Polypeptide, bioactive conjugate as well as preparation method and application of polypeptide and bioactive conjugate

Abstract

The invention provides a bioactive conjugate as well as a preparation method and application thereof, and particularly relates to a bioactive conjugate using a linker of a polypeptide unit (-P1-P2-) and application of the bioactive conjugate in the field of disease treatment, including but not limited to treatment of tumor diseases. In addition, the invention also provides a linker-drug, a linker and a polypeptide fragment which are used by the bioactive conjugate as well as a preparation method and application of the linker-drug, the linker and the polypeptide fragment.

Description

Polypeptides, bioactive conjugates, and preparation methods and applications thereof Technical Field

[0001] The present application relates to the field of biomedicine, and specifically to a polypeptide, a linker, a linker drug, a bioactive conjugate, a preparation method thereof, and applications in preventing / treating diseases. Background Art

[0002] Abnormal cell activity can lead to various diseases in the body, such as cancer, autoimmune diseases, neurodegenerative diseases, etc. Cancer is manifested as abnormal cell proliferation. Intervention against abnormal cell proliferation is a conventional method for cancer treatment, and the use of small chemical molecules to treat cancer is currently a widely used method. However, many cytotoxic drug molecules are limited in their clinical use due to problems such as lack of target selectivity, unsatisfactory pharmacokinetic properties, or excessive toxicity. As a result, bioactive conjugates have emerged. Bioactive conjugates have played a huge role in improving targeting and solving the unsatisfactory properties of the drug itself. Among them, antibody-drug conjugates (ADCs) have become the leader of bioactive conjugates in tumor treatment. As of September 2023, a total of 13 ADCs are being used in clinical treatment worldwide: Mylotarg (targeting CD33), Adcetris (targeting CD30), Kadcyla (targeting Her2), Besponsa (targeting CD33), Polivy (targeting CD79b), Padcev (targeting Nectin-4), Enhertu (targeting Her2), Trodelvy (targeting Trop2), Zynlonta (targeting CD19), Tivdak (targeting TF), Elahere (targeting FRα), Akalux (targeting EGFR), and Aidixi (targeting Her2). In 2022 alone, 249 new ADC-related clinical trials were initiated. The ADCs in clinical trials not only cover the targets of marketed drugs, but also involve several new potential ADC targets, such as B7-H3, B7-H4, ROR1, 5T4, ITGB6, LIV-1, and others.

[0003] ADCs consist of three components: an antibody, a linker, and a drug payload. This includes the connection between the antibody and the linker, and the connection between the linker and the drug payload. The actual active component of an ADC is the drug payload. Its mechanism of action is to utilize the specific binding of the antibody and antigen to concentrate the drug on the surface of tumor cells, followed by a specific mechanism for drug release at the tumor site. Release mechanisms include external release from tumor cells, leveraging the specific environment of tumor tissue, or utilizing the endocytosis of tumor cells to internalize the ADC into lysosomes. The active drug molecules are then released under the action of various enzymes within the lysosomes, thereby exerting their biological functions, such as inducing apoptosis and achieving anti-tumor effects. Therefore, the key to ADC drug development lies in, on the one hand, maintaining the stability of the ADC in the body's general environment, such as in the plasma circulation, and, on the other hand, ensuring that the ADC is released quickly and efficiently after reaching the disease site.

[0004] In terms of environmental stability, the choice of ADC linker and the method of linking the linker to the antibody often play a key role. Since the connection between the antibody and the linker usually needs to be carried out in a predominantly aqueous solution, the available reaction methods are relatively limited. For example, the marketed ADCs Mylotarg, Besponsa, Kadcyla, Akalux, and Elahere utilize the reaction of amino groups on the lysine residues on the antibody surface with activated esters on the linker to achieve antibody extension. Due to their good nucleophilic properties, the amino groups on the lysine residues can also react with various functional groups such as o-phthalaldehyde. Other marketed ADCs all utilize the reduction of interchain disulfide bonds between the antibody light-heavy chain and heavy-heavy chain to obtain free sulfhydryl groups on cysteine ​​residues, and then Michael addition of the free sulfhydryl groups with maleimide to achieve antibody extension. Due to the stronger nucleophilic properties of sulfhydryl groups, more functional groups can react with sulfhydryl groups in aqueous reaction systems, resulting in a wider range of methods for utilizing free sulfhydryl groups to achieve antibody extension. In addition, there are other methods to achieve the extension of antibodies to linkers, such as using amino acid point mutations to modify the antibody backbone to obtain cysteine ​​or non-natural amino acids, and then using the sulfhydryl group on the modified cysteine ​​residue or the ketone group or azide group on the non-natural amino acid residue to react with the functional group on the linker; or first extending the antibody polypeptide, and then using the functional group on the extended polypeptide to react with the linker; or performing glycosylation modification on the polypeptide, and using the functional group in the modified sugar group, usually a ketone group, an azide group or an alkyne group, to react with the linker; or using the catalytic action of an enzyme to enzymatically link the antibody or the antibody extended along the polypeptide to the corresponding enzyme substrate linker.

[0005] The choice of linker structure often balances the stability of the ADC in the environment and its release at the tumor site. Stable linkers are one option, such as Kadcyla, which uses a stable linker. This type of linker can ensure the stability of the ADC in the environment. However, the disadvantage is that ADCs using this type of linker can affect the release efficiency of the drug, and the released drug often retains a portion of the linker. This not only limits the choice of drug, but also restricts the application of the ADC because the released drug often lacks a bystander effect. In contrast, the use of linkers that respond to lysosomal enzymes partially addresses this issue. Because lysosomal enzymes are largely absent outside the cell, ADCs using this type of linker can still maintain considerable stability in the body's environment. After endocytosis and lysosomal transport, they can be efficiently released in the lysosome, thereby exerting the biological function of the drug. Furthermore, this release mechanism often allows the drug to be released intact. Since the drug often has cell penetrating ability, ADCs prepared using this type of linker often exhibit a bystander effect, thereby better exerting anti-tumor effects. A typical example of this type of linker ADC is Enhertu. While using the same antibody as Kadcyla, Enhertu has better therapeutic effects than Kadcyla and is applicable to a wider range of disease populations. However, this type of linker also has shortcomings, namely, it is highly dependent on endocytosis and lysosomal transport. Although the drug can be efficiently released after the ADC enters the lysosome, the actual drug release efficiency of this type of ADC after reaching the tumor tissue is not high due to the influence of endocytosis efficiency and lysosomal transport efficiency. For antigen targets with low endocytosis efficiency, this type of linker is basically unusable. If a linker that does not rely on endocytosis can be found, the drug release efficiency can be further improved. In this regard, Gilead's Trodelvy has made corresponding attempts. The CL2A-SN38 linker drug used in Trodelvy can release the drug-loaded SN38 in the environment to exert anti-tumor activity. However, in order to achieve the purpose of extracellular release, the linker sacrifices the stability of ADC in the body environment. The half-life of Trodelvy in plasma is less than 24 hours, resulting in serious inconsistency in the pharmacokinetic properties of ADC and naked antibodies. The pharmacokinetic deficiencies can only be compensated by increasing the frequency of clinical injections, which has a certain impact on the patient experience of Trodelvy. The blood toxicity caused by Trodelvy's instability in plasma has also become the main reason for the clinical dose limit of Trodelvy.

[0006] Therefore, developing linkers that can maintain relative stability in plasma and efficiently release drug-loaded materials in the tumor microenvironment and / or lysosomes is of great significance for further improving the therapeutic effects of bioactive conjugates and enhancing patients' medication experience.

[0007] Tissue Factor (TF), also known as coagulation factor III, is a single-pass transmembrane protein. TF is expressed at certain levels in cells of the vascular wall and somatic cells, where it plays a role in hemostasis. Studies have shown that TF is significantly overexpressed in various malignancies, such as cervical cancer, ovarian cancer, prostate cancer, breast cancer, non-small cell lung cancer, colon cancer, and pancreatic cancer, and that high TF expression is associated with a poor prognosis in many tumors. Because TF itself lacks an intracellular structure and does not play a role in intracellular signaling, developing monoclonal antibody drugs targeting TF is not a promising strategy. However, the high internalization efficiency of TF protein after binding to antibodies provides a promising direction for the development of ADCs targeting TF. Tivdak, an approved drug targeting TF, is an ADC approved for the treatment of refractory or recurrent cervical cancer. Compared to conventional ADC development, the challenge of developing ADCs targeting TF lies in the need to balance the antibody's ability to bind to the TF antigen and the effects of the anti-TF antibody itself on coagulation.

[0008] Summary of the Invention

[0009] To improve the therapeutic efficacy of antibody-drug conjugates (ADCs) or other ligand-drug conjugates, reduce drug toxicity and side effects, and increase the therapeutic window, it is crucial to identify linkers that maintain relative stability in plasma while efficiently releasing the drug in the tumor microenvironment and / or lysosomes. This application describes a series of peptides that respond to both tumor tissue-specific MMP-2 and / or MMP-9 and Cathepsin B in tumor cells, as well as spacer structures that connect the peptides to the antibody and payload.

[0010] On the one hand, the present application provides a polypeptide or polypeptide fragment having a structure shown as -P1-P2-, wherein polypeptide P1 and polypeptide P2 respond to enzymes respectively.

[0011] In some embodiments, P1 comprises a polypeptide responsive to matrix metalloproteinase 2 (MMP-2) or matrix metalloproteinase 9 (MMP-9) or cathepsin B, and P2 comprises a polypeptide responsive to cathepsin B or matrix metalloproteinase 2 (MMP-2) or matrix metalloproteinase 9 (MMP-9).

[0012] In some embodiments, the P1 comprises the following polypeptide structure: -X1-P-X2-X3-Z-, and the P2 comprises the following polypeptide structure: -X1'-X2'-X3'-X4'-; wherein, P is a proline residue, X1, X2, X3, Z, X1', X2', X3', X4' represent absence, an amino acid or an amino acid derivative, and at least 2 of X1', X2', X3' and X4' are single amino acids or single amino acid derivatives.

[0013] In some embodiments, the P1 and P2 are responsive to matrix metalloproteinase 2 (MMP-2) and / or matrix metalloproteinase 9 (MMP-9), and cathepsin B.

[0014] In some embodiments, X1 represents absence, a single amino acid, or a single amino acid derivative, X2, X3, and Z represent a single amino acid or a single amino acid derivative, X1' and X2' represent a single amino acid or a single amino acid derivative, and X3' and X4' represent absence or a single amino acid or a single amino acid derivative.

[0015] In some embodiments, the X1 is selected from the group consisting of: absent, Gly (G), Val (V), His (H), Ser (S), Ile (I), the X2 is selected from the group consisting of: Ala (A), Ser (S), Leu (L), Arg (R), Gln (Q), Val (V), Cit, the X3 is selected from the group consisting of: Gly (G), Ser (S), Pro (P), Asn (N), Ala (A), the Z is selected from the group consisting of: Leu (L), Ile (I), Val (V), Phe (F), Met (M), Ala (A), Trp(W), Gln(Q), N,N-dimethyl-3-aminopropionic acid (Dap(Me2)), N,N-dimethyl-4-aminobutyric acid (Dab(Me2)), N,N-dimethylornithine (Orn(Me2)), N,N-dimethyllysine (Lys(Me2)), N,N-diethyl-3-aminopropionic acid (Dap(Et2)), N,N-diethyl-4-aminobutyric acid (Dab(Et2)), N,N-diethylornithine (Orn(Et2)), N,N-diethyllysine (Lys(Et2)).

[0016] In some embodiments, the X1' is selected from the group consisting of Val (V), Als (A), Leu (L), Ile (I), Met (M), Phe (F), Trp (W), Pro (P), Tyr (Y), Gly (G), and Glu (E), and the X2' is selected from the group consisting of Val (V), Leu (L), Ile (I), Met (M), Trp (W), Pro (P), Lys (K), Lys (Me2), Lys (Et2), His (H), A Ala (A), Arg (R), Gly (G), Phe (F), Cit, Orn, Orn (Me2), Orn (Et2), wherein X3' is selected from the group consisting of: absence, Phe (F), Gly (G), Lys (K), Lys (Me2), Lys (Et2), His (H), Ala (A), Arg (R), Cit, Orn, Orn (Me2), Orn (Et2), wherein X4' is selected from the group consisting of: absence, Gly (G), Phe (F).

[0017] In some embodiments, the X1 is selected from: absent, Gly (G), Val (V), His (H), Ser (S), Ile (I), the X2 is selected from: Ala (A), Ser (S), Leu (L), Arg (R), Gln (Q), Val (V), Cit, the X3 is selected from: Gly (G), Ser (S), Pro (P), Asn (N), Ala (A), the Z is selected from: Leu (L), Ile (I), Val (V), Phe (F), M et(M), Ala(A), Trp(W), Gln(Q), N,N-dimethyl-3-aminopropionic acid (Dap(Me2)), N,N-dimethyl-4-aminobutyric acid (Dab(Me2)), N,N-dimethylornithine (Orn(Me2)), N,N-dimethyllysine (Lys(Me2)), N,N-diethyl-3-aminopropionic acid (Dap(Et2)), N,N-diethyl-4-aminobutyric acid (Dab(Et2)), N,N-diethylornithine (Orn(Et 2)), N,N-diethyllysine (Lys(Et2)), wherein X1' is selected from the group consisting of Val(V), Als(A), Leu(L), Ile(I), Met(M), Phe(F), Trp(W), Pro(P), Tyr(Y), Gly(G), Glu(E), and X2' is selected from the group consisting of Val(V), Leu(L), Ile(I), Met(M), Trp(W), Pro(P), Lys(K), Lys(Me2), Lys(Et2), His (H), Ala (A), Arg (R), Gly (G), Phe (F), Cit, Orn, Orn (Me2), Orn (Et2), wherein X3' is selected from the group consisting of: absence, Phe (F), Gly (G), Lys (K), Lys (Me2), Lys (Et2), His (H), Ala (A), Arg (R), Cit, Orn, Orn (Me2), Orn (Et2), and X4' is selected from the group consisting of: absence, Gly (G), Phe (F).

[0018] In some preferred embodiments, the X1 is absent or is Gly (G), Ser (S) or His (H); the X2 is Ala (A), Leu (L), Arg (R), Ser (S), Leu (L) or Cit; the X3 is Gly (G), Ser (S), Ala (A), Asn (N) or Pro (P); and the Z is Leu (L) or Dap (Me2).

[0019] In some preferred embodiments, X1' is Val (V) or Gly (G), X2' is Gly (G) or Cit, and at least one of X3' and X4' is absent or is Phe (F) or Gly (G). In some preferred embodiments, both X3' and X4' are absent. In some preferred embodiments, X3' is Phe (F) and X4' is Gly (G).

[0020] In some embodiments of the polypeptide or polypeptide fragment, the (-X1'-X2'-X3'-X4'-) of P2 comprises the following fragments: -Val-Cit-(V-Cit), -Ala-Lys-(AK) or -Gly-Gly-Phe-Gly-(GGFG). In some preferred embodiments, the -X1'-X2'-X3'-X4'- is -V-Cit- or -GGFG-.

[0021] In some embodiments, any of the polypeptides or polypeptide fragments is selected from:

[0022] -His-Val-Leu-Asn-Leu-Val-Cit-(SEQ ID NO: 1);

[0023] -Val-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:2);

[0024] -Ile-Pro-Val-Ser-Leu-Val-Cit-(SEQ ID NO:3);

[0025] -Gly-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:4);

[0026] -Gly-Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:5);

[0027] -Gly-Pro-Ala-Gly-Leu-Val-Cit-(SEQ ID NO: 6);

[0028] -Gly-Pro-Gln-Gly-Leu-Val-Cit-(SEQ ID NO:7);

[0029] -Gly-Pro-Leu-Gly-Leu-Val-Cit-(SEQ ID NO:8);

[0030] -Gly-Pro-Ser-Gly-Leu-Val-Cit-(SEQ ID NO:9);

[0031] -Gly-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:10);

[0032] -Gly-Pro-Ala-Ala-Leu-Val-Cit-(SEQ ID NO:11);

[0033] -Gly-Pro-Ser-Ala-Leu-Val-Cit-(SEQ ID NO:12);

[0034] -His-Val-Leu-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:13);

[0035] -Val-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:14);

[0036] -Ile-Pro-Val-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:15);

[0037] -Gly-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:16);

[0038] -Gly-Pro-Ala-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:17);

[0039] -Gly-Pro-Ala-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:18);

[0040] -Gly-Pro-Gln-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:19);

[0041] -Gly-Pro-Leu-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:20);

[0042] -Gly-Pro-Ser-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:21);

[0043] -Gly-Pro-Arg-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:22);

[0044] -Gly-Pro-Ala-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:23);

[0045] -Gly-Pro-Ser-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:24);

[0046] -Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:25);

[0047] -Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:26);

[0048] -Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:27);

[0049] -Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:28);

[0050] -Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:29);

[0051] -Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:30);

[0052] -Gly-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:31);

[0053] -Gly-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:32);

[0054] -Gly-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:33);

[0055] -Gly-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:34);

[0056] -Gly-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:35);

[0057] -His-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:36);

[0058] -His-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:37);

[0059] -His-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:38);

[0060] -His-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:39);

[0061] -His-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:40);

[0062] -His-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:41);

[0063] -Ser-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:42);

[0064] -Ser-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:43);

[0065] -Ser-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:44);

[0066] -Ser-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:45);

[0067] -Ser-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:46);

[0068] -Ser-Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:47);

[0069] -Ser-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:48);

[0070] -Gly-Pro-Ala-Pro-Leu-Val-Cit-(SEQ ID NO:49);

[0071] -Gly-Pro-Ala-Asn-Leu-Val-Cit-(SEQ ID NO:50);

[0072] -Gly-Pro-Arg-Gly-Dap(Me2)-Val-Cit-(SEQ ID NO:51);

[0073] -Gly-Pro-Arg-Asn-Dap(Me2)-Val-Cit-(SEQ ID NO:60);

[0074] -Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:61);

[0075] -Gly-Pro-Arg-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO: 62);

[0076] -Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:63);

[0077] -Gly-Pro-Cit-Asn-Leu-Val-Cit-(SEQ ID NO: 64);

[0078] -Ser-Pro-Cit-Ser-Dap(Me2)-Val-Cit-(SEQ ID NO:65);

[0079] -Pro-Arg-Ser-Dap(Me2)-Val-Cit-(SEQ ID NO:66);

[0080] -Pro-Cit-Asn-Leu-Val-Cit-(SEQ ID NO:67);

[0081] -Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:68);

[0082] Or a variant having one or two mismatches relative to the sequence of any one of SEQ ID NOs: 1-51 and 60-68.

[0083] In some embodiments, the polypeptide or polypeptide fragment is selected from:

[0084] -GPLSLVCit-(SEQ ID NO:4);

[0085] -GPASLVCit-(SEQ ID NO:5);

[0086] -GPAGLVCit-(SEQ ID NO:6);

[0087] -GPRGLVCit-(SEQ ID NO:10);

[0088] -GPSALVCit-(SEQ ID NO:12);

[0089] -GPLGLGGFG-(SEQ ID NO:20);

[0090] -GPRGLGGFG-(SEQ ID NO:22);

[0091] -GPAALGGFG-(SEQ ID NO:23);

[0092] -PRGLVCit-(SEQ ID NO:25);

[0093] -PRSLVCit-(SEQ ID NO:26);

[0094] -PRNLVCit-(SEQ ID NO:27);

[0095] -PRPLVCit-(SEQ ID NO:28);

[0096] -PCitGLVCit-(SEQ ID NO:29);

[0097] -PCitPLVCit-(SEQ ID NO:30);

[0098] -GPRSLVCit-(SEQ ID NO:31);

[0099] -GPRPLVCit-(SEQ ID NO:32);

[0100] -GPRNLVCit-(SEQ ID NO:33);

[0101] -GPCitGLVCit-(SEQ ID NO:34);

[0102] -GPCitPLVCit-(SEQ ID NO:35);

[0103] -HPRGLVCit-(SEQ ID NO:36);

[0104] -HPCitPLVCit-(SEQ ID NO:41);

[0105] -SPRGLVCit-(SEQ ID NO:42);

[0106] -SPRSLVCit- (SEQ ID NO: 43);

[0107] -SPRPLVCit- (SEQ ID NO: 44);

[0108] -SPRNLVCit- (SEQ ID NO: 45);

[0109] -SPCitGLVCit-(SEQ ID NO:46);

[0110] -SPCitSLVCit-(SEQ ID NO:47);

[0111] -SPCitPLVCit-(SEQ ID NO:48);

[0112] -GPAPLVCit- (SEQ ID NO: 49);

[0113] -GPANLVCit- (SEQ ID NO: 50);

[0114] -GPRNDap(Me2)VCit-(SEQ ID NO:60);

[0115] -PLSLVCit- (SEQ ID NO: 61);

[0116] -GPRNLGGFG- (SEQ ID NO: 62);

[0117] -PCitSLVCit- (SEQ ID NO: 63);

[0118] -GPCitNLVCit-(SEQ ID NO:64);

[0119] -SPCitSDap(Me2)VCit-(SEQ ID NO:65);

[0120] -PRSDap(Me2)VCit-(SEQ ID NO:66);

[0121] -PCitNLVCit- (SEQ ID NO: 67);

[0122] -PASLVCit- (SEQ ID NO: 68);

[0123] or variants having one or two mismatches relative to any of the above sequences.

[0124] In some embodiments, the polypeptide or polypeptide fragment is selected from:

[0125] -Gly-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:4);

[0126] -Gly-Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:5);

[0127] -Gly-Pro-Ala-Gly-Leu-Val-Cit-(SEQ ID NO:6);

[0128] -Gly-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:10);

[0129] -Gly-Pro-Ser-Ala-Leu-Val-Cit-(SEQ ID NO:12);

[0130] -Gly-Pro-Leu-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:20);

[0131] -Gly-Pro-Arg-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:22);

[0132] -Gly-Pro-Ala-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:23);

[0133] -Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:25);

[0134] -Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:26);

[0135] -Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:27);

[0136] -Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:28);

[0137] -Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:29);

[0138] -Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:30);

[0139] -Gly-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:31);

[0140] -Gly-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:32);

[0141] -Gly-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:33);

[0142] -Gly-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:34);

[0143] -Gly-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:35);

[0144] -His-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:36);

[0145] -Ser-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:42);

[0146] -Ser-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:43);

[0147] -Ser-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:44);

[0148] -Ser-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:45);

[0149] -Ser-Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:47);

[0150] -Ser-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:48);

[0151] -Gly-Pro-Ala-Asn-Leu-Val-Cit-(SEQ ID NO:50);

[0152] -Gly-Pro-Arg-Asn-Dap(Me2)-Val-Cit-(SEQ ID NO:60);

[0153] -Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:61);

[0154] -Gly-Pro-Arg-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO: 62);

[0155] -Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:63);

[0156] -Gly-Pro-Cit-Asn-Leu-Val-Cit-(SEQ ID NO: 64);

[0157] -Ser-Pro-Cit-Ser-Dap(Me2)-Val-Cit-(SEQ ID NO:65);

[0158] -Pro-Arg-Ser-Dap(Me2)-Val-Cit-(SEQ ID NO:66);

[0159] -Pro-Cit-Asn-Leu-Val-Cit-(SEQ ID NO:67);

[0160] -Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:68);

[0161] or variants having one or two mismatches relative to any of the above sequences.

[0162] In some embodiments, the polypeptide or polypeptide fragment is one or more selected from the following:

[0163] -GPASLVCit-(SEQ ID NO:5);

[0164] -GPAGLVCit-(SEQ ID NO:6);

[0165] -GPRGLVCit-(SEQ ID NO: 10);

[0166] -GPSALVCit-(SEQ ID NO: 12);

[0167] -GPLGLGGFG- (SEQ ID NO: 20);

[0168] -GPRGLGGFG- (SEQ ID NO: 22);

[0169] -GPAALGGFG-(SEQ ID NO:23);

[0170] -PRGLVCit-(SEQ ID NO:25);

[0171] -PRSLVCit-(SEQ ID NO:26);

[0172] -PRNLVCit-(SEQ ID NO:27);

[0173] -PRPLVCit-(SEQ ID NO:28);

[0174] -PCitGLVCit-(SEQ ID NO:29);

[0175] -PCitPLVCit-(SEQ ID NO:30);

[0176] -GPRSLVCit-(SEQ ID NO:31);

[0177] -GPRPLVCit-(SEQ ID NO:32);

[0178] -GPRNLVCit-(SEQ ID NO:33);

[0179] -GPCitGLVCit-(SEQ ID NO:34);

[0180] -GPCitPLVCit-(SEQ ID NO:35);

[0181] -HPRGLVCit-(SEQ ID NO:36);

[0182] -HPCitPLVCit-(SEQ ID NO:41);

[0183] -SPRGLVCit-(SEQ ID NO:42);

[0184] -SPRSLVCit-(SEQ ID NO:43);

[0185] -SPRPLVCit-(SEQ ID NO:44);

[0186] -SPRNLVCit-(SEQ ID NO:45);

[0187] -SPCitGLVCit-(SEQ ID NO:46);

[0188] -SPCitSLVCit-(SEQ ID NO:47);

[0189] -SPCitPLVCit-(SEQ ID NO:48);

[0190] -GPAPLVCit- (SEQ ID NO: 49);

[0191] -GPANLVCit- (SEQ ID NO: 50);

[0192] -GPRNDap(Me2)VCit-(SEQ ID NO:60);

[0193] -PLSLVCit- (SEQ ID NO: 61);

[0194] -GPRNLGGFG- (SEQ ID NO: 62);

[0195] -PCitSLVCit- (SEQ ID NO: 63);

[0196] -GPCitNLVCit-(SEQ ID NO:64);

[0197] -SPCitSDap(Me2)VCit-(SEQ ID NO:65);

[0198] -PRSDap(Me2)VCit-(SEQ ID NO:66);

[0199] -PCitNLVCit- (SEQ ID NO: 67);

[0200] -PASLVCit- (SEQ ID NO: 68).

[0201] In some embodiments, the polypeptide or polypeptide fragment is one or more selected from the following:

[0202] -GPAGLVCit-(SEQ ID NO:6);

[0203] -GPRGLVCit-(SEQ ID NO: 10);

[0204] -PRGLVCit-(SEQ ID NO:25);

[0205] -PRSLVCit-(SEQ ID NO:26);

[0206] -PRNLVCit-(SEQ ID NO:27);

[0207] -PRPLVCit-(SEQ ID NO:28);

[0208] -PCitGLVCit-(SEQ ID NO:29);

[0209] -PCitPLVCit-(SEQ ID NO:30);

[0210] -GPRSLVCit-(SEQ ID NO:31);

[0211] -GPRPLVCit-(SEQ ID NO:32);

[0212] -GPRNLVCit-(SEQ ID NO:33);

[0213] -GPCitGLVCit-(SEQ ID NO:34);

[0214] -GPCitPLVCit-(SEQ ID NO:35);

[0215] -HPRGLVCit-(SEQ ID NO:36);

[0216] -SPRGLVCit-(SEQ ID NO:42);

[0217] -SPRSLVCit-(SEQ ID NO:43);

[0218] -SPRPLVCit-(SEQ ID NO:44);

[0219] -SPRNLVCit-(SEQ ID NO:45);

[0220] -SPCitSLVCit-(SEQ ID NO:47);

[0221] -SPCitPLVCit-(SEQ ID NO:48);

[0222] -GPANLVCit-(SEQ ID NO:50);

[0223] -GPRNDap(Me2)VCit-(SEQ ID NO:60);

[0224] -PLSLVCit-(SEQ ID NO:61);

[0225] -GPRNLGGFG-(SEQ ID NO:62);

[0226] -PCitSLVCit- (SEQ ID NO: 63);

[0227] -GPCitNLVCit-(SEQ ID NO:64);

[0228] -SPCitSDap(Me2)VCit-(SEQ ID NO:65);

[0229] -PRSDap(Me2)VCit-(SEQ ID NO:66);

[0230] -PCitNLVCit- (SEQ ID NO: 67);

[0231] -PASLVCit- (SEQ ID NO: 68).

[0232] On the other hand, the present application provides a linker (also referred to as a "linker moiety"), wherein the linker comprises the above-mentioned polypeptide or polypeptide fragment having the structure represented by -P1-P2-.

[0233] In some embodiments, the linker has the following general formula: C1-La-L1-P1-P2-L2,

[0234] Wherein C1 is a linker, used to connect the biological ligand unit;

[0235] La is empty or a tuning spacer;

[0236] L1 is empty or a spacer;

[0237] P1-P2 are the polypeptides or polypeptide fragments described above;

[0238] L2 is empty or a spacer, used to connect a payload with a specific biological function;

[0239] The positions of La and L1 can be swapped.

[0240] In some embodiments, the biological ligand unit includes antibodies, antibody fragments, proteins, polypeptides, polynucleotides, and chemical small molecules, but is not limited thereto.

[0241] In some embodiments, in the linker, C1 is selected from the following structures:

[0242] in Indicates the connection position with the biological ligand unit, Indicates the connection position with La.

[0243] In some embodiments, the C1 comprises the following structures: -(succinimide-3-yl-N)-, -CH2-C(=O)-, Or -C(=O)-, wherein the -(succinimide-3-yl-N)- has the following structure:

[0244] in Indicates the connection position with the biological ligand unit, Indicates the connection position with La.

[0245] In some embodiments, the C1 is -(succinimid-3-yl-N)-.

[0246] In some embodiments, the La is empty or selected from the following structures or a combination of the following structures:

[0247] R1-SS-R2,

[0248] R1-(CH2CH2O) m -R2,

[0249] R1-(CH2CH2O) m -CH2CH2S-(CH2CH2O) n -R2,

[0250] R1-(CH2CH2O) m -CH2CH2NH-(CH2CH2O) n -R2,

[0251] R1-(CH2CH2O) m -CH2CH2(C=O)NH-(CH2CH2O) n -R2,

[0252] a is 0 or 1;

[0253] m is any integer from 1 to 20;

[0254] n is any integer from 1 to 20;

[0255] R3 is H, -C1-C 10 Alkylene-, -C3-C8 carbocyclic-, -C2-C 10 Heteroaryl-, -C3-C 24 Alkylheteroaryl-, wherein -C1-C 10 Alkylene-, -C3-C8 carbocyclic-, -C2-C 10 Heteroaryl-, -C3-C 24Alkylheteroaryl-can be substituted by one or more heteroatoms, which can be O, S, NR 3 , where R 3 is H or -C1-C4 alkylene;

[0256] R1 is a connecting structure with C1, and R2 is a connecting structure with L1.

[0257] In some embodiments, R1 is a single bond or is selected from the following structures:

[0258] -X 1 -C1-C 10 Alkylene-X 2 -、-X 1 -C1-C 10 Heteroalkylene-X 2 -、-X 1 -C3-C8 carbocyclyl-X 2 -、-X 1 -Arylene-X 2 -、-X 1 -C1-C 10 Alkylene-arylene-X 2 -、-X 1 -Arylene-C1-C 10 Alkylene-X 2 -、-X 1 -C1-C 10 Alkylene-(C3-C8 carbocyclyl)-X 2 -、-X 1 -(C3-C8 carbocyclyl)-CC1-C 10 Alkylene-X 2 -、-X 1 -C3-C8 heterocyclyl-X 2 -、-X 1 -C1-C 10 Alkylene-(C3-C8 heterocyclyl)-X 2 -、-X 1 -(C3-C8 heterocyclyl)-C1-C 10 Alkylene-X 2 -;

[0259] wherein X 1 is absent or selected from O, S, NH, C(=O), NHC(=O), C(=O)NH, NHC(=O)NH, X 2 is absent or selected from O, NH, S, C(═O), NHC(═O), C(═O)NH, and NHC(═O)NH.

[0260] Wherein said R1 when not a single bond, is optionally substituted by a basic unit (BU), said basic unit being -(CH2) x NH2, -(CH2) x NHR m or -(CH2) x N(R m )2; wherein x is any integer from 1 to 4; and each R m are independently selected from C1-C6 alkyl and C1-C6 haloalkyl, or two R m The groups combine with the nitrogen to which they are attached to form a 4-6 membered heterocycloalkyl ring, or an azetidinyl, pyrrolidinyl or piperidinyl group.

[0261] In some embodiments, R2 is a single bond or is selected from the following structures:

[0262] -X 3 -C1-C 10 Alkylene-X 4 -、-X 3 -C1-C 10 Heteroalkylene-X 4 -、-X 3 -C3-C8 carbocyclyl-X 4 -、-X 3 -Arylene-X 4 -、-X 3 -C1-C 10 Alkylene-arylene-X 4 -、-X 3 -Arylene-C1-C 10 Alkylene-X 4 -、-X 3 -C1-C 10 Alkylene-(C3-C8 carbocyclyl)-X 4 -、-X 3 -(C3-C8 carbocyclyl)-C1-C 10 Alkylene-X 4 -、-X 3 -C3-C8 heterocyclyl-X 4 -、-X 3 -C1-C 10 Alkylene-(C3-C8 heterocyclyl)-X 4 -、-X 3 -(C3-C8 heterocyclyl)-C1-C 10 Alkylene-X 4 -;

[0263] wherein X 3is absent or O, S, NH, C(=O), NHC(=O), C(=O)NH, NHC(=O)NH, X 4 is absent or selected from O, S, NH, C(═O), NHC(═O), C(═O)NH, and NHC(═O)NH.

[0264] Wherein said R2 when not a single bond, is optionally substituted by a basic unit (BU), said basic unit being -(CH2) x NH2, -(CH2) x NHR m or -(CH2) x N(R m )2; wherein x is any integer from 1 to 4; and each R m are independently selected from C1-C6 alkyl and C1-C6 haloalkyl, or two R m The groups combine with the nitrogen to which they are attached to form a 4-6 membered heterocycloalkyl ring, or an azetidinyl, pyrrolidinyl or piperidinyl group.

[0265] In some embodiments, the La is empty or selected from the following structures:

[0266] -C1-C 10 Alkylene-C(=O)- (e.g., -C2-C8 alkylene-C(=O)-, -C2-C7 alkylene-C(=O)-, -C2-C6 alkylene-C(=O)-), -C1-C 10 Alkylene-C(=O)NH- (e.g., -C2-C6 alkylene-C(=O)NH-, -C2-C5 alkylene-C(=O)NH-, -C2-C4 alkylene-C(=O)NH-),

[0267] -C1-C 10 Alkylene-C(=O)NH-(CH2CH2O) m -(e.g. -C2-C6 alkylene-C(=O)NH-(CH2CH2O) m -, -C2-C5 alkylene-C(=O)NH-(CH2CH2O) m -, -C2-C4 alkylene-C(=O)NH-(CH2CH2O) m -),

[0268] -C1-C 10 Alkylene-C(=O)NH-(CH2CH2O) m -C1-C 10 Alkylene-C(=O)-(e.g. -C2-C6 alkylene-C(=O)NH-(CH2CH2O) m-C2-C6 alkylene-C(=O)-, -C2-C5 alkylene-C(=O)NH-(CH2CH2O) m -C2-C5 alkylene-C(=O)-, -C2-C5 alkylene-C(=O)NH-(CH2CH2O) m -C2-C5 alkylene-C(=O)-),

[0269] m is any integer selected from 3 to 15, preferably 5-10, 6-9, 7-8.

[0270] In some embodiments, L1 is selected from the following structures:

[0271] -X 5 -C1-C 10 Alkylene-X 6 -、-X 5 -C1-C 10 Heteroalkylene-X 6 -、-X 5 -C3-C8 carbocyclyl-X 6 -、-X 5 -Arylene-X 6 -、-X 5 -heteroarylene-X 6 -、-X 5 -C1-C 10 Alkylene-arylene-X 6 -、-X 5 -C1-C 10 Alkylene-heteroarylene-X 6 -、-X 5 -Arylene-C1-C 10 Alkylene-X 6 -、-X 5 -heteroarylene-C1-C 10 Alkylene-X 6 -、-X 5 -C1-C 10 Alkylene-(C3-C8 carbocyclyl)-X 6 -、-X 5 -(C3-C8 carbocyclyl)-C1-C 10 Alkylene-X 6 -、-X 5 -C3-C8 heterocyclyl-X 6 -、-X 5 -C1-C 10 Alkylene-(C3-C8 heterocyclyl)-X 6 -、-X 5 -(C3-C8 heterocyclyl)-C1-C 10 Alkylene-X6 -、-X 5 -C3-C8 heterocyclyl-arylene-X 6 -、-X 5 -Arylene-C3-C8 heterocyclyl-X 6 -、-X 5 -C3-C8 heterocyclyl-heteroarylene-X 6 -、-X 5 -heteroarylene-C3-C8heterocyclyl-X 6 -;

[0272] wherein X 5 is absent or selected from O, S, NH, C(=O), NHC(=O), C(=O)NH, NHC(=O)NH, X 6 is absent or selected from O, S, NH, C(=O), NHC(=O), C(=O)NH, NHC(=O)NH;

[0273] Wherein, any methylene unit of L1 can be independently replaced by -O-, -S-, -N(R n )-、-CH(N(R n )2)-、-CH(P(R n )2)-、-C(N(R n )2)2-、-C(OR n )(N(R n )2)-, -C(=O)-, -OC(=O)-, -C(=O)O-, -N(R n )C(=O)-、-C(=O)N(R n )-、-SO-、-SO2-、-N(R n )SO2-、-SO2N(R n )-、-P(R n )-、-P(=O)(R n )-, -P(=O)(O)-, -C(=S)-, -C(=NR n )-、-SS-、-N=N-、-N(R n )-N=、=NN(R n )-, -C=N- or -N=C-; wherein each R n are independently selected from H, C1-C6 alkyl, C3-C8 carbocyclyl, C1-C6 haloalkyl, or two R n The groups combine with the nitrogen or phosphorus to which they are attached to form a 4-6 membered heterocycloalkyl ring, or an azetidinyl, pyrrolidinyl or piperidinyl group.

[0274] In some embodiments, L1 is empty or -C1-C10 Alkylene-C(=O)- (e.g., -C2-C8 alkylene-C(=O)-, -C2-C7 alkylene-C(=O)-, -C2-C6 alkylene-C(=O)-, -C2-C5 alkylene-C(=O)-, -C2-C4 alkylene-C(=O)-).

[0275] In some embodiments, the Cl-La-L1- is Preferably

[0276] In some embodiments, L2 is empty or selected from the following structures:

[0277] in Indicates the position of connection with the polypeptide, Indicates the location of the payload connection with a specific biological function;

[0278] m is any integer from 1 to 20;

[0279] n is any integer from 1 to 6;

[0280] R b Independently selected from H, C1-C6 alkyl, C3-C8 carbocyclyl, C1-C6 haloalkyl, C3-C8 heterocyclyl, C1-C6 alkyl-C3-C8 heterocyclyl, substituted aryl, substituted heteroaryl, C1-C6 alkyl-substituted aryl, C1-C6 alkyl-substituted heteroaryl.

[0281] Wherein any methylene unit of L2 can be independently replaced by -CHX-, -C(X2)-, -C3-C8 carbocyclyl-, -C3-C8 heterocyclyl-, -arylene-, -heteroarylene-, -O-, -S-, -N(R n )-、-CH(N(R n )2)-、-CH(P(R n )2)-、-C(N(R n )2)2-、-C(OR n )(N(R n )2)-, -C(=O)-, -OC(=O)-, -C(=O)O-, -N(R n )C(=O)-、-C(=O)N(R n )-、-SO-、-SO2-、-N(R n )SO2-、-SO2N(R n )-、-P(R n )-、-P(=O)(Rn )-, -P(=O)(O)-, -C(=S)-, -C(=NR n )-、-SS-、-N=N-、-N(R n )-N=、=NN(R n )-, -C=N- or -N=C-; wherein X represents halogen or deuterium, each R n are independently selected from H, C1-C6 alkyl, C3-C8 carbocyclyl, C1-C6 haloalkyl, or two R n The groups combine with the nitrogen or phosphorus to which they are attached to form a 4-6 membered heterocycloalkyl ring, or an azetidinyl, pyrrolidinyl or piperidinyl group.

[0282] In some embodiments, L2 is empty or selected from the following structures:

[0283] in Indicates the position of connection with the polypeptide, Indicates the location of the payload connection with a specific biological function;

[0284] W and Y are each independently N or CR Y , and each R Y are independently H or C1-C 10 alkyl;

[0285] m is any integer from 0 to 20;

[0286] X is halogen or deuterium.

[0287] In some embodiments, L2 is empty or selected from:

[0288] (For example ),

[0289] (For example ),

[0290] (For example ),

[0291] (For example ),

[0292] n is any integer selected from 1 to 6, 1-5, 1-4 or 1-3.

[0293] In some embodiments, L2 is empty or selected from -PAB- -PAB-DMEDA- -NMEDA-

[0294] In some embodiments, the linker is selected from the following structures:

[0295] -(Succinimidyl-3-yl-N)-CH2-C(=O)-P1-P2-;

[0296] -(Succinimidyl-3-yl-N)-CH2CH2-C(=O)-P1-P2-;

[0297] -(succinimidyl-3-yl-N)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-;

[0298] -(Succinimidyl-3-yl-N)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-;

[0299] -(Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-;

[0300] -(Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-;

[0301] -(Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-;

[0302] -(Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-;

[0303] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-;

[0304] -(Succinimidyl-3-yl-N)-CH2-C(=O)-P1-P2-PAB-;

[0305] -(Succinimidyl-3-yl-N)-CH2CH2-C(=O)-P1-P2-PAB-;

[0306] -(Succinimidyl-3-yl-N)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-;

[0307] -(Succinimidyl-3-yl-N)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-;

[0308] -(Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-;

[0309] -(Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-;

[0310] -(Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-;

[0311] -(Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-;

[0312] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-PAB-;

[0313] in:

[0314] Indicates the connection position with the biological ligand unit

[0315] PAB represents the following structure:

[0316] The -P1-P2- represents the following structure:

[0317] -His-Val-Leu-Asn-Leu-Val-Cit-(SEQ ID NO: 1);

[0318] -Val-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:2);

[0319] -Ile-Pro-Val-Ser-Leu-Val-Cit-(SEQ ID NO:3);

[0320] -Gly-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:4);

[0321] -Gly-Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:5);

[0322] -Gly-Pro-Ala-Gly-Leu-Val-Cit-(SEQ ID NO:6);

[0323] -Gly-Pro-Gln-Gly-Leu-Val-Cit-(SEQ ID NO:7);

[0324] -Gly-Pro-Leu-Gly-Leu-Val-Cit-(SEQ ID NO:8);

[0325] -Gly-Pro-Ser-Gly-Leu-Val-Cit-(SEQ ID NO:9);

[0326] -Gly-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:10);

[0327] -Gly-Pro-Ala-Ala-Leu-Val-Cit-(SEQ ID NO:11);

[0328] -Gly-Pro-Ser-Ala-Leu-Val-Cit-(SEQ ID NO:12);

[0329] -His-Val-Leu-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:13);

[0330] -Val-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:14);

[0331] -Ile-Pro-Val-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:15);

[0332] -Gly-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:16);

[0333] -Gly-Pro-Ala-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:17);

[0334] -Gly-Pro-Ala-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:18);

[0335] -Gly-Pro-Gln-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:19);

[0336] -Gly-Pro-Leu-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:20);

[0337] -Gly-Pro-Ser-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:21);

[0338] -Gly-Pro-Arg-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:22);

[0339] -Gly-Pro-Ala-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:23);

[0340] -Gly-Pro-Ser-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:24);

[0341] -Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:25);

[0342] -Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:26);

[0343] -Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:27);

[0344] -Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:28);

[0345] -Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:29);

[0346] -Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:30);

[0347] -Gly-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:31);

[0348] -Gly-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:32);

[0349] -Gly-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:33);

[0350] -Gly-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:34);

[0351] -Gly-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:35);

[0352] -His-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:36);

[0353] -His-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:37);

[0354] -His-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:38);

[0355] -His-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:39);

[0356] -His-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:40);

[0357] -His-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:41);

[0358] -Ser-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:42);

[0359] -Ser-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:43);

[0360] -Ser-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:44);

[0361] -Ser-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:45);

[0362] -Ser-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:46);

[0363] -Ser-Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:47);

[0364] -Ser-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:48);

[0365] -Gly-Pro-Ala-Pro-Leu-Val-Cit-(SEQ ID NO:49);

[0366] -Gly-Pro-Ala-Asn-Leu-Val-Cit-(SEQ ID NO:50);

[0367] -Gly-Pro-Arg-Gly-Dap(Me2)-Val-Cit-(SEQ ID NO:51);

[0368] -Gly-Pro-Arg-Asn-Dap(Me2)-Val-Cit-(SEQ ID NO:60);

[0369] -Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:61);

[0370] -Gly-Pro-Arg-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:62);

[0371] -Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:63);

[0372] -Gly-Pro-Cit-Asn-Leu-Val-Cit-(SEQ ID NO:64);

[0373] -Ser-Pro-Cit-Ser-Dap(Me2)-Val-Cit-(SEQ ID NO:65);

[0374] -Pro-Arg-Ser-Dap(Me2)-Val-Cit-(SEQ ID NO:66);

[0375] -Pro-Cit-Asn-Leu-Val-Cit-(SEQ ID NO:67);

[0376] -Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:68);

[0377] Or a variant having one or two mismatches relative to the sequence of any one of SEQ ID NOs: 1-51 and 60-68.

