Antibody-drug conjugates and methods for preparing and using same

Abstract

The present application relates to antibody-drug conjugates and methods for their preparation and use, particularly to antibody-drug conjugates for treating PTK7-positive cancers, comprising a PTK7 antibody and a drug-linker molecule attached to the antibody. In some embodiments, the antibody is humanized and has excellent binding activity to PTK7-positive cells, allowing for efficient drug delivery to PTK7-positive cells. In some embodiments, the drug comprises a DNA topoisomerase inhibitor. The antibody-drug conjugates of the present application have a better drug-antibody binding ratio and exhibit excellent target killing effects against lung cancer, breast cancer, epidermal cancer, ovarian cancer, esophageal cancer, and the like. Therefore, the present application further provides methods for preparing antibody-drug conjugates and their use in treating PTK7-positive cancers.

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims the benefit of Chinese Patent Application No. 202311284321.4, filed on September 28, 2023, and Chinese Patent Application No. 202211263034.0, filed on October 14, 2022, the disclosures of each of which are incorporated herein by reference in their entireties.

[0002] Sequence Listing This application contains a computer-readable sequence listing submitted herewith in XML file format, the entire contents of which are incorporated herein by reference in their entirety. The sequence listing XML file submitted herewith is entitled "14463-051-228_SEQ_LISTING.xml", was created on September 14, 2023, and is 63,238 bytes in size. [Background technology]

[0003] (1) Technical field The present application relates to the field of targeted therapy, specifically to antibody-drug conjugates for treating PTK7-positive cancers. Specifically, the antibody-drug conjugates comprise a PTK7 antibody and a drug-linker molecule attached to the antibody. In some embodiments, the antibody is humanized and has excellent binding activity to PTK7-positive cells, allowing for efficient drug delivery to PTK7-positive cells. In some embodiments, the drug comprises a DNA topoisomerase inhibitor. The antibody-drug conjugates of the present application have an excellent drug-antibody binding ratio and exhibit good target killing effects against lung cancer, breast cancer, epidermal cancer, ovarian cancer, and esophageal cancer. Therefore, the present application further provides methods for preparing the antibody-drug conjugates and their use in treating PTK7-positive cancers.

[0004] (2) Description of related technology Cancer is one of the leading causes of death in modern times. Cancer is a type of disease caused by the malignant transformation of healthy cells. It is caused by genetic alterations such as chromosomal translocations, tumor suppressor genes, and mutations in growth factor receptors, leading to malignant cell proliferation. Incomplete apoptosis, or programmed cell death, further promotes the malignant transformation of cells, resulting in cancer. According to the latest assessment by the International Agency for Research on Cancer (IARC) of the World Health Organization (WHO), there were 19,290,000 new cancer cases worldwide in 2020, including 10,060,000 men and 9,230,000 women. In 2020, 9,960,000 cancer deaths occurred worldwide, including 5,530,000 men and 4,430,000 women. This century, cancer is predicted to surpass cardiovascular disease as the leading cause of premature death in most countries.

[0005] PTK7 (protein tyrosine kinase 7) belongs to the receptor tyrosine kinase family and lacks kinase activity due to mutations in the kinase domain. The PTK7 protein consists of seven extracellular immunoglobulin domains, a transmembrane domain, and an intracellular tyrosine kinase domain, and its ligand is unknown. The extracellular segment of PTK7 can be cleaved by ADAM and MT1-MMP proteases to generate soluble fragments. The serum concentration of free PTK7 in healthy individuals is approximately 12.4±3.3ng / mL, while the concentration in tumor patients is higher, approximately 24.6±3.8ng / mL.

[0006] PTK7 is expressed in various solid tumors. It has been reported to be highly expressed in 47.4% of NSCLCs, 45.1% of ovarian cancers, 28.6% of TNBCs, and 60% of esophageal cancers. Studies have shown that PTK7 expression promotes tumor cell proliferation, and knockout of the PTK7 gene reduces tumor burden in mice. High PTK7 expression positively correlates with disease staging and lymph node metastasis, and negatively correlates with survival. Currently, research is underway on PTK7-targeting antibody-drug conjugates, CAR-T, and other therapeutic approaches. In preclinical studies, tumor cell lines with high PTK7 expression were efficiently killed. It has demonstrated strong tumor-suppressing activity in human-derived tumor cell-derived xenograft (CDX) and human-derived tumor xenograft (PDX) models. Cofetuzumab peridotin, an ADC drug targeting PTK7 developed by Pfizer, has demonstrated good safety and preliminary efficacy in stage I clinical trials.

