirred for 15 min, the mixture was extracted with EA (5 ml*2) and water (10 ml), the organic phase was combined, dried over anhydrous sodium sulfate, concentrated and purified by HPLC to give compound E36 (0.2 mg, 12.3%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 865.0.

[0599] Example 28: synthesis of compound E37

[0600] Step 1: synthesis of compound E37

[0601] Compound E24 (7.35 mg, 0.008 mmol) was dissolved in ACN (1.5 ml) and H2O (1 ml), silver nitrate (26.7 mg, 0.157 mmol) was added, stirred at room temperature for 12 hours. LCMS monitoring reaction completion. To the reaction solution was added saturated brine (2.5 ml) and saturated sodium bicarbonate (2.5 ml), stirred for 15 min, the mixture was extracted with EA (5 ml*2) and water (10 ml), the organic phase was combined, dried over anhydrous sodium sulfate, concentrated and purified by HPLC to give compound E37 (1.2 mg, 16.5%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 865.0.

[0602] Example 29 & 30: synthesis of compound E38 & E39

[0603] Step 1: synthesis of compound E38-2

[0604] E38-1 (7.5 g, 4.67 mmol) was dissolved in THF (80 ml), BH3.THF (101 ml, 14.02 mmol) was added slowly under ice-bath and N2 protection. The reaction was stirred at room temperature for 2 h. LCMS was used to monitor the reaction completion. The reaction was cooled in ice-bath and poured into ice-water (100 ml). 1 N HCl was added to adjust the pH to acidic. EA (100 ml*2) was used to extract the mixture. The organic phase was combined and dried. The product was purified by normal phase column chromatography. The compound E38-2 (3.3 g, 51.3%) was obtained as a yellow solid after concentration. LC-MS (ESI): m / z found [M+H]=181.14. +

[0605] Second Step: Synthesis of compound E38-4

[0606] E38-2 (3.3 g, 18.333 mmol) was dissolved in ether and cooled in ice-bath. N2 was bubbled for 3 times. Oxalyl chloride (10 ml) was added. The reaction was stirred at room temperature for 15 min and heated to 50 °C for 8 h. The reaction was cooled and filtered. 30 ml of ammonia methanol solution was added. The reaction was stirred at room temperature overnight. LCMS was used to monitor the reaction completion. The product was concentrated and washed with ether for 3 times. The compound E38-4 (4.7 g, 90%) was obtained as a gray solid after drying. LC-MS (ESI): m / z found [M+H]=252.17. +

[0607] Third Step: Synthesis of compound E38-5

[0608] Compound E38-4 (300 mg, 1.190 mmol) was dissolved in THF (6 ml) and cooled in ice-bath. N2 was bubbled for 3 times. Lithium aluminum hydride (2.5 M, 1 ml) was added. The reaction was heated to 50 °C and stirred for 2.5 h. LCMS was used to monitor the reaction completion. The reaction was purified by HPLC. The compound E38-5 (37 mg, 16.0%) was obtained as a white solid after lyophilization. LC-MS (ESI): m / z found [M+H]=194.24. +

[0609] Fourth Step: Synthesis of compound E38

[0610] ​​​Compound E38-5 (37 mg, 0.190 mmol) was dissolved in AcOH (1 ml), added Int. 1 (35 mg, 0.0563 mmol), replaced with nitrogen for three times, stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction was purified by HPLC to give compound E38 (10 mg, 30.8%) as a white solid. LC-MS (ESI): m / z found [M+H]=797.27. +

[0611] Fifth step: synthesis of compound E39

[0612] Compound E38 (10 mg, 0.0125 mmol) was dissolved in ACN:H2O (3 ml), added AgNO3 (42 mg, 0.250 mmol), replaced with nitrogen for three times, stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction was purified by HPLC to give compound E39 (5 mg, 50%) as a white solid. LC-MS (ESI): m / z found [M+H]=788.27. +

[0613] Example 31: synthesis of compound E40

[0614] First step: synthesis of compound E40

[0615] Compound E39 (6 mg, 0.0076 mmol), glycolic acid (0.75 mg, 0.0098 mmol) was dissolved in DMF (2 ml), added EDCI (1.9 mg, 0.0098 mmol), HOBT (0.3 mg, 0.0022 mmol), TEA (2 ul), reacted at room temperature for 2 hours. Compound E40 (0.5 mg, 7.7%) was obtained as a white solid after HPLC purification. LC-MS (ESI): m / z found [M+H]=846.27. +

[0616] Example 32: synthesis of compound E41

[0617] First step: synthesis of compound E41-2

[0618] ​​​Compound E41-1a (450 mg, 1.42 mmol) was dissolved in 1,4-dioxane (9 ml), TEA (1.35 ml), E23-4A (99.37 mg, 1.42 mmol), cuprous iodide (81.13 mg, 0.426 mmol) and dichlorobis(triphenylphosphine)palladium (99.68 mg, 0.142 mmol) were added, and the mixture was stirred at room temperature for 3 hours under nitrogen. The reaction was monitored by LCMS. The reaction was poured into water (30 ml), and extracted with EA (15 ml*2). The organic phase was combined and dried, and concentrated. Purification was performed by normal phase column chromatography to give compound E41-2 (260 mg, 70.9%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=258.28. +

[0619] Second Step: Synthesis of compound E41-3

[0620] Compound E41-2 (260 mg, 1.006 mmol) was dissolved in MeOH (10 ml), and Pd-C (780 mg, 300% wt) was added. The mixture was stirred at room temperature for 16 hours under hydrogen. The reaction was monitored by LCMS. The reaction was filtered and concentrated to give compound E41-3 (130 mg, 55.7%) as a yellow oily solid. LC-MS (ESI): m / z found [M+H]=232.33. +

[0621] Third Step: Synthesis of compound E41

[0622] Compound E41-3 (26 mg, 0.11 mmol) was dissolved in AcOH (1 ml), and Int.1 (15 mg, 0.024 mmol) was added. The mixture was stirred at room temperature for 16 hours under nitrogen. The reaction was monitored by LCMS. The reaction was purified by HPLC to give compound E41 (3.7 mg, 18.4%) as a white solid. LC-MS (ESI): m / z found [M+H]=835.97. +

[0623] Example 33: Synthesis of compound E42

[0624] First Step: Synthesis of compound E42

[0625] ​​​Compound E41 (7.8 mg, 0.0093 mmol) was dissolved with ACN (0.6 ml) and distilled water (0.4 ml), silver nitrate (30 mg, 0.177 mmol) was added, replaced with nitrogen for three times, stirred at room temperature for 16 h. The reaction was monitored by LCMS. The reaction was purified by HPLC to give compound E42 (3.74 mg, 48.1%) as a white solid. LC-MS (ESI): m / z found [M+H]=835.97. +

[0626] Example 34: Synthesis of compound E44

[0627] Step 1: Synthesis of compound E44

[0628] Compound E29 (11.6 mg, 0.015 mmol) was dissolved in ACN (1.5 ml) and H2O (1 ml), silver nitrate (47.9 mg, 0.29 mmol) was added, stirred at room temperature for 12 hours. The reaction was monitored by LCMS. To the reaction was added saturated brine (2.5 ml) and saturated sodium bicarbonate (2.5 ml), stirred for 15 min, the mixture was extracted with EA (5 ml*2) and water (10 ml), the organic phase was combined and dried over anhydrous sodium sulfate, concentrated and purified by HPLC to give compound E44 (5.5 mg, 48.2%) as a white solid. LC-MS (ESI): m / z found [M+H]=772.8. +

[0629] Example 35: Synthesis of compound E45

[0630] Step 1: Synthesis of compound E45

[0631] Compound E31 (50 mg, 0.0627 mmol) was dissolved in ACN:H2O (3 ml), AgNO3 (211 mg, 1.2588 mmol) was added, replaced with nitrogen for three times, stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction was purified by HPLC to give compound E45 (20 mg, 38.1%) as a white solid. LC-MS (ESI): m / z found [M+H]=802.87. +

[0632] Example 36: Synthesis of compound E46

[0633] Step 1: Synthesis of compound E46-2 ​​​

[0634] Compound E46-1 (500 mg, 1.72 mmol) was dissolved in DCM (10 ml), and Dess-Martin Oxidizing Agent (1.093 g, 2.58 mmol) was added, and stirred at room temperature for 2 hours. LCMS monitoring showed that the reaction was completed. The reaction solution was poured into water (50 ml), and extracted with EA (30 ml*2), and the organic phase was combined and dried over anhydrous sodium sulfate, and purified by normal phase column chromatography, and concentrated to give compound E46-2 (276 mg, 55.6%) as a brown oil. LC-MS (ESI): m / z found [M+H] + = 288.35.

[0635] Second step: synthesis of compound E46-3

[0636] Compound E46-2 (200 mg, 0.693 mmol) was dissolved in DCM (4 ml) and MeOH (2 ml), and methylamine hydrochloride (186 mg, 2.77 mmol) and NaBH3CN (131 mg, 2.08 mmol) were added, and replaced with nitrogen three times, and stirred at room temperature for 16 hours. LCMS monitoring showed that the reaction was completed. The reaction solution was concentrated and dissolved in methanol, and purified by reversed phase column chromatography, and concentrated to give compound E46-3 (120 mg, 39.5%) as a brownish yellow solid. LC-MS (ESI): m / z found [M+H] + = 303.41.

[0637] Third step: synthesis of compound E46-4

[0638] Compound E46-3 (100 mg, 0.33 mmol) was dissolved in DCM (10 mL), and trifluoroacetic acid (376.2 mg, 3.3 mmol) was added, and stirred at room temperature for 1.5 hours. LCMS monitoring showed that the reaction was completed. The reaction solution was concentrated and dissolved in methanol, and purified by HPLC preparation, and freeze-dried to give E46-4 (26 mg, 38.7%) as a colorless transparent oil. LC-MS (ESI): m / z found [M+H] + = 203.29.

[0639] Fourth step: synthesis of compound E46

[0640] Compound E46-4 (10 mg, 0.0491 mmol) was dissolved in AcOH (1 ml), added Int. 1 (5 mg, 0.008 mmol), replaced with nitrogen for three times, stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction was purified by HPLC to give compound E46 (1 mg, 15.5%) as a white solid. LC-MS (ESI): m / z found [M+H]= 806.94. + = 806.94.

[0641] Example 37: Synthesis of compound E47

[0642] First Step: Synthesis of compound E47-2

[0643] Compound E47-1 (30 mg, 0.0987 mmol) was dissolved in DMF (1 ml), added glycolic acid (15.6 mg, 0.197 mmol), HOBt (8 mg, 0.0592 mmol) and EDCI (49.3 mg, 0.257 mmol), stirred at room temperature for 5 min, added TEA (30 mg, 0.296 mmol), stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction was purified by reverse phase column chromatography, concentrated to give compound E47-2 (17.5 mg, 49.1%) as a light yellow solid. LC-MS (ESI): m / z found [M+H]= 361.44. + = 361.44.

[0644] Second Step: Synthesis of compound E47-3

[0645] Compound E47-2 (17.5 mg, 0.048 mmol) was dissolved in DCM (0.6 mL), added trifluoroacetic acid (55.1 mg mg, 0.483 mmol), reacted at room temperature for 1.5 hours. The reaction was monitored by LCMS. The reaction was concentrated, dissolved in methanol, purified by HPLC preparation, freeze-dried to give E47-3 (5 mg, 39.7%) as a colorless transparent oil. LC-MS (ESI): m / z found [M+H]= 261.33. + = 261.33.

[0646] Third Step: Synthesis of compound E47

[0647] Compound E47-3 (5 mg, 0.019 mmol) was dissolved in AcOH (0.5 ml), added Int. 1 (4 mg, 0.006 mmol), replaced with nitrogen for three times, stirred at room temperature for 16 hours. The reaction was monitored to completion by LCMS. The reaction was purified by HPLC to give compound E47 (1.4 mg, 8.5%) as a white solid. LC-MS (ESI): m / z found [M+H]=864.97. +

[0648] Example 38: Synthesis of compound E48

[0649] First Step: Synthesis of compound E48-2

[0650] Compound E48-1 (0.5 g, 2.6 mmol) was dissolved in THF (6 ml), cooled with ice water bath, replaced with nitrogen for three times, added lithium hydroxide monohydrate (550 mg, 13.0 mmol), stirred at room temperature for 2.5 h. The reaction was monitored to completion by LCMS. Purified by normal phase column chromatography with 55% PE in EA to give product, concentrated to give E48-2 (0.4 g, 85.5%) as a white solid. LC-MS (ESI): m / z found [M+H]=178.2. +

[0651] Second Step: Synthesis of compound E48-3

[0652] Compound E48-2 (400 mg, 2.2 mmol) and HATU (1.16 g, 1.0 mmol) were dissolved in DMF (5 ml), added N, O-dimethylhydroxylamine (270 mg, 4.4 mmol) and DIEA (711 mg, 5.5 mmol), replaced with nitrogen for three times, stirred at room temperature for 1 h. The reaction was monitored to completion by LCMS. The reaction was purified by reverse phase column chromatography with 35% ACN in H2O to give compound E48-3 (380 mg, 76.5%) as a white solid. LC-MS (ESI): m / z found [M+H]=221.3 +

[0653] Third Step: Synthesis of compound E48-4

[0654] ​​​Compound E48-3 (380 mg, 1.8 mmol) was dissolved in THF (6 ml), cooled at -40 °C, replaced by nitrogen for three times, added lithium hydroxide (1.2 ml) slowly, stirred at -40 °C for 1 h. LCMS monitored the reaction was completed. Added saturated ammonium chloride to quench, washed the aqueous phase by ethyl acetate for three times, collected the oil phase, dried by anhydrous sodium sulfate, concentrated to get E48-4 (200 mg, crude) as yellow liquid. LC-MS (ESI): m / z found [M+H] + = 162.5.

[0655] Fourth step: synthesis of compound E48-5

[0656] E48-4 (200 mg, 1.23 mmol) was dissolved in CH3NO2 (80 ml), added NH4OAc (250 mg, 3.2 mmol) slowly under nitrogen protection, stirred at 102 °C for 2 h. LCMS monitored the reaction was completed. The reaction solution was cooled in ice bath, poured into ice water (100 ml), stirred in ice water bath for half an hour, yellow solid precipitated, filtered and concentrated to get compound E48-5 (120 mg, 47.2%) as yellow solid. LC-MS (ESI): m / z found [M+H] + = 204.5.

[0657] Fifth step: synthesis of compound E48-6

[0658] Compound E48-5 (120 mg, 0.58 mmol) was dissolved in THF (3 ml), replaced by nitrogen for three times in ice bath, added LiAlH4 (0.5 ml) slowly, stirred at room temperature for 12 h. LCMS monitored the reaction was completed. The reaction solution was quenched by saturated ammonium chloride, washed the aqueous phase by ethyl acetate for three times, collected the oil phase, dried by anhydrous sodium sulfate, concentrated, purified by Pre-HPLC to get compound E48-6 (57 mg, 36.8%) as yellow liquid. LC-MS (ESI): m / z found [M+H] + = 176.6.

[0659] Sixth step: synthesis of compound E48

[0660] Compound E48-6 (57 mg, 0.32 mmol) was dissolved in AcOH (1 ml), added M24 (C 44 H 40 Cl3N5O 11S, 90 mg, 0.064 mmol), nitrogen was replaced for three times, stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction was purified by HPLC to give compound E48 (20 mg, 29%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 779.6.

[0661] Example 39: Synthesis of compound E49

[0662] First step: Synthesis of compound E49-2

[0663] Compound E49-1 (250 mg, 1.01 mmol) was dissolved in nitromethane (5 ml), ammonium acetate (205.5 mg, 2.67 mmol) was added, nitrogen was replaced for three times, stirred at 102 °C for 1.5 hours. The reaction was monitored by LCMS. The reaction was poured into water (30 ml), extracted with EA (20 ml*2), the organic phase was combined and dried, concentrated and purified by normal phase column chromatography, concentrated to give compound E49-2 (210 mg, 71.7%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + = 290.32.

[0664] Second step: Synthesis of compound E49-3

[0665] Compound E49-2 (210 mg, 0.724 mmol) was dissolved in DCM (5 mL), trifluoroacetic acid (1 ml) was added, and the reaction was stirred at room temperature for 1.5 hours. The reaction was monitored by LCMS. The reaction was added to saturated sodium bicarbonate solution (20 ml), extracted with EA (15 ml) twice, the organic phase was combined and dried with anhydrous sodium sulfate, concentrated to give E49-3 (120 mg, 87.2%) as a light yellow solid. LC-MS (ESI): m / z found [M+H] + = 190.20.

[0666] Third step: Synthesis of compound E49-4

[0667] Compound E49-3 (120 mg, 0.624 mmol) was dissolved in methanol (6 ml) and DMF (6 ml), cooled in ice water bath, replaced with nitrogen for three times, sodium borohydride (48 mg, 1.248 ml) was added, stirred at room temperature for 2 hours. LCMS monitored the completion of the reaction. The reaction solution was purified by HPLC, and Compound E49-4 (60 mg, 50%) was obtained as a white solid after lyophilization. LC-MS (ESI): m / z found [M+H]=192.22. +

[0668] Fourth Step: Synthesis of Compound E49-5

[0669] Compound E49-4 (60 mg, 0.312 mmol) was dissolved in methanol (1.8 ml), palladium on carbon (60 mg) was added, replaced with hydrogen for three times, stirred at room temperature for 16 hours. LCMS monitored the completion of the reaction. The reaction solution was filtered, and the filtrate was concentrated to obtain Compound E49-5 (40 mg, 79.1%) as a white solid. LC-MS (ESI): m / z found [M+H]=162.24. +

[0670] Fifth Step: Synthesis of Compound E49

[0671] Compound E49-5 (6 mg, 0.032 mmol) was dissolved in AcOH (0.5 ml), Int.1 (5 mg, 0.008 mmol) was added, replaced with nitrogen for three times, stirred at room temperature for 16 hours. LCMS monitored the completion of the reaction. The reaction solution was purified by HPLC to obtain Compound E49 (0.7 mg, 11.4%) as a white solid. LC-MS (ESI): m / z found [M+H]=765.88. +

[0672] Example 40: Synthesis of Compound E50

[0673] First Step: Synthesis of Compound E50

[0674] E48 (20 mg, 0.025 mmol) was dissolved in acetonitrile: water 3:2, AgNO3 (218 mg, 0.46 mmol) was added, replaced with nitrogen for three times, stirred at room temperature for 16 hours. LCMS monitored the completion of the reaction. The reaction solution was purified by HPLC to obtain Compound E50 (10 mg, 51%) as a white solid. LC-MS (ESI): m / z found [M+H]=770.0. + ​​​​

[0675] Example 41: Synthesis of compound E51:

[0676] First Step: Synthesis of compound E51-2

[0677] Compound E51-1 (2 g, 11.9 mmol) was dissolved in DMF (80 ml), potassium carbonate (4.9 g, 35.9 mmol) and 1,2-2 bromoethane (2.7 g, 14.2 mmol) were added, and the reaction was carried out at 120 °C for 16 hours. The reaction was monitored by LCMS. The reaction was extracted with saturated NaCl aqueous solution (60 ml) and EA (40 ml), and the organic phase was combined and dried over anhydrous Na2SO4, concentrated under reduced pressure, and purified by normal phase column to obtain compound E51-2 (0.8 g, 34.8%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + = 194.2.

