Targeted protein degradation agent, and pharmaceutical composition and use thereof

Abstract

Disclosed in the present invention are a targeted protein degradation agent, and a pharmaceutical composition and the use thereof. The novel targeted protein degradation agent prepared in the present invention has significant target protein degradation activity. The novel targeted protein degradation agent of the present invention is used as a payload of an antibody-drug conjugate. By using the targeting property of the antibody and the antigen-mediated internalization effect, more effective and safer degradation of a target protein is achieved. The present invention can be used for treating target protein-mediated diseases (such as tumors), showing broad application prospects and research and development value in the pharmaceutical field.

Description

A targeted protein degrader, its pharmaceutical composition and application Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a targeted protein degrader, its pharmaceutical composition, and its application. Background Technology

[0002] Targeted protein degradation (TPD) is a novel and groundbreaking drug development strategy that utilizes inherent intracellular protein degradation pathways to directly degrade pathogenic target proteins. This novel drug formulation includes various types, such as PROTACs, molecular glues, LYTACs, ATACs, AbTACs, ATTECs, AUTACs, and AUTOTACs. Proteolysis-targeting chimeras (PROTACs) are heterobifunctional small molecule compounds composed of three parts: a ligand that binds to the target protein, a ligand that recruits E3 ligases, and a linker that helps anchor the target protein to the E3 ubiquitin ligase, promoting ubiquitination and subsequent proteasome degradation. Molecular glues (MGs) are small molecule compounds that differ from PROTACs in that they bind to E3 ligases and alter the shape of the ligase surface, enabling the ligase to bind to the target protein, thereby leading to the ubiquitination and degradation of the target.

[0003] Antibody-drug conjugates (ADCs) are composed of monoclonal antibodies targeting tumor-specific antigens or tumor-associated antigens, linked to varying numbers of small-molecule cytotoxic agents (or payloads) via linkers. They combine the high targeting specificity of monoclonal antibodies with the high activity of cytotoxic agents in tumor tissues, making them one of the fastest-growing drug classes in the field of targeted cancer therapy in recent years. Research has found that using protein degraders as the payload in ADCs can overcome the inherent limitations of these agents. Therefore, protein degraders are of great significance for drug development. Summary of the Invention

[0004] To overcome the shortcomings of the prior art, this invention discloses a targeted protein degrader, its pharmaceutical composition, and its applications. The inventors of this invention have creatively developed a targeted protein degrader, which, when conjugated with an antibody, forms a novel ADC drug capable of achieving cell-specific target protein degradation.

[0005] In a first aspect, the present invention provides a compound or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, or deuterated thereof having the following structure:

[0006] in,

[0007] U is selected from: C0-C 10 Alkyl, C3-C 10 cycloalkyl, C6-C 10 Aryl, 4-10 membered heterocyclic groups;

[0008] V is selected from: -O-, -S-,

[0009] B is

[0010] The C ring is a divalent group attached to the A ring; it either does not exist or is selected from: C3-C 10 Cycloalkylene, C6-C 10 arylene, 4-10 membered heterocyclic arylene;

[0011] X, Y, and Z are independently selected from: single bonds, C1-C 10 Alkylene, -(C0-C6 alkylene)-O-, -(C0-C6 alkylene)-S-, -(C0-C6 alkylene)-C(O)-, -(C0-C6 alkylene)-C(S)-, -(C0-C6 alkylene)-N(R) B1 )-、-(C0-C6 alkylene)-CON(R B1 )-、-(C0-C6 alkylene)-N(R B1 CO-, -(C0-C6 alkylene)-SO2-, -(C0-C6 alkylene)-SO-, -(C0-C6 alkylene)-(C3-C 10 Cycloalkylene)-, -(C0-C6 alkylene)-(C6-C 10 arylene)-, -(C0-C6 alkylene)-(4-10 heterocyclic)-; wherein, the C0-C 10 Alkylene, C3-C 10 Cycloalkylene, C6-C 10 The hydrogen atoms in the arylene and 4-10 membered heterocyclic groups may optionally be substituted with one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, -(C0-C6 alkylene)-(C3-C 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, C1-C 10Halogenated alkoxy groups, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)CON(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -O(C) 0-10 alkyl), -S(C 0-10 Alkyl), -SO(C) 0-10 alkyl), -SO2(C 0-10 alkyl), -SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0- 10 alkyl);

[0012] Ring A is selected from: C3-C 10 Cycloalkylene, C6-C 10 arylene, 4-10 membered heterocyclic; of which, C3-C 10 Cycloalkylene, C6-C 10 The hydrogen atoms in the arylene and 4-10 membered heterocyclic groups may optionally be substituted with one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, -(C0-C6 alkylene)-(C3-C 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, C1-C 10 Haloalkoxy, -(C0-C6 alkylene)-N(C 0-10 Alkyl)(C 0-10 Alkyl), -(C0-C6 alkylene)-N(C 0-10 Alkyl)CO(C 0-10 Alkyl), -(C0-C6 alkylene)-N(C 0-10 Alkyl)CON(C 0-10 Alkyl), -(C0-C6 alkylene)-N(C 0-10 Alkyl)SO2(C 0-10Alkyl), -(C0-C6 alkylene)-O(C 0-10 Alkyl), -(C0-C6 alkylene)-S(C 0-10 Alkyl), -(C0-C6 alkylene)-SO(C 0-10 Alkyl), -(C0-C6 alkylene)-SO2(C 0-10 Alkyl), -(C0-C6 alkylene)-SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -(C0-C6 alkylene)-COO(C 0-10 Alkyl), -(C0-C6 alkylene)-OCO(C 0-10 Alkyl), -(C0-C6 alkylene)-CON(C 0-10 Alkyl)(C 0-10 Alkyl), -(C0-C6 alkylene)-CO(C 0-10 alkyl);

[0013] D is

[0014] L1, L2, and L3 are independently selected from: single bonds, C1-C 10 Alkylene, -O-(C0-C6 alkylene)-, -S-(C0-C6 alkylene)-, -N(R L0 )-(C0-C6 alkylene)-, -N(R L0 )C(O)-(C0-C6 alkylene)-、-N(R L0 )C(O)O-(C0-C6 alkylene)-、-N(R L0 )C(O)N(R L0 -(C0-C6 alkylene)-, -C(O)-(C0-C6 alkylene)-, -C(S)-(C0-C6 alkylene)-, -C(O)O-(C0-C6 alkylene)-, -OC(O)-(C0-C6 alkylene)-, -(C0-C6 alkylene)-OC(O)-, -CON(R L0 -(C0-C6 alkylene)-, -SO2-(C0-C6 alkylene)-, -SO-(C0-C6 alkylene)-, C3-C 10 Cycloalkylene, C6-C 10 arylene, 4-10 membered heterocyclic alkylene; wherein, the CO-C 10 Alkylene, C3-C 10 Cycloalkylene, C6-C 10 The hydrogen atoms in the arylene and 4-10 membered heterocyclic groups may optionally be substituted with one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, C2-C 10alkenyl, C2-C 10 alkynyl, -(C0-C6 alkylene)-(C3-C 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, C1-C 10 Halogenated alkoxy groups, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)CON(C 0-10 alkyl), -N(C) 0- 10 Alkyl)SO2(C 0-10 Alkyl), -O(C) 0-10 alkyl), -S(C 0-10 Alkyl), -SO(C) 0-10 alkyl), -SO2(C 0-10 alkyl), -SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl);

[0015] L represents the connector precursor;

[0016] R V1 R V2 R B1 R L0 Independently selected from: H, deuterium, halogen, cyano, nitro, azide, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, -(C0-C6 alkylene)-(C3-C 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, C1-C 10 Halogenated alkoxy groups, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C)0-10 Alkyl)CON(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -O(C) 0-10 alkyl), -S(C 0-10 Alkyl), -SO(C) 0-10 alkyl), -SO2(C 0-10 alkyl), -SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl).

[0017] Specifically, the compound shown in Formula I contains at least one deuterium atom.

[0018] Specifically, U is selected from: C0-C6 alkyl, C3-C6 cycloalkyl, C6-C 10 Aryl, for example, H,

[0019] Specifically, U is a 5-6 member heterocyclic group, for example... Preferred

[0020] Specifically, W is selected from: -O-, -S-, Preferred

[0021] Specifically, R W1 R W2 Independently selected from: H, deuterium, halogen, cyano, nitro, azide, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, -(C0-C6 alkylene)-(C3-C 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, C1-C 10 Halogenated alkoxy groups, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)CON(C0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -O(C) 0-10 alkyl), -S(C 0-10 Alkyl), -SO(C) 0-10 alkyl), -SO2(C 0-10 alkyl), -SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl).

[0022] Specifically, R W1 R W2 Independently selected from: H, deuterium, halogen, cyano, nitro, azide, C1-C 10 Alkyl, C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl).

[0023] In some embodiments of the present invention, R W1 For H.

[0024] Specifically, W is selected from: -O-, -CH2-,

[0025] In some embodiments of the present invention, W is...

[0026] Specifically, U is selected from:

[0027] In some embodiments of the present invention, U is

[0028] Specifically, V is

[0029] Specifically, RV1 R V2 Independently selected from: H, deuterium, halogen, cyano, nitro, azide, C1-C 10 Alkyl, C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl).

[0030] In some embodiments of the present invention, R V1 For H.

[0031] In some embodiments of the present invention, R V2 For H.

[0032] In some embodiments of the present invention, V is -CH2- or...

[0033] Specifically, ring C either does not exist or is selected from:

[0034] Specifically, ring C is selected from:

[0035] In some embodiments of the present invention, the C-ring is absent or is [missing information].

[0036] Specifically, X is selected from: C1-C 10 Alkylene, -(C0-C6 alkylene)-O-, -(C0-C6 alkylene)-S-, -(C0-C6 alkylene)-C(O)-, -(C0-C6 alkylene)-C(S)-, -(C0-C6 alkylene)-N(R) X1 )-、-(C0-C6 alkylene)-CON(R X1 )-、-(C0-C6 alkylene)-N(R X1 )CO-, -(C0-C6 alkylene)-SO2-, -(C0-C6 alkylene)-SO-; wherein, R X1 Having the above R B1 Same definition.

[0037] Specifically, X is selected from: C1-C6 alkylene, -(C0-C6 alkylene)-N(R X1 )-.

[0038] Specifically, R X1 Selected from: H, deuterium, halogen, cyano, nitro, azide, C1-C 10 Alkyl, C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl).

[0039] In some embodiments of the present invention, R X1 For H.

[0040] Specifically, X is selected from:

[0041] In some embodiments of the present invention, X is selected from:

[0042] Specifically, Y is selected from: -(C0-C6 alkylene)-O-, -(C0-C6 alkylene)-S-, -(C0-C6 alkylene)-C(O)-, -(C0-C6 alkylene)-C(S)-, -(C0-C6 alkylene)-N(R) Y1 )-、-(C0-C6 alkylene)-CON(R Y1 )-、-(C0-C6 alkylene)-N(R Y1 CO-, -(C0-C6 alkylene)-SO2-, -(C0-C6 alkylene)-SO-, -(C0-C6 alkylene)-(C3-C 10 Cycloalkylene)-, -(C0-C6 alkylene)-(C6-C 10 arylene)-, -(C0-C6 alkylene)-(4-10 heterocyclic)-; wherein, the C0-C6 alkylene, C3-C 10 Cycloalkylene, C6-C 10 The hydrogen atoms in the arylene and 4-10 membered heterocyclic groups may optionally be substituted with one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, C1-C10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 Alkyl); R Y1 Having the above R B1 Same definition.

[0043] Specifically, Y is selected from: -(C0-C6 alkylene)-O-, -(C0-C6 alkylene)-N(R) Y1 )-、-(C0-C6 alkylene)-(C6-C 10 arylene)-, -(C0-C6 alkylene)-(4-10 heterocyclic)-; wherein, the C0-C6 alkylene, C6-C 10 The hydrogen atoms in the arylene and 4-10 membered heterocyclic groups may optionally be substituted with one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0-10 Alkyl), -O(C) 0-10 alkyl).

[0044] Specifically, R Y1 Selected from: H, deuterium, halogen, cyano, nitro, azide, C1-C 10 Alkyl, C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl).

[0045] In some embodiments of the present invention, R Y1 For H.

[0046] Specifically, Y is selected from: -O-,

[0047] In some embodiments of the present invention, Y is selected from: -O-,

[0048] Specifically, Z is selected from: -(C0-C6 alkylene)-O-, -(C0-C6 alkylene)-S-, -(C0-C6 alkylene)-C(O)-, -(C0-C6 alkylene)-C(S)-, -(C0-C6 alkylene)-N(R) Z1 )-、-(C0-C6 alkylene)-CON(R Z1 )-、-(C0-C6 alkylene)-N(R Z1 )CO-, -(C0-C6 alkylene)-SO2-, -(C0-C6 alkylene)-SO-; wherein, R Z1 Having the above R B1 Same definition.

[0049] Specifically, Z is selected from: -(C0-C6 alkylene)-O-, -(C0-C6 alkylene)-N(R) Z1 )-.

[0050] Specifically, R Z1 Selected from: H, deuterium, halogen, cyano, nitro, azide, C1-C 10 Alkyl, C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl).

[0051] In some embodiments of the present invention, R Z1 For H.

[0052] Specifically, Z is selected from: -O-,

[0053] In some embodiments of the present invention, Z is...

[0054] Specifically, B is selected from:

[0055] In some embodiments of the present invention, B is selected from:

[0056] Specifically, ring A is selected from:

[0057] Specifically, R L4 R L5 Independently selected from: deuterium, halogen, cyano, nitro, azide, C1-C 10 Alkyl, -(C0-C6 alkylene)-(C3-C6 alkylene) 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), -(C0-C6 alkylene)-N(C 0-10 Alkyl)(C 0-10 Alkyl), -(C0-C6 alkylene)-N(C 0-10 Alkyl)CO(C 0-10 Alkyl), -(C0-C6 alkylene)-O(C 0-10 Alkyl), -(C0-C6 alkylene)-COO(C 0-10 Alkyl), -(C0-C6 alkylene)-OCO(C 0-10 Alkyl), -(C0-C6 alkylene)-CON(C 0-10 Alkyl)(C 0- 10 Alkyl), -(C0-C6 alkylene)-CO(C 0-10 alkyl).

[0058] Specifically, R L4 R L5 Independently selected from: H, deuterium, F, Cl, Br, I,

[0059] Specifically, R L4 R L5 Independently selected from: H, Cl,

[0060] In some embodiments of the present invention, R L4 It can be H or Cl.

[0061] In some embodiments of the present invention, R L5 for

[0062] Specifically, ring A is selected from:

[0063] In some embodiments of the present invention, ring A is selected from:

[0064] Specifically, one or more H atoms bonded to the carbon atom in D can be replaced by deuterium.

[0065] Specifically, one or more H atoms bonded to the carbon atoms in L1, L2, and L3 can be replaced by deuterium.

[0066] Specifically, L1 is a divalent group attached to ring A, selected from: single bond, -O-(C0-C6 alkylene)-, -S-(C0-C6 alkylene)-, -N(R L1 )-(C0-C6 alkylene)-, -N(R L1 )C(O)-(C0-C6 alkylene)-、-N(R L1 )C(O)O-(C0-C6 alkylene)-、-N(R L1 )C(O)N(R L1 -(C0-C6 alkylene)-, -C(O)-(C0-C6 alkylene)-, -C(O)O-(C0-C6 alkylene)-, -CON(R L1 )-(C0-C6 alkylene)-; wherein, R L1 Having the above R L0 Same definition.

[0067] Specifically, L1 is selected from: single bond, -O-(C0-C6 alkylene)-, -N(R L1 )-(C0-C6 alkylene)-, -N(R L1 )C(O)O-(C0-C6 alkylene)-、-CON(R L1 )-(C0-C6 alkylene)-.

[0068] Specifically, R L1 Selected from: H, deuterium, halogen, cyano, nitro, azide, C1-C 10 Alkyl, C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C0-10 alkyl), -CO(C) 0-10 alkyl).

[0069] In some embodiments of the present invention, R L1 Selected from: H,

[0070] Specifically, L1 is selected from: single bond, -O-,

[0071] In some embodiments of the present invention, L1 is selected from: single bond, -O-, ...

[0072] Specifically, L2 is selected from: single bond, C1-C 10 Alkylene, -O-(C0-C6 alkylene)-, -S-(C0-C6 alkylene)-, -N(R L2 )-(C0-C6 alkylene)-, -N(R L2 -C(O)-(C0-C6 alkylene)-, -C(O)-(C0-C6 alkylene)-, -CON(R L2 )-(C0-C6 alkylene)-; wherein, the C0-C 10 The hydrogen in the alkylene group may optionally be substituted by one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, -(C0-C6 alkylene)-(C3-C6 alkylene) 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 Alkyl), -O(C) 0- 10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 Alkyl); wherein, R L2 Having the above R L0 Same definition.

[0073] Specifically, L2 is selected from: single bond, C1-C 10 Alkylene, -N(R) L2)-(C0-C6 alkylene)-; wherein, the C0-C 10 The hydrogen in the alkylene group may optionally be substituted by one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, -(C0-C6 alkylene)-(C3-C6 alkylene) 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0-10 Alkyl), -O(C) 0-10 alkyl).

[0074] Specifically, R L2 Selected from: H, deuterium, halogen, cyano, nitro, azide, C1-C 10 Alkyl, -(C0-C6 alkylene)-(C3-C6 alkylene) 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl).

[0075] In some embodiments of the present invention, R L2 Selected from:

[0076] Specifically, L2 is selected from: single bond,

[0077] In some embodiments of the present invention, L2 is selected from: single bonds,

[0078] Specifically, L3 is a divalent group attached to L, which is selected from: single bonds, C3-C bonds, etc. 10 Cycloalkylene, C6-C 10arylene, 4-10 membered heterocyclic alkylene; wherein, the C3-C 10 Cycloalkylene, C6-C 10 The hydrogen atoms in the arylene and 4-10 membered heterocyclic groups may optionally be substituted with one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0- 10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl).

[0079] Specifically, L3 is selected from: single bond, -(C0-C6 alkylene)-OC(O)-, C3-C 10 Cycloalkylene, 4-10 membered heterocyclic alkylene; wherein, the C0-C6 alkylene, C0-C6 alkylene, C6-C 10 The hydrogen atoms in the arylene and 4-10 membered heterocyclic groups may optionally be substituted with one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0- 10 Alkyl), -O(C) 0-10 alkyl).

[0080] Specifically, L3 is selected from: single key,

[0081] In some embodiments of the present invention, L3 is selected from: single bonds,

[0082] Specifically, D is selected from: -NH-,

[0083] In some embodiments of the present invention, D is selected from: -NH-,

[0084] In some embodiments of the present invention, D is selected from: -NH-,

[0085] Specifically, L is

[0086] Specifically, L4 is a divalent group attached to L3, and it is selected from: single bonds,

[0087] Specifically, L5 is a divalent group selected from: single bonds, C1-C6 alkylene groups, -N(C0-C5) groups, and C6(C6) groups. 10 Alkyl)-, -CO-, -O-, Wherein, s is an integer from 1 to 10 (specifically, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10); r is an integer from 1 to 10 (specifically, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10); P1, P2, P3, and P4 are either absent or amino acid residues (natural or non-natural amino acid residues of L- or D-configuration), and at least one of P1, P2, P3, and P4 is an amino acid residue.

[0088] Specifically, P1, P2, P3, and P4 independently do not contain or are selected from the following amino acid residues of L- or D-configuration: glycine residue, alanine residue, valine residue, leucine residue, isoleucine residue, methionine residue, proline residue, tryptophan residue, serine residue, tyrosine residue, cysteine ​​residue, phenylalanine residue, asparagine residue, glutamine residue, threonine residue, aspartic acid residue, glutamic acid residue, lysine residue, arginine residue, histidine residue, citrulline residue, ornithine residue, cysteine ​​residue, selenocysteine, hydroxyproline, hydroxylysine, and theanine.

[0089] Specifically, L6 is selected from: Where q is an integer from 1 to 10 (specifically, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10); R L3 R L6 R L7 Independently selected from: H, C1-C 10 Alkyl, -(C0-C6 alkylene)-(C6-C 10 Aryl).

[0090] Specifically, L is the uncuttable joint precursor, which is selected from: Where q is an integer from 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10). Specifically, L is a cleavable adapter precursor, for example, a protease-cleavable adapter precursor selected from: Where q is an integer from 1 to 10 (specifically 1, 2, 3, 4, 5, 6, 7, 8, 9, 10); P1, P2, P3, and P4 are either absent or amino acid residues (natural or non-natural amino acid residues of L- or D-configuration), and at least one of P1, P2, P3, and P4 is an amino acid residue.

[0091] Specifically, P1, P2, P3, and P4 independently do not contain or are selected from the following amino acid residues of L- or D-configuration: glycine residue, alanine residue, valine residue, leucine residue, isoleucine residue, methionine residue, proline residue, tryptophan residue, serine residue, tyrosine residue, cysteine ​​residue, phenylalanine residue, asparagine residue, glutamine residue, threonine residue, aspartic acid residue, glutamic acid residue, lysine residue, arginine residue, histidine residue, citrulline residue, ornithine residue, cysteine ​​residue, selenocysteine, hydroxyproline, hydroxylysine, and theanine.

[0092] Specifically, P1, P2, P3, and P4 are either absent or selected from: glycine residues, L-alanine residues, D-alanine residues, L-valine residues, D-valine residues, L-phenylalanine residues, D-phenylalanine residues, L-citrulline residues, D-citrulline residues, L-asparagine residues, and D-asparagine residues.

[0093] Specifically, P1 is a valine residue, P2 and P3 are not present, and P4 is a citrulline residue.

[0094] Specifically, P1, P2, and P4 are all glycine residues, and P3 is a phenylalanine residue.

[0095] Specifically, P1 is a valine residue, P2 and P3 are not present, and P4 is an alanine residue.

[0096] Specifically, P1, P2, and P3 are not present, and P4 is a citrulline residue.

[0097] Specifically, P1 is absent, P2 and P3 are both alanine residues, and P4 is an asparagine residue.

[0098] Specifically, L is selected from:

[0099] Specifically, L is selected from:

[0100] Where q is an integer from 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9).

[0101] In some embodiments of the present invention, q is 2.

[0102] In some embodiments of the present invention, q is 3.

[0103] In some embodiments of the present invention, q is 5.

[0104] In some embodiments of the present invention, q is 6.

[0105] Specifically, L is For example,

[0106] Specifically, L is a linker precursor that can be coupled to a ligand via a thiol group, and it is selected from:

[0107] Specifically, L is a linker precursor that can be coupled to a ligand via an amino group, and it is selected from:

[0108] Specifically, L is a linker precursor that can be coupled to a ligand via click chemistry, and it is selected from:

[0109] In some embodiments of the present invention, L has the following structure:

[0110] In some embodiments of the present invention, L can be, for example, the “MLE-” part in patent document WO2022166762A1, the “L1” part in WO2023025248A1, the precursor structure of “L” in CN115867322A, etc.

[0111] Specifically, the compound has the following structure:

[0112] Specifically, the compound has the following structure:

[0113] Specifically, the compound has the following structure:

[0114] Specifically, the compound has the following structure:

[0115] In some embodiments of the present invention, the compound has the following structure:

[0116] A second aspect of the present invention provides a conjugate (antibody-drug conjugate) or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, or deuterated thereof, said conjugate having the following structure:

[0117] Wherein, U, V, B, A ring, and D have the definitions described in the first aspect of this invention;

[0118] n is an integer between 1 and 10 (specifically 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) or a decimal (i.e., the DAR value of antibody-drug conjugates; when n is a decimal, it refers to the average number of linker-drug molecules in each Bm conjugate).

[0119] L' is the connector;

[0120] Bm is the connecting part.

[0121] Specifically, L is

[0122] Specifically, L4 and L5 have the definitions described in the first aspect of this invention.

[0123] Specifically, L6' is selected from: Where q is an integer from 1 to 10 (specifically, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10); R L3 Selected from: H, C1-C 10 Alkyl, -(C0-C6 alkylene)-(C6-C 10 Aryl).

[0124] Specifically, L' is a non-cuttable joint, which is selected from: Where q is an integer from 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10). Specifically, L' is a cleavable adapter, for example, a protease-cleavable adapter selected from: Where q is an integer from 1 to 10 (specifically 1, 2, 3, 4, 5, 6, 7, 8, 9, 10); P1, P2, P3, and P4 are either absent or amino acid residues (natural or non-natural amino acid residues of L- or D-configuration), provided that at least one of P1, P2, P3, and P4 is an amino acid residue.

[0125] Specifically, P1, P2, P3, and P4 independently do not contain or are selected from the following amino acid residues of L- or D-configuration: glycine residue, alanine residue, valine residue, leucine residue, isoleucine residue, methionine residue, proline residue, tryptophan residue, serine residue, tyrosine residue, cysteine ​​residue, phenylalanine residue, asparagine residue, glutamine residue, threonine residue, aspartic acid residue, glutamic acid residue, lysine residue, arginine residue, histidine residue, citrulline residue, ornithine residue, cysteine ​​residue, selenocysteine, hydroxyproline, hydroxylysine, and theanine.

[0126] Specifically, P1, P2, P3, and P4 are either absent or selected from: glycine residues, L-alanine residues, D-alanine residues, L-valine residues, D-valine residues, L-phenylalanine residues, D-phenylalanine residues, L-citrulline residues, D-citrulline residues, L-asparagine residues, and D-asparagine residues.

[0127] Specifically, P1 is a valine residue, P2 and P3 are not present, and P4 is a citrulline residue.

[0128] Specifically, P1, P2, and P4 are all glycine residues, and P3 is a phenylalanine residue.

[0129] Specifically, P1 is a valine residue, P2 and P3 are not present, and P4 is an alanine residue.

[0130] Specifically, P1, P2, and P3 are not present, and P4 is a citrulline residue.

[0131] Specifically, P1 is absent, P2 and P3 are both alanine residues, and P4 is an asparagine residue.

[0132] Specifically, L' is selected from:

[0133] Specifically, L' is selected from:

[0134] q is an integer from 1 to 10 (specifically, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10).

[0135] In some embodiments of the present invention, q is 2.

[0136] In some embodiments of the present invention, q is 3.

[0137] In some embodiments of the present invention, q is 5.

[0138] In some embodiments of the present invention, q is 6.