[0007] ADC drugs are composed of an antibody, a biologically active molecule, and a linker. The biologically active molecule is covalently bound to the antibody via the linker. The antibody (e.g., a monoclonal antibody) can specifically recognize a specific target on the surface of tumor cells, then guide the ADC to the surface of cancer cells, allowing the ADC to enter the cancer cells by endocytosis. The biologically active molecule is then released in the cancer cells, killing the cancer cells with minimal damage to normal tissue cells. Developing antibodies with better targeting, hydrophilicity, and endocytosis activity to bind cytotoxic small molecules to prepare ADC drugs with a wider therapeutic window will provide patients with better treatment options. Summary of the Invention

[0008] This application relates to antibody-drug conjugates for treating PTK7-positive cancers, using humanized antibodies 101A6HZ, 101A6HZm, and 64A10HZ as targeting moieties, and having the general formula Ab-[MLED] xThe present application exemplarily discloses an antibody-drug conjugate having a structure represented by the formula: Figure imgb0001. As shown in the results, the conjugate has a better drug-antibody binding ratio, excellent binding activity to PTK7-positive cells, and good target killing effect on PTK7-positive cells, such as lung cancer, breast cancer, epidermal cancer, ovarian cancer, and esophageal cancer cells. Therefore, the present application provides an antibody-drug conjugate for the treatment of cancers associated with high PTK7 expression, a pharmaceutical composition comprising the antibody-drug conjugate, and its application in the treatment of cancers associated with high PTK7 expression.

[0009] antibody-drug conjugates In one aspect, the present application provides a compound of formula Ab-[MLED] x and providing an antibody-drug conjugate having the structure shown in Ab is an antibody or antigen-binding fragment thereof that specifically binds to human tyrosine kinase 7 (PTK7); M is a linker that is linked to the antibody or antigen-binding fragment thereof; L is a linker between M and E, E is a structural fragment connecting L and D, D is a cytotoxic drug fragment; An antibody-drug conjugate wherein x is selected from 1 to 10.