[0678] Second Step: Synthesis of compound E51-3

[0679] Compound E51-2 (800 mg, 4.14 mmol) was dissolved in ETOH (5 ml) and water (5 ml), and lithium hydroxide (64.62 mg, 8.29 mmol) was added, and the reaction was stirred at room temperature for 4 hours. The reaction was monitored by TLC. The reaction was adjusted to PH with dilute hydrochloric acid, extracted with EA (10 ml*2), and the organic phase was combined and dried over anhydrous sodium sulfate, concentrated to obtain compound E51-3 (600 mg, 80.8%) as a colorless oil. LC-MS (ESI): m / z found [M+H] + = 180.0.

[0680] Third Step: Synthesis of compound E51-4

[0681] Compound E51-3 (600 mg, 3.35 mmol) was dissolved in DMF (6 ml), N, O-dimethylhydroxylamine (361.3 mg, 3.68 mmol), HATU (1.91 g, 5.02 mmol), and DIEA (864.8 mg, 6.70 mmol) were added, and the reaction was stirred at room temperature for 1 hour. The reaction was monitored by LCMS. EA (10 ml) was added to the reaction, and saturated sodium bicarbonate (20 ml) was washed, and the organic phase was dried, filtered, and concentrated, and purified by HPLC to obtain compound E51-4 (230 mg, 30.9%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + = 223.2.

[0682] Step 4: Synthesis of compound E51-5

[0683] Compound E51-4 (230 mg, 1.036 mmol) was dissolved in THF (15 mL), and lithium aluminum hydride (0.8 mL, 2.5 mol in THF) was added at -45 °C. The reaction was allowed to proceed for 2 hours. The reaction was monitored by LC-MS until completion. Under ice-water bath cooling, H2O (2.5 mL), 15% NaOH aqueous solution (2.5 mL), and H2O (7.5 mL) were added dropwise. The ice-water bath was removed, and the mixture was stirred at room temperature for 1 hour. The mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure. After concentration, it was purified by HPLC to obtain compound E51-5 (70 mg, 41.6%) as a yellow oil. LC-MS (ESI): m / z found [M+H] + =164.1.

[0684] Step 5: Synthesis of compound E51-6

[0685] Compound E51-5 (70 mg, 0.429 mmol) was dissolved in nitromethane (5 mL), and ammonium acetate (132.2 mg, 1.71 mmol) was added. The mixture was refluxed overnight. The reaction was monitored by LC-MS to ensure completion. The reaction solution was concentrated under reduced pressure and then purified in normal phase to give compound E51-6 (50 mg, 88.5%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + =207.2.

[0686] Step 6: Synthesis of compound E51-7

[0687] Compound E51-6 (50 mg, 0.24 mmol) was dissolved in THF (10 mL), and lithium aluminum hydride (1.06 mL, 2.5 mol in THF) was added. The reaction was carried out at room temperature for 2 hours. The reaction was monitored by LC-MS. H2O (2.5 mL), 15% NaOH aqueous solution (2.5 mL), and H2O (7.5 mL) were added dropwise under ice-water bath cooling. The ice-water bath was removed, and the mixture was stirred at room temperature for 1 hour. The mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure. After concentration, it was purified by HPLC to obtain compound E51-7 (15 mg, 34.5%) as a yellow oil. LC-MS (ESI): m / z found [M+H] + =179.2.

[0688] Step 7: Synthesis of compound E51

[0689] Compound E51-7 (15 mg, 0.084 mmol) was dissolved in AcOH (2 ml), M24 (13.1 mg, 0.021 mmol) was added, and the mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction was concentrated and extracted with EA (5 ml*2) and saturated aqueous NaHC03solution (10 ml). The organic phase was combined and dried over anhydrous Na2S04. After concentration, HPLC purification was performed to obtain compound E51 (0.8 mg, 1.21%) as a white solid. LC-MS (ESI): m / z found [M+H]+= 782.8.

[0690] Example 42: Synthesis of compound E52

[0691] First step: Synthesis of compound E52-2

[0692] Compound E52-1 (0.4 g, 2.45 mmol) was dissolved in DCM (20 ml), DBU (C9H 16 N2, 391.6 mg, 2.57 mmol) and SM2 (C 12 H 14 N4O2S, 561.2 mg, 2.69 mmol) were added, and the mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. Extraction was performed with saturated aqueous NaCl solution (60 ml) and EA (40 ml). The organic phase was combined and dried over anhydrous Na2S04. After concentration under reduced pressure, normal phase column purification was performed to obtain compound E52-2 (0.2 g, 24.4%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + = 335.3.

[0693] Second step: Synthesis of compound E52-3

[0694] Compound E52-2 (200 mg, 0.59 mmol) was dissolved in MeOH (26.6 mL), and Pd / C (20 mg, 60% wt) was added. Hydrogen replacement was performed, and the mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. After filtration and concentration, crude E52-3 (80 mg) was obtained. LC-MS (ESI): m / z found [M+H] + = 337.3.

[0695] Third step: Synthesis of 3-butyl(l-(3,4-dihydro-2H-ben[b][l,4]oxazin-8-yl)-3-oxopropan-2-yl) carbonate

[0696] Compound E52-3 (80 mg, 0.23 mmol) was dissolved in THF (15 ml), lithium borohydride (0.3 ml, 2 mol in THF) was added at room temperature, and the mixture was reacted at 60 °C for 16 hours. The reaction was monitored by LCMS. H2O (2.5 ml), 15% NaOH aqueous solution (2.5 ml), and H2O (7.5 ml) were added dropwise successively under ice water bath cooling, the ice water bath was removed, and the mixture was filtered through diatomite. The filtrate was concentrated under reduced pressure, and the concentrate was purified by HPLC to obtain compound E52-4 (50 mg, 73.3%) in the form of yellow oil. LC-MS (ESI): m / z found [M+H]+ = 309.3. +

[0697] Fourth step: synthesis of compound E52-5

[0698] Compound E52-4 (50 mg, 0.16 mmol) was dissolved in DCM (3 ml), and hydrochloric acid-dioxane (1.5 mL) was added, and the mixture was reacted at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction solution was concentrated under reduced pressure, and the concentrate was purified by normal phase to obtain compound E52-5 (30 mg, 88.8%) in the form of yellow solid. LC-MS (ESI): m / z found [M+H]+ = 209.2. +

[0699] Fifth step: synthesis of compound E52

[0700] Compound E52-5 (20 mg, 0.096 mmol) was dissolved in AcOH (2 ml), and M24 (14.9 mg, 0.024 mmol) was added, and the mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction solution was concentrated, extracted with EA (5 ml*2) and saturated aqueous sodium bicarbonate solution (10 ml), and the organic phase was combined, dried over anhydrous sodium sulfate, and concentrated. The concentrate was purified by HPLC to obtain compound E52 (2.4 mg, 12.3%) in the form of white solid. LC-MS (ESI): m / z found [M+H]+ = 812.9.

[0701] Example 43: synthesis of compound E53

[0702] First step: synthesis of compound E53-2

[0703] ​​Compound E52-4 (0.2 g, 0.64 mmol) was dissolved in THF (10 ml), triphenylphosphine (510.4 mg, 1.92 mmol), phthalimide (143.2 mg, 0.96 mmol) and DEAD (282.5 mg, 1.92 mmol) were added, and the reaction was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction was extracted with saturated aqueous NaCl solution (60 ml) and EA (40 ml), and the organic phase was combined and dried over anhydrous Na2SO4. After being concentrated under reduced pressure, the product was purified by normal phase column to give compound E53-2 (0.23 g, 81.0%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=438.5. +

[0704] Second Step: Synthesis of compound E53-3

[0705] Compound E53-2 (80 mg, 0.18 mmol) was dissolved in DCM (20 ml), (Boc)2O (79.8 mg, 0.36 mmol), triethylamine (55.4 mg, 0.54 mmol) and DMAP (4.46 mg, 0.018 mmol) were added, and the reaction was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction was extracted with saturated aqueous NaCl solution (10 ml) and DCM (8 ml), and the organic phase was combined and dried over anhydrous Na2SO4. After being concentrated under reduced pressure, the product was purified by normal phase column to give compound E53-3 (90 mg, 91.5%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=538.6. +

[0706] Third Step: Synthesis of compound E53-4

[0707] Compound E53-3 (240 mg, 0.44 mmol) was dissolved in ETOH (15 ml), hydrazine hydrate (186 mg) was added at room temperature, and the reaction was stirred at 60 °C for 3 hours. The reaction was monitored by LCMS. The filtrate was concentrated under reduced pressure, and the product was purified by HPLC to give compound E53-4 (150 mg, 82.8%) as a yellow oil. LC-MS (ESI): m / z found [M+H]=408.5. +

[0708] Fourth Step: Synthesis of compound E53-5

[0709] ​​​Compound E53-4 (150 mg, 0.36 mmol) was dissolved in DCM (20 ml), and Alloc-Cl (48.9 mg, 0.36 mmol) and DIEA (57 mg, 0.36 mmol) were added, and stirred at room temperature for 5 hours. The reaction was monitored by LCMS. The reaction was extracted with saturated NaCl aqueous solution (10 ml) and DCM (8 ml), and the organic phase was combined and dried over anhydrous Na2SO4, and concentrated under reduced pressure, and purified by normal phase column chromatography to obtain compound E53-5 (130 mg, 71.8%) as a yellow solid. LC-MS (ESI): m / z found [M+H]+ = 492.5. +

[0710] Step 5: Synthesis of compound E53-6

[0711] Compound E53-5 (100 mg, 0.082 mmol) was dissolved in DCM (3 ml), and hydrochloric acid-dioxane (1.5 mL) was added, and reacted at room temperature for 2 h. The reaction was monitored by LCMS. The reaction solution was concentrated under reduced pressure, and purified by normal phase after concentration to obtain compound E53-6 (70 mg) as a yellow solid. LC-MS (ESI): m / z found [M+H]+ = 292.3. +

[0712] Step 6: Synthesis of compound 53-7

[0713] Compound E53-6 (70 mg, 0.24 mmol) was dissolved in AcOH (2 ml), and M24 (37.4 mg, 0.06 mmol) was added, and stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction solution was concentrated and extracted with EA (5 ml*2) and saturated sodium bicarbonate aqueous solution (10 ml), and the organic phase was combined and dried over anhydrous sodium sulfate, and concentrated and purified by HPLC to obtain compound 53-7 (15 mg, 28.2%) as a white solid. LC-MS (ESI): m / z found [M+H]+ = 896.0.

[0714] Step 7: Synthesis of compound E53

[0715] ​​Compound E53-7 (10 mg, 0.01 mmol) was dissolved in DCM (2 ml), Pd(pph3)2Cl2 (1.2 mg, 0.001 mmol), acetic acid (6.7 mg, 0.11 mmol) and Bu3SnH (19.5 mg, 0.06 mmol) were added, stirred at room temperature for 2 hours. LCMS monitoring reaction completion. The reaction was concentrated and extracted with EA (3 ml*2) and saturated aqueous sodium bicarbonate solution (5 ml), the organic phase was combined and dried over anhydrous sodium sulfate, concentrated and purified by HPLC to obtain compound E53 (3 mg, 33.1%) as a white solid. LC-MS (ESI): m / z found [M+H]=811.9. +

[0716] Synthesis of Drug Linker Conjugate (Linker-Payload (L-P))

[0717] The structure of L-P is as follows:

[0718] The structure of the common intermediate Linker, 4-2 involved in the examples is as follows, and its synthesis steps refer to the synthesis steps in the patent WO2024230752 A1;

[0719] The structure of 4-2 is as follows, and its synthesis steps refer to the synthesis steps of “107-8” in WO2024230752 A1:

[0720] The structure and synthesis steps of L2 are as follows:

[0721] Referring to the synthesis steps of “107-8” in WO2024230752 A1, the intermediate 83-11 in compound 107-8 (compound 4-2) is replaced by Other steps are consistent with the synthesis of 107-8, and finally compound L2 is obtained.

[0722] The structure and synthesis steps of L44 are as follows:

[0723] First step: synthesis of intermediate 1-3:

[0724] ​​Dissolve 1-1 (300 mg, 1.030 mmol) in DMF (5 mL), add HOSU (178 mg, 1.545 mmol) and EDCI (296 mg, 1.545 mmol), react at room temperature for 2 h, then add 1-2 (654 mg, 1.545 mmol) and TEA (157 μL, 1.133 mmol) successively, react at room temperature overnight. Monitor the reaction completion by LC-MS, purify by reverse phase column chromatography, and concentrate under reduced pressure to obtain a white solid (363 mg, 51%). LC-MS (ESI): m / z found [M-1] - = 695.38.

[0725] Second Step: Synthesis of L44

[0726] Dissolve 1-3 (120 mg, 0.173 mmol) in THF (3 mL) and EtOH (9 mL), add CuSO4·5H2O (36 mg, 0.144 mmol) and ascorbic acid (25 mg, 0.144 mmol). React at room temperature for 2 h under nitrogen protection. Monitor the reaction completion by LC-MS, purify by reverse phase column chromatography, and freeze-dry to obtain a yellow solid product (143 mg, 55%). LC-MS (ESI): m / z found [M / 2+1] + = 746.10.

[0727] Example 44: Synthesis of compound L44-E1’

[0728] Synthesis of compound L44-E1’

[0729] Dissolve compound L44 (8.9 mg, 0.006 mmol) in DMF (0.5 mL), add DIEA (1.56 mg, 0.012 mmol) and HATU (2.54 mg, 0.0072 mmol) under ice-water bath, and then raise the system to room temperature and react for 30 min, add compound E1’ (5 mg, 0.006 mmol) to the system, and continue to react for 2 h. Monitor the reaction completion by LCMS, pour the reaction liquid into water, extract with EA to obtain an organic phase, concentrate, and purify by prep-HPLC to obtain compound L44-E1 (1.2 mg) as a light yellow solid. LC-MS (ESI): m / z found [1 / 2M+H] + = 1155.7.

[0730] Example 45: Synthesis of compound L42-E2

[0731] First Step: Synthesis of compound E2-5

[0732] To a solution of 4-((S)-2-((S)-2-(2-azidoacetamido)-3-methylbutanamido)-5- ureidopentanoyl)benzyl(4-nitrophenyl) carbonate (3.75 mg, 0.006 mmol) in DMF (1 ml) was added HOBT (0.89 mg, 0.0066 mmol), DIEA (1.55 mg, 0.012 mmol) and compound E2 (5 mg, 0.006 mmol) and stirred at room temperature for 2 hours. The reaction was monitored to completion by LCMS. The reaction was purified by reverse phase column chromatography to give compound E2-5 (3.2 mg, 40.2%) as a white solid. LC-MS (ESI): m / z found [M+H] = 1324.5. +

[0733] Second Step: Synthesis of compound L42-E2

[0734] To a solution of 1-(3,5-difluoro-4-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)phenyl)-3,4- bis((4-(morpholine-4-carbonyl)phenyl)sulfanyl)-1H-pyrrole-2,5-dione (1.8 mg, 0.0023 mmol) in EtOH (1 ml) was added copper sulfate pentahydrate (0.58 mg, 0.0023 mmol), VC (0.4 mg, 0.0023 mmol), THF (0.3 ml) and compound E2-6 (3 mg, 0.0023 mmol) and stirred at room temperature for 2 hours. The reaction was monitored to completion by LCMS. The reaction was purified by Prep-HPLC to give compound L42-E2 (1 mg, 20.5%) as a white solid. LC-MS (ESI): m / z found [1 / 2M+H] = 1059.6. +

[0735] Example 46: Synthesis of compound L2-E13

[0736] First Step: Synthesis of compound 2

[0737] To a solution of compound 1 (300 mg, 1.570 mmol) in DMF (5 ml) was added compound 1-1 (578 mg, 1.570 mmol) and HC1 / EA (1 ml) and stirred at room temperature for 3 hours. The reaction was monitored to completion by LCMS. The reaction was purified by reverse phase column chromatography to give compound 2 (200 mg, 25.5%) as a yellow solid. LC-MS (ESI): m / z found [M+H] = 1324.5. + ​​= 499.36.

[0738] Second Step: Synthesis of compound 3

[0739] Compound 2 (200 mg, 0.400 mmol) was dissolved in dichloromethane (5 ml), diethylamine (1 ml) was added, and the mixture was stirred at room temperature for 1.5 hours. The reaction was monitored by LCMS to be completed. The reaction was purified by reverse phase column chromatography to give compound 3 (120 mg, 99.8%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 277.16.

[0740] Third Step: Synthesis of compound 4

[0741] Compound 3 (120 mg, 0.434 mmol) was dissolved in AcOH (1 ml), Int. 1 (90 mg, 0.145 mmol) was added, and the mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS to be completed. The reaction was purified by HPLC to give compound 4 (50 mg, 13%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 880.30.

[0742] Fourth Step: Synthesis of compound L2-E13

[0743] Compound 4-2 (14.7 mg, 0.011 mmol) was dissolved in DMF (0.5 ml), HATU (6.47 mg, 0.017 mmol) was added, the system was cooled by ice bath and stirred for 20 min, compound 4 (10 mg, 0.011 mmol) and DIEA (2.20 mg, 0.017 mmol) were added, and the mixture was stirred at room temperature for 1.5 hours. The reaction was monitored by LCMS to be completed. The reaction was purified by HPLC to give compound L2-E13 (4 mg, 16%) as a yellow solid. LC-MS (ESI): m / z found [1 / 2M+H] + = 1081.32.