[0139] Specifically, L' is For example,

[0140] Specifically, L' is a linker precursor that can be coupled to a ligand via a thiol group, and it is selected from:

[0141] Specifically, L' is a linker precursor that can be coupled to a ligand via an amino group, and it is selected from: Specifically, L' is a linker precursor that can be coupled to a ligand via click chemistry, and it is selected from:

[0142] In some embodiments of the present invention, L' has the following structure:

[0143] In some embodiments of the present invention, L can be, for example, the divalent group form of “MLE-” in patent document WO2022166762A1, the divalent group form of “L1” in WO2023025248A1, the “L” part in CN115867322A, etc.

[0144] Specifically, Bm is a binding portion capable of specifically binding to a protein, wherein the binding portion is an antibody, an antibody fragment, or an antigen-binding fragment.

[0145] Specifically, Bm represents the antibody or antigen-binding portion.

[0146] Specifically, the protein bound to the binding portion is a surface antigen.

[0147] Specifically, the surface antigen is selected from 5T4, ACE, ADRB3, AKAP-4, ALK, androgen receptor, AOC3, APP, axonin 1, AXL, B7H3, B7-H4, BCL2, BCMA, bcr-abl, BORIS, BST2, C242, C4.4a, and CA. 125, CA6, CA9, CAIX, CCL11, CCR5, CD123, CD133, CD138, CD142, CD15, CD15-3, CD171, CD179a, CD18, CD19, CD19-9, CD2, CD20, CD22, CD23, CD24, CD25, CD27L, CD28, CD3, CD30, CD31, CD300LF, CD33, CD352, CD37, CD38, CD4, CD40, CD41, CD44, CD44v6, CD5 CD51, CD52, CD54, CD56, CD62E, CD62P, CD62L, CD70, CD71, CD72, CD74, CD79a, CD79b, CD80, CD90, CD97, CD125, CD138, CD141, CD147, CD152, CD154, CD326, CEA, CEACAM5, CFTR, condensation factor, cKit, closure protein 3, closure protein 18.2, CLDN6, CLEC12A, CLL-1, cll3, c-MET, Crypto 1. Growth factors, CS1, CTLA-4, CXCR2, CXORF61, cyclin B1, CYP1B1, cadherin-3, cadherin-6, DLL3, E7, EDNRB, EFNA4, EGFR, EGFRvIII, ELF2M, EMR2, ENPP3, EPCAM, EphA2, liver glycoside A4, liver glycoside B2, EPHB4, ERBB2 (Her2 / neu), ErbB3, ERG (TMPRSS2ETS fusion gene), E TBR, ETV6-AML, FAP, FCAR, FCRL5, FGFR1, FGFR2, FGFR3, FGFR4, FLT3, folate receptor α, folate receptor β, FOLR1, Fos-associated antigen 1, fucose GM1, GCC, GD2, GD3, GloboH, GM3, GPC1, GPC2, GPC3, gp1OO, GPNMB, GPR20, GPRC5D, GUCY2C, HAVCR1, HER2, HER3, HGF, HMI.24. HMWMAA, HPV E6, hTERT, human telomerase reverse transcriptase, ICAM, ICOS-L, IFN-α, IFNγ, IGF-I receptor, IGLL1, IL-2 receptor, IL-4 receptor, IL-13Ra2, IL-11Ra, IL-1, IL-12, IL-23, IL13, IL-22, IL-4, IL-5, IL-6, interferon receptor, integrins (including α4, α). v β3, α v β5, α v β6, α1β4, α4β1, α4β7, α5β1, α6β4, α IIbβ3 integrin), integrin αV, intestinal carboxyl esterase, KIT, LAGE-1a, LAIR1, LAMP-1, LCK, Legumain, LewisY, LFA-1 (CD11a), L-selectin (CD62L), LILRA2, LIV-1, LMP2, LRRC15, LY6E, LY6K, LY75, MAD-CT-1, MAD-CT-2, MAGE A1, MelanA / MART1, mesothelin, ML-IAP, MSLN, mucin, MUC1, MUC16, mut hsp70-2, MYCN, myostatin, NA17, NaPi2b, NCA-90, NCAM, Nectin-4, NGF, NOTCH1, NOTCH2, NOTCH3, NOTCH4, NY-BR-1, NY-ESO-1, o-acetyl-GD2, OR51E2, OY-TES1, p53, p53 mutant, PANX3, PAP, PAX3, PAX5, p-CAD, PCTA-1 / galactagoguerin 8, PD-L1, PD-L2, PDGFR, PDGFR-β, phosphatidylserine, PIK3CA, PLAC1, polysialic acid, prostate enzymes, prostate cancer cells, prostate Adenosine, Pseudomonas aeruginosa, rabies virus, survivin and telomerase, PRSS21, PSCA, PSMA, PTK7, RAGE-1, RANKL, Ras mutant, respiratory syncytial virus, rhesus monkey factor, RhoC, RON, ROR1, ROR2, RU1, RU2, sarcoma translocation breakpoint, SART3, SLAMF7, SLC44A4, sLe, SLITRK6, spermin 17, sphingosine 1-phosphate, SSEA-4, SSX2, STEAP1, TAG72, TARP, TCRβ, TEM1 / CD248, TEM7R, tendinin C, TF, TGF-1, TGF-β2, TNF-α, TGS5, Tie 2. One or more of the following: TIM-1, TnAg, TRAC, TRAIL-R1, TRAIL-R2, TROP-2, TRP-2, TRPV1, TSHR, tumor antigen CTAA16.88, tyrosinase, UPK2, VEGF, VEGFR1, VEGFR2, vimentin, WT1, XAGE1, B7H3, LAG-3 (CD223), PD-1 / PD-L1, BTLA, TIM-3, AA2R, CEACAM1, SIRPα, CD200R, and c-met.

[0148] Specifically, the surface antigen is selected from one or more of HER2, B7H3, CD20, CD38, CD33, BCMA, CD138, EGFR, FGFR4, GD2, PDGFR, TEM1 / CD248, TROP-2, PD-L1, CD123, and c-met.

[0149] In some embodiments of the present invention, the surface antigen is HER2 or B7H3.

[0150] Specifically, the antibodies are selected from: 3F8, 8H9, abagovomab, abciximab, abituzumab, abrezekimab, abrilumab, actoxumab, adalimumab, adecatumumab, aducanumab, aafasevikumab, afelimomab, afutuzumab, and alacximab. (izumab), ALD518, alemtuzumab, alirocumab, attumomab, amatuximab, anatumomab, andecaliximab, anetumab, anifrolumab, anrukinzumab, apolizumab, aprutumab, arcitumomab, ascrinvacumab Aselizumab, Atidortoxumab, Atlizumab, Tocilizumab, Atezolizumab, Atinumab, Atorolimab, Avelumab (Bavencio), Azintuxizumab, Belantan, Bapineuzumab, Bavituximab, B CD-100, bectumomab, begelomab, belantamab, belimumab, bemarituzumab, benalizumab, bermekimab, bersanlimab, bertilimumab, besilesomab, bevacizumab, bezlotoxumab, biciromabBimagrumab, bimekizumab, birtamimab, bivatuzumab, bleselumab, blinatumomab, blontuvetmab, blosozumab, bococizumab, brazikumab, brentuximab, briakinumab, brodalumab lumab (SILIQTM), brolucizumab, brontictuzumab, burosumab, cabiralizumab, caplacizumab, camidanlumab, camrelizumab, canakinumab, cantuzumab, capromab, carlumab, carotuximab b) Catumaxomab, cBR96, CC49, cedelizumab, cemiplimab, cergutuzumab, certrelimab, certolizumab, cetuximab, cibisatamab, cirmtuzumab, citatuzumab, cixutumumab, clazakizumab Clinoliximab, clivatuzumab, codrituzumab, cofetuzumab, coltuximab, conatumumab, concizumab, cosfroviximab, CR6261, crenezumab, crizanlizumab, crotedumab, cusatuzumabDacetuzumab, daclizumab, dalotuzumab, dapirolizumab, daratumumab, dectrekumab, demcizumab, denintuzumab, denosumab, depatuxizumab, derlotuximab, detumomab, dezamizumab mizumab, dinutuximab, diridavumab, domagrozumab, dostarlimab, dorlimomab, dolixizumab, drozitumab, duligotuzumab, dupilumab, durvalumab, dusigitumab, ecoromeximab, ikulimab eculizumab, edobacomab, edrecolomab, efalizumab, efungumab, eldelumab, elezanumab, elgemtumab, elotuzumab, elsilimomab, emactuzumab, emapalumab, emibetuzumab, emicizumab Entamab, enapotamab, enavatuzumab, enfortumab, enlimomab, enoblituzumab, enokizumab, enoticumab, ensituximab, epitumomab, eptinezumab, epratuzumab, erenumab, erlizumabErtumaxomab, etaracizumab, etigilimab, etrolizumab, evinacumab, evolocumab, exbivirumab, fanolesomab, faralimomab, faricimab, farletuzumab, farsinumab, FBTA05, felvizuma b) Fezakinumab, Fibatuzumab, Ficlatuzumab, Figitumumab, Frivumab, Flanvotumab, Fletikumab, Flotetuzumab, Fontolizumab, Foralumab, Foravirumab, Fremanezumab, Fres olimumab, frovocimab, frunevetmab, fulranumab, futuximab, galcanezumab, galiximab, gancotamab, ganitumab, gantenerumab, gavilimomab, gedivumab, gemtuzumab, geokizumab Gilvetmab, Gimsilumab, Girentuximab, Glembatumumab, Golimumab, Gomiliximab, Gusselkumab, HuMy9-6, OR000213, Ianalumab, Ibalizumab, IBI308, Ibrimomab, Icrucumab, IdarucizumabIfabotuzumab, igovomab (INDIMACIS-125), iladatuzumab, IMAB362, imalumab, imaprelimab, imciromab, imgatuzumab, inclacumab, indatuximab, indusatumab, inebilizumab, and infliximab are among the drugs used. Intetumumab, Inolitumumab, Inotuzumab, Iomab-B, Ipilimumab, Iratumumab, Isataximab, Icalimab, Istiratumab, Itolizumab, Ixekizumab, Keliximab, Labetuzumab (CEA-CIDETM), Latozumab (l Acnotuzumab, Ladiratuzumab, Lampalizumab, Lanadelumab, Landogrozumab, Lapriximab, Larcaviximab, Lebrikizumab, Lemeresomab, Lendalizumab, Lenvervimab, Lenzilumab, Ledalizumab (The following are listed as examples of specific abbreviations:) erdelimumab, leronlimab, lesofavumab, letolizumab, lexatumumab, libivirumab, lidostuzumab, ligelizumab, lilotomab, lintozumab, lirilumab, lodelcizumab, lokivetmab, and loncastuximab.Lovotuzumab, Losatuxizumab, Lucatumumab, Lulizumab, Lumiliximab, Lumretuzumab, Lupartumab, Lutikizumab, Mapatumumab, Margetuximab, Marstacimab, Maslimomab, Matuzumab (The following are listed as examples of specific abbreviations): umab, mavrilimumab, mepolizumab, metelimumab, milatuzumab, minretumomab, mirikizumab, mirvetuximab, mitumomab, modotuximab, mogalizumab, mogamulizumab, morolimumab. Mosunetuzumab, motavizumab, moxetumomab, muromonab-CD3 (ORTHOCLONE), napromab, namilumab, naptumomab, naratuximab, namatumab, natalizumab, navicixizumab, navivumab, nacituzumab (naxitamab), nebacumab, necitumumab, nemolizumab, NEOD001, nerelimomab, nesvacumab, netakimab, nimotuzumab, nirsevimab, nivolumab, nofetumomab, obiltoxaximab, obinutuzumabOcaratuzumab, ocrelizumab, odulimomab, ofatumumab, olaratumab, oleclumab, olendalizumab, olokizumab, omalizumab, omburtamab, OMS721, onartu zumab), ontecizumab, ontuxizumab, onvatilimab, opicinumab, oportuzumab, oregovomab (OVAREX), orticumab, otelixizumab, otilimab, otlertuzumab, Oktar Oxelumab, ozanezumab, ozoralizumab, pagibaximab, palivizumab, pamrevlumab, panitumumab, pankomab, panobacumab, parsatuzumab, and pascolizumab are listed as potential alternatives to oxelumab, ozanezumab, ozoralizumab, pagibaximab, and pascolizumab. The following are listed: pasotuxizumab, pateclizumab, patritumab, PDR001, pembrolizumab, pemtumomab, perakizumab, pertuzumab, pexelizumab, pidilizumab, pinatuzumab vedotin, pintumomab, placulumab, plozalizumab (Polivy), prezalumab, plozalizumab, pogalizumab, and ponezumab.Porgaviximab, prasinezumab, prezalizumab, priliximab, pritoxaximab, pritumumab, PRO 140. Quilizumab, Racotumomab, Radretumab, Rafivirumab, Ralpancizumab, Ramucirumab, Ranevetmab, Ranibizumab, Ravagalimab, Ravalizumab, Raxibacumab, Refanezumab, Reregavirumab, REGN-EB3, Renatlimab, Remtolumab, Reslizumab, Rilotumumab, Rinucumab, Risanki zumab, rituximab, rivabazumab, rmab, robatumumab, rodedumab, romilkimab, romosozumab, rontalizumab, rosmantuzumab, rovalpituzumab, rovelizumab, rozanolixizumab, ruplizumab (ANTOVA), SA237, sacituzumab, samalizumab, samrotamab, sarilumab, satralizumab, satumomab Pendetide, secukinumab, selicrelumab, seribantumab, setoxaximab, setrusumab, sevirumab, SGN-CD19A, SHP647Sibrotouzumab, sifalimumab, siltuximab, simtuzumab, siplizumab, sirtratumab, sirukumab, sofituzumab, solanezumab, solitomab, sonepcizumab, sontuzumab, spartalizumab, stamurorumab tamulumab), STI-6129, sulesomab, suptavumab, sutimlimab, suvizumab, suvratoxumab, tabalumab, tacatuzumab, tadocizumab, talacotuzumab, talizumab, tamtuvetmab, tanezumab, taplitumomab Paptox, tarextumab, tavolimab, tefibazumab, telimomab, telisotuzumab, tesidolumab, tetraxetan, tetulomab, tenatumomab, teneliximab, teprotumumab, teplizumab, etc. Tezepelumab, TGN1412, tibulizumab, ticilimumab, tigatuzumab, timigutuzumab, timolumab, tiragolumab, tiragotumab, tislelizumab, tisotumab, tiuxetan, tildrakizumabTNX-650, tomuzotuximab, toralizumab, tosatoxumab, tositumomab, tovetumab, tralokinumab, trastuzumab TRBS07, tregalizumab, tremelimumab, trevogrumab, tucotuzumab, tuvirumab, utosa Urtoxazumab, Ustekinumab, Ublituximab, Ulocuplumab, Urelumab, Utomilumab, Vadastuximab, Vanallimab, Vandortuzumab, Vantictumab, Vanucizumab, Vapaliximab, Varixa cumab), varlilumab, vatelizumab, vedolizumab, veltuzumab, vepalimomab, vesencumab, visilizumab, vobarilizumab, volociximab, vonlerolizumab, vopratelimab, vorse Tuzumab, votumumab, vunakizumab, xentuzumab, XMAB-5574, zalutumumab (HuMEX-EGFr), zanolimumab (HuMAX-CD4), zatuximab, zenocutuzumab, ziralimumab, zolbetuximab, zolimomab, or anti-B7-H3 antibody.

[0151] Specifically, the antibodies are selected from: rituximab, trastuzumab, gemtuzumab, pertuzumab, obinutuzumab, ofatumumab, olaratumab, ontuximab, isatuximab, sacituzumab, U3-1784, daratumumab, STI-6129, OR000213, lintuzumab, huMy9-6, balantamab, indatuximab, cetuximab, dinutuximab, and anti-CD38. A2 antibody, HuaAT13 / 5 antibody, alemtuzumab, ibritumomab, tositumomab, bevacizumab, panitumumab, tremelimumab, ticilimumab, catumaxomab, oregovomab, veltuzumab, anti-B7-H3 antibody.

[0152] Specifically, the antibody is rituximab, trastuzumab, pertuzumab, OR000213, lintuzumab, gemtuzumab, or anti-B7-H3 antibody.

[0153] In some embodiments of the present invention, the antibody is trastuzumab (such as Herceptin, whose heavy chain variable region and light chain variable region sequences are shown as SEQ ID NO:1 and SEQ ID NO:3, respectively, and whose heavy chain constant region and light chain constant region sequences are shown as SEQ ID NO:2 and SEQ ID NO:4, respectively).

[0154] In some embodiments of the present invention, the antibody is an anti-B7-H3 antibody (such as Ifinatamab, whose heavy chain variable region and light chain variable region sequences are shown as SEQ ID NO:5 and SEQ ID NO:7, respectively, and whose heavy chain constant region and light chain constant region sequences are shown as SEQ ID NO:6 and SEQ ID NO:8, respectively).

[0155] Specifically, the conjugate has the following structure:

[0156] Specifically, the compound has the following structure:

[0157] Specifically, the conjugate has the following structure:

[0158] Specifically, the conjugate has the following structure: (The linking sites on Bm are the thiol sites contained therein).

[0159] In some embodiments of the present invention, the conjugate has the following structure:

[0160] In some embodiments of the invention, the conjugate has the structure shown in L-1, wherein the Bm portion is Herceptin, and n = 5-6, such as 5.6 (as described in the example ADC24).

[0161] In some embodiments of the invention, the conjugate has the structure shown in L-4, wherein the Bm portion is Herceptin, n = 3-4, 5-6, 7-8, 8-9, such as 3.6, 3.7, 3.8, 4.0, 5.4, 7.3, 7.6, 7.8, 8.0 or 8.1 (as described in the examples ADC01, ADC02, ADC12, ADC19, ADC20, ADC22, ADC25 to ADC28).

[0162] In some embodiments of the present invention, the conjugate has the structure shown in L-4, wherein the Bm portion is Ifinatamab, n = 3-4, 6-7, such as 3.9 or 6.5 (as described in the examples ADC03, ADC04).

[0163] In some embodiments of the present invention, the conjugate has the structure shown in L-3, wherein the Bm portion is Herceptin, n = 3-4, 4-5, 7-8, such as 3.7, 3.8, 4.8 or 7.8 (as described in the examples ADC09, ADC09, ADC15, ADC29).

[0164] In some embodiments of the invention, the conjugate has the structure shown in L-5, wherein the Bm portion is Herceptin, n = 2-3, such as 2.0 (as described in the examples of ADC10).

[0165] In some embodiments of the invention, the conjugate has the structure shown in L-6, wherein the Bm portion is Herceptin, n = 1-3, such as 2.0 (ADC11 as described in the examples).

[0166] In some embodiments of the invention, the conjugate has the structure shown in L-52, wherein the Bm portion is Herceptin, n = 3-4, such as 3.05 (as described in the example ADC13).

[0167] In some embodiments of the invention, the conjugate has the structure shown in L-53, wherein the Bm portion is Herceptin, n = 3-4, such as 3.53 (as described in the example ADC16).

[0168] In some embodiments of the invention, the conjugate has the structure shown in L-54, wherein the Bm portion is Herceptin, n = 3-4, such as 3.50 (as described in the example ADC17).

[0169] In some embodiments of the invention, the conjugate has the structure shown in L-55, wherein the Bm portion is Herceptin, n = 2-3, such as 2.19 (ADC14 as described in the examples).

[0170] A third aspect of the present invention provides a method for preparing the compound described in the first aspect of the present invention, the method comprising the following steps: […]. With LR G 'Coupling; where R G R G ' is an optional, suitable reactive group.

[0171] In some embodiments of the present invention, R G For H.

[0172] In some embodiments of the present invention, R G 'for -OH,

[0173] A fourth aspect of the present invention provides a method for preparing the conjugate described in the second aspect of the present invention, the method comprising the following steps: [The method involves...] With Bm-R G "Coupled; where R" G " " represents any suitable reactive group.

[0174] A fifth aspect of the present invention provides a pharmaceutical composition comprising the compound described in the first aspect of the present invention or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate or deuterated thereof, or the conjugate described in the second aspect of the present invention or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate or deuterated thereof, and one or more pharmaceutically acceptable excipients.

[0175] Specifically, the pharmaceutically acceptable excipients may be selected from one or more of the following: fillers, binders, lubricants, disintegrants, suspending agents, solubilizers, thickeners, stabilizers, preservatives, antioxidants, buffers, and antibacterial agents.

[0176] Specifically, the pharmaceutical composition can be administered via any suitable route of administration, such as gastrointestinal administration (e.g., oral, sublingual, rectal administration) or non-gastrointestinal administration (e.g., intravenous, intramuscular, intranasal, intraocular, intracerebral, intravaginal, intraperitoneal, transdermal, subcutaneous, intradermal, respiratory tract administration, etc.).

[0177] Specifically, the pharmaceutical composition can be prepared into any suitable pharmaceutical dosage form: oral dosage forms (e.g., tablets, pills, powders, granules, capsules, lozenges, syrups, liquids, emulsions, suspensions, etc.); injections (e.g., for subcutaneous injection, intravenous injection, intramuscular injection, intraperitoneal injection), respiratory dosage forms (e.g., sprays, aerosols, powder sprays, etc.), skin dosage forms (e.g., topical solutions, lotions, ointments, plasters, pastes, patches, etc.), mucosal dosage forms (e.g., eye drops, ophthalmic ointments, nasal drops, mouthwashes, sublingual tablets, etc.), and cavity dosage forms (e.g., suppositories, aerosols, effervescent tablets, drops, pills, etc., for rectal, vaginal, urethral, ​​nasal, ear canal, etc.).

[0178] Specifically, the pharmaceutical composition is preferably in unit dosage form. In this form, the formulation is further divided into unit doses containing an appropriate amount of the active ingredient. The unit dosage form can be a capsule, tablet, or any dosage form; alternatively, the unit dosage form can also be a packaged formulation, such as tablets, capsules, and powders packaged in vials or ampoules.

[0179] Specifically, in the pharmaceutical composition, the compound (or conjugate) or its pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate or deuterated form may be used alone or in combination with other types of active ingredients.

[0180] Specifically, the amount of the active ingredient in the unit dose formulation may be varied or adjusted from 0.1 mg to 1000 mg (e.g., 0.1, 1, 5, 10, 20, 40, 50, 100, 200, 400, 500, 1000 mg), depending on the specific application and potency of the active ingredient. If desired, the composition may also contain other suitable therapeutic agents.

[0181] A sixth aspect of the invention provides the use of the compounds described in the first aspect of the invention or pharmaceutically acceptable salts, stereoisomers, esters, prodrugs, solvates, or deuterated derivatives thereof, or the conjugates described in the second aspect of the invention or pharmaceutically acceptable salts, stereoisomers, esters, prodrugs, solvates, or deuterated derivatives thereof, or the pharmaceutical compositions described in the third aspect of the invention, in the preparation of medicaments for the prevention and / or treatment of diseases related to target proteins.

[0182] Specifically, the disease is one that can be prevented and / or treated by inhibiting or degrading a target protein, such as the Bm-specifically bound protein described in the second aspect of the present invention. The disease includes, but is not limited to, tumors, cardiovascular and cerebrovascular diseases, and viral infection-related diseases.

[0183] Specifically, the tumors are selected from: liver cancer (such as hepatocellular carcinoma), lung cancer (such as non-small cell lung cancer, small cell lung cancer), gastric cancer, breast cancer (such as ductal carcinoma of the breast), colon cancer, bile duct cancer, bladder cancer, head and neck cancer (such as squamous cell carcinoma of the head and neck), cervical cancer, ovarian cancer, prostate cancer, thyroid cancer (such as papillary thyroid carcinoma), squamous cell carcinoma, lymphoma (such as non-Hodgkin lymphoma, Hodgkin lymphoma), sarcoma, acute myeloid leukemia, chronic lymphocytic leukemia, multiple myeloma, and myelodysplastic syndrome. More specifically, the tumors can be tumors related to abnormal expression of HER2 and B7H3.

[0184] In some embodiments of the present invention, the tumor is gastric cancer.

[0185] In some embodiments of the present invention, the tumor is breast cancer.

[0186] In some embodiments of the present invention, the tumor is ovarian cancer.

[0187] Specifically, the viral infection-related diseases are selected from: AIDS, hepatitis B, hepatitis C, hepatitis A, influenza, Japanese encephalitis, herpes, etc.

[0188] In a seventh aspect of the invention, a method for inhibiting or degrading a target protein is provided, comprising the steps of using a compound described in the first aspect of the invention or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, or deuterated thereof, or a conjugate described in the second aspect of the invention or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, or deuterated thereof, or a pharmaceutical composition described in the third aspect of the invention.

[0189] Specifically, the method is performed in vivo or in vitro.

[0190] In an eighth aspect of the invention, a method for preventing and / or treating target protein-related diseases is provided, comprising administering to a subject in need a compound of the first aspect of the invention or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, or deuterated thereof, or a conjugate of the second aspect of the invention or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, or deuterated thereof, or a pharmaceutical composition of the third aspect of the invention.

[0191] Specifically, the disease is as described in the sixth aspect of the present invention.

[0192] Specifically, the subjects were mammals, particularly humans.

[0193] Specifically, the administration can be carried out via any suitable route of administration, such as gastrointestinal administration (e.g., oral, sublingual, rectal administration) or non-gastrointestinal administration (e.g., intravenous, intramuscular, intranasal, intraocular, intracerebral, intravaginal, intraperitoneal, transdermal, subcutaneous, intradermal, respiratory tract administration, etc.).

[0194] In a ninth aspect of the invention, a method for treating tumors is provided, comprising administering to a subject in need a compound of the first aspect of the invention or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, or deuterated thereof, or a conjugate of the second aspect of the invention or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, or deuterated thereof, or a pharmaceutical composition of the third aspect of the invention.

[0195] Specifically, the tumor is as described in the sixth aspect of the present invention, for example, a tumor associated with abnormal expression of HER2 and B7H3.

[0196] The inventors of this invention have creatively discovered a novel targeted protein degrader that can be used as a payload for antibody-drug conjugates. By utilizing the targeting properties of antibodies and antigen-mediated internalization, it achieves more effective and safer degradation of target proteins for the treatment of target protein-mediated diseases (such as tumors), showing broad application prospects and research and development value in the pharmaceutical field. Attached Figure Description

[0197] Figure 1 shows the SDS-PAGE analysis results of Ifinatamab antibody.

[0198] Figure 2 shows the SEC analysis results of Ifinatamab antibody.

[0199] Figure 3 shows the SEC analysis results of the ADC01 (HER-LP03-Dar3.74) sample.

[0200] Figure 4 shows the SEC analysis results of the ADC02 (HER-LP03-Dar8.06) sample.

[0201] Figure 5 shows the SEC analysis results of the ADC03 (IFI-LP03-Dar3.92) sample.