[0010] In some embodiments, the antibody or antigen-binding fragment thereof (1) A heavy chain variable region (VH) and / or light chain variable region (VL) of the following, wherein the CDRs are defined according to the Chothia numbering system: (1a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence shown as SEQ ID NO: 11 or a variant thereof, CDR-H2 having the sequence shown as SEQ ID NO: 12 or a variant thereof, and CDR-H3 having the sequence shown as SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence shown as SEQ ID NO: 14 or a variant thereof, CDR-L2 having the sequence shown as SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the sequence shown as SEQ ID NO: 16 or a variant thereof; or (1b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence shown as SEQ ID NO: 27 or a variant thereof, CDR-H2 having the sequence shown as SEQ ID NO: 28 or a variant thereof, and CDR-H3 having the sequence shown as SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence shown as SEQ ID NO: 30 or a variant thereof, CDR-L2 having the sequence shown as SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the sequence shown as SEQ ID NO: 32 or a variant thereof; The variants of either (1a) and (1b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with respect to the sequence from which the variant is derived, or have one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, preferably conservative substitutions, in a heavy chain variable region (VH) and / or a light chain variable region (VL); or (2) A heavy chain variable region (VH) and / or a light chain variable region (VL) of the following, wherein the CDRs are defined according to the Kabat numbering system: (2a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence shown as SEQ ID NO: 17 or a variant thereof, CDR-H2 having the sequence shown as SEQ ID NO: 18 or 19 or a variant thereof, and CDR-H3 having the sequence shown as SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence shown as SEQ ID NO: 14 or a variant thereof, CDR-L2 having the sequence shown as SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the sequence shown as SEQ ID NO: 16 or a variant thereof; or (2b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence shown as SEQ ID NO: 33 or a variant thereof, CDR-H2 having the sequence shown as SEQ ID NO: 34 or 35 or a variant thereof, and CDR-H3 having the sequence shown as SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence shown as SEQ ID NO: 30 or a variant thereof, CDR-L2 having the sequence shown as SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the sequence shown as SEQ ID NO: 32 or a variant thereof; The variants of either (2a) and (2b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with respect to the sequence from which the variant is derived, or have one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, preferably conservative substitutions, in a heavy chain variable region (VH) and / or a light chain variable region (VL); or (3) A heavy chain variable region (VH) and / or light chain variable region (VL) of the following, wherein the CDRs are defined according to the IMGT numbering system: (3a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence shown as SEQ ID NO: 20 or a variant thereof, CDR-H2 having the sequence shown as SEQ ID NO: 21 or a variant thereof, and CDR-H3 having the sequence shown as SEQ ID NO: 22 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence shown as SEQ ID NO: 23 or a variant thereof, CDR-L2 having the sequence shown as SEQ ID NO: 24 or a variant thereof, and CDR-L3 having the sequence shown as SEQ ID NO: 16 or a variant thereof; or (3b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence shown as SEQ ID NO: 36 or a variant thereof, CDR-H2 having the sequence shown as SEQ ID NO: 37 or a variant thereof, and CDR-H3 having the sequence shown as SEQ ID NO: 38 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence shown as SEQ ID NO: 39 or a variant thereof, CDR-L2 having the sequence shown as SEQ ID NO: 40 or a variant thereof, and CDR-L3 having the sequence shown as SEQ ID NO: 32 or a variant thereof; The variants of either (3a) and (3b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with respect to the sequence from which the variant is derived, or have one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, and the substitutions are preferably conservative substitutions. or (4) A heavy chain variable region (VH) and / or light chain variable region (VL) of the following, wherein the CDRs are defined according to the AbM numbering system: (4a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence shown as SEQ ID NO: 25 or a variant thereof, CDR-H2 having the sequence shown as SEQ ID NO: 26 or a variant thereof, and CDR-H3 having the sequence shown as SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence shown as SEQ ID NO: 14 or a variant thereof, CDR-L2 having the sequence shown as SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the sequence shown as SEQ ID NO: 16 or a variant thereof; or (4b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence shown as SEQ ID NO: 41 or a variant thereof, CDR-H2 having the sequence shown as SEQ ID NO: 42 or a variant thereof, and CDR-H3 having the sequence shown as SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence shown as SEQ ID NO: 30 or a variant thereof, CDR-L2 having the sequence shown as SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the sequence shown as SEQ ID NO: 32 or a variant thereof; The variants of either (4a) or (4b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence from which the variant is derived, or have undergone one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, and preferably, the substitutions are conservative substitutions. In some embodiments, the antibody or antigen-binding fragment thereof (a) a VH as set forth in SEQ ID NO: 1 or a variant thereof, and / or a VL as set forth in SEQ ID NO: 2 or a variant thereof; (b) a VH as set forth in SEQ ID NO: 3 or a variant thereof, and / or a VL as set forth in SEQ ID NO: 4 or a variant thereof; or (c) VH shown as SEQ ID NO: 5 or a variant thereof, and / or VL shown as SEQ ID NO: 6 or a variant thereof; (d) VH as shown in SEQ ID NO: 7 or a variant thereof, and / or VL as shown in SEQ ID NO: 8 or a variant thereof; (e) comprising a VH set forth as SEQ ID NO: 9 or a variant thereof, and / or a VL set forth as SEQ ID NO: 10 or a variant thereof; A variant may have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence from which it is derived, or may have one or several amino acid substitutions, deletions, or additions (e.g., 1, 2, 3, 4, or 5 amino acid substitutions, deletions, or additions) with respect to the sequence from which it is derived, preferably wherein the substitutions are conservative substitutions.

[0011] In some embodiments, the antibody or antigen-binding fragment thereof (a) a human immunoglobulin heavy chain constant region (CH) or a variant thereof, wherein the variant has one or more amino acid substitutions, deletions, or additions (e.g., up to 20, up to 15, up to 10, or up to 5 amino acid substitutions, deletions, or additions, e.g., 1, 2, 3, 4, or 5 amino acid substitutions, deletions, or additions) compared to the wild-type sequence from which it is derived; and (b) A human immunoglobulin light chain constant region (CL) or variant thereof, wherein the variant further comprises one or more amino acid substitutions, deletions, or additions (e.g., up to 20, up to 15, up to 10, or up to 5 amino acid substitutions, deletions, or additions, e.g., 1, 2, 3, 4, or 5 amino acid substitutions, deletions, or additions) compared to the wild-type sequence from which it is derived.