[0744] Example 47: Synthesis of compound L2-E31

[0745] First Step: Synthesis of compound 2

[0746] Compound 1 (500 mg, 2.403 mmol) was dissolved in DMF (10 ml), compound 1-1 (884 mg, 2.403 mmol) and HCl / EA (2 ml) were added, and stirred at room temperature for 3 hours. The reaction was monitored by LCMS to be completed. Compound 2 (400 mg, 32.1%) was obtained as a yellow solid after the reaction solution was purified by reverse phase column chromatography. LC-MS (ESI): m / z found [M+H]= 517.36. +

[0747] Second Step: Synthesis of compound 3

[0748] Compound 2 (400 mg, 0.773 mmol) was dissolved in dichloromethane (5 ml), diethylamine (1 ml) was added, and stirred at room temperature for 1.5 hours. The reaction was monitored by LCMS to be completed. Compound 3 (200 mg, 87.8%) was obtained as a white solid after the reaction solution was purified by reverse phase column chromatography. LC-MS (ESI): m / z found [M+H]= 295.16. +

[0749] Third Step: Synthesis of compound 4

[0750] Compound 3 (200 mg, 0.678 mmol) was dissolved in AcOH (1 ml), Int. 1 (150 mg, 2.034 mmol) was added, and stirred at room temperature for 16 hours after nitrogen replacement for three times. The reaction was monitored by LCMS to be completed. Compound 4 (100 mg, 16%) was obtained as a white solid after the reaction solution was purified by HPLC. LC-MS (ESI): m / z found [M+H]= 897.30. +

[0751] Fourth Step: Synthesis of compound L2-E31

[0752] Compound 4-2 (14.5 mg, 0.011 mmol) was dissolved in DMF (0.5 ml), HATU (6.47 mg, 0.017 mmol) was added, and the system was cooled by ice bath and stirred for 20 min, compound 4 (10 mg, 0.011 mmol) and DIEA (2.20 mg, 0.017 mmol) were added, and stirred at room temperature for 1.5 hours. The reaction was monitored by LCMS to be completed. Compound L2-E31 (5 mg, 17%) was obtained as a yellow solid after the reaction solution was purified by HPLC. LC-MS (ESI): m / z found [1 / 2M+H]= 1090.32. +

[0753] ​​​​Example 48: Synthesis of compound L44-E38

[0754] Step 1: Synthesis of compound L44-E38

[0755] L44 (18.7 mg, 0.0125 mmol) was dissolved in DMF (2 ml), HATU (5.7 mg, 0.015 mmol) was added in ice bath and reacted for 0.5 h at room temperature. E38 (10 mg, 0.0125 mmol), DIEA (5 ul) was added and stirred for 1 h at room temperature. The reaction was monitored by LCMS. Compound L44-E38 (9.3 mg, 32.2%) was obtained as a yellow solid after HPLC purification. LC-MS (ESI): m / z found [M / 2+H] = 1135.38. +

[0756] Example 49: Synthesis of compound L44-E39

[0757] Step 1: Synthesis of compound L44-E39

[0758] L44 (12.5 mg, 0.0083 mmol) was dissolved in DMF (2 ml), HATU (3.5 mg, 0.009 mmol) was added in ice bath and reacted for 0.5 h at room temperature. E39 (6 mg, 0.0076 mmol), DIEA (2.7 ul) was added and stirred for 1 h at room temperature. The reaction was monitored by LCMS. Compound L44-E39 (9.3 mg, 32.2%) was obtained as a yellow solid after HPLC purification. LC-MS (ESI): m / z found [M / 2+H] = 1130.38. +

[0759] Example 50: Synthesis of compound L44-E29

[0760] Step 1: Synthesis of compound L44-E29

[0761] ​​Compound E29 (2.6 mg, 0.0033 mmol) was dissolved in DMF (2 ml), L44 (5 mg, 0.0033 mmol), HATU (1.53 mg, 0.004 mmol), DIEA (0.87 mg, 0.0066 mmol) were added and stirred at room temperature for 1 hour. The reaction was monitored by LCMS. To the reaction was added EA (10 ml), washed with saturated sodium bicarbonate (20 ml), the organic phase was dried, filtered, concentrated and purified by HPLC to give compound L44-E29 (0.4 mg, 5.3%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=2255.4. +

[0762] Example 51: Synthesis of compound L2-E23

[0763] First step: Synthesis of compound 2

[0764] Compound 1 (430 mg, 1.963 mmol) was dissolved in DMF (12 ml), cooled in ice bath, compound 1-1 (722.5 mg, 1.963 mmol) and TFA (1.2 ml) were added, replaced with nitrogen for three times, stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction was purified by reverse phase column chromatography to give compound 2 (130 mg, 12.6%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=526.64. +

[0765] Second step: Synthesis of compound 3

[0766] Compound 2 (100 mg, 0.19 mmol) was dissolved in dichloromethane (5 ml), diethylamine (1 ml) was added, stirred at room temperature for 1.5 hours. The reaction was monitored by LCMS. The reaction was purified by reverse phase column chromatography to give compound 3 (56 mg, 96.8%) as a white solid. LC-MS (ESI): m / z found [M+H]=304.39. +

[0767] Third step: Synthesis of compound 4

[0768] ​​​Compound 3 (50 mg, 0.16 mmol) was dissolved in AcOH (1 ml), added Int. 1 (25 mg, 0.04 mmol), replaced with nitrogen for three times, stirred at room temperature for 16 hours. The reaction was monitored to completion by LCMS. The reaction was purified by HPLC to give compound 4 (10 mg, 27.5%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 908.04.

[0769] Fourth step: synthesis of compound L2-E23

[0770] Compound 4-2 (8.6 mg, 0.0066 mmol) was dissolved in DMF (0.5 ml), added HATU (3.78 mg, 0.0099 mmol), the system was cooled by ice bath and stirred for 20 min, added compound 4 (6 mg, 0.0066 mmol) and DIEA (1.28 mg, 0.0099 mmol), stirred at room temperature for 1.5 hours. The reaction was monitored to completion by LCMS. The reaction was purified by HPLC to give compound L2-E23 (2.2 mg, 15.2%) as a yellow solid. LC-MS (ESI): m / z found [1 / 2M+H] + = 1096.20.

[0771] Example 52: synthesis of compound L2-E41

[0772] First step: synthesis of compound 2

[0773] Compound 1 (3 g, 8.15 mmol) was dissolved in DCM (15 ml), cooled by ice bath, added compound 1-1 (3.99 g, 57.05 mmol) and TFA (3 ml), replaced with nitrogen for three times, stirred at room temperature for 2 hours. The reaction was monitored to completion by LCMS. The reaction was concentrated, then MeOH was added to the residual liquid, a white solid was precipitated, and compound 2 (2.8 g, 90.8%) was obtained as a white solid after filtration. LC-MS (ESI): m / z found [M+H] + = 378.43.

[0774] Second step: synthesis of compound 3

[0775] Compound 2 (3.3 g, 8.73 mmol) was dissolved in 1,4-dioxane (50 ml), TEA (8 ml), E23-4 (2.7 g, 8.73 mmol), cuprous iodide (500 mg, 2.62 mmol) and dichlorobis(triphenylphosphine)palladium (613 mg, 0.873 mmol) were added, and the reaction was stirred at room temperature for 3 hours under nitrogen. The reaction was monitored by LCMS. The reaction was poured into water (200 ml), and extracted with EA (100 ml*2). The organic phase was combined and dried, and concentrated. Purification was performed by normal phase column chromatography to obtain compound 3 (3.4 g, 68.7%) as a brown oil. LC-MS (ESI): m / z found [M+H] = 566.61. +

[0776] Step 3: Synthesis of compound 4

[0777] Compound 3 (1.7 g, 3.0 mmol) was dissolved in dichloromethane (51 ml), diethylamine (13 ml) was added, and the reaction was stirred at room temperature for 1.5 hours. The reaction was monitored by LCMS. Purification was performed by reverse phase column chromatography to obtain compound 4 (756 mg, 73.2%) as a white solid. LC-MS (ESI): m / z found [M+H] = 344.37. +

[0778] Step 4: Synthesis of compound 5

[0779] Compound 4 (300 mg, 0.871 mmol) was dissolved in MeOH (12 ml), Pd-C (900 mg, 300% wt) was added, and the reaction was stirred at room temperature for 16 hours under hydrogen. The reaction was monitored by LCMS. The reaction was filtered and concentrated to obtain compound 5 (140 mg, 50.5%) as a white solid. LC-MS (ESI): m / z found [M+H] = 318.42. +

[0780] Step 5: Synthesis of compound 6

[0781] Compound 5 (62 mg, 0.195 mmol) was dissolved in AcOH (1.5 ml), Int. 1 (40 mg, 0.064 mmol) was added, and the reaction was stirred at room temperature for 16 hours under nitrogen. The reaction was monitored by LCMS. Purification was performed by HPLC to obtain compound 6 (37 mg, 62.7%) as a white solid. LC-MS (ESI): m / z found [M+H] = 922.07. + ​​​​

[0782] Step 6: Synthesis of compound L2-E41

[0783] Compound 4-2 (21.2 mg, 0.0163 mmol) was dissolved in DMF (2 ml), HATU (9.3 mg, 0.0244 mmol) was added, the system was cooled by ice bath and stirred for 20 min, compound 6 (15 mg, 0.0163 mmol) and DIEA (3.15 mg, 0.0244 mmol) were added, and stirred at room temperature for 1.5 hours. The reaction was monitored by LCMS. The reaction was purified by HPLC to obtain compound L2-E41 (8.67 mg, 24.1%) as a yellow solid. LC-MS (ESI): m / z found [1 / 2M+H] + = 1103.22.

[0784] Example 53: Synthesis of compound L3-E13

[0785] Step 1: Synthesis of compound L3-E13-2

[0786] Compound L3-E13-1 (280 mg, 0.68 mmol) was dissolved in DMF (5 ml), HOSU (117 mg, 1.01 mmol) and EDCI (195 mg, 1.01 mmol) were added, and stirred at room temperature for 3 hours. (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-aminobutanoic acid (500 mg, 1.47 mmol) was added, and the reaction was monitored by LCMS. After the reaction was completed, the reaction was purified by reverse phase column chromatography to obtain compound L3-E13-2 (181 mg, 45.5%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + = 911.36.

[0787] Step 2: Synthesis of compound L3-E13-3

[0788] Compound L3-E13-2 (181 mg, 0.20 mmol) was dissolved in DMF (5 ml), HOSU (34 mg, 0.20 mmol) and DCC (61 mg, 0.20 mmol) were added, stirred at room temperature for 3 hours. LCMS was used to monitor the completion of the reaction. Compound L3-E13-3 (185 mg, 75.5%) was obtained as a yellow solid after the reaction solution was purified by reverse phase column chromatography. LC-MS (ESI): m / z found [M+H]=1171.58. +

[0789] Third step: synthesis of compound L3-E13-4

[0790] Compound L3-E13-3 (185 mg, 0.024 mmol) was dissolved in DCM (3 ml), diethylamine (1 ml) was added, replaced with nitrogen for three times, stirred at room temperature for 16 hours. LCMS was used to monitor the completion of the reaction. Compound L3-E13-4 (50 mg, 23%) was obtained as a white solid after the reaction solution was purified by HPLC. LC-MS (ESI): m / z found [M+H]=950.30. +

[0791] Fourth step: synthesis of compound L3-E13-5

[0792] Compound L3-E13-4 (50 mg, 0.011 mmol) was dissolved in DMF (0.5 ml), HATU (2.8 mg, 0.017 mmol) was added, the system was cooled by ice bath and stirred for 20 min, compound 3-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)propanoic acid (4.8 mg, 0.011 mmol) and DIEA (3.8 mg, 0.017 mmol) were added, stirred at room temperature for 1.5 hours. LCMS was used to monitor the completion of the reaction. Compound L3-E13-5 (30 mg, 16%) was obtained as a yellow solid after the reaction solution was purified by HPLC. LC-MS (ESI): m / z found [M+H]=1179.33. +

[0793] Fifth step: synthesis of compound L3-E13-6

[0794] ​​​Compound L3-E13-5 (30 mg, 0.011 mmol) was dissolved in DMF (0.5 ml), HATU (1.8 mg, 0.017 mmol) was added, the system was cooled by ice bath and stirred for 20 min, compound E13 (4 mg, 0.011 mmol) and DIEA (2.8 mg, 0.017 mmol) were added, and stirred at room temperature for 1.5 hours. The reaction was monitored by LCMS to be completed. The reaction was purified by HPLC to give compound L3-E13-6 (20 mg, 46%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=2083.38. +

[0795] Step 6: Synthesis of compound L3-E13

[0796] Compound L3-E13-5 (20 mg, 0.011 mmol) was dissolved in DMSO (0.5 ml), VC (2 mg, 0.017 mmol) was added, anhydrous copper sulfate (1.8 mg, 0.017 mmol) was replaced by nitrogen three times, and stirred at room temperature for 1.5 hours. The reaction was monitored by LCMS to be completed. The reaction was purified by HPLC to give compound L3-E13 (5 mg, 26%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=2077.38. +

[0797] Example 54: Synthesis of compound L3-E31

[0798] Step 1: Synthesis of compound L3-E31-2

[0799] Compound L3-E31-1 (20 mg, 0.011 mmol) was dissolved in DMF (0.5 ml), HATU (1.8 mg, 0.017 mmol) was added, the system was cooled by ice bath and stirred for 20 min, compound E31 (4 mg, 0.011 mmol) and DIEA (2.8 mg, 0.017 mmol) were added, and stirred at room temperature for 1.5 hours. The reaction was monitored by LCMS to be completed. The reaction was purified by HPLC to give compound L3-E31-2 (10 mg, 44%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=2099.97. +

[0800] Step 6: Synthesis of compound L3-E31

[0801] ​​​Compound L3-E31-2 (10 mg, 0.011 mmol) was dissolved in DMSO (0.5 ml), VC (2 mg, 0.017 mmol) was added, anhydrous copper sulfate (1.8 mg, 0.017 mmol) was added, and the mixture was stirred at room temperature for 1.5 hours. The reaction was monitored by LCMS. The reaction mixture was purified by HPLC to give compound L3-E31 (2 mg, 11%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=2853.1. + = 2853.1.

[0802] Example 55: Synthesis of compound L3-E23

[0803] Step 1: Synthesis of compound L3-E23

[0804] Compound L3 (7 mg, 0.00355 mmol) was dissolved in DMF (0.5 ml), HATU (2.5 mg, 0.0066 mmol) was added, the mixture was cooled in an ice bath and stirred for 20 min, compound E23-int.1 (4 mg, 0.0066 mmol) and DIEA (0.85 mg, 0.0066 mmol) were added, and the mixture was stirred at room temperature for 1.5 hours. The reaction was monitored by LCMS. The reaction mixture was purified by HPLC to give compound L2-E23 (1 mg, 9.8%) as a yellow solid. LC-MS (ESI): m / z found [1 / 2M+H]=1431.60. + = 1431.60.

[0805] Example 56: Synthesis of compound L4-E29

[0806] Step 1: Synthesis of compound L4-E29-1

[0807] Compound E29 (100 mg, 0.127 mmol) was dissolved in DMF (2 ml), tert-butyloxycarbonyl-L-alanine (24.2 mg, 0.127 mmol), HATU (72.9 mg, 0.191 mmol), and DIEA (24.7 mg, 0.191 mmol) were added, and the mixture was stirred at room temperature for 1 hour. The reaction was monitored by LCMS. To the reaction mixture was added EA (10 ml), and the mixture was washed with saturated sodium bicarbonate (20 ml). The organic phase was dried, filtered, and concentrated, and the residue was purified by HPLC to give compound L4-E29-1 (30 mg, 24.6%) as a white solid. LC-MS (ESI): m / z found [M+H]=954.0. + = 954.0.

[0808] Second Step: Synthesis of compound L4-E29-2

[0809] Compound L4-E29-1 (30 mg, 0.031 mmol) was dissolved in DCM (2 ml), HCl / dioxane (0.5 ml) was added, stirred at room temperature for 1 hour. LCMS monitored the completion of the reaction. The reaction was concentrated and dried to get compound L4-E29-2 (15 mg, 55.8%) as white solid. LC-MS (ESI): m / z found [M+H]+ = 854.0. +

[0810] Third Step: Synthesis of compound L4-E29-3

[0811] Compound L4-E29-2 (15 mg, 0.017 mmol) was dissolved in DMF (2 ml), (1-azido-3,6,9,12-tetraoxypentadecan-15-oyl)-L-valine (6.9 mg, 0.017 mmol), HATU (9.9 mg, 0.026 mmol), DIEA (4.5 mg, 0.034 mmol) were added, stirred at room temperature for 1 hour. LCMS monitored the completion of the reaction. To the reaction was added EA (10 ml), saturated sodium bicarbonate (20 ml) was washed, the organic phase was dried, filtered, concentrated and purified by HPLC to get compound L4-E29-3 (10 mg, 46.4%) as white solid. LC-MS (ESI): m / z found [M+H]+ = 1226.3.

[0812] Fourth Step: Synthesis of compound L4-E29

[0813] Compound L4-E29-3 (10 mg, 0.008 mmol) was dissolved in DMSO (2 ml), linker (5.2 mg, 0.006 mmol), VC (1.4 mg, 0.008 mmol) and copper sulfate pentahydrate (2 mg, 0.008 mmol) were added, stirred at room temperature for 1 hour. LCMS monitored the completion of the reaction. The reaction was purified by HPLC to get compound L4-E29 (1.2 mg, 7.3%) as yellow solid. LC-MS (ESI): m / z found [M+H]+ = 2020.2. +

[0814] Example 57: Synthesis of compound L4-E44

[0815] First Step: Synthesis of compound L4-E44-2 ​​

[0816] Compound L4-E44-1 (20 mg, 0.016 mmol) was dissolved in ACN (1.5 ml) and H2O (1 ml), silver nitrate (52.7 mg, 0.310 mmol) was added, stirred at room temperature for 12 hours. LCMS monitoring reaction completion. To the reaction solution was added saturated brine (2.5 ml) and saturated sodium bicarbonate (2.5 ml), stirred for 15 min, the mixture was extracted with EA (5 ml*2) and water (10 ml), the organic phase was combined and dried over anhydrous sodium sulfate, concentrated and purified by HPLC to give compound L4-E44-2 (7 mg, 35.3%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 1217.3.