[0202] Figure 6 shows the SEC analysis results of the ADC04 (IFI-LP03-Dar6.52) sample.

[0203] Figure 7 shows the SEC analysis results of the ADC05 (HER-LP07-Dar3.97) sample.

[0204] Figure 8 shows the SEC analysis results of the ADC06 (HER-LP07-Dar6.39) sample.

[0205] Figure 9 shows the SEC analysis results of the ADC07 (HER-LP07-Dar8.05) sample.

[0206] Figure 10 shows the SEC analysis results of the ADC08 (HER-LP04-Dar3.75) sample.

[0207] Figure 11 shows the SEC-MS analysis results of the ADC01 (HER-LP03-Dar3.74) sample.

[0208] Figure 12 shows the SEC-MS analysis results of the ADC02 (HER-LP03-Dar8.06) sample.

[0209] Figure 13 shows the SEC-MS analysis results of the ADC03 (IFI-LP03-Dar3.92) sample.

[0210] Figure 14 shows the SEC-MS analysis results of the ADC04 (IFI-LP03-Dar6.52) sample.

[0211] Figure 15 shows the SEC-MS analysis results of the ADC05 (HER-LP07-Dar3.97) sample.

[0212] Figure 16 shows the SEC-MS analysis results of the ADC06 (HER-LP07-Dar6.39) sample.

[0213] Figure 17 shows the SEC-MS analysis results of the ADC07 (HER-LP07-Dar8.05) sample.

[0214] Figure 18 shows the SEC-MS analysis results of the ADC10 (HER-LP05-Dar2.03) sample.

[0215] Figure 19 shows the SEC-MS analysis results of the ADC11 (HER-LP06-Dar1.97) sample.

[0216] Figure 20 shows the SEC-MS analysis results of the ADC08 (HER-LP04-Dar3.75) sample.

[0217] Figure 21 shows the SEC-MS analysis results of the ADC09 (HER-LP04-Dar7.83) sample.

[0218] Figure 22 shows the HIC analysis results of the ADC01 (HER-LP03-DAR3.74) sample.

[0219] Figure 23 shows the HIC analysis results of the ADC08 (HER-LP04-Dar3.75) sample.

[0220] Figure 24 shows the experimental results of the antiproliferative activity of small molecule compounds in N87 cells.

[0221] Figure 25 shows the experimental results of the antiproliferative activity of small molecule compounds in BT474 cells.

[0222] Figure 26 shows the experimental results of the anti-proliferative activity of small molecule compounds in HCC1569 cells.

[0223] Figure 27 shows the experimental results of the antiproliferative activity of the small molecule compound in MV-4-11 cells.

[0224] Figure 28 shows the experimental results of the antiproliferative activity of the small molecule compound in MV-4-11 cells.

[0225] Figure 29 shows the experimental results of the antiproliferative activity of the small molecule compound in MDA-MB-468 cells.

[0226] Figure 30 shows the experimental results of the anti-proliferative activity of N87 cells from ADC samples.

[0227] Figure 31 shows the experimental results of the anti-proliferative activity of N87 cells from ADC samples.

[0228] Figure 32 shows the experimental results of the anti-proliferative activity of N87 cells from the ADC sample.

[0229] Figure 33 shows the experimental results of the anti-proliferative activity of BT474 cells in ADC samples.

[0230] Figure 34 shows the experimental results of the anti-proliferative activity of BT474 cells in ADC samples.

[0231] Figure 35 shows the experimental results of the anti-proliferative activity of SK-OV-3 cells in the ADC sample.

[0232] Figure 36 shows the experimental results of the anti-proliferative activity of ADC samples in MDA-MB-231 cells.

[0233] Figure 37 shows the experimental results of the anti-proliferative activity of ADC samples in HCC1569 cells.

[0234] Figure 38 shows the experimental results of the ADC samples on the Bystander activity of different ratios of effector cells and target cells.

[0235] Figure 39 shows the tumor growth curves of N87 model mice in different treatment groups of ADC samples, where the arrows represent the drug administration time points.

[0236] Figure 40 shows the changes in body weight of N87 model mice bearing tumors in different treatment groups of ADC samples.

[0237] Figure 41 shows the tumor growth curves of SK-OV-3 model mice in different treatment groups of ADC samples, where the arrows represent the drug administration time points.

[0238] Figure 42 shows the changes in body weight of SK-OV-3 model mice in different treatment groups of ADC samples. Detailed Implementation

[0239] Unless otherwise defined, all scientific and technical terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art.

[0240] The term "alkyl" refers to a straight-chain or branched hydrocarbon radical that does not contain unsaturated bonds and is connected to the rest of the molecule by a single bond. Typical alkyl groups contain 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) carbon atoms, preferably 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, n-hexyl, isohexyl, etc. If the alkyl group is substituted with an aryl group, it is referred to as "aralkyl," such as benzyl, diphenylmethyl, or phenethyl. If the alkyl group is substituted with a heterocyclic group, it is referred to as "heterocyclic alkyl." In this invention, CO alkyl refers to H, i.e., C 0-10 Alkyl (or C0-C) 10 Alkyl groups include H and C. 1-10 Alkyl (or C1-C) 10 alkyl).

[0241] The term "alkylene" refers to a hydrocarbon group (divalent alkyl) formed by the loss of two hydrogen atoms from an alkane molecule. It can be straight-chain or branched and is connected to the rest of the molecule by a single bond. Typical alkylene groups described herein have 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) carbon atoms, preferably 1 to 6 carbon atoms, such as methylene (-CH2-), ethylene, propylene, butylene, etc. In this invention, CO alkylene refers to a single bond, i.e., C... 0-10 Alkylene (or C0-C) 10 Alkylenes include single bonds and C bonds. 1-10 Alkylene (or C1-C) 10 (alkylene).

[0242] The term "alkoxy" refers to a substituent formed when the hydrogen in a hydroxyl group is replaced by an alkyl group, such as alkoxy groups containing 1-10 carbon atoms, such as methoxy, ethoxy, propoxy, butoxy, etc.

[0243] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0244] The term "haloalkyl" refers to a group formed by replacing one or more hydrogen atoms in an alkyl group with a halogen atom (such as fluorine, chlorine, bromine or iodine), such as -CHF2, -CH2F, -CF3, -CH2-CF3, -CH2CH2-CF3, -CH2CH2CH2-CF3.

[0245] The term "cycloalkyl" refers to alicyclic hydrocarbons, such as those containing 1 to 4 monocyclic and / or fused rings, containing 3 to 18 carbon atoms, preferably 3 to 10 (e.g., 3, 4, 5, 6, 7, 8, 9, 10) carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or adamantyl.

[0246] The term "aryl" refers to a monocyclic or polycyclic free radical, including polycyclic free radicals containing a monoaryl group and / or a fused aryl group (also referred to herein as "aromatic ring"), such as those containing 1-3 monocyclic or fused rings and 6-18 (e.g., 6, 8, 10, 12, 14, 16, 18) carbon ring atoms, C6-C as described in this invention. 12 The aryl group refers to an aryl group containing 6-12 carbon ring atoms, such as phenyl, naphthyl, biphenyl, indene, etc.

[0247] The term "heterocyclic group" refers to a 3- to 18-membered non-aromatic ring group containing 2 to 17 carbon atoms and 1 to 10 heteroatoms. Heterocyclic groups can be monocyclic, bicyclic, tricyclic, or tetracyclic ring systems, and can include fused, spirocyclic, or bridged ring systems. Heterocyclic groups (also referred to herein as "heterocycles") can be partially saturated (heteroaryl, also referred to herein as "heteroaromatic rings") or fully saturated (heterocyclic alkyl). Suitable heteroaryl groups in the compounds of the present invention contain one, two, or three heteroatoms selected from N, O, S, and P atoms. These heteroaryl groups include, for example, coumarin (including 8-coumarin), quinolinyl (including 8-quinolinyl, isoquinolinyl, pyridinyl, pyrazinyl, pyrazolyl, pyrimidinyl, furanyl, pyrroloyl, thiopheneyl, thiazolyl, isothiazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, imidazoleyl, indolyl, isoyndolyl, indazoleyl, inazinyl, phthalazinyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazolidyl, pyridazinyl, triazinyl, cenolinyl, benzimidazolyl, benzofuranyl, benzofuranyl, benzothiopheneyl, benzothiazolyl, benzooxazolyl, quinazolinyl, quinoxolinyl, naphridinyl, and furanopyridinyl. Suitable heterocyclic alkyl groups in the compounds of the present invention contain one, two, or three heteroatoms selected from N, O, or S atoms. These heterocyclic alkyl groups include, for example, pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, tetrahydrothiophenyl, piperidinyl, morpholinyl, thiomorpholinyl, oxothiohexacyclohexyl, piperazine, aziridine, oxothiohexacyclohexyl, thiohexacyclohexyl, homopiperidinyl, oxopropane, thiopropane, acrylonitrile, oxo-aziridine, diaziridine, etc. Heptyl, triacetyl, 1,2,3,6-tetrahydropyridyl, 2-pyrrolinyl, 3-pyrrolinyl, dihydroindolyl, 2H-pyranyl, 4H-pyranyl, dioxacyclohexyl, 1,3-dioxapentyl, pyrazolinyl, dithiaalkyl, dithiopentyl, dihydropyranyl, dihydrothiophenyl, pyrazolinyl, imidazolinyl, imidazolinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, 3H-indolyl, and quinazinyl. In this invention, for optionally substituted heterocyclic groups, the substituted position can be any suitable carbon atom or heteroatom, for example, for The substitution position of R can be any suitable carbon or nitrogen atom, and it can be, for example...

[0248] The compounds of the present invention also include isotopically labeled forms, that is, compounds distinguished only by the presence of one or more isotopically rich atoms. For example, compounds having existing structures in which at least one hydrogen atom is replaced only by deuterium or tritium, or at least one carbon atom is replaced by carbon rich in 13C or 14C, or at least one nitrogen atom is replaced by nitrogen rich in 15N are all included within the scope of the present invention.

[0249] The term "pharmaceutically acceptable" means that when the molecular basis and the composition comprising it are properly administered to a subject, they do not produce adverse, allergic or other adverse reactions.

[0250] The term "pharmaceutically acceptable salt" includes acid addition salts and base addition salts.

[0251] The term "acid addition salt" includes, but is not limited to, salts derived from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, and phosphonic acid, as well as salts derived from organic acids such as aliphatic monocarboxylic acids and dicarboxylic acids, phenyl-substituted alkanes, hydroxyalkanes, alkanedioic acids, aromatic acids, and aliphatic and aromatic sulfonic acids. Therefore, these salts include, but are not limited to, sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, hydrochlorides, hydrobromates, iodates, acetates, propionates, octanoates, isobutyrates, oxalates, malonates, succinates, octanoates, sebacic acid salts, fumarates, maleates, amygdalinates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, phthalates, benzenesulfonates, toluenesulfonates, phenylacetates, citrates, lactates, maleates, tartrates, and methanesulfonates, as well as salts of amino acids such as arginine salts, gluconates, and galacturonic acids. Acid addition salts can be prepared by contacting a sufficient amount of the desired acid in a conventional manner to form a salt. The free base can be regenerated by contacting the salt with a base, and the free base can be separated in a conventional manner.

[0252] The term "base addition salt" refers to a salt formed with a metal or amine, such as hydroxides of alkali metals and alkaline earth metals, or with an organic amine. Examples of metals used as cations include, but are not limited to, sodium, potassium, magnesium, and calcium. Suitable amines include, but are not limited to, N,N′-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine (1,2-diaminoethane), N-methylglucosamine, and procaine. Base addition salts can be prepared by contacting the free acid form with a sufficient amount of the desired base in a conventional manner to form the salt. The free acid form can be regenerated by contacting the salt form with an acid, and the free acid can be separated in a conventional manner.

[0253] The term "stereoisomer" includes enantiomers, diastereomers, and geometric isomers. Some compounds of the present invention have cyclic hydrocarbon groups that can be substituted on more than one carbon atom; in this case, all their geometric forms, including cis and trans, and mixtures thereof, are within the scope of the present invention.

[0254] The term "solvent" refers to the physical bond between the compound of this invention and one or more solvent molecules. This physical bond includes various degrees of ionic and covalent bonding, including hydrogen bonding. In some cases, the solvate can be isolated, for example when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. Solvents include solution phases and separable solvates. Representative solvates include ethanolides, methanolides, etc.

[0255] The term "prodrug" refers to a Formula I compound that is suitable for administration to patients without excessive toxicity, irritation, or allergic reactions, and is effective for its intended purpose. Prodrugs include acetals, esters, and zwitterionic forms. Prodrugs are converted in the body, such as through hydrolysis in the blood, to yield the parent compound.

[0256] The terms “patient” or “subject”, etc., may be used interchangeably herein to refer to any animal or its cells, whether in vitro or in situ, treated according to the methods described herein. Specifically, the aforementioned animals include mammals, such as rats, mice, guinea pigs, rabbits, dogs, monkeys, or humans, especially humans.

[0257] The term "treatment" refers to the prevention, cure, reversal, reduction, mitigation, minimization, suppression, cessation, and / or cessation of one or more clinical symptoms of a disease after its onset.

[0258] The term "prevention" refers to the treatment taken before a disease develops to avoid, minimize, or prevent the disease from developing or progressing.

[0259] The term "tumor" refers to an abnormal mass of tissue that grows beyond and out of harmony with the growth of normal tissue. Tumors can be "benign" or "malignant," depending on characteristics such as the degree of cell differentiation (including morphology and function), growth rate, local invasion, and metastasis. "Benign tumors" are typically well-differentiated, characterized by slower growth than malignant tumors, and remain confined to their site of origin. Furthermore, benign tumors do not have the ability to infiltrate, invade, or metastasize to distant sites. In some cases, certain "benign" tumors may later develop into malignant tumors, possibly due to additional genetic alterations in a subset of the tumor's proliferative cells, and these tumors are called "precancerous tumors." "Malignant tumors" are typically poorly differentiated (anaplastic) and characterized by rapid growth, accompanied by progressive infiltration, invasion, and destruction of surrounding tissues. Furthermore, malignant tumors often have the ability to metastasize to distant sites.

[0260] The term “cancer” refers to a malignant tumor (Stedman’s Medical Dictionary, 25th ed.; Hensyl ed.; Williams & Wilkins: Philadelphia, 1990).

[0261] The compounds disclosed in this invention can be used as pharmaceuticals (such as targeted protein degraders, ADC drugs) for the prevention or treatment of diseases, such as tumors, including but not limited to: colorectal cancer (e.g., colon cancer, rectal cancer, familial colorectal cancer, hereditary nonpolyposis colorectal cancer), anal cancer, lung cancer (e.g., non-small cell lung cancer, small cell lung cancer), mesothelioma, pancreatic cancer (e.g., pancreatic ductal carcinoma, pancreatic endocrine tumor), pharyngeal cancer, laryngeal cancer, esophageal cancer, gastric cancer (e.g., papillary adenocarcinoma, mucinous adenocarcinoma, adenosquamous carcinoma), duodenal cancer, small bowel cancer, breast cancer (e.g., invasive ductal carcinoma, non-invasive ductal carcinoma, inflammatory breast cancer), ovarian cancer (e.g., ovarian epithelial carcinoma, gonadal extragerminal tumor, ovarian germ cell tumor, low-potency ovarian tumor), testicular tumors, prostate cancer (e.g., hormone-dependent prostate cancer, non-hormone-dependent prostate cancer, castration-resistant prostate cancer), liver cancer (e.g., hepatocellular carcinoma, primary liver cancer, liver cancer). External bile duct cancer), thyroid cancer (e.g., medullary thyroid carcinoma, papillary thyroid carcinoma), kidney cancer (e.g., renal cell carcinoma, transitional cell carcinoma of the renal pelvis and ureter), uterine cancer (e.g., cervical cancer, endometrial cancer, uterine sarcoma), choriocarcinoma of pregnancy, brain tumors (e.g., medulloblastoma, glioma, pineal astrocytoma, pilocytic astrocytoma, diffuse astrocytoma, anaplastic astrocytoma, pituitary adenoma), retinoblastoma, skin cancer (e.g., basal cell carcinoma, malignant melanoma), sarcoma (e.g., rhabdomyosarcoma, leiomyosarcoma, soft tissue sarcoma, spindle cell sarcoma), malignant bone tumors, bladder cancer, hematologic malignancies (e.g., multiple myeloma), lymphoma (e.g., non-Hodgkin lymphoma (NHL), Hodgkin lymphoma), leukemia (e.g., acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL)), chronic myeloproliferative disorders, myelodysplastic syndromes (MDS)).

[0262] The term "Drug to Antibody Ratio" (sometimes written as "Dar" in this document) refers to the average number of target protein degraders attached to each antibody molecule. The conjugates described herein have DAR values ​​from 1 to 10, for example: 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2. 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9 or 10.

[0263] The term "antibody" also refers to a full-length immunoglobulin molecule or the immunoactive portion of a full-length immunoglobulin molecule, i.e., a molecule containing an antigen-binding site that immune-specifically binds to an antigen or a portion thereof that is a target of interest, including but not limited to cancer cells or cells that produce autoantibodies associated with autoimmune diseases. This encompasses naturally occurring or partially or wholly synthetic immunoglobulins and fragments thereof. The term also encompasses any protein having a binding domain homologous to an immunoglobulin binding domain. Antibodies also include polypeptides containing a framework region of an immunoglobulin gene or a fragment thereof that specifically binds to and recognizes an antigen. The use of "antibody" is intended to include complete antibodies, polyclonal, monoclonal, and recombinant antibodies, fragments thereof, and also includes single-chain antibodies, humanized antibodies, mouse antibodies, chimeric antibodies, mouse-human, mouse-primate, primate-human monoclonal antibodies, anti-idiotypic antibodies, and antibody fragments. Antibodies include bispecific and multispecific antibodies, provided they exhibit the desired biological activity or function. In some aspects herein, the biologically active molecule is an antibody or a molecule containing its antigen-binding fragment.

[0264] The term "antibody fragment" contains a portion of a complete antibody, typically its antigen-binding region or variable region.

[0265] The term "connector" refers to the chemical structure in an antibody-drug conjugate (ADC) that links the antibody to the drug payload. In this paper, a connector refers to any chemical part capable of attaching the binding moiety (Bm) to group D in the conjugate of formula (Ⅳ). Attachment to "Bm" can be achieved through chemical or enzymatic conjugation, or a combination of both. Connectors can be classified as either cleavable or non-cleavable.

[0266] The term "cleavable linker" refers to a class of linkers that can be cleaved to release a drug payload, and is sensitive to acid-induced cleavage, light-induced cleavage, bioreduction cleavage, and enzymatic cleavage. In some respects, cleavable linkers can be cleaved by proteases, peptidases, esterases, β-glucuronidases, glycosidases, phosphodiesterases, phosphatases, pyrophosphatases, or lipases. Examples of proteases include, but are not limited to, cathepsin B and VAGP tetrapeptide.

[0267] The term "uncleavable linker" refers to a class of linkers that require endocytosis and lysosomal degradation to release the drug payload, or any chemical part that can stably, covalently attach its binding portion to a new degrading agent and does not fall into the category defined herein as "cleavable linkers." Therefore, uncleavable linkers are essentially resistant to acid-induced cleavage, photo-induced cleavage, bioreductive cleavage, peptidase-induced cleavage, esterase-induced cleavage, and disulfide bond cleavage.

[0268] The term "binding moiety" refers to any molecule that recognizes and binds to cell surface markers or receptors. In some respects, binding moieties bind proteins, not limited to polypeptide moieties. In addition to targeting specific cells, tissues, or sites with novel degraders, binding moieties may also possess certain therapeutic effects, such as antiproliferative (cell-inhibiting and / or cytotoxic) activity against target cells or pathways. In some respects, binding moieties may contain, or be engineered to contain at least one chemically reactive group such as a carboxylic acid, amine, thiol, or chemically reactive amino acid moieties or side chains. In some respects, binding moieties may contain a targeting moiety for a given target cell population that binds to or complexes with cell surface molecules, such as cell surface receptors or antigens. Upon receptor-specific binding or complexation, cells are allowed to take up the targeting moieties or novel degrader conjugates and then internalize them into the cell.

[0269] In this paper, the group "Bm" can be conjugated with more than one targeted protein degrader. For example, "Bm" can be conjugated with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 targeted protein degraders.

[0270] The term "antibody-drug conjugate" (ADC) is a targeted biologic agent in which a varying number of small molecular cytotoxic drugs (SMs) are linked to a monoclonal antibody via a linker. In other words, ADC drugs are typically designed with three main components: an antibody, a linker, and a small molecular cytotoxic drug, combining the high targeting specificity of monoclonal antibodies with the high activity of cytotoxic drugs. In this article, the terms "antibody-drug conjugate," "antibody-drug conjugate," "ADC drug," "ADC sample," "antibody-drug conjugate," and "conjugate" are used interchangeably. Similarly, the terms "small molecular cytotoxic drug," "cytotoxic drug," "small molecular cytotoxin," "cytotoxin," "payload," "load," and "drug load" are used interchangeably.

[0271] In this article, the terms "targeted protein degrader", "degrader", "inhibitor", "small molecule compound", and "compound" can be used interchangeably.

[0272] All publications, patents, and published patent specifications cited in this article are incorporated herein in their entirety through citation.

[0273] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0274] The “Dxd (Exatecan derivative for ADC)” used in the following examples is an effective topoisomerase I inhibitor that can be used as a payload for the antibody-drug conjugate DS-8201 targeting HER2. Its structure is as follows: SMol006 is a molecular adhesive degrader. Referring to patent document WO2021198965, its structure is as follows:

[0275] Example 1: Synthesis of Compound 1

[0276] 1. Synthesis of compounds 1-2

[0277] To a solution of compound 1-1 (10.00 g, 55.2 mmol, 1.0 eq.) in concentrated sulfuric acid (30 mL), 1,3-dichloro-5,5-dimethylhydantoin (13.06 g, 66.2 mmol, 1.2 eq.) was added. The mixture was stirred at 80 °C for 12 hours. TLC (n-heptane / EA = 1:1) showed that the reaction was complete. The mixture was poured into ice water (200 mL), filtered, and dried under reduced pressure to give compound 1-2 (16.10 g, crude product), which was a white solid. The crude product could be used in the next step without further purification.

[0278] 1 H NMR (400MHz, DMSO-d6): δ14.0(s,1H),8.43–8.31(m,2H),2.60(s,3H).

[0279] 2. Synthesis of compounds 1-3

[0280] Morpholine (606.15 mg, 6.96 mmol, 1.0 eq.), DIPEA (1.80 g, 13.92 mmol, 2.0 eq.), and HATU (3.17 g, 8.35 mmol, 1.2 eq.) were added to a DCM (30 mL) solution of compounds 1-2 (1.5 g, 6.96 mmol, 1.0 eq.). The mixture was stirred at 27 °C for 2 hours. TLC showed that the reaction was complete. The mixture was washed three times with water (20 mL), the organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (DCM / EA = 1 / 0 to 0 / 1) to give compounds 1-3 (1.74 g) as a white solid.

[0281] 1 H NMR (400MHz, DMSO-d6): δ8.46(s,1H),8.25(s,1H),4.03–3.65(m,6H),3.52(d,J=54.2Hz,2H),2.65(s,3H).

[0282] 3. Synthesis of compounds 1-4

[0283] At 0 °C, a borane dimethyl sulfide complex (1.49 mL, 14.93 mmol, 2.5 eq.) was added dropwise to a THF (85 mL) solution of compounds 1-3 (1.7 g, 5.97 mmol, 1.0 eq.). The mixture was stirred at 70 °C for 2 h. TLC showed that the reaction was complete. The mixture was quenched with methanol (4.5 mL), refluxed for 1 h, and concentrated under reduced pressure to give the residue. The residue was purified by silica gel column chromatography (DCM / EA = 1 / 0 to 0 / 1) to give compounds 1-4 (1.4 g, 86.6% yield) as a white solid.

[0284] 1 H NMR (400MHz, DMSO-d6): δ8.19(d,J=2.5Hz,1H),8.12(d,J=2.5Hz,1H),3.61(s,2 H),3.58(t,J=4.6Hz,4H),2.47(s,3H),2.41(t,J=4.6Hz,4H).LC-MS:-271[M+1].

[0285] 4. Synthesis of compounds 1-5

[0286] To a solution of compounds 1-4 (1.2 g, 4.43 mmol, 1.0 eq.) in ethanol (30 mL) and H₂O (6 mL), NH₄Cl (711.34 mg, 13.30 mmol, 3.0 eq.) and Fe (1.24 g, 22.16 mmol, 5.0 eq.) was added. The mixture was stirred at 70 °C for 2 hours. TLC (DCM / EA = 1:1) showed that the reaction was complete. The mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to give the residue. The residue was purified by silica gel column chromatography (DCM / EA = 1 / 0 to 0 / 1) to give compounds 1-5 (600 mg, 56.2% yield) as a yellow solid.

[0287] 1 H NMR (400MHz, DMSO-d6): δ6.53(d,J=2.4Hz,1H),6.46(s,1H),5.09(s,2H),3.55(d ,J=9.3Hz,4H),3.29(s,2H),2.33(d,J=4.8Hz,4H),2.17(s,3H).LCMS:241[M+1].

[0288] 5. Synthesis of Compound 1

[0289] At 0 °C, triphosgene (44.38 mg, 149.55 μmol, 0.36 eq.) was added to a THF (10 mL) solution of compounds 1-5 (100 mg, 415.41 μmol, 1.0 eq.). The mixture was stirred at 25 °C for 2 hours. The sample was quenched with MeOH and monitored by TLC. TLC (DCM / EA = 1 / 1) showed that the reaction was complete.

[0290] 3-[5-(aminomethyl)-1-oxoisoindolin-2-yl]piperidine-2,6-dione (113.52 mg, 415.41 μmol, 1.0 eq.), DMF (15 mL), and TEA (420.35 mg, 4.15 mmol, 10.0 eq.) were mixed and stirred at 25 °C for 30 min, and then added to the freshly prepared isocyanate. The resulting mixture was stirred at 25 °C for 1.5 h. TLC (DCM / EA = 1 / 1) showed that the reaction was complete. The mixture was quenched with water (20 mL) and extracted with ethyl acetate (3 × 20 mL). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by preparative HPLC in HCl buffer to give compound 1 (31 mg, 12.9% yield) as a white solid.