[0378] In some embodiments, the linker is selected from:

[0379] -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)-P1-P2-;

[0380] -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)NH-P1-P2-;

[0381] -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)NH-(CH2CH2O) m -P1-P2-;

[0382] -(Succinimide-3-yl-N)-C1-C10alkylene-C(=O)NH-(CH2CH2O) m -C1-C 10 Alkylene-C(=O)-P1-P2-;

[0383] -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)-P1-P2-PAB-;

[0384] -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)NH-P1-P2-PAB-;

[0385] -(Succinimide-3-yl-N)-C1-C10alkylene-C(=O)NH-(CH2CH2O) m -P1-P2-PAB-;

[0386] -(Succinimide-3-yl-N)-C1-C10alkylene-C(=O)NH-(CH2CH2O) m -C1-C 10 Alkylene-C(=O)-P1-P2-PAB-;

[0387] -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)-P1-P2-PAB-DMEDA-;

[0388] -(Succinimidyl-3-yl-N)-C1-C10 alkylene-C(=O)NH-P1-P2-PAB-DMEDA-;

[0389] -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)NH-(CH2CH2O)m -P1-P2-PAB-DMEDA-;

[0390] -(Succinimide-3-yl-N)-C1-C10alkylene-C(=O)NH-(CH2CH2O) m -C1-C 10 Alkylene-C(=O)-P1-P2-PAB-DMEDA-;

[0391] -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)-P1-P2-NMEDA-;

[0392] -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)NH-P1-P2-NMEDA-;

[0393] -(Succinimide-3-yl-N)-C1-C10alkylene-C(=O)NH-(CH2CH2O) m -P1-P2-NMEDA-;

[0394] -(Succinimide-3-yl-N)-C1-C10alkylene-C(=O)NH-(CH2CH2O) m -C1-C 10 Alkylene-C(=O)-P1-P2-NMEDA-;

[0395] -(C=O)-C1-C10 alkylene-C(=O)-P1-P2-;

[0396] -(C=O)-C1-C10 alkylene-C(=O)-P1-P2-PAB-;

[0397] -(C=O)-C1-C10 alkylene-C(=O)NH-P1-P2-PAB-;

[0398] -(C=O)-C1-C10 alkylene-C(=O)-P1-P2-PAB-DMEDA-;

[0399] -(C=O)-C1-C10 alkylene-C(=O)NH-P1-P2-PAB-DMEDA-;

[0400] -(C=O)-C1-C10 alkylene-C(=O)-P1-P2-NMEDA-;

[0401] -(C=O)-C1-C10 alkylene-C(=O)NH-P1-P2-NMEDA-;

[0402] m is any integer selected from 3-15, preferably 5-10, 6-9, 7-8,

[0403] n is any integer selected from 1-10, preferably 2-6, 2-5 or 2-4.

[0404] In some embodiments, the linker is selected from the following structures:

[0405] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-P1-P2-;

[0406] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-P1-P2-PAB-;

[0407] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-P1-P2-PAB-DMEDA-;

[0408] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-P1-P2-NMEDA-;

[0409] -(C=O)-C6alkylene-C(=O)-P1-P2-;

[0410] -(C=O)-C6alkylene-C(=O)-P1-P2-PAB-;

[0411] -(C=O)-C6alkylene-C(=O)-P1-P2-PAB-DMEDA-;

[0412] -(C=O)-C6alkylene-C(=O)-P1-P2-NMEDA-;

[0413] -(Succinimide-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-P1-P2-;

[0414] -(Succinimide-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-P1-P2-PAB-;

[0415] -(Succinimide-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-P1-P2-PAB-DMEDA-;

[0416] -(Succinimide-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-P1-P2-NMEDA-;

[0417] In some embodiments, the linker is one or more selected from the following:

[0418] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAGLVCit-PAB-;

[0419] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPLGLGGFG-;

[0420] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPASLVCit-PAB-;

[0421] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRGLVCit-PAB-;

[0422] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPSALVCit-PAB-;

[0423] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRGLGGFG-;

[0424] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAALGGFG-;

[0425] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRNLVCit-PAB-;

[0426] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLVCit-PAB-;

[0427] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNDap(Me2)VCit-PAB-;

[0428] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRNLVCit-PAB-;

[0429] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPLSLVCit-PAB-;

[0430] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRGLVCit-PAB-;

[0431] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRPLVCit-PAB-;

[0432] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRSLVCit-PAB-;

[0433] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRPLVCit-PAB-;

[0434] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPCitPLVCit-PAB-;

[0435] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-HPRGLVCit-PAB-;

[0436] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-HPCitPLVCit-PAB-;

[0437] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitGLVCit-PAB-;

[0438] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAPLVCit-PAB-;

[0439] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PLSLVCit-PAB-;

[0440] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitSLVCit-PAB-;

[0441] -(C=O)-C6alkylene-C(=O)-GPRNLVCit-PAB-;

[0442] -(Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-GPRNLVCit-PAB-;

[0443] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLVCit-PAB-DMEDA-;

[0444] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLGGFG-;

[0445] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLGGFG-NMEDA-;

[0446] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRSLVCit-PAB-;

[0447] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRSLVCit-PAB-;

[0448] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PCitSLVCit-PAB-;

[0449] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PCitPLVCit-PAB-;

[0450] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PCitGLVCit-PAB-;

[0451] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPCitGLVCit-PAB-;

[0452] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRGLVCit-PAB-;

[0453] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRPLVCit-PAB-;

[0454] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitPLVCit-PAB-;

[0455] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPANLVCit-PAB-;

[0456] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPCitNLVCit-PAB-;

[0457] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitSDap(Me2)VCit-PAB-;

[0458] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRSDap(Me2)VCit-PAB-;

[0459] -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PCitNLVCit-PAB-; and

[0460] -(Succinimid-3-yl-N)-C5alkylene-C(=O)-PASLVCit-PAB-.

[0461] In some embodiments, the linker is selected from the following structures:

[0462] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLGLGGFG-;

[0463] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLGGFG-;

[0464] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAALGGFG-;

[0465] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-;

[0466] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPASLVCit-PAB-;

[0467] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLVCit-PAB-;

[0468] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPSALVCit-PAB-;

[0469] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-PRNLVCit-PAB-;

[0470] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRNLVCit-PAB-;

[0471] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGDap(Me2)VCit-PAB-;

[0472] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-SPRNLVCit-PAB-;

[0473] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLSLVCit-PAB-;

[0474] Wherein, PAB represents the following structure:

[0475] Dap(Me2) represents the following structure:

[0476] On the other hand, the present application provides a linker drug (also called a "linker-payload intermediate"), which comprises the above-mentioned polypeptide or polypeptide fragment having the structure shown by -P1-P2-.

[0477] In some embodiments, the linker drug has the following structure:

[0478] C1'—La—L1—P1—P2—L2-D,

[0479] Wherein, the C1' is a functional group that reacts with the biological ligand unit;

[0480] La is empty or a tuning spacer;

[0481] L1 is empty or a spacer;

[0482] P1-P2 are the polypeptides or polypeptide fragments described in any one of the embodiments herein;

[0483] L2 is empty or a spacer, used to connect a payload with a specific biological function;

[0484] D is the drug unit;

[0485] The positions of La and L1 can be swapped.

[0486] In some embodiments, the coordination spacer linker La is the structural unit described in the above aspect.

[0487] In some embodiments, the spacer L1 is the spacer described in the previous aspect.

[0488] In some embodiments, the spacer L2 is the spacer described in the previous aspect.

[0489] In some embodiments, C1' is selected from a functional group that reacts with an amino group, a sulfhydryl group, a keto group, an azide group, or an alkyne group.

[0490] In some embodiments, C1' is selected from an enzyme-catalyzed reaction substrate.

[0491] In some embodiments, C1' is selected from the following structures:

[0492] in:

[0493] R X Selected from halogen, -SPh, R S Selected from halogen, sulfone, tertiary amine salt, diazonium salt, -OMs, MeSO2-, and CF3SO3-;

[0494] Indicates the connection position with La.

[0495] In some embodiments, C1' is selected from the following structures:

[0496] In some embodiments, the C1'-La-L1- is MC-, MeO2S-Pym-, NHS- or Mal-PEG8- are preferred.

[0497] In some embodiments, the drug unit D is selected from an immunomodulator, a protein degrader, and a cytotoxic agent.

[0498] In some embodiments, the Drug unit D is selected from a cytotoxic agent.

[0499] In some embodiments, the drug unit comprises: amanitins, anthracyclines, auristatins, baccatins, calicheamicins, camptothecins, cemadotins, colchicines, colcimids, combretastatins, cryptophycins, discodermolides, duocarmycins, docetaxel, doxorubicin, duocarmycins, echinomycins, eleutherobins, epothilones, estramustines, lexitropsins, maytansines, maytansinoids, methotrexate, netropsins, pyrrolo[2,1-c][1,4]benzodi-azepines (PBDs), puromycins, rhizoxins, SN-38, taxanes, tubulysins, vincaalkaloids, tetrahydroisoquinoline alkaloids or derivatives thereof, protein degraders or derivatives thereof.

[0500] In some embodiments, the drug unit comprises a DNA topoisomerase I inhibitor, a tubulin inhibitor, a DNA replication inhibitor, or a protein degrader.

[0501] In some embodiments, the drug unit comprises maytansine alkaloid or its derivatives, camptothecin or its derivatives, auristatin or its derivatives, tetrahydroisoquinoline alkaloid or its derivatives, BTK protein degrader and its derivatives, GSPT1 protein degrader and its derivatives.

[0502] In some embodiments, the drug unit comprises DM1, DM4, DX8951, MMAE, Dxd, SN38, Trabectedin (ET743), Lurbinectedin, CC885, CC-90009.

[0503] In some embodiments, the linker drug is selected from:

[0504] MC-P1-P2-D;

[0505] MC-P1-P2-PAB-D;

[0506] MC-P1-P2-PAB-DMEDA-D;

[0507] MC-P1-P2-NMEDA-D;

[0508] MeO2S-Pym-P1-P2-D;

[0509] MeO2S-Pym-P1-P2-PAB-D;

[0510] MeO2S-Pym-P1-P2-PAB-DMEDA-D;

[0511] MeO2S-Pym-P1-P2-NMEDA-D;

[0512] NHS-P1-P2-D;

[0513] NHS-P1-P2-PAB-D;

[0514] NHS-P1-P2-PAB-DMEDA-D;

[0515] NHS-P1-P2-NMEDA-D;

[0516] Mal-PEG8-P1-P2-D;

[0517] Mal-PEG8-P1-P2-PAB-D;

[0518] Mal-PEG8-P1-P2-PAB-DMEDA-D;

[0519] Mal-PEG8-P1-P2-NMEDA-D;

[0520] DBCO-P1-P2-D;

[0521] DBCO-P1-P2-PAB-D;

[0522] DBCO-P1-P2-PAB-DMEDA-D;

[0523] DBCO-P1-P2-NMEDA-D;

[0524] Hydrazide-P1-P2-D;

[0525] Hydrazide-P1-P2-PAB-D;

[0526] Hydrazide-P1-P2-PAB-DMEDA-D;

[0527] Hydrazide-P1-P2-NMEDA-D.

[0528] In some preferred embodiments, the drug unit D comprises DX8951, MMAE, Dxd, SN38 or Trabectedin (ET743).

[0529] In some embodiments, the linker drug has the following structure:

[0530] (Maleimido-N-yl)-CH2-C(=O)-P1-P2-D;

[0531] (Maleimido-N-yl)-CH2CH2-C(=O)-P1-P2-D;

[0532] (Maleimido-N-yl)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D;

[0533] (Maleimide-N-yl)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D;

[0534] (Maleimido-N-yl)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D;

[0535] (Maleimido-N-yl)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D;

[0536] (Maleimido-N-yl)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D;

[0537] (Maleimido-N-yl)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D;

[0538] (Maleimido-N-yl)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-D;

[0539] (Maleimido-N-yl)-CH2-C(=O)-P1-P2-PAB-D;

[0540] (Maleimido-N-yl)-CH2CH2-C(=O)-P1-P2-PAB-D;

[0541] (Maleimido-N-yl)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D;

[0542] (Maleimide-N-yl)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D;

[0543] (Maleimido-N-yl)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D;

[0544] (Maleimido-N-yl)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D;

[0545] (Maleimido-N-yl)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D;

[0546] (Maleimido-N-yl)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D;

[0547] (Maleimido-N-yl)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-PAB-D;

[0548] (N-hydroxysuccinimide ester-1-yl)-CH2-C(=O)-P1-P2-D;

[0549] (N-hydroxysuccinimide ester-1-yl)-CH2CH2-C(=O)-P1-P2-D;

[0550] (N-hydroxysuccinimide ester-1-yl)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D;

[0551] (N-hydroxysuccinimide ester-1-yl)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D;

[0552] (N-hydroxysuccinimide ester-1-yl)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D;

[0553] (N-hydroxysuccinimide ester-1-yl)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D;

[0554] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2-C(=O)-P1-P2-D;

[0555] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D;

[0556] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D;

[0557] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2C(=O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D;

[0558] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2C(=O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D;

[0559] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2O-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D;

[0560] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2O-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D;

[0561] (N-hydroxysuccinimide ester-1-yl)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-D;

[0562] (N-hydroxysuccinimide ester-1-yl)-CH2-C(=O)-P1-P2-PAB-D;

[0563] (N-hydroxysuccinimide ester-1-yl)-CH2CH2-C(=O)-P1-P2-PAB-D;

[0564] (N-hydroxysuccinimide ester-1-yl)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D;

[0565] (N-hydroxysuccinimide ester-1-yl)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D;

[0566] (N-hydroxysuccinimide ester-1-yl)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D;

[0567] (N-hydroxysuccinimide ester-1-yl)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D;

[0568] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2-C(=O)-P1-P2-PAB-D;

[0569] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D;

[0570] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D;

[0571] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2C(=O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D;

[0572] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2C(=O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D;

[0573] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2O-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D;

[0574] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2O-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D;

[0575] (N-hydroxysuccinimide ester-1-yl)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-PAB-D;

[0576] Gly-Gly-Gly-NH-CH2CH2CH2CH2CH2-C(=O)-P1-P2-D;

[0577] Gly-Gly-Gly-NH-CH2CH2CH2CH2CH2-C(=O)-P1-P2-PAB-D;

[0578] The (maleimide-N-yl)- has the following structure:

[0579] The (N-hydroxysuccinimide ester-1-yl)- has the following structure:

[0580] in Indicates the connection position with La, L1 or P1-P2.

[0581] The -P1-P2- represents the following structure:

[0582] -His-Val-Leu-Asn-Leu-Val-Cit-(SEQ ID NO: 1);

[0583] -Val-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:2);

[0584] -Ile-Pro-Val-Ser-Leu-Val-Cit-(SEQ ID NO:3);

[0585] -Gly-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:4);

[0586] -Gly-Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:5);

[0587] -Gly-Pro-Ala-Gly-Leu-Val-Cit-(SEQ ID NO: 6);

[0588] -Gly-Pro-Gln-Gly-Leu-Val-Cit-(SEQ ID NO:7);

[0589] -Gly-Pro-Leu-Gly-Leu-Val-Cit-(SEQ ID NO:8);

[0590] -Gly-Pro-Ser-Gly-Leu-Val-Cit-(SEQ ID NO:9);

[0591] -Gly-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:10);

[0592] -Gly-Pro-Ala-Ala-Leu-Val-Cit-(SEQ ID NO:11);

[0593] -Gly-Pro-Ser-Ala-Leu-Val-Cit-(SEQ ID NO:12);

[0594] -His-Val-Leu-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:13);

[0595] -Val-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:14);

[0596] -Ile-Pro-Val-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:15);

[0597] -Gly-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:16);

[0598] -Gly-Pro-Ala-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:17);

[0599] -Gly-Pro-Ala-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:18);

[0600] -Gly-Pro-Gln-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:19);

[0601] -Gly-Pro-Leu-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:20);

[0602] -Gly-Pro-Ser-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:21);

[0603] -Gly-Pro-Arg-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:22);

[0604] -Gly-Pro-Ala-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:23);

[0605] -Gly-Pro-Ser-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:24);

[0606] -Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:25);

[0607] -Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:26);

[0608] -Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:27);

[0609] -Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:28);

[0610] -Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:29);

[0611] -Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:30);

[0612] -Gly-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:31);

[0613] -Gly-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:32);

[0614] -Gly-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:33);

[0615] -Gly-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:34);

[0616] -Gly-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:35);

[0617] -His-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:36);

[0618] -His-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:37);

[0619] -His-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:38);

[0620] -His-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:39);

[0621] -His-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:40);

[0622] -His-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:41);

[0623] -Ser-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:42);

[0624] -Ser-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:43);

[0625] -Ser-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:44);

[0626] -Ser-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:45);

[0627] -Ser-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:46);

[0628] -Ser-Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:47);

[0629] -Ser-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:48);

[0630] -Gly-Pro-Ala-Pro-Leu-Val-Cit-(SEQ ID NO:49);

[0631] -Gly-Pro-Ala-Asn-Leu-Val-Cit-(SEQ ID NO:50);

[0632] -Gly-Pro-Arg-Gly-Dap(Me2)-Val-Cit-(SEQ ID NO:51);

[0633] -Gly-Pro-Arg-Asn-Dap(Me2)-Val-Cit-(SEQ ID NO:60);

[0634] -Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:61);

[0635] -Gly-Pro-Arg-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO: 62);

[0636] -Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:63);

[0637] -Gly-Pro-Cit-Asn-Leu-Val-Cit-(SEQ ID NO: 64);

[0638] -Ser-Pro-Cit-Ser-Dap(Me2)-Val-Cit-(SEQ ID NO:65);

[0639] -Pro-Arg-Ser-Dap(Me2)-Val-Cit-(SEQ ID NO:66);

[0640] -Pro-Cit-Asn-Leu-Val-Cit-(SEQ ID NO:67);

[0641] -Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:68);

[0642] Or a variant having one or two mismatches relative to the sequence of any one of SEQ ID NOs: 1-51 and 60-68.

[0643] The D is camptothecin or its derivatives, auristatin or its derivatives, tetrahydroisoquinoline alkaloids or its derivatives, PROTAC protein degraders, molecular glue protein degraders, preferably DX8951, MMAE, Dxd, SN38, ET743, CC885, CC-90009.

[0644] In some embodiments, the linker drug is one or more selected from the following:

[0645] MC-GPAGLVCit-PAB-DX8951;

[0646] MC-GPAGLVCit-PAB-MMAE;

[0647] MC-GPAGLVCit-PAB-ET743;

[0648] MC-GPLGLGGFG-DX8951;

[0649] MC-GPASLVCit-PAB-DX8951;

[0650] MC-GPRGLVCit-PAB-DX8951;

[0651] MC-GPRGLVCit-PAB-MMAE;

[0652] MC-GPSALVCit-PAB-DX8951;

[0653] MC-GPRGLGGFG-DX8951;

[0654] MC-GPAALGGFG-DX8951;

[0655] MC-PRNLVCit-PAB-DX8951;

[0656] MC-PRNLVCit-PAB-MMAE;

[0657] MC-GPRNLVCit-PAB-DX8951;

[0658] MC-GPRNLVCit-PAB-MMAE;

[0659] MC-GPRNLVCit-PAB-ET743;

[0660] MC-GPRNDap(Me2)VCit-PAB-DX8951;

[0661] MC-GPRNDap(Me2)VCit-PAB-MMAE;

[0662] MC-SPRNLVCit-PAB-DX8951;

[0663] MC-SPRNLVCit-PAB-MMAE;

[0664] MC-GPLSLVCit-PAB-DX8951;

[0665] MC-GPLSLVCit-PAB-MMAE;

[0666] MC-PRGLVCit-PAB-DX8951;

[0667] MC-PRPLVCit-PAB-DX8951;

[0668] MC-GPRSLVCit-PAB-DX8951;

[0669] MC-GPRPLVCit-PAB-DX8951;

[0670] MC-GPCitPLVCit-PAB-DX8951;

[0671] MC-HPRGLVCit-PAB-DX8951;

[0672] MC-HPCitPLVCit-PAB-DX8951;

[0673] MC-SPCitGLVCit-PAB-DX8951;

[0674] MC-GPAPLVCit-PAB-DX8951;

[0675] MC-PLSLVCit-PAB-DX8951;

[0676] MC-SPCitSLVCit-PAB-DX8951;

[0677] MC-SPCitSLVCit-PAB-MMAE;

[0678] MC-SPCitSLVCit-PAB-ET743;

[0679] MeO2S-Pym-GPRNLVCit-PAB-DX8951;

[0680] DBCO-GPRNLVCit-PAB-DX8951;

[0681] NHS-GPRNLVCit-PAB-DX8951;

[0682] Hydrazide-GPRNLVCit-PAB-DX8951;

[0683] Mal-PEG8-GPRNLVCit-PAB-DX8951;

[0684] MC-GPRNLVCit-PAB-DMEDA-SN38;

[0685] MC-GPRNLGGFG-Dxd;

[0686] MC-GPRNLGGFG-NMEDA-Dxd;

[0687] MC-PRSLVCit-PAB-DX8951;

[0688] MC-SPRSLVCit-PAB-DX8951

[0689] MC-PCitSLVCit-PAB-DX8951;

[0690] MC-PCitPLVCit-PAB-DX8951;

[0691] MC-PCitGLVCit-PAB-DX8951;

[0692] MC-GPCitGLVCit-PAB-DX8951;

[0693] MC-SPRGLVCit-PAB-DX8951;

[0694] MC-SPRPLVCit-PAB-DX8951;

[0695] MC-SPCitPLVCit-PAB-DX8951;

[0696] MC-GPANLVCit-PAB-DX8951;

[0697] MC-GPCitNLVCit-PAB-DX8951;

[0698] MC-SPCitSDap(Me2)VCit-PAB-DX8951;

[0699] MC-SPCitSDap(Me2)VCit-PAB-MMAE;

[0700] MC-PRSDap(Me2)VCit-PAB-DX8951;

[0701] MC-PRSDap(Me2)VCit-PAB-MMAE;

[0702] MC-PCitNLVCit-PAB-DX8951; and

[0703] MC-PASLVCit-PAB-DX8951.

[0704] In some embodiments, the linker drug has the following structure:

[0705] MC-GPAGLVCit-PAB-DX8951;

[0706] MC-GPLGLGGFG-DX8951;

[0707] MC-GPASLVCit-PAB-DX8951;

[0708] MC-GPRGLVCit-PAB-DX8951;

[0709] MC-GPSALVCit-PAB-DX8951;

[0710] MC-GPRGLGGFG-DX8951;

[0711] MC-GPAALGGFG-DX8951;

[0712] MC-PRNLVCit-PAB-DX8951;

[0713] MC-GPRNLVCit-PAB-DX8951;

[0714] MC-GPRGDap(Me2)LVCit-PAB-DX8951;

[0715] MC-SPRNLVCit-PAB-DX8951;

[0716] MC-GPLSLVCit-PAB-DX8951;

[0717] MC-PRGLVCit-PAB-DX8951;

[0718] MC-PRPLVCit-PAB-DX8951;

[0719] MC-GPRSLVCit-PAB-DX8951;

[0720] MC-GPRPLVCit-PAB-DX8951;

[0721] MC-GPCitPLVCit-PAB-DX8951;

[0722] MC-HPRGLVCit-PAB-DX8951;

[0723] MC-HPCitPLVCit-PAB-DX8951;

[0724] MC-SPCitGLVCit-PAB-DX8951;

[0725] MC-GPAPLVCit-PAB-DX8951;

[0726] MC-PLSLVCit-PAB-DX8951;

[0727] MC-SPCitSLVCit-PAB-DX8951;

[0728] MC-PRSLVCit-PAB-DX8951;

[0729] MC-SPRSLVCit-PAB-DX8951;

[0730] MC-PCitSLVCit-PAB-DX8951;

[0731] MC-PCitPLVCit-PAB-DX8951;

[0732] MC-PCitGLVCit-PAB-DX8951;

[0733] MC-GPCitGLVCit-PAB-DX8951;

[0734] MC-SPRGLVCit-PAB-DX8951;

[0735] MC-SPRPLVCit-PAB-DX8951;

[0736] MC-SPCitPLVCit-PAB-DX8951;

[0737] MC-GPANLVCit-PAB-DX8951

[0738] MC-GPCitNLVCit-PAB-DX8951;

[0739] MC-SPCitSDap(Me2)VCit-PAB-DX8951;

[0740] MC-PRSDap(Me2)VCit-PAB-DX8951;

[0741] MC-GPAGLVCit-PAB-MMAE;

[0742] MC-GPRGLVCit-PAB-MMAE;

[0743] MC-PRNLVCit-PAB-MMAE;

[0744] MC-GPRNLVCit-PAB-MMAE;

[0745] MC-GPRGDap(Me2)LVCit-PAB-MMAE;

[0746] MC-SPRNLVCit-PAB-MMAE;

[0747] MC-GPLSLVCit-PAB-MMAE;

[0748] MC-SPCitSLVCit-PAB-MMAE

[0749] MC-SPCitSDap(Me2)VCit-PAB-MMAE

[0750] MC-SPCitSDap(Me2)VCit-PAB-MMAE

[0751] MC-GPAGLVCit-PAB-ET743;

[0752] MC-GPRNLVCit-PAB-ET743;

[0753] MC-SPCitSLVCit-PAB-ET743;

[0754] The MC- has the following structure:

[0755] in Indicates the position of connection with the polypeptide,

[0756] The DX8951 has the following structure:

[0757] in Indicates the connection position between DX8951 and the linker,

[0758] The MMAE has the following structure:

[0759] in Indicates the connection position between MMAE and the linker,

[0760] The ET-743 has the following structure:

[0761] in Indicates the connection position between ET743 and the linker.

[0762] On the other hand, the present application provides a bioligand-drug conjugate, such as an antibody-drug conjugate, wherein the conjugate comprises the above-mentioned polypeptide or polypeptide fragment having the structure represented by -P1-P2-.

[0763] In some embodiments, the conjugate has the general formula:

[0764] in:

[0765] Bp is the biological ligand;

[0766] L is a linker comprising a polypeptide or polypeptide fragment having a structure represented by -P1-P2-;

[0767] D is the drug unit;

[0768] p is any value between 1 and 20 (eg, any integer).

[0769] In some embodiments, the conjugate has the general formula:

[0770] in:

[0771] Bp is the biological ligand;

[0772] C1, La, L1, L2 are defined as any one of the embodiments herein;

[0773] The -P1-P2- unit is a polypeptide or polypeptide fragment as described in some embodiments herein;

[0774] D is the drug unit;

[0775] p is any value between 1 and 20 (eg, any integer).

[0776] In some embodiments, the drug unit D is defined as described in any one of the above aspects.

[0777] In some embodiments, the biological ligand targets a cell surface receptor or a tumor associated antigen.

[0778] In some embodiments, the cell surface receptors or tumor-associated antigens include: EGFR, HER2, HER3, c-Met, claudin18.2, TROP-2, folate receptor alpha (FRα), ROR1, ROR2, BCMA, PSMA, CD19, CD20, CD22, CD30, CD33, CD37, CD46, CD48, CD56, CD79b, CD123, CD138, CD166, CD276, CEACAM5, SDC1, CD74, CD70, MUC1, MUC16, GUCY2C, MSLN, SCL34A2, FOLH1, TPBC, PVRL4, STEAP1, SCL44A4, NACM1, EDNRB, GPNMB, FGFR, SEZ6, Nectin-4, PD-L1, Tissue Factor(TF), B7-H3, B7-H4, LY6E, LIV-1, CDH4, CDH6, ITGB6, GPC1, GPC3, ENPP3, KAAG1, FZD7, SSTR2, DLK1, T MEFF1, TM4SF1, SLAMF7, DLL3, FLT3, TNFRSF10B, EPCAM, AXL, IL2RA, 5T4, FAP, ICAM-1, IGF-1R, TNFR1, GLP-1.

[0779] In some embodiments, the bioligand unit includes, but is not limited to, an antibody, an antibody fragment, a protein, a polypeptide, a polynucleotide, or a chemical small molecule. In some embodiments, the bioligand includes, but is not limited to, an antibody or an antigen-binding fragment thereof, a modified antibody or an antigen-binding fragment thereof, an oligonucleotide, a DNA fragment, an aptamer, a polypeptide, a nanoparticle with targeting properties, or a chemically synthesized small molecule.

[0780] In some embodiments, the biological ligand comprises an antibody or antigen-binding fragment thereof, or a modified antibody or antigen-binding fragment thereof.

[0781] In some embodiments, the antibody comprises a monoclonal antibody, a polyclonal antibody, a dimer, a multimer, a multispecific antibody, a whole antibody, an antibody fragment, a human antibody, a humanized antibody, or a chimeric antibody.

[0782] In some embodiments, the modified antibodies include modified monoclonal antibodies, modified polyclonal antibodies, modified dimers, modified multimers, modified multispecific antibodies, modified whole antibodies, modified antibody fragments, modified human antibodies, modified humanized antibodies, or modified chimeric antibodies.

[0783] In some embodiments, the antigen binding fragment comprises a Fab, a Fab', an Fv fragment, a F(ab')2, a scFv, a di-scFv and / or a dAb.

[0784] In some embodiments, the modified antigen-binding fragment comprises a modified Fab, a modified Fab', a modified Fv fragment, a modified F(ab')2, a modified scFv, a modified di-scFv and / or a modified dAb.

[0785] In some embodiments, the antibody is a monoclonal antibody or a modified monoclonal antibody.

[0786] In some embodiments, the antibody is a human antibody, a humanized antibody, or a chimeric antibody, wherein the modified antibody is a modified human antibody, a modified humanized antibody, or a modified chimeric antibody.

[0787] In some embodiments, the modified methods are: amino acid point mutation, glycosylation modification, and polypeptide modification.

[0788] In some embodiments, the antibody or antigen-binding fragment thereof, modified antibody or modified antigen-binding fragment thereof is: anti-EGFR, anti-HER2, anti-HER3, anti-c-Met, anti-claudin18.2, anti-TROP-2, anti-FRα, anti-ROR1, anti-ROR2, anti-BCMA, anti-PSMA, anti-CD19, anti-CD20, anti-CD22, anti-CD30, anti-CD33, anti- CD37, anti-CD46, anti-CD48, anti-CD56, anti-CD79b, anti-CD123, anti-CD138, anti-CD166, anti-CD276, anti-CEACAM5, anti -SDC1, anti-CD74, anti-CD70, anti-MUC1, anti-MUC16, anti-GUCY2C, anti-MSLN, anti-SCL34A2, anti-FOLH1, anti-TPBC, anti- PVRL4, anti-STEAP1, anti-SCL44A4, anti-NACM1, anti-EDNRB, anti-GPNMB, anti-FGFR, anti-SEZ6, anti-Nectin-4, anti-PD-L 1. anti-TF, anti-B7-H3, anti-B7-H4, anti-LY6E, anti-LIV-1, anti-CDH4, anti-CDH6, anti-ITGB6, anti-GPC1, anti-GPC3, anti -ENPP3, anti-KAAG1, anti-FZD7, anti-SSTR2, anti-DLK1, anti-TMEFF1, anti-TM4SF1, anti-SLAMF7, anti-DLL3, anti-FLT3, an ti-TNFRSF10B, anti-EPCAM, anti-AXL, anti-IL2RA, anti-5T4, anti-FAP, anti-ICAM-1, anti-IGF-1R, anti-TNFR1, anti-GLP-1.

[0789] In some embodiments, the antibody or antigen-binding fragment thereof, modified antibody or modified antigen-binding fragment thereof is: anti-Her2, anti-TF, anti-ROR1, anti-GPC3 antibody or antigenic structural fragment thereof.

[0790] In some embodiments, the antibody is: Trastuzumab, Tisotumab, Zilovertamab, Codrituzumab.

[0791] In some embodiments, the antibody drug conjugate is selected from the following structures:

[0792] Ab-((Succinimidyl-3-yl-N)-CH2-C(=O)-P1-P2-D) n ;

[0793] Ab-((Succinimidyl-3-yl-N)-CH2CH2-C(=O)-P1-P2-D) n ;

[0794] Ab-((Succinimidyl-3-yl-N)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D) n ;

[0795] Ab-((Succinimidyl-3-yl-N)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D) n ;

[0796] Ab-((Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D) n ;

[0797] Ab-((Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D) n ;

[0798] Ab-((Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D) n ;

[0799] Ab-((Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D) n ;

[0800] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-D) n ;

[0801] Ab-((Succinimidyl-3-yl-N)-CH2-C(=O)-P1-P2-PAB-D)n ;

[0802] Ab-((Succinimidyl-3-yl-N)-CH2CH2-C(=O)-P1-P2-PAB-D) n ;

[0803] Ab-((Succinimidyl-3-yl-N)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D) n ;

[0804] Ab-((Succinimidyl-3-yl-N)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D) n ;

[0805] Ab-((Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D) n ;

[0806] Ab-((Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D) n ;

[0807] Ab-((Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D) n ;

[0808] Ab-((Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D) n ;

[0809] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-PAB-D) n ;

[0810] The Ab represents a monoclonal antibody or an antigenic structural fragment thereof, or a modified monoclonal antibody or an antigen-binding fragment thereof, preferably an anti-Her2, anti-TF, anti-ROR1, anti-GPC3 antibody or an antigenic structural fragment thereof;

[0811] The PAB has the following structure:

[0812] The left side Indicates the connection position with P2, on the right Indicates the location of connection to the drug unit;

[0813] The -P1-P2- represents the following structure:

[0814] -His-Val-Leu-Asn-Leu-Val-Cit-(SEQ ID NO: 1);

[0815] -Val-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:2);

[0816] -Ile-Pro-Val-Ser-Leu-Val-Cit-(SEQ ID NO:3);

[0817] -Gly-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:4);

[0818] -Gly-Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:5);

[0819] -Gly-Pro-Ala-Gly-Leu-Val-Cit-(SEQ ID NO: 6);

[0820] -Gly-Pro-Gln-Gly-Leu-Val-Cit-(SEQ ID NO:7);

[0821] -Gly-Pro-Leu-Gly-Leu-Val-Cit-(SEQ ID NO:8);

[0822] -Gly-Pro-Ser-Gly-Leu-Val-Cit-(SEQ ID NO:9);

[0823] -Gly-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO: 10);

[0824] -Gly-Pro-Ala-Ala-Leu-Val-Cit-(SEQ ID NO: 11);

[0825] -Gly-Pro-Ser-Ala-Leu-Val-Cit-(SEQ ID NO:12);

[0826] -His-Val-Leu-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:13);

[0827] -Val-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:14);

[0828] -Ile-Pro-Val-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:15);

[0829] -Gly-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:16);

[0830] -Gly-Pro-Ala-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:17);

[0831] -Gly-Pro-Ala-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:18);

[0832] -Gly-Pro-Gln-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:19);

[0833] -Gly-Pro-Leu-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:20);

[0834] -Gly-Pro-Ser-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:21);

[0835] -Gly-Pro-Arg-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:22);

[0836] -Gly-Pro-Ala-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:23);

[0837] -Gly-Pro-Ser-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:24);

[0838] -Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:25);

[0839] -Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:26);

[0840] -Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:27);

[0841] -Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:28);

[0842] -Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:29);

[0843] -Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:30);

[0844] -Gly-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:31);

[0845] -Gly-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:32);

[0846] -Gly-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:33);

[0847] -Gly-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:34);

[0848] -Gly-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:35);

[0849] -His-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:36);

[0850] -His-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:37);

[0851] -His-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:38);

[0852] -His-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:39);

[0853] -His-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:40);

[0854] -His-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:41);

[0855] -Ser-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:42);

[0856] -Ser-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:43);

[0857] -Ser-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:44);

[0858] -Ser-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:45);

[0859] -Ser-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:46);

[0860] -Ser-Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:47);

[0861] -Ser-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:48);

[0862] -Gly-Pro-Ala-Pro-Leu-Val-Cit-(SEQ ID NO:49);

[0863] -Gly-Pro-Ala-Asn-Leu-Val-Cit-(SEQ ID NO:50);

[0864] -Gly-Pro-Arg-Gly-Dap(Me2)-Val-Cit-(SEQ ID NO:51);

[0865] -Gly-Pro-Arg-Asn-Dap(Me2)-Val-Cit-(SEQ ID NO:60);

[0866] -Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:61);

[0867] -Gly-Pro-Arg-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO: 62);

[0868] -Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:63);

[0869] -Gly-Pro-Cit-Asn-Leu-Val-Cit-(SEQ ID NO: 64);

[0870] -Ser-Pro-Cit-Ser-Dap(Me2)-Val-Cit-(SEQ ID NO:65);

[0871] -Pro-Arg-Ser-Dap(Me2)-Val-Cit-(SEQ ID NO:66);

[0872] -Pro-Cit-Asn-Leu-Val-Cit-(SEQ ID NO:67);

[0873] -Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:68);

[0874] or a variant having one or two mismatches relative to the sequence of any one of SEQ ID NOs: 1-51 and 60-68;

[0875] The D is camptothecin or its derivatives, auristatin or its derivatives, tetrahydroisoquinoline alkaloids or its derivatives, protein degraders and their derivatives, preferably DX8951, MMAE, Dxd, SN38, ET743, Lurbinectedin, CC885, CC-90009;

[0876] Said n represents an integer of 1-10.

[0877] In some embodiments, the antibody drug conjugate is selected from:

[0878] In some embodiments, the antibody drug conjugate is selected from the following structures:

[0879] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-P1-P2-D) n ;

[0880] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-P1-P2-PAB-D) n ;

[0881] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-P1-P2-PAB-DMEDA-D) n ;

[0882] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-P1-P2-NMEDA-D) n ;

[0883] Ab-((C=O)-C6alkylene-C(=O)-D) n ;

[0884] Ab-((C=O)-C6alkylene-C(=O)-P1-P2-PAB-D) n ;

[0885] Ab-((C=O)-C6alkylene-C(=O)-P1-P2-PAB-DMEDA-D) n ;

[0886] Ab-((C=O)-C6alkylene-C(=O)-P1-P2-NMEDA-D) n ;

[0887] Ab-((Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-P1-P2-D) n ;

[0888] Ab-((Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-P1-P2-PAB-D) n ;

[0889] Ab-((Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-P1-P2-PAB-DMEDA-D) n ;

[0890] Ab-((Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-P1-P2-NMEDA-D) n ;

[0891] The Ab represents a monoclonal antibody or an antigenic fragment thereof, or a modified monoclonal antibody or an antigen-binding fragment thereof, preferably an anti-Her2, anti-TF, anti-ROR1 or anti-GPC3 antibody or an antigenic fragment thereof; the D represents camptothecin or a derivative thereof, auristatin or a derivative thereof, a tetrahydroisoquinoline alkaloid or a derivative thereof, a protein degrader and a derivative thereof, preferably DX8951, MMAE, Dxd, SN38, ET743, Lurbinectedin, CC885, CC-90009; the n represents any value selected from 1-10, preferably 2-9 or 3-8 (for example, 2, 3, 4, 5, 6, 7, 8, 9).

[0892] In some embodiments, the antibody drug conjugate is selected from the following structures:

[0893] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAGLVCit-PAB-D) n ;

[0894] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPLGLGGFG-D) n ;

[0895] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPASLVCit-PAB-D) n ;

[0896] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRGLVCit-PAB-D) n ;

[0897] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPSALVCit-PAB-D) n ;

[0898] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRGLGGFG-D) n ;

[0899] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAALGGFG-D) n ;

[0900] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRNLVCit-PAB-D) n ;

[0901] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLVCit-PAB-D) n ;

[0902] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNDap(Me2)VCit-PAB-D) n ;

[0903] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRNLVCit-PAB-D) n ;

[0904] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPLSLVCit-PAB-D) n ;

[0905] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRGLVCit-PAB-D) n ;

[0906] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRPLVCit-PAB-D) n ;

[0907] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRSLVCit-PAB-D) n ;

[0908] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRPLVCit-PAB-D) n ;

[0909] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPCitPLVCit-PAB-D) n ;

[0910] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-HPRGLVCit-PAB-D) n ;

[0911] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-HPCitPLVCit-PAB-D) n ;

[0912] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitGLVCit-PAB-D) n ;

[0913] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAPLVCit-PAB-D) n ;

[0914] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PLSLVCit-PAB-D) n ;

[0915] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitSLVCit-PAB-D) n ;

[0916] Ab-((C=O)-C6alkylene-C(=O)-GPRNLVCit-PAB-D) n ;

[0917] Ab-((Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-GPRNLVCit-PAB-D) n ;

[0918] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLVCit-PAB-DMEDA-D) n ;

[0919] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLGGFG-D) n ;

[0920] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLGGFG-NMEDA-D) n ;

[0921] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRSLVCit-PAB-D) n ;

[0922] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRSLVCit-PAB-D) n ;

[0923] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PCitSLVCit-PAB-D) n ;

[0924] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PCitPLVCit-PAB-D) n ;

[0925] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PCitGLVCit-PAB-D) n ;

[0926] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPCitGLVCit-PAB-D) n ;

[0927] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRGLVCit-PAB-D) n ;

[0928] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRPLVCit-PAB-D) n ;

[0929] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitPLVCit-PAB-D) n ;

[0930] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPANLVCit-PAB-D) n ;

[0931] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPCitNLVCit-PAB-D) n ;

[0932] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitSDap(Me2)VCit-PAB-D) n ;

[0933] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRSDap(Me2)VCit-PAB-D) n ;

[0934] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PCitNLVCit-PAB-D) n ;as well as

[0935] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PASLVCit-PAB-D) n ;

[0936] The Ab is an anti-Her2, anti-TF, anti-ROR1 or anti-GPC3 antibody or an antigenic structural fragment thereof; the D is DX8951, MMAE, Dxd, SN38 or ET743; and the n represents any value selected from 2-9, preferably 2-8 (e.g., 2, 3, 4, 5, 6, 7, 8, 9).

[0937] In some embodiments, the antibody drug conjugate is selected from the following structures:

[0938] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAGLVCit-PAB-DX8951) n ;

[0939] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAGLVCit-PAB-MMAE) n ;

[0940] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAGLVCit-PAB-ET743) n ;

[0941] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPLGLGGFG-DX8951) n ;

[0942] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPASLVCit-PAB-DX8951) n ;

[0943] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRGLVCit-PAB-DX8951) n ;

[0944] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRGLVCit-PAB-MMAE) n ;

[0945] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPSALVCit-PAB-DX8951) n ;

[0946] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRGLGGFG-DX8951) n ;

[0947] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAALGGFG-DX8951) n ;

[0948] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRNLVCit-PAB-DX8951) n ;

[0949] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRNLVCit-PAB-MMAE) n ;

[0950] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLVCit-PAB-DX8951) n ;

[0951] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLVCit-PAB-MMAE) n ;

[0952] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLVCit-PAB-ET743) n ;

[0953] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNDap(Me2)VCit-PAB-DX8951) n ;

[0954] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNDap(Me2)VCit-PAB-MMAE) n ;

[0955] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-SPRNLVCit-PAB-DX8951) n ;

[0956] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRNLVCit-PAB-MMAE) n ;

[0957] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPLSLVCit-PAB-DX8951) n ;

[0958] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPLSLVCit-PAB-MMAE) n ;

[0959] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRGLVCit-PAB-DX8951) n ;

[0960] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRPLVCit-PAB-DX8951) n ;

[0961] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRSLVCit-PAB-DX8951) n ;

[0962] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRPLVCit-PAB-DX8951) n ;

[0963] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPCitPLVCit-PAB-DX8951) n ;

[0964] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-HPRGLVCit-PAB-DX8951) n ;

[0965] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-HPCitPLVCit-PAB-DX8951) n ;

[0966] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-SPCitGLVCit-PAB-DX8951) n ;

[0967] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPAPLVCit-PAB-DX8951) n ;

[0968] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PLSLVCit-PAB-DX8951) n ;

[0969] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-SPCitSLVCit-PAB-DX8951) n ;

[0970] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-SPCitSLVCit-PAB-MMAE) n ;

[0971] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-SPCitSLVCit-PAB-ET743) n ;

[0972] Ab-((C=O)-C6alkylene-C(=O)-GPRNLVCit-PAB-DX8951) n ;

[0973] Ab-((Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-GPRNLVCit-PAB-DX8951) n ;

[0974] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPRNLVCit-PAB-DMEDA-SN38) n ;

[0975] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLGGFG-Dxd) n ;

[0976] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLGGFG-NMEDA-Dxd) n ;

[0977] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRSLVCit-PAB-DX8951) n ;

[0978] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRSLVCit-PAB-DX8951) n ;

[0979] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-PCitSLVCit-PAB-DX8951) n ;

[0980] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-PCitPLVCit-PAB-DX8951) n ;

[0981] Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(═O)-PCitGLVCit-PAB-DX8951) n ;

[0982] Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(═O)-GPCitGLVCit-PAB-DX8951) n ;

[0983] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRGLVCit-PAB-DX8951) n ;

[0984] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRPLVCit-PAB-DX8951) n ;

[0985] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-SPCitPLVCit-PAB-DX8951) n ;

[0986] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPANLVCit-PAB-DX8951) n ;

[0987] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPCitNLVCit-PAB-DX8951) n ;

[0988] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-SPCitSDap(Me2)VCit-PAB-DX8951) n ;

[0989] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitSDap(Me2)VCit-PAB-MMAE) n ;

[0990] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRSDap(Me2)VCit-PAB-DX8951) n ;

[0991] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRSDap(Me2)VCit-PAB-MMAE) n ;

[0992] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-PCitNLVCit-PAB-DX8951) n ;as well as

[0993] Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PASLVCit-PAB-DX8951) n ;

[0994] The Ab is an anti-Her2, anti-TF, anti-ROR1 or anti-GPC3 antibody or an antigenic structural fragment thereof; the n represents any value selected from 2-9, preferably 2-8 (e.g., 2, 3, 4, 5, 6, 7, 8, 9).