[0817] Second Step: Synthesis of compound L4-E44

[0818] Compound E44-L4-2 (7 mg, 0.005 mmol) was dissolved in DMSO (2 ml), linker (3.7 mg, 0.004 mmol), VC (1.0 mg, 0.005 mmol) and copper sulfate pentahydrate (1.4 mg, 0.005 mmol) were added, stirred at room temperature for 1 hour. LCMS monitoring reaction completion. The reaction was purified by HPLC to give compound L4-E44 (0.5 mg, 4.3%) as a yellow solid. LC-MS (ESI): m / z found [M+H] + = 1967.1.

[0819] Example 58: Synthesis of compound L5-E23

[0820] First Step: Synthesis of compound L5-2

[0821] Compound L5-1 (1.5 g, 11.36 mmol) was dissolved in THF (40 ml), replaced with nitrogen three times, cooled in an ice bath, LAH (13.5 ml) was added, and stirred at 70°C for 4 hours. LCMS monitoring reaction completion. The reaction was purified by reversed-phase column chromatography, concentrated to give compound L5-2 (700 mg, 45.2%) as a white solid. LC-MS (ESI): m / z found [M+H] + = 136.20.

[0822] Second Step: Synthesis of compound L5-3

[0823] Compound L5-2 (533 mg, 3.92 mmol) was dissolved in ACN (15 ml), triethylamine (435.5 mg, 4.312) and di-tert-butyl dicarbonate (897 mg, 4.11 mmol) were added, stirred at room temperature for 2 hours. LCMS monitored the reaction was completed. The reaction was purified by reverse phase column chromatography, concentrated to give compound L5-3 (230 mg, 24.8%) as a white solid. LC-MS (ESI): m / z found [M+H]= 236.32. + = 236.32.

[0824] Step 3: Synthesis of compound L5-3-E23

[0825] Compound E23 (50 mg, 0.06 mmol) was dissolved in THF (1 ml), DIEA (9.5 mg, 0.073 mmol) and triphosgene (21.7 mg, 0.073 mmol) were added, stirred at room temperature for 1.5 hours, DIEA (14.6 mg, 0.113 mmol) and compound L5-3 (13.34 mg, 0.06 mmol) were added, stirred at room temperature for 1 hour. LCMS monitored the reaction was completed. The reaction was purified by normal phase column chromatography (PE / EA), concentrated to give compound L5-3-E23 (30 mg, 46.1%) as a light yellow solid. LC-MS (ESI): m / z found [M+H]= 1084.26. + = 1084.26.

[0826] Step 4: Synthesis of compound L5-4-E23

[0827] Compound L5-3-E23 (30 mg, 0.0276 mmol) was dissolved in DCM (0.5 ml), TFA (0.1 ml) was added, stirred at room temperature for 2 hours. LCMS monitored the reaction was completed. The reaction was purified by reverse phase column chromatography, lyophilized to give compound L5-4-E23 (5 mg, 18.4%) as a white solid. LC-MS (ESI): m / z found [M+H]= 984.14. + = 984.14.

[0828] Step 5: Synthesis of compound L5-E23

[0829] Compound L42 (7.2 mg, 0.005 mmol) was dissolved in DMF (0.5 ml), the system was cooled in ice bath, L5-4-E23 (5 mg, 0.05 mmol) and DIEA (1 mg, 0.0075 mmol) were added, and it was stirred at room temperature for 1.5 hours. The reaction was monitored by LCMS to be completed. The reaction was purified by HPLC to obtain compound L5-E23 (1.6 mg, 14.1%) as a yellow solid. LC-MS (ESI): m / z found [1 / 2M+H] + = 1133.25.

[0830] Example 59: Synthesis of compound L6-E48

[0831] First step: Synthesis of compound L6-2

[0832] Compound L6-1 (600 mg, 0.659 mmol) was dissolved in DMF (6 ml), cooled in ice bath, HOSU (90.9 mg, 0.79 mmol) and EDCI (151.4 mg, 0.79 mmol) were added, stirred at room temperature for 3 hours, DIEA (127.5 mg, 0.988 mmol) and L6-1A (278.6 mg, 0.659 mmol) were added, stirred at room temperature for 1 hour. The reaction was monitored by LCMS to be completed. The reaction was purified by reverse phase column chromatography, and after concentration, compound L6-2 (700 mg, 80.7%) was obtained as a white solid. LC-MS (ESI): m / z found [M+H] + = 1316.46.

[0833] Second step: Synthesis of compound L6-3

[0834] Compound L6-2 (700 mg, 0.532 mmol) was dissolved in DCM (8.5 ml), diethylamine (4.2 ml) was added, and it was stirred at room temperature for 2 hours. The reaction was monitored by LCMS to be completed. The reaction was purified by reverse phase column chromatography, and after concentration, compound L6-3 (410 mg, 70.4%) was obtained as a white solid. LC-MS (ESI): m / z found [M+H] + = 1094.22.

[0835] Third step: Synthesis of compound L6-4

[0836] Compound L6-3 (80 mg, 0.0732 mmol) was dissolved in DMF (1 ml), DIEA (18.8 mg, 0.146 mmol) and azidoacetic acid NHS ester (29 mg, 0.146 mmol) were added, stirred at room temperature for 1.5 hours. LCMS monitored the completion of the reaction. The reaction was purified by reverse phase column chromatography, after freeze-drying to obtain compound L6-4 (40 mg, 46.4%) as a white solid. LC-MS (ESI): m / z found [M+H]=1177.27. +

[0837] Fourth step: synthesis of compound L6

[0838] Compound L6-4 (20 mg, 0.017 mmol) was dissolved in DMSO (12 ml), copper sulfate pentahydrate (4.25 mg, 0.017 mmol), ascorbic acid (3 mg, 0.017 mmol) and L6-4A (13.5 mg, 0.017 mmol) were added, stirred at room temperature for 2 hours. LCMS monitored the completion of the reaction. The reaction was purified by reverse phase column chromatography, after freeze-drying to obtain compound L6 (10 mg, 29.8%) as a white solid. LC-MS (ESI): m / z found [M+H]=1971.12. +

[0839] Fifth step: synthesis of compound L6-E48

[0840] Compound L6 (10 mg, 0.005 mmol) was dissolved in ACN (0.5 ml), the system was cooled by ice bath and stirred for 20 min, N-methylimidazole (4.1 mg, 0.05 mmol) and E48 (4 mg, 0.005 mmol) were added, TCFH (3.5 mg, 0.0125 mmol) was added, stirred at room temperature for 1.5 hours. LCMS monitored the completion of the reaction. The reaction was purified by HPLC to obtain compound L6-E48 (1.5 mg, 10.9%) as a yellow solid. LC-MS (ESI): m / z found [1 / 2M+H]=1366.51. +

[0841] Example 60: synthesis of compound L6-E13

[0842] First step: synthesis of compound L6-E13

[0843] ​​​Compound L6 (20 mg, 0.011 mmol) was dissolved in DMF (0.5 ml), HATU (1.2 mg, 0.017 mmol) was added, the system was cooled by ice bath and stirred for 20 min, compound E13 (7 mg, 0.011 mmol) and DIEA (1.8 mg, 0.017 mmol) were added, and stirred at room temperature for 1.5 hours. The reaction was monitored by LCMS to be completed. The reaction was purified by HPLC to obtain compound L6-E13 (7 mg, 19%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=2206.4. +

[0844] Example 61: Synthesis of compound L7-E13

[0845] First step: Synthesis of compound L7-E13

[0846] Compound L7 (27.5 mg, 0.017 mmol) was dissolved in DMF (2 ml), E13 (15 mg, 0.017 mmol), HATU (7.8 mg, 0.020 mmol), DIEA (4.4 mg, 0.020 mmol) were added, and stirred at room temperature for 1 hour. The reaction was monitored by LCMS to be completed. After concentration, HPLC purification was performed to obtain compound L7-E13 (10 mg, 26.5%) as a yellow solid. LC-MS (ESI): m / z found [M+H]=2209.4. +

[0847] Example 62: Preparation of antibodies

[0848] The following antibodies were prepared using conventional methods, the heavy chain and light chain plasmids of the antibodies were transfected into 293f cells, and the sample was collected after 4 days, the supernatant was collected by high-speed centrifugation, and purified using a Protein A column.

[0849] Nectin-4 antibody-1 (prepared according to WO2025162492A1)

[0850] Heavy chain amino acid sequence: (SEQ ID NO: 1)

[0851] Light chain amino acid sequence: (SEQ ID NO: 2)

[0852] Heavy chain CDR amino acid sequence

[0853] HCDR1 (SEQ ID NO: 11): TYDVH

[0854] ​​HCDR2 (SEQ ID NO: 12): VIWSGGSTDFNAAFRS

[0855] HCDR3 (SEQ ID NO: 13): DRYYAMEY

[0856] Heavy chain CDR amino acid sequences

[0857] HCDR1 (SEQ ID NO: 19): SYNMN

[0858] HCDR2 (SEQ ID NO: 20): YISSSSSTIYYADSVKG

[0859] HCDR3 (SEQ ID NO: 21): AYYYGMDV

[0860] Heavy chain variable region amino acid sequence: (SEQ ID NO: 17)

[0861] Light chain variable region amino acid sequence: (SEQ ID NO: 18)

[0862] Nectin4 Antibody-2 (prepared as per WO2012047724)

[0863] Heavy chain amino acid sequence: (SEQ ID NO: 3)

[0864] Light chain amino acid sequence: (SEQ ID NO: 4)

[0865] Heavy chain CDR amino acid sequences

[0866] HCDR1 (SEQ ID NO: 19): SYNMN

[0867] HCDR2 (SEQ ID NO: 20): YISSSSSTIYYADSVKG

[0868] HCDR3 (SEQ ID NO: 21): AYYYGMDV

[0869] Light chain CDR amino acid sequences

[0870] LCDR1 (SEQ ID NO: 22): RASQGISGWLA

[0871] LCDR2 (SEQ ID NO: 23): AASTLQS

[0872] LCDR3 (SEQ ID NO:24): QQANSFPPT

[0873] Heavy chain variable region amino acid sequence: (SEQ ID NO: 25)

[0874] Heavy chain variable region amino acid sequence: (SEQ ID NO: 25)

[0875] Nectin4 Antibody-3 (prepared according to CN117942409A)

[0876] Heavy chain amino acid sequence: (SEQ ID NO: 5)

[0877] Heavy chain amino acid sequence: (SEQ ID NO: 5)

[0878] Heavy chain CDR amino acid sequences

[0879] HCDR1 (SEQ ID NO:27): DYGVS

[0880] HCDR2 (SEQ ID NO:28): VIWGGGKIYYNSVLK

[0881] HCDR3 (SEQ ID NO:29): QGGLLFYAMDY

[0882] Light chain CDR amino acid sequences

[0883] LCDR1 (SEQ ID NO:30): KSSQSLLNTYSQKNYLA

[0884] LCDR2 (SEQ ID NO:31): FASTRES

[0885] LCDR3 (SEQ ID NO:32): QQHYNTPFT Heavy chain variable region amino acid sequence: (SEQ ID NO: 33)

[0886] Light chain variable region amino acid sequence: (SEQ ID NO: 34)

[0887] B7H3 Antibody-1

[0888] Heavy chain amino acid sequence: (SEQ ID NO: 7)

[0889] Heavy chain amino acid sequence: (SEQ ID NO: 9)

[0890] Heavy chain CDR amino acid sequences

[0891] HCDR1 (SEQ ID NO: 35): SYAMS

[0892] HCDR2 (SEQ ID NO: 36): SISGNGKYIYYLDSVKG HCDR3 (SEQ ID NO: 37): HYDGYLDV

[0893] Light chain CDR amino acid sequences

[0894] LCDR1 (SEQ ID NO: 38): RASDIIYNYLA

[0895] LCDR2 (SEQ ID NO: 39): NANTLGE

[0896] LCDR3 (SEQ ID NO: 40): QHHYGTPTWT

[0897] Heavy chain variable region amino acid sequence: (SEQ ID NO: 41)

[0898] Light chain variable region amino acid sequence: (SEQ ID NO: 42)

[0899] B7H3 antibody-2 (prepared according to CN107922477B)

[0900] Heavy chain amino acid sequence: (SEQ ID NO: 9)

[0901] Light chain amino acid sequence: (SEQ ID NO: 10)

[0902] Heavy chain CDR amino acid sequences

[0903] HCDR1 (SEQ ID NO: 43): NYVMH

[0904] HCDR2 (SEQ ID NO: 44): YINPYNDDVKYNEKFKG

[0905] HCDR3 (SEQ ID NO: 45): WGYYGSPLYYFDY

[0906] Light chain CDR amino acid sequences

[0907] LCDR1 (SEQ ID NO: 46): RASSRLIYMH

[0908] LCDR2 (SEQ ID NO: 47): ATSNLAS

[0909] LCDR3 (SEQ ID NO: 48): QQWNSNPPT

[0910] Heavy chain variable region amino acid sequence: (SEQ ID NO: 49)

[0911] Light chain variable region amino acid sequence: (SEQ ID NO: 50)

[0912] Example 63: Preparation of ADC

[0913] Under room temperature conditions, 1 mM EDTA-PBS buffer solution, appropriate amount of antibody, 5 mM tris (2-hydroxyethyl) phosphine hydrochloride (TCEP) stock solution, the final concentration of the reaction system is 4 mg / ml, the molar ratio of TCEP to antibody is 5, after fully mixing, it is placed in a 37°C, 220 rpm shaker for 2 hours of reduction reaction. After the reduction is completed, DMSO is added to the coupling reaction DMSO final volume of 15%, then LP is added, the molar ratio of LP to antibody is 5, after fully mixing, it is placed in a constant temperature mixing instrument, and the reaction is carried out at room temperature for 3-4 hours. After the coupling is completed, the ADC is desalted and purified by Zeba TM Desalting centrifugal column, 7K MWCO, to obtain ADC stock solution, which is stored at -20°C after aliquoting. The prepared ADC structure is shown in the following table.

[0914] Table 1: ADC structure

[0915] Note: The ADC maleimide ring has two forms of ring opening and ring closing. In the preparation process, part or all of the maleimide linker in the ADC molecule will hydrolyze, and (closed ring) to (open ring).

[0916] Test Example 1: Determination of ADC-DAR value

[0917] HIC determination method:

[0918] Sample processing: The ADC prepared above is diluted to a concentration of 2 mg / ml using PBS, and the sample volume is 10 uL.

[0919] SEC assay method

[0920] Sample treatment: The ADC prepared above was diluted to a concentration of 2 mg / ml with PBS, and 10 uL of the sample was injected.

[0921] HIC elution gradient condition

[0922] TOF assay method

[0923] Sample treatment: The ADC prepared above was diluted to 1 mg / mL with PBS, filtered with a 0.22 um filter, and then 10 uL of 1 M DTT was added to each 120 samples, and after mixing, the sample was injected.

[0924] Liquid phase model: LC-40AD Shimadzu

[0925] Mobile phase A: 1000 mL of purified water was measured with a measuring cylinder, 1 mL of formic acid was added, and it was ultrasonically prepared for use, with a validity period of 7 days.

[0926] Mobile phase B: 1000 mL of acetonitrile was measured with a measuring cylinder, 1 mL of formic acid was added, and it was ultrasonically prepared for use, with a validity period of 7 days.

[0927] Needle washing liquid: 500 mL of purified water was measured with a measuring cylinder, 500 mL of methanol was measured with a measuring cylinder, and after mixing, it was ultrasonically prepared for use, with a validity period of 7 days.

[0928] The DAR value information of the ADC prepared above is described in the following table: - indicates not tested

[0929] Test example 2: Payload activity test

[0930] a. Cell suspensions were prepared with fresh cell culture medium containing 10% FBS, NCI-H358, NCI-H345, PANC-1, LNCaP, DU145, DLD-1, GP2d, MDA-MB-453 were added to 96-well cell culture plates (Fisher scientific: 2308025) at 100 uL of 2000 cells per well, A375 was added to 96-well cell culture plates (Fisher scientific: 2308025) at 100 uL of 1000 cells per well, VCAP was added to 96-well cell culture plates (Fisher scientific: 2308025) at 100 uL of 40000 cells per well, T84 was added to 96-well cell culture plates (Fisher scientific: 2308025) at 100 uL of 4000 cells per well, MIA PaCa-2 and NCI-H526 were added to 96-well cell culture plates (Fisher scientific: 2308025) at 50 uL of 4000 cells per well, and PBS was added to the outermost wells of the cell plate, and incubated at 37°C under 5% CO2 for 24 hours.

[0931] b. Prepare the medium solution containing Payload with the first hole concentration of 1200 nM, and dilute it with fresh medium by 5 times step by step, totally 10 concentration points (the 10th point is zero concentration point), and incubate for 5 days at 37℃ with 5% CO2.

[0932] c. Discard the old medium on the last day, and add 100 μL medium containing CCK8 detection solution (blank medium: CCK8 ratio is 9:1, CCK8 detection solution: C0039) to each hole, and add 10 μL CCK8 detection solution to each hole for MIA PaCa-2 and NCI-H526, and continue to incubate for 4 hours at 37℃ with 5% CO2.

[0933] d. Measure the absorbance at 450 nm by using the enzyme marker.

[0934] e. Process and analyze the results by using Graphpad Prism.

[0935] f. Different cell culture medium types

[0936] The experimental results are shown in Table 2 and Table 3.

[0937] Table 2 Anti-proliferation ability (IC 50 , unit: nM) of the compound of the present disclosure on various tumor cell lines Note: - means not tested.

[0938] Table 3 Anti-proliferation ability (IC 50 , unit: nM) of the compound of the present disclosure on various tumor cell lines

[0939] The results show that the Payload compound provided by the present disclosure has strong killing effect on various tumor cells.

[0940] Test Example 3: ADC activity test

[0941] Instruments and reagents

[0942] Experimental steps:

[0943] (1) Prepare a cell suspension using 1640 medium containing 10% FBS, and inoculate NCI-H526 / LNCap / DU145 / NCI-H345 cells into a 96-well plate at a density of 5×10 3 / 2×10 3 / 2×10 3 cells / 100 μL, add 200 μL PBS to the outermost circle, and incubate in a 37℃, 5% CO2 incubator for 24 h.