[0291] 1 H NMR (400MHz, DMSO-d6): δ11.00(s,1H),10.02(s,1H),9.07(s,1H),7.74–7.66(m,2H),7.57 –7.42(m,3H),7.15(t,J=6.1Hz,1H),5.11(dd,J=13.3,5.1Hz,1H),4.49–4.24(m,6H),3.93( d,J=12.5Hz,2H),3.71(t,J=12.1Hz,2H),3.24(dt,J=34.7,11.7Hz,4H),2.91(ddd,J=17.2 ,13.6,5.4Hz,1H),2.60(d,J=17.0Hz,1H),2.36(s,4H),1.99(dtd,J=12.7,5.4,2.2Hz,1H).

[0292] Example 2: Synthesis of Compound 2

[0293] 1. Synthesis of Compound 2-1

[0294] At 27 °C, DIEA (3.60 g, 27.8 mmol, 2.00 eq.) was added dropwise to a DCM (60 mL) solution of compounds 1-2 (3.00 g, 13.9 mmol, 1.00 eq.) and stirred for 5 minutes. Then, HATU (6.35 g, 16.7 mmol, 1.20 eq.) was added to the mixture and stirred for 2 hours. TLC (n-heptane / EA = 1 / 1) showed complete reaction. The mixture was poured into ice water (120 mL) and extracted with DCM (2 × 120 mL). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give compound 2-1 (7.7 g, crude), which was a yellow oil.

[0295] LCMS: 286 [M+H-100]

[0296] 2. Synthesis of compound 2-2

[0297] At 0 °C, BH3·DMS (1.40 mL, 14.0 mmol, 2.00 eq.) was added dropwise to a THF (99.2 mL) solution of compound 2-1 (2.70 g, 7.00 mmol, 1.00 eq.). The mixture was stirred at 70 °C for 1.5 h. TLC (n-heptane / EA = 1 / 1) showed that the reaction was complete. The mixture was cooled to 25 °C and quenched with methanol (5 mL). The solution was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM / EA = 1 / 0 to 0 / 1) to give compound 2-2 (2.30 g, 88% yield).

[0298] 1 H NMR (400MHz, CDCl3): δppm 8.13(d,J=2.4Hz,1H),8.11-8.04(m,1H),3.79-3.69(m,2H),3.33(dt,J=24.4,6.8H z,2H),2.80(s,3H),2.61-2.49(m,2H),2.45(s,3H),2.26(s,3H),1.50-1.33(m,9H).

[0299] 3. Synthesis of compounds 2-3

[0300] Fe (1.65 g, 29.6 mmol, 5.00 eq.) was added fractionally to a solution of compound 2-2 (2.20 g, 5.92 mmol, 1.00 eq.) and NH4Cl (1.58 g, 29.6 mmol, 5.00 eq.) in ethanol (130 mL) and H2O (32 mL) at 25 °C. The mixture was stirred at 70 °C for 2 hours. TLC (DCM / EA = 1 / 1) showed that the reaction was complete. The mixture was cooled to 25 °C, filtered with diatomaceous earth, and washed with ethyl acetate (20 mL). The filtrate was separated, and the aqueous phase was extracted with ethyl acetate (2 × 20 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by silica gel column chromatography (DCM / EA = 1 / 0 to 0 / 1) to give compound 2-3 (350 mg, 17% yield).

[0301] 1 H NMR (400MHz, CDCl3): δppm 6.62(s,2H),3.67(d,J=41.2Hz,2H),3.34(d,J=39.6Hz,4H),2.80(d,J=9.6 Hz,3H),2.59-2.38(m,2H),2.33-2.12(m,6H),1.43(s,9H).LCMS:342[M+H].

[0302] 4. Synthesis of Compound 2

[0303] Triphosgene (109.37 mg, 0.368 mmol, 0.36 eq.) was added to a THF (10 mL) solution of compounds 2-3 (350 mg, 1.02 mmol, 1.00 eq.). The solution was stirred at 25 °C for 2 h to prepare the corresponding isocyanates. The samples were quenched with MeOH and monitored by TLC. TLC (DCM / EA = 1 / 1) showed that the reaction was complete.

[0304] At 0 °C, TEA (1.04 g, 10.24 mmol, 10 eq.) was added to a DMF (3.5 mL) solution of 3-[5-(aminomethyl)-1-oxoisoindolin-2-yl]piperidine-2,6-dione hydrochloride (280 mg, 903 μmol, 0.88 eq.). The mixture was stirred at 25 °C for 30 min, and then added to the above isocyanate solution. The resulting mixture was stirred for 1.5 h. TLC (DCM / EA = 1 / 1) showed that the reaction was complete. The mixture was quenched with water (20 mL) and extracted with ethyl acetate (3 × 20 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by preparative HPLC to give compound 2 protected by Boc as a white solid. The solid was dissolved in dioxane (10 mL), and HCl / dioxane (4 M, 1 mL) was added to the solution. The resulting solution was stirred at 25°C for 2 hours. LCMS showed that the reaction was complete. The solution was concentrated under reduced pressure, purified by preparative HPLC, and lyophilized to give compound 2 (HCl salt, 55.3 mg), which was a white solid.

[0305] 1 H NMR (400MHz, DMSO-d6): δppm 7.78-7.66(m,2H),7.57-7.39(m,3H),5.06(dd,J=13.2,5.2Hz,1H),4.57-4.20(m,6H),3.43(dt,J=30.8,6.4Hz,4H),2.87( ddd,J=17.2,13.6,5.2Hz,1H),2.78(s,3H),2.58(s,4H),2.35(s,4H),2.00(ddd,J=10.4,5.2,3.2Hz,1H).LCMS:541[M+H].

[0306] Example 3: Synthesis of Compound 3

[0307] 1. Synthesis of compound 3-1

[0308] To a DCM (90 mL) solution of compounds 1-2 (4.00 g, 18.6 mmol, 1.0 eq.) and N-Boc-piperazine (3.46 g, 18.6 mmol, 1.0 eq.), DIEA (4.80 g, 37.1 mmol, 2.0 eq.) and HATU (8.47 g, 22.3 mmol, 1.20 eq.) were added. The mixture was stirred at 27 °C for 2 h. TLC (n-heptane / EA = 1 / 1) showed that the reaction was complete. The mixture was poured into ice water (120 mL) and separated in a funnel. The aqueous phase was extracted with DCM (2 × 120 mL). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give a crude oil. Purification by silica gel column chromatography (DCM / EA = 1 / 0 to 0 / 1) gave compound 3-1 (3.3 g, 46% yield) as a yellow oil.

[0309] 1 H NMR (400MHz, CDCl3): δ8.27(d,J=2.4Hz,1H),7.96(d,J=2.4Hz,1H),3.93-3.74(m,2H),3.55(t ,J=5.2Hz,2H),3.38(s,2H),3.19(s,2H),2.43(s,3H),1.48-1.41(m,9H).LCMS:286[M+H-tBu].

[0310] 2. Synthesis of compound 3-2

[0311] At 0 °C, BH3·DMS (1.67 mL, 16.7 mmol, 2.00 eq.) was added dropwise to a THF (4.05 mL) solution of compound 3-1 (3.20 g, 8.34 mmol, 1.00 eq.). The mixture was stirred at 70 °C for 1.5 h. TLC (n-heptane / EA = 1 / 1) showed that the reaction was complete. The mixture was cooled to 25 °C, quenched with methanol (10 mL), and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM / EA = 1 / 0 to 0 / 1) to give compound 3-2 (2.00 g, 64% yield).

[0312] 1 H NMR (400MHz, CDCl3): δppm 8.16(d,J=2.4Hz,1H),8.08(d,J=2.4Hz,1H),3.54(s,2H),3.43(t,J=5.2Hz,4H),2.48(s,3H),2.42(d,J=5.6Hz,4H),1.46(s,9H).

[0313] 3. Synthesis of compound 3-3

[0314] Fe (574 mg, 10.28 mmol, 2.00 eq.) was added fractionally to a solution of compound 3-2 (1.90 g, 5.14 mmol, 1.00 eq.) and NH4Cl (550 mg, 10.28 mmol, 2.00 eq.) in ethanol (30 mL) and H2O (6 mL) at 25 °C. The mixture was stirred at 70 °C for 2 hours. TLC (DCM / EA = 1 / 1) showed that the reaction was complete. The mixture was cooled to 25 °C, filtered with diatomaceous earth, and washed with ethyl acetate (20 mL). The filtrate was separated, and the aqueous phase was extracted with ethyl acetate (2 × 20 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography to give compound 3-3 (1.00 g, 57.5% yield).

[0315] 1 H NMR (400MHz, DMSO-d6): δppm 6.50(d,J=2.4Hz,1H),6.42(d,J=2.4Hz,1H),5.07(s,2H),3.31(s,4H),3.26(s,2H),2.25(d,J=4.0Hz,4H),2.12(s,3H),1.35(s,9H).

[0316] 4. Synthesis of compounds 3-4

[0317] Triphosgene (94.3 mg, 0.318 mmol, 0.36 eq.) was added to a THF (10 mL) solution of compound 3-3 (300 mg, 0.883 mmol, 1.00 eq.). The solution was stirred at 25 °C for 2 h to prepare the corresponding isocyanate. The sample was quenched with MeOH and monitored by TLC. TLC (DCM / EA = 1 / 1) showed that the reaction was complete.

[0318] At 0 °C, TEA (897 mg, 8.83 mmol, 10 eq.) was added to a DMF (3.5 mL) solution of 3-[5-(aminomethyl)-1-oxoisoindolin-2-yl]piperidine-2,6-dione hydrochloride (273 mg, 883 μmol, 1.00 eq.). The mixture was stirred at 25 °C for 30 min, and then added to the above isocyanate solution. The resulting mixture was stirred for 1.5 h. TLC (DCM / EA = 1 / 1) showed that the reaction was complete. The mixture was quenched with water (25 mL) and extracted with ethyl acetate (3 × 25 mL). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by preparative HPLC and lyophilized to give compounds 3-4 (250 mg, 44% yield) as a white solid.

[0319] LCMS: 639 [M+H].

[0320] 5. Synthesis of Compound 3

[0321] At 25 °C, HCl / dioxane (4 M, 1 mL) was added dropwise to a solution of compounds 3-4 (30.0 mg, 46.9 μmol, 1.00 eq.) in dioxane (1 mL). The resulting solution was stirred at 25 °C for 5 hours. LC-MS showed that the reaction was complete. The solution was concentrated under reduced pressure, and the residue was purified by preparative HPLC and lyophilized to give compound 3 (HCl salt, 20.0 mg) as a white solid.

[0322] 1 H NMR (400MHz, DMSO-d6): δppm 11.00(s,1H),9.06(s,1H),8.31(s,1H),7.69(d,J=8.0Hz,1H),7.63(d,J=2.4Hz,1H ),7.51(s,1H),7.44(dd,J=8.0,1.6Hz,1H),7.16(d,J=2.4Hz,1H),7.10(t,J=6.0Hz, 1H),5.11(dd,J=13.2,5.2Hz,1H),4.56-4.20(m,4H),3.01-2.73(m,5H),2.72-2.53( m, 2H), 2.40 (tt, J = 8.8, 3.6Hz, 5H), 2.25 (s, 3H), 2.00-1.97 (m, 1H), LCMS: 539 [M+H].

[0323] Example 4: Synthesis of Compound 4

[0324] 1. Synthesis of compound 4-1

[0325] At 26 °C, NH₄Cl (2.48 g, 46.38 mmol, 2.0 eq.), DIPEA (8.99 g, 69.58 mmol, 3.0 eq.), and HATU (10.58 g, 27.83 mmol, 1.2 eq.) were added to a DCM (50 mL) solution of compounds 1-2 (5.00 g, 23.19 mmol, 1.0 eq.). The mixture was stirred for 2 hours. TLC showed that the reaction was complete. The mixture was diluted with DCM (200 mL) and washed with water (3 × 100 mL). The organic layer was dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by silica gel column chromatography (DCM / EA = 1 / 1) to give compound 4-1 (2 g, 40.19% yield) as a yellow solid. 1H NMR (400MHz, DMSO-d6): δ8.35 (d, J = 2.4Hz, 1H), 8.20–8.03 (m, 2H), 7.86 (s, 1H), 2.47 (s, 3H).

[0326] 2. Synthesis of compound 4-2

[0327] At 0 °C, a borane dimethyl sulfide complex (10 M, 1.77 mL, 17.71 mmol, 2.0 eq.) was added to a THF (20 mL) solution of compound 4-1 (1.9 g, 8.85 mmol, 1.0 eq.). The reaction mixture was stirred at 60 °C for 2 h. LCMS showed that the reaction was complete. The mixture was quenched with methanol (40.0 mL), refluxed for 2 h, and concentrated under reduced pressure to give compound 4-2 (1.5 g, crude product), which was a white solid. The crude product could be used for the next step without further purification. LCMS: 201 [M+1].

[0328] 3. Synthesis of compound 4-3

[0329] Compound 4-2 (1.5 g, 7.48 mmol, 1.0 eq.), 2-((tert-butyldiphenylsilyl)oxy)acetic acid (2.35 g, 7.48 mmol, 1.0 eq.), DIEA (2.90 g, 22.43 mmol, 3.0 eq.), and HATU (3.41 g, 8.97 mmol, 1.2 eq.) were added to a DMF (15 mL) solution and stirred for 2 hours. LC-MS showed that the reaction was complete. The mixture was quenched with water (100 mL) and extracted with EtOAc (3 × 100 mL). The combined organic layers were washed with brine (200 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by silica gel column chromatography (DCM / EA = 1 / 1) to give compound 4-3 (450 mg, 12.11% yield, two steps), which was a yellow oil. 1 H NMR (400MHz, DMSO-d6): δ8.41(t,J=6.1Hz,1H),8.23(d,J=2.4Hz,1H),8.07(d,J=2.4Hz,1H),7.65(dt ,J=6.7,1.5Hz,4H),7.49–7.42(m,6H),4.51(d,J=6.1Hz,2H),4.12(s,2H),2.46(s,3H),1.04(s,9H).

[0330] 4. Synthesis of compound 4-4

[0331] To a solution of compound 4-3 (450 mg, 905.33 μmol, 1.0 eq.) in ethanol (20 mL) and H₂O (4 mL), NH₄Cl (242.14 mg, 4.53 mmol, 5.0 eq.) and Fe (252.79 mg, 4.53 mmol, 5.0 eq.) were added. The mixture was stirred at 70 °C for 2 hours. TLC showed that the reaction was complete. The mixture was filtered through diatomaceous earth, concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (DCM / EA = 1 / 0 to 0 / 1) to give compound 4-4 (250 mg, 59.12% yield) as a yellow oil. 1 H NMR (400MHz, DMSO-d6): δ7.88(s,1H),7.70–7.57(m,4H),7.44(dt,J=14.5,7.3Hz,6H),6.56(d,J=2.3Hz ,1H),6.45(d,J=2.4Hz,1H),5.11(s,2H),4.23(d,J=5.9Hz,2H),4.08(s,2H),2.12(s,3H),1.01(s,9H).

[0332] 5. Synthesis of compounds 4-5

[0333] Triphosgene (22.24 mg, 74.93 μmol, 0.35 eq.) was added to a THF (5 mL) solution of compound 4-4 (100 mg, 214.10 μmol, 1.0 eq.). The mixture was stirred at 25 °C for 1 h. The sample was quenched with MeOH and monitored by TLC. TLC (DCM / EA = 1 / 1) showed that the reaction was complete.

[0334] 3-[5-(aminomethyl)-1-oxoisoindolin-2-yl]piperidin-2,6-dione (66.32 mg, 214.10 μmol, 1.0 eq.) and TEA (216.6 mg, 2141.0 μmol, 10 eq.) were added to a DMF solution (5 mL) and stirred for 30 minutes, then added to the freshly prepared isocyanate. The resulting mixture was stirred at 25 °C for 1 hour. TLC (DCM / EA = 1 / 1) showed that the reaction was complete. The mixture was quenched with water (20 mL) and extracted with ethyl acetate (3 × 20 mL). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by silica gel column chromatography (DCM / EA = 2 / 1) to give compounds 4-5 (100 mg, 60.95% yield) as a yellow solid.

[0335] 6. Synthesis of Compound 4

[0336] At 0 °C, a THF solution of TBAF (1 M, 0.391 mL, 391.47 μmol, 3.0 eq.) was added to a THF solution of compounds 4-5 (100 mg, 130.49 μmol, 1.0 eq.) in 5 mL. The mixture was stirred at 25 °C for 2 hours. TLC showed that the reaction was complete. The mixture was added to water (20 mL) and extracted with DCM (3 × 10 mL). The organic layer was dried over Na₂SO₄, filtered, and the filtrate was concentrated to give the residue. The residue was purified twice by preparative HPLC under acidic conditions to give compound 4 (7.6 mg, 11% yield) as a white solid. 1 H NMR (400MHz, Methanol-d4): δ7.76(d,J=7.8Hz,1H),7.57–7.45(m,3H),7.15(d,J=2.2Hz,1H),5.13(dd,J=13.2,5.3Hz,1 H),4.54–4.40(m,6H),4.03(s,2H),2.95–2.72(m,2H),2.48(dd,J=13.3,4.7Hz,1H),2.29(s,3H),2.16(d,J=13.1Hz,1H).

[0337] Example 5: Synthesis of Compound 5

[0338] 1. Synthesis of compounds 1-2

[0339] 1,3-Dichloro-5,5-dimethylhydantoin (13.05 g, 66.25 mmol, 1 eq.) was added to a solution of 2-methyl-5-nitrobenzoic acid (10 g, 55.2 mmol, 1 eq.) in H₂SO₄ (30 mL). The mixture was stirred at 80 °C for 16 hours. TLC showed that the reaction was complete. The mixture was poured into ice water (100 mL), and a pale yellow solid precipitated. The resulting mixture was filtered, the filter cake was washed with water, and dried under reduced pressure to give compounds 1-2 (7.5 g, purity determination: 80%) as a grayish-white solid.

[0340] 1 H NMR(400MHz, DMSO-d6): δ8.41(s,2H),2.63(s,3H).LCMS:214.10[MH] - .

[0341] 2. Synthesis of Compound 5-1

[0342] To a DCM (30 mL) solution of compounds 1-2 (3.60 g, 16.70 mmol, 1 eq.), 4-aminotetrahydropyran (1.69 g, 16.70 mmol, 1 eq.), DIEA (4.32 g, 33.40 mmol, 2 eq.), and HATU (7.62 g, 20.04 mmol, 1.2 eq.) were added. The mixture was stirred at 25 °C for 3 hours. TLC showed that the reaction was complete. The reaction solution was concentrated under reduced pressure to give a crude product. The crude product was ground with MeOH (10 mL) to give compound 5-1 (3.3 g, 66.1% yield), which was a white solid.

[0343] 1 H NMR (400MHz, DMSO-d6): δ8.62(d,J=7.4Hz,1H),8.35(d,J=2.2Hz,1H),8.06(d,J=2.2Hz,1H),4.06–3 .94(m,1H),3.94–3.81(m,2H),3.48–3.36(m,2H),2.43(s,3H),1.90–1.76(m,2H),1.60–1.43(m,2H).

[0344] 3. Synthesis of compound 5-2

[0345] Compound 5-1 (2 g, 6.70 mmol, 1 eq.) was dissolved in THF (20 mL) and cooled to 0–10 °C in an ice bath. Under N2 protection, a borane dimethyl sulfide complex (10 M, 2.68 mL, 26.78 mmol, 4 eq.) was added dropwise to the above solution. After addition, the mixture was stirred at 0 °C for 10 min, then the temperature was raised to 70 °C and stirred for 12 h. TLC showed the reaction was complete. The reaction was cooled to 0–10 °C in an ice bath, then 10 mL of methanol was added dropwise and stirred at 25 °C for 1 h. 3.2 mL of a 4 M HCl solution of 1,4-dioxane was added to the mixture in an ice bath and stirred at 25 °C for 30 min. The resulting mixture was filtered to give a crude product, which was then ground with MTBE to obtain the HCl salt form of the reduced product compound 5-2 (2.5 g), which could be used directly in the next step. LCMS: 285.4 [M+H] + .

[0346] 4. Synthesis of compound 5-3

[0347] Di-tert-butyl dicarbonate (1.97 g, 11.33 mmol, 1.5 eq.) was added to a DCM (40 mL) solution of compound 5-2 (2.15 g, 7.55 mmol, 1 eq.) and TEA (2.29 g, 22.65 mmol, 3 eq.). The mixture was stirred at 25 °C for 16 hours. TLC showed that the reaction was complete. The mixture was concentrated under reduced pressure to give a crude product, which was further purified by rapid chromatography using Hep:EA = 5:1 elution to give compound 5-3 (0.97 g, crude product), which could be used directly in the next step. LCMS: 328.08 [M+H-56] + .

[0348] 5. Synthesis of compound 5-4

[0349] Iron powder (655.69 mg, 11.74 mmol, 5 eq.) was added to a solution of compound 5-3 (903.73 mg, 2.35 mmol, 1 eq.) and NH4Cl (376.83 mg, 7.04 mmol, 3 eq.) in ethanol (45 mL) and H2O (9 mL) at 25 °C. The mixture was stirred at 80 °C for 6 hours. TLC showed that the reaction was complete. The mixture was cooled to room temperature and filtered. The filter cake was washed with EtOH, and the filtrate was concentrated under reduced pressure to give a crude product, which was purified by rapid chromatography with elution using Hep:EA = 5:1 to give compound 5-4 (475.1 mg, 57.01% yield) as a yellow solid. 1 H NMR (400MHz, CDCl3): δ6.62 (d, J = 2.0Hz, 1H), 6.38 (s, 1H), 4.27 (s, 2H), 4. 00–3.89(m,2H),3.46–3.32(m,2H),2.20(s,3H),1.64(s,5H),1.40(s,9H).

[0350] 6. Synthesis of compound 5-5

[0351] Triphosgene (99.51 mg, 431.13 μmol, 0.34 eq.) was added dropwise to a THF (4 mL) solution of compound 5-4- (450 mg, 1.26 mmol, 1 eq.) at 25 °C. The mixture was stirred at 25 °C for 3 hours. The reaction was monitored by TLC, which showed that the STM was almost completely consumed. The resulting mixture was used directly in the next step.

[0352] The freshly prepared isocyanate was added to a solution of 3-[5-(aminomethyl)-1-oxoisoindolin-2-yl]piperidine-2,6-dione hydrochloride (392.7 mg, 1.26 mmol, 1 eq.) and TEA (384.94 mg, 3.80 mmol, 3 eq.) in DMF (4 mL). The mixture was stirred at 25 °C for 16 hours. LC-MS showed that the reaction was complete. The reaction mixture was added to water (20 mL) and extracted with EA (3 × 20 mL). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give a crude product, which was purified by rapid chromatography with elution using Hep:EA = 1:1 to give compound 5-5 (482 mg, 58.2%) as a white solid. LC-MS: 598.4 [M + H - 54] + .

[0353] 7. Synthesis of Compound 5

[0354] A solution of 1,4-dioxane in 4M HCl (2 mL) was added to a solution of compound 5-5 (415 mg, 636.33 μmol, 1 eq.). The resulting mixture was stirred at 25 °C for 3 hours. TLC showed that the reaction was complete. The resulting solution was concentrated under vacuum to give compound 5 (406 mg), which could be used directly in the next step. LCMS: 554.4 [M+H] + .

[0355] Example 6: Synthesis of LP01

[0356] Compound L1 (49.1 mg, 66.5 μmol, 1.20 eq.) was added to a DMF (2 mL) solution of compound 2 (30.0 mg, 55.4 μmol, 1.00 eq.) at 25 °C, and stirred for 5 min. Then, HOBT (10.6 mg, 55.4 μmol, 1.00 eq.), pyridine (2 mL), and DIPEA (7.17 mg, 55.4 μmol, 1.00 eq.) were added to the mixture, and the mixture was stirred at 25 °C for 6 h. LC-MS showed the reaction was complete. The mixture was quenched with water (20 mL) and extracted with DCM (3 × 20 mL). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by preparative HPLC (formic acid buffer) to give LP01 (25.1 mg) as a white solid.

[0357] 1H NMR (400MHz, DMSO-d6): δppm 10.99(s,1H),9.99(s,1H),8.79(s,1H),8.09(d,J=7.6Hz,1H),7.81(d,J =8.4Hz,1H),7.69(d,J=7.6Hz,1H),7.65-7.54(m,3H),7.51(s,1H),7.44( d,J=8.4Hz,1H),7.25(dd,J=22.4,8.0Hz,2H),7.10(d,J=2.4Hz,1H),7.00 (s,2H),6.82(s,1H),5.98(t,J=6.0Hz,1H),5.41(s,2H),5.11(dd,J=13.2 ,5.2Hz,1H),4.93(d,J=22.0Hz,2H),4.48-4.27(m,5H),4.19(t,J=7.6Hz ,1H),3.42-3.35(m,6H),2.96(td,J=13.2,6.4Hz,2H),2.75(d,J=6.0Hz,3 H),2.64-2.59(m,1H),2.47-2.35(m,1H),2.23-1.90(m,10H),1.75-1.29( m,10H),1.19(td,J=14.4,12.0,6.4Hz,3H),0.84(dd,J=12.4,6.8Hz,6H).

[0358] Example 7: Synthesis of LP02

[0359] Compound L1 (57.5 mg, 77.9 μmol, 1.20 eq.) was added to a DMF (2 mL) solution of compound 3 (35.0 mg, 64.9 μmol, 1.00 eq.) at 25 °C, and the mixture was stirred for 5 min. Then, HOBT (52.6 mg, 390 μmol, 6.00 eq.), pyridine (30.8 mg, 390 μmol, 6.00 eq.), and DIPEA (50.4 mg, 390 μmol, 6.00 eq.) were added to the mixture, and the mixture was stirred at 25 °C for 6 h. LC-MS showed that the reaction was complete. The mixture was quenched with water (20 mL) and extracted with ethyl acetate (3 × 20 mL). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by preparative HPLC (formic acid buffer) to give LPO₂ (14.1 mg) as a white solid.

[0360] 1H NMR (400MHz, DMSO-d6): δppm 10.98(s,1H),10.00(s,1H),8.86-8.76(m,1H),8.09(d,J=7.6Hz,1H),7 .81(d,J=8.8Hz,1H),7.72-7.64(m,2H),7.61-7.56(m,2H),7.51(s,1H) ,7.44(dd,J=8.0,1.6Hz,1H),7.32-7.26(m,2H),7.10(d,J=2.4Hz,1H), 7.00(s,2H),6.82(d,J=6.0Hz,1H),5.98(t,J=6.0Hz,1H),5.42(s,2H),5 .11(dd,J=13.2,5.2Hz,1H),5.00(s,2H),4.50-4.27(m,5H),4.19(dd,J=8.8,6.8Hz,1H),3.40(d,J=8.4Hz,5H),3.06-2.87(m,3H),2.64-2.55( m,2H),2.42-2.34(m,4H),2.26(s,3H),2.20-1.90(m,5H),1.77-1.29(m ,9H),1.25-1.13(m,3H),0.84(dd,J=12.8,6.8Hz,6H).LCMS:1138[M+H].