[0012] In some embodiments, the heavy chain constant region is an IgG heavy chain constant region, e.g., an IgG1, IgG2, IgG3, or IgG4 heavy chain constant region, e.g., a human IgG1 heavy chain constant region or a human IgG4 heavy chain constant region. In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain constant region (CH) set forth as SEQ ID NO: 43 or a variant thereof, wherein the variant has up to 20 amino acid conservative substitutions (e.g., up to 15, up to 10, or up to 5 amino acid conservative substitutions, e.g., 1, 2, 3, 4, or 5 amino acid conservative substitutions) compared to SEQ ID NO: 43.

[0013] In some embodiments, the light chain constant region comprises a kappa light chain constant region. In some embodiments, the antibody, or antigen-binding region thereof, comprises a light chain constant region (CL) set forth as SEQ ID NO: 44, or a variant thereof, wherein the variant has up to 20 amino acid conservative substitutions (e.g., up to 15, up to 10, or up to 5 amino acid conservative substitutions, e.g., 1, 2, 3, 4, or 5 amino acid conservative substitutions) compared to SEQ ID NO: 44.

[0014] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain constant region (CH) set forth as SEQ ID NO: 43 or 45 and a light chain constant region (CL) set forth as SEQ ID NO: 44.

[0015] In some embodiments, the antibody or antigen-binding fragment thereof (1) a heavy chain comprising a VH sequence set forth in SEQ ID NO: 1 and a heavy chain constant region (CH) set forth in SEQ ID NO: 43, and a light chain comprising a VL sequence set forth in SEQ ID NO: 2 and a light chain constant region (CL) set forth in SEQ ID NO: 44; (2) a heavy chain comprising a VH sequence set forth in SEQ ID NO: 3 and a heavy chain constant region (CH) set forth in SEQ ID NO: 43 or 45, and a light chain comprising a VL sequence set forth in SEQ ID NO: 4 and a light chain constant region (CL) set forth in SEQ ID NO: 44; (3) a heavy chain comprising a VH sequence set forth as SEQ ID NO: 5 and a heavy chain constant region (CH) set forth as SEQ ID NO: 43, and a light chain comprising a VL sequence set forth as SEQ ID NO: 6 and a light chain constant region (CL) set forth in SEQ ID NO: 44; (4) a heavy chain comprising the VH sequence set forth in SEQ ID NO: 7 and the heavy chain constant region (CH) set forth in SEQ ID NO: 43, and a light chain comprising the VL sequence set forth in SEQ ID NO: 8 and the light chain constant region (CL) set forth in SEQ ID NO: 44; or (5) A heavy chain comprising a VH set forth in SEQ ID NO: 9 and a heavy chain constant region (CH) set forth in SEQ ID NO: 43, and a light chain comprising a VL set forth in SEQ ID NO: 10 and a light chain constant region (CL) set forth in SEQ ID NO: 44.

[0016] In the antibody-drug conjugate, the cytotoxic drug may be linked to the antibody or antigen-binding fragment via a linker (e.g., the "MLE" fragment shown in this application).

[0017] In some embodiments, M is [ka] wherein ring A is a 5- to 6-membered aliphatic heterocycle or a 5- to 20-membered aromatic ring system, and the aliphatic heterocycle and aromatic ring system are substituted with oxo (=O), halogen, cyano, amino, carboxyl, mercapto, and C 1~6 alkyl; and M is a single bond, C 1~20 Alkylene, C 2~20 Alkenylene, and C 2~20 alkynylene.

[0018] In some embodiments, M is [ka] wherein ring A is a 5-membered aliphatic heterocycle, a 6-membered aromatic heterocycle, or a polycyclic ring formed by connecting two or more 6-membered aromatic heterocycles and a benzene ring with a single bond, and the aliphatic heterocycle is selected from the group consisting of oxo (=O), halogen, and C 1~4 alkyl, and M is a single bond, C 3~10 Alkylene, C 3~10 Alkenylene, and C 3~10 alkynylene.

[0019] In some embodiments, M is [ka] and ring A is [ka] M1 is selected from a single bond, C 5~8 Alkylene, C 5~8 Alkenylene, and C 5~8 alkynylene.

[0020] In some embodiments, M has the following structure: [ka] is selected from.