[0944] (2) Prepare the ADC-containing medium solution, the initial concentration of LNCAP cells in the 96-well plate is 1500 nM (or 3000 nM), and 10-fold gradient dilution is performed. Discard the old culture medium in the 96-well cell plate, and add 100 μL of the diluted drug to each well; the initial concentration of NCI-H526 / DU145 / NCI-H345 cells in the 96-well plate is 3000 nM, and 50 μL of the diluted drug is added to each well; 8 gradients of dilution are prepared, and the 8th well is fresh medium without drug. Incubate at 37°C in a 5% CO2 incubator for 5 days.

[0945] (3) On the 5th day, remove the 96-well plate, discard the culture solution of the LNCAP cell plate, and add 100 μL of detection solution (CCK8: medium = 1:9) to each well. Add 10 μL of CCK8 stock solution directly to each well of the NCI-H526 / DU145 / NCI-H345 cell plate; place in a 37°C, 5% CO2 incubator for 4 h.

[0946] (4) Read the values at 450 nm using an enzyme-labeled instrument.

[0947] (5) Analyze and organize the data using Graphpad Prism 8.0. The results are shown in Tables 4-8.

[0948] Table 4 Inhibitory effect of exemplary ADC on tumor cells (%)

[0949] Table 5

[0950] Table 6

[0951] Table 7

[0952] Table 8

[0953] The results show that the ADC of the present disclosure can produce a strong inhibitory effect on different tumor cells.

[0954] The present application has been described by the above examples, but it should be understood that the above examples are only for the purpose of example and illustration, and are not intended to limit the present application to the scope of the described examples. In addition, those skilled in the art can understand that the present application is not limited to the above examples, and more variations and modifications can be made according to the teachings of the present application, which all fall within the scope of the present application. The scope of protection of the present application is defined by the attached claims and their equivalent scope.