[0361] Example 8: Synthesis of LP03

[0362] 1. Synthesis of compound 4-1

[0363] At 0 °C, oxalyl chloride (1.77 g, 13.92 mmol, 1.5 eq.) and DMF (67.81 mg, 927.69 μmol, 0.1 eq.) were added to a DCM (40 mL) solution of compounds 1-2 (2 g, 9.28 mmol, 1.0 eq.). The mixture was stirred at 25 °C for 3 h. After sampling, the mixture was quenched with MeOH and monitored by TLC. TLC showed that the reaction was complete. The mixture was concentrated to obtain fresh acyl chloride.

[0364] In a 100 mL three-necked round-bottom flask, ammonia solution (1.58 g, 92.80 mmol, 10 eq) was stirred in 8 mL of DCM and cooled in an ice bath for 5 minutes. The freshly prepared acyl chloride was then dissolved in 12 mL of DCM and added dropwise to the ammonia solution. The reaction mixture was stirred at 25 °C for 4 hours. TLC showed the reaction was complete. The suspension was filtered, and the solid was collected to give compound 4-1 (1.33 g, yield 66.68%) as a white solid.

[0365] 1 H NMR: 400MHz, DMSO-d6δ8.31(d,J=2.5Hz,1H),8.19–7.99(m,2H),7.80(s,1H),2.45(s,3H).LCMS:215.10[M+H] + .

[0366] 2. Synthesis of Compound 6-1

[0367] In a 50 mL three-necked round-bottom flask, compound 4-1 (0.5 g, 2.33 mmol, 1.0 eq) was dissolved in 5 mL of THF and cooled to 0-10 °C in an ice bath. Under nitrogen protection, a dimethyl borohydride sulfide complex (10 M, 0.932 mL, 9.32 mmol, 4.0 eq) was added dropwise. After the addition was complete, the mixture was stirred at 0 °C for 10 min, then the temperature was raised to 70 °C and maintained for 4 h. TLC showed that the reaction was complete. The mixture was quenched at 0 °C with 4 M hydrochloric acid in 1,4-dioxane (2 mL) and stirred at 25 °C for 30 min. After filtration, the filter cake was treated with MTBE to give compound 6-1a (0.227 g), as its hydrochloride salt.

[0368] LCMS:201.20[M+H] + .

[0369] In a 40 mL test tube, compound 6-1a (0.227 g, 1.13 mmol, 1.0 eq) and TEA (343.48 mg, 3.39 mmol, 3.0 eq) were dissolved in 4 mL of DCM, followed by the addition of Boc₂O (295.72 mg, 1.70 mmol, 1.5 eq). After the addition was complete, the mixture was stirred at 25 °C for 4 hours. TLC showed that the reaction was complete. The reaction was quenched by adding 5 mL of water, and the organic phase was then separated. The aqueous phase was extracted three times with 5 mL of EA. The combined organic phases were dried over Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. Rapid chromatographic purification was performed, eluting with Hep:EA at a ratio of 2:1, to give compound 6-1 (0.233 g, yield of 33.2% in two steps), which was a pale yellow oil with a purity of 93.55%.

[0370] 1 H NMR: 400MHz, CDCl3δ8.17(d,J=2.4Hz,1H),8.10–7.99(m,1H),4.93(s,1H),4.40(d,J=6.3Hz,2H),2.45(s,4H),1.48(s,9H).LCMS:318.20[M+NH3+H].

[0371] 3. Synthesis of Compound 6-2

[0372] Compound 6-1 (1 g, 3.33 mmol, 1.0 eq) and NH4Cl (533.60 mg, 9.98 mmol, 3.0 eq) were dissolved in EtOH (50 mL) and H2O (10 mL), and Fe (928.47 mg, 16.63 mmol, 5.0 eq) was added at 25 °C. The mixture was stirred at 80 °C for 16 hours. TLC showed that the reaction was complete. The mixture was cooled to room temperature and filtered. The filter cake was washed with EtOH. The filtrate was concentrated under reduced pressure to give a crude product, which was purified by rapid chromatography by elution with DCM:MeOH at a ratio of 99:1 to give compound 6-2 (0.5 g, 55.5% yield), which was a yellow solid.

[0373] 1 H NMR: 400MHz, CDCl3δ6.66(d,J=2.1Hz,1H),6.53–6.47(m,1H),4.65(s,1H),4.22(d,J=4.9Hz,2H),3.59(s,2H),2.23(s,3H),1.46(s,9H).

[0374] 4. Synthesis of compound 6-3

[0375] Compound 6-2 (100 mg, 369.34 μmol, 1.0 eq) was dissolved in THF (2 mL), and bis(trichloromethyl) carbonate (37.26 mg, 125.58 μmol, 0.34 eq) was added dropwise at 25 °C. The mixture was stirred at 25 °C for 3 hours. TLC showed that the reaction was complete. The resulting mixture was used directly in the next reaction step.

[0376] The newly prepared isocyanate was added to 3-[5-(aminomethyl)-1-oxoisoindolin-2-yl]piperidine-2,6-dione hydrochloride (114.40 mg, 369.34 μmol, 1.0 eq) and TEA (112.12 mg, 1.11 mmol, 3.0 eq) dissolved in DMF (2 mL). The mixture was stirred at 25 °C for 16 hours. LCMS showed that the reaction was complete. The reaction mixture was directly purified by preparative HPLC and lyophilized to give compound 6-3 (30 mg, yield 14.2%) as a white solid. LCMS: 514.30 [Mt-Bu+H] + .

[0377] 5. Synthesis of Compound 6

[0378] Compound 6-3 (20 mg, 0.0351 mmol, 1.0 eq) was dissolved in 1,4-dioxane (2 mL), and hydrochloric acid / dioxane (4 M, 1 mL) was added. The resulting mixture was stirred at 25 °C for 3 hours. TLC showed that the reaction was complete. The resulting solution was concentrated to give compound 6 (18 mg), which was used directly in the next step of the reaction.

[0379] 6. Synthesis of LPO3

[0380] To a DMF (1 mL) solution of compound 6 (32.15 mg, 52.13 μmol, 1.1 eq.), NMM (14.38 mg, 142.19 μmol, 3.0 eq.), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-morpholinium tetrafluoroborate (17.00 mg, 52.13 μmol, 1.1 eq.), and compound L2 (24 mg, 47.40 μmol, 1.0 eq.) were added. The resulting mixture was stirred at 25 °C for 4 hours. LC-MS showed that the reaction was complete. The reaction mixture was directly purified by preparative HPLC and lyophilized to give LP03 (24 mg, 47.3% yield) as a white solid with a purity of 99.17%. 1 H NMR (400MHz, DMSO-d6): δ10.98(s,1H),8.79(s,1H),8.63(t,J=6.5Hz,1H),8.3 3(t,J=5.7Hz,1H),8.24(t,J=5.9Hz,1H),8.14(d,J=7.9Hz,1H),8.07(t,J=5.7 Hz,1H),8.01(t,J=5.6Hz,1H),7.73–7.66(m,2H),7.50(s,1H),7.43(d,J=7.9H z,1H),7.27–7.15(m,6H),6.99(s,3H),6.76(t,J=5.7Hz,1H),5.15–5.06(m,1H ),4.63(d,J=6.6Hz,2H),4.51–4.38(m,4H),4.34–4.23(m,3H),3.93(s,2H),3. 80–3.55(m,7H),3.09–3.01(m,1H),2.96–2.84(m,1H),2.80(dd,J=13.8,9.7Hz ,1H),2.69–2.60(m,1H),2.45–2.30(m,2H),2.21(s,3H),2.10(t,J=7.4Hz,2H) ,2.03–1.95(m,1H),1.52–1.41(m,4H),1.22–1.13(m,2H).LC-MS:1068.6[M+H] + .

[0381] Example 9: Synthesis of LP04

[0382] Compound L1 (175.77 mg, 238.25 μmol, 1.2 eq.) was added to a DMF (2 mL) solution of compound 5 (110 mg, 198.54 μmol, 1 eq.) at 25 °C, and the mixture was stirred for 5 min. Then, HOBT (26.83 mg, 198.54 μmol, 1.0 eq.), pyridine (2 mL), and DIPEA (25.66 mg, 198.54 μmol, 1.0 eq.) were added to the mixture, and the resulting mixture was stirred at 25 °C for 16 h. LC-MS showed that the reaction was complete. The mixture was quenched with water (20 mL) and extracted with EA (3 × 20 mL). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by preparative HPLC (formic acid buffer) to give LP₀₄ (14.5 mg, 6.34% yield) as a grayish-white solid.

[0383] 1 H NMR (400MHz, DMSO-d6): δ10.97(s,1H),9.96(s,1H),8.85(s,1H),8.07(d,J=7.3Hz,1H),7.79(d,J=8.7Hz,1H),7.73–7.63(m,2H),7.51(s, 3H),7.44(d,J=7.8Hz,1H),7.06(s,1H),6.97(d,J=6.5Hz,3H),6.72(t,J=5.1Hz,1H),5.98(t,J=5.7Hz,1H),5.40(s,2H),5.13–5.05(m,1H ),4.47–4.30(m,6H),4.21–4.14(m,1H),3.83(d,J=9.7Hz,5H),3.05–2.84(m,3H),2.69–2.55(m,3H),2.44–2.31(m,1H),2.24–2.06(m,5H) ,2.03–1.90(m,2H),1.75–1.62(m,3H),1.58(d,J=9.2Hz,3H),1.53–1.31(m,7H),1.24–1.12(m,2H),0.88–0.77(m,6H).LC-MS:1152.7[M+H] + .

[0384] Example 10: Synthesis of Compound 7

[0385] 1. Synthesis of compound 7-2

[0386] o-Carboxybenzaldehyde (2 g, 13.3 mmol) was dissolved in methanol (20 mL), and a methanol solution of methylamine (30% wt) (2.5 g, 26.7 mmol) and sodium borohydride (253 mg, 6.658 mmol) were added. The mixture was stirred at room temperature for 2 hours. The reaction solution was filtered directly, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: methanol-dichloromethane, gradient: 0-50%) to give compound 7-2 (0.8 g, 36.4%), which was a colorless oil.

[0387] MS m / z(ESI): 166.1 [M+1].

[0388] 2. Synthesis of Compound 7-3

[0389] Compound 7-2 (800 mg, 4.846 mmol) was dissolved in 1,4-dioxane (20 mL), and water (20 mL), sodium hydroxide (776 mg, 19.4 mmol), and di-tert-butyl dicarbonate (2.1 g, 9.674 mmol) were added. The mixture was stirred at room temperature for 2 hours. The reaction solution was filtered directly, and the filtrate was concentrated under reduced pressure. The residue was purified by C18 silica gel column chromatography (mobile phase: acetonitrile-water (0.05% trifluoroacetic acid), gradient: 5%-75%) to give compound 7-3 (150 mg, 11.7%), which was a white solid.

[0390] MS m / z(ESI): 266.1 [M+1].

[0391] 3. Synthesis of compound 7-4

[0392] Compound 7-5 (35 mg, 0.056 mmol) was dissolved in N,N-dimethylformamide (1 mL), and N,N-diisopropylethylamine (21.8 mg, 0.169 mmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (21.4 mg, 0.056 mmol) and compound 7-3 (17.9 mg, 0.068 mmol) were added. The mixture was stirred at room temperature for 2 hours. The reaction solution was diluted with methanol (3 mL) and purified directly by C18 silica gel column chromatography (mobile phase: acetonitrile-water (0.05% formic acid), gradient: 5%-75%) to give compound 7-4 (18.6 mg, 38.0%), which was a white solid.

[0393] MS m / z (ESI): 869.4 [M+1].

[0394] 4. Synthesis of Compound 7

[0395] Compound 7-4 (13 mg, 0.015 mmol) was dissolved in dichloromethane (1.5 mL), and trifluoroacetic acid (0.5 mL) was added. The mixture was stirred at room temperature for 2 hours. The reaction solution was directly evaporated to dryness and then lyophilized to give compound 7 (15.8 mg), which was a white solid.

[0396] 1 H NMR (400MHz, MeOD) δ8.67(s,1H),8.52(s,1H),8.24(s,1H),7.96(dd,J=5.7,3.2Hz,1H),7.86–7.81(m,1H),7.78(d,J =8.9Hz,1H),7.74–7.65(m,4H),7.51(t,J=7.7Hz,1H),7.45(d,J=7.4Hz,1H),7.27(d,J=7.9Hz,1H),5.42(s,2H),5.1 9(dd,J=13.3,5.1Hz,1H),4.57(q,J=17.3Hz,2H),4.31(s,2H),4.10(s,4H),3.48(t,J=11.5Hz,1H),2.98–2.87(m,1H ),2.79(d,J=12.2Hz,4H),2.62–2.50(m,1H),2.36–2.15(m,5H),1.92(dd,J=24.4,11.8Hz,2H),1.85–1.71(m,2H).MS m / z(ESI):769.3[M+1].

[0397] Example 11: Synthesis of Compound 8

[0398] 1. Synthesis of compound 8-2

[0399] Compound 8-1 (100 mg, 0.16 mmol) was dissolved in N,N-dimethylformamide (3 mL), and triethylamine (48 mg, 0.48 mmol) and phenyl p-nitrochloroformate (38 mg, 0.19 mmol) were slowly added at 0 °C. The mixture was stirred at room temperature for 1 hour to give compound 8-2. This reaction was carried out directly in the next step without purification. MS m / z (ESI): 801.26 [M+H].

[0400] 2. Synthesis of compound 8-3

[0401] To a solution of compound 8-2 (100 mg, 0.13 mmol) in N,N-dimethylformamide (3 mL), triethylamine (40 mg, 0.39 mmol) and tert-butyl methyl (2-(methylamino)ethyl)carbamate (37 mg, 0.20 mmol) were slowly added at 0 °C, and the mixture was stirred at room temperature for 4 hours. The reaction mixture was quenched with water (50 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by C18 silica gel column chromatography (mobile phase: acetonitrile-water (0.05% trifluoroacetic acid), gradient: 5%-70%) to give compound 8-3 (40 mg, 36.2%) as a yellow-green solid.

[0402] MS m / z (ESI): 850.4 [M+1].

[0403] 3. Synthesis of Compound 8

[0404] Compound 8-3 (20 mg, 0.024 mmol) was dissolved in dichloromethane (4 mL), and trifluoroacetic acid (1 mL) was added. The mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and then lyophilized to give compound 8 (16.5 mg, 95.0%), which was a yellow-green solid.

[0405] 1 H NMR (400MHz, DMSO) δ8.48(d,J=5.0Hz,1H),8.01(d,J=9.2Hz,2H),7.73–7.60(m,2H),7.47(d,J=4.2Hz,1H),7.40– 7.29(m,2H),6.98(d,J=7.1Hz,1H),6.81–6.76(m,1H),6.72(d,J=8.5Hz,1H),4.99(dd,J=12.4,5.5Hz,1H),4.62( s,2H),4.04–3.86(m,4H),3.78(s,1H),3.63(t,J=5.5Hz,1H),3.41–3.23(m,2H),3.11(s,2H),2.98(s,1H),2.84– 2.53(m,6H),2.09(ddd,J=17.8,15.2,8.6Hz,5H),1.79(dd,J=23.3,12.5Hz,2H),1.63(dd,J=22.8,12.1Hz,2H).MS m / z(ESI):750.3[M+H].

[0406] Example 12: Synthesis of Compound 9

[0407] 1. Synthesis of compound 9-2

[0408] Compound 9-1 (85 mg, 0.13 mmol) was dissolved in N,N-dimethylformamide (2 mL), and acetoxyacetyl chloride (20 mg, 0.15 mmol) and triethylamine (40 mg, 0.39 mmol) were added. The mixture was stirred at room temperature for 2 hours. The reaction solution was quenched dropwise in ice water (20 mL), extracted with ethyl acetate (20 mL × 3), and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 9-2 (60 mg, crude product), which was a yellow solid. The product was used directly in the next reaction without purification.

[0409] MS m / z(ESI): 768.2 [M+1].

[0410] 2. Synthesis of Compound 9

[0411] Compound 9-2 (40 mg, 0.052 mmol) was dissolved in tetrahydrofuran (2 mL), and potassium carbonate (22 mg, 0.15 mmol) and methanol (2 mL) were added. The mixture was stirred at room temperature for 1 hour. The reaction solution was filtered, concentrated under reduced pressure, and the residue was purified by C18 silica gel column chromatography (mobile phase: acetonitrile-water (0.05% trifluoroacetic acid), gradient: 5%-45%) to give compound 9 (11.1 mg, 28.1%) as a yellow solid.

[0412] 1 H NMR (400MHz, DMSO) δ11.07(s,1H),10.16(s,1H),8.86(d,J=2.3Hz,1H),8.66(d,J=2.3Hz,1H),7.98(d,J=7.8Hz,1H),7.69( d,J=2.2Hz,1H),7.45(ddd,J=10.7,8.4,4.8Hz,2H),7.20(d,J=5.7Hz,2H),7.10–7.05(m,2H),6.96(dd,J=15.9,7.8Hz,3H) ,5.03(dd,J=12.9,5.3Hz,1H),4.48(d,J=6.1Hz,2H),4.04(s,2H),3.89–3.72(m,6H),3.08(ddd,J=11.8,8.5,3.4Hz,1H),2 .91–2.78(m,1H),2.18(d,J=12.1Hz,2H),2.05–1.95(m,3H),1.68(dt,J=12.0,10.2Hz,2H),1.47(dt,J=24.0,6.1Hz,2H).MS m / z(ESI):725.9[M+H].

[0413] Example 13: Synthesis of LP05

[0414] Compound L1 (40 mg, 0.054 mmol) was dissolved in pyridine (2 mL), and compound 8 (40.7 mg, 0.054 mmol) was added. The mixture was stirred for 18 hours. The reaction solution was diluted with methanol (5 mL) and purified by C18 silica gel column chromatography (mobile phase: acetonitrile-water (0.05% trifluoroacetic acid), gradient: 5%-60%) to obtain a crude product, which was a yellow oil. The crude product was purified by high performance liquid chromatography (column: Gemini 5u C18 150 x 21.2 mm, mobile phase: acetonitrile-water (0.1% trifluoroacetic acid), gradient: 30-70%) to obtain LP05 (10.7 mg, 14.9%), which was a white solid.

[0415] 1 H NMR(400MHz,MeOD)δ8.57(d,J=5.0Hz,1H),8.12(s,1H),7.64–7.58(m,1H),7.5 6(s,1H),7.45(t,J=7.8Hz,3H),7.34(s,2H),7.21(t,J=13.3Hz,2H),7.07(d,J= 7.1Hz,1H),6.82(d,J=8.5Hz,1H),6.74(s,2H),5.05(d,J=24.5Hz,3H),4.71(s, 2H),4.41(dd,J=16.0,6.6Hz,1H),4.27(s,1H),4.15–4.07(m,1H),3.95(s,1H), 3.87(d,J=22.6Hz,1H),3.58(s,3H),3.41(s,2H),3.19(dd,J=14.6,7.3Hz,4H), 3.04(d,J=8.0Hz,2H),2.99–2.94(m,4H),2.85(d,J=13.9Hz,2H),2.77–2.70(m, 2H),2.25(t,J=7.3Hz,3H),2.21(s,2H),1.87(d,J=11.6Hz,3H),1.71(d,J=7.3H z,3H),1.62–1.50(m,6H),1.30(d,J=7.3Hz,5H),0.95(dd,J=8.5,6.2Hz,6H).MS m / z(ESI): 1348.3 [M+H].

[0416] Example 14: Synthesis of LP06

[0417] Compound L2 ((S)-10-benzyl-23-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-6,9,12,15,18-pentoxo-3-oxo-5,8,11,14,17-pentazatriacid) (90 mg, 0.146 mmol) was dissolved in N,N-dimethylformamide (5 mL), and N,N-diisopropylethylamine (38 mg, 0.292 mmol), 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (83 mg, 0.219 mmol) and compound 9-1 (97 mg, 0.146 mmol) were added. The mixture was stirred at room temperature for 1 hour. The reaction solution was diluted with ethyl acetate (50 mL), washed with saturated saline (50 mL × 3), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by thin-layer chromatography with dichloromethane / methanol as the developing solvent to give LP06 (17.1 mg, 9.2%), which was a yellow solid.

[0418] 1 H NMR (400MHz, DMSO) δ11.10(s,1H),10.24(s,1H),8.85(d,J=2.1Hz,1H),8.77(t,J=6.4Hz,1H),8.66(d,J=2.1Hz,1H),8.36(d,J=5. 8Hz,1H),8.16(d,J=7.9Hz,1H),8.01(d,J=7.6Hz,3H),7.73(d,J=2.2Hz,1H),7.55–7.42(m,2H),7.29–7.09(m,7H),7.04–6.92(m, 4H),5.07(dd,J=12.8,5.4Hz,1H),4.71(d,J=6.6Hz,2H),4.52(d,J=6.0Hz,3H),4.14(s,2H),3.90–3.54(m,10H),3.09(d,J=11.6H z,3H),2.95–2.77(m,2H),2.71–2.54(m,2H),2.25–1.97(m,8H),1.71(dd,J=23.5,11.7Hz,2H),1.57–1.40(m,6H),1.26(s,2H).MS m / z(ESI):1266.4[M+1].

[0419] LP07 is the control molecule, and its structure is as follows: Prepared according to the method described in patent document WO2021198965.

[0420] Example 15: Synthesis of Compound 10

[0421] Synthesis of compound 10-1:

[0422] 1-(3-(aminomethyl)-5-chloro-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)methyl)urea (500 mg, 0.98 mmol), tetrahydro-4H-pyran-4-one (118.6 mg, 1.18 mmol), diethanolamine (638.0 mg, 4.93 mmol), and STAB (2092.7 mg, 9.87 mmol) were dissolved in DMF (10 mL) and stirred at 25 °C for 1 hour. The solvent was removed under reduced pressure, and the crude product was purified by flash evaporation with methanol (containing 0.1% NH3) at DCM = 0–10% to give a yellow solid product 10⁻¹ (300 mg, 52% yield). LC-MS (ESI, m / z): 553.9 [M+H] + .

[0423] Synthesis of compound 10-2:

[0424] To a 2 mL solution of compound 10-1 in DMF, 2-((tert-butyldimethylsilyl)oxy)ethyl(4-nitrophenyl)carbonate (40 mg, 0.11 mmol), triethanolamine (27.3 mg, 0.27 mmol), and HOBT (73.13 mg, 0.54 mmol) were added. The reaction was stirred at 25 °C for 1 hour, quenched with water, extracted with ethyl acetate, and the organic phase was concentrated under vacuum. The residue was purified by flash evaporation with methanol (DCM = 0–10%) to give a white solid product (40 mg, 55.65% yield). LC-MS (ESI, m / z): 756.0 [M+H] + .

[0425] Synthesis of compound 10:

[0426] To a 2 mL THF solution of compound 10-2 (40 mg, 0.07 mmol), TBAF (0.23 mL, 1 M THF solution) was added. The reaction was stirred at 25 °C for 1 hour, quenched with water, extracted with ethyl acetate, and the organic phase was concentrated under vacuum. The residue was purified by reversed-phase HPLC using an ACN-H2O (0.1% TFA) gradient to give a white solid product 10 (23.9 mg, 58.79% yield). 1H NMR(400MHz,DMSO)δ10.98(s,1H),8.86(s,1H),7.70–7.66(m,2H),7.51(s,1H),7.43(d,J=7.7Hz,1H), 6.88(s,1H),6.76(s,1H),5.11(dd,J=13.3,5.1Hz,1H),4.51–4.23(m,7H),4.10–3.95(m,3H),3.87–3. 82(m,2H),3.43–3.37(m,1H),3.32–3.36(m,2H),2.94–2.87(m,1H),2.71–2.52(m,2H),2.40–2.33(m,1 H),2.22(s,3H),2.02–1.97(m,1H),1.73–1.64(m,2H),1.62–1.53(m,2H).LC-MS(ESI,m / z):642.2[M+H] + .

[0427] Example 16: Synthesis of Compound 11

[0428] Synthesis of compound 11-1:

[0429] 2-((tert-butyldiphenylsilyl)oxy)acetic acid (68.1 mg, 0.21 mmol) was dissolved in DMF (2 mL), and DIEA (140.0 mg, 1.08 mmol) and HATU (123.54 mg, 0.32 mmol) were added. The mixture was stirred at 25 °C for 5 minutes, and then 1-(3-chloro-4-methyl-5-(((tetrahydro-2H-pyran-4-yl)amino)methyl)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)methyl)urea (60 mg, 0.10 mmol) was added. The reaction mixture was stirred at 25 °C for 1 hour. After the reaction was completed, the reaction was quenched with water and extracted with ethyl acetate. The organic phase was concentrated under vacuum, and the residue was purified by flash evaporation with methanol (DCM = 0–10%) to give a white solid product (50 mg, yield 51%). LC-MS (ESI, m / z): 850.3 [M+H] + .

[0430] Synthesis of compound 11:

[0431] To a THF solution of compound 11-1 (42 mg, 0.05 mmol), TBAF (0.14 mL, 1 M solution in THF) was added, and the reaction mixture was stirred at 25 °C for 1 hour. After the reaction was complete, the reaction mixture was concentrated under vacuum, and the residue was purified by reversed-phase HPLC using an ACN-H2O (0.1% TFA) gradient to give a white solid product 11 (17.9 mg, yield 57%). 1 H NMR (400MHz, DMSO) δ10.99(s,1H),8.90(s,1H),8.70(s,1H),7.70–7.64(m,2H),7.50(s,1H),7. 43(d,J=7.9Hz,1H),6.67–6.72(s,1H),6.76–6.72(m,1H),5.11(dd,J=13.3,5.0Hz,1H),4.66–4. 28(m,9H),3.91–3.82(m,3H),3.38–3.35(s,1H),2.95–2.87(m,1H),2.59(d,J=17.0Hz,1H),2.4 0–2.33(m,1H),2.24(s,3H),2.02–1.96(m,1H),1.69–1.51(m,4H).LC-MS(ESI,m / z):611.8[M+H] + .