[0021] In some embodiments, M is selected from the following structures: [ka]

[0022] In some embodiments, L is C 1~6Alkylene, -N(R')-, carbonyl, -O-, Val, Cit, Phe, Lys, Lys(COCH2CH2(OCH2CH2) s OCH3), D-Val, Leu, Gly, Ala, Asn, Val-Cit, Val-Ala, Val-Lys, Val-Lys(Ac), Phe-Lys, Phe-Lys(Ac), D-Val-Leu-Lys, Gly-Gly-Arg, Ala-Ala-Asn, Ala-Ala-Ala, Val-Lys-Ala, Val-Lys-Gly, Gly-Gly-Gly, Gly-Gly-Phe-Gly (SEQ ID NO: 48), Gly-Phe-Leu-Gly (SEQ ID NO: 49), Gly-Gly-Val-Ala (SEQ ID NO: 50), Gly-Gly-Gly-Gly-Gly (SEQ ID NO: 51), [ka] and R' is selected from the group consisting of hydrogen, C 1~6 alkyl, or alkyl containing -(CH2CH2O)r-, where r is selected from an integer of 1 to 10, and s is selected from an integer of 1 to 20.

[0023] In some embodiments, L is C 1~6 Alkylene, -N(R')-, carbonyl, -O-, Val, Cit, Phe, Lys, Lys(COCH2CH2(OCH2CH2) s OCH3), D-Val, Leu, Gly, Ala, Asn, Val-Cit, Val-Ala, Val-Lys, Val-Lys(Ac), Phe-Lys, Phe-Lys(Ac), D-Val-Leu-Lys, Gly-Gly-Arg, Ala-Ala-Asn, Ala-Ala-Ala, Val-Lys-Ala, Val-Lys-Gly, Gly-Gly-Gly, Gly-Gly-Phe-Gly (SEQ ID NO: 48), Gly-Phe-Leu-Gly (SEQ ID NO: 49), Gly-Gly-Val-Ala (SEQ ID NO: 50), Gly-Gly-Gly-Gly-Gly (SEQ ID NO: 51), [ka] wherein R' represents hydrogen, C1-6 alkyl, or alkyl containing -(CH2CH2O)r-, r is selected from an integer of 1 to 10, and s is selected from an integer of 1 to 20.

[0024] In some embodiments, L is C 1~6 Alkylene, -NH-, Val, Cit, Phe, Lys, Lys(COCH2CH2(OCH2CH2) s OCH3), Gly, Val-Cit, Gly-Gly-Phe-Gly (SEQ ID NO: 48), [ka] and s is selected from an integer of 1 to 20.

[0025] In some embodiments, L is C 1~6 Alkylene, -NH-, Val, Cit, Phe, Lys, Lys(COCH2CH2(OCH2CH2) s OCH3), Gly, Val-Cit, Gly-Gly-Phe-Gly (SEQ ID NO: 48), [ka] and s is selected from an integer of 1 to 20.

[0026] In some embodiments, L has the following structure: [ka] is selected from the group consisting of s is selected from integers of 1 to 20. In some embodiments, L has the following structure: [ka] is selected from s is selected from integers of 1 to 20.

[0027] In some embodiments, L has the following structure: [ka] is selected from.

[0028] In some embodiments, L has the following structure: [ka] is selected from.

[0029] In some embodiments, L has the following structure: [ka] is selected from.

[0030] In some embodiments, L has the following structure: [ka] is selected from.

[0031] In some embodiments, E is a single bond, —NH—CH 2 —, —NH—CH 2 —O—CH 2 —CO—, [ka] is.

[0032] In some embodiments, E is a single bond, —NH—CH 2 —, —NH—CH 2 —O—CH 2 —CO—, [ka] is.

[0033] In some embodiments, E is -NH-CH2-, [ka] is.

[0034] In some embodiments, E is -NH-CH2- or [ka] is.

[0035] In some embodiments, M has the following structure: [ka] is selected from L has the following structure: [ka] is selected from E is -NH-CH2, [ka] is.

[0036] In some embodiments, M has the following structure: [ka] is selected from L has the following structure: [ka] is selected from E is -NH-CH2, [ka] is.