[0995] In some embodiments, the antibody drug conjugate is selected from the following structures:

[0996] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAGLVCit-PAB-DX8951) n ;

[0997] Anti-GPC3 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAGLVCit-PAB-DX8951) n ;

[0998] Anti-Her2 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAGLVCit-PAB-DX8951) n ;

[0999] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAGLVCit-PAB-MMAE) n ;

[1000] Anti-GPC3 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAGLVCit-PAB-MMAE) n ;

[1001] Anti-GPC3 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAGLVCit-PAB-ET743) n ;

[1002] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPLGLGGFG-DX8951) n ;

[1003] Anti-Her2 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPLGLGGFG-DX8951) n ;

[1004] Anti-Her2 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPASLVCit-PAB-DX8951) n ;

[1005] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPRGLVCit-PAB-DX8951) n ;

[1006] Anti-GPC3 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPRGLVCit-PAB-DX8951) n ;

[1007] Anti-Her2 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPRGLVCit-PAB-DX8951) n ;

[1008] Anti-ROR1 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPRGLVCit-PAB-DX8951) n ;

[1009] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPRGLVCit-PAB-MMAE) n ;

[1010] Anti-GPC3 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPRGLVCit-PAB-MMAE) n ;

[1011] Anti-Her2 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPSALVCit-PAB-DX8951) n ;

[1012] Anti-Her2 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRGLGGFG-DX8951) n ;

[1013] Anti-Her2 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAALGGFG-DX8951) n ;

[1014] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRNLVCit-PAB-DX8951) n ;

[1015] Anti-GPC3 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-PRNLVCit-PAB-DX8951) n ;

[1016] Anti-Her2 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-PRNLVCit-PAB-DX8951) n ;

[1017] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-PRNLVCit-PAB-MMAE) n ;

[1018] Anti-GPC3 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-PRNLVCit-PAB-MMAE) n ;

[1019] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLVCit-PAB-DX8951) n ;

[1020] Anti-GPC3 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLVCit-PAB-DX8951) n ;

[1021] Anti-Her2 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPRNLVCit-PAB-DX8951) n ;

[1022] Anti-ROR1 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPRNLVCit-PAB-DX8951) n ;

[1023] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLVCit-PAB-MMAE) n ;

[1024] Anti-GPC3 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPRNLVCit-PAB-MMAE) n ;

[1025] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPRNDap(Me2)VCit-PAB-DX8951) n ;

[1026] Anti-GPC3 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPRNDap(Me2)VCit-PAB-DX8951) n ;

[1027] Anti-Her2 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPRNDap(Me2)VCit-PAB-DX8951) n ;

[1028] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPRNDap(Me2)VCit-PAB-MMAE) n ;

[1029] Anti-GPC3 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPRNDap(Me2)VCit-PAB-MMAE) n ;

[1030] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRNLVCit-PAB-DX8951) n ;

[1031] Anti-GPC3 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-SPRNLVCit-PAB-DX8951) n ;

[1032] Anti-Her2 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-SPRNLVCit-PAB-DX8951) n ;

[1033] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRNLVCit-PAB-MMAE) n ;

[1034] Anti-GPC3 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPLSLVCit-PAB-DX8951) n ;

[1035] Anti-GPC3 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPLSLVCit-PAB-MMAE) n ;

[1036] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-PRGLVCit-PAB-DX8951) n ;

[1037] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRPLVCit-PAB-DX8951) n ;

[1038] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRSLVCit-PAB-DX8951) n ;

[1039] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRPLVCit-PAB-DX8951) n ;

[1040] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPCitPLVCit-PAB-DX8951) n ;

[1041] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-HPRGLVCit-PAB-DX8951) n ;

[1042] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PLSLVCit-PAB-DX8951) n ;

[1043] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-SPCitSLVCit-PAB-DX8951) n ;

[1044] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-SPCitSLVCit-PAB-MMAE) n ;

[1045] Anti-Her2 Antibody-((C=O)-C6 Alkylene-C(=O)-GPRNLVCit-PAB-DX8951) n ;

[1046] Anti-Her2 Antibody-((Succinimidyl-3-yl-N)-C2alkylene-C(═O)NH-(CH2CH2O)8-C2alkylene-C(═O)-GPRNLVCit-PAB-DX8951) n ;

[1047] Anti-Her2 Antibody-((Succinimidyl-3-yl-N)-C5 Alkylene-C(═O)-GPRNLVCit-PAB-DMEDA-SN38)n ;

[1048] Anti-Her2 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLGGFG-Dxd) n ;

[1049] Anti-Her2 Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPRNLGGFG-NMEDA-Dxd) n ;

[1050] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-PRSLVCit-PAB-DX8951) n ;

[1051] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRSLVCit-PAB-DX8951) n ;

[1052] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-PCitSLVCit-PAB-DX8951) n ;

[1053] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-PCitPLVCit-PAB-DX8951) n ;

[1054] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-PCitGLVCit-PAB-DX8951) n ;

[1055] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5 Alkylene-C(═O)-GPCitGLVCit-PAB-DX8951) n ;

[1056] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-SPRGLVCit-PAB-DX8951) n ;

[1057] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRPLVCit-PAB-DX8951) n ;

[1058] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-SPCitPLVCit-PAB-DX8951) n ;

[1059] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPANLVCit-PAB-DX8951) n ;

[1060] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-GPCitNLVCit-PAB-DX8951) n ;

[1061] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-SPCitSDap(Me2)VCit-PAB-DX8951) n ;

[1062] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-SPCitSDap(Me2)VCit-PAB-MMAE) n ;

[1063] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-PRSDap(Me2)VCit-PAB-DX8951) n ;

[1064] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-PRSDap(Me2)VCit-PAB-MMAE) n ;

[1065] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(═O)-PCitNLVCit-PAB-DX8951) n ;as well as

[1066] Anti-TF Antibody-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PASLVCit-PAB-DX8951) n ;

[1067] Said n represents any value selected from 2-9, preferably 2-8 (eg, 2, 3, 4, 5, 6, 7, 8, 9); said anti-Her2, anti-TF, anti-ROR1 or anti-GPC3 antibody is as described herein.

[1068] In some embodiments, the anti-Her2, anti-TF, anti-ROR1 and anti-GPC3 antibodies are selected from Trastuzumab, Tisotumab, Zilovertamab, Codrituzumab.

[1069] In some embodiments, the antibody drug conjugate is selected from the following structures:

[1070] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-DX8951)n;

[1071] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLGLGGFG-DX8951)n;

[1072] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPASLVCit-PAB-DX8951)n;

[1073] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLVCit-PAB-DX8951)n;

[1074] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPSALVCit-PAB-DX8951)n;

[1075] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLGGFG-DX8951)n;

[1076] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAALGGFG-DX8951)n;

[1077] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-PRNLVCit-PAB-DX8951)n;

[1078] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRNLVCit-PAB-DX8951)n;

[1079] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGDap(Me2)LVCit-PAB-DX8951)n;

[1080] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-SPRNLVCit-PAB-DX8951)n;

[1081] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLSLVCit-PAB-DX8951)n;

[1082] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-MMAE)n;

[1083] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLVCit-PAB-MMAE)n;

[1084] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-PRNLVCit-PAB-MMAE)n;

[1085] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRNLVCit-PAB-MMAE)n;

[1086] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGDap(Me2)LVCit-PAB-MMAE)n;

[1087] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-SPRNLVCit-PAB-MMAE)n;

[1088] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLSLVCit-PAB-MMAE)n;

[1089] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-ET743)n;

[1090] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRNLVCit-PAB-ET743)n;

[1091] The Ab represents a monoclonal antibody or an antigenic structural fragment thereof, or a modified monoclonal antibody or an antigen-binding fragment thereof, preferably an anti-Her2, anti-TF, anti-ROR1, anti-GPC3 antibody or an antigenic structural fragment thereof;

[1092] The DX8951 has the following structure:

[1093] in Indicates the connection position of DX8951 and the linker;

[1094] The MMAE has the following structure:

[1095] in Indicates the connection position between MMAE and the linker;

[1096] The ET-743 has the following structure:

[1097] in represents the connection position of ET743 and the linker; n is an integer from 1 to 10. Preferably, n is 2, 4, 6, or 8.

[1098] In another aspect, the present application provides a monoclonal antibody.

[1099] An anti-tissue factor (TF) antibody or an antigen-binding fragment thereof, comprising:

[1100] (a) VH comprising the amino acid sequence of SEQ ID NO: 52 or 54, or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 52 or 54, or an amino acid sequence obtained by substitution, deletion, or addition of one or more amino acids to the amino acid sequence of SEQ ID NO: 52 or 54, which functions the same or similarly to the amino acid sequence of SEQ ID NO: 52 or 54; and / or

[1101] (b) VL, which comprises the amino acid sequence of SEQ ID NO: 53 or 55, or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 53 or 55, or an amino acid sequence obtained by substitution, deletion or addition of one or more amino acids to the amino acid sequence of SEQ ID NO: 53 or 55, and which functions the same or similarly to the amino acid sequence of SEQ ID NO: 53 or 55.

[1102] In some embodiments, the anti-TF antibody or antigen-binding fragment thereof comprises:

[1103] (1) VH and VL, wherein the VH has the amino acid sequence shown in SEQ ID NO: 52, and the VL has the amino acid sequence shown in SEQ ID NO: 53; or

[1104] (2) VH and VL, wherein the VH has the amino acid sequence shown in SEQ ID NO: 54, and the VL has the amino acid sequence shown in SEQ ID NO: 53; or

[1105] (3) VH and VL, wherein the VH has the amino acid sequence shown in SEQ ID NO: 54, and the VL has the amino acid sequence shown in SEQ ID NO: 55; or

[1106] (4) VH and VL, wherein the VH has the amino acid sequence shown in SEQ ID NO: 52, and the VL has the amino acid sequence shown in SEQ ID NO: 55.

[1107] In some embodiments, the anti-TF antibody or antigen-binding fragment thereof comprises a light chain as set forth in SEQ ID NO: 56, or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 56; and a heavy chain as set forth in SEQ ID NO: 57, or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 57.

[1108] In some embodiments, the anti-TF antibody or antigen-binding fragment thereof comprises a light chain as set forth in SEQ ID NO: 56, or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 56; and a heavy chain as set forth in SEQ ID NO: 59, or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 59.

[1109] In some embodiments, the anti-TF antibody or antigen-binding fragment thereof comprises a light chain as set forth in SEQ ID NO: 58, or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 58; and a heavy chain as set forth in SEQ ID NO: 59, or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 59.

[1110] In some embodiments, the TF is mammalian TF, and the antibody is preferably a mouse antibody, a monkey antibody, a rabbit antibody, a chimeric antibody, a humanized antibody, or a human antibody; and / or

[1111] The antibody is an IgG antibody, preferably an IgG1, IgG2, IgG3 or IgG4 antibody, more preferably an IgG1 antibody; and / or

[1112] The anti-TF antibody or antigen-binding fragment thereof is selected from the group consisting of a monoclonal antibody, a polyclonal antibody, a Fab fragment, a Fab' fragment, a F(ab')2 fragment, a Fd fragment, a Fv fragment, a dAb fragment, an isolated CDR region, a scFv and a nanobody.

[1113] In some embodiments, the TF is mammalian TF, and the antibody is preferably a modified mouse antibody, monkey antibody, rabbit antibody, chimeric antibody, humanized antibody, or human antibody; and / or

[1114] The antibody is a modified IgG antibody, preferably a modified IgG1, IgG2, IgG3 or IgG4 antibody, more preferably a modified IgG1 antibody; and / or

[1115] The anti-TF antibody or antigen-binding fragment thereof is selected from the group consisting of modified monoclonal antibodies, polyclonal antibodies, Fab fragments, Fab' fragments, F(ab')2 fragments, Fd fragments, Fv fragments, dAb fragments, isolated CDR regions, scFv and nanobodies.

[1116] In some embodiments, the modification methods include: amino acid point mutation, glycosylation modification, and polypeptide modification.

[1117] A nucleic acid molecule encoding the anti-TF antibody or antigen-binding fragment thereof, or the heavy chain variable region and / or light chain variable region thereof according to any one of the embodiments.

[1118] A recombinant vector comprising the nucleic acid molecule described in some embodiments.

[1119] A recombinant cell comprising the nucleic acid molecule described in some embodiments or a recombinant vector comprising the nucleic acid molecule.

[1120] A method for producing an anti-TF antibody or an antigen-binding fragment thereof, comprising culturing the recombinant cell described in some embodiments.

[1121] Use of an anti-TF antibody or antigen-binding fragment thereof in some embodiments, a nucleic acid molecule in some embodiments, a recombinant vector in some embodiments, or a recombinant cell in some embodiments in the preparation of a medicament; preferably, the medicament is a medicament for preventing and / or treating cancer; more preferably, the cancer is a TF-expressing cancer.

[1122] Use of the anti-TF antibody or antigen-binding fragment thereof in some embodiments, the nucleic acid molecule in some embodiments, the recombinant vector in some embodiments, or the recombinant cell in some embodiments in the preparation of a reagent for diagnosing a cell proliferation-related disease; preferably, the disease is cancer; more preferably, the cancer is a TF-expressing cancer.

[1123] On the other hand, the present application provides preparation processes and uses of polypeptides or polypeptide fragments, linkers, linker drugs, and conjugates.

[1124] A method for preparing the polypeptide or polypeptide fragment -P1-P2- or its tautomers, mesomers, enantiomers, diastereomers, or mixed forms thereof, or an acceptable salt thereof, or a precursor thereof in some embodiments.

[1125] A method for preparing a linker as described in some embodiments, or a tautomer, mesomer, enantiomer, diastereomer, or mixture thereof, or an acceptable salt thereof, or a precursor thereof, comprising sequentially linking a polypeptide or polypeptide fragment -P1-P2- as described in some embodiments to its preceding and following units C1, La, L1, and L2. C1, La, L1, and L2 are as defined in some embodiments.

[1126] A method for preparing a linker drug as described in some embodiments, or a tautomer, mesomer, enantiomer, diastereomer, or mixture thereof, or an acceptable salt thereof, or a precursor thereof, comprises sequentially linking the polypeptide or polypeptide fragment -P1-P2- as described in some embodiments to its preceding and following units C1', La, L1, L2, and D. C1', La, L1, L2, and D are as defined in some embodiments.

[1127] A method for preparing the conjugate described in some embodiments or its tautomer, mesomer, enantiomer, diastereomer, or mixed form, or an acceptable salt thereof, or a precursor thereof, comprising sequentially connecting Bp, C1 or C1', La, L1, L2, P1, P2, D or a precursor thereof,

[1128] Wherein said P1 and P2 are defined in some embodiments;

[1129] C1, La, L1, L2 are as defined in some embodiments;

[1130] C1', D are defined in some embodiments;

[1131] Bp is defined in some embodiments.

[1132] A method for preparing the conjugate described in some embodiments or its tautomer, mesomer, enantiomer, diastereomer, or mixed form thereof, or an acceptable salt thereof, or a precursor thereof, comprises coupling Bp with a linker drug C1'-La-L1-P1-P2-L2-D in a suitable solvent and conditions, wherein C1'-La-L1-P1-P2-L2-D is defined in some embodiments.

[1133] Preferably, the molar ratio of Bp to the linker drug C1'-La-L1-P1-P2-L2-D is 1:(1-20); more preferably, the molar ratio of Bp to the linker drug C1'-La-L1-P1-P2-L2-D is 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, or 1:12.

[1134] Preferably, the coupling reaction is carried out in water or a buffer salt system and / or an organic solvent.

[1135] Preferably, the buffer salt system is selected from PBS, sodium acetate buffer, histidine-hydrochloric acid buffer, histidine-acetate buffer, citric acid buffer, and the above buffers added with sucrose; more preferably, the buffer is selected from PBS pH 7.4, PBS pH 7.2, PBS pH 6.5, PBS pH 6.0, histidine-acetate pH 6.5, histidine-acetate pH 6.0, histidine-acetate pH 5.5 or a combination with 5-9% sucrose.

[1136] Preferably, the organic solvent is selected from N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, N-methylpyrrolidone, acetonitrile, methanol, ethanol, ethylene glycol, propylene glycol, and glycerol.

[1137] Preferably, the method further comprises the step of purifying the coupled product. Preferably, the purification is performed using one or more of an ultrafiltration system, gel chromatography, affinity layer, hydrophobic chromatography, reverse phase chromatography, or ion exchange chromatography. More preferably, the purification is performed using one or more of an ultrafiltration tube, a tangential flow system, a desalting column, a gel column, a cation exchange column, an anion exchange column, or an affinity column.

[1138] A pharmaceutical composition comprising the polypeptide or polypeptide fragment of some embodiments, the linker of any one of some embodiments, the linker-drug of any one of some embodiments, the conjugate of any one of some embodiments, the antibody or antigen-binding fragment thereof of any one of some embodiments, the nucleic acid molecule of some embodiments, the recombinant vector of some embodiments, the recombinant cell of some embodiments, and tautomers, meso-racemates, racemates, enantiomers, diastereomers, or useful salts thereof, and optionally one or more pharmaceutical excipients. Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and / or excipient.

[1139] In some embodiments, the polypeptide or polypeptide fragment of any one of the embodiments, the linker of any one of the embodiments, the linker drug of any one of the embodiments, the conjugate of any one of the embodiments, the antibody or antigen-binding fragment thereof of any one of the embodiments, the nucleic acid molecule described in some embodiments, the recombinant vector described in some embodiments, the recombinant cell described in some embodiments, and its tautomer, meso-racemate, racemate, enantiomer, diastereomer or its available salt in the preparation of a medicament for treating / preventing a disease or condition. Alternatively, the present application discloses the polypeptide or polypeptide fragment of any one of the embodiments, the linker of any one of the embodiments, the linker drug of any one of the embodiments, the conjugate of any one of the embodiments, the antibody or antigen-binding fragment thereof of any one of the embodiments, the nucleic acid molecule described in some embodiments, the recombinant vector described in some embodiments, the recombinant cell described in some embodiments, and its tautomer, meso-racemate, racemate, enantiomer, diastereomer or its available salt in the treatment / prevention of a disease or condition. Alternatively, the present application discloses any one of the polypeptides or polypeptide fragments in some embodiments, any one of the linkers in some embodiments, any one of the linker drugs in some embodiments, any one of the conjugates in some embodiments, any one of the antibodies or antigen-binding fragments thereof in some embodiments, any one of the nucleic acid molecules in some embodiments, any one of the recombinant vectors in some embodiments, any one of the recombinant cells in some embodiments, and their tautomers, meso-racemates, racemates, enantiomers, diastereomers or their available salts for treating / preventing diseases or conditions.

[1140] In some embodiments, the polypeptide or polypeptide fragment of any one of the embodiments, the linker of any one of the embodiments, the linker drug of any one of the embodiments, the conjugate of any one of the embodiments, the antibody or antigen-binding fragment thereof of any one of the embodiments, the nucleic acid molecule described in some embodiments, the recombinant vector described in some embodiments, the recombinant cell described in some embodiments, and its tautomer, meso-racemate, racemate, enantiomer, diastereomer or its available salt in the preparation of a medicament for treating / preventing a disease associated with abnormal cell activity. Alternatively, the present application discloses the use of the polypeptide or polypeptide fragment of any one of the embodiments, the linker of any one of the embodiments, the linker drug of any one of the embodiments, the conjugate of any one of the embodiments, the antibody or antigen-binding fragment thereof of any one of the embodiments, the nucleic acid molecule described in some embodiments, the recombinant vector described in some embodiments, the recombinant cell described in some embodiments, and its tautomer, meso-racemate, racemate, enantiomer, diastereomer or its available salt in the treatment / prevention of a disease associated with abnormal cell activity. Alternatively, the present application discloses any one of the polypeptides or polypeptide fragments in some embodiments for treating / preventing diseases associated with abnormal cell activity, any one of the linkers in some embodiments, any one of the linker drugs in some embodiments, any one of the conjugates in some embodiments, any one of the antibodies or antigen-binding fragments thereof in some embodiments, the nucleic acid molecules described in some embodiments, the recombinant vectors described in some embodiments, the recombinant cells described in some embodiments, and their tautomers, meso-racemates, racemates, enantiomers, diastereomers or their available salts.

[1141] In some embodiments, the present application discloses the use of the pharmaceutical composition in preparing a medicament for treating / preventing a disease or condition. Alternatively, the present application discloses the use of the pharmaceutical composition described in some embodiments in treating / preventing a disease or condition. Alternatively, the present application discloses the pharmaceutical composition described in some embodiments for treating / preventing a disease or condition.

[1142] In some embodiments, the present application discloses the use of the pharmaceutical composition in the preparation of a medicament for treating / preventing a disease associated with abnormal cell activity. Alternatively, the present application discloses the use of the pharmaceutical composition described in some embodiments in the treatment / prevention of a disease associated with abnormal cell activity. Alternatively, the present application discloses the pharmaceutical composition described in some embodiments for treating / preventing a disease associated with abnormal cell activity.

[1143] In some embodiments, the disease or condition comprises a tumor.

[1144] In some embodiments, the disease or condition includes solid tumors and hematological tumors.

[1145] A method for treating a disease or condition (e.g., a tumor) in a subject in need thereof, comprising administering to the subject an effective amount of a conjugate according to any one of the embodiments. In this context, the effective amount can be determined by a person skilled in the art based on the subject's age, sex, weight, health status, family medical history, disease progression, etc. The conjugates described herein can be administered to the subject in conventional dosage forms via conventional routes of administration (e.g., intramuscular injection, etc.). In some embodiments, the tumor is selected from tumors associated with the expression of the following proteins: EGFR, HER2, HER3, c-Met, claudin18.2, TROP-2, folate receptor alpha (FRα), ROR1, ROR2, BCMA, PSMA, CD19, CD20, CD22, CD30, CD33, CD37, CD46, CD48, CD56, CD79b, CD123, CD138, CD166, CD276, CEACAM5, SDC1, CD74, CD70, MUC1, MUC16, GUCY2C, MSLN, SCL34A2, FOLH1, TPBC, PVRL4, STEAP1, SCL44A4, NACM1, EDNRB, GPNMB, FGFR, SEZ6, Nectin-4, PD-L1, Tissue Factor(TF), B7-H3, B7-H4, LY6E, LIV-1, CDH4, CDH6, ITGB6, GPC1, GPC3, ENPP3, KAAG1, FZD7, SSTR2, DLK1, TMEFF1, TM4SF1, SLAMF7, DLL3, FLT3, TNFRSF10B, EPCAM, AXL, IL2RA, 5T4, FAP, ICAM-1, IGF-1R;

[1146] Preferably, selected from tumors associated with Her2 expression;

[1147] Preferably, selected from tumors associated with Trop2 expression;

[1148] Preferably, it is selected from tumors associated with EGFR expression;

[1149] Preferably, it is selected from tumors related to FRα expression;

[1150] Preferably, it is selected from tumors associated with TF expression;

[1151] Preferably, it is selected from tumors associated with ROR1 expression;

[1152] Preferably, the tumor is selected from tumors associated with GPC3 expression.

[1153] In some embodiments, the tumor is selected from the group consisting of esophageal cancer, brain cancer, lung cancer, epithelial cell cancer, bladder cancer, gastric cancer, ovarian cancer, pancreatic cancer, head and neck cancer, urothelial cancer, colon cancer, colorectal cancer, cervical cancer, endometrial cancer, rectal cancer, kidney cancer, prostate cancer, melanoma, liver cancer, gallbladder cancer, bile duct cancer, thyroid cancer, non-Hodgkin's lymphoma, multiple myeloma, and B-cell lymphoma.

[1154] Preferably, the cancer is selected from esophageal cancer, lung cancer, breast cancer, gastric cancer, urothelial cancer, bile duct cancer, prostate cancer, colorectal cancer, cervical cancer, and ovarian cancer.

[1155] Definition of terms

[1156] Unless otherwise defined, all technical terms, symbols, and other scientific terms used herein are intended to have the meanings commonly understood by those skilled in the art. In some cases, terms with commonly understood meanings are defined herein for clarity and / or as ready reference, and the inclusion of such definitions herein should not be construed as representing a difference from what is generally understood in the art. The techniques and procedures described or referenced herein are generally well understood and commonly used by those skilled in the art using conventional methods.

[1157] Where appropriate, procedures involving the use of commercially available kits and reagents are generally performed according to the protocols and conditions defined by the manufacturers, unless otherwise indicated. As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly indicates otherwise. The terms "including," "such as," and the like are intended to express inclusion without limitation, unless otherwise expressly indicated. As used herein, the term "comprising" also specifically includes embodiments of "consisting of" and "consisting essentially of" the elements, unless otherwise expressly indicated.

[1158] In this application, the terms "antibody drug conjugate (antibody-drug conjugate)", "antibody drug conjugate (antibody-drug conjugate)" and "ADC (antibody drug conjugate)" are used interchangeably and generally refer to one or more therapeutic compounds (e.g., Dxd) linked to one or more antibodies or antigen-binding fragments and are defined by the following general formula: Bp-(LD)p, where Bp = antibody or antigen-binding fragment, L = linker, D = drug unit, and p = the number of drug moieties per antibody or antigen-binding fragment, which can be any number from 1 to 10. In some embodiments, the linker L may include a cleavable portion between the antibody or antigen-binding fragment and the drug unit. Exemplary cleavable linkers are described and exemplified herein.

[1159] In this application, the term "antibody" or its "antigen-binding fragment" is generally used in its broadest sense and includes certain types of immunoglobulin molecules that contain one or more antigen-binding domains that specifically bind to an antigen or epitope. Antibody specificity specifically encompasses monoclonal antibodies, polyclonal antibodies, dimers, multimers, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments such as Fab', Fab, F(ab'2, single domain antibodies (dAbs), Fv, scFv (single chain Fv), linear antibodies, diabodies, etc.; as long as they show the desired biological activity (Miller et al (2003) Jour. of Immunology 170: 4854-4861). Antibodies can be mouse, human, or human. Antibodies may be derived from other species, such as humanized, chimeric antibodies, or derived from other species. Without limitation, an "antibody" may typically comprise a protein comprising at least two heavy chains (HC) and two light chains (LC) interconnected by disulfide bonds, or an antigen-binding fragment thereof. Each heavy chain comprises a heavy chain variable region (VH) and a heavy chain constant region. In some naturally occurring IgG, IgD, and IgA antibodies, the heavy chain constant region comprises three domains: CH1, CH2, and CH3. In some naturally occurring antibodies, each light chain comprises a light chain variable region (VL) and a light chain constant region. The variable regions contain one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, called complementarity determining regions (CDRs), which alternate with more conserved regions called framework regions (FRs). Each VH and VL contains three CDRs and four framework regions (FRs), arranged in the following order from amino-terminus to carboxyl-terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable domains of natural heavy and light chains each contain four FR regions (HFR1, HFR2, HFR3, HFR4, LFR5, and CDR6). The constant regions of antibodies are FR1, LFR2, LFR3, and LFR4, most of which adopt a B-sheet configuration and are connected by three CDRs, forming a loop connection and, in some cases, forming part of a β-sheet structure. The CDRs in each chain are closely approached together by the FR region and, together with the CDRs from the other chain, form the antigen-binding site of the antibody. The constant region of the antibody can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system.

[1160] In this application, the term "CDR" generally refers to one of the six hypervariable regions within the variable domain of an antibody that primarily contributes to antigen binding. One of the most commonly used definitions of the six CDRs is provided by Kabat EA et al., (1991) Sequences of proteins of immunological interest. NIH Publication 91-3242), Chothia et al., "Canonical Structures For the Hypervariable Regions of Immunoglobulins," J. Mol. Biol. 196:901 (1987); and MacCallum et al., "Antibody-Antigen Interactions: Contact Analysis and Binding Site Topography," J. Mol. Biol. 262:732 (1996). As used in this application, the Kabat definition of CDRs can be applied to CDR1, CDR2, and CDR3 (CDR L1, CDR L2, CDR L3, or L1, L2, L3) of a light chain variable domain, and CDR1, CDR2, and CDR3 (CDR H1, CDR H2, CDR H3) of a heavy chain variable domain. H3 or H1, H2, H3).

[1161] In the present application, term " variable " generally refers to such fact, and some part of the sequence of the variable domain of antibody changes strongly, and it forms the combination and specificity of various specific antibodies to its specific antigen. However, variability is not evenly distributed in the whole variable region of antibody. It is concentrated in three sections in light chain and heavy chain variable region, and is referred to as complementary determining region (CDR) or hypervariable region (HVR). The more highly conserved part in variable domain is referred to as framework (FR). In the art, the CDR of antibody can be defined by several methods, such as the Kabat definition rule based on sequence variability (see, Kabat et al., Immunological Protein Sequence, 5th edition, National Institutes of Health, Bethesda, Maryland (1991)), the Chothia definition rule based on structural loop region position (see, Al-Lazikani et al., J Mol Biol 273:927-48,1997) and the KABAT definition rule based on the concept of IMGT ontology (IMGT-ONTOLOGY) and IMGT Scientific chart rule.

[1162] In the present application, the term "monoclonal antibody" generally refers to an antibody obtained from a group of substantially homogeneous antibodies, i.e., the individual antibodies in the cluster are identical, except for a small amount of natural mutations that may exist. Monoclonal antibodies are generally highly specific for a single antigenic site. Moreover, different from conventional polyclonal antibody preparations (generally having different antibodies for different determinants), each monoclonal antibody is for a single determinant on the antigen. Except for their specificity, the advantage of monoclonal antibodies is that they can be synthesized by hybridoma culture and are not polluted by other immunoglobulins. The modifier "monoclonal" represents the feature of the antibody obtained from a substantially homogeneous antibody population, and is not to be construed as needing to produce antibodies by any ad hoc method. For example, the monoclonal antibody used herein can be prepared in hybridoma cells, or can be prepared by recombinant DNA methods.

[1163] In this application, the term "chimeric antibody" generally refers to an antibody in which the variable region is derived from one species and the constant region is derived from another species. Typically, the variable region is derived from an antibody of an experimental animal such as a rodent ("parent antibody"), and the constant region is derived from a human antibody, so that the resulting chimeric antibody is less likely to induce an adverse immune response in a human individual than the parent (e.g., mouse-derived) antibody.

[1164] In this application, the term "humanized antibody" generally refers to an antibody in which some or all of the amino acids outside the CDR region of a non-human antibody (e.g., a mouse antibody) are replaced by corresponding amino acids derived from human immunoglobulins. In the CDR region, small additions, deletions, insertions, replacements, or modifications of amino acids may also be permitted, as long as they still retain the ability of the antibody to bind to a specific antigen. A humanized antibody may optionally comprise at least a portion of a human immunoglobulin constant region. A "humanized antibody" retains antigenic specificity similar to that of the original antibody. The "humanized" form of a non-human (e.g., mouse) antibody may minimally comprise a chimeric antibody derived from a sequence of a non-human immunoglobulin. In some cases, the CDR region residues in a human immunoglobulin (recipient antibody) may be replaced with CDR region residues of a non-human species (donor antibody) (such as mouse, rat, rabbit, or non-human primate) with desired properties, affinity, and / or ability. In some cases, the FR region residues of a human immunoglobulin may be replaced with corresponding non-human residues. In addition, humanized antibodies may contain amino acid modifications that are not present in the recipient antibody or in the donor antibody. These modifications may be made to further improve antibody performance, such as binding affinity.

[1165] In this application, the terms "fully human antibodies", "full human antibodies" or "completely human antibodies", also known as "fully human monoclonal antibodies", are antibodies whose variable and constant regions are all human, eliminating immunogenicity and toxic side effects. The development of monoclonal antibodies has gone through four stages, namely: murine monoclonal antibodies, chimeric monoclonal antibodies, humanized monoclonal antibodies and fully human monoclonal antibodies. The antibodies or ligands described in this application can be fully human monoclonal antibodies. The relevant technologies for the preparation of fully human antibodies can be: human hybridoma technology, EBV-transformed B lymphocyte technology, phage display technology (phage display), transgenic mouse antibody preparation technology (transgenic mouse) and single B cell antibody preparation technology, etc. The terms "full-length antibody", "complete antibody" and "whole antibody" are used interchangeably herein to refer to antibodies having a structure that is substantially similar to a naturally occurring antibody and having a heavy chain comprising an Fc region. For example, when used to refer to an IgG molecule, a "full-length antibody" is an antibody comprising two heavy chains and two light chains.

[1166] As used herein, the term "Fc region" refers to the C-terminal region of an immunoglobulin heavy chain, which, in naturally occurring antibodies, interacts with Fc receptors and certain proteins of the complement system. The structures of the Fc regions of various immunoglobulins and the glycosylation sites contained therein are known in the art. J Allergy Clin Immunol, 2010, 125:S41-52, incorporated by reference in its entirety. The Fc region may be a naturally occurring Fc region or an Fc region modified as described elsewhere in the art or in this disclosure.

[1167] In this application, the term "antigen binding domain or binding fragment" refers to an antibody portion that can specifically bind to an antigen or epitope. An example of an antigen binding domain is an antigen binding domain formed by the VH-VL dimer of an antibody. Another example of an antigen binding domain is an antigen binding domain that is formed by the diversification of certain loops of the tenth fibronectin type III domain of a connexin. Antigen binding domains can be found in various situations, including antibodies and chimeric antigen receptors (CARs), such as CARs derived from antibodies or antibody fragments (such as scFvs). In this application, the term "ligand" generally refers to a macromolecular compound that can recognize and bind to an antigen or receptor associated with a target cell. The role of a ligand can be to present a drug to a target cell population bound to a ligand, and these ligands include but are not limited to protein hormones, lectins, growth factors, antibodies or other molecules that can bind to cells, receptors and / or antigens. In this application, the ligand may be represented by Ab. The ligand antigen forms a bond with the linker via a heteroatom on the ligand and may be an antibody or an antigen-binding fragment thereof. The antibody may be selected from a chimeric antibody, a humanized antibody, a fully human antibody, or a murine antibody; the antibody may be a monoclonal antibody. For example, the antibody may target the following targets: HER2, HER3, B7H3, TROP2, Claudin 18.2, CD30, CD33, CD70, or EGFR. For example, the antibody can be an antibody targeting the following targets: 5T4, AGS-16, ANGPTL4, ApoE, CD19, CTGF, CXCR5, FGF2, MCPT8, MFI2, MS4A7, NCA, Sema5b, SLITRK6, STC2, TGF, 0772P, 5T4, ACTA2, ADGREI, AG-7, AIF1, AKRIC1, AKRIC2, ASLG659, Axl, B7H3, BAFF-R, BCMA, BMPRIB, BNIP3, CIQA, CIQB, CA6, CADMI, CCD79b, CCL5, CCR5, CCR7, CD11e, CD123, CD138, CD1 42,CD147,CD166,CD19,CD19,CD22,CD21,CD20,CD205,CD22,CD223,CD228,CD25,CD30,CD33,CD37,CD38,CD40,CD45,CD45(PTPRC),CD46,CD47,CD49D(ITGA 4), CD56, CD66e, CD70, CD71, CD72, CD74, CD79a, CD79b, CD80, CDCP1, CDH11, CDIIb, CEA, CEACAMS, c-Met, COL6A3, COL7AI, CRIPTO, CSFIR, CTSD, CTSS, CXCL11,CXCL10,DDIT4,DLL3,DLL4,DR5,E16,EFNA4,EGFR,EGFRvIII,EGLN,EGLN3,EMR2,ENPP3,EpCAM,EphA2,EphB2R,ETB R,FcRH2,FcRHI,FGFR2,FGFR3,FLT3,FOLR-a,GD2,GEDA,GPC-1,GPNMB,GPR20,GZMB,HER2,HER3,HLADOB,HMOX1,IFI 6,IFNG,IGF-1R,IGFBP3,IL1ORA1,IL-13R,IL-2,IL20Ra,IL-3,IL-4,IL-6,IRTA2,KISSIR,KRT33A,LIV-1,LOX,LR P-1,LRRC15,LUM,LY64,LY6E,Ly86,LYPD3,MDP,MMP10,MMP14,MMP16,MPF,MSG783,MSLN,MUC-1,NaPi2b,Napi3b,Ne Ctin-4, Nectin-4, NOG, P2X5, pCAD, P-Cadherin, PDGFRA, PDK1, PD-L1, PFKFB3, PGF, PGK1, PIK3AP1, PIK3CD, PLOD2, PSCA, PSCAhlg, PSMA, PSMA, PTK7, P-Cadherin, RNF43, NaPi2b, ROR1, ROR2, SERPINE1, SLC39A6, SLTRK6, STAT1, STEAP1 , STEAP2, TCF4, TENB2, TGFB1, TGFB2, TGFBR1, TNFRSF21, TNFSF9, Trop-2, TrpM4, Tyro7, UPKIB, VEGFA, WNT5A, epidermal growth factor, brevican, mesothelin, sodium phosphate cotransporter 2B, claudin 18.2, endothrin receptor, mucins (such as mucin 1 and mucin 16), guanylate cyclase C, integrin α4β7, integrin αvβ6, trophoblast glycoprotein, or tissue factor.

[1168] In this application, the term "specific binding" refers to the interaction between a binding molecule (e.g., an antibody) and its binding partner (e.g., an antigen), and generally refers to the presence of a specific structure (e.g., an antigenic determinant or epitope) on the binding partner. In other words, even when the binding partner is present in a mixture of other molecules or organisms, the antibody will preferentially bind to or recognize the binding partner. Binding can be mediated by covalent or non-covalent interactions or a combination of the two. In other words, the term "specific binding" generally refers to immunospecific binding to an antigenic determinant or epitope and not immunospecific binding to other antigenic determinants or epitopes. The binding molecule of an immunospecific binding antigen can bind to other peptides or polypeptides with lower affinity, as measured by radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), BIACORE, or other analyses known in the art. The binding molecule of an immunospecific binding antigen or its fragment can cross-react with related antigens with the same epitope. In some cases, the binding molecule of an immunospecific binding antigen or its fragment does not cross-react with other antigens.

[1169] In this application, the term "affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen or epitope). Unless otherwise indicated, as used herein, "affinity" refers to intrinsic binding affinity, which reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen or epitope). The affinity of a molecule X for its partner Y can be expressed in terms of the dissociation equilibrium constant (K D ). The kinetic components that affect the dissociation equilibrium constant are described in more detail below. Affinity can be measured by common methods known in the art, including those described herein, such as surface plasmon resonance (SPR) technology (e.g. ) or biolayer interferometry (e.g. FORTEBIO).

[1170] In this application, the term "K d ”(sec -1 ) refers to the dissociation rate constant for a specific antibody-antigen interaction. This value is also known as K off value.

[1171] In this application, the term "k a ”(M -1 -×sec -1 ) refers to the association rate constant for a specific antibody-antigen interaction. This value is also called k on value.

[1172] In this application, the term "K D "(M), as used herein, refers to the dissociation equilibrium constant for a specific antibody-antigen interaction. KD =k d / k a In some embodiments, the K of the interaction between the antibody and its antigen is D Describes the affinity of an antibody. For clarity, as known in the art, a smaller K D A value indicates a higher affinity interaction, while a larger K D A value of 0 indicates a lower affinity interaction. If the K D Less than or equal to 5×10 -8 M, then the antibody is said to "specifically bind" to antigen X. More ideally, 1×10 -8 M or less, more ideally 6×10 -9 M or less, more preferably 3×10 -9 M or less, more preferably 2×10 -9 M or less. The antibody may be chimeric, humanized, or preferably human.

[1173] In this application, the term "K A "(M) refers to the binding equilibrium constant of a specific antibody-antigen interaction. K A =k a / k d .

[1174] In this application, the terms "linker", "linker", "L" or "linker structure" or "linker moiety" generally refer to a chemical structure fragment that is used to connect to a ligand at one end and to a cytotoxic drug at the other end, and can also be connected to other linkers before being used to connect to a cytotoxic drug. The direct or indirect connection to the ligand can mean that the group is directly connected to the ligand through a covalent bond, or it can be connected to the ligand through a linker structure. The linker may be susceptible to or substantially resist acid-induced cleavage, peptidase-induced cleavage, light-based cleavage, esterase-induced cleavage and / or disulfide bond cleavage under conditions that maintain the activity of the compound or antibody.

[1175] In this application, the term "polypeptide or fragment thereof" refers to a peptide structural unit comprising at least two covalently attached amino acids. The term encompasses polypeptides, oligopeptides and peptides. In some embodiments, two or more covalently attached amino acids are attached via peptide bonds and may be composed of naturally occurring amino acids and peptide bonds. Alternatively, synthetic amino acids (e.g., homophenylalanine, citrulline, ornithine and norleucine) or peptide mimetic structures, i.e., "peptide or protein analogs," such as peptoids, may be included. Peptoids are an exemplary class of peptide mimetics whose side chains are attached to nitrogen atoms of the peptide backbone, rather than α-carbons (as in amino acids), and have different hydrogen bonding and conformational characteristics compared to peptides. Thus, peptoids can resist proteolysis or other physiological or storage conditions and effectively penetrate cell membranes. In particular, when antibodies are synthesized in vitro by conventional methods well known in the art, such synthetic amino acids may be incorporated. In addition, any combination of peptide mimetic, synthetic and naturally occurring residues / structures may be used. "Amino acids" also include imino acid residues, such as proline and hydroxyproline. The amino acid "R group" or "side chain" can be in the (L)- or (S)-configuration. In a specific embodiment, the amino acid is in the (L)- or (S)-configuration.

[1176] In this application, the term "Drug unit" or "D" generally refers to any compound having the desired biological activity and reactive functional groups that can be used to incorporate the drug into the conjugate of the present application. In some embodiments, the Drug unit represents a cytotoxic drug for cancer treatment; a protein or polypeptide having the desired biological activity, such as abrin, ricin A, Pseudomonas exotoxin, and diphtheria toxin; other suitable proteins include tumor necrosis factor, α-interferon, β-interferon, nerve growth factor, platelet-derived growth factor, tissue plasminogen activator, and biological response modifiers, such as lymphokines, interleukin-1 (IL-1), interleukin-2 (IL-2), interleukin-6 (IL-6), granulocyte macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF), or other growth factors.

[1177] In this application, the term "cytotoxic drug" or "cytotoxic agent" generally refers to a toxic drug, which may refer to a substance that inhibits or prevents cell function and / or causes cell death or destruction. Cytotoxic drugs can kill tumor cells at sufficiently high concentrations. The "cytotoxic drug" may include cytotoxic drugs, including but not limited to radioactive isotopes (e.g., At 211 , I 131 , I 125 、Y 90 、Re 186 、Re 188 、Sm 153 、Bi212 、P 32 , Pb 212 and radioactive isotopes of Lu); chemotherapeutic agents or drugs (e.g., methotrexate, doxorubicin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunomycin or other intercalating agents); growth inhibitors; enzymes and fragments thereof, such as nucleotide degrading enzymes; antibiotics; toxins, such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and / or variants thereof; and various anti-tumor or anti-cancer agents disclosed below.

[1178] In this application, the term "isotopically labeled compound" or "isotopically labeled derivative" generally refers to a presently disclosed compound in which one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature, including pharmaceutical salts and prodrugs of each as described herein. Examples of isotopes that can be introduced into the presently disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as 2 H. 3 H. 13 C. 14 C. 15 N. 18 O. 17 O. 31 P. 32 P. 35 S. 18 F and 36 Cl.

[1179] In this application, the term "pharmaceutically acceptable form" generally refers to the form of the disclosed compound, including but not limited to pharmaceutically acceptable salts, esters, hydrates, solvates, polymorphs, isomers, prodrugs and isotopically labeled derivatives thereof. In one embodiment, "pharmaceutically acceptable form" includes but is not limited to pharmaceutically acceptable salts, esters, prodrugs and isotopically labeled derivatives thereof. In some embodiments, "pharmaceutically acceptable form" includes but is not limited to pharmaceutically acceptable isomers and stereoisomers, prodrugs and isotopically labeled derivatives thereof.

[1180] The linkers, linker drugs, and conjugates of the present application may include any pharmaceutically acceptable form thereof, such as pharmaceutically acceptable salts, esters, hydrates, solvates, polymorphs, isomers, prodrugs, or isotope-labeled derivatives. The above pharmaceutically acceptable forms are also within the scope of the present application.