[0432] Example 17: Synthesis of Compound 12

[0433] Synthesis of compound 12:

[0434] 1-[3-(aminomethyl)-5-chloro-4-methylphenyl]-3-{[2-(2,6-dioxopiperidin-3-yl)-1-oxo-3H-isoindol-5-yl]methyl}urea (50 mg, 0.10 mmol), 3-hydroxypropionic acid (9 mg, 0.10 mmol), EDCI (61 mg, 0.32 mmol), HOBT (43 mg, 0.32 mmol), and DIEA (137 mg, 1.0 mmol) were dissolved in DMF (1 mL) and stirred at 25 °C for 1 hour. The reaction solution was lyophilized, and the residue was purified by reverse-phase HPLC using an ACN-H2O (0.1% TFA) gradient to give a white solid product 12 (7.4 mg, 12.22% yield). 1H NMR (400MHz, DMSO) δ11.00(s,1H),8.82(s,1H),8.26–8.21(m,1H),7.71–7.66(m,2H),7.51(s,1H),7.44 (d,J=8.0Hz,1H),7.03(d,J=2.0Hz,1H),6.86–6.81(m,1H),5.11(dd,J=13.3,5.1Hz,1H),4.65–4.54(m, 1H),4.48–4.39(m,3H),4.34–4.19(m,3H),3.64(t,J=6.5Hz,2H),2.94–2.87(m,1H),2.66–2.57(m,1H), 2.41–2.35(m,1H),2.30(t,J=6.6Hz,2H),2.19(s,3H),2.02–1.97(m,1H).LC-MS(ESI,m / z):542.0[M+H] + .

[0435] Example 18: Synthesis of Compound 13

[0436] Synthesis of compound 13-1:

[0437] 1,1-Dimethoxypropane-2-one (500 mg, 4.23 mmol) and methyl magnesium bromide (5.0 mmol) were dissolved in 4 mL of diethyl ether and stirred at 0 °C for 1 hour under nitrogen protection. Then, trifluoroacetic acid (1930 mg, 16.9 mmol) and water (762 mg, 42.3 mmol) were added, and the reaction mixture was stirred at 30 °C for 1 hour. The reaction was monitored by thin-layer chromatography until completion. After the reaction was complete, the pH was adjusted to 7 and the mixture was used directly in subsequent steps.

[0438] Synthesis of compound 13:

[0439] To the reaction solution of compound 13-1, 1 mL of 1-[3-(aminomethyl)-5-chloro-4-methylphenyl]-3-{[2-(2,6-dioxopiridin-3-yl)-1-oxo-3H-isoindol-5-yl]methyl}urea (60 mg, 0.12 mmol) in DMF and STAB (81.2 mg, 0.38 mmol) were added. The reaction solution was stirred at 25 °C for 1 hour. After the reaction was completed, the reaction solution was concentrated under vacuum, and the residue was purified by Pre-HPLC (Gemini-C18 column: 150 × 21.2 mm, 5 μm; mobile phase: ACN-H2O (containing 0.1% FA); gradient: 20-25%) to give a white solid product 13 (10.9 mg, 14.96% yield). 1H NMR(400MHz,DMSO)δ10.99(s,1H),9.01(s,1H),8.59(s,2H),7.72–7.66(m,2H ),7.52–7.42(m,3H),7.16(t,J=5.9Hz,1H),5.24–5.02(m,2H),4.48–4.40(m, 3H),4.34–4.17(m,3H),2.94–2.90(m,2H),2.64–2.57(m,1H),2.44–2.35(m,1 H),2.31(s,3H),2.02–1.97(m,1H),1.19(s,6H).LC-MS(ESI,m / z):542.0[M+H] + .

[0440] Example 18: Synthesis of Compound 14

[0441] Synthesis of compound 14-1:

[0442] 1-[3-(aminomethyl)-5-chloro-4-methylphenyl]-3-{[2-(2,6-dioxopiridin-3-yl)-1-oxo-3H-isoindol-5-yl]methyl}urea (50 mg, 0.09 mmol), {2-[(tert-butyldimethylsilyl)oxy]ethyl}(4-nitrophenyl)carbonate (40 mg, 0.11 mmol), diethanolamine (38 mg, 0.3 mmol), and HOBT (40 mg, 0.3 mmol) were dissolved in DMF (1 mL) and stirred at 25 °C for 1 hour. After the reaction was complete, the reaction was quenched with water and extracted with ethyl acetate. The organic phase was concentrated under vacuum, and the residue was purified by flash evaporation (methanol:dichloromethane = 0–10%) to give a yellow solid product 14-1 (60 mg, 85.92% yield). LC-MS (ESI, m / z): 672.2 [M+H] + .

[0443] Synthesis of compound 14:

[0444] Compound 14-1 was dissolved in 2 mL of THF with TBAF (0.26 mL, 1 M). The reaction mixture was stirred at 25 °C for 1 hour. After the reaction was complete, the solvent was removed under vacuum, and the product was purified by Gemini-C18 pre-high performance liquid chromatography (150*21.2 mm, 5 μm mobile phase: ACN-H2O (0.1% TFA), gradient: 28-35) to give a white solid product 14 (18.5 mg, 35.27% yield). 1H NMR (400MHz, DMSO) δ10.99(s,1H),8.85(s,1H),7.74–7.63(m,3H),7.51(s,1H),7.44(d,J=8.0Hz,1H),7 .02–6.98(m,1H),6.76(t,J=5.9Hz,1H),5.11(dd,J=13.3,5.1Hz,1H),4.75(t,J=5.3Hz,1H),4.48–4.38 (m,3H),4.34–4.28(m,1H),4.18–4.11(m,2H),3.98(t,J=5.2Hz,2H),3.58–3.51(m,2H),2.96–2.87(m,1 H),2.63–2.55(m,1H),2.40–2.33(m,1H),2.20(s,3H),2.02–1.96(m,1H).LC-MS(ESI,m / z):557.8[M+H] + .

[0445] Example 19: Synthesis of Compound 15

[0446] 1-[3-(aminomethyl)-5-chloro-4-methylphenyl]-3-{[2-(2,6-dioxopiperidin-3-yl)-1-oxo-3H-isoindol-5-yl]methyl}urea solution (50 mg, 0.10 mmol), 3-hydroxycyclobutane-1-carboxylic acid (13.6 mg, 0.1 mmol), HATU (121 mg, 0.32 mmol), and DIEA (41 mg, 0.32 mmol) were dissolved in DMF (1 mL) and stirred at 25 °C for 1 hour. After the reaction was complete, 1 mL of water was added, and the product was purified by chromatography using a Gemini-C18 column (150 × 21.2 mm, 5 μm, mobile phase: ACN-H2O (0.1% FA), gradient: 28-28) to obtain a colored solid product 15 (8.6 mg, 15.70% yield). 1 H NMR (400MHz, DMSO) δ10.98(s,1H),8.75(s,1H),8.13(t,J=5.6Hz,1H),7.71–57.65(m,2H),7.51(s,1H),7.44(d,J=7.9Hz,1H),7.00( d,J=1.9Hz,1H),6.77(t,J=6.0Hz,1H),5.14–5.04(m,2H),4.50–4.38(m,3H),4.35–4.26(m,1H),4.22–4LC-MS(ESI,m / z):568.2[M+H] +..17(m,2H),3.97–3.88(m,1H),3.00–2.85(m,1H),2.65–2.56(m,1H), 2.47–2.34(m,2H),2.30–2.21(m,2H),2.18(s,3H),2.03–1.93(m,3H).

[0447] Example 20: Synthesis of Compound 16

[0448] 1-[3-(aminomethyl)-5-chloro-4-methylphenyl]-3-{[2-(2,6-dioxopiridin-3-yl)-1-oxo-3H-isoindol-5-yl]methyl}urea (60 mg, 0.12 mmol), (2R)-2-hydroxypropionic acid (21 mg, 0.24 mmol), HOBT (48 mg, 0.35 mmol), EDCI (68 mg, 0.35 mmol), and DIEA (92 mg, 0.71 mmol) were dissolved in DMF (1 mL) and stirred at 25 °C for 1 hour. After the reaction was complete, 1 mL of water was added, and the mixture was purified by passing it through a Gemini-C18 column (150 × 21.2 mm, 5 μm, mobile phase: ACN-H2O (0.1% FA), gradient: 28-28) to give a white solid product 16 (17.2 mg, 25.40% yield). 1 H NMR (400MHz, DMSO) δ10.99(s,1H),8.81(s,1H),8.11(t,J=5.9Hz,1H),7.72–7.66(m,2H),7.51(s,1 H),7.43(d,J=7.8Hz,1H),7.03–6.98(m,1H),6.87(t,J=5.9Hz,1H),5.10(dd,J=13.3,5.0Hz,1H),4. 47–4.38(m,3H),4.34–4.21(m,3H),4.03(q,J=6.7Hz,1H),2.95–2.86(m,1H),2.63–2.56(m,1H),2.4 3–2.33(m,1H),2.21(s,3H),2.03–1.97(m,1H),1.26(d,J=6.7Hz,3H).LC-MS(ESI,m / z):542.2[M+H] + .

[0449] Example 21: Synthesis of Compound 17

[0450] 1-[3-(aminomethyl)-5-chloro-4-methylphenyl]-3-{[2-(2,6-dioxopiridin-3-yl)-1-oxo-3H-isoindol-5-yl]methyl}urea solution (60 mg, 0.12 mmol), (2S)-2-hydroxypropionic acid (11 mg, 0.12 mmol), HOBT (52 mg, 0.38 mmol), EDCI (73 mg, 0.38 mmol), and DIEA (99 mg, 0.76 mmol) were added to DMF (1 mL) and stirred at 25 °C for 1 hour. 1 mL of water was added, and the mixture was purified by passing it through a Gemini-C18 column (150 × 21.2 mm, 5 μm, mobile phase: ACN-H2O (0.1% FA), gradient: 28-28) to give a white solid product 17 (12.8 mg, 17.54% yield). 1 H NMR (400MHz, DMSO) δ10.98(s,1H),8.75(s,1H),8.10(t,J=6.0Hz,1H),7.72–7.66(m,2H),7.51(s,1 H),7.43(d,J=7.9Hz,1H),6.99(d,J=1.9Hz,1H),6.83–6.76(m,1H),5.11(dd,J=13.3,5.1Hz,1H),4 .48–4.39(m,3H),4.33–4.21(m,3H),4.05–4.01(m,1H),2.95–2.85(m,1H),2.64–2.56(m,1H),2.43 –2.33(m,1H),2.21(s,3H),2.03–1.96(m,1H),1.26(d,J=6.8Hz,3H).LC-MS(ESI,m / z):542.2[M+H] + .

[0451] Example 22: Synthesis of Compound 18

[0452] 1-[3-(aminomethyl)-5-chloro-4-methylphenyl]-3-{[2-(2,6-dioxopiridin-3-yl)-1-oxo-3H-isoindol-5-yl]methyl}urea solution (50 mg, 0.10 mmol), 1-hydroxycyclopropane-1-carboxylic acid (21.7 mg, 0.21 mmol), diethanolamine (82 mg, 0.63 mmol), and HATU (121 mg, 0.32 mmol) were dissolved in DMF (2 mL) and stirred at 25 °C for 1 hour. After the reaction was complete, 1 mL of water was added, and the solution was purified by chromatography using a Gemini-C18 column (150 × 21.2 mm, 5 μm, mobile phase: ACN-H2O (0.1% FA), gradient: 28-28) to give a white solid product 18 (11 mg, yield 17.76%). 1 H NMR (400MHz, DMSO) δ10.99(s,1H),8.81(s,1H),8.28(t,J=6.2Hz,1H),7.74(d,J=2.1Hz,1H),7.69(d,J=7.8Hz ,1H),7.51(s,1H),7.44(d,J=8.0Hz,1H),6.97(d,J=2.0Hz,1H),6.85–6.80(m,1H),6.29(s,1H),5.13–5.08(m ,1H),4.48–4.40(m,3H),4.34–4.25(m,3H),2.94–2.87(m,1H),2.64–2.57(m,1H),2.41–2.35(m,1H),2.23(s, 3H),2.02–1.97(m,1H),1.04(dd,J=7.4,4.1Hz,2H),0.85(dd,J=7.4,4.1Hz,2H).LC-MS(ESI,m / z):554.2[M+H] + .

[0453] Example 23: Synthesis of LP08

[0454] 1-[3-(aminomethyl)-5-chloro-4-methylphenyl]-3-{[2-(2,6-dioxopiperidin-3-yl)-1-oxo-3H-isoindol-5-yl]methyl}urea (20 mg, 0.04 mmol), {[(2S)-2-[(2S)-2-[(2S)-2-[6-(2,5-dioxopyrrole-1-yl)hexamethylene]propionamide]propionamide]propionamide]methoxy}acetic acid (compound L3) (20.2 mg, 0.04 mmol), TEA (24 mg, 0.23 mmol), and DMTMMT (15.5 mg, 0.05 mmol) were dissolved in a mixed solvent of DMF (1.5 mL) and H2O (0.3 mL). The reaction solution was stirred at 25 °C for 1 hour. After the reaction was complete, 2 mL of water was added, and the product was purified by pre-HPLC using a Gemini-C18 (150 x 21.2 mm, 5 μm; mobile phase: ACN---H2O (0.1% TFA) gradient: 30-35) to obtain a white solid product LP08 (15.6 mg, 38% yield).

[0455] Example 24: Synthesis of LP09

[0456] Synthesis of compound LP09-1:

[0457] (2S,3R,4S,5S,6S)-2-(2-(2-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)acetamyl)-4-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate (500 mg, 0.68 mmol), bis(4-nitrophenyl) carbonate (828 mg, 2.72 mmol), and diethanolamine (880 mg, 6.8 mmol) were dissolved in tetrahydrofuran (6 mL). The reaction mixture was stirred at 25 °C for 16 hours. After the reaction was completed, the mixture was quenched with water and extracted with ethyl acetate. The organic phase was concentrated under reduced pressure, and the residue was purified by flash evaporation with ethyl acetate (PE = 0–60%) to give a white solid product (500 mg, 77% yield). LC-MS (ESI, m / z): 900.2 [M+H] + .

[0458] Synthesis of compound LP09-2:

[0459] Dissolve (2S,3R,4S,5S,6S)-2-(2-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)acetamido)-4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-trimethyltriacetate (100 mg, 0.06 mmol) in DMF (10 mL), and add 1-{3-chloro-4-methyl-5-[(oxetane-4-ylamino)methyl]phenyl}-3-{[2-(2,6-dioxopiperidin-3-yl)-1-oxo-3H-isoindol-5-yl]methyl}urea (37 mg, 0.06 mmol), diethanolamine (13 mg, 0.1 mmol), and HOBT (9 mg, 0.6 mmol). The reaction mixture was stirred at 25°C for 16 hours. After the reaction was complete, the reaction was quenched with water and extracted with ethyl acetate. The organic phase was concentrated under vacuum, and the residue was purified by pre-prepared thin-layer chromatography using methanol:dichloromethane = 5:95 to give a white solid product (50 mg, 34% yield). LC-MS (ESI, m / z): 1314.3 [M+H] + .

[0460] Synthesis of compound LP09-3:

[0461] Dissolve (2S,3R,4S,5S,6S)-2-(2-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)acetamyl)-4-((((3-chloro-5-(3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)methyl)ureido)-2-methylbenzyl)(tetrahydro-2H-pyran-4-yl)carbamoyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate (50 mg, 0.04 mmol) in acetonitrile (5 mL), and add sodium hydroxide solution (0.7 mL, 1 M NaOH aqueous solution). Stir the reaction solution at 0 °C for 10 minutes. After the reaction was complete, the product was acidified with formic acid and then purified by pre-HPLC (concentration gradient 15%–25%) using water-acetonitrile (containing formic acid) as the solvent to give a white solid product (30 mg, 83.33% yield). LC-MS (ESI, m / z): 952.0 [M+H] + .

[0462] Synthesis of compound LP09:

[0463] (2S,3S,4S,5R,6S)-6-[2-(2-aminoacetamido)-4-({[({3-chloro-5-[({[2-(2,6-dioxopiridine-3-yl)-1-oxo-3H-isoindol-5-yl]methyl}carbamoyl)amino]-2-methylphenyl}methyl)(oxetane-4-yl)carbamoyl]oxy}methyl)phenoxy]-3,4,5-trihydroxyoxetane-2-carboxylic acid (30 mg, 0.03 mmol), 2,5-dioxopirol-1-yl 3-(2,5-dioxopirol-1-yl)propionate (8.3 mg, 0.03 mmol) and DIEA (6 mg, 0.04 mmol) were dissolved in DMF (1 mL) and stirred at 25 °C for 1 hour. After the reaction was complete, 2 mL of water was added, and a white solid product LP09 (10.8 mg, 29% yield) was obtained by using Gemini-C18 (150 x 21.2 mm, 5 μm; mobile phase: ACN---H2O (0.1% TFA) gradient: 30-35).

[0464] Example 25: Synthesis of LP10

[0465] Synthesis of compound LP10-1:

[0466] 2,5-Dioxopyrrolidone-1-yl-(2S)-2-{[(9H-fluorene-9-ylmethoxy)carbonyl]amino}propionate (2 g, 4.9 mmol) was dissolved in 1,4-dioxane (20 mL), followed by the addition of (2S)-2-[(2S)-2-aminopropionamido]propionic acid (0.94 g, 5.88 mmol), NaHCO3 (4.55 g, 5.39 mmol), and H2O (2 mL). The reaction mixture was stirred at 25 °C for 16 hours. After the reaction was complete, the solvent was removed under reduced pressure, and the residue was purified by grinding with ethyl acetate (10 mL) and hydrochloric acid (10 mL, 1 M) to give a white solid product (1.5 g, 67.55% yield). LC-MS (ESI, m / z): 454.2 [M+H] + .

[0467] Synthesis of compound LP10-2:

[0468] (2S)-2-[(2S)-2-{[(9H-fluorene-9-ylmethoxy)carbonyl]amino}propionamide]propionamide]propionic acid (1.5 g, 3.3 mmol) was dissolved in DMF (20 mL), followed by the addition of (4-aminophenyl)methanol (0.81 g, 6.6 mmol), HATU (1.5 g, 6.93 mmol), and diethanolamine (1.28 g, 0.99 mmol). The reaction mixture was stirred at this temperature for 1 hour. After the reaction was complete, the reaction was quenched with 50 mL of water and then extracted with ethyl acetate (50 mL × 2). The combined organic phases were dried over Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with ethyl acetate / petroleum ether (ethyl acetate ratio increased from 50% to 100% within 20 minutes) to give a yellow solid product (0.8 g, yield 43%). LC-MS (ESI, m / z): [M+H] + 559.2.

[0469] Synthesis of compound LP10-3:

[0470] To a stirred THF solution (400 mg, 0.71 mmol) of 9H-fluorene-9-methyl N-[(1S)-1-{[(1S)-1-{[(1S)-1-{[4-(hydroxymethyl)phenyl]carbamoyl}ethyl]carbamoyl}ethyl]carbamoyl}ethyl]carbamoyl ester (400 mg, 0.71 mmol), 4-nitrophenyl chloroformate (288 mg, 1.43 mmol) and pyridine (113 mg, 1.43 mmol) were added sequentially. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction mixture was quenched with 50 mL of water and then extracted with ethyl acetate (50 mL × 2). The combined organic phases were dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with ethyl acetate / petroleum ether (ethyl acetate ratio increased from 50% to 100%), to give a yellow solid product (300 mg, yield 57.74%). LC-MS(ESI,m / z):[M+H] + 724.2.

[0471] Synthesis of compound LP10-4:

[0472] Under stirring, 1-{3-chloro-4-methyl-5-[(oxetane-4-ylamino)methyl]phenyl}propionamide]propionamide]propionamide]phenyl}methyl(4-nitrophenyl)carbonate (300 mg, 0.4 mmol) was added sequentially to a DMF (5 mL) solution of 1-{3-chloro-4-methyl-5-[(oxetane-4-ylamino)methyl]phenyl}-3-{[2-(2,6-dioxopiperidin-3-yl)-1-oxo-3H-isoindol-5-yl]methyl}urea (85 mg, 0.15 mmol), diethanolamine (29.47 mg, 0.228 mmol), and HOBT (20.54 mg, 0.15 mmol) at room temperature for 16 hours. After the reaction was complete, the reaction mixture was quenched with water (20 mL) and then extracted with ethyl acetate (20 mL × 2). The combined organic phases were dried over Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by pre-thin-layer chromatography (CH₂Cl₂ / MeOH = 15:1) to give a white solid product (80 mg, 15% yield). LC-MS (ESI, m / z): [M+H] + 1138.2.

[0473] Synthesis of compound LP10-5:

[0474] Under stirring, piperidine (1 mL) was added to a DMF (2 mL) solution of {4-[(2S)-2-[(2S)-2-{[(9H-fluoren-9-ylmethoxy)carbonyl]amino}propionamide]propionamide]propionamide]phenyl}methyl N-({3-chloro-5-[({[2-(2,6-dioxopiperidin-3-yl)-1-oxo-3H-isoindol-5-yl]methyl}amino)amino]-2-methylphenyl}methyl)-N-(oxetane-4-yl)carbamate (75 mg, 0.065 mmol)}methyl. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction mixture was concentrated under reduced pressure to give the crude product. The crude product was used directly in subsequent steps without further purification. LC-MS (ESI, m / z): [M+H]+916.2.

[0475] Synthesis of compound LP10:

[0476] In a 2 mL solution of 4-[(2S)-2-[(2S)-2-[(2S)-2-aminopropamido]propamido]propamido}phenyl}methyl N-({3-chloro-5-[({[2-(2,6-dioxopiperidin-3-yl)-1-oxo-3H-isoindol-5-yl]methyl}carbamoyl)amino]-2-methylphenyl}methyl)-N-(oxetane-4-yl)carbamate (40 mg, 0.043 mmol), 2,5-dioxopyrrolidone-1-yl-6-(2,5-dioxopyrrolidone-1-yl)hexanoate (26.48 mg, 0.086 mmol) and diethanolamine (16.64 mg, 0.129 mmol) were added sequentially, and the reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, 2 mL of water was added, and the product was purified by reversed-phase HPLC using a Gemini-C18 (150 x 21.2 mm, 5 μm; mobile phase: ACN---H2O (0.1% TFA) gradient: 30-35) to obtain a white solid product (10.3 mg, 21.39% yield). 1 H NMR (400MHz, DMSO) δ10.99(s,1H),9.75(s,1H),8.84(s,1H),8.16(d,J=6.4Hz,2H ),8.03(d,J=6.7Hz,1H),7.72–7.68(m,2H),7.66–7.52(m,2H),7.51(s,1H),7.44 (d,J=8.2Hz,1H),7.38–7.26(m,1H),7.12–7.02(m,1H),7.03–6.90(m,3H),6.72– 6.65(m,1H),5.15–4.95(m,3H),4.46–4.30(m,6H),4.27–4.18(m,2H),3.90–3.78( m,2H),3.35–3.26(m,4H),2.94–2.86(m,1H),2.66–2.54(m,1H),2.49–2.44(m,2H ),2.39(d,J=4.7Hz,1H),2.37–2.32(m,1H),2.19(s,3H),2.06(t,J=6.9Hz,2H),2 .02–1.96(m,1H),1.76–1.64(m,2H),1.62–1.55(m,2H),1.49–1.37(m,4H),1.29( d,J=6.9Hz,2H),1.23(d,J=7.0Hz,3H),1.20–1.09(m,5H).LC-MS(ESI,m / z):[M+H] + 1108.6.

[0477] Example 26: Synthesis of LP11

[0478] A solution of 2,5-dioxopyrrolidone-1-yl 1-[3-(2,5-dioxopyrrolidone-1-yl)propamido]-3,6,9,12,15,18,21,24-octaoxaheptadecane-27-ester (60 mg, 0.08 mmol), 1-[3-(aminomethyl)-5-chloro-4-methylphenyl]-3-{[2-(2,6-dioxopyridine-3-yl)-1-oxo-3H-isoindol-5-yl]methyl}urea (48.4 mg, 0.09 mmol) and diisoethanolamine (33 mg, 0.26 mmol) was dissolved in 2 mL of DMF and stirred at 25 °C for 1 hour. After the reaction was completed, 1 mL of water was added, and the product was pre-purified using a Gemini-C18 column (150×21.2 mm, 5 μm) (mobile phase: ACN-H2O (containing 0.1% FA), gradient: 30-33) to obtain a white solid product (16.7 mg, yield 17.47%). 1 H NMR (400MHz, DMSO) δ10.99(s,1H),8.83–8.78(m,1H),8.26(t,J=5.5Hz,1H),8.02(t,J=5.4Hz,1H),7.71–7.65(m,2H),7.51(s,1 H),7.44(d,J=7.9Hz,1H),7.05(d,J=1.7Hz,1H),7.00(s,2H),6.83–6.77(m,1H),5.11(dd,J=13.3,5.1Hz,1H),4.43(dd,J=15.8, 11.7Hz,3H),4.48–4.39(m,1H),4.21(d,J=5.5Hz,2H),3.64–3.57(m,4H),3.52–3.47(m,28H),3.36–3.33(m,2H),3.14(q,J=5.7 Hz,2H),2.95–2.87(m,1H),2.64–2.57(m,1H),2.41–2.30(m,5H),2.19(s,3H),2.03–1.97(m,1H).LC-MS(ESI,m / z):1044.2[M+H] + .

[0479] Example 27: Preparation of Antibody

[0480] In this example, the antibodies targeted Her2 and B7H3, respectively. The antibody targeting Her2 was Herceptin (Roche), and the antibody targeting B7H3 was Ifinatamab, the sequences of which are derived from patent CN103687945A. Signal peptides were added to the light and heavy chain sequences of the antibodies, and codon optimization was performed. The entire genome was then synthesized, and the target fragment was digested with enzymes and ligated into the pCDNA3.4 expression vector. Sequencing confirmed the sequence correctness. Subsequently, plasmid transformation was performed, and high-purity plasmids were extracted using an endotoxin-free large-scale extraction kit. Sequencing ensured the correctness of the plasmid sequence. The amino acid sequences of the antibodies are listed in Table 4. The plasmid expressing the antibodies was transfected into Expi293F cells and cultured in a cell culture shaker at 37°C, 120 rpm, and 8% CO2 for 5 days. The supernatant was collected, and the antibodies were purified using Protein A affinity chromatography. The antibody was characterized and analyzed using SDS-PAGE and SEC-HPLC methods. Figure 1 shows the results of SDS-PAGE analysis of ifinatamab antibody under both reduced and normal conditions, and Figure 2 shows the results of SEC analysis of ifinatamab antibody. The results show that ifinatamab antibody has good stability, and SEC analysis showed a purity of 99.06%.