[0037] In some embodiments, [ka] is selected from the following structures: [ka] [ka] [ka] [ka]

[0038] In some embodiments, [ka] is selected from the following structures: [ka] [ka] [ka]

[0039] In some embodiments, [ka] is selected from the following structures: [ka] , [ka] , [ka] , [ka] , [ka] , [ka] , [ka]

[0040] In some embodiments, [ka] is selected from the following structures: [ka]

[0041] In some embodiments, the cytotoxic drug is selected from a tubulin inhibitor, a DNA intercalator, a DNA topoisomerase inhibitor, and an RNA polymerase inhibitor. In some embodiments, the tubulin inhibitor is an auristatin compound or a maytansinoid compound. In some embodiments, the DNA intercalator is a pyrrolobenzodiazepine (PBD). In some embodiments, the DNA topoisomerase inhibitor is a topoisomerase I inhibitor (e.g., camptothecin, hydroxycamptothecin, 9-aminocamptothecin, SN-38, irinotecan, topotecan, belotecan, or rubitecan) or a topoisomerase II inhibitor (e.g., doxorubicin, PNU-159682, duocarmycin, daunorubicin, mitoxantrone, podophyllotoxin, or etoposide). In some embodiments, the RNA polymerase inhibitor is α-amanitin or a pharmaceutically acceptable salt, ester, or analog thereof.

[0042] Cytotoxic drugs disclosed in this application generally contain hydroxyl (-OH), carboxyl (-COOH), sulfhydryl (-SH), primary amino (-NH), secondary amine groups (-NR A H), or a tertiary amine group (-NR B R C ), and R A , RB , and R C represents only non-hydrogen substituents on the N, and the cytotoxic drug can be linked to the linker in the conjugate via these functional groups.

[0043] In some embodiments, the cytotoxic agent is linked to E in the antibody-drug conjugate via -OH, -SH, a primary amino group, a secondary amine group, or a tertiary amine group.

[0044] In some embodiments, the cytotoxic agent is [ka] is.

[0045] In some embodiments, the cytotoxic agent is selected from Formulas I and II: [ka] R1 and R2 each independently represent C 1~6 selected from alkyl and halogen; R3 is selected from H and —CO—CH2OH; R4 and R5 are each independently selected from H, halogen, and hydroxyl, or R4 and R5 are joined to associated carbon atoms to form a 5- to 6-membered oxygen-containing heterocycle; R6 is hydrogen and -C 1~4 Alkylene-NR a R b is selected from R7 is hydrogen, C 1~6 Alkyl, and -C 1~4 Alkylene-NR a R b is selected from In each occurrence, R a and R b are each independently H, C 1~6 Alkyl, -SO2-C 1~6 Alkyl, and -CO-C 1~6 alkyl.

[0046] In some embodiments, the cytotoxic agent is selected from Formulas I and II: [ka] R1 and R2 each independently represent C 1~6 selected from alkyl and halogen; R3 is selected from H and —CO—CH2OH; R4 and R5 are each independently selected from H, halogen, and hydroxyl, or R4 and R5 are joined to associated carbon atoms to form a 5- to 6-membered oxygen-containing heterocycle; R6 is hydrogen and -C 1~4 Alkylene-NR a R b is selected from R7 is C 1~6 Alkyl and -C 1~4 Alkylene-NR a R b is selected from In each occurrence, R a and R b are each independently H, C 1~6 Alkyl, -SO2-C 1~6 Alkyl, and -CO-C 1~6 alkyl.

[0047] In some embodiments, the cytotoxic agent has the following structure: [ka] is selected from.

[0048] In some embodiments, the cytotoxic agent has the following structure: [ka] is selected from.

[0049] The corresponding fragment of the cytotoxic drug obtained after the cytotoxic drug is linked to the linker has the formula Ab-[MLED] according to the present application. xIn some embodiments, D is a monovalent structure obtained by losing one H from an -OH, -NH, or secondary amine group on the cytotoxic drug.

[0050] In some embodiments, D has the following structure: [ka] is selected from.

[0051] In some embodiments, the antibody-drug conjugate is selected from ADC A-01 to ADC A-26, ADC B-01 to ADC B-06, and ADC C-01 shown below. [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] HA is any one of the antibodies disclosed herein; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and the linker.

[0052] In some embodiments, the antibody-drug conjugates are ADC A-01 to ADC A-25, ADC B-01 to ADC B-06, and ADC C-01 shown below: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] HA is any one of the antibodies disclosed herein; [ka] represents the specific linkage between the sulfhydryl in the antibody or antigen-binding fragment and the linker.

[0053] In certain embodiments, the specific linking mode is a linkage or bond formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker.

[0054] In a further embodiment, the HA is (1a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 11, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 12, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13; and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16; (1b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 27, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 28, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29; and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32; (2a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 17, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 18 or 19, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13; and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16; (2b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 33, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 34 or 35, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29; and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32; (3a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 20, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 21, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 22; and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 23, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 24, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16; (3b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 36, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 37, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 38; and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 39, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 40, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32; (4a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 25, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 26, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13; and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16; or (4b) An antibody comprising a heavy chain variable region (VH) including the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 41, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 42, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29, and / or a light chain variable region (VL) including the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32.