Claims

A compound as shown in Formula I-1, or an isomer, isotopic derivative, polymorph, prodrug, pharmaceutically acceptable salt, or solvate thereof, wherein, R1and R2are each independently selected from H, halogen, hydroxyl, C(=0)R0, C(=0)OR0, -OC(=0)OR0, -OC(=0)R0, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted C1-C6alkoxy, and substituted or unsubstituted C1-C6haloalkyl; said C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C1-C6alkoxy, and C1-C6haloalkyl are optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, and Y9are each independently selected from H, C1-C6alkyl, C1-C6alkoxy, C1-C6haloalkyl, OH, NO2, NH2, SH, CN, or halogen; X1is independently at each occurrence selected from the group consisting of halogen, alkyl, haloalkyl, OR x , CN, (=0) and H; wherein R x is selected from the group consisting of H, alkyl, haloalkyl, alkenyl, alkynyl, OH, OR0, SR0, SOR0and SO2R0; R3is selected from CR 3a R 3b ; R 3a is H, alkyl, alkoxy, OH, NO2, NH2, SH, CN or halogen; R 3b selected from R 3aa for R 3ab selected from OH, and N(R 00 )2; R 3b Not R4is selected from C(R 4a )2; R 4a each occurrence is independently selected from the group consisting of H, alkyl, alkoxy, OH, NO2, NH2, SH, CN, and halogen; or R3 is CR 3a , R4 is CR 4a , and R3 and R4 form, together with the atoms to which they are attached, a Q1is each independently for each occurrence selected from NR 00 , C(R 00 )2、 O and S; Q2is each independently for each occurrence selected from a single bond, NR 00 , C(R 00 )2、 R a1 and R a2 each independently is selected from H, OR0, SR0, SOR0, SO2R0, C(=0)R0, C(=0)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; R5and R6are each independently selected from H, halogen, C(=0)R0, C(=0)OR0, substituted or unsubstituted C1-C6alkyl, C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl; said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, and C2-C6alkynyl are optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; Q4is CR0or N, and Q3is CR0or N; Q6is C; Q5is NR0; R c1 , R c2 , R c3 , R c4 , R c6 , R c7 , and R c8 are each independently selected from R c0 , H, OH, -NO2, -NH2, -SH, -CN, halogen, -C(=O)H, -C(=O)CH3, -CO2H, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxyl, substituted or unsubstituted C2-C6alkenyl, and substituted or unsubstituted C2-C6alkynyl; said C1-C6alkyl, C1-C6alkoxyl, C2-C6alkenyl, and C2-C6alkynyl are optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; R c5 selected from R c0 , H, OH, -N02, -NH2, -SH, -CN, halogen, -C(=0)H, -C(=0)CH3, -C02H, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, and substituted or unsubstituted C2-C6alkynyl; said Ci-C6alkyl, C2-C6alkenyl, and C2-C6alkynyl are optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and Ci-C6alkyl; R c5 not R c9 For R c0 , and R c9 is not and R6is H, R c9 is R c0 For R ca -(R cb ) m0 -, R ca selected from R cb each occurrence is independently selected from -C(O)-, -N(R0)-, -O-, -C(S)-, -SO2-, -C(R0)2-; m0is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; m1is each occurrence independently selected from 0, 1, 2, 3, 4, 5, 6, and 7; m2is each occurrence independently selected from 1, 2, 3, 4, 5, and 6; R3 is CR 3a , R4 is CR 4a , and R3 and R4 form, with the atoms to which they are attached, formula 1-F: when R c6 is H, R c5 is not H, methoxy, hydroxy, methyl, -NH-methyl, F, -NHCOCH2OH; or when R c5 is amino, R c6 is not methoxy; X2, X3, X4, X5, and X6are each occurrence independently selected from H, OR0, SR0, SOR0, SO2R0, C(=0)R0, C(=0)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, C1-C6alkoxy, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl are optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; R0and R 00 each occurrence is independently selected from H, OH, N02, NH2, SH, CN, halogen, -C(=0)H, -C(=0)-Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; said Ci-C6alkyl, Ci-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl are optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and Ci-C6alkyl; Cy is selected from the group consisting of 3-10 membered cycloalkylene, 3-10 membered heterocyclylene containing at least one heteroatom independently selected from N, O, and S, 6-10 membered arylene, and 5-10 membered heteroarylene containing at least one heteroatom independently selected from N, O, and S, the 3-10 membered cycloalkylene, 3-10 membered heterocyclylene, 6-10 membered arylene, and 5-10 membered heteroarylene optionally substituted with a substituent selected from the group consisting of deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, hydroxyl, -NO2, -NH2, and -CN; and n each occurrence is independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. A compound of Formula I or a pharmaceutically acceptable salt thereof, wherein R1and R2are each independently selected from H, halogen, hydroxyl, C(=O)R0, C(=O)OR0, -OC(=O)OR0, -OC(=O)R0, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted C1-C6alkoxy, and substituted or unsubstituted C1-C6haloalkyl; preferably, R1and R2are each independently selected from H, halogen, hydroxyl, C(=O)R0, C(=O)OR0, -OC(=O)OR0, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted C1-C6alkoxy, and substituted or unsubstituted C1-C6haloalkyl, said C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C1-C6alkoxy, and C1-C6haloalkyl are optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; more preferably, R1is -OC(=O)CH3; and / or, R2is methyl; Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, and Y9are each independently selected from H, C1-C6alkyl, C1-C6alkoxy, C1-C6haloalkyl, OH, NO2, NH2, SH, CN, or halogen; preferably, Y1is OH, and / or, Y2is -OCH3, and / or Y3is CH3, and / or, Y4, Y5, Y6, Y7, Y8, and Y9are each independently H; X1is at each occurrence independently selected from the group consisting of halogen, alkyl, haloalkyl, OR x , CN, (=0) and H; wherein R x is selected from the group consisting of H, alkyl, haloalkyl, alkenyl, alkynyl, OH, OR0, SR0, SOR0and SO2R0; preferably X1is selected from the group consisting of hydroxyl and CN; R3is selected from CR 3a R 3b ; R 3a is H, alkyl, alkoxy, OH, NO2, NH2, SH, CN or halogen; R 3b selected from R 3aa for R 3ab selected from OH, and N(R 00 )2; R 3b not R4is selected from C(R 4a )2; R 4a each occurrence is independently selected from the group consisting of H, alkyl, alkoxy, OH, NO2, NH2, SH, CN, and halogen; or R3 is CR 3a , R4 is CR 4a , and R3 and R4 form, together with the atoms to which they are attached, a Q1and Q2are each independently selected from C(R 00 )2、 O and S; R a1 , and R a2 are each independently selected from H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; preferably, R a1 , and R a2 each is independently selected from the group consisting of H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, C1-C6alkoxy, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, C1-C6alkoxy, aryl, and heteroaryl are optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; when neither Q1nor Q2is R a1 and R a2 at least one of when at least one of Q1and Q2is R a1 and R a2 each independently is selected from the group consisting of H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, NH2, NHR0, N(R0)2, NHC(O)R0, CN, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; preferably, R a1 and R a2 each independently is selected from the group consisting of H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, C1-C6alkoxy, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, C1-C6alkoxy, aryl, and heteroaryl are optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C6alkyl. R5and R6are each independently selected from the group consisting of H, halogen, C(=0)R0, C(=0)OR0, substituted or unsubstituted C1-C6alkyl, C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl; preferably, R5and R6are each independently selected from the group consisting of H, halogen, C(=0)R0, C(=0)OR0, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, and substituted or unsubstituted C2-C6alkynyl, said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, and C2-C6alkynyl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; Q4is CR0or N, and Q3is CR0or N; preferably, Q4is CR0or N, and Q3is CR0or N; and at least one of Q3and Q4is N; more preferably, Q4is CH or N, and Q3is CH or N, and at least one of Q3and Q4is N; Q6is C; Q5is NR0; preferably, Q5is NH; R c1 , R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , and R c8 are each independently selected from R c0 , H, OH, -N02, -NH2, -SH, -CN, halogen, -C(=0)H, -C(=0)CH3, -C02H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C2-C6 alkenyl, and substituted or unsubstituted C2-C6 alkynyl; preferably, R c1 , R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , and R c8 are each independently selected from R c0 , H, OH, -N02, -NH2, -SH, -CN, halogen, -C(=0)H, -C(=0)CH3, -C02H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C2-C6 alkenyl, and substituted or unsubstituted C2-C6 alkynyl, said C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, and C2-C6 alkynyl are optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6 alkyl; R c5 , R c6 , R c7 , and R c8 are each independently selected from the group consisting of R c0 ; preferably, one and only one of R c5 , R c6 , R c7 , and R c8 is R c0 ; R c9 R c0 , and R c9 is not R c0 For R ca -(R cb ) m0 -, R ca selected from R cb each occurrence is independently selected from a single bond, methylene, -C(O)-, -N(R0)-, m0is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; m1is at each occurrence independently selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, and 7; m2is at each occurrence independently selected from the group consisting of 1, 2, 3, 4, 5, and 6; R3 is CR 3a , R4 is CR 4a , and R3 and R4 form, with the atoms to which they are attached, a Time: when X1is -CN, R c1 is not when X1is -OH, R c1 is not Preferably, when R3 is CR 3a , R4 is CR 4a , and R3 and R4 form, with the atoms to which they are attached, a at the time, R c1 not for X2, X3, X4, X5, and X6are at each occurrence independently selected from the group consisting of H, OR0, SR0, SOR0, SO2R0, C(=0)R0, C(=0)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, C1-C6alkoxy, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; preferably, X2, X3, X4, X5, and X6are at each occurrence independently selected from the group consisting of H, OR0, SR0, SOR0, SO2R0, C(=0)R0, C(=0)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; R0and R 00 each occurrence is independently selected from the group consisting of H, OH, N02, NH2, SH, CN, halogen, -C(=0)H, -C(=0)-Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; preferably, R0and R 00 each occurrence is independently selected from the group consisting of H, OH, N02, NH2, SH, CN, halogen, -C(=0)H, -C(=0)-Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; preferably, R0and R 1-6 alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said Ci-C6alkyl, Ci-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and Ci-C6alkyl; more preferably, R0and R 00 each occurrence is independently selected from the group consisting of H, OH, N02, NH2, SH, CN, halogen, -C(=0)H, -C(=0)-Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; preferably, R0and R 1-3 alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said Ci-C6alkyl, Ci-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and Ci-C6alkyl; more preferably, R0and R 00 each occurrence is independently selected from the group consisting of H, OH, F, Cl, Br, I, substituted or unsubstituted Ci-C3alkoxy, and substituted or unsubstituted Ci-C3alkyl, said Ci-C3alkyl, and Ci-C3alkoxy being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and Ci-C3alkyl; Cy is selected from the group consisting of 3-10 membered cycloalkylene, 3-10 membered heterocyclylene containing at least one heteroatom independently selected from the group consisting of N, O, and S, 6-10 membered arylene, and 5-10 membered heteroarylene containing at least one heteroatom independently selected from the group consisting of N, O, and S, the 3-10 membered cycloalkylene, 3-10 membered heterocyclylene, 6-10 membered arylene, and 5-10 membered heteroarylene optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, C 1-6 alkyl, C 1-6 alkoxy, hydroxyl, -NO2, -NH2, and -CN; preferably, Cy is phenylene, the phenylene optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, C 1-6 alkyl, C 1-6 alkoxy, hydroxyl, -NO2, -NH2, and -CN; and n is at each occurrence independently selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; Preferably, R3 is CR 3a , R4 is CR 4a , and when R3 and R4 form a ring with the atoms to which they are attached, the ring structure formed is selected from: Preferably, when R3is not linked with R4to form a ring, R 3b is The compound of claim 1 or 2, wherein R1is selected from the group consisting of H, F, Cl, Br, I, hydroxyl, -OC(=0)0-Ci-C6alkyl, and Ci-C6alkyl; preferably, R1is selected from the group consisting of H and -OC(=0)0-Ci-C3alkyl; preferably, R1is -OC(=0)CH3; and / or, R2is selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and Ci-C6alkyl; preferably, R2is selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and Ci-C3alkyl; preferably, R2is methyl; and / or Y1is hydroxyl; and / or Y2is selected from the group consisting of H, Ci-C6alkyl, and Ci-C6alkoxy; preferably, Y2is selected from the group consisting of H, Ci-C3alkyl, and Ci-C3alkoxy; preferably, Y2is Ci-C3alkoxy; preferably, Y2is -OCH3; and / or Y3is selected from the group consisting of H, Ci-C6alkyl, and Ci-C6alkoxy; preferably, Y3is selected from the group consisting of H, Ci-C3alkyl, and Ci-C3alkoxy; preferably, Y3is Ci-C3alkyl; preferably, Y3is CH3; and / or Y4, Y5, Y6, Y8, and Y9are each independently selected from the group consisting of H, Ci-C3alkyl, and Ci-C3alkoxy; preferably, Y4, Y5, Y6, Y8, and Y9are each independently H; and / or Y7is selected from the group consisting of H, Ci-C6alkyl, and Ci-C6alkoxy; preferably, Y7is selected from the group consisting of H, Ci-C3alkyl, and Ci-C3alkoxy; preferably, Y7is selected from the group consisting of H, and Ci-C3alkyl; Y7is Ci-C3alkyl; preferably, Y7is CH3; and / or X1is at each occurrence independently selected from the group consisting of halogen, Ci-C6alkyl, Ci-C6haloalkyl, hydroxyl, Ci-C6alkoxy, CN, (=0), and H; preferably, X1is at each occurrence independently selected from the group consisting of F, Cl, Br, I, Ci-C3alkyl, Ci-C3haloalkyl, hydroxyl, Ci-C3alkoxy, CN, and H; preferably, X1is selected from the group consisting of hydroxyl and CN; and / or R3is CR 3a , R4is CR 4a ; preferably, R3and R4are each independently selected from CH, C-F, C-Cl, C-Br, C-I, C-hydroxyl, and C-C1-C3alkyl; preferably, R3and R4are each independently CH; and R3and R4together with the atoms to which they are attached form and / or R a1 and R a2 are each independently selected from the group consisting of H, F, CI, Br, I, hydroxyl, and C1-C6 alkyl; preferably, R a1 and R a2 are each independently selected from the group consisting of H, F, CI, Br, I, hydroxyl, and C1-C3 alkyl; preferably, R a1 and R a2 are each independently H; and / or R5and R6are each independently selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and Ci-C6alkyl; preferably, R5and R6are each independently selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and Ci-C3alkyl; preferably, R5and R6are each independently H; and / or Q4is CR0or N, Q3is CR0or N; and at least one of Q3and Q4is N; more preferably, Q4is CH or N, Q3is CH or N, at least one of Q3and Q4is N; and / or Q5is NH; and / or Q6is C; and / or Q2is selected from the group consisting of O, S, C(Ci-C6alkyl)2, C(Ci-C6alkyl)H, CH2, N(Ci-C6alkyl), and NH; preferably, Q2is selected from the group consisting of O, S, C(Ci-C3alkyl)2, C(Ci-C3alkyl)H, CH2, N(Ci-C3alkyl), and NH; preferably, Q2is selected from the group consisting of O, CH2, and NH; and / or Q2is selected from the group consisting of O, S, C(Ci-C6alkyl)2, C(Ci-C6alkyl)H, CH2, N(Ci-C6alkyl), and NH; preferably, Q2is selected from the group consisting of O, S, C(Ci-C3alkyl)2, C(Ci-C3alkyl)H, CH2, N(Ci-C3alkyl), and NH; preferably, Q2is selected from the group consisting of O, CH2, and NH; and / or Q1is selected from O, S, C(Ci-C6alkyl)2, C(Ci-C6alkyl)H, CH2, N(Ci-C6alkyl), and NH; preferably, Q1is selected from O, S, C(Ci-C3alkyl)2, C(Ci-C3alkyl)H, CH2, N(Ci-C3alkyl), and NH; preferably, Q1is selected from O, CH2, and NH; preferably, Q1is selected from O and NH; R c1 , R c2 , R c3 , R c4 , R c6 , R c7 , and R c8 are each independently selected from R c0 , H, OH, -N02, -NH2, -SH, -CN, halogen, -C(=0)H, -C(=0)CH3, -C02H, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxyl, substituted or unsubstituted C2-C6alkenyl, and substituted or unsubstituted C2-C6alkynyl, said Ci-C6alkyl, Ci-C6alkoxyl, C2-C6alkenyl, and C2-C6alkynyl optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and Ci-C6alkyl; R is selected from H, F, CI, Br, I, hydroxyl, and C1-C6 alkyl; preferably, R c6 R is selected from H, F, CI, Br, I, hydroxyl, and C1-C6 alkyl; preferably, R c6 R is selected from H, F, CI, Br, I, hydroxyl, and C1-C6 alkyl; preferably, R c6 R is selected from H and F; and / or R is selected from H, F, CI, Br, I, hydroxyl, and Ci-C6alkyl; preferably, R c7 R is selected from H, F, CI, Br, I, hydroxyl, and Ci-C6alkyl; preferably, R c7 R is selected from H, F, CI, Br, I, hydroxyl, and Ci-C6alkyl; preferably, R c7 R is H; and / or R is selected from H, F, CI, Br, I, hydroxyl, and Ci-C6alkyl; preferably, R c8 R is selected from H, F, CI, Br, I, hydroxyl, and Ci-C6alkyl; preferably, R c8 R is selected from H, F, CI, Br, I, hydroxyl, and Ci-C6alkyl; preferably, R c8 R is H; and / or R c5 selected from R c0 , -NH2, and -(CH2) 1-4 -OH; R c0 For R ca -(R cb ) m0 -, R ca selected from R cb each occurrence is independently selected from methylene, -C(O)-, -O-, -N(R0)-, Preferably, R cb Each occurrence is independently selected from methylene, -C(O)-, -N(R0)-, m0is selected from 0, 1, 2, 3, 4, 5, and 6; m1is at each occurrence independently selected from 0, 1, and 2; preferably, m1is at each occurrence independently 1; m2is at each occurrence independently selected from 1, and 2; preferably, m2is at each occurrence independently 1; Preferably, R c5 Selected from -NH2 and -(CH2) m3 OH, -(CH2) m3 NHCH3、-(CH2) m3 NH2, m1is at each occurrence independently selected from 0, 1, and 2; preferably, m1is at each occurrence independently 1; m2is at each occurrence independently selected from 1, and 2; preferably, m2is at each occurrence independently 1; m3is selected from 0, 1, 2, 3, 4, and 5; m4is selected from 0, 1, 2, 3, 4, and 5; preferably, m4is selected from 0, and 1; Preferably, R c5 selected from -NH2, -(CH2) 1-4 -OH, and / or Preferably, R c9 selected from and / or X2, X3, X4, X5, and X6are at each occurrence independently selected from H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said Ci-C6alkyl, Ci-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl are optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and Ci-C6alkyl; preferably, X2, X3, X4, and X5are at each occurrence independently selected from H, F, Cl, Br, I, hydroxyl, and Ci-C6alkyl; preferably, X2, X3, X4, and X5are at each occurrence independently selected from H, F, Cl, Br, I, hydroxyl, and Ci-C3alkyl; preferably, X2, X3, X4, and X5are at each occurrence independently H; and / or preferably, X6is selected from H, F, Cl, Br, I, hydroxyl, Ci-C6alkyl, -Ci-C6alkyl-OH, and -Ci-C6alkyl-NH2; preferably, X6is selected from H, F, Cl, Br, I, hydroxyl, Ci-C3alkyl, -Ci-C3alkyl-OH, and -Ci-C3alkyl-NH2; X6is selected from H, -CH2-OH, and -CH2-NH2; and / or R0and R 00 each occurrence is independently selected from the group consisting of H, OH, N02, NH2, SH, CN, F, CI, Br, I, -C(=0)H, -C(=0)-C 1-6 alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; more preferably, R0and R 00 each occurrence is independently selected from the group consisting of H, OH, N02, NH2, SH, CN, F, CI, Br, I, -C(=0)H, -C(=0)-C 1-3 alkyl, substituted or unsubstituted C1-C3alkoxy, substituted or unsubstituted C1-C3alkyl, substituted or unsubstituted C2-C3alkenyl, substituted or unsubstituted C2-C3alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said C1-C3alkyl, C1-C3alkoxy, C2-C3alkenyl, C2-C3alkynyl, aryl, and heteroaryl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C3alkyl; more preferably, R0and R 00 each occurrence is independently selected from the group consisting of H, OH, F, CI, Br, I, substituted or unsubstituted C1-C3alkoxy, and substituted or unsubstituted C1-C3alkyl, said C1-C3alkyl, and C1-C3alkoxy being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C3alkyl; more preferably, R0each occurrence is independently selected from the group consisting of H and C1-C3alkyl; more preferably, R0each occurrence is independently selected from the group consisting of H and CH3; and / or Cy is phenylene, which is optionally substituted with a substituent selected from the group consisting of deuterium atom, halogen, C 1-6 alkyl, C 1-6 alkoxy, hydroxy, -NO2, -NH2, and -CN. The compound of any one of claims 1-3, wherein R3 is CR 3a , R4 is CR 4a , and when R3 and R4 form a ring with the atoms to which they are attached, the ring formed is selected from: When R3 is not connected to R4 to form a loop, R 3b for Preferably, R 3a and R 4a are each independently selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and C1-C3 alkyl; preferably, R 3a and R 4a are each independently H. The compound of any one of claims 1-3, or an isomer, isotopic derivative, polymorph, prodrug, pharmaceutically acceptable salt, or solvate thereof, wherein the compound is a1is selected from 0, and 1; wherein X1is at each occurrence independently selected from CN and OH; R c5 selected from -NH2, -(CH2) m3 OH, -(CH2) m3 NHCH3, -(CH2) m3 NH2 m1is at each occurrence independently selected from 0, 1, 2, 3, 4, and 5; preferably, m1is at each occurrence independently selected from 0, 1, 2, and 3; m2is each independently at each occurrence selected from 1, 2, 3, 4, and 5; preferably, m2is each independently at each occurrence selected from 1, 2, and 3; m3is each independently at each occurrence selected from 0, 1, 2, 3, 4, and 5; m4is each independently at each occurrence selected from 0, 1, 2, 3, 4, and 5; preferably, m4is each independently at each occurrence selected from 0 and 1; R c6 each occurrence is independently selected from the group consisting of H, OH, F, Cl, Br, I, and C1-C3 alkyl; preferably, R c6 each occurrence is independently selected from the group consisting of H and F; Q1and Q2are each independently at each occurrence selected from NR0, O, S, and C(R0)2; preferably, Q1is selected from NR0, O, and S; Q2is selected from C(R0)2, O, and S; preferably, Q1is selected from NH; Q2is selected from CH2, and O; R0is each independently at each occurrence selected from H, OH, F, Cl, Br, I, C1-C3alkoxy, and C1-C3alkyl; preferably, R0is each independently at each occurrence selected from H and C1-C3alkyl; more preferably, R0is each independently at each occurrence selected from H and CH3; X6is selected from the group consisting of H, -(CH2) m1 OH, and -(CH2) m1 NH2; preferably, X6is selected from the group consisting of H, -CH2-OH, and -CH2-NH2; and When R c5 -(CH2) m3 When NHCH3, -NH2, or -NHCOCH2OH, R c6 Selected from OH, F, Cl, Br, I, and C1-C3 alkyl groups; preferably, R c6 It is F. The compound of claim 1 selected from: a drug-linker conjugate as represented by Formula II, or an isomer, isotopic derivative, polymorph, prodrug, pharmaceutically acceptable salt, or solvate thereof, L0-D0(Formula II); wherein, L0is a linker precursor; represents the site of attachment of the terminal group to the rest of the compound; D0is a compound according to any one of claims 1 to 6, wherein the hydrogen atom of the hydroxyl group or the amino group at the end of R 3b , R a1 , R a2 , R c0 , R c1 , R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , Q1, or Q2 is lost to form a group; wherein R 3b , R a1 , R a2 , R c0 , R c1 , R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , Q1, or Q2 is as defined in any one of claims 1 to 6; Preferably, D0is a group formed by the loss of a hydrogen atom from a hydroxyl or amino group at the terminal group of R 3b , R a1 , R a2 , R c0 , R c1 , R c2 , R c3 , R c4 , R c5 , Q1, or Q2; preferably, D0is a group formed by the loss of a hydrogen atom from a hydroxyl or amino group at the terminal group of R 3b , R a1 , R a2 , R c0 , Q1, or Q2; preferably, D0is a group formed by the loss of a hydrogen atom from a hydroxyl or amino group at the terminal group of R c5 , Q1, or Q2; the terminal fragment of the group after the loss of the hydrogen atom is selected from: wherein represents the site of attachment of D0to L0after loss of a hydrogen atom; R0and m1are as defined in any one of claims 1-6; preferably, m1is 1; preferably, R0is selected from -CH3and H; the linker precursor forms a linker upon attachment to the antibody. a drug-linker conjugate as represented by Formula II-1, or an isomer, isotopic derivative, polymorph, prodrug, pharmaceutically acceptable salt, or solvate thereof, L0-D0(Formula II-1); wherein, L0is a linker precursor; wherein R1and R2are each independently selected from H, halogen, hydroxyl, C(=O)R0, C(=O)OR0, -OC(=O)OR0, -OC(=O)R0, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted C1-C6alkoxy, and substituted or unsubstituted C1-C6haloalkyl; preferably, R1and R2are each independently selected from H, halogen, hydroxyl, C(=O)R0, C(=O)OR0, -OC(=O)OR0, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted C1-C6alkoxy, and substituted or unsubstituted C1-C6haloalkyl, said C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C1-C6alkoxy, and C1-C6haloalkyl are optionally substituted with a substituent selected from a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, and Y9are each independently selected from H, C1-C6alkyl, C1-C6alkoxy, C1-C6haloalkyl, OH, NO2, NH2, SH, CN, or halogen; D0 is as shown in formula I-2, ​ ​ X1is independently at each occurrence selected from the group consisting of halogen, alkyl, haloalkyl, OR x , CN, (=0) and H; wherein R x is selected from the group consisting of H, alkyl, haloalkyl, alkenyl, alkynyl, OH, OR0, SR0, SOR0and SO2R0; R3and R4are each independently selected from CH, C-F, C-Cl, C-Br, C-I, C-hydroxyl, and C-Ci-C6alkyl; and R3and R4, together with the atoms to which they are attached, form Q1is independently selected at each occurrence from NR 00 , C(R 00 )2, O, and S; Q2is each independently for each occurrence selected from the group consisting of a single bond, NR 00 , C(R 00 )2, O, and S; R5is selected from the group consisting of H, halogen, C(=O)R0, C(=O)OR0, substituted or unsubstituted C1-C6alkyl, C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl; preferably, R5is selected from the group consisting of H, halogen, C(=O)R0, C(=O)OR0, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, and substituted or unsubstituted C2-C6alkynyl, said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, and C2-C6alkynyl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; Q4is CR0or N, Q3is CR0or N; Q5is NR0; X2, X3, X4, X5, and X6are each independently at each occurrence selected from the group consisting of H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, C1-C6alkoxy, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; preferably, X2, X3, X4, X5, and X6are each independently at each occurrence selected from the group consisting of H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; R0and R 00 each occurrence is independently selected from the group consisting of H, OH, N02, NH2, SH, CN, halogen, -C(=0)H, -C(=0)-Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; preferably, R0and R 00 each occurrence is independently selected from the group consisting of H, OH, N02, NH2, SH, CN, halogen, -C(=0)H, -C(=0)-Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; preferably, R0and R 1-6 alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said Ci-C6alkyl, Ci-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and Ci-C6alkyl; more preferably, R0and R 00 each occurrence is independently selected from the group consisting of H, OH, N02, NH2, SH, CN, halogen, -C(=0)H, -C(=0)-Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; preferably, R0and R 1-3 alkyl, substituted or unsubstituted Ci-C3alkoxy, substituted or unsubstituted Ci-C3alkyl, substituted or unsubstituted C2-C3alkenyl, substituted or unsubstituted C2-C3alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said Ci-C3alkyl, Ci-C3alkoxy, C2-C3alkenyl, C2-C3alkynyl, aryl, and heteroaryl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and Ci-C3alkyl; more preferably, R0and R 00 each occurrence is independently selected from the group consisting of H, OH, F, Cl, Br, I, substituted or unsubstituted Ci-C3alkoxy, and substituted or unsubstituted Ci-C3alkyl, said Ci-C3alkyl, and Ci-C3alkoxy being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and Ci-C3alkyl; n1is selected from the group consisting of 0, 1, 2, 3, 4; La is selected from the group of 1, 2, 3, or a combination of more of a single bond, -O-, C2-C6alkenylene, C2-C6alkynylene, C1-C6alkylene, C1-C6haloalkylene, -NR 00 , -C(O)-, -C(S)-, -SO2-, C3-C6cyclylene, C3-C6heterocyclylene containing 1, 2, or 3 heteroatoms independently selected from N, O, and S; preferably, La is selected from the group of 1, 2, 3, 4, 5, 6, 7, 8, or 9 of a single bond, -O-, C2-C3alkynylene, C1-C3alkylene, C1-C3haloalkylene, -NH-, -NCH3-, -C(O)-, -C(S)-, -SO2-, C3-C6cyclylene, and C3-C6heterocyclylene containing 1, 2, or 3 heteroatoms independently selected from N, O, and S; preferably, La is selected from the group of 1, 2, 3, 4, 5, 6, 7, 8, or 9 of a single bond, -O-, -(CH2) m3 , -(CH2) m3 , -(CH2) m3 , -(CH2) m1is each independently at each occurrence selected from the group consisting of 0, 1, and 2; preferably, m1is each independently at each occurrence 1; m2is each independently at each occurrence selected from the group consisting of 1 and 2; preferably, m2is each independently at each occurrence 1; m3is each independently at each occurrence selected from the group consisting of 0, 1, 2, 3, 4, and 5; m4is each independently at each occurrence selected from the group consisting of 0, 1, 2, 3, 4, and 5; preferably, m4is selected from the group consisting of 0 and 1; L0 is L j -L p -L 2 -L 3 -L 4 -L 5 -; wherein, L j for wherein X S is a single bond or N(C 1-6 alkyl); each R is independently a thiol-reactable group; L p is selected from a single bond, a 6-10 membered arylene group, and a 5-8 membered heteroarylene group comprising 1 to 3 heteroatoms independently selected from N, O, and S as ring atoms, wherein the 6-10 membered arylene group and the 5-8 membered heteroarylene group are each independently optionally substituted with one or more R 1L substituents; L 2 is a single bond, or a combination of one or more selected from the group consisting of alkynylene, alkenylene, NR 2L , O, C(O), alkylene, and heteroalkylene, wherein the alkynylene, alkenylene, alkylene, and heteroalkylene are optionally substituted with one or more R 2L ; L 3 for wherein r1 is selected from the group consisting of natural numbers from 1 to 20; r2 is selected from the group consisting of natural numbers from 0 to 20; r3 is selected from the group consisting of 1, 2, 3; r4 is selected from the group consisting of 0, 1, 2, 3, 4, 5; r5 is selected from the group consisting of integers from 1 to 30, wherein L2 represents the attachment site to L 2 represents the attachment site to L L4 represents the attachment site to L 4 represents the attachment site to L ; preferably, r1 is selected from the group consisting of 1, 2, 3, 4, 5, 6; r2 is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6; r4 is selected from the group consisting of 0, 1, 2; r5 is selected from the group consisting of integers from 5 to 15, preferably r5 is selected from the group consisting of 10, 1 1, 12, 13, 14, 15, Ar 1 each occurrence is independently selected from triazidylene, phenylene, pyrrolidinylene, piperidinylene, cyclohexanylene, thiazolylene, and dihydro-pyridazinylene, optionally substituted with one or more substituents selected from hydroxy, cyano, amino, C 1-6 alkyl, haloC 1-6 alkyl, C 1-6 alkoxy, haloC 1-6 alkoxy and C 3-8 cycloalkyl; Preferably, Ar 1 each occurrence is selected independently from triazolene, phenylene, pyrrolidinylene, piperidinylene, cyclohexanylene, thiazolene, and dihydropyridazinylene, said triazolene, phenylene, pyrrolidinylene, piperidinylene, cyclohexanylene, thiazolene, and dihydropyridazinylene being optionally substituted with one or more substituents selected from the group consisting of hydroxy, cyano, amino, and C1-C4-alkyl; 1-6 substituted with one or more substituents selected from the group consisting of hydroxy, cyano, amino, and C1-C4-alkyl; Preferably, Ar 1 each occurrence is selected independently from the group consisting of Preferably, Ar 1 each occurrence independently is L 4 peptide containing 2 to 7 amino acid residues, wherein the amino acids are optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, amino, alkyl, haloalkyl, deuterium- substituted alkyl, alkoxy, haloalkoxy, and cycloalkyl; L 5 selected from a single bond, L4 -NR 2L (CR 3L R 4L ) t -Z1-(CR 3L R 4L ) t -C(=O)-* D0 , L4 -NR 2L (CR 3L R 4L ) t - D0 , L4 -NR 2L (CR 3L R 4L ) t -Z1-(CR 3L R 4L ) t -Z1-C(=O)-* D0 , L4 -NR 2L -(CR 3L R 4L ) t -Z1-Ar 3 -C(=O)-* D0 , L4 -NR 2L -Ar 2 -(CR 3L R 4L ) t -Z1-C(=O)-NR 2L -(CR 3L R 4L ) t -Ar 2 -NR 2L -C(=O)-* D0 and L4 -NR 2L -Ar 2 -(CR 3L R 4L ) t -Z1-C(=O)-* D0 ; L4 denotes the point of attachment to L 4 , and D0 denotes the point of attachment to D0; Wherein, t is independently selected from 0, 1, 2, 3, 4, 5, and 6 each time it appears; Z1 is independently selected as a single bond, O, S, or NH each time it appears; Ar 2 Each occurrence is independently arylene or heteroarylene, the heteroarylene containing one, two, or three heteroatoms independently selected from N, O, and S as ring atoms, wherein the arylene or heteroarylene is optionally surrounded by one or more atoms selected from hydrogen, halogen, hydroxyl, cyano, C 1-6 Alkyl, -OC 1-6 Alkyl, amino, -NH(C) 1-6 alkyl) and -N(C) 1-6 Substitution of alkyl group 2; Ar 3 independently at