[1181] In the present application, the term "pharmaceutically acceptable salt" generally refers to a pharmaceutically acceptable organic or inorganic salt of a compound (e.g., a drug, a linker drug, or a conjugate). In some aspects, the compound may contain at least one amino group and therefore may form an acid addition salt with the amino group. Exemplary salts include, but are not limited to, sulfate, trifluoroacetate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate, sucrose salt, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylenebis-(2-hydroxy-3-naphthoate)). A pharmaceutically acceptable salt may involve the introduction of another molecule such as an acetate ion, a succinate ion, or other counterion. The counterion can be any organic or inorganic moiety that will stabilize the charge on the parent compound. In addition, a pharmaceutically acceptable salt may have more than one charged atom in its structure. In cases where multiple charged atoms are part of a pharmaceutically acceptable salt, there may be multiple counterions. Thus, a pharmaceutically acceptable salt may have one or more charged atoms and / or one or more counterions.

[1182] In this application, the term "derivative" generally refers to a chemical compound or molecule made from a parent compound by one or more chemical reactions.

[1183] As used herein, the term "prodrug" generally refers to a less biologically active or inactive compound that is converted in vivo to a more biologically active compound via a chemical or biological process (ie, a chemical reaction or enzymatic biotransformation).

[1184] In this application, the term "pharmaceutically acceptable carrier" generally refers to one or more non-toxic substances that do not interfere with the effectiveness of the biological activity of the active ingredient. Such formulations may generally contain salts, buffers, preservatives, compatible carriers, adjuvants, and optionally other therapeutic agents. Such pharmaceutically acceptable formulations may also generally include compatible solid or liquid fillers, diluents, or encapsulating materials suitable for administration to humans. Pharmaceutically acceptable carriers may include, without limitation, liquids such as water, saline, glycerol, and ethanol. Auxiliary substances, such as wetting agents or emulsifiers, pH buffering substances, etc., may also be present in these carriers.

[1185] In this application, the term "methylene" generally refers to a residue derived from a group of one carbon atom by removing two hydrogen atoms. A methylene group may be substituted or unsubstituted, substituted or unsubstituted.

[1186] The term "alkylene" generally refers to a saturated straight or branched aliphatic hydrocarbon group having two residues derived from the same carbon atom or two different carbon atoms of a parent alkyl group by removing two hydrogen atoms, and can be a straight or branched group containing 1 to 20 carbon atoms, for example, an alkylene group containing 1 to 12 carbon atoms, for example, an alkylene group containing 1 to 6 carbon atoms. Non-limiting examples of alkylene include, but are not limited to, methylene (-CH2-), 1,1-ethylene (-CH(CH3)-), 1,2-ethylene (-CH2CH2)-, 1,1-propylene (-CH(CH2CH3)-), 1,2-propylene (CH2CH(CH3)-), 1,3-propylene (-CH2CH2CH2-), 1,4-butylene (-CH2CH2CH2CH2-), and 1,5-butylene (-CH2CH2CH2CH2CH2-). Alkylene may be substituted or unsubstituted, substituted or unsubstituted, for example, when substituted, the substituent may be substituted at any available point of attachment, preferably independently selected from one or more substituents selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio and oxo, for example, hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, or a C1-6 aliphatic group.

[1187] In this application, the term "arylene" generally refers to a radical having two hydrogen atoms derived from the same carbon atom or two different carbon atoms of an aromatic ring.

[1188] The term "aromatic ring" may refer to a 6- to 14-membered, all-carbon monocyclic ring or a fused polycyclic ring (i.e., rings that share adjacent pairs of carbon atoms) with a conjugated π-electron system. It may be 6- to 10-membered, such as benzene and naphthalene. The aromatic ring may be fused to a heteroaryl, heterocyclyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the aryl ring. The aryl group may be substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, thiol, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio.

[1189] The term "heteroaryl ring" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms can be selected from the group consisting of oxygen, sulfur, and nitrogen. Heteroaryl groups can be 5 to 10-membered, and can be 5- or 6-membered, such as furyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, and the like. The heteroaryl ring can be fused to an aryl, heterocyclyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the heteroaryl ring. Heteroarylene groups can be optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio.

[1190] The term "heteroaryl" refers to a group containing 5 or more ring atoms and exhibiting aromaticity, including heteroatoms. For example, "C2-C10 heteroaryl" refers to a group containing 2 to 10 carbon atoms in the ring atoms and the remainder being heteroatoms.

[1191] In this application, the term "heterocyclylene" generally refers to a stable 3-7 membered monocyclic ring structure without aromaticity, a fused 7-10 membered bicyclic heterocyclic ring structure, or a bridged 6-10 membered bicyclic heterocyclic ring structure. These ring structures may be saturated or partially saturated. In addition to carbon atoms, these ring structures may contain one or more heteroatoms, wherein the heteroatoms may be selected from the following group: oxygen, sulfur, and nitrogen. For example, it may contain 1-4 heteroatoms as defined above. When used to refer to atoms in a heterocyclic ring structure, the term "nitrogen" may include nitrogen that has undergone substitution reactions. Heterocyclylene may be substituted or unsubstituted.

[1192] In the present application, the term "heteroatom" refers to an atom other than C and H or a group containing the atom, such as N, O, S, P, NR, and the like.

[1193] In this application, the term "carbocyclylene" generally refers to a residue derived from the same carbon atom or two different carbon atoms of a carbocyclic ring by removing two hydrogen atoms. The term "carbocycle" generally refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon, the carbocyclic ring containing 3 to 20 carbon atoms, can contain 3 to 12 carbon atoms, can contain 3 to 10 carbon atoms, can contain 3 to 8 carbon atoms. Non-limiting examples of monocyclic carbocycles include cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexene, cyclohexadiene, cycloheptane, cycloheptatriene, cyclooctane, etc.; polycyclic carbocycles can include spirocyclic, fused ring and bridged ring carbocycles. Carbocyclylene can be substituted or unsubstituted.

[1194] As used herein, the term "partially unsaturated" generally refers to a cyclic structure containing at least one double or triple bond between the ring molecules. The term "partially unsaturated" encompasses cyclic structures with multiple unsaturations but is not intended to include aromatic or heteroaromatic rings as defined herein. The term "unsaturated" refers to a moiety having one or more degrees of unsaturation.

[1195] In the present application, the term "halogen" refers to any halogen group, typically fluorine (F), chlorine (Cl), bromine (Br), iodine (I), for example, fluorine and chlorine.

[1196] As used herein, the term "haloalkyl" refers to an alkyl group substituted with one or more halogen atoms.

[1197] In this application, the term "aliphatic group" generally refers to a straight-chain, branched-chain, or cyclic hydrocarbon having 1 to 12 carbon atoms, which is either fully saturated or has one or more unsaturated units, provided the unsaturated units are not aromatic. For example, suitable aliphatic groups may include substituted or unsubstituted straight-chain, branched, or cyclic alkyl, alkenyl, alkynyl groups, and mixtures thereof; such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, or (cycloalkyl)alkenyl groups. For example, aliphatic groups have 1 to 12, 1 to 8, 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[1198] In this application, the term "alkyl" or "alkyl group" generally refers to a fully saturated straight, branched, or cyclic hydrocarbon chain. In certain embodiments, the alkyl group may contain 1-8 carbon atoms ("C1-C8 alkyl"). In certain embodiments, the alkyl group may contain 1-6 carbon atoms ("C1-C6 alkyl"). In certain embodiments, the alkyl group contains 1-3 carbon atoms. In yet other embodiments, the alkyl group contains 2-3 carbon atoms, and in yet other embodiments, the alkyl group contains 1-2 carbon atoms.

[1199] In the present application, the term "alkylalkoxy" means an alkyl group substituted with an alkoxy group.

[1200] In this application, the term "alkoxy" refers to an alkyl group as previously defined attached to the main carbon chain via an oxygen ("alkoxy") atom.

[1201] In the present application, the term "alkylhydroxy" means an alkyl group substituted with a hydroxy group.

[1202] As used herein, the term "hydroxyl" refers to -OH.

[1203] In this application, the term "carbocycle" or "carbocyclyl" includes both aromatic groups (e.g., aryl) and non-aromatic groups (e.g., cycloalkyl). In certain embodiments, the carbocyclic group contains 3-10 carbon atoms ("3 to 10-membered carbocycle"). In certain embodiments, the carbocyclic group contains 3-8 carbon atoms ("3 to 8-membered carbocycle"). In certain embodiments, the carbocyclic group contains 3-6 carbon atoms ("3 to 6-membered carbocycle"). In certain embodiments, the carbocyclic group contains 3-5 carbon atoms ("3 to 5-membered carbocycle").

[1204] In this application, the terms "heterocycle" or "heterocyclyl" or "heterocyclic" mean a monocyclic, bicyclic, or tricyclic heterocycle containing at least one heteroatom in the ring. The terms "heterocycle" or "heterocyclyl" or "heterocyclic" encompass heteroaryl groups. "Heteroaryl" refers to a cyclic moiety having one or more closed rings and one or more heteroatoms (oxygen, nitrogen, or sulfur) in at least one of such rings, wherein at least one of such rings is aromatic, and wherein the ring or rings may independently be fused and / or bridged. Examples include, but are not limited to, phenyl, thienyl, triazolyl, pyridyl, pyrimidinyl, pyridazinyl, and pyrazinyl.

[1205] In this application, the term "PEG unit" generally refers to an organic moiety composed of repeating ethylene-oxy subunits (PEG or PEG subunits) and can be polydisperse, monodisperse or discrete (i.e., having a discrete number of ethylene-oxy subunits). Polydisperse PEG is a heterogeneous mixture of size and molecular weight, while monodisperse PEG is typically purified from a heterogeneous mixture and therefore has a single chain length and molecular weight. The PEG unit of the present application can comprise one or more polyethylene glycol chains, each composed of one or more ethyleneoxy subunits covalently attached to each other. The polyethylene glycol chains can be linked together, for example, in a linear, branched or star-shaped configuration.

[1206] As used herein, the term "bond" refers to a physical or chemical interaction between two substances or a combination thereof. Bonds include ionic bonds, nonionic bonds, hydrogen bonds, van der Waals bonds, hydrophobic interactions, and the like. Physical interactions (bonds) can be direct or indirect. An indirect physical interaction (bond) is mediated or caused by the action of another protein or compound. A direct bond refers to an interaction that does not occur through or as a result of the action of another protein or compound and does not substantially involve another intermediate.

[1207] In this application, the term "optional" or "optionally" generally means that the subsequently described event or circumstance can but need not occur, and the description includes instances where the event or circumstance occurs or does not occur. For example, "a heterocyclic group optionally substituted with an alkyl group" means that the alkyl group can but need not be present, and the description includes instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group.

[1208] In this application, the term "substituted" generally refers to one or more hydrogen atoms in a group, for example up to 5, for example 1 to 3 hydrogen atoms, which are independently replaced by a corresponding number of substituents. The substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) whether substitution is possible or not without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.

[1209] In the present application, the term "0 or more (e.g., 0 or at least 1, 0 or 1, 0) methylene units are "replaced" generally means that when the structure contains 1 or more methylene units, the one or more methylene units may be unsubstituted or replaced by one or more groups other than methylene (e.g., -NHC(O)-, -C(O)NH-, -C(O)-, -OC(O)-, -C(O)O-, -NH-, -O-, -S-, -SO-, -SO2-, -PH-, -P(-O)H-, -NHSO2-, -SONH-, -C(-S)-, -C(=NH)-, -N=N-, -C=N-, -N=C- or -C(=N2)-). One or more hydrogen atoms, for example up to 5, for example 1 to 3, in the group are independently substituted by a corresponding number of substituents. Substituents are only in their possible chemical positions, and one skilled in the art can determine (either experimentally or theoretically) which substitutions are possible or impossible without undue effort. For example, an amino or hydroxyl group having a free hydrogen may be unstable when combined with a carbon atom having an unsaturated (e.g., olefinic) bond.

[1210] In this application, the term "compound" generally refers to a substance having two or more different elements. For example, the compound of the present application can be an organic compound, for example, a compound of the present application can be a compound with a molecular weight of 500 or less, a compound with a molecular weight of 1000 or less, a compound with a molecular weight of 1000 or more, or a compound with a molecular weight of 10,000 or more, or a compound with a molecular weight of 10,000 or more, or a compound with a molecular weight of 100,000 or more. In this application, a compound can also refer to a compound connected by chemical bonds, for example, a compound in which one or more molecules with a molecular weight of 1000 or less are connected to a biomacromolecule by chemical bonds, wherein the biomacromolecule can be a polysaccharide, protein, nucleic acid, polypeptide, etc. For example, the compound of the present application can include a compound in which a protein is connected to one or more molecules with a molecular weight of 1000 or less, a compound in which a protein is connected to one or more molecules with a molecular weight of 100,000 or less, or a compound in which a protein is connected to one or more molecules with a molecular weight of 100,000 or less. Unless otherwise specified, the structures described in this application can also include compounds that differ only in the presence or absence of one or more isotopically enriched atoms. For example, compounds having the same structure as the present invention except that hydrogen atoms are replaced by deuterium or tritium, or carbon atoms are replaced by carbon 13 or carbon 14, are all within the scope of the present invention.

[1211] In the present application, the term "solvent", "solvent compound" or "buffer" generally refers to a pharmaceutically acceptable solvate formed by the ligand-drug conjugate compound of the present application and one or more solvent molecules. Non-limiting examples of solvent molecules include water, ethanol, acetonitrile, isopropanol, DMSO, ethyl acetate, etc.

[1212] In this application, the term "drug loading" generally refers to the average number of cytotoxic drugs loaded per ligand, and can also be expressed as the ratio of the cytotoxic drug to the antibody. The cytotoxic drug loading can range from 0 to 12 per ligand (Ab). In embodiments of this application, the drug loading is expressed as n", which can be an average of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. The drug loading of each ADC molecule after the conjugation reaction can be determined using conventional methods such as UV / visible spectroscopy, mass spectrometry, ELISA, and HPLC characterization.

[1213] One of ordinary skill in the art will understand that "substitution" or "substituted with" or "absence" includes the implicit limitation that the substitution or absence is according to the permitted valences of the substituted atom and substituents, and that the substitution or absence results in a stable compound, for example, the stable compound does not spontaneously undergo a transformation such as by rearrangement, cyclization, elimination, etc. For purposes of this disclosure, heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valences of the heteroatoms.

[1214] In this application, the term "prevention and / or treatment" includes not only preventing and / or treating a disease, but also generally includes preventing the onset of a disease, slowing or reversing the progression of a disease, preventing or slowing the onset of one or more symptoms associated with the disease, reducing and / or alleviating one or more symptoms associated with the disease, reducing the severity and / or duration of the disease and / or any symptoms associated therewith and / or preventing further increase in the severity of the disease and / or any symptoms associated therewith, preventing, reducing or reversing any physiological damage caused by the disease, and generally any pharmacological effect that is beneficial to the patient being treated. The compositions of the present application do not need to achieve a complete cure or eradication of any symptoms or manifestations of the disease in order to form a viable therapeutic agent. As recognized in the relevant art, drugs used as therapeutic agents can reduce the severity of a given disease state, but do not need to eliminate every manifestation of the disease to be considered a useful therapeutic agent. Similarly, a prophylactic treatment does not need to be completely effective in preventing the onset of the condition to constitute a viable preventive agent. It is sufficient to simply reduce the impact of the disease in the subject (e.g., by reducing the number or severity of its symptoms, or by increasing the effectiveness of another treatment, or by producing another beneficial effect), or to reduce the likelihood of the disease occurring or worsening.

[1215] In this application, the term "administering" generally refers to delivering a protein (including an immunoglobulin) to a human or animal in need thereof by any route known in the art. Pharmaceutical carriers and formulations or compositions are also well known in the art. Routes of administration may include intravenous, intramuscular, intradermal, subcutaneous, transdermal, mucosal, intratumoral, or mucosal.

[1216] In this application, the term "contacting" generally refers to the contacting of two or more different types of substances in any order, in any manner, and for any duration. When applied to cells, the term "contacting" generally refers to a method by which the antibodies or antigen-binding fragments thereof, drugs, linker drugs, conjugates, and / or pharmaceutical compositions of the present application are delivered to target cells or placed in direct proximity to target cells, which delivery can be in vitro or in vivo and can involve the use of recombinant vector systems. "Contacting" can occur in vivo, ex vivo, or in vitro.

[1217] In this application, the term "effective amount" or "effective dose" generally refers to an amount sufficient to achieve or at least partially achieve the desired effect. A "therapeutically effective amount" or "therapeutically effective dose" of a drug or therapeutic agent is generally any amount of the drug that promotes disease regression (as evidenced by a reduction in the severity of disease symptoms, an increase in the frequency and duration of disease symptom-free periods, or the prevention of damage or disability caused by the disease) when used alone or in combination with another therapeutic agent. A "prophylactic effective amount" or "prophylactic effective dose" of a drug generally refers to an amount of the drug that inhibits the development or recurrence of the disease when administered to a subject at risk of disease development or disease recurrence, either alone or in combination with another therapeutic agent. The ability of therapeutic or prophylactic agents to promote disease regression or inhibit disease development or recurrence can be assessed using a variety of methods known to those skilled in the art, such as in human subjects during clinical trials, predicting efficacy in humans in animal model systems, or by measuring the activity of the agent in in vitro assays.

[1218] In this application, the term "tumor" generally refers to all neoplastic cell growth and proliferation, whether malignant or benign, and all precancerous and cancerous cells and tissues. The terms "cancer," "cancer," "cell proliferative disorder," "proliferative disorder," and "tumor" are not mutually exclusive when referred to herein. In this application, a tumor can be a solid tumor or a non-solid tumor.

[1219] In this application, the term "subject" generally refers to a human or non-human animal (including mammals) in need of diagnosis, prognosis, improvement, prevention and / or treatment of a disease, such as humans, non-human primates (apes, gibbons, gorillas, chimpanzees, orangutans, macaques), livestock (dogs and cats), farm animals (poultry such as chickens and ducks, horses, cattle, goats, sheep, pigs) and experimental animals (mice, rats, rabbits, guinea pigs). Human subjects include fetuses, newborns, infants, adolescents and adult subjects. Subjects can include animal disease models.

[1220] In this application, the terms "comprises," "comprising," "having," "may," "containing," and variations thereof are generally intended to be open transitional phrases, terms, or words that do not exclude the possibility of additional actions or structures. The term "consisting of" generally indicates that no other components (or similarly, features, integers, steps, etc.) can be present. Unless the context clearly dictates otherwise, singular forms such as "a," "an," "the" in English, "one," "a kind," and "described / the" in Chinese generally include plural forms of the referents. Similarly, unless the context clearly indicates otherwise, plural terms herein encompass singular referents. Unless the context clearly indicates otherwise, the word "or" herein is intended to include "and."

[1221] In this application, the terms "host cell", "host cell line" and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced and the progeny of such cells. Host cells include "transformants" (or "transformed cells") and "transfectants" (or "transfected cells"), which both include primary transformed or transfected cells and progeny derived therefrom. Such progeny may not be completely identical to the parent cell in nucleic acid content and may contain mutations. Recombinant host cells include transfectomas, such as CHO cells, HEK293 cells, NS / 0 cells and lymphocytes. The term "transfectoma", as used herein, includes recombinant eukaryotic host cells, such as CHO cells, NS / 0 cells, HEK293 cells, plant cells or fungi (including yeast cells) that express antibodies.

[1222] As used herein, the term "vector" refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes vectors that are self-replicating nucleic acid structures, as well as vectors that are introduced into the genome of a host cell. Certain vectors are capable of directing the expression of nucleic acids to which they are operatively linked.

[1223] As used herein, the terms "inhibit" or "inhibition" mean a measurable reduction and can include, but do not require, complete prevention or inhibition. As used herein, "inhibit growth" or "inhibit proliferation" (e.g., of cells such as tumor cells) are intended to include any measurable reduction in cell growth when contacted with an anti-TF antibody drug conjugate, compared to the growth of the same cells not contacted with the anti-target cell antibody drug conjugate, for example, inhibition of growth of a cell culture by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 99%, or 100%. This reduction in cell growth can occur through a variety of mechanisms, which are acted upon by the anti-target cell antibody and drug alone or in combination, for example, antibody-dependent cell-mediated phagocytosis (ADCP), antibody-dependent cell-mediated cytotoxicity (ADCC), complement-mediated cytotoxicity (CDC), and / or inhibition or induction of apoptosis or cell death by a cytotoxic drug.

[1224] As used herein, the term "bystander killing" or "bystander effect" refers to the killing of target-negative cells in the presence of target-positive cells, where killing of target-negative cells is not observed in the absence of target-positive cells. Cell-cell contact or at least proximity between target-positive and target-negative cells enables bystander killing. This type of killing can be distinguished from "off-target killing," which is the indiscriminate killing of target-negative cells. "Off-target killing" can be observed in the absence of target-positive cells.

[1225] In this application, the term "target cell" refers to a cell that expresses an antigen on its surface. The term "target negative" or "target antigen negative" refers to a cell or tissue that lacks expression of the target antigen. The term "target positive" or "target antigen positive" refers to the presence of target antigen expression. For example, a cell or cell line that does not express the target antigen can be described as target negative, while a cell or cell line that expresses the target antigen can be described as target positive.

[1226] As used herein, the term "internalizing" refers to an antibody or antigen-binding fragment that, upon binding to a cell, is able to pass through the cell's lipid bilayer membrane to an internal compartment (i.e., "internalized"), preferably to a degradation compartment within the cell. For example, an internalizing anti-TF antibody is an antibody that is able to enter a cell after binding to TF on the cell membrane.

[1227] In this application, the terms "tissue factor," "TF," "CD142," "tissue factor antigen," "TF antigen," and "CD142 antigen" are used interchangeably herein and, unless otherwise indicated, include any variants, isoforms, and species homologs of human tissue factor naturally expressed by cells or expressed on cells transfected with the tissue factor gene. An "anti-TF antibody" is an antibody as described above that specifically binds to the antigen tissue factor or tissue factor antigen. For example, Tisotumab, and anti-TF antibodies described in patents CN111818943A, CN106938051B, CN110575547A, CN103119065A, WO2011157741, WO 2010066803, and US 9168314 B2. In some embodiments, the anti-TF antibody or antigen-binding fragment thereof is selected from the group consisting of the murine chimeric antibody 1849 with anticoagulant activity in US9920133B2 or the murine chimeric antibody 1859 without anticoagulant activity in US9920133B2.

[1228] In this application, the term "GPC3" refers to any naturally occurring mature GPC3 produced by processing of the GPC3 precursor protein in cells. The term includes GPC3 from any vertebrate, including mammals, such as primates (e.g., humans and cynomolgus monkeys) and rodents (e.g., mice and rats), unless otherwise indicated. The term also includes naturally occurring GPC3 variants, such as splice variants or allelic variants. The heavy chain sequence and light chain amino acid sequence of an exemplary human GPC3 monoclonal antibody can be found in SEQ ID No: 50 and SEQ ID No: 66, respectively, in US Pat. No. 10782300B2.

[1229] In this application, the term "Her2", "human epidermal growth factor receptor 2" or "Her2 / neu" refers to any natural form of human Her2. The term encompasses full-length Her2, as well as any form of human Her2 produced by cell processing (e.g., patent CN103319599). The term also encompasses naturally occurring Her2 variants, including but not limited to splice variants, allelic variants and isoforms. Her2 can be isolated from humans, or can be produced recombinantly or by synthetic methods.

[1230] In this application, the term "optionally linked to other molecular moieties" generally means that the structure is not linked to any other chemical structure, or that the structure is linked to one or more other chemical structures (such as the ligands described in this application) that are different from the structure (for example, linked by a chemical bond or linked by a linker structure).

[1231] The pharmaceutical composition may be in the form of a sterile injectable aqueous or oily suspension for intramuscular and subcutaneous administration. The suspension may be formulated using suitable dispersants or wetting agents and suspending agents as described above according to known techniques. Sterile injectable formulations may also be sterile injectable solutions or suspensions prepared in a nontoxic, parenterally acceptable diluent or solvent, such as a solution prepared in 1,3-butanediol. Furthermore, sterile fixed oils may conveniently be used as a solvent or suspending medium. For example, any blended fixed oil, including synthetic mono- or diglycerides, may be used. Furthermore, fatty acids, such as oleic acid, may also be used to prepare injectable formulations.

[1232] In this application, the term "comprising" generally refers to including the features specified but not excluding other elements. The terms "above" and "below" generally refer to including the number.

[1233] In this application, the term "about" generally refers to a variation within a range of 0.5%-10% above or below the specified value, for example, a variation within a range of 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% above or below the specified value.

[1234] In this application, the term "about" generally means approximately, in the region of, roughly, or around. When the term "about" is used in reference to a numerical range, a cutoff or specific value is used to indicate that the stated value may vary from the recited value by up to 10%. Thus, the term "about" can be used to encompass variations of ±10% or less, ±5% or less, ±1% or less, ±0.5% or less, or ±0.1% or less from the specified value.

[1235] As used herein, a polypeptide, linker fragment, linker or ADC being "responsive to matrix metalloproteinase 2, matrix metalloproteinase 9 or cathepsin B" means that the enzyme can effectively degrade the polypeptide, linker fragment, linker or ADC.

[1236] Detailed Description of the Invention

[1237] polypeptide or its fragment

[1238] In a first aspect, the present application provides a polypeptide P1 or a polypeptide fragment -P1-P2-, wherein the polypeptide P1 and the polypeptide P2 respond to enzymes respectively.

[1239] In some embodiments, P1 comprises a polypeptide responsive to matrix metalloproteinase 2 (MMP-2) or matrix metalloproteinase 9 (MMP-9) or cathepsin B, and P2 comprises a polypeptide responsive to cathepsin B or matrix metalloproteinase 2 (MMP-2) or matrix metalloproteinase 9 (MMP-9).

[1240] In some embodiments, in the -P1-P2-, P1 comprises the following polypeptide structure: -X1-P-X2-X3-Z-, and P2 comprises the following polypeptide structure -X1'-X2'-X3'-X4'-; wherein: P is proline, and X1, X2, X3, Z, X1', X2', X3', and X4' represent absence, a single amino acid, or an amino acid derivative.

[1241] In some embodiments, X1 represents absence, a single amino acid or a single amino acid derivative, X2, X3, and Z represent a single amino acid or a single amino acid derivative, X1' and X2' represent a single amino acid or a single amino acid derivative, and X3' and X4' represent absence or a single amino acid or a single amino acid derivative.

[1242] In some embodiments, the X1 is selected from the group consisting of: absent, Gly (G), Val (V), His (H), Ser (S), Ile (I), the X2 is selected from the group consisting of: Ala (A), Ser (S), Leu (L), Arg (R), Gln (Q), Val (V), Cit, the X3 is selected from the group consisting of: Gly (G), Ser (S), Pro (P), Asn (N), Ala (A), the Z is selected from the group consisting of: Leu (L), Ile (I), Val (V), Phe (F), Met (M), Ala (A), Trp(W), Gln(Q), N,N-dimethyl-3-aminopropionic acid (Dap(Me2)), N,N-dimethyl-4-aminobutyric acid (Dab(Me2)), N,N-dimethylornithine (Orn(Me2)), N,N-dimethyllysine (Lys(Me2)), N,N-diethyl-3-aminopropionic acid (Dap(Et2)), N,N-diethyl-4-aminobutyric acid (Dab(Et2)), N,N-diethylornithine (Orn(Et2)), N,N-diethyllysine (Lys(Et2)).

[1243] For example, polypeptide P1 can be: -SPRGL-, -GPRGL-, -PRGL-, -PRNL- or -GPRNDap(Me2)-.

[1244] In some embodiments, the X1' is selected from the group consisting of Val (V), Als (A), Leu (L), Ile (I), Met (M), Phe (F), Trp (W), Pro (P), Tyr (Y), Gly (G), and Glu (E), and the X2' is selected from the group consisting of Val (V), Leu (L), Ile (I), Met (M), Trp (W), Pro (P), Lys (K), Lys (Me2), Lys (Et2), His (H), A Ala (A), Arg (R), Gly (G), Phe (F), Cit, Orn, Orn (Me2), Orn (Et2), wherein X3' is selected from the group consisting of: absence, Phe (F), Gly (G), Lys (K), Lys (Me2), Lys (Et2), His (H), Ala (A), Arg (R), Cit, Orn, Orn (Me2), Orn (Et2), wherein X4' is selected from the group consisting of: absence, Gly (G), Phe (F).

[1245] In some embodiments, the X1 is selected from: absent, Gly (G), Val (V), His (H), Ser (S), Ile (I), the X2 is selected from: Ala (A), Ser (S), Leu (L), Arg (R), Gln (Q), Val (V), Cit, the X3 is selected from: Gly (G), Ser (S), Pro (P), Asn (N), Ala (A), the Z is selected from: Leu (L), Ile (I), Val (V), Phe (F), M et(M), Ala(A), Trp(W), Gln(Q), N,N-dimethyl-3-aminopropionic acid (Dap(Me2)), N,N-dimethyl-4-aminobutyric acid (Dab(Me2)), N,N-dimethylornithine (Orn(Me2)), N,N-dimethyllysine (Lys(Me2)), N,N-diethyl-3-aminopropionic acid (Dap(Et2)), N,N-diethyl-4-aminobutyric acid (Dab(Et2)), N,N-diethylornithine (Orn(Et 2)), N,N-diethyllysine (Lys(Et2)), wherein X1' is selected from the group consisting of Val(V), Als(A), Leu(L), Ile(I), Met(M), Phe(F), Trp(W), Pro(P), Tyr(Y), Gly(G), Glu(E), and X2' is selected from the group consisting of Val(V), Leu(L), Ile(I), Met(M), Trp(W), Pro(P), Lys(K), Lys(Me2), Lys(Et2), His (H), Ala (A), Arg (R), Gly (G), Phe (F), Cit, Orn, Orn (Me2), Orn (Et2), wherein X3' is selected from the group consisting of: absence, Phe (F), Gly (G), Lys (K), Lys (Me2), Lys (Et2), His (H), Ala (A), Arg (R), Cit, Orn, Orn (Me2), Orn (Et2), and X4' is selected from the group consisting of: absence, Gly (G), Phe (F).

[1246] In some embodiments, P2 is -X1'-X2'-X3'-X4'-, which is further selected from the following fragments: -Val-Cit-, -Ala-Lys-, or -Gly-Gly-Phe-Gly-.

[1247] Wherein, the polypeptide or polypeptide fragment -P1-P2- is selected from the following structures:

[1248] -His-Val-Leu-Asn-Leu-Val-Cit-(SEQ ID NO: 1);

[1249] -Val-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:2);

[1250] -Ile-Pro-Val-Ser-Leu-Val-Cit-(SEQ ID NO:3);

[1251] -Gly-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:4);

[1252] -Gly-Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:5);

[1253] -Gly-Pro-Ala-Gly-Leu-Val-Cit-(SEQ ID NO:6);

[1254] -Gly-Pro-Gln-Gly-Leu-Val-Cit-(SEQ ID NO:7);

[1255] -Gly-Pro-Leu-Gly-Leu-Val-Cit-(SEQ ID NO:8);

[1256] -Gly-Pro-Ser-Gly-Leu-Val-Cit-(SEQ ID NO:9);

[1257] -Gly-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:10);

[1258] -Gly-Pro-Ala-Ala-Leu-Val-Cit-(SEQ ID NO:11);

[1259] -Gly-Pro-Ser-Ala-Leu-Val-Cit-(SEQ ID NO:12);

[1260] -His-Val-Leu-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:13);

[1261] -Val-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:14);

[1262] -Ile-Pro-Val-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:15);

[1263] -Gly-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:16);

[1264] -Gly-Pro-Ala-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:17);

[1265] -Gly-Pro-Ala-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:18);

[1266] -Gly-Pro-Gln-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:19);

[1267] -Gly-Pro-Leu-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:20);

[1268] -Gly-Pro-Ser-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:21);

[1269] -Gly-Pro-Arg-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:22);

[1270] -Gly-Pro-Ala-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:23);

[1271] -Gly-Pro-Ser-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:24);

[1272] -Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:25);

[1273] -Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:26);

[1274] -Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:27);

[1275] -Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:28);

[1276] -Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:29);

[1277] -Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:30);

[1278] -Gly-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:31);

[1279] -Gly-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:32);

[1280] -Gly-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:33);

[1281] -Gly-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:34);

[1282] -Gly-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:35);

[1283] -His-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:36);

[1284] -His-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:37);

[1285] -His-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:38);

[1286] -His-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:39);

[1287] -His-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:40);

[1288] -His-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:41);

[1289] -Ser-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:42);

[1290] -Ser-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:43);

[1291] -Ser-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:44);

[1292] -Ser-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:45);

[1293] -Ser-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:46);

[1294] -Ser-Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:47);

[1295] -Ser-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:48);

[1296] -Gly-Pro-Ala-Pro-Leu-Val-Cit-(SEQ ID NO:49);

[1297] -Gly-Pro-Ala-Asn-Leu-Val-Cit-(SEQ ID NO:50);

[1298] -Gly-Pro-Arg-Gly-Dap(Me2)-Val-Cit-(SEQ ID NO:51);

[1299] Or a variant having one or two mismatches relative to the sequence of any one of SEQ ID NOs: 1-51.

[1300] In the present application, mispairing refers to any mismatch between two sequences, including deletion, increase and substitution of amino acids. For example, the mispairing can be a conservative amino acid substitution. It will be appreciated by those skilled in the art that a conservative amino acid substitution is the substitution of an amino acid with another amino acid having a similar structure or chemical properties (e.g., similar side chains). Exemplary conservative substitutions are described in the art, for example, in Watson et al., Molecular Biology of the Gene, The Bengamin / Cummings Publication Company, 4th edition (1987).

[1301] In some embodiments, a polypeptide or polypeptide fragment -P1-P2- is selected from the following structures:

[1302] -His-Val-Leu-Asn-Leu-Val-Cit-(SEQ ID NO:1);

[1303] -Val-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:2);

[1304] -Ile-Pro-Val-Ser-Leu-Val-Cit-(SEQ ID NO:3);

[1305] -Gly-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:4);

[1306] -Gly-Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:5);

[1307] -Gly-Pro-Ala-Gly-Leu-Val-Cit-(SEQ ID NO:6);

[1308] -Gly-Pro-Gln-Gly-Leu-Val-Cit-(SEQ ID NO:7);

[1309] -Gly-Pro-Leu-Gly-Leu-Val-Cit-(SEQ ID NO:8);

[1310] -Gly-Pro-Ser-Gly-Leu-Val-Cit-(SEQ ID NO:9);

[1311] -Gly-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:10);

[1312] -Gly-Pro-Ala-Ala-Leu-Val-Cit-(SEQ ID NO:11);

[1313] -Gly-Pro-Ser-Ala-Leu-Val-Cit-(SEQ ID NO:12);

[1314] -His-Val-Leu-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:13);

[1315] -Val-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:14);

[1316] -Ile-Pro-Val-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:15);

[1317] -Gly-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:16);

[1318] -Gly-Pro-Ala-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:17);

[1319] -Gly-Pro-Ala-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:18);

[1320] -Gly-Pro-Gln-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:19);

[1321] -Gly-Pro-Leu-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:20);

[1322] -Gly-Pro-Ser-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:21);

[1323] -Gly-Pro-Arg-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:22);

[1324] -Gly-Pro-Ala-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:23);

[1325] -Gly-Pro-Ser-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:24);

[1326] -Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:25);

[1327] -Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:26);

[1328] -Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:27);

[1329] -Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:28);

[1330] -Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:29);

[1331] -Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:30);

[1332] -Gly-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:31);

[1333] -Gly-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:32);

[1334] -Gly-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:33);

[1335] -Gly-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:34);

[1336] -Gly-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:35);

[1337] -His-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:36);

[1338] -His-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:37);

[1339] -His-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:38);

[1340] -His-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:39);

[1341] -His-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:40);

[1342] -His-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:41);

[1343] -Ser-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:42);

[1344] -Ser-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:43);

[1345] -Ser-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:44);

[1346] -Ser-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:45);

[1347] -Ser-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:46);

[1348] -Ser-Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:47);

[1349] -Ser-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:48);

[1350] -Gly-Pro-Ala-Pro-Leu-Val-Cit-(SEQ ID NO:49);

[1351] -Gly-Pro-Ala-Asn-Leu-Val-Cit-(SEQ ID NO:50);

[1352] -Gly-Pro-Arg-Gly-Dap(Me2)-Val-Cit-(SEQ ID NO:51);

[1353] Or a variant having one or two mismatches relative to the sequence of any one of SEQ ID NOs: 1-51.

[1354] linker

[1355] In a second aspect, the present application provides a novel linker structure, wherein the linker comprises a -P1-P2- unit. The -P1-P2- unit is the polypeptide or polypeptide fragment described in the first aspect. The linker has the following general formula: C1-La-L1-P1-P2-L2:

[1356] Wherein C1 is a linker, connecting the biological ligand units;

[1357] La is empty or a tuning spacer;

[1358] L1 is empty or a spacer;

[1359] P1-P2 are the polypeptides or polypeptide fragments described in the first aspect;

[1360] L2 is empty or a spacer, connecting a payload with a specific biological function;

[1361] The positions of La and L1 can be swapped.

[1362] In some embodiments, the biological ligand unit comprises an antibody, an antibody fragment, a protein, a polypeptide, a polynucleotide, or a small chemical molecule.

[1363] In some embodiments, linker C1 is selected from the following structures:

[1364] in Indicates the connection position with the biological ligand unit, Indicates the connection position with La.

[1365] In some embodiments, the C1 comprises the following structure: -(succinimid-3-yl-N)-, -CH2-C(=O)-, or -C(=O)-.

[1366] In some embodiments, the La may be empty.

[1367] In some embodiments, the La is selected from the following structures or a combination of the following structures:

[1368] R1-SS-R2,

[1369] R1-(CH2CH2O) m -R2,

[1370] R1-(CH2CH2O) m -CH2CH2S-(CH2CH2O) n -R2,

[1371] R1-(CH2CH2O) m -CH2CH2NH-(CH2CH2O) n -R2,

[1372] R1-(CH2CH2O) m -CH2CH2(C=O)NH-(CH2CH2O) n -R2,

[1373] a is 0 or 1;

[1374] m is any integer from 1 to 20;

[1375] n is any integer from 1 to 20;

[1376] R3 is H, -C1-C10 alkylene-, -C3-C8 carbocyclyl-, -C2-C10 heteroaryl-, -C3-C24 alkylheteroaryl-, wherein -C1-C10 alkylene-, -C3-C8 carbocyclyl-, -C2-C10 heteroaryl-, -C3-C24 alkylheteroaryl- may be substituted by one or more heteroatoms, which may be O, S, NR 3 , where R 3 is H or -C1-C4 alkylene,

[1377] R1 is a connecting structure with C1, and R2 is a connecting structure with L1.

[1378] In some embodiments, R1 is a single bond or is selected from the following structures:

[1379] -X 1 -C1-C10 alkylene-X 2 -、-X 1 -C1-C10 heteroalkylene-X 2 -、-X 1 -C3-C8 carbocyclyl-X 2 -、-X 1 -Arylene-X 2 -、-X 1 -C1-C10 alkylene-arylene-X 2 -、-X 1 -Arylene-C1-C10 alkylene-X 2 -、-X 1 -C1-C10 alkylene-(C3-C8 carbocyclyl)-X 2 -、-X 1 -(C3-C8 carbocyclyl)-C1-C10 alkylene-X 2 -、-X 1 -C3-C8 heterocyclyl-X 2 -、-X 1 -C1-C10 alkylene-(C3-C8 heterocyclyl)-X 2 -、-X 1 -(C3-C8 heterocyclyl)-C1-C10 alkylene-X 2 -;

[1380] wherein X 1 is absent or selected from O, S, NH, C(=O), NHC(=O), C(=O)NH, NHC(=O)NH, X 2is absent or selected from O, NH, S, C(═O), C(═O)NH, NHC(═O), and NHC(═O)NH.

[1381] Wherein said R1 when not a single bond, is optionally substituted by a basic unit (BU), said basic unit being -(CH2) x NH2, -(CH2) x NHR m or -(CH2) x N(R m )2; wherein x is any integer from 1 to 4; and each R m are independently selected from C1-C6 alkyl and C1-C6 haloalkyl, or two R m The groups combine with the nitrogen to which they are attached to form a 4- to 6-membered heterocycloalkyl ring, or an azetidinyl, pyrrolidinyl, or piperidinyl group.

[1382] In some embodiments, R2 is a single bond or is selected from the following structures:

[1383] -X 3 -C1-C10 alkylene-X 4 -、-X 3 -C1-C10 heteroalkylene-X 4 -、-X 3 -C3-C8 carbocyclyl-X 4 -、-X 3 -Arylene-X 4 -、-X 3 -C1-C10 alkylene-arylene-X 4 -、-X 3 -Arylene-C1-C10 alkylene-X 4 -、-X 3 -C1-C10 alkylene-(C3-C8 carbocyclyl)-X 4 -、-X 3 -(C3-C8 carbocyclyl)-C1-C10 alkylene-X 4 -、-X 3 -C3-C8 heterocyclyl-X 4 -、-X 3 -C1-C10 alkylene-(C3-C8 heterocyclyl)-X 4 -、-X 3 -(C3-C8 heterocyclyl)-C1-C10 alkylene-X 4 -;

[1384] wherein X 3 is absent or O, S, NH, C(=O), NHC(=O), C(=O)NH, NHC(=O)NH, X4 is absent or selected from O, NH, S, C(═O), C(═O)NH, NHC(═O), and NHC(═O)NH.

[1385] Wherein said R2 when not a single bond, is optionally substituted by a basic unit (BU), said basic unit being -(CH2) x NH2, -(CH2) x NHR m or -(CH2) x N(R m )2; wherein x is any integer from 1 to 4; and each R m are independently selected from C1-C6 alkyl and C1-C6 haloalkyl, or two R m The groups combine with the nitrogen to which they are attached to form a 4- to 6-membered heterocycloalkyl ring, or an azetidinyl, pyrrolidinyl, or piperidinyl group.

[1386] For example, La may be the following structures: -CH2CH2C(O)NH-, -OC(O)-, -OC(O)-(CH2CH2O)4-.

[1387] In certain embodiments, L1 is selected from the following structures:

[1388] -X 5 -C1-C10 alkylene-X 6 -、-X 5 -C1-C10 heteroalkylene-X 6 -、-X 5 -C3-C8 carbocyclyl-X 6 -、-X 5 -Arylene-X 6 -、-X 5 -heteroarylene-X 6 -、-X 5 -C1-C10 alkylene-arylene-X 6 -、-X 5 -C1-C10 alkylene-heteroarylene-X 6 -、-X 5 -Arylene-C1-C10 alkylene-X 6 -、-X 5 -heteroarylene-C1-C10alkylene-X 6 -、-X 5 -C1-C10 alkylene-(C3-C8 carbocyclyl)-X 6 -、-X 5 -(C3-C8 carbocyclyl)-C1-C10 alkylene-X 6 -、-X 5-C3-C8 heterocyclyl-X 6 -、-X 5 -C1-C10 alkylene-(C3-C8 heterocyclyl)-X 6 -、-X 5 -(C3-C8 heterocyclyl)-C1-C10 alkylene-X 6 -、-X 5 -C3-C8 heterocyclyl-arylene-X 6 -、-X 5 -Arylene-C3-C8 heterocyclyl-X 6 -、-X 5 -C3-C8 heterocyclyl-heteroarylene-X 6 -、-X 5 -heteroarylene-C3-C8heterocyclyl-X 6 -;

[1389] wherein X 5 is absent or selected from O, S, NH, C(=O), NHC(=O), C(=O)NH, NHC(=O)NH, X 6 is absent or selected from O, S, NH, C(=O), NHC(=O), C(=O)NH, NHC(=O)NH;

[1390] Wherein, any methylene unit of L1 can be independently replaced by -O-, -S-, -N(R n )-、-CH(N(R n )2)-、-CH(P(R n )2)-、-C(N(R n )2)2-、-C(OR n )(N(R n )2)-, -C(=O)-, -OC(=O)-, -C(=O)O-, -N(R n )C(=O)-、-C(=O)N(R n )-、-SO-、-SO2-、-N(R n )SO2-、-SO2N(R n )-、-P(R n )-、-P(=O)(R n )-, -P(=O)(O)-, -C(=S)-, -C(=NR n )-、-SS-、-N=N-、-N(R n )-N=、=NN(R n )-, -C=N- or -N=C-; wherein each R nare independently selected from H, C1-C6 alkyl, C3-C8 carbocyclyl, C1-C6 haloalkyl, or two R n The groups combine with the nitrogen or phosphorus to which they are attached to form a 4- to 6-membered heterocycloalkyl ring, or an azetidinyl, pyrrolidinyl, or piperidinyl group.

[1391] In some embodiments, L2 is empty.

[1392] In some embodiments, L2 is selected from the following structures:

[1393] in Indicates the connection position with the polypeptide or polypeptide fragment -P1-P2-, Indicates the location of the payload connection with a specific biological function;

[1394] m is any integer from 1 to 20;

[1395] n is any integer from 1 to 6;

[1396] R b Independently selected from H, C1-C6 alkyl, C3-C8 carbocyclyl, C1-C6 haloalkyl, C3-C8 heterocyclyl, C1-C6 alkyl-C3-C8 heterocyclyl, substituted aryl, substituted heteroaryl, C1-C6 alkyl-substituted aryl, C1-C6 alkyl-substituted heteroaryl.