[0481] Table 4: Antibody Sequence Information

[0482] Example 28: Preparation of ADC Samples

[0483] The antibody concentration is generally 5-10 mg / mL, and it is replaced in PBS buffer. 0.2 M EDTA stock solution is added to adjust the final EDTA concentration to 2 mM, and 10% H3PO4 is added to adjust the pH of the reaction system to approximately 6.0. The mixture is then gently mixed by pipetting. Reduction is performed using tris(2-carboxyethyl)phosphine hydrochloride (TCEP) (MCE) to break the disulfide bonds between the antibody chains. The amounts of TCEP and Linker-Payload vary when preparing ADC samples with different DAR values. Specific addition amounts can be found in Table 5. This correspondence is for reference only; conditions may be changed depending on the specific reaction type. The antibody and Linker-Payload small molecules used in ADC preparation are the corresponding products prepared in the aforementioned examples.

[0484] Table 5: Reference Table for TCEP and Linker-Payload Addition Amounts in ADC Sample Preparation with Different DAR Values

[0485] Taking the preparation of DAR4 ADC samples as an example, 3 times the molar equivalent of TCEP to the antibody was added to the reaction system, and the mixture was placed in a 37°C constant-temperature shaker at 120 rpm for 2 hours. After reduction, coupling was performed. The linker-payload molecule to be coupled was pre-dissolved in DMSO, aliquoted, and frozen at -80°C. The extracted molecule was temporarily stored on ice. For the preparation of DAR4 ADC samples, the linker-payload was added at 6.25 times the molar equivalent of the protein, and the DMSO in the reaction system was controlled to not exceed 10% of the total volume. The linker-payload molecule was slowly added to the reaction system, mixed thoroughly, sealed with a sealing film, and placed in a 25°C constant-temperature shaker at 120 rpm for 2 hours. After coupling, a certain amount of cysteine ​​was added, and the mixture was placed in a four-dimensional rotary mixer at 4°C and mixed thoroughly for 30 minutes to bind with the uncoupled molecule, thus terminating the reaction.

[0486] The amount of cysteine ​​added can be calculated using the following formula: V(cysteine) = 2*(n(additional drug equivalent) - n(theoretical coupling drug equivalent)) / c(cysteine).

[0487] After the reaction is terminated, select a desalting column (Thermo Fisher Scientific / Tiandi Renhe) or an ultrafiltration tube (Millipore) of appropriate specifications according to the volume of the reaction system for desalting. Replace the original buffer with PBS to remove DMSO, residual TCEP, EDTA, small molecule drugs and cysteine ​​from the system.

[0488] The concentration of the concentrated ultrafiltration sample was detected by Nanodrop, and the sample was characterized and identified by SEC, SEC-MS, HIC and other indicators. The remaining sample was frozen and stored at -80℃.

[0489] Taking the preparation of sample HER-LP03-2 as an example, the specific experimental operation is explained. The goal is to prepare the Herceptin-LP03-Dar8 sample. 1 mg of Herceptin antibody was replaced in PBS buffer, resulting in a concentration of approximately 5 mg / mL. 0.2 M EDTA stock solution was added to adjust the final EDTA concentration to 2 mM, and 10% H3PO4 was added to adjust the pH of the reaction system to approximately 6.0. The mixture was then gently mixed by pipetting. Six equivalents of TCEP were added to the reaction system for reduction. The system was placed in a 37°C constant-temperature shaker at 120 rpm for 2 hours to open the disulfide bonds between the antibody chains. After reduction, coupling was performed. The LP03 molecule to be coupled was pre-dissolved in DMSO, ensuring that the DMSO content in the reaction system did not exceed 10% of the total volume. 12.5 equivalents of LP03 were slowly added to the reaction system, mixed thoroughly, sealed with sealing film, and placed in a 25°C constant-temperature shaker at 120 rpm for 2 hours. After coupling, 9 equivalents of 20 mM cysteine ​​were added, and the mixture was thoroughly mixed at 4°C for 30 min in a four-dimensional rotary mixer to bind with the uncoupled small molecules, thus terminating the reaction. After the reaction was terminated, the sample was desalted using a 30 kDa ultrafiltration tube (Millipore), and the original buffer was replaced with PBS to remove DMSO, residual TCEP, EDTA, small molecule drugs, and cysteine ​​from the system. The concentration of the concentrated ultrafiltered sample was determined by Nanodrop, and samples were taken for characterization and identification using SEC, SEC-MS, and HIC assays. The remaining samples were stored at -80°C.

[0490] Example 29: Characterization and Identification of ADC Samples

[0491] 1. SEC testing methods

[0492] SEC (Sequencing Analysis) is generally used to determine the purity and polymer content of ADC (Advanced Dioxide-Acting Hydrocarbon) samples. The instrument used is an Agilent 1260 HPLC system; the analytical column is a TSK gel G3000SWxl; the mobile phase is 10 mM PBS, pH 7.2; the acquisition method is: flow rate set at 1 mL / min; acquisition time at 14 min; and the acquisition wavelength is 280 nm. For SEC detection, a suitable amount of the ADC sample to be tested is pipetted into the HPLC vial. The typical injection volume is 50 μg, but an extra 5-10 μL of sample should be taken to avoid insufficient sample leading to air bubbles affecting the analytical results. The sample purity and polymer content are then analyzed based on the HPLC chromatogram.

[0493] The ADC samples in Table 6 were subjected to SEC detection using the method described above. The results are shown in Figure 3-10 and Table 6. Among them, the Herceptin-LP03 and Ifinatamab-LP03 samples had extremely low polymer ratios and extremely high monomer ratios (purity), approximately 98%. However, the Herceptin-LP07 sample had a relatively high polymer ratio, and the higher the Dar value, the higher the polymer ratio.

[0494] Table 6: Purity of different ADC samples

[0495] 2. SEC-MS DAR value detection method

[0496] The DAR value of ADC samples can generally be detected using SEC-MS. The instrument used is a Waters Class I UPLC coupled with Synapt G2 SiQ-TOF; the analytical column is an ACQUITY UPLC Protein BEH SEC Column. 1.7 μm, 2.1 mm x 150 mm; mobile phase: 50 mM CH3COONH4, natural pH; acquisition method: flow rate set to 0.065 mL / min; acquisition time: 10 min; acquisition of ion current signal and UV280 absorbance. For SEC-MS detection, proteins containing glycans or ADC samples require desaccharification before delivery. Desaccharification system: dilute the protein sample to a concentration of 2 mg / mL with PBS, add 2 μL of PNGase F desaccharidase (catalog number: 20407ES01, hereinafter the same), total volume 20 μL; incubate in a 37℃ water bath for 2 h. If the sample does not contain glycans, desaccharification is not necessary; simply dilute the sample to a concentration of 2 mg / mL with PBS. Use a pipette to transfer an appropriate amount of the sample to the HPLC vial. The general injection volume is 2 μg; when taking samples, take an extra 5-10 μL to avoid insufficient sample leading to air bubbles affecting the analytical results. The composition of protein samples can be analyzed by convolution analysis of MS mass spectra or the DAR value of ADC samples can be calculated.

[0497] The following is a reference formula for calculating the DAR value:

[0498] Where: n is the number of coupled drugs, and In is the intensity of its corresponding peak.

[0499] The ADC samples in Table 7 were subjected to SEC-MS detection according to the above method. The results are shown in Figure 11-21 and Table 7. The HER-LP03, IFI-LP03, HER-LP07, HER-LP05, HER-LP06, and HER-LP04 samples have different DAR values.

[0500] Table 7: SEC-MS DAR values ​​of different ADC samples

[0501] 3. HIC DAR value detection method

[0502] The DAR value of ADC samples can generally be detected using HIC. The instrument used was an Agilent 1260 high-performance liquid chromatograph; the analytical column was an Advance Bio HIC 4.6×100mm (p / n 685975-908); mobile phase A was 50mM Na₂HPO₄, 25% IPA, pH 7.0, and mobile phase B was 2M (NH₄)₂SO₄, 50mM Na₂HPO₄, pH 7.0. The data acquisition method was as follows:

[0503] Table 8: Gradient elution degree

[0504] During HPLC analysis, use a pipette to transfer an appropriate amount of sample into the HPLC vial. The typical injection volume is 10 μL. If the injection volume is too large, flow-through may occur. When taking samples, take an additional 5-10 μL to avoid insufficient sample intake, which could introduce air bubbles into the syringe and affect the analytical results. Analyze the protein sample components based on the HPLC chromatogram or calculate the DAR value of the ADC.

[0505] The following is a reference formula for calculating the DAR value:

[0506] The ADC samples in Table 9 were subjected to HIC detection using the method described above. The results are shown in Figures 22-23 and Table 9. The HIC-DAR value of the HER-LP03-1 sample was 3.82, and the HIC-DAR value of the HER-LP04-1 sample was 3.77, which are basically consistent with the SEC-MS detection results mentioned above.

[0507] Table 9: HIC-DAR values ​​of ADC samples

[0508] The present invention also prepared the following ADC:

[0509] Table 10

[0510] ADC18 (HER-MMAE-DAR4.74), ADC23 (HER-GGFG-Dxd-Dar6.63), and ADC31 (HER-GGFG-Dxd-Dar7.06) were used as controls, and each has the following structure:

[0511] Where Bm is Herceptin, and n is 6.63 and 7.06 respectively;

[0512] Where Bm is Herceptin, n = 4.74.

[0513] In addition, for ease of comparison, ADC23 (HER-GGFG-Dxd-Dar6.63), ADC31 (HER-GGFG-Dxd-Dar7.06), and ADC18 (HER-MMAE-DAR4.74) will also be referred to as ADC-C01, ADC-C02, and ADC-C03 respectively in the following text.

[0514] Example 30: Detection of Cell Antiproliferative Activity

[0515] Log-phase NCI-N87, BT474, SK-OV-3, MDA-MB-231, MV-4-11, MDA-MB-468, and HCC1569 cells were collected, washed with PBS, and trypsin was added. The culture flask was gently shaken to ensure complete coverage of the cells. The flask was incubated at 37°C for 1-3 minutes or until the cells became completely rounded. Cell morphology changes were monitored under a microscope. Digestion was stopped by adding culture medium, and the cells were collected. A cell counter counts cells and checks cell viability. The cells were further diluted in the culture medium to adjust the cell suspension concentration to achieve a cell density of 10,000 cells / 100 μL of culture medium per well (NCI-N87 cells), 8,000 cells / 100 μL of culture medium per well (BT474 cells), 2,000 cells / 100 μL of culture medium per well (SK-OV-3 cells), 2,000 cells / 100 μL of culture medium per well (MDA-MB-231 cells), 2,000 cells / 100 μL of culture medium per well (MDA-MB-468 cells), 10,000 cells / 100 μL of culture medium per well (MV-4-11 cells), or 4,000 cells / 100 μL of culture medium per well (HCC1569 cells). Then, 100 μL / well was seeded into a 96-well plate. Incubate overnight at 37℃ and 5% CO2. Dilute the antibody, ADC sample, or small molecule compound with complete culture medium at 3-fold serial dilutions, creating 9 gradients. Adjust the initial concentration according to the sample. Add 100 μL / well to a 96-well plate, with 3 replicates per group. Add PBS to the peripheral wells. Set up zero wells (culture medium, CCK8) and control wells (cells, culture medium, CCK8). Incubate at 37℃ for 4-5 days. Remove the cell culture plate, add 20 μL of CCK8 solution to each well, and continue incubation for 1-4 hours (depending on the degree of color development). Measure the absorbance at 450 nm in each well using a microplate reader. Process the data and calculate the IC50 value of the sample using a four-parameter curve fitting method.

[0516] The antiproliferative activity of small molecule compounds in NCI-N87, BT474, MV-4-11, or MDA-MB-468 cells was investigated using the methods described above. The results are shown in Figures 24-29 and Tables 11-15. Compound 1 showed approximately two orders of magnitude higher activity than SMol006 in the antiproliferative assay of NCI-N87 or BT474 cells, while compound 4 showed 3-10 times stronger activity than SMol006. In BT474 cells, compound 4 exhibited slightly stronger cytotoxic activity than Dxd, but due to the high sensitivity of NCI-N87 cells to Dxd, the cytotoxic activity of compound 4 in NCI-N87 cells was significantly lower than that of Dxd. In HCC1569 cells, compounds 4 and LP04 showed 6-16 times stronger activity than SMol006. Compared with compounds 4 and LP04, compounds 10, 11, 12, 13, 14, 15, 16, 17, and 18 also showed significant cytotoxic activity in MV-4-11 or MDA-MB-468 cells, and their IC50 values ​​are shown in Tables 13 and 14.

[0517] Table 11: Experimental results of the anti-proliferative activity of small molecule compounds in NCI-N87 cells

[0518] Table 12: Experimental results of the anti-proliferative activity of small molecule compounds in BT474 cells

[0519] Table 13: Experimental results of the anti-proliferative activity of small molecule compounds in MV-4-11 cells

[0520] Table 14: Experimental results of the antiproliferative activity of small molecule compounds in MDA-MB-468 cells

[0521] Table 15: Experimental results of the antiproliferative activity of small molecule compounds in HCC1569 cells

[0522] The antiproliferative activity of Herceptin antibody and ADC samples with different DAR values ​​in N87, BT474, SK-OV-3, MDA-MB-231, or HCC1569 cells was tested using the method described above. The results are shown in Figures 30-37 and Tables 16-23. In the N87 cell antiproliferative assay, HER-LP03 and HER-LP04 samples showed higher antiproliferative activity than HER-GGFG-Dxd or HER-LP07 samples. In the BT474 cell antiproliferative assay, HER-LP03 and HER-LP04 samples showed similar or higher antiproliferative activity than HER-LP07 samples. In the BT474 cell antiproliferative assay, HER-LP03 samples showed higher antiproliferative activity than HER-GGFG-Dxd samples. In the SK-OV-3 cell antiproliferative assay, HER-LP03 and HER-LP04 samples showed higher antiproliferative activity than HER-GGFG-Dxd samples. HER-LP03 and HER-LP04 samples showed higher anti-proliferative activity than HER-GGFG-Dxd and HER-MMAE samples in HCC1569 cell anti-proliferation assays. HER-LP03 and HER-LP04 samples showed higher anti-proliferative activity than HER-LP07 samples in HCC1569 cell anti-proliferation assays. ADC13 showed similar anti-proliferative activity to ADC12 (HER-LP03-Dar3.6) in N87 cell anti-proliferation assays, while ADC16 and ADC17 showed similar anti-proliferative activity to ADC15 in N87 cell anti-proliferation assays. MDA-MB-231 cells are triple-negative breast cancer cells; all ADC samples targeting Her2 did not show cytotoxic activity in anti-proliferation assays, indicating that the activity of ADCs is Her2-target mediated and targeted.

[0523] Table 16: Experimental results of the anti-proliferative activity of ADC samples in N87 cells

[0524] Table 17: Experimental results of the anti-proliferative activity of ADC samples in N87 cells

[0525] Table 18: Experimental results of the anti-proliferative activity of ADC samples in N87 cells

[0526] Table 19: Experimental results of the anti-proliferative activity of ADC samples in BT474 cells

[0527] Table 20: Experimental results of the anti-proliferative activity of ADC samples in BT474 cells

[0528] Table 21: Experimental results of the anti-proliferative activity of SK-OV-3 cells by ADC samples

[0529] Table 22: Experimental results of anti-proliferative activity of ADC samples in HCC1569 cells

[0530] Table 23: Experimental results of antiproliferative activity of ADC and small molecule samples in MDA-MB-231 cells

[0531] Note: NA indicates not applicable, ND indicates no detection.

[0532] Example 31: In vitro Bystander activity assessment of ADC samples

[0533] Her2-positive BT474 cells (Her2+) and Her2-negative MDA-MB-468 cells (Her2-) in the logarithmic growth phase were washed with PBS, and trypsin was added. The culture flasks were gently shaken to ensure complete coverage of the cells with trypsin. The flasks were incubated at 37°C for 1-3 minutes or until the cells were completely rounded. Cell morphology changes were monitored under a microscope. Digestion was stopped by adding culture medium, and the cells were collected. Cell counters count cells and check cell viability, using CellTrace. TM Violet Cell Proliferation Kit and CellTrace TMThe Far Red Cell Proliferation Kit was used to stain BT474 and MDA-MB-468 cells. After staining, the cell suspension concentration was adjusted with complete culture medium, and the cells were seeded into 4-well plates at different cell volumes.

[0534] A:Her2+:Her2- = 5:1, which means 8.33E5 BT474 cells + 1.67E5 MDA-MB-468 cells per well per 2 mL;

[0535] B:Her2+:Her2- = 2:1, which means 6.67E5 BT474 cells + 3.33E5 MDA-MB-468 cells per well per 2 mL;

[0536] C:Her2+:Her2- = 1:0, which means 1E6 BT474 cells per well per 2mL;

[0537] D:Her2+:Her2- = 0:1, which means 1E6 MDA-MB-468 cells per well per 2mL;

[0538] Incubate overnight at 37°C with 5% CO2. Dilute the ADC sample to 60 nM with complete culture medium, adding 2 mL / well to each well of a 4-well plate to a final concentration of 30 nM. Include blank cells (left and right control groups) in wells without ADC sample, with one replicate per group. Incubate at 37°C for 4-5 days. Collect the supernatant. Wash the remaining adherent cells with PBS, add trypsin, and gently shake the plate to ensure complete coverage of the cells. Incubate the plate at 37°C for 1-3 minutes or until the cells are completely rounded. Monitor cell morphology changes under a microscope. Add culture medium to stop digestion, centrifuge at 300g for 5 minutes, discard the supernatant, resuspend the cells in an appropriate amount of PBS, and analyze cell count and viability using a cell counter. Analyze the Her2+ / Her2- cell ratio using flow cytometry. As shown in Figures 38A-D, when the ratio of effector cells to target cells was 5:1 and 2:1, both ADC02 and ADC08 exhibited high Bystander activity, with ADC08 showing approximately 3 times higher Bystander activity than ADC05.

[0539] Example 32: In vivo activity assessment of ADC samples

[0540] The in vivo activity of the ADC samples was evaluated in N87 and SK-OV-3 mouse models.

[0541] 1. Collect N87 cells in the logarithmic growth phase, ensuring the cells are free of endotoxin contamination. After cell counting, add Matrigel gel at a 4:1 ratio to adjust the cell concentration to 1.0 × 10⁶ cells / year. 8Cells / mL, mix well and aliquot into 1.5mL EP tubes. Four-week-old female Babl / c nude mice (Vitol) were inoculated with cells one week after purchase and acclimatization. Inoculation was performed under the right back axilla of each mouse, with 0.1mL injected per mouse, equivalent to 1.0 × 10⁶ cells per mouse. 7 / mouse. Approximately 5 days after cell inoculation, the growth of the mouse tumor was observed daily. The longest and shortest sides of the tumor exposed on the outer side of the skin were measured using calipers, and the mouse's weight was recorded. Tumor volume was calculated as 0.5 * length * width * width(high). When the tumor grew to approximately 100 mm... 3 Mice were randomly divided into groups to ensure a consistent mean tumor volume across groups, with no significant differences between groups. Eight mice were assigned to each group, and as shown in Figure 39, different ADC samples or control samples were administered via tail vein at doses of 5 mg / kg or 10 mg / kg on Day 0 and Day 7, respectively. Tumor volume and body weight were measured twice weekly, and tumor volume growth curves were plotted using GraphPad. The statistical analysis was conducted to determine if there were significant differences in tumor volume between groups, and the body weight change of each mouse after each administration was also recorded.

[0542] The efficacy evaluation results of the N87 mouse model are shown in Figures 39 and 40. Figure 39 shows the tumor growth curves of different treatment groups, with arrows representing drug administration time points. Compared with the control group, all treatment groups showed inhibitory effects on tumor growth in N87 model mice, and the differences were statistically significant. Among them, ADC samples ADC12, ADC22, and ADC15 significantly inhibited tumor growth, with effects significantly superior to ADC21 and Herceptin. ADC12 showed a dose-dependent effect in the N87 mouse model, with 10 mg / kg showing a slightly better tumor inhibition rate than 5 mg / kg. Figure 40 shows the changes in body weight of tumor-bearing mice in different treatment groups. No significant decrease in mouse body weight was observed after drug administration in any group, indicating that the ADC drug of this invention has good safety.

[0543] 2. Collect SK-OV-3 cells in the logarithmic growth phase, ensuring the cells are free of endotoxin contamination. After cell counting, adjust the cell concentration to 4.0 × 10⁻⁶. 7 Cells / mL, added 1:1 to Matrigel gel, mixed well and dispensed into 1.5mL EP tubes. Four-week-old female Babl / c nude mice (Vitol) were inoculated with cells one week after purchase and acclimatization. Inoculation was performed under the right back axilla of each mouse, with 0.1mL injected per mouse, equivalent to 2.0 × 10⁶ cells per mouse. 6 / mouse. Approximately 13 days after cell inoculation, the growth of the mouse tumor was observed daily. The longest and shortest sides of the tumor exposed on the outer side of the skin were measured using calipers, and the mouse's weight was recorded. Tumor volume was calculated as 0.5 * length * width * width(high). When the tumor grew to approximately 100 mm...3 Mice were randomly divided into groups to ensure a consistent mean tumor volume across groups, with no significant differences between groups. Eight mice were assigned to each group, and as shown in Figure 41, different ADC samples or control samples were administered via tail vein at a dose of 10 mg / kg on Day 0 and Day 7, respectively. Tumor volume and body weight were measured twice weekly, and tumor volume growth curves were plotted using GraphPad. The statistical analysis was conducted to determine if there were significant differences in tumor volume between groups, and the body weight change of each mouse after each administration was also recorded.

[0544] The efficacy evaluation results of the SK-OV-3 mouse model are shown in Figures 41 and 42. Figure 41 shows the tumor growth curves of different treatment groups, with arrows representing drug administration time points. Compared with the control group, all treatment groups showed inhibitory effects on tumor growth in SK-OV-3 model mice, and the differences were statistically significant. Among them, ADC12, ADC15, and ADC22 samples significantly inhibited tumor growth, with effects significantly better than ADC21 and Herceptin. ADC22 showed a slightly better inhibitory effect on tumor growth in SK-OV-3 model mice than ADC12. Figure 42 shows the changes in body weight of tumor-bearing mice in different treatment groups. The body weight loss of mice in each ADC sample group remained within 5% after drug administration, indicating that the ADC drugs of this invention have good safety.

[0545] Example 33: Pharmacokinetics of ADC Samples in ICR Mice

[0546] ICR mice were randomly divided into experimental groups (n = 3 × 3) and administered the test sample via a single intravenous injection at a dose of 5 MPa. Whole blood samples were collected from mice at 0.0833 h, 2 h, 7 h, 24 h, 48 h, 72 h, 96 h, 168 h, and 336 h post-administration. Plasma was obtained after centrifugation. LC-MS / MS was used to detect free payload and intact ADC, and ELISA was used to detect total antibody in the samples.

[0547] The results are shown in Table 24. ADC21 (HER-LP07-Dar3.77) had a plasma half-life (T0.05) in ICR mice. 1 / 2 The time to total antibody levels in ICR mice was 25.1 h. 1 / 2 The free payload was detectable after 227 hours; ADC19 (HER-LP03-Dar3.60) was detected in the T cells of ICR mice. 1 / 2 The total antibody was detected in T cells of ICR mice at a time interval of 24.6 hours. 1 / 2 239h, no free payload detected; ADC22 (HER-LP03-Dar7.63) in ICR mice T 1 / 2The total antibody was detected in T cells of ICR mice at a time rate of 23.1 h. 1 / 2 After 185 hours, no free payload was detected; ADC15 (HER-LP04-Dar3.73) was detected in the T cells of ICR mice. 1 / 2 The total antibody was detected in T cells of ICR mice at a time interval of 29.3 hours. 1 / 2 The time was 135h, and no free payload was detected.

[0548] Table 24: Pharmacokinetic analysis of ADC samples in ICR mice

[0549] NA: Not applicable; Cmax: Peak concentration; Tmax: Time to peak concentration; AUC0-last: Area under the plasma concentration-time curve from time 0 to the final quantifiable time point; AUC0-inf: Area under the plasma concentration-time curve from time 0 to indefinite time after administration; Vss: Steady-state volume of distribution; CL: Clearance; MRT0-last: Mean residence time from time 0 to the final quantifiable time point; MRT0-inf: Mean residence time from time 0 to indefinite time after administration; t1 / 2: Half-life.

[0550] Example 34: Detection of the metabolic stability of ADC in human and CD1 mouse plasma

[0551] The plasma metabolic stability of ADCs was assessed using human plasma and CD-1 mouse plasma. ADC incubation group: ADC was added to human or CD-1 mouse plasma to a final concentration of 100 μg / ml. After thorough mixing, the mixture was incubated at 37°C for 4, 8, 24, 48, 96, 168, and 336 hours, respectively. Payload incubation group: Payload compound was added to human or CD-1 mouse plasma to a final concentration of 1 μM. After thorough mixing, the mixture was incubated at 37°C for 0, 5, 15, 30, 45, 60, and 120 minutes, respectively. The reaction was terminated immediately after each set time point, and the samples were frozen. Total antibodies in the samples were detected using ELISA. ADC and free payload in the samples were detected using LC-MS / MS.

[0552] The results are shown in Table 25. The plasma half-life (T0) of ADC22 (HER-LP03-Dar7.63) in human plasma is shown in Table 25. 1 / 2 The time to T in CD1 mouse plasma was 9.18 h. 1 / 2 The time to T in human plasma was 13.6 h; Compound 4's T in human plasma 1 / 2 The time to T in CD1 mouse plasma was 6.77 h. 1 / 2The time to T in total antibody in human plasma was 8.43 h. 1 / 2 For 462 hours, T in CD-1 mouse plasma 1 / 2 It takes 756 hours.

[0553] Table 25: Results of metabolic stability of ADC and payload in human and CD1 mouse plasma

[0554] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

[0555] The foregoing embodiments and methods described in this invention may vary based on the capabilities, experience, and preferences of those skilled in the art.

[0556] Listing the steps of the method in a certain order in this invention does not constitute any restriction on the order of the method steps.