[0055] In a further embodiment, the HA comprises a VH having the amino acid sequence set forth in SEQ ID NO:1 and / or a VL having the amino acid sequence set forth in SEQ ID NO:2.

[0056] In a further embodiment, the HA comprises a VH having the amino acid sequence set forth in SEQ ID NO:3 and / or a VL having the amino acid sequence set forth in SEQ ID NO:4.

[0057] In a further embodiment, the HA comprises a VH having the amino acid sequence set forth in SEQ ID NO:5 and / or a VL having the amino acid sequence set forth in SEQ ID NO:6.

[0058] In a further embodiment, the HA comprises a VH having the amino acid sequence set forth in SEQ ID NO:7 and / or a VL having the amino acid sequence set forth in SEQ ID NO:8.

[0059] In a further embodiment, the HA comprises a VH having the amino acid sequence set forth in SEQ ID NO:9 and / or a VL having the amino acid sequence set forth in SEQ ID NO:10.

[0060] In a further embodiment, the HA comprises (1) a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43, and a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44.

[0061] In a further embodiment, the HA comprises (2) a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44.

[0062] In a further embodiment, the HA comprises (3) a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43, and a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44.

[0063] In a further embodiment, the HA comprises (4) a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO:7 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO:43, and a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO:8 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO:44.

[0064] In a further embodiment, the HA comprises (5) a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43, and a light chain comprising a VL sequence having the amino acid sequence set forth in SEQ ID NO: 10 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44.

[0065] In a further embodiment, the HA comprises a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO:46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO:47.

[0066] In some embodiments, the antibody or antigen-binding fragment thereof (a) a human immunoglobulin heavy chain constant region (CH) or a variant thereof, wherein the variant has one or more amino acid substitutions, deletions, or additions (e.g., up to 20, up to 15, up to 10, or up to 5 amino acid substitutions, deletions, or additions, e.g., 1, 2, 3, 4, or 5 amino acid substitutions, deletions, or additions) compared to the wild-type sequence from which it is derived; and (b) A human immunoglobulin light chain constant region (CL) or variant thereof, wherein the variant further comprises one or more amino acid substitutions, deletions, or additions (e.g., up to 20, up to 15, up to 10, or up to 5 amino acid substitutions, deletions, or additions, e.g., 1, 2, 3, 4, or 5 amino acid substitutions, deletions, or additions) compared to the wild-type sequence from which it is derived.

[0067] In one embodiment, the HA in each antibody-drug conjugate represents an antibody or antigen-binding fragment of a VH set forth as SEQ ID NO: 1, 3, 5, 7, or 9 and a VL set forth as SEQ ID NO: 2, 4, 6, 8, or 10, e.g., an antibody or antigen-binding fragment of a VH set forth as SEQ ID NO: 3 and a CH set forth as SEQ ID NO: 43 or 45, a VL set forth as SEQ ID NO: 4 and a CL set forth as SEQ ID NO: 44, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and the linker.

[0068] In certain embodiments of the HA or antibody-drug conjugates disclosed herein, the heavy chain constant domain may include a C-terminal lysine or may lack either a C-terminal lysine or a C-terminal glycine-lysine dipeptide. In some embodiments of the antibody or antigen-binding fragment thereof, the N-terminal amino acid of the antibody or antigen-binding fragment thereof variable domain may undergo cyclization to pyroglutamate.

[0069] In some embodiments of the antibody or antigen-binding fragment thereof, the N-terminal amino acid of the antibody or antigen-binding fragment thereof variable domain may undergo cyclization to pyroglutamic acid.

[0070] Thus, a composition can comprise a population of antibody-drug conjugate species, each of which can independently comprise a C-terminal lysine, lack a C-terminal lysine, lack a C-terminal glycine-lysine, and / or comprise an N-terminal glutamine or glutamic acid or cyclization of the N-terminal amino acid to pyroglutamate.

[0071] In certain embodiments, a composition may comprise a population of antibody-drug conjugate species, each of which may independently comprise a C-terminal lysine, lack a C-terminal lysine, lack a C-terminal glycine-lysine, and / or comprise an N-terminal glutamine or glutamic acid or cyclization of the N-terminal amino acid to pyroglutamic acid.