[1397] Wherein any methylene unit of L2 can be independently replaced by -CHX-, -C(X2)-, -C3-C8 carbocyclyl-, -C3-C8 heterocyclyl-, -arylene-, -heteroarylene-, -O-, -S-, -N(R n )-、-CH(N(R n )2)-、-CH(P(R n )2)-、-C(N(R n )2)2-、-C(OR n )(N(R n )2)-, -C(=O)-, -OC(=O)-, -C(=O)O-, -N(R n )C(=O)-、-C(=O)N(R n )-、-SO-、-SO2-、-N(R n )SO2-、-SO2N(R n )-、-P(R n )-、-P(=O)(R n )-, -P(=O)(O)-, -C(=S)-, -C(=NR n )-、-SS-、-N=N-、-N(R n)-N=、=NN(R n )-, -C=N- or -N=C-; wherein X represents halogen or deuterium, each R n are independently selected from H, C1-C6 alkyl, C3-C8 carbocyclyl, C1-C6 haloalkyl, or two R n The groups combine with the nitrogen or phosphorus to which they are attached to form a 4- to 6-membered heterocycloalkyl ring, or an azetidinyl, pyrrolidinyl, or piperidinyl group.

[1398] In some embodiments, L2 may be empty.

[1399] In some embodiments, L2 is selected from the following structures:

[1400] in Indicates the connection position with the polypeptide or polypeptide fragment -P1-P2-, Indicates the location of the payload connection with a specific biological function;

[1401] W and Y are each independently N or CR Y , and each R Y are independently H or C1-C10 alkyl;

[1402] m is any integer from 0 to 20;

[1403] X is halogen or deuterium.

[1404] In some embodiments, L2 may be:

[1405] In some embodiments, the linker is selected from the following structures:

[1406] -(Succinimidyl-3-yl-N)-CH2-C(=O)-P1-P2-;

[1407] -(Succinimidyl-3-yl-N)-CH2CH2-C(=O)-P1-P2-;

[1408] -(succinimidyl-3-yl-N)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-;

[1409] -(Succinimidyl-3-yl-N)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-;

[1410] -(Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-;

[1411] -(Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-;

[1412] -(Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-;

[1413] -(Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-;

[1414] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-;

[1415] -(Succinimidyl-3-yl-N)-CH2-C(=O)-P1-P2-PAB-;

[1416] -(Succinimidyl-3-yl-N)-CH2CH2-C(=O)-P1-P2-PAB-;

[1417] -(Succinimidyl-3-yl-N)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-;

[1418] -(Succinimidyl-3-yl-N)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-;

[1419] -(Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-;

[1420] -(Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-;

[1421] -(Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-;

[1422] -(Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-;

[1423] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-PAB-;

[1424] in:

[1425] Indicates the connection position with the biological ligand unit;

[1426] PAB represents the following structure:

[1427] The -P1-P2- represents the following structure:

[1428] -His-Val-Leu-Asn-Leu-Val-Cit-(SEQ ID NO: 1);

[1429] -Val-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:2);

[1430] -Ile-Pro-Val-Ser-Leu-Val-Cit-(SEQ ID NO:3);

[1431] -Gly-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:4);

[1432] -Gly-Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:5);

[1433] -Gly-Pro-Ala-Gly-Leu-Val-Cit-(SEQ ID NO: 6);

[1434] -Gly-Pro-Gln-Gly-Leu-Val-Cit-(SEQ ID NO:7);

[1435] -Gly-Pro-Leu-Gly-Leu-Val-Cit-(SEQ ID NO:8);

[1436] -Gly-Pro-Ser-Gly-Leu-Val-Cit-(SEQ ID NO:9);

[1437] -Gly-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO: 10);

[1438] -Gly-Pro-Ala-Ala-Leu-Val-Cit-(SEQ ID NO: 11);

[1439] -Gly-Pro-Ser-Ala-Leu-Val-Cit-(SEQ ID NO:12);

[1440] -His-Val-Leu-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:13);

[1441] -Val-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:14);

[1442] -Ile-Pro-Val-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:15);

[1443] -Gly-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:16);

[1444] -Gly-Pro-Ala-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:17);

[1445] -Gly-Pro-Ala-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:18);

[1446] -Gly-Pro-Gln-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:19);

[1447] -Gly-Pro-Leu-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:20);

[1448] -Gly-Pro-Ser-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:21);

[1449] -Gly-Pro-Arg-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:22);

[1450] -Gly-Pro-Ala-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:23);

[1451] -Gly-Pro-Ser-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:24);

[1452] -Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:25);

[1453] -Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:26);

[1454] -Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:27);

[1455] -Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:28);

[1456] -Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:29);

[1457] -Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:30);

[1458] -Gly-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:31);

[1459] -Gly-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:32);

[1460] -Gly-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:33);

[1461] -Gly-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:34);

[1462] -Gly-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:35);

[1463] -His-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:36);

[1464] -His-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:37);

[1465] -His-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:38);

[1466] -His-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:39);

[1467] -His-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:40);

[1468] -His-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:41);

[1469] -Ser-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:42);

[1470] -Ser-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:43);

[1471] -Ser-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:44);

[1472] -Ser-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:45);

[1473] -Ser-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:46);

[1474] -Ser-Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:47);

[1475] -Ser-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:48);

[1476] -Gly-Pro-Ala-Pro-Leu-Val-Cit-(SEQ ID NO:49);

[1477] -Gly-Pro-Ala-Asn-Leu-Val-Cit-(SEQ ID NO:50);

[1478] -Gly-Pro-Arg-Gly-Dap(Me2)-Val-Cit-(SEQ ID NO:51);

[1479] Alternatively, a variant having one or two mismatches relative to the sequence of any one of SEQ ID NOs: 1-51.

[1480] In some embodiments, the linker can be selected from:

[1481] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLGLGGFG-;

[1482] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLGGFG-;

[1483] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAALGGFG-;

[1484] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-;

[1485] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPASLVCit-PAB-;

[1486] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLVCit-PAB-;

[1487] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPSALVCit-PAB-;

[1488] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-PRNLVCit-PAB-;

[1489] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRNLVCit-PAB-;

[1490] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGDap(Me2)VCit-PAB-;

[1491] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-SPRNLVCit-PAB-;

[1492] -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLSLVCit-PAB-;

[1493] in:

[1494] PAB represents the following structure:

[1495] Linker-payload

[1496] In a third aspect, the present application provides a linker drug comprising the linker described in the second aspect.

[1497] In some embodiments, the linker drug has the following structure:

[1498] C1'—La—L1—P1—P2—L2-D

[1499] Wherein, the C1' is a functional group that reacts with a biological ligand;

[1500] La is empty or a tuning spacer;

[1501] L1 is empty or a spacer;

[1502] P1-P2 are the polypeptides or polypeptide fragments described in the first aspect;

[1503] L2 is empty or a spacer, connecting a payload with a specific biological function;

[1504] D is the drug unit;

[1505] The positions of La and L1 can be swapped.

[1506] It should be noted that biological ligands include antibodies, antibody fragments, proteins, peptides, polynucleotides, and small chemical molecules, and the functional group C1' can react with the upper functional group of the biological ligand to connect the linker drug to the biological ligand. For example, C1' is When the linker is used, it can react with the side chain -SH of the monoclonal antibody Cys reduced by a reducing agent, thereby connecting the linker drug to the antibody.

[1507] In some embodiments, C1' is selected from functional groups reactive with amino, sulfhydryl, keto, azide, and alkyne groups.

[1508] In some embodiments, C1' is selected from a substrate of an enzyme-catalyzed reaction. For example, Gly-Gly-Gly- can be recognized by the Sortase A enzyme.

[1509] In some embodiments, C1' is selected from the following structures:

[1510] in:

[1511] R X is selected from halogen, -SPh. S Selected from halogen, sulfone group, tertiary amine salt group, diazonium salt group, -OMs, MeSO2-, and CF3SO3-.

[1512] Indicates the connection position with La.

[1513] In this disclosure, the drug unit refers to a portion (fragment or group) of an antibody-drug conjugate (or antibody-drug conjugate, ADC) known in the art, which can form a biologically active drug (such as a small molecule cytotoxic drug, including a group after losing an atom or atomic group) or its derivative (such as a precursor) after the linker is cleaved / degraded / enzymatically cleaved between tumor tissues or within tumor cells. For the avoidance of doubt, "drug" does not only refer to "drugs" approved by medical regulatory authorities, but also includes any molecule with potential therapeutic biological activity in clinical practice, or in R&D and academic research.

[1514] In some embodiments, the drug unit D is selected from an immunomodulator, a protein degrader, and a cytotoxic agent.

[1515] In some embodiments, the Drug unit D is selected from a cytotoxic agent.

[1516] In some embodiments, the drug unit comprises: amanitins, anthracyclines, auristatins, baccatins, calicheamicins, camptothecins, cemadotins, colchicines, colcimids, combretastatins, cryptophycins, discodermolides, duocarmycins, docetaxel, doxorubicin, duocarmycins, echinomycins, eleutherobins, epothilones, estramustines, lexitropsins, maytansines, maytansinoids, methotrexate, netropsins, pyrrolo[2,1-c][1,4]benzodi-azepines (PBDs), puromycins, rhizoxins, SN-38, taxanes, tubulysins, vincaalkaloids, tetrahydroisoquinoline alkaloids or derivatives thereof, protein degraders or derivatives thereof.

[1517] In some embodiments, the drug unit comprises a DNA topoisomerase I inhibitor, a tubulin inhibitor, a DNA replication inhibitor, or a protein degrader.

[1518] In some embodiments, the drug unit comprises maytansine alkaloid or its derivatives, camptothecin or its derivatives, auristatin or its derivatives, tetrahydroisoquinoline alkaloid or its derivatives, BTK protein degrader and its derivatives, GSPT1 protein degrader and its derivatives.

[1519] In some embodiments, the drug unit comprises DM1, DM4, DX8951, MMAE, Dxd, SN38, Trabectedin (ET743), Lurbinectedin, CC885, CC-90009.

[1520] In some embodiments, the linker drug has the following structure:

[1521] (Maleimido-N-yl)-CH2-C(=O)-P1-P2-D;

[1522] (Maleimido-N-yl)-CH2CH2-C(=O)-P1-P2-D;

[1523] (Maleimido-N-yl)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D;

[1524] (Maleimide-N-yl)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D;

[1525] (Maleimido-N-yl)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D;

[1526] (Maleimido-N-yl)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D;

[1527] (Maleimido-N-yl)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D;

[1528] (Maleimido-N-yl)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D;

[1529] (Maleimido-N-yl)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-D;

[1530] (Maleimido-N-yl)-CH2-C(=O)-P1-P2-PAB-D;

[1531] (Maleimido-N-yl)-CH2CH2-C(=O)-P1-P2-PAB-D;

[1532] (Maleimido-N-yl)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D;

[1533] (Maleimide-N-yl)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D;

[1534] (Maleimido-N-yl)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D;

[1535] (Maleimido-N-yl)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D;

[1536] (Maleimido-N-yl)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D;

[1537] (Maleimido-N-yl)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D;

[1538] (Maleimido-N-yl)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-PAB-D;

[1539] (N-hydroxysuccinimide ester-1-yl)-CH2-C(=O)-P1-P2-D;

[1540] (N-hydroxysuccinimide ester-1-yl)-CH2CH2-C(=O)-P1-P2-D;

[1541] (N-hydroxysuccinimide ester-1-yl)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D;

[1542] (N-hydroxysuccinimide ester-1-yl)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D;

[1543] (N-hydroxysuccinimide ester-1-yl)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D;

[1544] (N-hydroxysuccinimide ester-1-yl)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D;

[1545] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2-C(=O)-P1-P2-D;

[1546] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D;

[1547] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D;

[1548] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2C(=O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D;

[1549] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2C(=O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D;

[1550] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2O-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D;

[1551] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2O-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D;

[1552] (N-hydroxysuccinimide ester-1-yl)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-D;

[1553] (N-hydroxysuccinimide ester-1-yl)-CH2-C(=O)-P1-P2-PAB-D;

[1554] (N-hydroxysuccinimide ester-1-yl)-CH2CH2-C(=O)-P1-P2-PAB-D;

[1555] (N-hydroxysuccinimide ester-1-yl)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D;

[1556] (N-hydroxysuccinimide ester-1-yl)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D;

[1557] (N-hydroxysuccinimide ester-1-yl)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D;

[1558] (N-hydroxysuccinimide ester-1-yl)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D;

[1559] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2-C(=O)-P1-P2-PAB-D;

[1560] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D;

[1561] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D;

[1562] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2C(=O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D;

[1563] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2C(=O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D;

[1564] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2O-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D;

[1565] (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2O-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D;

[1566] (N-hydroxysuccinimide ester-1-yl)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-PAB-D;

[1567] Gly-Gly-Gly-NH-CH2CH2CH2CH2CH2-C(=O)-P1-P2-D;

[1568] Gly-Gly-Gly-NH-CH2CH2CH2CH2CH2-C(=O)-P1-P2-PAB-D;

[1569] The (N-hydroxysuccinimide ester-1-yl)- has the following structure:

[1570] in Indicates the position of connection with the linker;

[1571] The -P1-P2- represents the following structure:

[1572] -His-Val-Leu-Asn-Leu-Val-Cit-(SEQ ID NO: 1);

[1573] -Val-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:2);

[1574] -Ile-Pro-Val-Ser-Leu-Val-Cit-(SEQ ID NO:3);

[1575] -Gly-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:4);

[1576] -Gly-Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:5);

[1577] -Gly-Pro-Ala-Gly-Leu-Val-Cit-(SEQ ID NO: 6);

[1578] -Gly-Pro-Gln-Gly-Leu-Val-Cit-(SEQ ID NO:7);

[1579] -Gly-Pro-Leu-Gly-Leu-Val-Cit-(SEQ ID NO:8);

[1580] -Gly-Pro-Ser-Gly-Leu-Val-Cit-(SEQ ID NO:9);

[1581] -Gly-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO: 10);

[1582] -Gly-Pro-Ala-Ala-Leu-Val-Cit-(SEQ ID NO: 11);

[1583] -Gly-Pro-Ser-Ala-Leu-Val-Cit-(SEQ ID NO:12);

[1584] -His-Val-Leu-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:13);

[1585] -Val-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:14);

[1586] -Ile-Pro-Val-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:15);

[1587] -Gly-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:16);

[1588] -Gly-Pro-Ala-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:17);

[1589] -Gly-Pro-Ala-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:18);

[1590] -Gly-Pro-Gln-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:19);

[1591] -Gly-Pro-Leu-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:20);

[1592] -Gly-Pro-Ser-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:21);

[1593] -Gly-Pro-Arg-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:22);

[1594] -Gly-Pro-Ala-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:23);

[1595] -Gly-Pro-Ser-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:24);

[1596] -Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:25);

[1597] -Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:26);

[1598] -Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:27);

[1599] -Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:28);

[1600] -Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:29);

[1601] -Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:30);

[1602] -Gly-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:31);

[1603] -Gly-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:32);

[1604] -Gly-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:33);

[1605] -Gly-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:34);

[1606] -Gly-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:35);

[1607] -His-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:36);

[1608] -His-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:37);

[1609] -His-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:38);

[1610] -His-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:39);

[1611] -His-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:40);

[1612] -His-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:41);

[1613] -Ser-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:42);

[1614] -Ser-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:43);

[1615] -Ser-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:44);

[1616] -Ser-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:45);

[1617] -Ser-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:46);

[1618] -Ser-Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:47);

[1619] -Ser-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:48);

[1620] -Gly-Pro-Ala-Pro-Leu-Val-Cit-(SEQ ID NO:49);

[1621] -Gly-Pro-Ala-Asn-Leu-Val-Cit-(SEQ ID NO:50);

[1622] -Gly-Pro-Arg-Gly-Dap(Me2)-Val-Cit-(SEQ ID NO:51);

[1623] The D is camptothecin or its derivatives, auristatin or its derivatives, tetrahydroisoquinoline alkaloids or its derivatives, PROTAC protein degraders, molecular glue protein degraders, preferably DX8951, MMAE, Dxd, SN38, ET743, CC885, CC-90009.

[1624] In certain embodiments, the linker drug has the following structure:

[1625] MC-GPAGLVCit-PAB-DX8951;

[1626] MC-GPLGLGGFG-DX8951;

[1627] MC-GPASLVCit-PAB-DX8951;

[1628] MC-GPRGLVCit-PAB-DX8951;

[1629] MC-GPSALVCit-PAB-DX8951;

[1630] MC-GPRGLGGFG-DX8951;

[1631] MC-GPAALGGFG-DX8951;

[1632] MC-PRNLVCit-PAB-DX8951;

[1633] MC-GPRNLVCit-PAB-DX8951;

[1634] MC-GPRGDap(Me2)LVCit-PAB-DX8951;

[1635] MC-SPRNLVCit-PAB-DX8951;

[1636] MC-GPLSLVCit-PAB-DX8951;

[1637] MC-PRGLVCit-PAB-DX8951;

[1638] MC-PRPLVCit-PAB-DX8951;

[1639] MC-GPRSLVCit-PAB-DX8951;

[1640] MC-GPRPLVCit-PAB-DX8951;

[1641] MC-GPCitPLVCit-PAB-DX8951;

[1642] MC-HPRGLVCit-PAB-DX8951;

[1643] MC-HPCitPLVCit-PAB-DX8951;

[1644] MC-SPCitGLVCit-PAB-DX8951;

[1645] MC-GPAPLVCit-PAB-DX8951;

[1646] MC-PLSLVCit-PAB-DX8951;

[1647] MC-SPCitSLVCit-PAB-DX8951;

[1648] MC-PRSLVCit-PAB-DX8951;

[1649] MC-SPRSLVCit-PAB-DX8951;

[1650] MC-PCitSLVCit-PAB-DX8951;

[1651] MC-PCitPLVCit-PAB-DX8951;

[1652] MC-PCitGLVCit-PAB-DX8951;

[1653] MC-GPCitGLVCit-PAB-DX8951;

[1654] MC-SPRGLVCit-PAB-DX8951;

[1655] MC-SPRPLVCit-PAB-DX8951;

[1656] MC-SPCitPLVCit-PAB-DX8951;

[1657] MC-GPANLVCit-PAB-DX8951

[1658] MC-GPCitNLVCit-PAB-DX8951;

[1659] MC-SPCitSDap(Me2)VCit-PAB-DX8951;

[1660] MC-PRSDap(Me2)VCit-PAB-DX8951;

[1661] MC-GPAGLVCit-PAB-MMAE;

[1662] MC-GPRGLVCit-PAB-MMAE;

[1663] MC-PRNLVCit-PAB-MMAE;

[1664] MC-GPRNLVCit-PAB-MMAE;

[1665] MC-GPRGDap(Me2)LVCit-PAB-MMAE;

[1666] MC-SPRNLVCit-PAB-MMAE;

[1667] MC-GPLSLVCit-PAB-MMAE;

[1668] MC-SPCitSLVCit-PAB-MMAE

[1669] MC-SPCitSDap(Me2)VCit-PAB-MMAE

[1670] MC-SPCitSDap(Me2)VCit-PAB-MMAE

[1671] MC-GPAGLVCit-PAB-ET743;

[1672] MC-GPRNLVCit-PAB-ET743;

[1673] MC-SPCitSLVCit-PAB-ET743;

[1674] The MC- has the following structure:

[1675] in Indicates the position of connection with the polypeptide

[1676] The DX8951 has the following structure:

[1677] in Indicates the connection location of DX8951;

[1678] The MMAE has the following structure:

[1679] in Indicates the connection position of MMAE.

[1680] The ET-743 has the following structure:

[1681] in Indicates the connection location of ET743.

[1682] Ab+Linker+Payload Antibody-Drug Conjugate

[1683] In a fourth aspect, the present application discloses a conjugate comprising polypeptide or polypeptide fragment-P1-P2-units.

[1684] In some embodiments, the conjugate has the general formula:

[1685] in:

[1686] Bp is the biological ligand;

[1687] L is a linker comprising a -P1-P2- unit, wherein the -P1-P2- unit is the polypeptide or polypeptide fragment described in the first two aspects;

[1688] D is the drug unit;

[1689] p is any integer between 1 and 20.

[1690] In some embodiments, the conjugate has the general formula:

[1691] in:

[1692] Bp is the biological ligand;

[1693] C1, La, L1, and L2 are defined as described in the first two aspects;

[1694] -P1-P2- unit is the polypeptide or polypeptide fragment described in the first two aspects;

[1695] D is the drug unit;

[1696] p is any integer between 1 and 20.

[1697] In some embodiments, the biological ligand targets a cell surface receptor or a tumor associated antigen.

[1698] In some embodiments, the cell surface receptors or tumor-associated antigens include: EGFR, HER2, HER3, c-Met, claudin18.2, TROP-2, folate receptor alpha (FRα), ROR1, ROR2, BCMA, PSMA, CD19, CD20, CD22, CD30, CD33, CD37, CD46, CD48, CD56, CD79b, CD123, CD138, CD166, CD276, CEACAM5, SDC1, CD74, CD70, MUC1, MUC16, GUCY2C, MSLN, SCL34A2, FOLH1, TPBC, PVRL4, STEAP1, SCL44A4, NACM1, EDNRB, GPNMB, FGFR, SEZ6, Nectin-4, PD-L1, Tissue Factor(TF), B7-H3, B7-H4, LY6E, LIV-1, CDH4, CDH6, ITGB6, GPC1, GPC3, ENPP3, KAAG1, FZD7, SSTR2, DLK1, T MEFF1, TM4SF1, SLAMF7, DLL3, FLT3, TNFRSF10B, EPCAM, AXL, IL2RA, 5T4, FAP, ICAM-1, IGF-1R, TNFR1, GLP-1.

[1699] In some embodiments, the biological ligand includes an antibody or an antigen-binding fragment thereof, a modified antibody or an antigen-binding fragment thereof, an oligonucleotide, a DNA fragment, an aptamer, a polypeptide, a nanoparticle with targeting properties, or a chemically synthesized small molecule.

[1700] In some embodiments, the biological ligand comprises an antibody or antigen-binding fragment thereof, or a modified antibody or antigen-binding fragment thereof.

[1701] In some embodiments, the antibody comprises a monoclonal antibody, a polyclonal antibody, a dimer, a multimer, a multispecific antibody, a whole antibody, an antibody fragment, a human antibody, a humanized antibody, or a chimeric antibody.

[1702] In some embodiments, the modified antibodies include modified monoclonal antibodies, modified polyclonal antibodies, modified dimers, modified multimers, modified multispecific antibodies, modified whole antibodies, modified antibody fragments, modified human antibodies, modified humanized antibodies, or modified chimeric antibodies.

[1703] In some embodiments, the antigen binding fragment comprises a Fab, a Fab', an Fv fragment, a F(ab')2, a scFv, a di-scFv and / or a dAb.

[1704] In some embodiments, the modified antigen-binding fragment comprises a modified Fab, a modified Fab', a modified Fv fragment, a modified F(ab')2, a modified scFv, a modified di-scFv and / or a modified dAb.

[1705] In some embodiments, the antibody is a monoclonal antibody or a modified monoclonal antibody.

[1706] In some embodiments, the antibody is a human antibody, a humanized antibody, or a chimeric antibody, wherein the modified antibody is a modified human antibody, a modified humanized antibody, or a modified chimeric antibody.

[1707] In some embodiments, the modification methods include: amino acid point mutation, glycosylation modification, and polypeptide modification.

[1708] In some embodiments, the antibody or antigen-binding fragment thereof, modified antibody or modified antigen-binding fragment thereof is: anti-EGFR, anti-HER2, anti-HER3, anti-c-Met, anti-claudin18.2, anti-TROP-2, anti-FRα, anti-ROR1, anti-ROR2, anti-BCMA, anti-PSMA, anti-CD19, anti-CD20, anti-CD22, anti-CD30, anti-CD33, anti- CD37, anti-CD46, anti-CD48, anti-CD56, anti-CD79b, anti-CD123, anti-CD138, anti-CD166, anti-CD276, anti-CEACAM5, anti -SDC1, anti-CD74, anti-CD70, anti-MUC1, anti-MUC16, anti-GUCY2C, anti-MSLN, anti-SCL34A2, anti-FOLH1, anti-TPBC, anti- PVRL4, anti-STEAP1, anti-SCL44A4, anti-NACM1, anti-EDNRB, anti-GPNMB, anti-FGFR, anti-SEZ6, anti-Nectin-4, anti-PD-L 1. anti-TF, anti-B7-H3, anti-B7-H4, anti-LY6E, anti-LIV-1, anti-CDH4, anti-CDH6, anti-ITGB6, anti-GPC1, anti-GPC3, anti -ENPP3, anti-KAAG1, anti-FZD7, anti-SSTR2, anti-DLK1, anti-TMEFF1, anti-TM4SF1, anti-SLAMF7, anti-DLL3, anti-FLT3, an ti-TNFRSF10B, anti-EPCAM, anti-AXL, anti-IL2RA, anti-5T4, anti-FAP, anti-ICAM-1, anti-IGF-1R, anti-TNFR1, anti-GLP-1.

[1709] In some embodiments, the antibody or antigen-binding fragment thereof, modified antibody or modified antigen-binding fragment thereof is: anti-Her2, anti-TF, anti-ROR1, anti-GPC3 antibody or antigenic structural fragment thereof.

[1710] In some embodiments, the antibody is: Trastuzumab, Tisotumab, Zilovertamab, Codrituzumab.

[1711] In some embodiments, the conjugate is an antibody drug conjugate, wherein the antibody drug conjugate is selected from the following structures:

[1712] Ab-((Succinimidyl-3-yl-N)-CH2-C(=O)-P1-P2-D) n ;

[1713] Ab-((Succinimidyl-3-yl-N)-CH2CH2-C(=O)-P1-P2-D) n ;

[1714] Ab-((Succinimidyl-3-yl-N)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D) n ;

[1715] Ab-((Succinimidyl-3-yl-N)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D) n ;

[1716] Ab-((Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D) n ;

[1717] Ab-((Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D) n ;

[1718] Ab-((Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D) n ;

[1719] Ab-((Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D) n ;

[1720] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-D) n ;

[1721] Ab-((Succinimidyl-3-yl-N)-CH2-C(=O)-P1-P2-PAB-D) n ;

[1722] Ab-((Succinimidyl-3-yl-N)-CH2CH2-C(=O)-P1-P2-PAB-D) n ;

[1723] Ab-((Succinimidyl-3-yl-N)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D) n ;

[1724] Ab-((Succinimidyl-3-yl-N)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D) n ;

[1725] Ab-((Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D) n ;

[1726] Ab-((Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D) n ;

[1727] Ab-((Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D) n ;

[1728] Ab-((Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D) n ;

[1729] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-PAB-D) n ;

[1730] The Ab represents a monoclonal antibody or an antigenic structural fragment thereof, or a modified monoclonal antibody or an antigen-binding fragment thereof, preferably an anti-Her2, anti-TF, anti-ROR1, anti-GPC3 antibody or an antigenic structural fragment thereof;

[1731] The -P1-P2- represents the following structures:

[1732] -His-Val-Leu-Asn-Leu-Val-Cit- (SEQ ID NO:1);

[1733] -Val-Pro-Leu-Ser-Leu-Val-Cit- (SEQ ID NO:2);

[1734] -Ile-Pro-Val-Ser-Leu-Val-Cit- (SEQ ID NO:3);

[1735] -Gly-Pro-Leu-Ser-Leu-Val-Cit- (SEQ ID NO:4);

[1736] -Gly-Pro-Ala-Ser-Leu-Val-Cit- (SEQ ID NO:5);

[1737] -Gly-Pro-Ala-Gly-Leu-Val-Cit- (SEQ ID NO:6);

[1738] -Gly-Pro-Gln-Gly-Leu-Val-Cit- (SEQ ID NO:7);

[1739] -Gly-Pro-Leu-Gly-Leu-Val-Cit- (SEQ ID NO:8);

[1740] -Gly-Pro-Ser-Gly-Leu-Val-Cit- (SEQ ID NO:9);

[1741] -Gly-Pro-Arg-Gly-Leu-Val-Cit- (SEQ ID NO:10);

[1742] -Gly-Pro-Ala-Ala-Leu-Val-Cit- (SEQ ID NO:11);

[1743] -Gly-Pro-Ser-Ala-Leu-Val-Cit- (SEQ ID NO:12);

[1744] -His-Val-Leu-Asn-Leu-Gly-Gly-Phe-Gly- (SEQ ID NO:13);

[1745] -Val-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:14);

[1746] -Ile-Pro-Val-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:15);

[1747] -Gly-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:16);

[1748] -Gly-Pro-Ala-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:17);

[1749] -Gly-Pro-Ala-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:18);

[1750] -Gly-Pro-Gln-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:19);

[1751] -Gly-Pro-Leu-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:20);

[1752] -Gly-Pro-Ser-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:21);

[1753] -Gly-Pro-Arg-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:22);

[1754] -Gly-Pro-Ala-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:23);

[1755] -Gly-Pro-Ser-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:24);

[1756] -Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:25);

[1757] -Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:26);

[1758] -Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:27);

[1759] -Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:28);

[1760] -Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:29);

[1761] -Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:30);

[1762] -Gly-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:31);

[1763] -Gly-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:32);

[1764] -Gly-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:33);

[1765] -Gly-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:34);

[1766] -Gly-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:35);

[1767] -His-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:36);

[1768] -His-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:37);

[1769] -His-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:38);

[1770] -His-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:39);

[1771] -His-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:40);

[1772] -His-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:41);

[1773] -Ser-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:42);

[1774] -Ser-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:43);

[1775] -Ser-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:44);

[1776] -Ser-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:45);

[1777] -Ser-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:46);

[1778] -Ser-Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:47);

[1779] -Ser-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:48);

[1780] -Gly-Pro-Ala-Pro-Leu-Val-Cit-(SEQ ID NO:49);

[1781] -Gly-Pro-Ala-Asn-Leu-Val-Cit-(SEQ ID NO:50);

[1782] -Gly-Pro-Arg-Gly-Dap(Me2)-Val-Cit-(SEQ ID NO:51);

[1783] or a variant having one or two mismatches relative to the sequence of any one of SEQ ID NOs: 1-51;

[1784] The D is camptothecin or its derivatives, auristatin or its derivatives, tetrahydroisoquinoline alkaloids or its derivatives, protein degraders and their derivatives, preferably DX8951, MMAE, Dxd, SN38, ET743, Lurbinectedin, CC885, CC-90009;

[1785] Said n represents an integer of 1-10.

[1786] In some embodiments, the antibody drug conjugate is selected from the following structures:

[1787] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-DX8951)n;

[1788] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLGLGGFG-DX8951)n;

[1789] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPASLVCit-PAB-DX8951)n;

[1790] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLVCit-PAB-DX8951)n;

[1791] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPSALVCit-PAB-DX8951)n;

[1792] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLGGFG-DX8951)n;

[1793] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAALGGFG-DX8951)n;

[1794] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-PRNLVCit-PAB-DX8951)n;

[1795] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRNLVCit-PAB-DX8951)n;

[1796] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGDap(Me2)LVCit-PAB-DX8951)n;

[1797] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-SPRNLVCit-PAB-DX8951)n;

[1798] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLSLVCit-PAB-DX8951)n;

[1799] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-MMAE)n;

[1800] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLVCit-PAB-MMAE)n;

[1801] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-PRNLVCit-PAB-MMAE)n;

[1802] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRNLVCit-PAB-MMAE)n;

[1803] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGDap(Me2)LVCit-PAB-MMAE)n;

[1804] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-SPRNLVCit-PAB-MMAE)n;

[1805] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLSLVCit-PAB-MMAE)n;

[1806] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-ET743)n;

[1807] Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRNLVCit-PAB-ET743)n;

[1808] The Ab represents a monoclonal antibody or an antigenic structural fragment thereof, or a modified monoclonal antibody or an antigen-binding fragment thereof, preferably anti-Her2, anti-TF, anti-ROR1, or anti-GPC3;

[1809] The DX8951 has the following structure:

[1810] in Indicates the connection location of DX8951;

[1811] The MMAE has the following structure:

[1812] in Indicates the connection position of MMAE;

[1813] The ET-743 has the following structure:

[1814] in Indicates the connection location of ET743;

[1815] Said n is an integer of 1-10, preferably, said n is 2, 4, 6, or 8.

[1816] In some embodiments, the antibody or antigen-binding fragment thereof is selected from the group consisting of: Trastuzumab, Tisotumab, Zilovertamab, Codrituzumab.

[1817] In some embodiments, the antibody drug conjugate is selected from:

[1818] Ab1-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-DX8951)n;

[1819] Ab1-((succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLGLGGFG-PAB-DX8951)n;

[1820] Ab1-((succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPASLVCit-PAB-DX8951)n;

[1821] Ab1-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLVCit-PAB-DX8951)n;

[1822] Ab1-((succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPSALVCit-PAB-DX8951)n;

[1823] Ab1-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLGGFG-PAB-DX8951)n;

[1824] Ab1-((succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAALGGFG-PAB-DX8951)n;

[1825] Ab1-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-PRNLVCit-PAB-DX8951)n;

[1826] Ab1-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRNLVCit-PAB-DX8951)n;

[1827] Ab1-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGDap(Me2)LVCit-PAB-DX8951)n;

[1828] Ab1-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-SPRNLVCit-PAB-DX8951)n;

[1829] The antibody Ab1 is trastuzumab; trastuzumab is a monoclonal antibody against Her2.

[1830] Said n is an integer of 1 to 10. Preferably, said n is 2, 4, 6 or 8.

[1831] In some embodiments, the antibody drug conjugate is selected from:

[1832] Ab2-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-DX8951)n;

[1833] Ab2-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLGLGGFG-PAB-DX8951)n;

[1834] Ab2-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLVCit-PAB-DX8951)n;

[1835] Ab2-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-PRNLVCit-PAB-DX8951)n;

[1836] Ab2-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRNLVCit-PAB-DX8951)n;

[1837] Ab2-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGDap(Me2)LVCit-PAB-DX8951)n;

[1838] Ab2-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-SPRNLVCit-PAB-DX8951)n;

[1839] Ab2-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-MMAE)n;

[1840] Ab2-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLVCit-PAB-MMAE)n;

[1841] Ab2-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-PRNLVCit-PAB-MMAE)n;

[1842] Ab2-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRNLVCit-PAB-MMAE)n;

[1843] Ab2-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGDap(Me2)LVCit-PAB-MMAE)n;

[1844] Ab2-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-SPRNLVCit-PAB-MMAE)n;

[1845] Ab2 is an anti-TF antibody, which may be a TisotumAb or a humanized anti-TF antibody, and n is an integer from 1 to 10. Preferably, n is 2, 4, 6, or 8.

[1846] In some embodiments, the antibody drug conjugate is selected from:

[1847] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-DX8951)n;

[1848] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLGLGGFG-PAB-DX8951)n;

[1849] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPASLVCit-PAB-DX8951)n;

[1850] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLVCit-PAB-DX8951)n;

[1851] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPSALVCit-PAB-DX8951)n;

[1852] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLGGFG-PAB-DX8951)n;

[1853] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAALGGFG-PAB-DX8951)n;

[1854] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-PRNLVCit-PAB-DX8951)n;

[1855] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRNLVCit-PAB-DX8951)n;

[1856] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGDap(Me2)LVCit-PAB-DX8951)n;

[1857] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-SPRNLVCit-PAB-DX8951)n;

[1858] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLSLVCit-PAB-DX8951)n;

[1859] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-MMAE)n;

[1860] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLVCit-PAB-MMAE)n;

[1861] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-PRNLVCit-PAB-MMAE)n;

[1862] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRNLVCit-PAB-MMAE)n;

[1863] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGDap(Me2)LVCit-PAB-MMAE)n;

[1864] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-SPRNLVCit-PAB-MMAE)n;

[1865] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLSLVCit-PAB-MMAE)n;

[1866] Ab3-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-ET743)n;

[1867] Wherein Ab3 is Codrituzumab, which is an anti-GPC3 antibody. Preferably, n is an integer from 1 to 10. Preferably, n is 2, 4, 6, 8

[1868] In some embodiments, the antibody drug conjugate is:

[1869] Ab4-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-DX8951)n;

[1870] Among them, Ab4 is Zilovertamab, which is an anti-ROR1 antibody.

[1871] Said n is an integer of 1 to 10. Preferably, said n is 2, 4, 6 or 8.

[1872] biological ligands

[1873] Antibodies or antigen-binding fragments thereof of the bioligands can be prepared using various methods known in the art, such as through genetic engineering and recombinant techniques. For example, DNA molecules encoding the heavy and light chain genes of the disclosed antibodies can be obtained by chemical synthesis or PCR amplification. The resulting DNA molecules are inserted into expression vectors, which are then transfected into host cells. The transfected host cells are then cultured under specific conditions to express the disclosed antibodies.

[1874] In some embodiments, the biological ligand is an anti-Her2 monoclonal antibody, such as trastuzumab and pertuzumab.

[1875] In some embodiments of the present application, the biological ligand is Trasuzumab, which is an anti-Her2 monoclonal antibody, and its amino acid sequence is known to those skilled in the art. Its schematic sequence can be found in, for example, patent CN103319599.

[1876] In some embodiments of the present application, the biological ligand is an anti-GPC3 antibody described in US Pat. No. 10,782,300B2 (i.e., Codrituzumab), which can be obtained by screening the disclosed vector design, construction, and antibody library displaying the antibody.

[1877] The heavy chain sequence and light chain amino acid sequence of codrituzumab monoclonal antibody can be found in SEQ ID No: 50 and SEQ ID No: 66 in US10782300B2, respectively.

[1878] In some embodiments of the present application, the biological ligand is an anti-ROR1 antibody, see SEQ ID No: 5 and SEQ ID No: 6 in US10335496B2.

[1879] In some embodiments of the present application, the biological ligand is an anti-TF antibody or an antigen-binding fragment thereof. In some embodiments, the anti-TF antibody includes all prior art anti-TF antibodies, such as Tisotumab, as well as anti-TF antibodies described in patents CN111818943A, CN106938051B, CN110575547A, CN103119065A, WO2011157741, WO 2010066803, and US 9168314 B2. In some embodiments, the anti-TF antibody or antigen-binding fragment thereof is selected from the group consisting of the anti-coagulant murine chimeric antibody 1849 described in US9920133B2 or the non-anti-coagulant murine chimeric antibody 1859 described in US9920133B2. In some embodiments, the anti-TF antibody is the TF antibody described in the fifth aspect below.

[1880] In a fifth aspect, the present application discloses an anti-tissue factor (TF) antibody or an antigen-binding fragment thereof, comprising:

[1881] (a) VH comprising the amino acid sequence of SEQ ID NO: 52 or 54, or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 52 or 54, or an amino acid sequence obtained by substitution, deletion, or addition of one or more amino acids to the amino acid sequence of SEQ ID NO: 52 or 54, which functions the same or similarly to the amino acid sequence of SEQ ID NO: 52 or 54; and / or

[1882] (b) VL comprising the amino acid sequence of SEQ ID NO: 53 or 55, or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 53 or 55, or an amino acid sequence obtained by substitution, deletion, or addition of one or more amino acids to the amino acid sequence of SEQ ID NO: 53 or 55, which functions the same or similarly to the amino acid sequence of SEQ ID NO: 53 or 55;

[1883] In some embodiments, the anti-TF antibody or antigen-binding fragment thereof is characterized in that the anti-TF antibody or antigen-binding fragment thereof comprises:

[1884] (1) VH and VL, wherein the VH has the amino acid sequence shown in SEQ ID NO: 52, and the VL has the amino acid sequence shown in SEQ ID NO: 53; or

[1885] (2) VH and VL, wherein the VH has the amino acid sequence shown in SEQ ID NO: 54, and the VL has the amino acid sequence shown in SEQ ID NO: 53; or

[1886] (3) VH and VL, wherein the VH has the amino acid sequence shown in SEQ ID NO: 54, and the VL has the amino acid sequence shown in SEQ ID NO: 55.

[1887] (4) VH and VL, wherein the VH has the amino acid sequence shown in SEQ ID NO: 52, and the VL has the amino acid sequence shown in SEQ ID NO: 55.

[1888] In some embodiments, the anti-TF antibody or antigen-binding fragment thereof comprises a light chain as set forth in SEQ ID NO: 56, or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 56; and a heavy chain as set forth in SEQ ID NO: 57, or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 57.

[1889] In some embodiments, the anti-TF antibody or antigen-binding fragment thereof comprises a light chain as set forth in SEQ ID NO: 56, or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 56; and a heavy chain as set forth in SEQ ID NO: 59, or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 59.

[1890] In some embodiments, the anti-TF antibody or antigen-binding fragment thereof comprises a light chain as set forth in SEQ ID NO: 58, or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 58; and a heavy chain as set forth in SEQ ID NO: 59, or an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 59.

[1891] In some embodiments, the TF is mammalian TF, and the antibody is preferably a mouse antibody, a monkey antibody, a rabbit antibody, a chimeric antibody, a humanized antibody, or a human antibody; and / or

[1892] The antibody is an IgG antibody, preferably an IgG1, IgG2, IgG3 or IgG4 antibody, more preferably an IgG1 antibody; and / or

[1893] The anti-TF antibody or antigen-binding fragment thereof is selected from the group consisting of a monoclonal antibody, a polyclonal antibody, a Fab fragment, a Fab' fragment, a F(ab')2 fragment, a Fd fragment, a Fv fragment, a dAb fragment, an isolated CDR region, a scFv and a nanobody.

[1894] In some embodiments, the TF is mammalian TF, and the antibody is preferably a modified mouse antibody, monkey antibody, rabbit antibody, chimeric antibody, humanized antibody, or human antibody; and / or

[1895] In some embodiments, the antibody is a modified IgG antibody, preferably a modified IgG1, IgG2, IgG3 or IgG4 antibody, more preferably a modified IgG1 antibody; and / or

[1896] In some embodiments, the anti-TF antibody or antigen-binding fragment thereof is selected from a modified monoclonal antibody, a polyclonal antibody, a Fab fragment, a Fab' fragment, a F(ab')2 fragment, a Fd fragment, a Fv fragment, a dAb fragment, an isolated CDR region, a scFv, and a nanobody.

[1897] In some embodiments, the modification methods include: amino acid point mutation, glycosylation modification, and polypeptide modification.

[1898] The antigen-binding fragments of the present application can be obtained by hydrolyzing intact antibody molecules (see Morimoio et al., J. Biochem. Biophys. Methods 24:107-117 (1992) and Brennan et al, Science 229:81 (1985)). In addition, these antigen-binding fragments can also be directly produced by recombinant host cells (see Verma et al. J. Immunol. Methods. 216:165-181 (1998); Little et al. Immunol. Today, 21:364-370 (2000)). For example, Fab' fragments can be obtained directly from host cells; Fab fragments can be chemically coupled to form F(ab')2 fragments (Carter et al, Biotechnology, 10:163-167 (1992)). In addition, Fv, Fab or F(ab')2 fragments can also be directly isolated from the culture medium of recombinant host cells. Other techniques for preparing such antigen-binding fragments are well known to those of ordinary skill in the art.

[1899] In a sixth aspect, the present disclosure provides a nucleic acid molecule encoding an anti-TF antibody or antigen-binding fragment thereof, or a heavy chain variable region and / or a light chain variable region thereof. Based on codon degeneracy known in the art, in certain embodiments, the nucleotide sequence can be substituted based on codon degeneracy.

[1900] In certain embodiments, the nucleotide sequence is codon-optimized.

[1901] In certain embodiments, the isolated nucleic acid molecules described in the present disclosure comprise: (i) a first nucleic acid and a second nucleic acid encoding the heavy chain variable region and the light chain variable region of the antibody or antigen-binding fragment thereof of the present disclosure, respectively, or (ii) a first nucleic acid encoding the heavy chain variable region and the heavy chain constant region, and a second nucleic acid encoding the light chain variable region and the light chain constant region, respectively, of the antibody or antigen-binding fragment thereof of the present disclosure, or (iii) a first nucleic acid and a second nucleic acid encoding the heavy chain and the light chain of the antibody or antigen-binding fragment thereof of the present disclosure, respectively.

[1902] In certain embodiments, the first and second nucleic acids comprise degenerate sequences or substantially identical sequences to any of the first and second nucleic acids in (i) to (iii) above. In certain embodiments, the degenerate sequences or substantially identical sequences refer to sequences having at least about 85%, 90%, 95%, 99% or higher sequence identity, or sequences having one or more nucleotide substitutions, or sequences that differ by no more than 3, 6, 15, 30, or 45 nucleotides compared to the nucleic acid molecules in (i) to (iii).

[1903] In a seventh aspect, a vector (eg, a cloning vector or an expression vector) is provided, comprising the nucleic acid molecule of the present application.

[1904] In certain embodiments, the vectors of the present disclosure are, for example, plasmids, cosmids, phages, lentiviruses, and the like.

[1905] In certain embodiments, the vector is capable of expressing the disclosed antibodies or antigen-binding fragments thereof in a subject (eg, a mammal, such as a human).