Claims

1. A compound or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, or deuterated thereof, characterized in that, The compound has the following structure: in, U is selected from: C0-C 10 Alkyl, C3-C 10 cycloalkyl, C6-C 10 Aryl, 4-10 membered heterocyclic groups; V is selected from: -O-, -S-, B is The C ring is a divalent group attached to the A ring; it either does not exist or is selected from: C3-C 10 Cycloalkylene, C6-C 10 arylene, 4-10 membered heterocyclic arylene; X, Y, and Z are independently selected from: single bonds, C1-C 10 Alkylene, -(C0-C6 alkylene)-O-, -(C0-C6 alkylene)-S-, -(C0-C6 alkylene)-C(O)-, -(C0-C6 alkylene)-C(S)-, -(C0-C6 alkylene)-N(R) B1 )-、-(C0-C6 alkylene)-CON(R B1 )-、-(C0-C6 alkylene)-N(R B1 CO-, -(C0-C6 alkylene)-SO2-, -(C0-C6 alkylene)-SO-, -(C0-C6 alkylene)-(C3-C 10 Cycloalkylene)-, -(C0-C6 alkylene)-(C6-C 10 arylene)-, -(C0-C6 alkylene)-(4-10 heterocyclic)-; wherein, the C0-C 10 Alkylene, C3-C 10 Cycloalkylene, C6-C 10 The hydrogen atoms in the arylene and 4-10 membered heterocyclic groups may optionally be substituted with one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, -(C0-C6 alkylene)-(C3-C 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, C1-C 10 Halogenated alkoxy groups, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)CON(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -O(C) 0-10 alkyl), -S(C 0-10 Alkyl), -SO(C) 0-10 alkyl), -SO2(C 0-10 alkyl), -SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0- 10 alkyl); Ring A is selected from: C3-C 10 Cycloalkylene, C6-C 10 arylene, 4-10 membered heterocyclic; of which, C3-C 10 Cycloalkylene, C6-C 10 The hydrogen atoms in the arylene and 4-10 membered heterocyclic groups may optionally be substituted with one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, -(C0-C6 alkylene)-(C3-C 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, C1-C 10 Haloalkoxy, -(C0-C6 alkylene)-N(C 0-10 Alkyl)(C 0-10 Alkyl), -(C0-C6 alkylene)-N(C 0-10 Alkyl)CO(C 0-10 Alkyl), -(C0-C6 alkylene)-N(C 0-10 Alkyl)CON(C 0-10 Alkyl), -(C0-C6 alkylene)-N(C 0-10 Alkyl)SO2(C 0-10 Alkyl), -(C0-C6 alkylene)-O(C 0-10 Alkyl), -(C0-C6 alkylene)-S(C 0-10 Alkyl), -(C0-C6 alkylene)-SO(C 0-10 Alkyl), -(C0-C6 alkylene)-SO2(C 0-10 Alkyl), -(C0-C6 alkylene)-SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -(C0-C6 alkylene)-COO(C 0-10 Alkyl), -(C0-C6 alkylene)-OCO(C 0-10 Alkyl), -(C0-C6 alkylene)-CON(C 0-10 Alkyl)(C 0-10 Alkyl), -(C0-C6 alkylene)-CO(C 0-10 alkyl); D is L1, L2, and L3 are independently selected from: single bonds, C1-C 10 Alkylene, -O-(C0-C6 alkylene)-, -S-(C0-C6 alkylene)-, -N(R L0 )-(C0-C6 alkylene)-, -N(R L0 )C(O)-(C0-C6 alkylene)-、-N(R L0 )C(O)O-(C0-C6 alkylene)-、-N(R L0 )C(O)N(R L0 -(C0-C6 alkylene)-, -C(O)-(C0-C6 alkylene)-, -C(S)-(C0-C6 alkylene)-, -C(O)O-(C0-C6 alkylene)-, -OC(O)-(C0-C6 alkylene)-, -(C0-C6 alkylene)-OC(O)-, -CON(R L0 -(C0-C6 alkylene)-, -SO2-(C0-C6 alkylene)-, -SO-(C0-C6 alkylene)-, C3-C 10 Cycloalkylene, C6-C 10 arylene, 4-10 membered heterocyclic alkylene; wherein, the CO-C 10 Alkylene, C3-C 10 Cycloalkylene, C6-C 10 The hydrogen atoms in the arylene and 4-10 membered heterocyclic groups may optionally be substituted with one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, -(C0-C6 alkylene)-(C3-C 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, C1-C 10 Halogenated alkoxy groups, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)CON(C 0-10 alkyl), -N(C) 0- 10 Alkyl)SO2(C 0-10 Alkyl), -O(C) 0-10 alkyl), -S(C 0-10 Alkyl), -SO(C) 0-10 alkyl), -SO2(C 0-10 alkyl), -SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl); L represents the connector precursor; R V1 R V2 R B1 R L0 Independently selected from: H, deuterium, halogen, cyano, nitro, azide, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, -(C0-C6 alkylene)-(C3-C 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, C1-C 10 Halogenated alkoxy groups, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)CON(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -O(C) 0-10 alkyl), -S(C 0-10 Alkyl), -SO(C) 0-10 alkyl), -SO2(C 0-10 alkyl), -SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl).

2. The compound according to claim 1, characterized in that, U is selected from: C0-C6 alkyl, C3-C6 cycloalkyl, C6-C 10 Aryl, Preferably, W is selected from: -O-, -S-, Preferably, V is Preferably, X is selected from: C1-C 10 Alkylene, -(C0-C6 alkylene)-O-, -(C0-C6 alkylene)-S-, -(C0-C6 alkylene)-C(O)-, -(C0-C6 alkylene)-C(S)-, -(C0-C6 alkylene)-N(R) X1 )-、-(C0-C6 alkylene)-CON(R X1 )-、-(C0-C6 alkylene)-N(R X1 CO-, -(C0-C6 alkylene)-SO2-, -(C0-C6 alkylene)-SO-; Preferably, Y is selected from: -(C0-C6 alkylene)-O-, -(C0-C6 alkylene)-S-, -(C0-C6 alkylene)-C(O)-, -(C0-C6 alkylene)-C(S)-, -(C0-C6 alkylene)-N(R) Y1 )-、-(C0-C6 alkylene)-CON(R Y1 )-、-(C0-C6 alkylene)-N(R Y1 CO-, -(C0-C6 alkylene)-SO2-, -(C0-C6 alkylene)-SO-, -(C0-C6 alkylene)-(C3-C 10 Cycloalkylene)-, -(C0-C6 alkylene)-(C6-C 10 arylene)-, -(C0-C6 alkylene)-(4-10 heterocyclic)-; wherein, the C0-C6 alkylene, C3-C 10 Cycloalkylene, C6-C 10 The hydrogen atoms in the arylene and 4-10 membered heterocyclic groups may optionally be substituted with one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl); Preferably, Z is selected from: -(C0-C6 alkylene)-O-, -(C0-C6 alkylene)-S-, -(C0-C6 alkylene)-C(O)-, -(C0-C6 alkylene)-C(S)-, -(C0-C6 alkylene)-N(R) Z1 )-、-(C0-C6 alkylene)-CON(R Z1 )-、-(C0-C6 alkylene)-N(R Z1 CO-, -(C0-C6 alkylene)-SO2-, -(C0-C6 alkylene)-SO-; Preferably, R W1 R W2 R X1 R Y1 R Z1 Independently selected from: H, deuterium, halogen, cyano, nitro, azide, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, -(C0-C6 alkylene)-(C3-C 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, C1-C 10 Halogenated alkoxy groups, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)CON(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -O(C) 0-10 alkyl), -S(C 0- 10 Alkyl), -SO(C) 0-10 alkyl), -SO2(C 0-10 alkyl), -SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0- 10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl); Preferably, R V1 R V2 R W1 R W2 R X1 R Y1 R Z1 Independently selected from: H, deuterium, halogen, cyano, nitro, azide, C1-C 10 Alkyl, C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl); Preferably, ring A is selected from: Preferably, R L4 R L5 Independently selected from: deuterium, halogen, cyano, nitro, azide, C1-C 10 Alkyl, -(C0-C6 alkylene)-(C3-C6 alkylene) 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), -(C0-C6 alkylene)-N(C 0-10 Alkyl)(C 0-10 Alkyl), -(C0-C6 alkylene)-N(C 0-10 Alkyl)CO(C 0-10 Alkyl), -(C0-C6 alkylene)-O(C 0-10 Alkyl), -(C0-C6 alkylene)-COO(C 0-10 Alkyl), -(C0-C6 alkylene)-OCO(C 0-10 Alkyl), -(C0-C6 alkylene)-CON(C 0-10 Alkyl)(C 0- 10 Alkyl), -(C0-C6 alkylene)-CO(C 0-10 alkyl).

3. The compound according to claim 2, characterized in that, U is selected from: H, Preferably, ring C is absent or selected from: X is selected from: Preferably, Y is selected from: -O-, Z is selected from: -O-, Preferably, B is selected from: Preferably, ring A is selected from:

4. The compound according to claim 1, characterized in that, The compound shown in Formula I contains at least one deuterium atom; Preferably, one or more H atoms bonded to the carbon atom in D can be replaced by deuterium; Preferably, one or more H atoms bonded to the carbon atoms in L1, L2, and L3 can be replaced by deuterium; L1 is selected from: single bond, -O-(C0-C6 alkylene)-, -S-(C0-C6 alkylene)-, -N(R L1 )-(C0-C6 alkylene)-, -N(R L1 )C(O)-(C0-C6 alkylene)-、-N(R L1 )C(O)O-(C0-C6 alkylene)-、-N(R L1 )C(O)N(R L1 -(C0-C6 alkylene)-, -C(O)-(C0-C6 alkylene)-, -C(O)O-(C0-C6 alkylene)-, -CON(R L1 )-(C0-C6 alkylene)-; wherein, R L1 Selected from: H, deuterium, halogen, cyano, nitro, azide, C1-C 10 Alkyl, C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl); Preferably, L1 is selected from: single bond, -O-, Preferably, L2 is selected from: single bonds, C1-C 10 Alkylene, -O-(C0-C6 alkylene)-, -S-(C0-C6 alkylene)-, -N(R L2 )-(C0-C6 alkylene)-, -N(R L2 -C(O)-(C0-C6 alkylene)-, -C(O)-(C0-C6 alkylene)-, -CON(R L2 )-(C0-C6 alkylene)-; wherein, the C0-C 10 The hydrogen in the alkylene group may optionally be substituted by one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, -(C0-C6 alkylene)-(C3-C6 alkylene) 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 Alkyl), -O(C) 0- 10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 Alkyl); wherein, R L2 Selected from: H, deuterium, halogen, cyano, nitro, azide, C1-C 10 Alkyl, -(C0-C6 alkylene)-(C3-C6 alkylene) 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0- 10 alkyl), -CO(C) 0-10 alkyl); Preferably, L2 is selected from: single bonds, Preferably, L3 is selected from: single bond, -(C0-C6 alkylene)-OC(O)-, C3-C 10 Cycloalkylene, C6-C 10 arylene, 4-10 membered heterocyclic alkylene; wherein the C0-C6 alkylene, C3-C 10 Cycloalkylene, C6-C 10 The hydrogen atoms in the arylene and 4-10 membered heterocyclic groups may optionally be substituted with one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, C1-C 10 Haloalkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 Alkyl), -O(C) 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl); Preferably, L3 is selected from: single bonds, Preferably, D is selected from: -NH-, 5. The compound according to claim 1, characterized in that, L is L4 is a divalent group attached to L3, selected from: single bonds, L5 is a divalent group, selected from: single bonds C1-C6 alkylene, -N(C0-C 10 Alkyl)-, -CO-, -O-, Where s is an integer from 1 to 10; r is an integer from 1 to 10; P1, P2, P3, and P4 are either absent or amino acid residues, and at least one of P1, P2, P3, and P4 is an amino acid residue, wherein the amino acid residue is selected from: glycine residue, alanine residue, valine residue, leucine residue, isoleucine residue, methionine residue, proline residue, tryptophan residue, serine residue, tyrosine residue, cysteine ​​residue, phenylalanine residue, asparagine residue, glutamine residue, threonine residue, aspartic acid residue, glutamic acid residue, lysine residue, arginine residue, histidine residue, citrulline residue, ornithine residue, cysteine ​​residue, selenocysteine, hydroxyproline, hydroxylysine, and theanine; Preferably, P1, P2, P3, and P4 are independently absent or selected from: glycine residues, L-alanine residues, D-alanine residues, L-valine residues, D-valine residues, L-phenylalanine residues, D-phenylalanine residues, L-citrulline residues, D-citrulline residues, L-asparagine residues, and D-asparagine residues. L6 is selected from: Where q is an integer from 1 to 10; R L3 R L6 R L7 Independently selected from: H, C1-C 10 Alkyl, -(C0-C6 alkylene)-(C6-C 10 (Aromatic); Preferably, L is selected from: Where q is an integer from 1 to 10; Preferably, L is 6. The compound according to any one of claims 1-5, characterized in that, The compound has the following structure:

7. A conjugate or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, or deuterated thereof, said conjugate having the following structure: in, U is selected from: C0-C 10 Alkyl, C3-C 10 cycloalkyl, C6-C 10 Aryl, 4-10 membered heterocyclic groups; V is selected from: -O-, -S-, B is The C ring is a divalent group attached to the A ring; it either does not exist or is selected from: C3-C 10 Cycloalkylene, C6-C 10 arylene, 4-10 membered heterocyclic arylene; X, Y, and Z are independently selected from: single bonds, C1-C 10 Alkylene, -(C0-C6 alkylene)-O-, -(C0-C6 alkylene)-S-, -(C0-C6 alkylene)-C(O)-, -(C0-C6 alkylene)-C(S)-, -(C0-C6 alkylene)-N(R) B1 )-、-(C0-C6 alkylene)-CON(R B1 )-、-(C0-C6 alkylene)-N(R B1 CO-, -(C0-C6 alkylene)-SO2-, -(C0-C6 alkylene)-SO-, -(C0-C6 alkylene)-(C3-C 10 Cycloalkylene)-, -(C0-C6 alkylene)-(C6-C 10 arylene)-, -(C0-C6 alkylene)-(4-10 heterocyclic)-; wherein, the C0-C 10 Alkylene, C3-C 10 Cycloalkylene, C6-C 10 The hydrogen atoms in the arylene and 4-10 membered heterocyclic groups may optionally be substituted with one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, -(C0-C6 alkylene)-(C3-C 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, C1-C 10 Halogenated alkoxy groups, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)CON(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -O(C) 0-10 alkyl), -S(C 0-10 Alkyl), -SO(C) 0-10 alkyl), -SO2(C 0-10 alkyl), -SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0- 10 alkyl); Ring A is selected from: C3-C 10 Cycloalkylene, C6-C 10 arylene, 4-10 membered heterocyclic; of which, C3-C 10 Cycloalkylene, C6-C 10 The hydrogen atoms in the arylene and 4-10 membered heterocyclic groups may optionally be substituted with one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, -(C0-C6 alkylene)-(C3-C 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, C1-C 10 Haloalkoxy, -(C0-C6 alkylene)-N(C 0-10 Alkyl)(C 0-10 Alkyl), -(C0-C6 alkylene)-N(C 0-10 Alkyl)CO(C 0-10 Alkyl), -(C0-C6 alkylene)-N(C 0-10 Alkyl)CON(C 0-10 Alkyl), -(C0-C6 alkylene)-N(C 0-10 Alkyl)SO2(C 0-10 Alkyl), -(C0-C6 alkylene)-O(C 0-10 Alkyl), -(C0-C6 alkylene)-S(C 0-10 Alkyl), -(C0-C6 alkylene)-SO(C 0-10 Alkyl), -(C0-C6 alkylene)-SO2(C 0-10 Alkyl), -(C0-C6 alkylene)-SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -(C0-C6 alkylene)-COO(C 0-10 Alkyl), -(C0-C6 alkylene)-OCO(C 0-10 Alkyl), -(C0-C6 alkylene)-CON(C 0-10 Alkyl)(C 0-10 Alkyl), -(C0-C6 alkylene)-CO(C 0-10 alkyl); D is L1, L2, and L3 are independently selected from: single bonds, C1-C 10 Alkylene, -O-(C0-C6 alkylene)-, -S-(C0-C6 alkylene)-, -N(R L0 )-(C0-C6 alkylene)-, -N(R L0 )C(O)-(C0-C6 alkylene)-、-N(R L0 )C(O)O-(C0-C6 alkylene)-、-N(R L0 )C(O)N(R L0 -(C0-C6 alkylene)-, -C(O)-(C0-C6 alkylene)-, -C(S)-(C0-C6 alkylene)-, -C(O)O-(C0-C6 alkylene)-, -OC(O)-(C0-C6 alkylene)-, -(C0-C6 alkylene)-OC(O)-, -CON(R L0 -(C0-C6 alkylene)-, -SO2-(C0-C6 alkylene)-, -SO-(C0-C6 alkylene)-, C3-C 10 Cycloalkylene, C6-C 10 arylene, 4-10 membered heterocyclic alkylene; wherein, the CO-C 10 Alkylene, C3-C 10 Cycloalkylene, C6-C 10 The hydrogen atoms in the arylene and 4-10 membered heterocyclic groups may optionally be substituted with one or more groups selected from the following: deuterium, halogen, cyano, nitro, azide, C1-C. 10 Alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, -(C0-C6 alkylene)-(C3-C 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, C1-C 10 Halogenated alkoxy groups, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)CON(C 0-10 alkyl), -N(C) 0- 10 Alkyl)SO2(C 0-10 Alkyl), -O(C) 0-10 alkyl), -S(C 0-10 Alkyl), -SO(C) 0-10 alkyl), -SO2(C 0-10 alkyl), -SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl); R V1 R V2 R B1 R L0 Independently selected from: H, deuterium, halogen, cyano, nitro, azide, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, -(C0-C6 alkylene)-(C3-C 10 cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic), C1-C 10 Haloalkyl, C1-C 10 Halogenated alkoxy groups, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)CON(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -O(C) 0-10 alkyl), -S(C 0-10 Alkyl), -SO(C) 0-10 alkyl), -SO2(C 0-10 alkyl), -SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -CO(C) 0-10 alkyl); n is an integer or decimal between 1 and 10; L' is the connector; Bm is the connecting part.

8. The conjugate according to claim 7, characterized in that, L' is L4 is a divalent group attached to L3, selected from: single bonds, L5 is a divalent group, selected from: single bonds C1-C6 alkylene, -N(C0-C 10 Alkyl)-, -CO-, -O-, Where s is an integer from 1 to 10; r is an integer from 1 to 10; P1, P2, P3, and P4 are either absent or amino acid residues, and at least one of P1, P2, P3, and P4 is an amino acid residue, wherein the amino acid residue is selected from: glycine residue, alanine residue, valine residue, leucine residue, isoleucine residue, methionine residue, proline residue, tryptophan residue, serine residue, tyrosine residue, cysteine ​​residue, phenylalanine residue, asparagine residue, glutamine residue, threonine residue, aspartic acid residue, glutamic acid residue, lysine residue, arginine residue, histidine residue, citrulline residue, ornithine residue, cysteine ​​residue, selenocysteine, hydroxyproline, hydroxylysine, and theanine; L6' is selected from: Where q is an integer from 1 to 10; R L3 Selected from: H, C1-C 10 Alkyl, -(C0-C6 alkylene)-(C6-C 10 (Aromatic); Preferably, L is selected from: q is an integer from 1 to 10; Preferably, the binding portion is an antibody, an antibody fragment, or an antigen-binding fragment; Preferably, the protein bound to the binding portion is a surface antigen; Preferably, the surface antigen is selected from: 5T4, ACE, ADRB3, AKAP-4, ALK, androgen receptor, AOC3, APP, axonin 1, AXL, B7H3, B7-H4, BCL2, BCMA, bcr-abl, BORIS, BST2, C242, C4.4a, CA 125, CA6, CA9, CAIX, CCL11, CCR5, CD123, CD133, CD138, CD142, CD15, CD15-3, CD171, CD179a, CD18, CD19, CD19-9, CD2, CD20, CD22, CD23, CD24, CD25, CD27L, CD28, CD3, CD30, CD31, CD300LF, CD33, CD352, CD37, CD38, CD4, CD40, CD41, CD44, CD44v6, CD5 CD51, CD52, CD54, CD56, CD62E, CD62P, CD62L, CD70, CD71, CD72, CD74, CD79a, CD79b, CD80, CD90, CD97, CD125, CD138, CD141, CD147, CD152, CD154, CD326, CEA, CEACAM5, CFTR, condensation factor, cKit, closure protein 3, closure protein 18.2, CLDN6, CLEC12A, CLL-1, cll3, c-MET, Crypto 1. Growth factors, CS1, CTLA-4, CXCR2, CXORF61, cyclin B1, CYP1B1, cadherin-3, cadherin-6, DLL3, E7, EDNRB, EFNA4, EGFR, EGFRvIII, ELF2M, EMR2, ENPP3, EPCAM, EphA2, liver glycoside A4, liver glycoside B2, EPHB4, ERBB2 (Her2 / neu), ErbB3, ERG (TMPRSS2ETS fusion gene), E TBR, ETV6-AML, FAP, FCAR, FCRL5, FGFR1, FGFR2, FGFR3, FGFR4, FLT3, folate receptor α, folate receptor β, FOLR1, Fos-associated antigen 1, fucose GM1, GCC, GD2, GD3, GloboH, GM3, GPC1, GPC2, GPC3, gp1OO, GPNMB, GPR20, GPRC5D, GUCY2C, HAVCR1, HER2, HER3, HGF, HMI.

24. HMWMAA, HPV E6, hTERT, human telomerase reverse transcriptase, ICAM, ICOS-L, IFN-α, IFNγ, IGF-I receptor, IGLL1, IL-2 receptor, IL-4 receptor, IL-13Ra2, IL-11Ra, IL-1, IL-12, IL-23, IL13, IL-22, IL-4, IL-5, IL-6, interferon receptor, integrins (including α4, α). v β3, α v β5, α v β6, α1β4, α4β1, α4β7, α5β1, α6β4, α IIb β3 integrin), integrin αV, intestinal carboxyl esterase, KIT, LAGE-1a, LAIR1, LAMP-1, LCK, Legumain, LewisY, LFA-1 (CD11a), L-selectin (CD62L), LILRA2, LIV-1, LMP2, LRRC15, LY6E, LY6K, LY75, MAD-CT-1, MAD-CT-2, MAGE A1, MelanA / MART1, mesothelin, ML-IAP, MSLN, mucin, MUC1, MUC16, mut hsp70-2, MYCN, myostatin, NA17, NaPi2b, NCA-90, NCAM, Nectin-4, NGF, NOTCH1, NOTCH2, NOTCH3, NOTCH4, NY-BR-1, NY-ESO-1, o-acetyl-GD2, OR51E2, OY-TES1, p53, p53 mutant, PANX3, PAP, PAX3, PAX5, p-CAD, PCTA-1 / galactagoguerin 8, PD-L1, PD-L2, PDGFR, PDGFR-β, phosphatidylserine, PIK3CA, PLAC1, polysialic acid, prostate enzymes, prostate cancer cells, prostate Adenosine, Pseudomonas aeruginosa, rabies virus, survivin and telomerase, PRSS21, PSCA, PSMA, PTK7, RAGE-1, RANKL, Ras mutant, respiratory syncytial virus, rhesus monkey factor, RhoC, RON, ROR1, ROR2, RU1, RU2, sarcoma translocation breakpoint, SART3, SLAMF7, SLC44A4, sLe, SLITRK6, spermin 17, sphingosine 1-phosphate, SSEA-4, SSX2, STEAP1, TAG72, TARP, TCRβ, TEM1 / CD248, TEM7R, tendinin C, TF, TGF-1, TGF-β2, TNF-α, TGS5, Tie 2. One or more of the following: TIM-1, TnAg, TRAC, TRAIL-R1, TRAIL-R2, TROP-2, TRP-2, TRPV1, TSHR, tumor antigen CTAA16.88, tyrosinase, UPK2, VEGF, VEGFR1, VEGFR2, vimentin, WT1, XAGE1, B7H3, LAG-3 (CD223), PD-1 / PD-L1, BTLA, TIM-3, AA2R, CEACAM1, SIRPα, CD200R, c-met; More preferably, the surface antigen is selected from: HER2, B7H3, CD20, CD38, CD33, BCMA, CD138, EGFR, FGFR4, GD2, PDGFR, TEM1 / CD248, TROP-2, PD-L1, CD123, c-met; Preferably, the antibody is selected from: rituximab, trastuzumab, gemtuzumab, pertuzumab, obitutuzumab, oflamuzumab, olatuzumab, antuximab, ixartuzumab, saxituzumab, U3-1784, daratumumab, STI-6129, OR000213, lintuzumab, huMy9-6, belantanumab, indextuzumab, cetuximab, dinutuximab, anti-CD38 A2 antibody, HuaT13 / 5 antibody, alemtuzumab, teimomab, tosimob, bevacizumab, panitumumab, trimemumab, teimomab, caputuzumab, ovovacuumab, vetouzumab, and anti-B7-H3 antibody.

9. The conjugate according to claim 7 or 8, characterized in that, The conjugate is selected from the following structures:

10. The conjugate according to claim 9, characterized in that, The conjugate has the structure shown in L-1, wherein the Bm portion is Herceptin, n = 5-6; or, The conjugate has the structure shown in L-4, wherein the Bm portion is Herceptin, n = 3-4, 5-6, 7-8, 8-9; or, The conjugate has the structure shown in L-4, wherein the Bm portion is Ifinatamab, n = 3-4, 6-7; or, The conjugate has the structure shown in L-3, wherein the Bm portion is Herceptin, n = 3-4, 4-5, 7-8; or, The conjugate has the structure shown in L-5, wherein the Bm portion is Herceptin, n = 2-3; or, The conjugate has the structure shown in L-6, wherein the Bm portion is Herceptin, n = 1-3; or, The conjugate has the structure shown in L-52, wherein the Bm portion is Herceptin, n = 3-4; or, The conjugate has the structure shown in L-53, wherein the Bm portion is Herceptin, n = 3-4; or, The conjugate has the structure shown in L-54, wherein the Bm portion is Herceptin, n = 3-4; or, The conjugate has the structure shown in L-55, wherein the Bm portion is Herceptin and n = 2-3.

11. A pharmaceutical composition comprising the compound of any one of claims 1-6 or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, or deuterated thereof, or the conjugate of any one of claims 7-10 or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, or deuterated thereof, and one or more pharmaceutically acceptable excipients.

12. The use of the compound of any one of claims 1-6 or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate or deuterated thereof, or the conjugate of any one of claims 7-10 or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate or deuterated thereof, or the pharmaceutical composition of claim 11 in the preparation of a medicament for the prevention and / or treatment of a disease; Preferably, the disease is selected from: tumors, cardiovascular and cerebrovascular diseases, and viral infection-related diseases; Preferably, the tumor is selected from: liver cancer, lung cancer, stomach cancer, breast cancer, colon cancer, bile duct cancer, bladder cancer, head and neck cancer, cervical cancer, ovarian cancer, prostate cancer, thyroid cancer, squamous cell carcinoma, lymphoma, sarcoma, acute myeloid leukemia, chronic lymphocytic leukemia, multiple myeloma, and myelodysplastic syndrome.

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