each occurrence is cycloalkylene or heterocycloalkylene containing 1, 2, or 3 heteroatoms independently selected from N, O, and S as ring atoms, wherein the cycloalkylene or heterocycloalkylene is optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxyl, cyano, C 1-6 alkyl, -O-C 1-6 alkyl, amino, -NH(C 1-6 alkyl), and -N(C 1-6 alkyl)2; R 1L , R 2L , R 3L , R 4L and R 5L are each independently at each occurrence selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 alkoxy, haloC 1-6 alkyl, haloC 1-6 alkyl, deuterated C 1-6 alkyl, carboxyl, -C 1-6 alkylene-NH2, and -C 1-6 alkylene-OH; represents a point of attachment. The drug linker conjugate of claim 8, wherein: R1is selected from the group consisting of H, F, Cl, Br, I, hydroxyl, -OC(=O)O- C1-C6alkyl, and C1-C6alkyl; preferably, R1is selected from the group consisting of H and -OC(=O)O-C1-C3alkyl; preferably, R1is -OC(=O)CH3; and / or R2is selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and C1-C6alkyl; preferably, R2is selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and C1-C3alkyl; preferably, R2is methyl; and / or Y1, Y2, Y3, Y4, Y5, Y6, Y8, and Y9are each independently selected from the group consisting of H, C1-C3alkyl, C1-C3alkoxy, C1-C3haloalkyl, OH, NO2, NH2, SH, CN, or halogen; preferably, Y1is OH; and / or Y2is -OCH3; and / or Y3is CH3; and / or Y4, Y5, Y6, Y8, and Y9are each independently H; and / or Y7is selected from the group consisting of H, C1-C6alkyl, and C1-C6alkoxy; preferably, Y7is selected from the group consisting of H, C1-C3alkyl, and C1-C3alkoxy; preferably, Y7is selected from the group consisting of H, and C1-C3alkyl; Y7is C1-C3alkyl; preferably, Y7is CH3; X1is at each occurrence independently selected from the group consisting of halogen, C1-C6alkyl, C1-C6haloalkyl, hydroxyl, C1-C6alkoxy, CN, (=0), and H; preferably, X1is at each occurrence independently selected from the group consisting of F, Cl, Br, I, C1-C3alkyl, C1-C3haloalkyl, hydroxyl, C1-C3alkoxy, CN, and H; preferably, X1is selected from the group consisting of hydroxyl and CN; Q1is selected from the group consisting of O, S, C(C1-C6alkyl)2, C(C1-C6alkyl)H, CH2, N(C1-C6alkyl), and NH; preferably, Q1is selected from the group consisting of O, S, C(C1-C3alkyl)2, C(C1-C3alkyl)H, CH2, N(C1-C3alkyl), and NH; preferably, Q1is selected from the group consisting of O, CH2, and NH; preferably, Q1is selected from the group consisting of O and NH; and / or Q2is selected from the group consisting of O, S, C(C1-C6alkyl)2, C(C1-C6alkyl)H, CH2, N(C1-C6alkyl), and NH; preferably, Q2is selected from the group consisting of O, S, C(C1-C3alkyl)2, C(C1-C3alkyl)H, CH2, N(C1-C3alkyl), and NH; preferably, Q2is selected from the group consisting of O, CH2, and NH; and / or Q4is CH or N, Q3is CH or N, at least one of Q3and Q4is N; and / or Q5is NH; and / or R5is selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and C1-C6alkyl; preferably, R5is selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and C1-C3alkyl; preferably, R5is H; and / or R3and R4are each independently selected from CH, C-F, C-Cl, C-Br, C-I, C-hydroxyl, and C-Ci-C3alkyl; preferably, R3and R4are each independently CH; and R3and R4together with the atoms to which they are attached form formula (II-A), formula (II-B), formula (II-C), or (II-F), preferably, R3and R4together with the atoms to which they are attached form Preferably, R3and R4together with the atoms to which they are attached form Preferably, the ring structure formed by R3and R4is selected from: X2, X3, X4, and X5are at each occurrence independently selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and C1-C6alkyl; preferably, X2, X3, X4, and X5are at each occurrence independently selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and C1-C3alkyl; preferably, X2, X3, X4, and X5are at each occurrence independently H; and / or X6is selected from the group consisting of H, F, CI, Br, I, hydroxyl, C1-C6alkyl, -C1-C6alkyl-OH, and -C1-C6alkyl-NH2; preferably, X6is selected from the group consisting of H, F, CI, Br, I, hydroxyl, C1-C3alkyl, -C1-C3alkyl-OH, and -C1-C3alkyl-NH2; X6is selected from the group consisting of H, -CH2-OH, and -CH2-NH2; and / or R0is each independently at each occurrence selected from the group consisting of H and C1-C3alkyl; more preferably, R0is each independently at each occurrence selected from the group consisting of H and CH3; and / or R 00 each occurrence is independently selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and C1-C6 alkyl; preferably, R 00 each occurrence is independently selected from the group consisting of H, F, Cl, Br, I, hydroxyl, and C1-C3 alkyl; and / or n1is selected from the group consisting of 0, and 1 ; and / or Preferably, La is selected from a single bond, The drug linker conjugate of claim 8 or 9, wherein: L j for wherein X S is selected from a single bond, NH and N-CH3; preferably, X S is a single bond; Preferably, L j selected from each R is independently selected from the group consisting of halogen and -S-Ar 4 ; wherein Ar 4 is selected from the group consisting of phenyl, C 1-6 alkylphenyl-, C 1-6 alkoxyphenyl-, 2-pyridyl, 2-pyrimidinyl, 1 -methylimidazol-2-yl, said phenyl, C 1-6 alkylphenyl-, C 1-6 alkoxyphenyl-, 2-pyridyl, 2-pyrimidinyl and 1-methylimidazol-2-yl groups are optionally substituted with 1, 2, 3 or 4 R a substituents; wherein W2is selected from the group consisting of amino, -NR a -C 1-6 alkyl, -NR a -C 1-6 alkoxy, -NR a -C 1-6 alkylene-NR b R c , -NR a -C 1-6 alkylene-C 1-6 alkoxy and 5- to 8-membered heterocycloalkyl, which contains 1, 2, 3 or 4 heteroatoms selected from the group consisting of N, O and S as ring members, wherein the S atom is optionally substituted with 1 or 2 oxo groups; preferably, W2is selected from the group consisting of amino, -NR a -C 1-6 alkyl, R a each occurrence is selected independently from the group consisting of halogen, C 1-6 alkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, deuterated C 1-6 alkyl, carboxyl, -C 1-6 alkylene-C 1-6 alkoxy, -C 1-6 alkylene-NH2and -C 1-6 alkylene-OH; R b , R c each occurrence is independently selected from C 1-6 alkyl and C 1-6 alkoxy; Preferably, each occurrence of R is independently selected from halogens, More preferably, each R is independently selected from chloro, bromo, Preferably, L j selected from The drug linker conjugate of any one of claims 8-10, wherein, L p is selected from a single bond, phenylene, and 5-8 membered heteroarylene; said 5-8 membered heteroarylene comprises 1 to 3 heteroatoms independently selected from N, O, and S as ring atoms, wherein said phenylene and 5-8 membered heteroarylene are each independently optionally substituted with one or more R 5L substituents; Preferably, each of said phenylene and 5-8 membered heteroarylene is independently optionally substituted with 1, 2, 3, 4, 5, or 6 R 5L substituted; Preferably, L p is selected from a single bond, phenylene, and pyridylene; wherein each of said phenylene and pyridylene is independently optionally substituted with 1, 2, 3, or 4 R 5L ; and / or R 5L independently at each occurrence selected from halogen, hydroxyl, cyano, amino, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, haloC 1-6 alkoxy, and C 3-8 cycloalkyl; R is independently at each occurrence selected from halogen, hydroxy, cyano, C 5L R is independently at each occurrence selected from halogen, hydroxy, cyano, C 1-3 R is independently at each occurrence selected from halogen, hydroxy, cyano, C 1-3 R is independently at each occurrence selected from halogen, hydroxy, cyano, C Preferably, R 5L Each time it appears, it is independently selected from Cl, Br, F, -OH, -CN, methyl and -OCH. 3； Preferably, R 5L For F; Preferably, L p is selected from a single bond, phenylene and pyridylene; wherein each of said phenylene and pyridylene is independently optionally substituted with 1, 2, 3 or 4 substituents selected from CI, Br, F, -OH, -CN, methyl and -OCH3; Preferably, L p selected from wherein Lj represents the attachment site to L j represents the attachment site to L L2 represents the attachment site to L 2 represents the attachment site to L The drug linker conjugate of any one of claims 8-11, wherein L 4 is a peptide containing 2, 3, 4, 5 or 6 amino acids, wherein the amino acids are optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxy, cyano, amino, C 1-6 alkyl, halogenated C 1-6 alkyl, deuterated C 1-6 alkyl, C 1-6 alkoxy and C 5-8 cycloalkyl; preferably, L 4 is a peptide comprising 2-6 amino acid residues selected from the group consisting of glycine, phenylalanine, alanine, valine, lysine, citrulline, serine, glutamic acid and aspartic acid; Preferably, L 4 is a dipeptide, tripeptide or tetrapeptide consisting of amino acid residues selected from alanine, phenylalanine, valine, glycine, lysine and citrulline; Preferably, L 4 is a peptide selected from the group consisting of glycine-phenylalanine-glycine, alanine-alanine-alanine-glycine, alanine-alanine-alanine, glycine-glycine-phenylalanine, glycine-glycine-phenylalanine-glycine, valine-citrulline and valine-alanine; wherein, The peptides are optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, amino, alkyl, haloalkyl, deuterium- substituted alkyl, alkoxy, and cycloalkyl, preferably, optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, amino, and C 1-6 alkyl substituents; Preferably, L 4 selected from wherein L3 represents the attachment site to L 3 represents the attachment site to L L5 represents the attachment site to L 5 represents the attachment site to L The drug linker conjugate of any one of claims 8-12, wherein, t is each independently at each occurrence 0, 1, or 2; and / or Z1is each independently at each occurrence a single bond, O, or S; preferably a single bond or O; more preferably O; and / or Ar 2 independently at each occurrence is phenylene optionally substituted with one or more substituents selected from halo, hydroxy, cyano, C 1-6 alkyl, -O-C 1-6 alkyl, amino, -NH(C 1-6 alkyl) and -N(C 1-6 alkyl)2; preferably, Ar 2 independently at each occurrence is Ar 3 independently for each occurrence -4-6 membered heterocycloalkylene optionally substituted with one or more substituents selected from halogen, hydroxyl, cyano, C 1-6 alkyl, -O-C 1-6 alkyl, amino, -NH(C 1-6 alkyl) and -N(C 1-6 alkyl)2; preferably, Ar 3 independently for each occurrence -4-6 membered heterocycloalkylene optionally substituted with one or more substituents selected from halogen, hydroxyl, cyano, C Preferably, L 5 selected from a single bond, and / or L 2 is O; and / or L 3 selected from The drug linker conjugate of any one of claims 8-13, wherein L0is selected from: represents the point of attachment of L0to D0. The drug linker conjugate as claimed in any one of claims 8-14, wherein, D0 is selected from represents the point of attachment of D0to L0. The drug linker conjugate of any one of claims 8-15, selected from: An antibody-drug conjugate of Formula III, or a pharmaceutically acceptable salt thereof, Ab-(L-D0) p (Formula III) wherein, Ab is an antibody or an antigen binding fragment thereof; p is an integer or a decimal number from 0 to 20, but not 0; L is a linker which is -T-L p -L 2 -L 3 -L 4 -L 5 -; L p , L 2 , L 3 , L 4 , L 5 , and D0 are as defined in any one of claims 8-15; T is selected from Preferably, D0 is as shown in formula I-2, wherein R1and R2are each independently selected from the group consisting of H, halogen, hydroxyl, C(=O)R0, C(=O)OR0, -OC(=O)OR0, -OC(=O)R0, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted C1-C6alkoxy, and substituted or unsubstituted C1-C6haloalkyl; preferably, R1and R2are each independently selected from the group consisting of H, halogen, hydroxyl, C(=O)R0, C(=O)OR0, -OC(=O)OR0, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted C1-C6alkoxy, and substituted or unsubstituted C1-C6haloalkyl, said C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C1-C6alkoxy, and C1-C6haloalkyl are optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, and Y9are each independently selected from the group consisting of H, C1-C6alkyl, C1-C6alkoxy, C1-C6haloalkyl, OH, NO2, NH2, SH, CN, or halogen; X1is independently at each occurrence selected from the group consisting of halogen, alkyl, haloalkyl, OR x , CN, (=0) and H; wherein R x is selected from the group consisting of H, alkyl, haloalkyl, alkenyl, alkynyl, OH, OR0, SR0, SOR0and SO2R0; R3and R4are each independently selected from CH, C-F, C-Cl, C-Br, C-I, C-hydroxyl, and C-Ci-C6alkyl; and R3and R4, together with the atoms to which they are attached, form a 3- to 8-membered cycloalkyl ring Q1is independently selected at each occurrence from NR 00 , C(R 00 )2, O, and S; Q2is each independently for each occurrence selected from the group consisting of a single bond, NR 00 , C(R 00 )2, O, and S; R5is selected from the group consisting of H, halogen, C(=O)R0, C(=O)OR0, substituted or unsubstituted C1-C6alkyl, C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl; preferably, R5is selected from the group consisting of H, halogen, C(=O)R0, C(=O)OR0, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, and substituted or unsubstituted C2-C6alkynyl, said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, and C2-C6alkynyl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; Q4is CR0or N, Q3is CR0or N; Q5is NR0; X2, X3, X4, X5, and X6are each independently at each occurrence selected from the group consisting of H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, C1-C6alkoxy, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; preferably, X2, X3, X4, X5, and X6are each independently at each occurrence selected from the group consisting of H, OR0, SR0, SOR0, SO2R0, C(=O)R0, C(=O)OR0, NO2, N(R0)2, NHC(O)R0, CN, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and C1-C6alkyl; R0and R 00 each occurrence is independently selected from the group consisting of H, OH, N02, NH2, SH, CN, halogen, -C(=0)H, -C(=0)-Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; preferably, R0and R 00 each occurrence is independently selected from the group consisting of H, OH, N02, NH2, SH, CN, halogen, -C(=0)H, -C(=0)-Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; preferably, R0and R 1-6 alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said Ci-C6alkyl, Ci-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and Ci-C6alkyl; more preferably, R0and R 00 each occurrence is independently selected from the group consisting of H, OH, N02, NH2, SH, CN, halogen, -C(=0)H, -C(=0)-Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; preferably, R0and R 1-3 alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, said Ci-C6alkyl, Ci-C6alkoxy, C2-C6alkenyl, C2-C6alkynyl, aryl, and heteroaryl being optionally substituted with a substituent selected from the group consisting of a deuterium atom, halogen, OH, NH2, and Ci-C6alkyl; more preferably, R0and R 00 each occurrence is independently selected from the group consisting of H, OH, N02, NH2, SH, CN, halogen, -C(=0)H, -C(=0)-Ci-C6alkyl, substituted or unsubstituted Ci-C6alkoxy, substituted or unsubstituted Ci-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl; preferably, R0and R n1is selected from 0, 1, 2, 3, 4; La is selected from single bond, -O-, C2-C6 alkenyl group, C2-C6 alkyne group, C1-C6 alkylene group, C1-C6 haloalkylene group, -NR group. 00 -, -C(O)-, -C(S)-, -SO2-, C3-C6 cyclic subgroup, C3-C6 heterocyclic subgroup, or a combination of 1, 2, 3, or more thereof, wherein the heterocyclic subgroup contains 1, 2, or 3 heteroatoms independently selected from N, O, and S; preferably, La is selected from single bonds, -O-, C2-C3 ynylene, C1-C3 alkylene, C1-C3 haloalkylene, -NH-, -NCH3-, -C(O)-, -C(S)-, -SO2-, C3-C6 cyclic subgroup, and C3-C6 heterocyclic subgroup, or a combination of 1, 2, 3, 4, 5, 6, 7, 8, or 9 thereof, wherein the heterocyclic subgroup contains 1, 2, or 3 heteroatoms independently selected from N, O, and S; preferably, La is selected from single bonds, -NH-, -(CH2)-, and C3-C6 heterocyclic subgroups. m3 O-, -(CH2) m3 N(CH3)-, -(CH2) m3 NH-, m1is at each occurrence independently selected from 0, 1, and 2; preferably, m1is at each occurrence independently 1; m2is at each occurrence independently selected from 1 and 2; preferably, m2is at each occurrence independently 1; m3is selected from 0, 1, 2, 3, 4, and 5; m4is selected from 0, 1, 2, 3, 4, and 5; preferably, m4is selected from 0 and 1; L p is selected from a single bond, a 6-10 membered arylene group, and a 5-8 membered heteroarylene group comprising 1 to 3 heteroatoms independently selected from N, O, and S as ring atoms, wherein each of said 6-10 membered arylene group and 5-8 membered heteroarylene group is independently optionally substituted with one or more R 1L substituents; L 2 is a single bond, or a combination of one or more selected from the group consisting of alkynylene, alkenylene, NR 2L , O, C(O), alkylene, and heteroalkylene, wherein the alkynylene, alkenylene, alkylene, and heteroalkylene are optionally substituted with one or more R 2L ; L 3 for wherein r1 is selected from the group consisting of natural numbers from 1 to 20; r2 is selected from the group consisting of natural numbers from 0 to 20; r3 is selected from the group consisting of 1, 2, 3; r4 is selected from the group consisting of 0, 1, 2, 3, 4, 5; r5 is selected from the group consisting of integers from 1 to 30, wherein L2 represents the attachment site to L 2 represents the attachment site to L L4 represents the attachment site to L 4 represents the attachment site to L; preferably, r1 is selected from the group consisting of 1, 2, 3, 4, 5, 6; r2 is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6; r4 is selected from the group consisting of 0, 1, 2; r5 is selected from the group consisting of integers from 5 to 15, preferably r5 is selected from the group consisting of 10, 1 1, 12, 13, 14, 15, Ar 1 each occurrence is independently selected from triazidylene, phenylene, pyrrolidinylene, piperidinylene, cyclohexanylene, thiazolylene, and dihydro- pyridazinylene, optionally substituted with one or more substituents selected from hydroxy, cyano, amino, C 1-6 alkyl, haloC 1-6 alkyl, C 1-6 alkoxy, haloC 1-6 alkoxy and C 3-8 cycloalkyl; Preferably, Ar 1 each occurrence is selected independently from triazolene, phenylene, pyrrolidinylene, piperidinylene, cyclohexanylene, thiazolene, and dihydropyridazinylene, said triazolene, phenylene, pyrrolidinylene, piperidinylene, cyclohexanylene, thiazolene, and dihydropyridazinylene being optionally substituted with one or more substituents selected from the group consisting of hydroxy, cyano, amino, and C1-C4-alkyl; 1-6 substituted with one or more substituents selected from the group consisting of hydroxy, cyano, amino, and C1-C4-alkyl; Preferably, Ar 1 each occurrence is selected independently from the group consisting of Preferably, Ar 1 each occurrence independently is L 4 peptides containing 2 to 7 amino acid residues, wherein the amino acids are optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, amino, alkyl, haloalkyl, deuterium- substituted alkyl, alkoxy, haloalkoxy, and cycloalkyl; L 5 selected from a single bond, L4 -NR 2L (CR 3L R 4L ) t -Z1-(CR 3L R 4L ) t -C(=O)-* D0 , L4 -NR 2L (CR 3L R 4L ) t - D0 , L4 -NR 2L (CR 3L R 4L ) t -Z1-(CR 3L R 4L ) t -Z1-C(=O)-* D0 , L4 -NR 2L -(CR 3L R 4L ) t -Z1-Ar 3 -C(=O)-* D0 , L4 -NR 2L -Ar 2 -(CR 3L R 4L ) t -Z1-C(=O)-NR 2L -(CR 3L R 4L ) t -Ar 2 -NR 2L -C(=O)-* D0 and L4 -NR 2L -Ar 2 -(CR 3L R 4L ) t -Z1-C(=O)-* D0 ; L4 represents the point of attachment to L 4 , and D0 represents the point of attachment to D0; Wherein, t is independently selected from 0, 1, 2, 3, 4, 5, and 6 each time it appears; Z1 is independently selected as a single bond, O, S, or NH each time it appears; Ar 2 Each occurrence is independently arylene or heteroarylene, the heteroarylene containing one, two, or three heteroatoms independently selected from N, O, and S as ring atoms, wherein the arylene or heteroarylene is optionally surrounded by one or more atoms selected from hydrogen, halogen, hydroxyl, cyano, C 1-6 Alkyl, -OC 1-6 Alkyl, amino, -NH(C) 1-6 alkyl) and -N(C) 1-6 Substitution of alkyl group 2; Ar 3 independently at each occurrence is cycloalkylene or heterocycloalkylene containing 1, 2, or 3 heteroatoms independently selected from N, O, and S as ring atoms, wherein the cycloalkylene or heterocycloalkylene is optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxyl, cyano, C 1-6 alkyl, -O-C 1-6 alkyl, amino, -NH(C 1-6 alkyl), and -N(C 1-6 alkyl)2; R 1L , R 2L , R 3L , R 4L and R 5L are each independently at each occurrence selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 alkoxy, haloC 1-6 alkyl, haloC 1-6 alkyl, deuterated C 1-6 alkyl, carboxyl, -C 1-6 alkylene-NH2, and -C 1-6 alkylene-OH; T is selected from represents a point of attachment. The antibody-drug conjugate of claim 17, wherein, the linker is selected from: wherein, represents a point of attachment of the linker to D0. The antibody-drug conjugate of claim 17 or 18, which is selected from the group consisting of: p is selected from an integer or a decimal number from 1 to 12; preferably, p is selected from an integer or a decimal number from 1 to 10; preferably, p is selected from an integer or a decimal number from 2 to 8; preferably, p is selected from an integer or a decimal number from 2 to 6; preferably p is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. The antibody-drug conjugate of any one of claims 17-19, wherein the antibody is selected from the group consisting of a chimeric antibody, a humanized antibody, and a fully human antibody. Preferably, the antibody or antigen-binding fragment thereof is selected from the group consisting of an anti-TROP-2 antibody, an anti-HER2 (ErbB2) antibody, an anti-NECTIN4 antibody, an anti-EGFR antibody, an anti-B7-H3 antibody, an anti-c-Met antibody, an anti-HER3 (ErbB3) antibody, an anti-HER4 (ErbB4) antibody, an anti-LIV-1 antibody, an anti-ROR1 antibody, an anti-CD20 antibody, an anti-CD22 antibody, an anti-CD30 antibody, an anti-CD33 antibody, an anti-CD44 antibody, an anti-CD56 antibody, an anti-CD70 antibody, an anti-CD73 antibody, an anti-CD105 antibody, an anti-CEA antibody, an anti-A33 antibody, an anti-Cripto antibody, an anti-EphA2 antibody, an anti-G250 antibody, an anti-MUC1 antibody, an anti-Lewis Y antibody, an anti-VEGFR antibody, an anti-GPNMB antibody, an anti-Integrin antibody, an anti-PSMA antibody, an anti-Tenascin-C antibody, an anti-SLC44A4 antibody, an anti-Mesothelin antibody, an anti-CDH17 antibody, and an anti-GPC3 antibody or antigen-binding fragment thereof; 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, an anti-CDH17 antibody, and an anti-GPC3 antibody or 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, an anti-CDH17 antibody, and an anti-GPC3 antibody or antigen-binding fragment thereof Preferably, the antibody or antigen-binding fragment thereof is an anti-NECTIN4 antibody or antigen-binding fragment thereof; preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein, The heavy chain comprises a heavy chain variable region comprising 3 heavy chain complementarity determining regions (HCDRs), wherein the amino acid sequence of the heavy chain complementarity determining region 1 (HCDR1) is set forth in SEQ ID NO: 11, the amino acid sequence of the heavy chain complementarity determining region 2 (HCDR2) is set forth in SEQ ID NO: 12, and the amino acid sequence of the heavy chain complementarity determining region 3 (HCDR3) is set forth in SEQ ID NO: 13; and / or the light chain comprises a light chain variable region comprising 3 light chain complementarity determining regions (LCDRs), wherein the amino acid sequence of the light chain complementarity determining region 1 (LCDR1) is set forth in SEQ ID NO: 14, the amino acid sequence of the light chain complementarity determining region 2 (LCDR2) is set forth in SEQ ID NO: 15, and the amino acid sequence of the light chain complementarity determining region 3 (LCDR3) is set forth in SEQ ID NO: 16; preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and / or a light chain variable region, wherein the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 18; and / or the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 17; preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is set forth in SEQ ID NO: 1, and / or the amino acid sequence of the light chain is set forth in SEQ ID NO: 2; The heavy chain comprises a heavy chain variable region comprising 3 heavy chain complementarity determining regions (HCDRs), wherein the amino acid sequence of the heavy chain complementarity determining region 1 (HCDR1) is set forth in SEQ ID NO: 11, the amino acid sequence of the heavy chain complementarity determining region 2 (HCDR2) is set forth in SEQ ID NO: 12, and the amino acid sequence of the heavy chain complementarity determining region 3 (HCDR3) is set forth in SEQ ID NO: 13; and / or the light chain comprises a light chain variable region comprising 3 light chain complementarity determining regions (LCDRs), wherein the amino acid sequence of the light chain complementarity determining region 1 (LCDR1) is set forth in SEQ ID NO: 14, the amino acid sequence of the light chain complementarity determining region 2 (LCDR2) is set forth in SEQ ID NO: 15, and the amino acid sequence of the light chain complementarity determining region 3 (LCDR3) is set forth in SEQ ID NO: 16; preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and / or a light chain variable region, wherein the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 18; and / or the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 17; preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is set forth in SEQ ID NO: 1, and / or the amino acid sequence of the light chain is set forth in SEQ ID NO: 2; Preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the heavy chain comprises a heavy chain variable region comprising 3 heavy chain complementarity determining regions (HCDRs), wherein the amino acid sequence of HCDR1 is set forth in SEQ ID NO: 19, the amino acid sequence of HCDR2 is set forth in SEQ ID NO: 20, and the amino acid sequence of HCDR3 is set forth in SEQ ID NO: 21; and / or the light chain comprises a light chain variable region comprising 3 light chain complementarity determining regions (LCDRs), wherein the amino acid sequence of LCDR1 is set forth in SEQ ID NO: 22, the amino acid sequence of LCDR2 is set forth in SEQ ID NO: 23, and the amino acid sequence of LCDR3 is set forth in SEQ ID NO: 24; preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and / or a light chain variable region, wherein the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 26; and / or the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 25; preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is set forth in SEQ ID NO: 3, and / or the amino acid sequence of the light chain is set forth in SEQ ID NO: 4; Preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the heavy chain comprises a heavy chain variable region comprising 3 heavy chain complementarity determining regions (HCDRs), wherein the amino acid sequence of HCDR1 is set forth in SEQ ID NO: 19, the amino acid sequence of HCDR2 is set forth in SEQ ID NO: 20, and the amino acid sequence of HCDR3 is set forth in SEQ ID NO: 21; and / or the light chain comprises a light chain variable region comprising 3 light chain complementarity determining regions (LCDRs), wherein the amino acid sequence of LCDR1 is set forth in SEQ ID NO: 22, the amino acid sequence of LCDR2 is set forth in SEQ ID NO: 23, and the amino acid sequence of LCDR3 is set forth in SEQ ID NO: 24; preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and / or a light chain variable region, wherein the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 26; and / or the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 25; preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is set forth in SEQ ID NO: 3, and / or the amino acid sequence of the light chain is set forth in SEQ ID NO: 4; Preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the heavy chain comprises a heavy chain variable region comprising 3 heavy chain complementarity determining regions (HCDRs), wherein the amino acid sequence of HCDR1 is set forth in SEQ ID NO: 27, the amino acid sequence of HCDR2 is set forth in SEQ ID NO: 28, and the amino acid sequence of HCDR3 is set forth in SEQ ID NO: 29; and / or the light chain comprises a light chain variable region comprising 3 light chain complementarity determining regions (LCDRs), wherein the amino acid sequence of LCDR1 is set forth in SEQ ID NO: 30, the amino acid sequence of LCDR2 is set forth in SEQ ID NO: 31, and the amino acid sequence of LCDR3 is set forth in SEQ ID NO: 32; preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and / or a light chain variable region, wherein the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 34; and / or the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 33; preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is set forth in SEQ ID NO: 5, and / or the amino acid sequence of the light chain is set forth in SEQ ID NO: 6; Preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the heavy chain comprises a heavy chain variable region comprising 3 heavy chain complementarity determining regions (HCDRs), wherein the amino acid sequence of HCDR1 is set forth in SEQ ID NO: 27, the amino acid sequence of HCDR2 is set forth in SEQ ID NO: 28, and the amino acid sequence of HCDR3 is set forth in SEQ ID NO: 29; and / or the light chain comprises a light chain variable region comprising 3 light chain complementarity determining regions (LCDRs), wherein the amino acid sequence of LCDR1 is set forth in SEQ ID NO: 30, the amino acid sequence of LCDR2 is set forth in SEQ ID NO: 31, and the amino acid sequence of LCDR3 is set forth in SEQ ID NO: 32; preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and / or a light chain variable region, wherein the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 34; and / or the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 33; preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is set forth in SEQ ID NO: 5, and / or the amino acid sequence of the light chain is set forth in SEQ ID NO: 6; Preferably, the antibody or antigen-binding fragment thereof is an anti-B7-H3 antibody or antigen-binding fragment thereof; preferably, the anti-B7-H3 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the heavy chain comprises a heavy chain variable region comprising 3 heavy chain complementarity determining regions (HCDRs), wherein the amino acid sequence of HCDR1 is set forth in SEQ ID NO: 35, the amino acid sequence of HCDR2 is set forth in SEQ ID NO: 36, and the amino acid sequence of HCDR3 is set forth in SEQ ID NO: 37; and / or the light chain comprises a light chain variable region comprising 3 light chain complementarity determining regions (LCDRs), wherein the amino acid sequence of LCDR1 is set forth in SEQ ID NO: 38, the amino acid sequence of LCDR2 is set forth in SEQ ID NO: 39, and the amino acid sequence of LCDR3 is set forth in SEQ ID NO: 40; preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and / or a light chain variable region, wherein the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 42; and / or the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 41; preferably, the anti-B7-H3 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is set forth in SEQ ID NO: 7, and / or the amino acid sequence of the light chain is set forth in SEQ ID NO: 8; Preferably, the antibody or antigen-binding fragment thereof is an anti-B7-H3 antibody or antigen-binding fragment thereof; preferably, the anti-B7-H3 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the heavy chain comprises a heavy chain variable region comprising 3 heavy chain complementarity determining regions (HCDRs), wherein the amino acid sequence of HCDR1 is set forth in SEQ ID NO: 35, the amino acid sequence of HCDR2 is set forth in SEQ ID NO: 36, and the amino acid sequence of HCDR3 is set forth in SEQ ID NO: 37; and / or the light chain comprises a light chain variable region comprising 3 light chain complementarity determining regions (LCDRs), wherein the amino acid sequence of LCDR1 is set forth in SEQ ID NO: 38, the amino acid sequence of LCDR2 is set forth in SEQ ID NO: 39, and the amino acid sequence of LCDR3 is set forth in SEQ ID NO: 40; preferably, the anti-NECTIN4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and / or a light chain variable region, wherein the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 42; and / or the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 41; preferably, the anti-B7-H3 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is set forth in SEQ ID NO: 7, and / or the amino acid sequence of the light chain is set forth in SEQ ID NO: 8; Preferably, the anti-B7-H3 antibody or antigen binding fragment thereof comprises a heavy chain and / or a light chain, wherein the heavy chain comprises a heavy chain variable region comprising 3 heavy chain complementarity determining regions (HCDRs), wherein the amino acid sequence of HCDR1 is set forth in SEQ ID NO: 43, the amino acid sequence of HCDR2 is set forth in SEQ ID NO: 44, and the amino acid sequence of HCDR3 is set forth in SEQ ID NO: 45; and / or the light chain comprises a light chain variable region comprising 3 light chain complementarity determining regions (LCDRs), wherein the amino acid sequence of LCDR1 is set forth in SEQ ID NO: 46, the amino acid sequence of LCDR2 is set forth in SEQ ID NO: 47, and the amino acid sequence of LCDR3 is set forth in SEQ ID NO: 48; preferably, the anti-NECTIN4 antibody or antigen binding fragment thereof comprises a heavy chain variable region and / or a light chain variable region, wherein the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 50; and / or the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 49; preferably, the anti-B7-H3 antibody or antigen binding fragment thereof comprises a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is set forth in SEQ ID NO: 9, and / or the amino acid sequence of the light chain is set forth in SEQ ID NO:

10. An antibody-drug conjugate, wherein, The compound of any one of claims 1-6 as a small molecule toxin moiety in a drug. An antibody-drug conjugate comprising a covalently attached antibody or antigen binding fragment thereof and a linker-small molecule toxin moiety, wherein, The drug linker conjugate of any one of claims 7-16 as a linker-small molecule toxin moiety in a drug. A pharmaceutical composition consisting of a substance X and a pharmaceutically acceptable excipient; The substance X is a substance X1 or a substance X2; the substance X1 is a compound of any one of claims 1-6 or a pharmaceutically acceptable salt thereof; the substance X2 is an antibody-drug conjugate of formula III according to any one of claims 17-22. The compound of any one of claims 1-6, the drug linker conjugate of any one of claims 7-16, the antibody-drug conjugate of any one of claims 17-22, or the pharmaceutical composition of claim 23 for use in the treatment or prevention of a disease associated with abnormal cell activity; preferably, the disease is a cancer; Preferably, wherein the cancer is a cancer associated with abnormal expression of B7-H3, HER2, NECTIN4, CDH17, or GPC3; Preferably, wherein the cancer is a cancer associated with abnormal expression of B7-H3, HER2, NECTIN4, CDH17, or GPC3; Preferably, the cancer is selected from breast cancer, gastric cancer, pancreatic cancer, prostate cancer, colorectal cancer, melanoma and lung cancer; preferably, the prostate cancer comprises androgen-dependent or androgen-independent prostate cancer; the lung cancer comprises non-small cell lung cancer and small cell lung cancer; the colorectal cancer comprises colorectal adenocarcinoma; the pancreatic cancer comprises pancreatic ductal adenocarcinoma. A method of preventing or treating a disease associated with abnormal cell activity, comprising administering to a subject in need thereof an effective amount of a compound of any one of claims 1-6, a drug linker conjugate of any one of claims 7-16, an antibody-drug conjugate of any one of claims 17-22, or a pharmaceutical composition of claim 23; preferably, the disease is cancer; Preferably, the cancer is associated with abnormal expression of B7-H3, HER2, NECTIN4, CDH17 or GPC3. Preferably, the cancer is selected from breast cancer, gastric cancer, pancreatic cancer, prostate cancer, colorectal cancer, melanoma and lung cancer; preferably, the prostate cancer comprises androgen-dependent or androgen-independent prostate cancer; the lung cancer comprises non-small cell lung cancer and small cell lung cancer; the colorectal cancer comprises colorectal adenocarcinoma; the pancreatic cancer comprises pancreatic ductal adenocarcinoma. A compound of any one of claims 1-6, a drug linker conjugate of any one of claims 7-16, an antibody-drug conjugate of any one of claims 17-22, or a pharmaceutical composition of claim 23 for use in preventing or treating a disease associated with abnormal cell activity; preferably, the disease is cancer. Preferably, the cancer is associated with abnormal expression of B7-H3, HER2, NECTIN4, CDH17 or GPC3; preferably, the cancer is selected from breast cancer, gastric cancer, pancreatic cancer, prostate cancer, colorectal cancer, melanoma and lung cancer; preferably, the prostate cancer comprises androgen-dependent or androgen-independent prostate cancer; the lung cancer comprises non-small cell lung cancer and small cell lung cancer; the colorectal cancer comprises colorectal adenocarcinoma; the pancreatic cancer comprises pancreatic ductal adenocarcinoma. Use of a compound of any one of claims 1-6 as a small molecule toxin for the preparation of an antibody drug conjugate. Use of a compound as described in any one of claims 7-15 for the manufacture of an antibody-drug conjugate comprising a covalently attached antibody or antigen binding fragment thereof and a linker-small molecule toxin moiety, wherein, The linker-small molecule toxin moiety is a drug linker conjugate of any one of claims 7-16.

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