[1906] In an eighth aspect, the present application provides a recombinant cell comprising the nucleic acid molecule described above in the claims or a recombinant vector comprising the nucleic acid molecule.

[1907] Host cells can be eukaryotic cells (e.g., mammalian cells, insect cells, yeast cells) or prokaryotic cells (e.g., E. coli). Suitable eukaryotic cells include, but are not limited to, NSO cells, Vero cells, Hda cells, COS cells, CHO cells, HEK293 cells, BHK cells, and MDCKII cells. Suitable insect cells include, but are not limited to In certain embodiments, the host cell of the present application is a mammalian cell, such as a CHO cell (eg, CHOK1, CHO-S, CHODXB11, CHODG44).

[1908] In a ninth aspect, the present application provides a method for preparing an anti-TF antibody or an antigen-binding fragment thereof, the method comprising culturing the recombinant cell.

[1909] In a tenth aspect, the present application provides use of an anti-TF antibody or antigen-binding fragment thereof, nucleic acid molecule, recombinant vector or recombinant cell in the preparation of a medicament; preferably, the medicament is a medicament for preventing and / or treating cancer; more preferably, the cancer is a TF-expressing cancer.

[1910] In some embodiments, the anti-TF antibody or antigen-binding fragment thereof, nucleic acid molecule, recombinant vector or recombinant cell described above is used in the preparation of a reagent for diagnosing a cell proliferation-related disease; preferably, the disease is cancer; more preferably, the cancer is a TF-expressing cancer.

[1911] Preparation process

[1912] In an eleventh aspect, the present application provides a method for preparing a polypeptide or polypeptide fragment -P1-P2- or its tautomers, racemates, enantiomers, diastereomers, or mixed forms thereof, or an acceptable salt thereof, or a precursor thereof.

[1913] In a twelfth aspect, the present application provides a method for preparing the linker or its tautomer, mesomer, enantiomer, diastereomer, or mixed form thereof, or an acceptable salt thereof, or a precursor thereof.

[1914] In some embodiments, the method comprises sequentially linking the polypeptide or polypeptide fragment -P1-P2- to its preceding and following units C1, La, L1, and L2, wherein C1, La, L1, and L2 are as defined herein.

[1915] In a thirteenth aspect, the present application provides a method for preparing the linker drug or its tautomer, mesomer, enantiomer, diastereomer, or mixed form thereof, or an acceptable salt thereof, or a precursor thereof.

[1916] In some embodiments, the method comprises sequentially linking the polypeptide or polypeptide fragment -P1-P2- and its preceding and following units C1', La, L1, L2, and D, wherein C1', La, L1, L2, and D are as defined herein.

[1917] In a fourteenth aspect, the present application provides a method for preparing the conjugate as described above or its tautomer, mesomer, enantiomer, diastereomer, or mixed form thereof, or an acceptable salt thereof, or a precursor thereof.

[1918] In some embodiments, the method comprises sequentially linking Bp, C1 or C1', La, L1, L2, P1, P2, D, or precursors thereof.

[1919] Wherein said P1 and P2 are as defined above;

[1920] C1, La, L1, L2 are as defined above;

[1921] C1', D are as defined above;

[1922] Bp is as defined above.

[1923] In a fifteenth aspect, the present application provides a method for preparing the conjugate or its tautomer, mesomer, enantiomer, diastereomer, or mixed form thereof, or an acceptable salt thereof, or a precursor thereof.

[1924] In some embodiments, the method comprises coupling Bp with a linker drug C1'-La-L1-P1-P2-L2-D in a suitable solvent and conditions, wherein C1'-La-L1-P1-P2-L2-D is as defined above.

[1925] Preferably, the molar ratio of Bp to the linker drug C1'-La-L1-P1-P2-L2-D is 1:(1-20).

[1926] In some embodiments, the molar ratio of Bp to the linker drug C1'-La-L1-P1-P2-L2-D can be 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, or 1:12.

[1927] In some embodiments, preferably, the coupling reaction is carried out in water or a buffer salt system and / or an organic solvent;

[1928] In some embodiments, preferably, the buffer salt system is selected from PBS, sodium acetate buffer, histidine-hydrochloric acid buffer, histidine-acetate buffer, citric acid buffer, and the above buffers with added sucrose; more preferably, the buffer is selected from PBS pH 7.4, PBS pH 7.2, PBS pH 6.5, PBS pH 6.0, histidine-acetate pH 6.5, histidine-acetate pH 6.0, histidine-acetate pH 5.5 or a combination with 5-9% sucrose.

[1929] In some embodiments, the buffer system selected is histidine-acetate pH 5.5.

[1930] In some embodiments, the selected buffer system is PBS pH 6.0.

[1931] In some embodiments, the organic solvent is selected from N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, N-methylpyrrolidone, acetonitrile, methanol, ethanol, ethylene glycol, propylene glycol, and glycerol.

[1932] Preferably, the method further comprises the step of purifying the coupled product. Preferably, the purification is performed using one or more of an ultrafiltration system, gel chromatography, affinity chromatography, hydrophobic chromatography, reverse phase chromatography, or ion exchange chromatography. More preferably, the purification is performed using one or more of an ultrafiltration tube, a tangential flow system, a desalting column, a gel column, a cation exchange column, an anion exchange column, or an affinity column.

[1933] In the sixteenth aspect, the present application provides a pharmaceutical composition comprising the above-mentioned polypeptide or polypeptide fragment, linker, linker drug, conjugate, antibody or its antigen-binding fragment, nucleic acid molecule, recombinant vector, recombinant cell, and its tautomers, mesomers, racemates, enantiomers, diastereomers or their available salts, and optionally one or more pharmaceutical excipients.

[1934] In some embodiments, preferably, the composition further comprises a pharmaceutically acceptable carrier and / or excipient, for example, Tween 80, poloxamer 188, sucrose, trehalose, etc.

[1935] In the seventeenth aspect, the present application provides the use of the above-mentioned polypeptides or polypeptide fragments, linkers, linker drugs, conjugates, antibodies or antigen-binding fragments thereof, nucleic acid molecules, recombinant vectors, recombinant cells, and their tautomers, mesomers, racemates, enantiomers, diastereomers or their available salts in the preparation of drugs, which are used to treat / prevent diseases or conditions.

[1936] In the eighteenth aspect, the present application provides the use of any polypeptide or polypeptide fragment, any linker, any linker drug, conjugate, antibody or antigen-binding fragment thereof, nucleic acid molecule, recombinant vector, recombinant cell, and its tautomers, mesomers, racemates, enantiomers, diastereomers or their available salts in the preparation of a drug, wherein the drug is used to treat / prevent diseases related to abnormal cell activity.

[1937] In the nineteenth aspect, the present application provides the use of any one of the polypeptides or polypeptide fragments, linkers, linker drugs, conjugates, antibodies or antigen-binding fragments thereof, nucleic acid molecules, recombinant vectors, recombinant cells, and their tautomers, racemates, racemates, enantiomers, diastereomers or their available salts in the preparation of drugs, which are used to treat / prevent diseases related to abnormal cell activity.

[1938] In some embodiments, the medicament is for treating / preventing a disease or disorder.

[1939] In some embodiments, the medicament is used to treat / prevent diseases associated with abnormal cell activity.

[1940] In some embodiments, the disease or condition comprises a tumor.

[1941] In some embodiments, the disease or condition includes solid tumors and hematological tumors.

[1942] In a twentieth aspect, the present application provides a method for treating a disease or disorder in a subject in need thereof, comprising administering an effective amount of a conjugate to the subject.

[1943] In some embodiments, the tumor is selected from the following protein expression-related tumors: EGFR, HER2, HER3, c-Met, claudin18.2, TROP-2, folate receptor alpha (FRα), ROR1, ROR2, BCMA, PSMA, CD19, CD20, CD22, CD30, CD33, CD37, CD46, CD48, CD56, CD79b, CD123, CD138, CD166, CD276, CEACAM5, SDC1, CD74, CD70, MUC1, MUC16, GUCY2C, MSLN, SCL34A2, FOLH1, TPBC, PVRL4, STEAP1, SCL44A4, NACM1, EDNRB, GPNMB, FGFR, SEZ6, Nectin-4, PD-L1, Tissue Factor(TF), B7-H3, B7-H4, LY6E, LIV-1, CDH4, CDH6, ITGB6, GPC1, GPC3, ENPP3, KAAG1, FZD7, SSTR2, DLK1, TMEFF1, TM4SF1, SLAMF7, DLL3, FLT3, TNFRSF10B, EPCAM, AXL, IL2RA, 5T4, FAP, ICAM-1, IGF-1R;

[1944] Preferably, selected from tumors associated with Her2 expression;

[1945] Preferably, selected from tumors associated with Trop2 expression;

[1946] Preferably, it is selected from tumors associated with EGFR expression;

[1947] Preferably, it is selected from tumors related to FRα expression;

[1948] Preferably, it is selected from tumors associated with TF expression;

[1949] Preferably, it is selected from tumors associated with ROR1 expression;

[1950] Preferably, the tumor is selected from tumors associated with GPC3 expression.

[1951] In some embodiments, the tumor or cancer is selected from: esophageal cancer, brain cancer, lung cancer, epithelial cell cancer, bladder cancer, gastric cancer, ovarian cancer, pancreatic cancer, head and neck cancer, urothelial cancer, colon cancer, colorectal cancer, cervical cancer, endometrial cancer, rectal cancer, kidney cancer, prostate cancer, melanoma, liver cancer, gallbladder cancer, bile duct cancer, thyroid cancer, non-Hodgkin's lymphoma, multiple myeloma, B cell lymphoma.

[1952] Preferably, the tumor or cancer is selected from esophageal cancer, lung cancer, breast cancer, gastric cancer, urothelial cancer, bile duct cancer, prostate cancer, colorectal cancer, cervical cancer, and ovarian cancer.

[1953] In some embodiments, the conjugate can be administered by any conventional route known in the art, such as parenteral, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal administration.

[1954] Advantageous Effects of the Invention

[1955] This application designs a series of polypeptides that respond to both MMP-2 and / or MMP-9 specific to tumor tissues and Cathepsin B in tumor cells, as well as spacer structures connecting the polypeptides to antibodies and payloads, which can achieve at least one of the following technical effects.

[1956] 1. The peptide linker can be hydrolyzed by MMP-2 / MMP-9 and Cathepsin B simultaneously;

[1957] 2. The ADC prepared by this peptide linker can be cleaved by MMP-2 / MMP-9 or Cathepsin B enzymes to release the drug;

[1958] 3. ADCs prepared with peptide linkers are preferred to have better hydrophilicity than similar ADCs;

[1959] 4. ADCs prepared with preferred peptide linkers have better culture medium and plasma stability than similar ADCs;

[1960] 5. ADCs prepared with peptide linkers have better cell-killing activity than similar ADCs;

[1961] 6. ADCs prepared with preferred peptide linkers have better tumor suppression effects in CDX models compared to similar ADCs. BRIEF DESCRIPTION OF THE DRAWINGS

[1962] FIG1 shows the results of the endocytosis test of the anti-TF antibody of the present application on A431 cells after 4 hours.

[1963] FIG2 shows the growth inhibitory effect of h1849-H2L1-ADC-01 of the present application on the human prostate cancer DU145 tumor transplantation model.

[1964] FIG3 shows the growth inhibitory effect of h1849-H3L3-ADC-01 of the present application on the human prostate cancer DU145 tumor transplantation model.

[1965] FIG4 shows the growth inhibitory effect of h1849-H3L3-ADC-04 of the present application on the human prostate cancer DU145 tumor transplantation model.

[1966] FIG5 shows the growth inhibitory effect of h1849-H3L3-ADC-05 of the present application on the human cervical cancer Ca Ski tumor transplantation model.

[1967] FIG6 shows the growth inhibitory effect of the GC33-ADC of the present application on a human liver cancer HepG2 tumor transplantation model. DETAILED DESCRIPTION

[1968] The present invention will be further described below by way of specific embodiments, but this is not intended to limit the present disclosure. Those skilled in the art may make various modifications or improvements based on the teachings of the present invention without departing from the basic idea and scope of the present invention. Reagents or instruments used where the manufacturer is not indicated are conventional products that can be purchased commercially.

[1969] Sequence and its specific information

[1970] Example 1 Synthesis of polypeptide fragments, linkers, and linker drugs

[1971] Preparation plan

[1972] The structures of the compounds described in the following examples were determined by nuclear magnetic resonance ( 1HNMR) or mass spectrometry (MS). 1 H NMR was measured using a Bruker AVANCE NEO 400 MHz nuclear magnetic resonance instrument; the measurement solvents were deuterated methanol (CD, OD), deuterated chloroform (CDCl3), or hexadeuterated dimethyl sulfoxide (DMSO-d6); and the internal standard was tetramethylsilane (TMS).

[1973] The abbreviations used in the nuclear magnetic resonance (NMR) spectra used in the examples are shown below. s: singlet, d: doublet, t: triplet, q: quartet, dd: double doublet, qd: quartet doublet, ddd: double double doublet, ddt: double double triplet, dddd: double double double doublet, m: multiplet, br: broad, J: coupling constant, Hz: hertz, DMSO-d6: deuterated dimethyl sulfoxide. δ values ​​are expressed in ppm.

[1974] The mass spectrometry (MS) was performed using a SHIMADZU LCMS-2020 mass spectrometer.

[1975] 1. Synthesis of peptide fragments and bioactive molecules

[1976] 1.1 Synthesis of peptide fragments

[1977] Peptide list:

[1978] 1.1.1: Synthesis of 2-P1

[1979] The peptide was synthesized using standard Fmoc chemistry.

[1980] 1) Resin preparation: DMF was added to a container containing Rink Amide MBHA resin (concentration: 0.3 mmol / g, 0.1 mmol, 0.30 g) and allowed to swell for 2 hours.

[1981] 2) Deprotection: add 20% piperidine in DMF (10.0 mL), stir at 25° C. with N 2 for 15 min, wash with DMF (10.0 mL*5), and filter to obtain a resin.

[1982] 3) Coupling: HBTU (2.85 eq) and fmoc-citoh (3.00 eq) in DMF (10.0 mL) were added to the resin, followed by DIEA (6.00 eq). The mixture was stirred at 25°C with N2 for 30 min, and the resin was washed with DMF (10.0 mL*5).

[1983] 4) Repeat steps 2 to 3 above, using the following materials and reagents #1-#6 to couple amino acids in sequence

[1984] 5) Add 5% Ac2O, 10% NMM and DMF, and stir at room temperature for 30 min.

[1985] Peptide cleavage and purification:

[1986] 1) Add lysate (10.0 mL, 90.0% TFA / 2.50% TIS / 2.0% H2O / 5.0% DTT) to the resin flask and stir at room temperature for 2.5 hours.

[1987] 2) Precipitate the peptide with cold isopropyl ether (50 mL). Filter and collect the filter cake. Wash the filter cake with isopropyl ether (50 mL x 2). Vacuum dry the crude peptide for 2 hours to obtain the crude peptide (0.20 g).

[1988] 3) The crude peptide was purified by HPLC (purification conditions see the table below) to obtain the final product 2-P1(PRGLVCit) (7.4 mg, 9.97 μmol, 9.97% yield, 99.5% purity, TFA) as a white solid.

[1989] ESI-MS (m / z): 739.4 [M+H];

[1990] Purification conditions:

[1991] Other peptides were synthesized using the corresponding materials and reagents according to the above method, see the table below:

[1992] 1.2. Synthesis of Linkers

[1993] 1.2.1 Synthesis of L005

[1994] 1.2.1.1: Synthesis of (S)-2-((S)-2-(S)-2-[(S)-1-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycylamino)pyrrolidine-2-carboxamido)propionamido)acetamido)-4-methylpentanamido)-3-methylbutanamido)-5-ureidopentanic acid (1.2.1.1)

[1995] The peptide was synthesized using standard Fmoc chemistry.

[1996] 1) Under nitrogen bubbling, dichloromethane was added to a vessel containing CTC resin (2.0 mmol, 0.83 mmol / g, 2.4 g) and Fmoc-Cit-OH (0.8 g, 2.0 mmol / g, 1.0 eq).

[1997] 2) Diisopropylethylamine (6.0 equiv.) was added dropwise and mixed for 2 hours.

[1998] 3) Add methanol (2.4 mL) and mix for 0.5 hours.

[1999] 4) Drain the water and wash with DMF 5 times.

[2000] 5) Add 20% piperidine / DMF and react for 0.5 hours.

[2001] 6) Drain the water and wash with DMF 5 times.

[2002] 7) Add the Fmoc-amino acid solution and mix for 30 seconds, then add the corresponding coupling reagent. The reaction is continued for 1 hour under nitrogen bubbling.

[2003] 8) Repeat steps 4 to 7 for the next amino acid coupling.

[2004] 20% piperidine in DMF was used for Fmoc deprotection for 30 min. The coupling reaction was monitored by ninhydrin test and the resin was washed 5 times with N,N-dimethylformamide.

[2005] Peptide cleavage and purification:

[2006] 1) Add cleavage buffer (20% HFIP / DCM) to the flask containing the side-chain protected peptide at room temperature and stir for 0.5 hours*3.

[2007] 2) Filter and collect the filtrate.

[2008] 3) The solvent was removed under vacuum.

[2009] 4) The crude peptide was purified by preparative HPLC (A: 0.075% TFA in H2O, B: ACN) to afford compound 1.2.1.1 as a white solid (125 mg, 99.2%).

[2010] ESI-MS (m / z): 863.4 [M+H];

[2011] 1.2.1.2: Synthesis of (S)-1-((6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycylamino)-N-((S)-1-((2-(((S)-1-(S)-1-[(S)-1]((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amine)-4-methyl-1-oxopentan-2-yl)amino)-2-oxoethyl)amino)-1-oxopropan-2-yl)pyrrolidine-2-carboxamide (L005)

[2012] To a solution of compound 1.2.1.1 (50.0 mg, 57.9 μmol) and (4-aminophenyl)methanol (7.14 mg, 57.9 μmol) in pyridine (1 mL) was added EDCI (44.4 mg, 231 μmol). The mixture was stirred at 20°C for 1 hour. LC-MS confirmed the reaction was complete. The reaction solution was added dropwise to isopropyl ether (10 ml * 1) to obtain a solid. The residue was purified by preparative HPLC to obtain compound L005 (16.0 mg, 14.8 μmol, 25.7% yield, 90.1% purity) as a pink solid.

[2013] ESI-MS (m / z): 968.3 [M+H] + ;

[2014] 1.2.2 Synthesis of L006

[2015] 1.2.2.1 Synthesis of (6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycyl-L-prolyl-L-leucylglycyl-L-leucylglycyl-L-phenylalanylglycine (L006)

[2016] The peptide was synthesized using standard Fmoc chemistry.

[2017] 1) Add dichloromethane to a container containing CTC resin (1.0 mmol, 0.83 mmol / g, 1.2 g) and Fmoc-Gly-OH (0.3 g, 1.0 mmol, 1.0 eq) and bubble nitrogen through the mixture.

[2018] 2) Diisopropylethylamine (6.0 eq) was gradually added and mixed for 2 hours.

[2019] 3) Methanol (1.2 mL) was added and stirred for 0.5 hours.

[2020] 4) Drain and wash with DMF 5 times.

[2021] 5) Add 20% piperidine / DMF and react for 0.5 hours.

[2022] 6) Drain and wash with DMF 5 times.

[2023] 7) Add the Fmoc-amino acid solution and mix for 30 seconds, then add the corresponding coupling reagent. The reaction continues for 1 hour with nitrogen bubbling.

[2024] 8) Repeat steps 4 to 7 to couple the next amino acid.

[2025] 20% piperidine in DMF was used for Fmoc deprotection for 30 min. The coupling reaction was monitored by ninhydrin test and the resin was washed 5 times with DMF.

[2026] Peptide cleavage and purification:

[2027] 1) Add lysis buffer (20% HFIP / DCM) to the flask containing the side chain protected peptide and stir at room temperature for 0.5 h*3.

[2028] 2) Filter and collect the filtrate.

[2029] 3) The solvent was removed under vacuum.

[2030] 4) The crude peptide was purified by HPLC (A: 0.075% TFA in H2O, B: ACN) to obtain compound L006 (134 mg, 99.5%) as a white solid.

[2031] ESI-MS (m / z): 967.5 [M+H] + .

[2032] 1.2.3 Synthesis of L011

[2033] 1.2.3.1 Synthesis of (6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycyl-L-prolyl-L-arginylglycyl-L-leucylglycylglycyl-L-phenylalanylglycine (L011)

[2034] The peptide was synthesized using standard Fmoc chemistry.

[2035] 1) Add dichloromethane to a container containing CTC resin (1.0 mmol, 0.83 mmol / g, 1.2 g) and Fmoc-Gly-OH (1.0 mmol, 1.0 eq) and bubble nitrogen through the container.

[2036] 2) Diisopropylethylamine (6.0 eq) was gradually added and mixed for 2 hours.

[2037] 3) Methanol (1.3 mL) was added and stirred for 0.5 h.

[2038] 4) Drain and wash with DMF 3 times.

[2039] 5) Add 20% piperidine / DMF and react for 30 minutes.

[2040] 6) Drain and wash with DMF 5 times.

[2041] 7) Add the Fmoc-amino acid solution and mix for 30 seconds, then add the corresponding coupling reagent. The reaction continues for 1 hour with nitrogen bubbling.

[2042] 8) Repeat steps 4 to 7 to couple the next amino acid.

[2043] 20% piperidine in DMF was used for Fmoc deprotection for 30 min. The coupling reaction was monitored by ninhydrin test and the resin was washed 5 times with DMF.

[2044] Peptide cleavage and purification:

[2045] 1) The peptide resin obtained above was washed three times with methanol and dried in vacuo.

[2046] 2) Add cleavage buffer (50% TFA / DCM) to the peptide resin and stir for 1 hour.

[2047] 3) DCM and TFA were concentrated under reduced pressure.

[2048] 4) Vacuum drying for 2 hours to obtain crude peptide (1.0 g).

[2049] 5) The crude peptide was purified by HPLC (A: 0.075% TFA in H2O, B: ACN) to obtain compound L011 (100 mg, purity 99.1%) as a white solid.

[2050] ESI-MS (m / z): 1010.4 [M+H] + ;

[2051] 1.2.4 Synthesis of L012

[2052] 1.2.4.1 Synthesis of (6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycyl-L-prolyl-L-alanyl-glycyl-L-leucylglycylglycyl-L-phenylalanylglycine (L012)

[2053] The peptide was synthesized using standard Fmoc chemistry.

[2054] 1) DCM was added to a vessel containing CTC resin (1.0 mmol, 0.83 mmol / g, 1.2 g) and Fmoc-Gly-OH (1.0 mmol, 1.0 eq) and N2 was bubbled through the vessel.

[2055] 2) Diisopropylethylamine (6.0 eq) was gradually added and mixed for 2 hours.

[2056] 3) Methanol (1.3 mL) was added and stirred for 0.5 h.

[2057] 4) Drain and wash with DMF 3 times.

[2058] 5) Add 20% piperidine / DMF and react for 30 minutes.

[2059] 6) Drain and wash with DMF 5 times.

[2060] 7) Add fmoc-amino acid solution and mix for 30 seconds, then add corresponding coupling reagent. The reaction continues for 1 hour with nitrogen bubbling.

[2061] 8) Repeat steps 4 to 7 to couple the next amino acid.

[2062] 20% piperidine in DMF was used for Fmoc deprotection for 30 min. The coupling reaction was monitored by ninhydrin test and the resin was washed 5 times with DMF.

[2063] Peptide cleavage and purification:

[2064] 1) The peptide resin obtained above was washed three times with methanol and dried in vacuo.

[2065] 2) Add lysis buffer (20% HFIP / DCM) to the peptide resin and stir for 30 min*3 times.

[2066] 3) DCM was concentrated under reduced pressure.

[2067] 4) Vacuum drying for 2 hours to obtain crude peptide (0.9 g).

[2068] 5) The crude peptide was purified by HPLC (A: 0.075% TFA in H2O, B: ACN) to obtain compound L012 (70 mg, 99.8%) as a white solid.

[2069] ESI-MS (m / z): 939.4 [M+H] + .

[2070] Other linkers can be synthesized using the corresponding raw materials according to the above method.

[2071] 1.3 Synthesis of linker fragments

[2072] 1.3.1 Synthesis of (S)-2-((S)-4-amino-2-((S)-2-(S)-1-((6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycyl)pyrrolidine-2-carboxamido)-5-guanidinopentanamido)-4-oxobutanamido)-3-(dimethylamino)propanoic acid (1.3.1)

[2073] The peptide was synthesized using standard Fmoc chemistry.

[2074] 1) Under nitrogen bubbling, DCM was added to a vessel containing CTC resin (1.0 mmol, 1.6 g, Sub = 0.64 mmol / g) and (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(dimethylamino)propanoic acid (0.35 g, 1.0 mmol, 1 eq.).

[2075] 2) Add DIEA (4.0 eq.) dropwise and mix for 2 hours.

[2076] 3) Add methanol (1.5 mL) and mix for 30 minutes.

[2077] 4) Drain the water and wash with DMF 5 times.

[2078] 5) Add 20% piperidine / DMF and react for 30 minutes.

[2079] 6) Drain the water and wash with DMF three times.

[2080] 7) Add Fmoc amino acid solution and mix for 30 seconds, then add the corresponding coupling reagent and bubble N2 for about 1 hour.

[2081] 8) Repeat steps 5 to 7 for the next amino acid coupling.

[2082] Fmoc deprotection was performed using 20% ​​piperidine in DMF for 30 minutes. The coupling reaction was monitored by ninhydrin test and the resin was washed 5 times with DMF.

[2083] Peptide cleavage and purification:

[2084] 1) The as-received resin was washed with MeOH three times and dried in vacuo.

[2085] 2) Add lysis buffer (20% HFIP / DCM) to the peptide resin and stir for 0.5 hours three times.

[2086] 3) DCM was concentrated under reduced pressure.

[2087] 4) The peptide was dried under vacuum for 2 hours to obtain compound 1.3.1 (55 mg, 87.79%).

[2088] ESI-MS (m / z): 750.3 [M+H] + ;

[2089] Other linker fragments were synthesized using the following raw materials according to the above method, as shown in the table below:

[2090] 1.4 Synthesis of Linker-Payload Intermediates

[2091] Linker drug list:

[2092] 1.4.1 Synthesis of L005-P005

[2093] 1.4.1.1: Synthesis of 4-((3S,9S,12S,15S)-1-((S)-1-((6-(2,5-dioxy-2,5-dihydro-1H-pyrrolidin-1-yl)hexenyl)pyrrolidin-2-yl)-9-isobutyl-12-isopropyl-3-methyl-1,4,7,10,13-pentaoxy-15-(3-ureidopropyl)-2,5,8,11,14-pentaazahexadec-16-amino)benzyl (4-nitrophenyl) carbonate (1.4.1.1)

[2094] To a solution of compound L005 (16.0 mg, 16.5 μmol) and bis(4-nitrobenzene) carbonate (15.1 mg, 49.6 μmol) in DMF (200 μL) was added DIEA (8.54 mg, 66.1 μmol, 11.5 μL). The mixture was stirred at 25°C for 4 hours. LC-MS confirmed the reaction was complete. The reaction solution was added dropwise to isopropyl ether (5 ml x 1) to yield a solid. The crude product, compound 1.4.1.1 (16.0 mg), was a red solid and was used directly in the next step without further purification.

[2095] ESI-MS (m / z): 1133.3 [M+H] + .

[2096] 1.4.1.2: 4-((3S,9S,12S,15S)-1-((S)-1-)((6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycol)pyrrolidin-2-yl)-9-isobutyl-12-isopropyl-3-methyl-1,4,7,10,13-pentaoxo-15-(3-ureidopropyl)-2,5,8,11, Synthesis of 14-pentaazahexadecane-16-amino)benzyl(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (L005-P005)

[2097] Compound 1.4.1.1 (16.0 mg) and exatecan mesylate (6.00 mg, 11.3 μmol) were dissolved in DMF (200 μL) and HOBt (2.29 mg, 16.9 μmol) and DIEA (7.30 mg, 56.5 μmol, 9.84 μL) were added, respectively. The mixture was stirred at 25°C for 2 hours. LC-MS confirmed the reaction was complete. High-performance liquid chromatography (HPLC) was used for preparative purification to obtain compound L005-P005 as a yellow solid (6.30 mg, 4.10 μmol, yield: 30.9%, purity: 99.0%). ESI-MS (m / z): 1430.3 [M+H] + .

[2098] 1.4.2 Synthesis of L005-P002

[2099] 1.4.2.1: Synthesis of (3-ureidopropyl)-2,5,8,11,14-pentaazahexadecane-16-amido)benzyl(((S)-1-(((3R,4S,5S)-1-(S)-2-(1R,2R)-3-((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl1-oxobutan-2-yl(methyl)carbamate (L005-P002)

[2100] HOBt (5.44 mg, 40.2 μmol) and DIEA (8.67 mg, 67.1 μmol) were added to a solution of compound 1.4.1.1 (38 mg, 33.5 μmol) and MMAE (24.1 mg, 33.4 μmol) in DMF (400 μL). The mixture was stirred at 25°C for 8 hours. LC-MS confirmed the reaction was complete. High-performance liquid chromatography (HPLC) was used to preparatively purify the product to afford L005-P002 (25 mg, 14.6 μmol, 43.5% yield) as a yellow solid.

[2101] ESI-MS (m / z): 1712.6 [M+H] + .

[2102] 1.4.3 Synthesis of L005-P003

[2103] 1.4.3.1: Synthesis of L005-P003

[2104] To a solution of compound 1.4.1.1 (111 mg, 98.4 μmol) and ET743 (15 mg, 19.7 μmol) in DMF (0.5 mL) was added DIEA (12.7 mg, 17.1 μl, 298.4 μmol), followed by degassing and purging with N2 three times, and then the mixture was stirred at 25 ° C for 0.5 hours under N2 atmosphere. LC-MS detection showed that the reaction was complete. The mixture was added to stirred cold water (10 mL) and stirred at 25 ° C for 5 minutes. The residue solution was purified by HPLC to give L005-P003 (26 mg, yield: 71.6%) as a white solid. ESI-MS (m / z): 1737.8 [M-H2O] + .

[2105] 1.4.4 Synthesis of L006-P005

[2106] 1.4.4.1: (S)-1-((6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycylamino)-N-((S)-1-(((4S,13S)-13-(1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13,15-hexahydro-1H,12H-benzo[de]pyranoyl)- Synthesis of [3',4':6,7]indolizino[1,2-b]quinolin-1-yl)amino)-2-oxoethyl)carbamoyl)-4-isobutyl-2,5,8,11-tetraoxo-14-phenyl-3,6,9,12-tetraazatetradecyl)amino)-4-methyl-1-oxopentan-2-yl)pyrrolidine-2-carboxamide (L006-P005)

[2107] Compound L006 (30.0 mg, 31.0 μmol) and exatecan mesylate (13.2 mg, 24.8 μmol) were dissolved in DMF (300 μL). HOBt (10.1 mg, 74.4 μmol), DIC (15.6 mg, 124 μmol, 19.2 μL), and DIEA (20.1 mg, 155 μmol, 27.0 μL) were added, respectively. The mixture was stirred at 25°C for 3 hours. LC-MS confirmed the reaction was complete. High-performance liquid chromatography (HPLC) was used for preparative purification to obtain L006-P005 (13.0 mg, 9.39 μmol, yield: 30.2%) as a yellow solid. ESI-MS (m / z): 1384.3 [M+H] + .

[2108] 1.4.5 Synthesis of L008-P005

[2109] 1.4.5.1 Synthesis of (6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycyl-L-prolyl-L-alanyl-L-seryl-L-leucine (1.4.5.1)

[2110] The peptide was synthesized using standard Fmoc chemistry.

[2111] 1) DCM was added to a vessel containing CTC resin (2.0 mmol, 0.83 mmol / g, 2.4 g) and Fmoc-Leu-OH (2.0 mmol, 1.0 eq) and N2 was bubbled through the vessel.

[2112] 2) DIEA (6.0 eq) was gradually added and mixed for 2 hours.

[2113] 3) Add MeOH (2.5 mL) and stir for 0.5 h.

[2114] 4) Drain and wash with DMF 3 times.

[2115] 5) Add 20% piperidine / DMF and react for 30 minutes.

[2116] 6) Drain and wash with DMF 5 times.

[2117] 7) Add the Fmoc-amino acid solution and mix for 30 seconds, then add the corresponding coupling reagent. The reaction continues for 1 hour with nitrogen bubbling.

[2118] 8) Repeat steps 4 to 7 to couple the next amino acid.

[2119] 20% piperidine in DMF was used for Fmoc deprotection for 30 min. The coupling reaction was monitored by ninhydrin test and the resin was washed 5 times with DMF.

[2120] Peptide cleavage and purification:

[2121] 1) The peptide resin obtained above was washed three times with methanol and dried in vacuo.

[2122] 2) Add cleavage buffer (50% TFA / DCM) to the peptide resin and stir for 1 hour.

[2123] 3) DCM and TFA were concentrated under reduced pressure.

[2124] 4) Vacuum drying for 2 hours to obtain crude peptide (1.2 g).

[2125] 5) The crude peptide was purified by pre-HPLC (A: 0.075% TFA in H2O, B: ACN) to obtain compound 1.4.5.1 (110 mg, 99.8%) as a white solid.

[2126] ESI-MS (m / z): 637.3 [M+H] + ;

[2127] 1.4.5.2 Synthesis of (9H-fluoren-9-yl)methyl ((S)-1-((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate (1.4.5.2)

[2128] To a solution of compound Fmoc-Val-Cit-OH (20.0 g, 40.3 mmol, 1.0 eq.) in DCM (200 mL) and MeOH (50.0 mL) was added EEDQ (19.9 g, 80.5 mmol, 2.0 eq.) and PAB (9.92 g, 80.5 mmol, 2.0 eq.). The mixture was stirred at 25 ° C for 16 hours. LC-MS detected the completion of the reaction. The reaction product was filtered, and the filtrate was added to 2000 mL of EtOAc / MTBE (8 / 1) and stirred for 10 min twice, filtered, and the filtrate was washed with EtOAc to give white solid compound 1.4.5.2 (19.8 g, crude product). ESI-MS (m / z): 602.3 [M+H] + .

[2129] 1.4.5.3 Synthesis of (9H-fluoren-9-yl)methyl((S)-3-methyl-1-(((S)-1-((4-(((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)1-oxobutan-2-yl)carbamate (1.4.5.3)

[2130] To a solution of compound 1.4.5.2 (20.0 g, 33.2 mmol, 1.0 eq.) and PNP (20.2 g, 66.4 mmol, 2.0 eq.) in DMF (200 mL) was added DIEA (8.59 g, 66.4 mmol, 11.5 mL, 2.0 eq.). The mixture was stirred at 25 ° C for 3 hours. LC-MS detected that the reaction was complete. The reaction mixture was added dropwise to stirred ice MTBE and filtered. The filtered solid was dissolved in DCM and then poured into MTBE / EtOAc (1: 1) and stirred for 30 min. Then it was filtered and concentrated under reduced pressure to give compound 1.4.5.3 (21 g, crude product) as a white solid. ESI-MS (m / z): 767.4 [M + H] + .

[2131] 1.4.5.4 Synthesis of (9H-fluoren-9-yl)methyl((S)-1-((S)-1-(4-(1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate (1.4.5.4)

[2132] Compound 1.4.5.3 (3.00 g, 3.91 mmol, 1.0 eq), Exatecan mesylate (1.87 g, 3.52 mmol, 0.8 eq) and HOBt (635 mg, 4.70 mmol, 1.2 eq) were dissolved in DMF (30 mL) and DIEA (1.01 g, 7.83 mmol, 1.29 mL, 2.0 eq) was added to the mixture. The mixture was stirred at 25 ° C under N2 atmosphere for 3.0 hours. LC-MS detected the completion of the reaction. The mixture was triturated with isopropyl ether (300 mL) to give compound 1.4.5.4 (5.60 g, crude product) as a brown solid. ESI-MS (m / z): 1063.4 [M+H] + .

[2133] 1.4.5.5 Synthesis of 4-(((S)-2-[(S)-2-amino-3-methylbutanamido)-5-ureidopentanamido)benzyl((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (1.4.5.5)

[2134] A mixture of compound 1.4.5.4 (5.60 g, crude) in DMF (40 mL) was added to TEA (10 mL). The mixture was stirred at 25°C for 16 hours. LC-MS confirmed the reaction was complete. The reaction mixture was purified by preparative HPLC to afford compound 1.4.5.5 (2.75 g, 3.27 mmol, yield: 83.5%) as a yellow solid. ESI-MS (m / z): 841.6 [M+H] + .

[2135] 1.4.5.6 4-((3S,6S,9S,12S,15S)-1-((S)-1-)((6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycol)pyrrolidin-2-yl)-6-(hydroxymethyl)-9-isobutyl-12-isopropyl-3-methyl-1,4,7,10,13-pentaoxo-15-(3-ureidopropyl)-2,5, Synthesis of 8,11,14-pentaazahexadecyl)benzyl(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (L008-P005)

[2136] Compound 1.4.5.1 (30.0 mg, 47.1 μmol), compound 1.4.5.5 (39.6 mg, 47.1 μmol), PYBOP (49.0 mg, 94.2 μmol), and DIEA (32.8 μL, 188 μmol) were added to DMF (0.5 mL). The mixture was stirred at 25°C for 1 hour. LC-MS confirmed the reaction was complete. The crude product was purified by HPLC to obtain compound L008-P005 (8.0 mg, yield: 11.6%) as a yellow solid. ESI-MS (m / z): 1459.4 [M+H] + .

[2137] 1.4.6 Synthesis of L009-P005

[2138] 1.4.6.1 Synthesis of (6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycyl-L-prolyl-L-arginylglycyl-L-leucine (1.4.6.1)

[2139] The peptide was synthesized using standard Fmoc chemistry.

[2140] 1) Add dichloromethane to a container containing CTC resin (2.0 mmol, 0.83 mmol / g, 2.4 g) and Fmoc-Leu-OH (2.0 mmol, 1.0 eq) and bubble nitrogen through the container.

[2141] 2) Diisopropylethylamine (6.0 eq) was gradually added and mixed for 2 hours.

[2142] 3) Methanol (2.5 mL) was added and stirred for 0.5 h.

[2143] 4) Drain and wash with DMF 3 times.

[2144] 5) Add 20% piperidine / DMF and react for 30 minutes.

[2145] 6) Drain and wash with DMF 5 times.

[2146] 7) Add fmoc-amino acid solution and mix for 30 seconds, then add corresponding coupling reagent. The reaction continues for 1 hour with nitrogen bubbling.

[2147] 8) Repeat steps 4 to 7 to couple the next amino acid.

[2148] 20% piperidine in DMF was used for Fmoc deprotection for 30 min. The coupling reaction was monitored by ninhydrin test and the resin was washed 5 times with DMF.

[2149] Peptide cleavage and purification:

[2150] 1) The polypeptide resin obtained above was washed with methanol three times and vacuum dried.

[2151] 2) Add lysis buffer (50% TFA / DCM) to the peptide resin and stir for 1 hour.

[2152] 3) DCM and TFA were concentrated under reduced pressure.

[2153] 4) Vacuum drying for 2 hours to obtain crude peptide (1.4 g).

[2154] 5) The crude peptide was purified by HPLC (A: 0.075% TFA in H2O, B: ACN) to obtain compound 1.4.6.1 (120 mg, 99.0%) as a white solid.

[2155] ESI-MS (m / z): 692.4 [M+H] + .

[2156] 1.4.6.2 4-((6S,12S,15S,18S)-1-amino-6-((S)-1-((6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycylamino)pyrrolidine-2-carboxamido)-1-imino-12-isobutyl-15-isopropyl-7,10,13,6-tetraoxo-18-(3-ureidopropyl)-2,8,11,14 Synthesis of ((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (L009-P005)

[2157] Compound 1.4.6.1 (30.0 mg, 43.4 μmol), compound 1.4.5.5 (36.5 mg, 43.4 μmol), PYBOP (45.1 mg, 86.7 μmol), and DIEA (30.2 μL, 173 μmol) were added to DMF (0.5 mL). The mixture was stirred at 25°C for 1 hour. LC-MS confirmed the reaction was complete. The crude product was purified by HPLC to obtain compound L009-P005 (7.0 mg, yield: 10.6%) as a yellow solid. ESI-MS (m / z): 1515.4 [M+H] + .

[2158] 1.4.7 Synthesis of L009-P002

[2159] 1.4.7.1 4-((6S,12S,15S,18S)-1-amino-6-((S)-1-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycyl)pyrrolidine-2-carboxamido)-1-imino-12-isobutyl-15-isopropyl-7,10,13,16-tetraoxo-18-(3-ureidopropyl)-2,8,11,14,17-pentaazanonadecane Synthesis of (L009-P002)

[2160] To a solution of compound 1.4.6.1 (30.0 mg, 43.4 mmol) and Val-Cit-PAB-MMAE (48.7 mg, 43.4 mol) in DMF (0.3 mL) were added PYBOP (45.1 mg, 86.7 mmol) and DIEA (30.2 μL, 173 mmol). The mixture was stirred at 25°C for 1.5 hours. LC-MS confirmed the reaction was complete. Purification by preparative HPLC afforded compound L009-P002 (24.0 mg, 30.8% yield) as a white solid.

[2161] ESI-MS (m / z): 1798.6 [M+H] + ; 1H NMR: (400MHz, DMSO-d6)δ:10.02(br s,1H)8.23-8.40(m,1H)8.04-8.14(m,2H)7.86-8.01(m,4H)7.78-7.85(m,1H)7.54-7.60(m,2H)7.38(br t,J=5.75Hz,1H)7.23-7.35(m,7H)7.13-7.22(m,2H)7.00(s,2H)5.98(br s,1H)5.38-5.48(m,1H)4.91-5.15(m,3H)4.67-4.79(m,1H)4.48(br d,J=5.75Hz,1H)4.43(br d,J=6.63Hz,1H)4.30-4.40(m,4H)4.20-4.29(m,2H)4.16(br t,J=7.75Hz,1H)3.95-4.03(m,2H)3.92(br d,J=5.63Hz,1H)3.78-3.87(m,2H)3.69-3.77(m,3H)3.29-3.37(m,7H)3.23(d,J=6.13Hz,4H)3.19(d,J=10.13Hz,3H)3.11(br s,2H)3.05-3.10(m,2H)2.97(br s,1H)2.86(br d,J=14.76Hz,3H)2.41(br d,J=15.63Hz,1H)2.26(br dd,J=16.01,9.26Hz,1H)2.08-2.15(m,4H)1.94-2.05(m,4H)1.69-1.78(m,4H)1.51-1.62(m,5H)1.43-1.51(m,10H)1.23(br s,4H)0.96-1.06(m,7H)0.74-0.89(m,33H).

[2162] 1.4.8 Synthesis of L010-P005

[2163] 1.4.8.1 Synthesis of (6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycyl-L-prolyl-L-seryl-L-alanyl-L-leucine (1.4.8.1)

[2164] The peptide was synthesized using standard Fmoc chemistry.

[2165] 1) Add dichloromethane to a container containing CTC resin (2.0 mmol, 0.83 mmol / g, 2.4 g) and Fmoc-Leu-OH (2.0 mmol, 1.0 eq) and bubble nitrogen through the container.

[2166] 2) Diisopropylethylamine (6.0 eq) was gradually added and mixed for 2 hours.

[2167] 3) Add methanol (2.5 mL) and stir for 0.5 h.

[2168] 4) Drain and wash with DMF 3 times.

[2169] 5) Add 20% piperidine / DMF and react for 30 minutes.

[2170] 6) Drain and wash with DMF 5 times.

[2171] 7) Add the Fmoc-amino acid solution and mix for 30 seconds, then add the corresponding coupling reagent. The reaction continues for 1 hour with nitrogen bubbling.

[2172] 8) Repeat steps 4 to 7 to couple the next amino acid.

[2173] 20% piperidine in DMF was used for Fmoc deprotection for 30 min. The coupling reaction was monitored by ninhydrin test and the resin was washed 5 times with DMF.

[2174] Peptide cleavage and purification:

[2175] 1) The peptide resin obtained above was washed three times with methanol and dried in vacuo.

[2176] 2) Add cleavage buffer (50% TFA / DCM) to the peptide resin and stir for 1 hour.

[2177] 3) DCM and TFA were concentrated under reduced pressure.

[2178] 4) Vacuum drying for 2 hours to obtain crude peptide (1.2 g).

[2179] 5) The crude peptide was purified by HPLC (A: 0.075% TFA in H2O, B: ACN) to obtain compound 1.4.8.1 (90 mg, 99.6%) as a white solid.

[2180] ESI-MS (m / z): 637.3 [M+H] + .

[2181] 1.4.8.2 4-((3S,6S,9S,12S,15S)-1-((S)-1-)((6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycol)pyrrolidin-2-yl)-3-(hydroxymethyl)-9-isobutyl-12-isopropyl-6-methyl-1,4,7,10,13-pentaoxo-15-(3-ureidopropyl)-2,5, Synthesis of 8,11,14-pentaazahexadecyl)benzyl(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (L010-P005)

[2182] Compound 1.4.8.1 (30.0 mg, 47.1 μmol) and compound 1.4.5.5 (39.6 mg, 47.1 μmol), PYBOP (49.0 mg, 94.2 μmol), and DIEA (32.8 μL, 188 μmol) were added to DMF (0.5 mL). The mixture was stirred at 25°C for 1 hour. LC-MS confirmed the reaction was complete. The crude product was purified by HPLC to obtain compound L010-P005 (9.0 mg, 13.1% yield) as a yellow solid.

[2183] ESI-MS (m / z): 1460.2 [M+H] + .

[2184] 1.4.9 Synthesis of L011-P005

[2185] 1.4.9.1(S)-1-((6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycylamino)-N-((S)-1-((4S,13S)-13-(2-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12 Synthesis of H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)amino)-2-oxoethyl)carbamoyl)-4-isobutyl-2,5,8,11-tetraoxo-14-phenyl-3,6,9,12-tetraazatetradecyl)amino)-5-guanidine-1-oxopentan-2-yl)pyrrolidine-2-carboxamide (L011-P005)

[2186] Compound L011 (30.0 mg, 29.7 μmol) and exatecan mesylate (12.6 mg, 23.8 μmol) were dissolved in DMF (300 μL), and HOBt (9.63 mg, 71.3 μmol), DIC (18.4 μL, 119 μmol), and DIEA (20.7 μL, 118.80 μmol) were added, respectively. The mixture was stirred at 25°C for 8 hours. LC-MS confirmed the reaction was complete. The reaction mixture was acidified with acetic acid and purified by HPLC. Compound L011-P005 (6.0 mg, 14.2% yield) was obtained as a yellow solid. ESI-MS (m / z): 1428.4 [M+H] + .

[2187] 1.4.10 Synthesis of L012-P005

[2188] 1.4.10.1(S)-1-((6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycylamino)-N-((S)-1-(((S)-1-((2-((2-(((S)-1-((2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H Synthesis of (4-(amino)-1-oxopropyl)-2-nitro-1-thiazolyl-1-ol)-1-nitro-1-thiazolyl-1-ol)-1-nitro-1-thiazolyl-1-ol)-1-nitro-1-thiazolyl-1-ol)-1-nitro-1-thiazolyl-1-ol)-1-nitro-1-thiazolyl-1-ol)-1-nitro-1-thiazolyl-1-ol)-1-nitro-1-thiazolyl-1-ol)-1-nitro-1-thiazolyl-1-ol)-1-nitro-1-thiazolyl-1-ol)-1-nitro-1-thiazolyl-1-ol)-1-nitro-1-thiazolyl-1-ol)-1-nitro-1-thiazolyl-1-ol)

[2189] Compound L012 (30.0 mg, 32.0 μmol) and exatecan mesylate (13.6 mg, 25.6 μmol) were dissolved in DMF (300 μL), and HOBt (10.4 mg, 76.7 μmol), DIC (19.8 μL, 128 μmol), and DIEA (22.3 μL, 128 μmol) were added, respectively. The mixture was stirred at 25°C for 8 hours. LC-MS confirmed the reaction was complete. The reaction mixture was acidified with acetic acid and purified by HPLC. Compound L012-P005 (8.0 mg, yield: 18.5%) was obtained as a yellow solid. ESI-MS (m / z): 1357.4 [M+H] + .

[2190] 1.4.11 Synthesis of L013-P005

[2191] 1.4.11.1 4-((6S,9S,12S,15S,18S)-1-amino-9-(2-amino-2-oxoethyl)-6-((S)-1-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)pyrrolidine-2-carboxamido)-1-imino-12-isobutyl-15-isopropyl-7,10,13,16-tetraoxo-18-(3-ureidopropyl)-2, Synthesis of 8,11,14,17-pentaazanonadecan-19-amido)benzyl((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (L013-P005)

[2192] Compound 1.3.2 (30.0 mg, 43.4 μmol, 1 eq.) and compound 1.4.5.5 (29.2 mg, 34.7 μmol, 0.8 eq.) were dissolved in DMF (0.5 mL), and PYBOP (45.1 mg, 86.7 μmol) and DIEA (30.2 μL, 173 μmol, 4 eq.) were added. The mixture was stirred at 25°C for 1 hour. LC-MS confirmed the reaction was complete. TFA was added to the reaction mixture to adjust the pH. Purification was performed by high-performance liquid chromatography. Compound L013-P005 (10.7 mg, 6.57 μmol, 15.1% yield) was obtained as a yellow solid. ESI-MS (m / z): 1514.9 [M+H] + .

[2193] 1.4.12 Synthesis of L013-P002

[2194] 1.4.12.1 4-((6S,9S,12S,15S,18S)-1-amino-9-(2-amino-2-oxoethyl)-6-((S)-1-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)pyrrolidine-2-carboxamido)-1-imino-12-isobutyl-15-isopropyl-7,10,13,16-tetraoxo-18-(3-ureidopropyl)-2,8,11,14,17-pentaazadecanoyl Synthesis of Nonadecan-19-amido)benzyl (1R,2R)-3-((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl) (methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl 1-oxobutan-2-yl (methyl)carbamate (L013-P002)

[2195] Compound 1.3.2 (30.0 mg, 43.4 μmol, 1 eq.) and compound Val-Cit-PAB-MMAE (34.1 mg, 30.4 μmol, 0.7 eq.) were dissolved in DMF (0.5 mL), and PYBOP (45.1 mg, 86.7 μmol, 2 eq.) and DIEA (30.2 μL, 173 μmol, 4 eq.) were added to the resulting solution. The mixture was stirred at 25° C. for 1 hour. LC-MS detected the completion of the reaction. TFA was added to the reaction mixture to adjust the pH. The residue was purified by preparative HPLC. Compound L013-P002 (10.7 mg, 5.60 μmol, yield: 12.9%) was obtained as a white solid. ESI-MS (m / z): 1798.6 [M+H] + .

[2196] 1.4.13 Synthesis of L015-P005

[2197] 1.4.13.1 4-((6S,9S,12S,15S,18S)-1-amino-9-(2-amino-2-oxoethyl)-6-((S)-1-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glyceryl)pyrrolidine-2-carboxamido)-1-imino-12-isobutyl-15-isopropyl-7,10,13,16-tetraoxo-18-(3-ureidopropyl)- Synthesis of 2,8,11,14,17-pentaazanonadecan-19-amido)benzyl((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13.15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (L015-P005)

[2198] Compound 1.3.3 (35.0 mg, 46.7 μmol, 1 eq.) and compound 1.4.5.5 (39.3 mg, 46.7 μmol, 1 eq.) were dissolved in DMF (0.5 mL). PYBOP (48.7 mg, 93.5 μmol, 2 eq.) and DIEA (32.6 μL, 187 μmol, 4 eq.) were added to the resulting solution, and the mixture was stirred at 25° C. for 1 hour. LC-MS confirmed the completion of the reaction. TFA was added to the reaction mixture to adjust the pH. The residue was purified by preparative HPLC. Compound L015-P005 (10.4 mg, 6.17 μmol, yield: 13.2%) was obtained as a yellow solid. ESI-MS (m / z): 1572.7 [M+H] + .

[2199] 1.4.14 Synthesis of L015-P002

[2200] 1.4.14.1 4-((6S,9S,12S,15S,18S)-1-amino-9-(2-amino-2-oxoethyl)-6-((S)-1-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glyceryl)pyrrolidine-2-carboxamido)-1-imino-12-isobutyl-15-isopropyl-7,10,13,16-tetraoxo-18-(3-ureidopropyl)-2,8,11,14,17-pentaazadecanoyl Synthesis of Nonadecan-19-amido)benzyl-2-((1R,2R)-3-((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl1-oxobutan-2-yl(methyl)carbamate (L015-P002)

[2201] Compound 1.3.3 (35.0 mg, 46.7 μmol, 1 eq) and compound Val-Cit-PAB-MMAE (42.0 mg, 37.4 μmol, 0.8 eq) were dissolved in DMF (0.5 mL), and PYBOP (48.7 mg, 93.5 μmol, 2 eq) and DIEA (24.4 μL, 140 μmol, 3 eq) were added to the resulting solution, and the mixture was stirred at 25 ° C for 1 hour. LC-MS detection showed that the reaction was complete. TFA was added to the reaction mixture to adjust the pH value. The residue was purified by preparative HPLC to give compound L015-P002 (11.4 mg, 5.79 μmol, yield 12.4%, TFA) as a white solid. ESI-MS (m / z): 1855.1 [M + H] + .

[2202] 1.4.15 Synthesis of L015-P003

[2203] 1.4.15.1 Synthesis of Fmoc-Val-Cit-PAB-ET743 (1.4.15.1)

[2204] Compound 1.4.5.3 (101 mg, 131 μmol, 2.00 eq.) and ET743 (P003) (50.0 mg, 65.6 μmol, 1.00 eq.) were dissolved in DMF (100 μL), and DIEA (45.7 μL, 263 μmol, 4.00 eq.) was added to the resulting solution. The mixture was stirred at 25°C for 8 hours. LC-MS confirmed the reaction was complete. The reaction mixture was quenched with H2O (1.00 mL) and purified by preparative HPLC to afford compound 1.4.15.1 (50.0 mg, 36.0 μmol, yield: 54.8%) as a white solid. ESI-MS (m / z): 1372.7 [M-H2O+H] + .

[2205] 1.4.15.2 Synthesis of Val-Cit-PAB-ET743 (1.4.15.2)

[2206] Compound 1.4.15.1 (50.0 mg, 36.0 μmol, 1.00 eq) was dissolved in DMF (400 μL) and TEA (100 μL) was added to the resulting solution. The mixture was stirred at 25°C for 6 hours. LC-MS confirmed the reaction was complete. The reaction mixture was filtered to remove insoluble solids and purified by prep-HPLC to afford compound 1.4.15.2 (33.0 mg, 28.3 μmol, yield: 78.6%) as a white solid. ESI-MS (m / z): 1150.8 [M-H2O+H] + .

[2207] 1.4.15.3 Synthesis of L015-P003

[2208] Compound 1.4.15.2 (33.0 mg, 28.3 μmol, 1.00 eq) was dissolved in DMF (500 μL) and PYBOP (22.1 mg, 42.4 μmol, 1.50 eq), DIEA (19.7 μL, 113 μmol, 4.00 eq) and compound 1.3.3 (25.4 mg, 33.9 μmol, 1.20 eq) were added to the resulting solution. The mixture was stirred at 25°C for 6 hours. LC-MS confirmed the reaction was complete. The reaction mixture was filtered to remove insoluble solids and purified by prep-HPLC to afford compound L015-P003 (8.60 mg, 4.53 μmol, yield: 16.0%) as a white solid. ESI-MS (m / z): 1899.1 [M+H] + .

[2209] 1.4.16 Synthesis of L016-P005

[2210] 1.4.16.1 4-((6S,9S,12S,15S,18S)-1-amino-9-(2-amino-2-oxoethyl)-12-((dimethylamino)methyl)-6-((S)-1-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glyceryl)pyrrolidine-2-carboxamido)-1-imino-15-isopropyl-7,10,13,16-tetraoxo-18-(3-ureidopropyl) Synthesis of (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (L016-P005)

[2211] Compound 1.3.1 (15.3 mg, 20.4 μmol, 1 eq) and compound 1.4.5.5 (15.4 mg, 18.4 μmol, 0.9 eq) were dissolved in DMF (0.3 mL), and PYBOP (21.2 mg, 40.8 μmol, 2 eq) and DIEA (14.2 μL, 81.6 μmol, 4 eq) were added to the solution. The mixture was stirred at 25 ° C for 0.5 hours. LC-MS detected that the reaction was complete. TFA was added to the reaction mixture to adjust the pH value. The residue was purified by preparative HPLC. Compound L016-P005 (6.60 mg, 3.81 μmol, yield: 18.7%, TFA) was obtained as a yellow solid. ESI-MS (m / z): 1573.7 [M+H] + .

[2212] 1.4.17 Synthesis of L016-P002

[2213] 1.3.17.1 4-((6S,9S,12S,15S,18S)-1-amino-9-(2-amino-2-oxoethyl)-12-((dimethylamino)methyl)-6-((S)-1-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glyceryl)pyrrolidine-2-carboxamido)-1-imino-15-isopropyl-7,10,13,16-tetraoxo-18-(3-ureidopropyl)-2,8,11,14,17-pentaazadecanoyl Synthesis of Nonadecan-19-amido)benzyl-1-((S)-2-((1R,2R)-3-((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)amino)-3-methyl1-oxobutan-2-yl(methyl)carbamate (L016-P002)

[2214] Compound 1.3.1 (15.0 mg, 20.0 μmol, 1 eq.) and compound Val-Cit-PAB-MMAE (15.7 mg, 14.0 μmol, 0.7 eq.) were dissolved in DMF (0.3 mL), and PYBOP (20.8 mg, 40.0 μmol, 2 eq.) and DIEA (13.9 μL, 80.0 μmol 4 eq.) were added to the resulting solution. The mixture was stirred at 25° C. for 0.5 h. LC-MS confirmed the completion of the reaction. TFA was added to the reaction mixture to adjust the pH. The residue was purified by preparative HPLC to give compound L016-P002 (6.00 mg, 2.94 μmol, 14.7% yield, TFA) as a white solid. ESI-MS (m / z): 1856.1 [M+H] + .

[2215] 1.4.18 Synthesis of L017-P005

[2216] 1.4.18.1 4-((6S,9S,12S,15S,18S)-1-amino-9-(2-amino-2-oxoethyl)-6-((S)-1-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)-L-seryl)pyrrolidine-2-carboxamido)-1-imino-12-isobutyl-15-isopropyl-7,10,13,16-tetraoxo-18-(3-ureidopropyl) Synthesis of (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-2,8,11,14,17-pentaazanonadecan-19-amido)benzyl((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (L017-P005)

[2217] Compound 1.3.4 (30.0 mg, 38.5 μmol, 1 eq.) and compound 1.4.5.5 (32.4 mg, 38.5 μmol, 1 eq.) were dissolved in DMF (0.5 mL), and PYBOP (40.1 mg, 77.0 μmol, 2 eq.) and DIEA (20.1 μL, 116 μmol, 3 eq.) were added to the resulting solution. The mixture was stirred at 25°C for 1 hour. LC-MS confirmed the completion of the reaction. TFA was added to the reaction mixture to adjust the pH. The residue was purified by preparative HPLC. Compound L017-P005 (11.0 mg, 6.11 μmol, yield: 15.9%, TFA) was obtained as a yellow solid. ESI-MS (m / z): 1602.5 [M+H] + .

[2218] 1.4.19 Synthesis of L017-P002

[2219] 1.4.19.1 4-((6S,9S,12S,15S,18S)-1-amino-9-(2-amino-2-oxoethyl)-6-((S)-1-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)-L-acyl)pyrrolidine-2-carboxamido)-1-imino-12-isobutyl-15-isopropyl-7,10,13,16-tetraoxo-18-(3-ureidopropyl)-2,8,11,14,17-pentaazanonadecane Synthesis of (S)-2-((1R,2R)-3-((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptane-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamate (L017-P002)

[2220] Compound 1.3.4 (30.0 mg, 38.5 μmol, 1 eq) and compound Val-Cit-PAB-MMAE (34.6 mg, 30.8 μmol, 0.8 eq) were dissolved in DMF (0.5 mL), and PYBOP (40.1 mg, 77.0 μmol, 2 eq) and DIEA (26.8 μL, 154 μmol, 4 eq) were added to the resulting solution. The mixture was stirred at 25 ° C for 1 hour. LC-MS detection showed that the reaction was complete. TFA was added to the reaction mixture to adjust the pH value. The residue was purified by preparative HPLC. White solid compound L017-P002 (10.5 mg, 5.25 μmol, yield: 13.6%, TFA) was obtained. ESI-MS (m / z): 1885.3 [M+H] + .

[2221] 1.4.20 Synthesis of L021-P005

[2222] 1.4.20.1 4-((3S,6S,9S,12S,15S)-1-((S)-1-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glyceryl)pyrrolidin-2-yl)-6-(hydroxymethyl)-3,9-diisobutyl-12-isopropyl-1,4,7,10,13-pentaoxo-15-(3-ureidopropyl)-2,5,8,11, Synthesis of 14-pentaazahexadecane-16-amido)benzyl((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (L021-P005)

[2223] Compound 1.3.5 (55.0 mg, 81.0 μmol, 1.00 eq.) and compound 1.4.5.5 (68.1 mg, 81.1 μmol, 1.00 eq.) were dissolved in DMF (0.50 mL). PYBOP (84.3 mg, 162 μmol, 2.00 eq) and DIEA (41.9 mg, 324 μmol, 56.5 μL, 4.00 eq) were added to the resulting solution. The mixture was stirred at 25°C for 1 hour. LC-MS confirmed the completion of the reaction. The residue was purified by preparative HPLC to afford L021-P005 (25.0 mg, 16.4 μmol, yield: 20.3%, TFA) as a yellow solid. ESI-MS (m / z): 1502.8 [M+H] + .

[2224] 1.4.21 Synthesis of L021-P002

[2225] 1.4.21.1 4-((3S,6S,9S,12S,15S)-1-((S)-1-((6-(2,5-dioxy-2,5-dihydro-1H-pyrrol-1-yl)hexyl)glycylamino)pyrrolidin-2-yl)-6-(hydroxymethyl)-3,9-diisobutyl-12-isopropyl-1,4,7,10,13-pentaoxy-15-(3-ureidopropyl)-2,5,8,11,14-pentaazahexadecane-16-amido)benzyl ((S)-1-((S)-1-)( Synthesis of (3R,4S,5S)-1-((S)-2-((1R,2R)-3-((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxohept-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamate (L021-P002)

[2226] To a solution of compound 1.3.5 (50.0 mg, 73.7 μmol, 1.00 eq.) and compound Val-Cit-PAB-MMAE (82.8 mg, 73.7 μmol, 1.00 eq.) in DMF (0.50 mL) were added PYBOP (76.7 mg, 147 μmol, 2.00 eq.) and DIEA (38.1 mg, 295 μmol, 51.3 μL, 4.00 eq.). The mixture was stirred at 25°C for 1 hour. LC-MS confirmed the reaction was complete. Purification by preparative HPLC afforded L021-P002 (44.0 mg, 24.3 μmol, 33.0% yield, TFA) as a white solid. ESI-MS (m / z): 1784.2 [M+H] + .

[2227] 1.4.22 Synthesis of L027-P005

[2228] 1.4.22.1 4-((6S,12S,15S,18S)-1-amino-6-((S)-1-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)pyrrolidine-2-carboxamido)-1-imino-12-isobutyl-15-isopropyl-7,10,13,16-tetraoxo-18-(3-ureidopropyl)-2,8,11,14,1 Synthesis of 7-pentaazanonadecan-19-amido)benzyl((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (L027-P005)

[2229] Compound 1.4.5.5 (100 mg, 118 μmol, 1.0 eq.) and compound 1.3.6 (75.4 mg, 118 μmol, 1.0 eq.) were dissolved in DMF (0.20 mL), and TBTU (45.8 mg, 142 μmol, 1.2 eq.) and DIEA (46.1 mg, 356 μmol, 62.1 μL, 3.0 eq.) were added to the resulting solution. The mixture was stirred at 25° C. for 1.0 hour. LC-MS detected the reaction to be complete. The mixture was purified twice by preparative HPLC to give L027-P005 (45.0 mg, 30.8 μmol, yield: 25.9%) as a yellow solid. ESI-MS (m / z): 1458.6 [M+H] + .

[2230] 1.4.23 Synthesis of L028-P005

[2231] 1.4.23.1 Synthesis of 4-((S)-2-((S)-2-((S)-2-((S)-1-((6-(2,5-dioxy-2,5-dihydro-1H-pyrrol-1-yl)hexyl)-L-prolyl-L-arginyl)pyrrolidine-2-carboxamido)-4-methylpentaamido)-3-methylbutanamido)-5-ureidopentaamido)benzyl((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxy-2,3,9,10,13-dioxyhexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indeno[1,2-b]quinolin-1-yl)carbamate (L028-P005)

[2232] Compound 1.4.5.5 (100 mg, 118 μmol, 1.0 eq.) and compound 1.3.7 (80.2 mg, 118 μmol, 1.0 eq.) were dissolved in DMF (0.20 mL). To the resulting solution were added TBTU (45.8 mg, 142 μmol, 1.2 eq.) and DIEA (46.1 mg, 356 μmol, 62.1 μL, 3.0 eq.). The mixture was stirred at 25°C for 1.0 hour. LC-MS analysis indicated that the reaction was complete. The mixture was purified by preparative HPLC to afford L028-P005 (36.0 mg, 24.0 μmol, 20.2% yield) as a yellow solid. ESI-MS (m / z): 1498.0 [M+H] + .

[2233] 1.4.24 Synthesis of L029-P005

[2234] 1.4.24.1 4-((6S,9S,12S,15S,18S)-1-amino-6-((S)-1-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glyceryl)pyrrolidine-2-carboxamido)-9-(hydroxymethyl)-1-imino-12-isobutyl-15-isopropyl-7,10,13,16-tetraoxo-18-(3-ureidopropyl)-2,8, Synthesis of 11,14,17-pentaazanonadecan-19-amido)benzyl((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (L029-P005)

[2235] Compound 1.4.5.5 (100 mg, 118 μmol, 1.0 eq.) and compound 1.3.8 (85.8 mg, 118 μmol, 1.0 eq.) were dissolved in DMF (0.20 mL), and TBTU (45.8 mg, 142 μmol, 1.2 eq.) and DIEA (46.1 mg, 356 μmol, 62.1 μL, 3.0 eq.) were added to the resulting solution. The mixture was stirred at 25°C for 1.0 hour. LC-MS detected the reaction to be complete. The mixture was purified by preparative HPLC to give L029-P005 (65.0 mg, 42.0 μmol, yield: 35.3%) as a yellow solid. ESI-MS (m / z): 1545.5 [M+H] + .

[2236] 1.4.25 Synthesis of L030-P005

[2237] 1.4.25.1 Synthesis of 4-((S)-2-((S)-2-((S)-2-((S)-1-((6-(2,5-dioxy-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycyl-L-prolyl-L-arginyl)pyrrolidine-2-carboxamido)-4-methylpentaamido)-3-methylbutylamino)-5-ureidopentaamido)benzyl((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxy-2,3,9,10,13-dioxy,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizin[1,2-b]quinolin-1-yl)carbamate (L030-P005)

[2238] TBTU (27.4 mg, 85.6 μmol, 1.2 eq.) and DIEA (18.4 mg, 142 μmol, 24.8 μL, 2.0 eq.) were added to a solution of compound 1.3.9 (47.0 mg, 64.2 μmol, 0.9 eq.) and compound 1.4.5.5 (60.0 mg, 71.33 μmol, 1.0 eq.) in DMF (1.00 mL). The mixture was stirred at 25° C. for 1.0 hour. LC-MS detected the reaction to be complete. The mixture was purified twice by preparative HPLC to give L030-P005 (24.0 mg, 15.4 μmol, yield: 21.6%) as a yellow solid. ESI-MS (m / z): 1554.7 [M+H] + .

[2239] 1.4.26 Synthesis of L031-P005

[2240] 1.4.26.1 4-((S)-2-((S)-2-((S)-2-((S)-1-((S)-2-((S)-1-((6-(2,5-dioxy-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycylamino)pyrrolidine-2-carboxamido)-5-ureidopentanyl)pyrrolidine-2-carboxamido)-4-methylpentanamido)-3-methylbutanamido Synthesis of (5-ureidopentanamido)benzyl ((1S,9S)-9-ethyl-5-fluoro-9-hydroxy)-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolo[1,2-b]quinolin-1-yl)carbamate (L031-P005)

[2241] Compound 1.4.5.5 (60.0 mg, 71.3 μmol, 1.0 eq.) and compound 1.3.10 (47.0 mg, 64.2 μmol, 0.9 eq.) were dissolved in DMF (1.00 mL). To the resulting solution were added TBTU (25.2 mg, 78.4 μmol, 1.1 eq.) and DIEA (18.4 mg, 142 μmol, 24.8 μL, 2.0 eq.). The mixture was stirred at 25°C for 1.0 hour. LC-MS analysis indicated that the reaction was complete. The mixture was purified by preparative HPLC to afford L031-P005 (14.0 mg, 9.00 μmol, 12.6% yield) as a yellow solid. ESI-MS (m / z): 1556.7 [M+H] + .

[2242] 1.4.27 Synthesis of L032-P005

[2243] 1.4.27.1 4-((6S,12S,15S,18S)-6-((S)-1-(Na-(6-(2,5-dioxy-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)-Nt-triphenyl-L-histidyl)pyrrolidine-2-carboxamido)-1-imino-12-isobutyl-15-isopropyl-7,10,13,16-tetraoxy-1-((2,2,4,6,7-pentamethyl-2,3-dihydrobenzofuran)-5-sulfonyl ... Synthesis of (1.4.27.1)-18-(3-ureidopropyl)-2,8,11,14,17-pentaazanonane-19-amido)benzyl((1S,9S)-...

Claims

1. A polypeptide or polypeptide fragment having a structure represented by -P1-P2-, wherein: P1 comprises a polypeptide responsive to matrix metalloproteinase 2, matrix metalloproteinase 9 or cathepsin B, and P2 comprises a polypeptide responsive to cathepsin B, matrix metalloproteinase 2 or matrix metalloproteinase 9.

2. The polypeptide or polypeptide fragment according to claim 1, wherein The P1 comprises the following polypeptide structure: -X1-P-X2-X3-Z-, and the P2 comprises the following polypeptide structure: -X1'-X2'-X3'-X4'-; wherein P is a proline residue, X1 represents absence, a single amino acid or a single amino acid derivative, X2, X3, and Z represent a single amino acid or a single amino acid derivative, X1' and X2' represent a single amino acid or a single amino acid derivative, and X3' and X4' represent absence or a single amino acid or a single amino acid derivative.

3. The polypeptide or polypeptide fragment according to claim 2, wherein: The X1 is selected from: absent, Gly (G), Val (V), His (H), Ser (S), Ile (I); The X2 is selected from: Ala (A), Ser (S), Leu (L), Arg (R), Gln (Q), Val (V), Cit; The X3 is selected from: Gly (G), Ser (S), Pro (P), Asn (N), Ala (A); The Z is selected from: Leu (L), Ile (I), Val (V), Phe (F), Met (M), Ala (A), Trp (W), Gln (Q), N,N-dimethyl-3-aminopropionic acid (Dap(Me2)), N,N-dimethyl-4-aminobutyric acid (Dab(Me2)), N,N-dimethylornithine (Orn(Me2)), N,N-dimethyllysine (Lys(Me2)), N,N-diethyl-3-aminopropionic acid (Dap(Et2)), N,N-diethyl 4-aminobutyric acid (Dab(Et2)), N,N-diethylornithine (Orn(Et2)), N,N-diethyllysine (Lys(Et2)).

4. The polypeptide or polypeptide fragment according to claim 2 or 3, wherein: The X1' is selected from the group consisting of: Val (V), Als (A), Leu (L), Ile (I), Met (M), Phe (F), Trp (W), Pro (P), Tyr (Y), Gly (G), Glu (E); The X2' is selected from: Val (V), Leu (L), Ile (I), Met (M), Trp (W), Pro (P), Lys (K), Lys (Me2), Lys (Et2), His (H), Ala (A), Arg (R), Gly (G), Phe (F), Cit, Orn, Orn (Me2), Orn (Et2); The X3' is selected from: absent, Phe (F), Gly (G), Lys (K), Lys (Me2), Lys (Et2), His (H), Ala (A), Arg (R), Cit, Orn, Orn (Me2), Orn (Et2); The X4' is selected from: absent, Gly (G), Phe (F).

5. The polypeptide or polypeptide fragment according to any one of claims 2 to 4, wherein: The X1 is selected from: absent, Gly (G), Val (V), His (H), Ser (S), Ile (I); The X2 is selected from: Ala (A), Ser (S), Leu (L), Arg (R), Gln (Q), Val (V), Cit; The X3 is selected from: Gly (G), Ser (S), Pro (P), Asn (N), Ala (A); The Z is selected from the group consisting of Leu (L), Ile (I), Val (V), Phe (F), Met (M), Ala (A), Trp (W), Gln (Q), N,N-dimethyl-3-aminopropionic acid (Dap(Me2)), N,N-dimethyl-4-aminobutyric acid (Dab(Me2)), N,N-dimethylornithine (Orn(Me2)), N,N-dimethyllysine (Lys(Me2)), N,N-diethyl-3-aminopropionic acid (Dap(Et2)), N,N-diethyl-4-aminobutyric acid (Dab(Et2)), N,N-diethylornithine (Orn(Et2)), N,N-diethyllysine (Lys(Et2)); The X1' is selected from the group consisting of: Val (V), Als (A), Leu (L), Ile (I), Met (M), Phe (F), Trp (W), Pro (P), Tyr (Y), Gly (G), Glu (E); The X2' is selected from: Val (V), Leu (L), Ile (I), Met (M), Trp (W), Pro (P), Lys (K), Lys (Me2), Lys (Et2), His (H), Ala (A), Arg (R), Gly (G), Phe (F), Cit, Orn, Orn (Me2), Orn (Et2); The X3' is selected from: absent, Phe (F), Gly (G), Lys (K), Lys (Me2), Lys (Et2), His (H), Ala (A), Arg (R), Cit, Orn, Orn (Me2), Orn (Et2); The X4' is selected from: absent, Gly (G), Phe (F); Preferably, X1 is absent or is Gly (G), Ser (S) or His (H); X2 is Ala (A), Leu (L), Arg (R), Ser (S), Leu (L) or Cit; X3 is Gly (G), Ser (S), Ala (A), Asn (N) or Pro (P); Z is Leu (L) or Dap (Me2); X1' is Val (V) or Gly (G), X2' is Gly (G) or Cit, X3' and X4' are absent at the same time, or X3' is Phe (F) and X4' is Gly (G).

6. The polypeptide or polypeptide fragment according to any one of claims 1 to 5, wherein: The P2 is -Val-Cit-, -Ala-Lys- or -Gly-Gly-Phe-Gly-.

7. The polypeptide or polypeptide fragment according to any one of claims 1 to 6, which is selected from: -His-Val-Leu-Asn-Leu-Val-Cit-(SEQ ID NO: 1); -Val-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:2); -Ile-Pro-Val-Ser-Leu-Val-Cit-(SEQ ID NO:3); -Gly-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:4); -Gly-Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:5); -Gly-Pro-Ala-Gly-Leu-Val-Cit-(SEQ ID NO: 6); -Gly-Pro-Gln-Gly-Leu-Val-Cit-(SEQ ID NO:7); -Gly-Pro-Leu-Gly-Leu-Val-Cit-(SEQ ID NO:8); -Gly-Pro-Ser-Gly-Leu-Val-Cit-(SEQ ID NO:9); -Gly-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO: 10); -Gly-Pro-Ala-Ala-Leu-Val-Cit-(SEQ ID NO: 11); -Gly-Pro-Ser-Ala-Leu-Val-Cit-(SEQ ID NO:12); -His-Val-Leu-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO: 13); -Val-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO: 14); -Ile-Pro-Val-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO: 15); -Gly-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO: 16); -Gly-Pro-Ala-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO: 17); -Gly-Pro-Ala-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO: 18); -Gly-Pro-Gln-Gly-Leu-Gly-Gly-Phe-Gly- (SEQ ID NO: 19); -Gly-Pro-Leu-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO: 20); -Gly-Pro-Ser-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:21); -Gly-Pro-Arg-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:22); -Gly-Pro-Ala-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:23); -Gly-Pro-Ser-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:24); -Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:25); -Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:26); -Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:27); -Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:28); -Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:29); -Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:30); -Gly-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:31); -Gly-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:32); -Gly-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:33); -Gly-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:34); -Gly-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:35); -His-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:36); -His-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:37); -His-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:38); -His-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:39); -His-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:40); -His-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:41); -Ser-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:42); -Ser-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:43); -Ser-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:44); -Ser-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:45); -Ser-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:46); -Ser-Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:47); -Ser-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:48); -Gly-Pro-Ala-Pro-Leu-Val-Cit-(SEQ ID NO:49); -Gly-Pro-Ala-Asn-Leu-Val-Cit-(SEQ ID NO:50); -Gly-Pro-Arg-Gly-Dap(Me2)-Val-Cit-(SEQ ID NO:51); -Gly-Pro-Arg-Asn-Dap(Me2)-Val-Cit-(SEQ ID NO:60); -Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:61); -Gly-Pro-Arg-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO: 62); -Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:63); -Gly-Pro-Cit-Asn-Leu-Val-Cit-(SEQ ID NO: 64); -Ser-Pro-Cit-Ser-Dap(Me2)-Val-Cit-(SEQ ID NO:65); -Pro-Arg-Ser-Dap(Me2)-Val-Cit-(SEQ ID NO:66); -Pro-Cit-Asn-Leu-Val-Cit- (SEQ ID NO: 67); and -Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:68); or a variant having one or two mismatches relative to the sequence of any one of SEQ ID NOs: 1-51 and 60-68; Preferably, the polypeptide or polypeptide fragment is selected from: -GPLSLVCit-(SEQ ID NO:4); -GPASLVCit- (SEQ ID NO: 5); -GPAGLVCit- (SEQ ID NO: 6); -GPRGLVCit- (SEQ ID NO: 10); -GPSALVCit-(SEQ ID NO: 12); -GPLGLGGFG- (SEQ ID NO: 20); -GPRGLGGFG- (SEQ ID NO: 22); -GPAALGGFG- (SEQ ID NO: 23); -PRGLVCit-(SEQ ID NO:25); -PRSLVCit- (SEQ ID NO: 26); -PRNLVCit- (SEQ ID NO: 27); -PRPLVCit- (SEQ ID NO: 28); -PCitGLVCit- (SEQ ID NO: 29); -PCitPLVCit- (SEQ ID NO: 30); -GPRSLVCit- (SEQ ID NO: 31); -GPRPLVCit- (SEQ ID NO: 32); -GPRNLVCit- (SEQ ID NO: 33); -GPCitGLVCit-(SEQ ID NO:34); -GPCitPLVCit-(SEQ ID NO:35); -HPRGLVCit- (SEQ ID NO: 36); -HPCitPLVCit-(SEQ ID NO:41); -SPRGLVCit- (SEQ ID NO: 42); -SPRSLVCit- (SEQ ID NO: 43); -SPRPLVCit- (SEQ ID NO: 44); -SPRNLVCit- (SEQ ID NO: 45); -SPCitGLVCit-(SEQ ID NO:46); -SPCitSLVCit-(SEQ ID NO:47); -SPCitPLVCit-(SEQ ID NO:48); -GPAPLVCit- (SEQ ID NO: 49); -GPANLVCit- (SEQ ID NO: 50); -GPRNDap(Me2)VCit-(SEQ ID NO:60); -PLSLVCit- (SEQ ID NO: 61); -GPRNLGGFG- (SEQ ID NO: 62); -PCitSLVCit- (SEQ ID NO: 63); -GPCitNLVCit-(SEQ ID NO:64); -SPCitSDap(Me2)VCit-(SEQ ID NO:65); -PRSDap(Me2)VCit-(SEQ ID NO:66); -PCitNLVCit- (SEQ ID NO: 67); and -PASLVCit- (SEQ ID NO: 68).

8. The polypeptide or polypeptide fragment according to any one of claims 1 to 7, selected from: -Gly-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:4); -Gly-Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:5); -Gly-Pro-Ala-Gly-Leu-Val-Cit-(SEQ ID NO:6); -Gly-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:10); -Gly-Pro-Ser-Ala-Leu-Val-Cit-(SEQ ID NO:12); -Gly-Pro-Leu-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:20); -Gly-Pro-Arg-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:22); -Gly-Pro-Ala-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:23); -Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:25); -Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:26); -Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:27); -Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:28); -Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:29); -Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:30); -Gly-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:31); -Gly-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:32); -Gly-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:33); -Gly-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:34); -Gly-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:35); -His-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:36); -Ser-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:42); -Ser-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:43); -Ser-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:44); -Ser-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:45); -Ser-Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:47); -Ser-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:48); -Gly-Pro-Ala-Asn-Leu-Val-Cit-(SEQ ID NO:50); -Gly-Pro-Arg-Asn-Dap(Me2)-Val-Cit-(SEQ ID NO:60); -Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:61); -Gly-Pro-Arg-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO: 62); -Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:63); -Gly-Pro-Cit-Asn-Leu-Val-Cit-(SEQ ID NO: 64); -Ser-Pro-Cit-Ser-Dap(Me2)-Val-Cit-(SEQ ID NO:65); -Pro-Arg-Ser-Dap(Me2)-Val-Cit-(SEQ ID NO:66); -Pro-Cit-Asn-Leu-Val-Cit- (SEQ ID NO: 67); and -Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:68); Preferably, the polypeptide or polypeptide fragment is one or more selected from the following: -GPASLVCit- (SEQ ID NO: 5); -GPAGLVCit- (SEQ ID NO: 6); -GPRGLVCit- (SEQ ID NO: 10); -GPSALVCit-(SEQ ID NO: 12); -GPLGLGGFG- (SEQ ID NO: 20); -GPRGLGGFG- (SEQ ID NO: 22); -GPAALGGFG- (SEQ ID NO: 23); -PRGLVCit-(SEQ ID NO:25); -PRSLVCit- (SEQ ID NO: 26); -PRNLVCit- (SEQ ID NO: 27); -PRPLVCit- (SEQ ID NO: 28); -PCitGLVCit- (SEQ ID NO: 29); -PCitPLVCit- (SEQ ID NO: 30); -GPRSLVCit- (SEQ ID NO: 31); -GPRPLVCit- (SEQ ID NO: 32); -GPRNLVCit- (SEQ ID NO: 33); -GPCitGLVCit-(SEQ ID NO:34); -GPCitPLVCit-(SEQ ID NO:35); -HPRGLVCit- (SEQ ID NO: 36); -HPCitPLVCit-(SEQ ID NO:41); -SPRGLVCit- (SEQ ID NO: 42); -SPRSLVCit- (SEQ ID NO: 43); -SPRPLVCit- (SEQ ID NO: 44); -SPRNLVCit- (SEQ ID NO: 45); -SPCitGLVCit-(SEQ ID NO:46); -SPCitSLVCit-(SEQ ID NO:47); -SPCitPLVCit-(SEQ ID NO:48); -GPAPLVCit- (SEQ ID NO: 49); -GPANLVCit- (SEQ ID NO: 50); -GPRNDap(Me2)VCit-(SEQ ID NO:60); -PLSLVCit- (SEQ ID NO: 61); -GPRNLGGFG- (SEQ ID NO: 62); -PCitSLVCit- (SEQ ID NO: 63); -GPCitNLVCit-(SEQ ID NO:64); -SPCitSDap(Me2)VCit-(SEQ ID NO:65); -PRSDap(Me2)VCit-(SEQ ID NO:66); -PCitNLVCit- (SEQ ID NO: 67); -PASLVCit- (SEQ ID NO: 68).

9. A linker comprising the polypeptide or polypeptide fragment having the structure represented by -P1-P2- according to any one of claims 1 to 8, wherein: The linker has the following general formula: C1-La-L1-P1-P2-L2; Where C1 is the connector; La is empty or a tuning spacer; L1 is empty or a spacer; L2 is empty or a spacer; The positions of La and L1 can be swapped.

10. The linker according to claim 9, wherein C1 is selected from the following structures: in Indicates the connection position with the biological ligand unit, Indicates the connection position with La.

11. The linker according to claim 9 or 10, wherein The C1 is selected from the following structures: -(succinimide-3-yl-N)-, -CH2-C(=O)-, -C(=O)- or Wherein, the -(succinimide-3-yl-N)- has the following structure: in Indicates the connection position with the biological ligand unit, Indicates the connection position with La.

12. The linker according to any one of claims 9 to 11, wherein The La is selected from the following structures or a combination of the following structures: R1-SS-R2, R1-(CH2CH2O) m -R2, R1-(CH2CH2O) m -CH2CH2S-(CH2CH2O) n -R2, R1-(CH2CH2O) m -CH2CH2NH-(CH2CH2O) n -R2, R1-(CH2CH2O) m -CH2CH2(C=O)NH-(CH2CH2O) n -R2, a is 0 or 1; m is any integer from 1 to 20; n is any integer from 1 to 20; R3 is H, -C1-C 10 Alkylene-, -C3-C8 carbocyclic group-, -C2-C 10 Heteroaryl-, -C3-C 24 Alkylheteroaryl-, wherein -C1-C 10 Alkylene-, -C3-C8 carbocyclic group-, -C2-C 10 Heteroaryl-, -C3-C 24 Alkylheteroaryl- may be substituted by one or more heteroatoms, which may be O, S, NR 3 , where R 3 is H or -C1-C4 alkylene; R1 is a connection structure with C1, and R2 is a connection structure with L1; The R1 is a single bond or is selected from the following structures: -X 1 -C1-C 10 Alkylene-X 2 -、-X 1 -C1-C 10 Heteroalkylene-X 2 -、-X 1 -C3-C8 carbocyclyl-X 2 -、-X 1 -Arylene-X 2 -、-X 1 -C1-C 10 Alkylene-arylene-X 2 -、-X 1 -Arylene-C1-C 10 Alkylene-X 2 -、-X 1 -C1-C 10 Alkylene-(C3-C8 carbocyclic group)-X 2 -、-X 1 -(C3-C8 carbocyclyl)-C1-C 10 Alkylene-X 2 -、-X 1 -C3-C8 heterocyclyl-X 2 -、-X 1 -C1-C 10 Alkylene-(C3-C8 heterocyclyl)-X 2 -、-X 1 -(C3-C8 heterocyclyl)-C1-C 10 Alkylene-X 2 -; Wherein X 1 , X 2 Each independently selected from not existing or selected from O, S, NH, C(=O), NHC(=O), C(=O)NH, NHC(=O)NH; wherein when the R1 is not a single bond, it is optionally substituted by a basic unit (BU), the basic unit is -(CH2) x NH2, -(CH2) x NHR m or -(CH2) x N(R m )2; where x is any integer from 1 to 4; and each R m are independently selected from C1-C6 alkyl and C1-C6 haloalkyl, or two R m The groups combine with the nitrogen to which they are attached to form a 4-6 membered heterocycloalkyl ring, azetidinyl, pyrrolidinyl or piperidinyl group; The R2 is a single bond or is selected from the following structures: -X 3 -C1-C 10 Alkylene-X 4 -、-X 3 -C1-C 10 Heteroalkylene-X 4 -、-X 3 -C3-C8 carbocyclyl-X 4 -、-X 3 -Arylene-X 4 -、-X 3 -C1-C 10 Alkylene-arylene-X 4 -、-X 3 -Arylene-C1-C 10 Alkylene-X 4 -、-X 3 -C1-C 10 Alkylene-(C3-C8 carbocyclic group)-X 4 -、-X 3 -(C3-C8 carbocyclyl)-C1-C 10 Alkylene-X 4 -、-X 3 -C3-C8 heterocyclyl-X 4 -、-X 3 -C1-C 10 Alkylene-(C3-C8 heterocyclyl)-X 4 -、-X 3 -(C3-C8 heterocyclyl)-C1-C 10 Alkylene-X 4 -; Wherein X 3 is absent or selected from O, S, NH, C(=O), NHC(=O), C(=O)NH, NHC(=O)NH, X 4 is absent or selected from O, NH, S, C(=O), C(=O)NH, NHC(=O), NHC(=O)NH; wherein when the R2 is not a single bond, it is optionally substituted by a basic unit (BU), the basic unit being -(CH2) x NH2, -(CH2) x NHR m or -(CH2) x N(R m )2; where x is any integer from 1 to 4; and each R m are independently selected from C1-C6 alkyl and C1-C6 haloalkyl, or two R m The groups combine with the nitrogen to which they are attached to form a 4-6 membered heterocycloalkyl ring, azetidinyl, pyrrolidinyl or piperidinyl group.

13. The linker according to any one of claims 9 to 12, wherein The L1 is selected from the following structures: -X 5 -C1-C 10 Alkylene-X 6 -、-X 5 -C1-C 10 Heteroalkylene-X 6 -、-X 5 -C3-C8 carbocyclyl-X 6 -、-X 5 -Arylene-X 6 -、-X 5 -Heteroarylene-X 6 -、-X 5 -C1-C 10 Alkylene-arylene-X 6 -、-X 5 -C1-C 10 Alkylene-heteroarylene-X 6 -、-X 5 -Arylene-C1-C 10 Alkylene-X 6 -、-X 5 -heteroarylene-C1-C 10 Alkylene-X 6 -、-X 5 -C1-C 10 Alkylene-(C3-C8 carbocyclic group)-X 6 -、-X 5 -(C3-C8 carbocyclyl)-C1-C 10 Alkylene-X 6 -、-X 5 -C3-C8 heterocyclyl-X 6 -、-X 5 -C1-C 10 Alkylene-(C3-C8 heterocyclyl)-X 6 -、-X 5 -(C3-C8 heterocyclyl)-C1-C 10 Alkylene-X 6 -、-X 5 -C3-C8 heterocyclyl-arylene-X 6 -、-X 5 -Arylene-C3-C8 heterocyclyl-X 6 -、-X 5 -C3-C8 heterocyclyl-heteroarylene-X 6 -、-X 5 -heteroarylene-C3-C8heterocyclyl-X 6 -; Wherein X 5 is absent or selected from O, S, NH, C(=O), NHC(=O), C(=O)NH, NHC(=O)NH, X 6 is absent or selected from O, NH, S, C(═O), NHC(═O), C(═O)NH, NHC(═O)NH; Wherein, any methylene unit of L1 can be independently replaced by -O-, -S-, -N(R n )-、-CH(N(R n )2)-、-CH(P(R n )2)-、-C(N(R n )2)2-、-C(OR n )(N(R n )2)-, -C(=O)-, -OC(=O)-, -C(=O)O-, -N(R n )C(=O)-、-C(=O)N(R n )-、-SO-、-SO2-、-N(R n )SO2-、-SO2N(R n )-、-P(R n )-、-P(=O)(R n )-, -P(=O)(O)-, -C(=S)-, -C(=NR n )-、-SS-、-N=N-、-N(R n )-N=、=NN(R n )-, -C=N- or -N=C-; wherein each R n are independently selected from H, C1-C6 alkyl, C3-C8 carbocyclyl, C1-C6 haloalkyl, or two R n The groups combine with the nitrogen or phosphorus to which they are attached to form a 4-6 membered heterocycloalkyl group, or an azetidinyl, pyrrolidinyl or piperidinyl group.

14. The linker according to any one of claims 9 to 13, wherein The C1-La-L1- 15. The linker according to any one of claims 9 to 14, wherein The L2 is empty or selected from the following structures: in Indicates the position of connection with the polypeptide, Indicates the location of the payload connection with a specific biological function; m is any integer from 1 to 20; n is any integer from 1 to 6; R b Independently selected from H, C1-C6 alkyl, C3-C8 carbocyclyl, C1-C6 haloalkyl, C3-C8 heterocyclyl, C1-C6 alkyl-C3-C8 heterocyclyl, substituted aryl, substituted heteroaryl, C1-C6 alkyl-substituted aryl, C1-C6 alkyl-substituted heteroaryl; Wherein any methylene unit of L2 can be independently replaced by -CHX-, -C(X2)-, -C3-C8 carbocyclyl-, -C3-C8 heterocyclyl-, -arylene-, -heteroarylene-, -O-, -S-, -N(R n )-、-CH(N(R n )2)-、-CH(P(R n )2)-、-C(N(R n )2)2-、-C(OR n )(N(R n )2)-, -C(=O)-, -OC(=O)-, -C(=O)O-, -N(R n )C(=O)-、-C(=O)N(R n )-、-SO-、-SO2-、-N(R n )SO2-、-SO2N(R n )-、-P(R n )-、-P(=O)(R n )-, -P(=O)(O)-, -C(=S)-, -C(=NR n )-、-SS-、-N=N-、-N(R n )-N=、=NN(R n )-, -C=N- or -N=C-; wherein X represents halogen or deuterium, each R n are independently selected from H, C1-C6 alkyl, C3-C8 carbocyclyl, C1-C6 haloalkyl, or two R n The groups combine with the nitrogen or phosphorus to which they are attached to form a 4-6 membered heterocycloalkyl ring, or an azetidinyl, pyrrolidinyl or piperidinyl group.

16. The linker according to any one of claims 9 to 15, wherein The L2 is empty or selected from the following structures: in Indicates the position of connection with the polypeptide, Indicates the location of the payload connection with a specific biological function; W and Y are each independently N or CR Y , and each R Y are independently H or C1-C 10 alkyl; m is any integer from 0 to 20; X is halogen or deuterium.

17. The linker according to any one of claims 9 to 16, wherein The L2 is empty or selected from: n is any integer selected from 1 to 6, 1-5, 1-4 or 1-3; Preferably, L2 is empty or selected from -PAB-, -PAB-DMEDA- and -NMEDA-.

18. The linker according to any one of claims 9 to 17, wherein The linker is selected from: -(Succinimidyl-3-yl-N)-CH2-C(=O)-P1-P2-; -(Succinimidyl-3-yl-N)-CH2CH2-C(=O)-P1-P2-; -(Succinimidyl-3-yl-N)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-; -(Succinimidyl-3-yl-N)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-; -(Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-; -(Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-; -(Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-; -(Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-; -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-; -(Succinimidyl-3-yl-N)-CH2-C(=O)-P1-P2-PAB-; -(Succinimidyl-3-yl-N)-CH2CH2-C(=O)-P1-P2-PAB-; -(Succinimidyl-3-yl-N)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-; -(Succinimidyl-3-yl-N)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-; -(Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-; -(Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-; -(Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-; -(Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-; -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-PAB-; in, Indicates the connection position with the biological ligand unit.

19. The linker according to any one of claims 9 to 18, wherein The linker is selected from: -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)-P1-P2-; -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)NH-P1-P2-; -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)NH-(CH2CH2O) m -P1-P2-; -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)NH-(CH2CH2O) m -C1-C10 alkylene-C(=O)-P1-P2-; -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)-P1-P2-PAB-; -(Succinimidyl-3-yl-N)-C1-C10 alkylene-C(=O)NH-P1-P2-PAB-; -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)NH-(CH2CH2O) m -P1-P2-PAB-; -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)NH-(CH2CH2O) m -C1-C10 alkylene-C(=O)-P1-P2-PAB-; -(Succinimidyl-3-yl-N)-C1-C10 alkylene-C(=O)-P1-P2-PAB-DMEDA-; -(Succinimidyl-3-yl-N)-C1-C10 alkylene-C(=O)NH-P1-P2-PAB-DMEDA-; -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)NH-(CH2CH2O) m -P1-P2-PAB-DMEDA-; -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)NH-(CH2CH2O) m -C1-C10 alkylene-C(=O)-P1-P2-PAB-DMEDA-; -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)-P1-P2-NMEDA-; -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)NH-P1-P2-NMEDA-; -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)NH-(CH2CH2O) m -P1-P2-NMEDA-; -(Succinimidyl-3-yl-N)-C1-C10alkylene-C(=O)NH-(CH2CH2O) m -C1-C10 alkylene-C(=O)-P1-P2-NMEDA-; -(C=O)-C1-C10 alkylene-C(=O)-P1-P2-; -(C=O)-C1-C10 alkylene-C(=O)-P1-P2-PAB-; -(C=O)-C1-C10 alkylene-C(=O)NH-P1-P2-PAB-; -(C=O)-C1-C10 alkylene-C(=O)-P1-P2-PAB-DMEDA-; -(C=O)-C1-C10 alkylene-C(=O)NH-P1-P2-PAB-DMEDA-; -(C=O)-C1-C10 alkylene-C(=O)-P1-P2-NMEDA-; -(C=O)-C1-C10 alkylene-C(=O)NH-P1-P2-NMEDA-; m is any integer selected from 3 to 15, preferably 5-10, 6-9, 7-8, n is any integer selected from 1 to 10, preferably 2-6, 2-5 or 2-4; Preferably, the linker is selected from: -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-P1-P2-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-P1-P2-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-P1-P2-PAB-DMEDA-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-P1-P2-NMEDA-; -(C=O)-C6alkylene-C(=O)-P1-P2-; -(C=O)-C6 alkylene-C(=O)-P1-P2-PAB-; -(C=O)-C6 alkylene-C(=O)-P1-P2-PAB-DMEDA-; -(C=O)-C6alkylene-C(=O)-P1-P2-NMEDA-; -(Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-P1-P2-; -(Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-P1-P2-PAB-; -(Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-P1-P2-PAB-DMEDA-; -(Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-P1-P2-NMEDA-; 20. The linker according to any one of claims 9 to 19, wherein The linker is selected from: -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAGLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPLGLGGFG-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPASLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRGLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPSALVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRGLGGFG-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAALGGFG-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRNLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNDap(Me2)VCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRNLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPLSLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRGLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRPLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRSLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRPLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPCitPLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-HPRGLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-HPCitPLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitGLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAPLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PLSLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitSLVCit-PAB-; -(C=O)-C6alkylene-C(=O)-GPRNLVCit-PAB-; -(Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-GPRNLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLVCit-PAB-DMEDA-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLGGFG-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLGGFG-NMEDA-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRSLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRSLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PCitSLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PCitPLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PCitGLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPCitGLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRGLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRPLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitPLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPANLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPCitNLVCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitSDap(Me2)VCit-PAB-; -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRSDap(Me2)VCit-PAB-; -(Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-PCitNLVCit-PAB-; and -(Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PASLVCit-PAB-; Preferably, the linker is selected from: -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLGLGGFG-; -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLGGFG-; -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAALGGFG-; -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-; -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPASLVCit-PAB-; -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLVCit-PAB-; -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPSALVCit-PAB-; -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-PRNLVCit-PAB-; -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRNLVCit-PAB-; -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGDap(Me2)VCit-PAB-; -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-SPRNLVCit-PAB-; -(Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLSLVCit-PAB-.

21. A linker drug comprising a polypeptide or polypeptide fragment having a structure represented by -P1-P2- as claimed in any one of claims 1 to 8, wherein: The linker drug has the following structure: C1'-La-L1-P1-P2-L2-D Wherein, the C1' is a functional group that reacts with a biological ligand; La is empty or a tuning spacer; L1 is empty or a spacer; L2 is empty or a spacer; D is the drug unit; The positions of La and L1 can be swapped.

22. The linker drug according to claim 21, wherein The La is as defined in claim 12, the L1 is as defined in claim 13, and the L2 is as defined in any one of claims 15-17.

23. The linker drug according to claim 21 or 22, wherein C1' is selected from the functional groups that react with amino, thiol, keto, azide and alkyne.

24. The linker drug according to any one of claims 21 to 23, wherein C1' is selected from the following structures: Among them, R X is selected from halogen, -SPh; R S Selected from halogen, sulfone, tertiary amine, diazonium, -OMs, MeSO2-, and CF3SO3-; Indicates the connection position with La; Preferably, C1' is selected from the following structures:

25. The linker drug according to any one of claims 21 to 24, wherein The C1'-La-L1- is MC-, MeO2S-Pym-, NHS-, Mal-PEG8-, DBCO- or Hydrazide-.

26. The linker drug according to any one of claims 21 to 25, wherein The drug unit D is selected from immunomodulators, protein degraders, and cytotoxic agents.

27. The linker drug according to any one of claims 21 to 26, wherein The drug unit D includes: amanitins, anthracyclines, auristatins, baccatins, calicheamicins, camptothecins, cemadotins, colchicine, colchicines, colcimids, combretastatins, cryptophycins, discodermolides, duocarmycins, docetaxel, doxorubicin, duocarmycins, echinomycins, eloserol eleutherobins, epothilones, estramustines, lexitropsins, maytansines, maytansinoids, methotrexate, netropsins, pyrrolo[2,1-c][1,4]benzodi-azepines (PBDs), puromycins, rhizoxins, SN-38, taxanes, tubulysins, vincaalkaloids, tetrahydroisoquinoline alkaloids or derivatives thereof, protein degraders or derivatives thereof.

28. The linker drug according to any one of claims 21 to 27, wherein The drug unit D includes maytansine alkaloids or derivatives thereof, camptothecin or derivatives thereof, auristatin or derivatives thereof, tetrahydroisoquinoline alkaloids or derivatives thereof, BTK protein degraders and derivatives thereof, and GSPT1 protein degraders and derivatives thereof.

29. The linker drug according to any one of claims 21 to 28, wherein The drug unit D includes DM1, DM4, DX8951, MMAE, Dxd, SN38, Trabectedin (ET743), Lurbinectedin, CC885, and CC-90009.

30. The linker drug according to any one of claims 21 to 29, wherein The linker drug is selected from: MC-P1-P2-D; MC-P1-P2-PAB-D; MC-P1-P2-PAB-DMEDA-D; MC-P1-P2-NMEDA-D; MeO2S-Pym-P1-P2-D; MeO2S-Pym-P1-P2-PAB-D; MeO2S-Pym-P1-P2-PAB-DMEDA-D; MeO2S-Pym-P1-P2-NMEDA-D; NHS-P1-P2-D; NHS-P1-P2-PAB-D; NHS-P1-P2-PAB-DMEDA-D; NHS-P1-P2-NMEDA-D; Mal-PEG8-P1-P2-D; Mal-PEG8-P1-P2-PAB-D; Mal-PEG8-P1-P2-PAB-DMEDA-D; Mal-PEG8-P1-P2-NMEDA-D; DBCO-P1-P2-D; DBCO-P1-P2-PAB-D; DBCO-P1-P2-PAB-DMEDA-D; DBCO-P1-P2-NMEDA-D; Hydrazide-P1-P2-D; Hydrazide-P1-P2-PAB-D; Hydrazide-P1-P2-PAB-DMEDA-D; and Hydrazide-P1-P2-NMEDA-D Preferably, the drug unit D comprises DX8951, MMAE, Dxd, SN38 or ET743.

31. The linker drug according to any one of claims 21 to 30, wherein The linker drug has the following structure: (Maleimido-N-yl)-CH2-C(=O)-P1-P2-D; (Maleimido-N-yl)-CH2CH2-C(=O)-P1-P2-D; (Maleimido-N-yl)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D; (Maleimide-N-yl)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D; (Maleimido-N-yl)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D; (Maleimido-N-yl)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D; (Maleimido-N-yl)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D; (Maleimido-N-yl)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D; (Maleimido-N-yl)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-D; (Maleimido-N-yl)-CH2-C(=O)-P1-P2-PAB-D; (Maleimido-N-yl)-CH2CH2-C(=O)-P1-P2-PAB-D; (Maleimide-N-yl)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D; (Maleimide-N-yl)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D; (Maleimido-N-yl)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D; (Maleimido-N-yl)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D; (Maleimido-N-yl)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D; (Maleimide-N-yl)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D; (Maleimido-N-yl)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-PAB-D; (N-hydroxysuccinimide ester-1-yl)-CH2-C(=O)-P1-P2-D; (N-hydroxysuccinimide ester-1-yl)-CH2CH2-C(=O)-P1-P2-D; (N-hydroxysuccinimide ester-1-yl)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D; (N-hydroxysuccinimide ester-1-yl)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D; (N-hydroxysuccinimide ester-1-yl)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D; (N-hydroxysuccinimide ester-1-yl)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D; (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2-C(=O)-P1-P2-D; (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D; (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D; (N-hydroxysuccinimidyl ester-1-yl)-(1,4-cyclohexyl)-CH2C(=O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D; (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2C(=O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D; (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2O-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D; (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2O-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D; (N-hydroxysuccinimide ester-1-yl)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-D; (N-hydroxysuccinimide ester-1-yl)-CH2-C(=O)-P1-P2-PAB-D; (N-hydroxysuccinimide ester-1-yl)-CH2CH2-C(=O)-P1-P2-PAB-D; (N-hydroxysuccinimide ester-1-yl)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D; (N-hydroxysuccinimide ester-1-yl)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D; (N-hydroxysuccinimide ester-1-yl)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D; (N-hydroxysuccinimide ester-1-yl)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D; (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2-C(=O)-P1-P2-PAB-D; (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D; (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D; (N-hydroxysuccinimidyl ester-1-yl)-(1,4-cyclohexyl)-CH2C(=O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D; (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2C(=O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D; (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2O-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D; (N-hydroxysuccinimide ester-1-yl)-(1,4-cyclohexyl)-CH2O-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D; (N-hydroxysuccinimide ester-1-yl)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-PAB-D; Gly-Gly-Gly-NH-CH2CH2CH2CH2CH2-C(=O)-P1-P2-D; Gly-Gly-Gly-NH-CH2CH2CH2CH2CH2-C(=O)-P1-P2-PAB-D; The D is camptothecin or its derivatives, auristatin or its derivatives, tetrahydroisoquinoline alkaloids or their derivatives, protein degraders and their derivatives, preferably DX8951, MMAE, Dxd, SN38, ET743, Lurbinectedin, CC885, CC-90009, more preferably DX8951, MMAE, Dxd, SN38 or ET743.

32. The linker drug according to any one of claims 21 to 31, wherein The linker drug has the following structure: MC-GPAGLVCit-PAB-DX8951; MC-GPAGLVCit-PAB-MMAE; MC-GPAGLVCit-PAB-ET743; MC-GPLGLGGFG-DX8951; MC-GPASLVCit-PAB-DX8951; MC-GPRGLVCit-PAB-DX8951; MC-GPRGLVCit-PAB-MMAE; MC-GPSALVCit-PAB-DX8951; MC-GPRGLGGFG-DX8951; MC-GPAALGGFG-DX8951; MC-PRNLVCit-PAB-DX8951; MC-PRNLVCit-PAB-MMAE; MC-GPRNLVCit-PAB-DX8951; MC-GPRNLVCit-PAB-MMAE; MC-GPRNLVCit-PAB-ET743; MC-GPRNDap(Me2)VCit-PAB-DX8951; MC-GPRNDap(Me2)VCit-PAB-MMAE; MC-SPRNLVCit-PAB-DX8951; MC-SPRNLVCit-PAB-MMAE; MC-GPLSLVCit-PAB-DX8951; MC-GPLSLVCit-PAB-MMAE; MC-PRGLVCit-PAB-DX8951; MC-PRPLVCit-PAB-DX8951; MC-GPRSLVCit-PAB-DX8951; MC-GPRPLVCit-PAB-DX8951; MC-GPCitPLVCit-PAB-DX8951; MC-HPRGLVIt-PAB-DX8951; MC-HPCitPLVCit-PAB-DX8951; MC-SPCitGLVCit-PAB-DX8951; MC-GPAPLVCit-PAB-DX8951; MC-PLSLVCit-PAB-DX8951; MC-SPCitSLVCit-PAB-DX8951; MC-SPCitSLVCit-PAB-MMAE; MC-SPCitSLVCit-PAB-ET743; MeO2S-Pym-GPRNLVCit-PAB-DX8951; DBCO-GPRNLVCit-PAB-DX8951; NHS-GPRNLVCit-PAB-DX8951; Hydrazide-GPRNLVCit-PAB-DX8951; Mal-PEG8-GPRNLVCit-PAB-DX8951; MC-GPRNLVCit-PAB-DMEDA-SN38; MC-GPRNLGGFG-Dxd; MC-GPRNLGGFG-NMEDA-Dxd; MC-PRSLVCit-PAB-DX8951; MC-SPRSLVCit-PAB-DX8951 MC-PCitSLVCit-PAB-DX8951; MC-PCitPLVCit-PAB-DX8951; MC-PCitGLVCit-PAB-DX8951; MC-GPCitGLVCit-PAB-DX8951; MC-SPRGLVCit-PAB-DX8951; MC-SPRPLVCit-PAB-DX8951; MC-SPCitPLVCit-PAB-DX8951; MC-GPANLVCit-PAB-DX8951; MC-GPCitNLVCit-PAB-DX8951; MC-SPCitSDap(Me2)VCit-PAB-DX8951; MC-SPCitSDap(Me2)VCit-PAB-MMAE; MC-PRSDap(Me2)VCit-PAB-DX8951; MC-PRSDap(Me2)VCit-PAB-MMAE; MC-PCitNLVCit-PAB-DX8951; and MC-PASLVCit-PAB-DX8951; Preferably, the linker drug has the following structure: MC-GPAGLVCit-PAB-DX8951; MC-GPLGLGGFG-DX8951; MC-GPASLVCit-PAB-DX8951; MC-GPRGLVCit-PAB-DX8951; MC-GPSALVCit-PAB-DX8951; MC-GPRGLGGFG-DX8951; MC-GPAALGGFG-DX8951; MC-PRNLVCit-PAB-DX8951; MC-GPRNLVCit-PAB-DX8951; MC-GPRGDap(Me2)LVCit-PAB-DX8951; MC-SPRNLVCit-PAB-DX8951; MC-GPLSLVCit-PAB-DX8951; MC-PRGLVCit-PAB-DX8951; MC-PRPLVCit-PAB-DX8951; MC-GPRSLVCit-PAB-DX8951; MC-GPRPLVCit-PAB-DX8951; MC-GPCitPLVCit-PAB-DX8951; MC-HPRGLVIt-PAB-DX8951; MC-HPCitPLVCit-PAB-DX8951; MC-SPCitGLVCit-PAB-DX8951; MC-GPAPLVCit-PAB-DX8951; MC-PLSLVCit-PAB-DX8951; MC-SPCitSLVCit-PAB-DX8951; MC-PRSLVCit-PAB-DX8951; MC-SPRSLVCit-PAB-DX8951; MC-PCitSLVCit-PAB-DX8951; MC-PCitPLVCit-PAB-DX8951; MC-PCitGLVCit-PAB-DX8951; MC-GPCitGLVCit-PAB-DX8951; MC-SPRGLVCit-PAB-DX8951; MC-SPRPLVCit-PAB-DX8951; MC-SPCitPLVCit-PAB-DX8951; MC-GPANLVCit-PAB-DX8951; MC-GPCitNLVCit-PAB-DX8951; MC-SPCitSDap(Me2)VCit-PAB-DX8951; MC-PRSDap(Me2)VCit-PAB-DX8951; MC-GPAGLVCit-PAB-MMAE; MC-GPRGLVCit-PAB-MMAE; MC-PRNLVCit-PAB-MMAE; MC-GPRNLVCit-PAB-MMAE; MC-GPRGDap(Me2)LVCit-PAB-MMAE; MC-SPRNLVCit-PAB-MMAE; MC-GPLSLVCit-PAB-MMAE; MC-SPCitSLVCit-PAB-MMAE MC-SPCitSDap(Me2)VCit-PAB-MMAE MC-SPCitSDap(Me2)VCit-PAB-MMAE MC-GPAGLVCit-PAB-ET743; MC-GPRNLVCit-PAB-ET743; MC-SPCitSLVCit-PAB-ET743.

33. A bioligand-drug conjugate comprising the linker of any one of claims 9 to 20, wherein: The conjugate has the following general formula: in: Bp is the biological ligand; L is the linker; D is the drug unit; p is any integer between 1 and 20.

34. The conjugate according to claim 33, wherein the conjugate has the general formula: in: Bp is the biological ligand; C1-La-L1-P1-P2-L2 is the linker; D is the drug unit; p is any integer between 1 and 20.

35. The conjugate according to claim 33 or 34, wherein The drug unit D is as defined in any one of claims 26-31.

36. The conjugate according to any one of claims 33 to 35, wherein The biological ligand targets cell surface receptors or tumor-associated antigens, wherein the cell surface receptors or tumor-associated antigens include: EGFR, HER2, HER3, c-Met, claudin18.2, TROP-2, folate receptor α (FRα), ROR1, ROR2, BCMA, PSMA, CD19, CD20, CD22, CD30, CD33, CD37, CD46, CD48, CD56, CD79b, CD123, CD138, CD166, CD276, CEACAM5, SDC1, CD74, CD70, MUC1, MUC16, GUCY2C, MSLN, SCL34A2, FOLH1, TPBC, PVRL4, STEAP1, SCL44A4, NACM1, EDNRB, GPNMB, FGFR, SEZ6, Nectin-4, PD-L1, Tissue Factor(TF), B7-H3, B7-H4, LY6E, LIV-1, CDH4, CDH6, ITGB6, GPC1, GPC3, ENPP3, KAAG1, FZD7, SSTR2, DLK1, T MEFF1, TM4SF1, SLAMF7, DLL3, FLT3, TNFRSF10B, EPCAM, AXL, IL2RA, 5T4, FAP, ICAM-1, IGF-1R, TNFR1, GLP-1.

37. The conjugate according to any one of claims 33 to 36, wherein The biological ligands include antibodies or antigen-binding fragments thereof, modified antibodies or antigen-binding fragments thereof, oligonucleotides, DNA fragments, nucleic acid aptamers, polypeptides, nanoparticles with targeting properties, and chemically synthesized small molecules.

38. The conjugate according to claim 37, wherein The antigen binding fragments include Fab, Fab', Fv fragment, F(ab')2, scFv, di-scFv and / or dAb.

39. The conjugate according to claim 37 or 38, wherein The antibody or antigen-binding fragment thereof, modified antibody or modified antigen-binding fragment thereof is: anti-EGFR, anti-HER2, anti-HER3, anti-c-Met, anti-claudin18.2, anti-TROP-2, anti-FRα, anti-ROR1, anti-ROR2, anti-BCMA, anti-PSMA, anti-CD19, anti-CD20, anti-CD22, anti- CD30, anti-CD33, anti-CD37, anti-CD46, anti-CD48, anti-CD56, anti-CD79b, anti-CD123, anti-CD138, anti-CD166, anti-CD276, anti-CEACAM5, anti-SDC1, anti-CD74, anti-CD70, anti-MUC1, anti-M UC16, anti-GUCY2C, anti-MSLN, anti-SCL34A2, anti-FOLH1, anti-TPBC, anti-PVRL4, anti-STEAP1, anti-SCL44A4, anti-NACM1, anti-EDNRB, anti-GPNMB, anti-FGFR, anti-SEZ6, anti-Nectin-4, anti- PD-L1, anti-TF, anti-B7-H3, anti-B7-H4, anti-LY6E, anti-LIV-1, anti-CDH4, anti-CDH6, anti-ITGB6, anti-GPC1, anti-GPC3, anti-ENPP3, anti-KAAG1, anti-FZD7, anti-SSTR2, anti-DLK1, anti-TME Antibodies or antigen-binding fragments of FF1, anti-TM4SF1, anti-SLAMF7, anti-DLL3, anti-FLT3, anti-TNFRSF10B, anti-EPCAM, anti-AXL, anti-IL2RA, anti-5T4, anti-FAP, anti-ICAM-1, anti-IGF-1R, anti-TNFR1, anti-GLP-1.

40. The conjugate according to any one of claims 37 to 39, wherein The antibody or antigen-binding fragment thereof, modified antibody or modified antigen-binding fragment thereof is: anti-Her2, anti-TF, anti-ROR1, anti-GPC3 antibody or antigen-binding fragment thereof.

41. The conjugate according to any one of claims 37 to 40, wherein The antibody is Trastuzumab, Tisotumab, Zilovertamab or Codrituzumab.

42. The conjugate according to any one of claims 33 to 41, wherein The conjugate is an antibody-drug conjugate, and the antibody-drug conjugate is selected from: as well as Preferably, the antibody drug conjugate is selected from the following structures: Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-P1-P2-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-P1-P2-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-P1-P2-PAB-DMEDA-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-P1-P2-NMEDA-D) n ; Ab-((C=O)-C6 alkylene-C(=O)-D) n ; Ab-((C=O)-C6 alkylene-C(=O)-P1-P2-PAB-D) n ; Ab-((C=O)-C6 alkylene-C(=O)-P1-P2-PAB-DMEDA-D) n ; Ab-((C=O)-C6alkylene-C(=O)-P1-P2-NMEDA-D) n ; Ab-((Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-P1-P2-D) n ; Ab-((Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-P1-P2-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-P1-P2-PAB-DMEDA-D) n ; Ab-((Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-P1-P2-NMEDA-D) n ; The Ab represents a monoclonal antibody or an antigenic structure fragment thereof, or a modified monoclonal antibody or an antigen-binding fragment thereof, preferably an anti-Her2, anti-TF, anti-ROR1 or anti-GPC3 antibody or an antigenic structure fragment thereof; the D is camptothecin or a derivative thereof, auristatin or a derivative thereof, a tetrahydroisoquinoline alkaloid or a derivative thereof, a protein degrader and a derivative thereof, preferably DX8951, MMAE, Dxd, SN38, ET743, Lurbinectedin, CC885, CC-90009; the n represents any value selected from 1-10.

43. The conjugate according to any one of claims 33 to 42, wherein The conjugate is an antibody-drug conjugate, and the antibody-drug conjugate is selected from the following structures: Ab-((Succinimidyl-3-yl-N)-CH2-C(=O)-P1-P2-D) n ; Ab-((Succinimidyl-3-yl-N)-CH2CH2-C(=O)-P1-P2-D) n ; Ab-((Succinimidyl-3-yl-N)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D) n ; Ab-((Succinimidyl-3-yl-N)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D) n ; Ab-((Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D) n ; Ab-((Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D) n ; Ab-((Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-D) n ; Ab-((Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-D) n ; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-D) n ; Ab-((Succinimidyl-3-yl-N)-CH2-C(=O)-P1-P2-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-CH2CH2-C(=O)-P1-P2-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-CH2CH2C(O)NH-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)4-CH2CH2-C(=O)-P1-P2-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-OC(O)-(CH2CH2O)8-CH2CH2-C(=O)-P1-P2-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-P1-P2-PAB-D) n ; The Ab represents a monoclonal antibody or an antigenic structural fragment thereof, or a modified monoclonal antibody or an antigen-binding fragment thereof, preferably an anti-Her2, anti-TF, anti-ROR1, anti-GPC3 antibody or an antigen-binding fragment thereof; The -P1-P2- represents the following structure: -His-Val-Leu-Asn-Leu-Val-Cit-(SEQ ID NO: 1); -Val-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:2); -Ile-Pro-Val-Ser-Leu-Val-Cit-(SEQ ID NO:3); -Gly-Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:4); -Gly-Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:5); -Gly-Pro-Ala-Gly-Leu-Val-Cit-(SEQ ID NO: 6); -Gly-Pro-Gln-Gly-Leu-Val-Cit-(SEQ ID NO:7); -Gly-Pro-Leu-Gly-Leu-Val-Cit-(SEQ ID NO:8); -Gly-Pro-Ser-Gly-Leu-Val-Cit-(SEQ ID NO:9); -Gly-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO: 10); -Gly-Pro-Ala-Ala-Leu-Val-Cit-(SEQ ID NO: 11); -Gly-Pro-Ser-Ala-Leu-Val-Cit-(SEQ ID NO:12); -His-Val-Leu-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO: 13); -Val-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO: 14); -Ile-Pro-Val-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:15); -Gly-Pro-Leu-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:16); -Gly-Pro-Ala-Ser-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:17); -Gly-Pro-Ala-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:18); -Gly-Pro-Gln-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:19); -Gly-Pro-Leu-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:20); -Gly-Pro-Ser-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:21); -Gly-Pro-Arg-Gly-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:22); -Gly-Pro-Ala-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:23); -Gly-Pro-Ser-Ala-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:24); -Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:25); -Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:26); -Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:27); -Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:28); -Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:29); -Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:30); -Gly-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:31); -Gly-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:32); -Gly-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:33); -Gly-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:34); -Gly-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:35); -His-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:36); -His-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:37); -His-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:38); -His-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:39); -His-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:40); -His-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:41); -Ser-Pro-Arg-Gly-Leu-Val-Cit-(SEQ ID NO:42); -Ser-Pro-Arg-Ser-Leu-Val-Cit-(SEQ ID NO:43); -Ser-Pro-Arg-Pro-Leu-Val-Cit-(SEQ ID NO:44); -Ser-Pro-Arg-Asn-Leu-Val-Cit-(SEQ ID NO:45); -Ser-Pro-Cit-Gly-Leu-Val-Cit-(SEQ ID NO:46); -Ser-Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:47); -Ser-Pro-Cit-Pro-Leu-Val-Cit-(SEQ ID NO:48); -Gly-Pro-Ala-Pro-Leu-Val-Cit-(SEQ ID NO:49); -Gly-Pro-Ala-Asn-Leu-Val-Cit-(SEQ ID NO:50); -Gly-Pro-Arg-Gly-Dap(Me2)-Val-Cit-(SEQ ID NO:51); -Gly-Pro-Arg-Asn-Dap(Me2)-Val-Cit-(SEQ ID NO:60); -Pro-Leu-Ser-Leu-Val-Cit-(SEQ ID NO:61); -Gly-Pro-Arg-Asn-Leu-Gly-Gly-Phe-Gly-(SEQ ID NO:62); -Pro-Cit-Ser-Leu-Val-Cit-(SEQ ID NO:63); -Gly-Pro-Cit-Asn-Leu-Val-Cit-(SEQ ID NO:64); -Ser-Pro-Cit-Ser-Dap(Me2)-Val-Cit-(SEQ ID NO:65); -Pro-Arg-Ser-Dap(Me2)-Val-Cit-(SEQ ID NO:66); -Pro-Cit-Asn-Leu-Val-Cit-(SEQ ID NO:67); -Pro-Ala-Ser-Leu-Val-Cit-(SEQ ID NO:68); The D is camptothecin or its derivatives, auristatin or its derivatives, tetrahydroisoquinoline alkaloids or their derivatives, protein degraders and their derivatives, preferably DX8951, MMAE, Dxd, SN38, ET743, Lurbinectedin, CC885, CC-90009; Said n represents an integer of 1-10.

44. The conjugate according to any one of claims 33 to 43, wherein The conjugate is an antibody-drug conjugate, and the antibody-drug conjugate is selected from the following structures: Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAGLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPLGLGGFG-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPASLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRGLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPSALVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRGLGGFG-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAALGGFG-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRNLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNDap(Me2)VCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRNLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPLSLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRGLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRPLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRSLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRPLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPCitPLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-HPRGLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-HPCitPLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitGLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAPLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PLSLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitSLVCit-PAB-D) n ; Ab-((C=O)-C6alkylene-C(=O)-GPRNLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-GPRNLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLVCit-PAB-DMEDA-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLGGFG-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLGGFG-NMEDA-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRSLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRSLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PCitSLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PCitPLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PCitGLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPCitGLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRGLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRPLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitPLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPANLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPCitNLVCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitSDap(Me2)VCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRSDap(Me2)VCit-PAB-D) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PCitNLVCit-PAB-D) n ;as well as Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PASLVCit-PAB-D) n ; The Ab is an anti-Her2, anti-TF, anti-ROR1 or anti-GPC3 antibody or an antigenic structural fragment thereof; the D is DX8951, MMAE, Dxd, SN38 or ET743; the n represents any value selected from 2-9, preferably 2-8; Preferably, the antibody drug conjugate is selected from the following structures: Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-GPAGLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAGLVCit-PAB-MMAE) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAGLVCit-PAB-ET743) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-GPLGLGGFG-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-GPASLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-GPRGLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRGLVCit-PAB-MMAE) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPSALVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-GPRGLGGFG-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-GPAALGGFG-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRNLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRNLVCit-PAB-MMAE) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-GPRNLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLVCit-PAB-MMAE) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLVCit-PAB-ET743) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-GPRNDap(Me2)VCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNDap(Me2)VCit-PAB-MMAE) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-SPRNLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRNLVCit-PAB-MMAE) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPLSLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPLSLVCit-PAB-MMAE) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRGLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRPLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-GPRSLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRPLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-GPCitPLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-HPRGLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-HPCitPLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-SPCitGLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPAPLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-PLSLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitSLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitSLVCit-PAB-MMAE) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitSLVCit-PAB-ET743) n ; Ab-((C=O)-C6 alkylene-C(=O)-GPRNLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C2alkylene-C(=O)NH-(CH2CH2O)8-C2alkylene-C(=O)-GPRNLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-GPRNLVCit-PAB-DMEDA-SN38) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLGGFG-Dxd) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-GPRNLGGFG-NMEDA-Dxd) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRSLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-SPRSLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-PCitSLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-PCitPLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-PCitGLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-GPCitGLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRGLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPRPLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitPLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-GPANLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-GPCitNLVCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitSDap(Me2)VCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-SPCitSDap(Me2)VCit-PAB-MMAE) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRSDap(Me2)VCit-PAB-DX8951) n ; Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PRSDap(Me2)VCit-PAB-MMAE) n ; Ab-((Succinimidyl-3-yl-N)-C5 alkylene-C(=O)-PCitNLVCit-PAB-DX8951) n ;as well as Ab-((Succinimidyl-3-yl-N)-C5alkylene-C(=O)-PASLVCit-PAB-DX8951) n ; The Ab is an anti-Her2, anti-TF, anti-ROR1 or anti-GPC3 antibody or an antigenic structural fragment thereof; and n represents any value selected from 2-9.

45. The conjugate according to any one of claims 33 to 44, wherein The conjugate is an antibody-drug conjugate, and the antibody-drug conjugate is selected from the following structures: Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-DX8951)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLGLGGFG-DX8951)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPASLVCit-PAB-DX8951)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLVCit-PAB-DX8951)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPSALVCit-PAB-DX8951)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLGGFG-DX8951)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAALGGFG-DX8951)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-PRNLVCit-PAB-DX8951)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRNLVCit-PAB-DX8951)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGDap(Me2)LVCit-PAB-DX8951)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-SPRNLVCit-PAB-DX8951)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLSLVCit-PAB-DX8951)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-MMAE)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGLVCit-PAB-MMAE)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-PRNLVCit-PAB-MMAE)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRNLVCit-PAB-MMAE)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRGDap(Me2)LVCit-PAB-MMAE)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-SPRNLVCit-PAB-MMAE)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPLSLVCit-PAB-MMAE)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPAGLVCit-PAB-ET743)n; Ab-((Succinimidyl-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GPRNLVCit-PAB-ET743)n; The Ab represents a monoclonal antibody or an antigenic structural fragment thereof, or a modified monoclonal antibody or an antigen-binding fragment thereof, preferably an anti-Her2, anti-TF, anti-ROR1, anti-GPC3 antibody or an antigen-binding fragment thereof; Said n is an integer of 1-10; preferably, n is 2, 4, 6, or 8.

46. ​​An anti-tissue factor (TF) antibody or an antigen-binding fragment thereof, comprising: (a) VH comprising the amino acid sequence shown in SEQ ID NO: 52 or 54; and / or (b) VL comprising the amino acid sequence shown in SEQ ID NO: 53 or 55.

47. The anti-TF antibody or antigen-binding fragment thereof according to claim 46, wherein: The anti-TF antibody or antigen-binding fragment thereof comprises: (1) VH and VL, wherein the VH has the amino acid sequence shown in SEQ ID NO: 52, and the VL has the amino acid sequence shown in SEQ ID NO: 53; or (2) VH and VL, wherein the VH has the amino acid sequence shown in SEQ ID NO: 54, and the VL has the amino acid sequence shown in SEQ ID NO: 53; or (3) VH and VL, wherein the VH has the amino acid sequence shown in SEQ ID NO: 54, and the VL has the amino acid sequence shown in SEQ ID NO: 55; or (4) VH and VL, wherein the VH has the amino acid sequence shown in SEQ ID NO: 52, and the VL has the amino acid sequence shown in SEQ ID NO:

55.

48. The anti-TF antibody or antigen-binding fragment thereof according to claim 46 or 47, wherein: The anti-TF antibody or antigen-binding fragment thereof comprises a light chain as shown in SEQ ID NO: 56, or an amino acid sequence having at least 80% identity with the amino acid sequence as shown in SEQ ID NO: 56; and a heavy chain as shown in SEQ ID NO: 57, or an amino acid sequence having at least 80% identity with the amino acid sequence as shown in SEQ ID NO:

57.

49. The anti-TF antibody or antigen-binding fragment thereof according to claim 46 or 47, wherein: The anti-TF antibody or antigen-binding fragment thereof comprises a light chain as shown in SEQ ID NO: 56, or an amino acid sequence having at least 80% identity with the amino acid sequence as shown in SEQ ID NO: 56; and a heavy chain as shown in SEQ ID NO: 59, or an amino acid sequence having at least 80% identity with the amino acid sequence as shown in SEQ ID NO:

59.

50. The anti-TF antibody or antigen-binding fragment thereof according to claim 46 or 47, wherein: The anti-TF antibody or antigen-binding fragment thereof comprises a light chain as shown in SEQ ID NO: 58, or an amino acid sequence having at least 80% identity with the amino acid sequence as shown in SEQ ID NO: 58; and a heavy chain as shown in SEQ ID NO: 59, or an amino acid sequence having at least 80% identity with the amino acid sequence as shown in SEQ ID NO:

59.

51. A nucleic acid molecule encoding the anti-TF antibody or antigen-binding fragment thereof according to any one of claims 46 to 50, or the heavy chain variable region and / or light chain variable region thereof.

52. A recombinant vector comprising the nucleic acid molecule of claim 51.

53. A recombinant cell comprising the nucleic acid molecule of claim 51 or the recombinant vector of claim 52.

54. A method for preparing an anti-TF antibody or an antigen-binding fragment thereof, the method comprising culturing the recombinant cell of claim 53.

55. Use of the anti-TF antibody or antigen-binding fragment thereof according to any one of claims 46 to 50, the nucleic acid molecule according to claim 51, the recombinant vector according to claim 52 or the recombinant cell according to claim 53 in the preparation of a medicament; preferably, the medicament is a medicament for preventing and / or treating cancer; more preferably, the cancer is a cancer expressing TF.

56. Use of the anti-TF antibody or antigen-binding fragment thereof according to any one of claims 46 to 50, the nucleic acid molecule according to claim 51, the recombinant vector according to claim 52 or the recombinant cell according to claim 53 in the preparation of a reagent for diagnosing a cell proliferation-related disease; preferably, the disease is cancer; more preferably, the cancer is a cancer expressing TF.

57. A pharmaceutical composition comprising the polypeptide or polypeptide fragment of any one of claims 1 to 8, the linker of any one of claims 9 to 20, the linker drug of any one of claims 21 to 32, the conjugate of any one of claims 33 to 45, the anti-TF antibody or antigen-binding fragment thereof of any one of claims 46 to 50, the nucleic acid molecule of claim 51, the recombinant vector of claim 52, the recombinant cell of claim 53, and optionally one or more pharmaceutical excipients; preferably, it also comprises a pharmaceutically acceptable carrier and / or excipient.

58. Use of the polypeptide or polypeptide fragment according to any one of claims 1 to 8, the linker according to any one of claims 9 to 20, the linker drug according to any one of claims 21 to 32, the conjugate according to any one of claims 33 to 45, the anti-TF antibody or antigen-binding fragment thereof according to any one of claims 46 to 50, the nucleic acid molecule according to claim 51, the recombinant vector according to claim 52, the recombinant cell according to claim 53, or the pharmaceutical composition according to claim 57 in the preparation of a drug for treating / preventing a disease or condition.

59. Use of the polypeptide or polypeptide fragment according to any one of claims 1 to 8, the linker according to any one of claims 9 to 20, the linker drug according to any one of claims 21 to 32, the conjugate according to any one of claims 33 to 45, the anti-TF antibody or antigen-binding fragment thereof according to any one of claims 46 to 50, the nucleic acid molecule according to claim 51, the recombinant vector according to claim 52, the recombinant cell according to claim 53, or the pharmaceutical composition according to claim 57 in the preparation of a medicament for treating / preventing a disease or condition associated with abnormal cell activity.

60. The use according to claim 58 or 59, wherein The disease or condition includes a tumor.

61. The use according to claim 60, wherein Such diseases or conditions include solid tumors and hematological tumors.

62. A method for treating a disease or disorder, preferably a tumor, in a subject in need thereof, comprising administering to the subject an effective amount of the conjugate of any one of claims 33-45.

63. The use according to any one of claims 58 to 61 or the method according to claim 62, wherein The disease or condition is selected from the following tumors associated with protein expression: EGFR, HER2, HER3, c-Met, claudin18.2, TROP-2, folate receptor α (FRα), ROR1, ROR2, BCMA, PSMA, CD19, CD20, CD22, CD30, CD33, CD37, CD46, CD48, CD56, CD79b, CD123, CD138, CD166, CD276, CEACAM5, SDC1, CD74, CD70, MUC1, MUC16, GUCY2C, MSLN, SCL34A2, FOLH1, TPBC, PVRL4, STEAP1, SCL44A4, NACM1, EDNRB, GPNMB, FGFR, SEZ6, Nectin-4, PD-L1, Tissue Factor(TF), B7-H3, B7-H4, LY6E, LIV-1, CDH4, CDH6, ITGB6, GPC1, GPC3, ENPP3, KAAG1, FZD7, SSTR2, DLK1, TMEFF1, TM4SF1, SLAMF7, DLL3, FLT3, TNFRSF10B, EPCAM, AXL, IL2RA, 5T4, FAP, ICAM-1, IGF-1R; Preferably, the tumor is selected from tumors associated with Her2, TF, ROR1 or GPC3 expression.

64. The use according to any one of claims 58 to 61 or the method according to claim 62, wherein The disease or condition is selected from the group consisting of esophageal cancer, brain cancer, lung cancer, epithelial cell cancer, bladder cancer, gastric cancer, ovarian cancer, pancreatic cancer, head and neck cancer, urothelial cancer, colon cancer, colorectal cancer, cervical cancer, endometrial cancer, rectal cancer, kidney cancer, prostate cancer, melanoma, liver cancer, gallbladder cancer, bile duct cancer, thyroid cancer, non-Hodgkin's lymphoma, multiple myeloma, B-cell lymphoma; Preferably, the tumor is selected from esophageal cancer, lung cancer, breast cancer, gastric cancer, urothelial carcinoma, bile duct cancer, prostate cancer, colorectal cancer, cervical cancer, and ovarian cancer.

65. Use of the polypeptide or polypeptide fragment of any one of claims 1 to 8, the linker of any one of claims 9 to 20, or the linker drug of any one of claims 21 to 32 in the preparation of an antibody-drug conjugate for treating tumors.

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