[0072] Accordingly, in certain embodiments, the present invention further provides a composition comprising an ADC disclosed herein, wherein the predominant ADC species in the composition comprises (i) an antibody whose heavy chain C-terminus lacks a lysine residue, (ii) an antibody whose heavy chain N-terminus is glutamine, glutamic acid, or pyroglutamate, or (iii) an antibody whose heavy chain C-terminus lacks a lysine residue and whose heavy chain N-terminus is glutamine, glutamic acid, or pyroglutamate.

[0073] In certain embodiments, the invention further provides a composition comprising an ADC disclosed herein, wherein the predominant ADC species in the composition comprises (i) an antibody whose heavy chain C-terminus lacks a lysine residue, (ii) an antibody whose heavy chain N-terminus is glutamine, glutamic acid, or pyroglutamic acid, or (iii) an antibody whose heavy chain C-terminus lacks a lysine residue and whose heavy chain N-terminus is glutamine, glutamic acid, or pyroglutamic acid.

[0074] As known to those skilled in the art, pyroglutamic acid is the conjugate acid of pyroglutamate and is in equilibrium with pyroglutamate in solution.

[0075] composition In another aspect, the present application provides compositions of antibody-drug conjugates (ADCs) described herein. Such compositions may comprise a plurality of ADCs described herein, each ADC comprising a drug-linker described herein. Thus, compositions can be characterized by a "drug-to-antibody" ratio (DAR) ranging from about 1 to about 10. Methods for determining the DAR are well known to those of skill in the art and include methods using reverse-phase chromatography or HPLC-MS.

[0076] In some embodiments, the DAR value (drug-antibody conjugate ratio) of the composition of drug-antibody conjugate is 1 to 10, e.g., 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, 1 to 10, 2 to 3, 2 to 4, 2 to 5, 2 to 6, 2 to 7, 2 to 8, 2 to 9, 2 to 10, 3 to 4, 3 to 5, 3 to 6, 3 to 7, 3 to 8, 3 to 9, 3 to 10, 4 to 5, 4 to 6, 4 to 7, 4 to 8, 4 to 9, 4 to 10, 5 to 6, 5 to 7, 5 to 8, 5 to 9, 5 to 10, 6 to 7, 6 to 8, 6 to 9, 6 to 10, 7 to 8, 7 to 9, 7 to 10, 8 to 9, 8 to 10, or 9 to 10, preferably 1 to 8, preferably 3 to 9, for example, 3.0 to 3.5, 3.0 to 4.0, 3.0 to 4.5, 3.0 to 5.0, 3.0 to 5.5, 3.0 to 6.0, 3.5 to 4.0, 3.5 to 4.5, 3.5 to 5.0, 3. 5~5.5, 3.5~6.0, 3.5~6.5, 3.5~7.0, 3.5~7.5, 3.5~8.0, 4.0~4.5, 4.0~5.0, 4.0~5.5, 4.0~6.0, 4.0~6.5, 4.0~7.0, 4.0~7.5, 4.0~8.0, 4.5~5.0, 4.5~5.5, 4.5~6.0, 4.5~6.5, 4.5~7.0, 4.5~7.5, 4.5~8.0, 5.0 ~5.5, 5.0~6.0, 5.0~6.5, 5.0~7.0, 5.0~7.5, 5.0~8.0, 5.5~6.0, 5.5~6.5, 5.5~7.0, 5.5~7.5, 5.5~8.0, 6.0~6.5, 6.0~7.0, 6.0~7.5, 6.0~8.5, 6.5~7.0, 6.5~7.5, 6.5~8.5, 7.0~7.5, 7.0~9.0, or 7.5~9.0.

[0077] In one embodiment, the DAR value is 8. In one embodiment, the DAR value is 7 to 9. In one embodiment, the DAR value is 7 to 8. In another embodiment, the DAR value of an antibody-drug conjugate composition comprising ADC A-05 is about 8. In another embodiment, the DAR value of an antibody-drug conjugate composition comprising ADC A-05 is 7 to 9. In another embodiment, the DAR value of an antibody-drug conjugate composition comprising ADC A-05 is 7 to 8. In another embodiment, the DAR value of an antibody-drug conjugate composition comprising ADC B-01 is 7 to 8. In another embodiment, the DAR value of an antibody-drug conjugate composition comprising ADC C-01 is 2 to 4. In another embodiment, the DAR value of an antibody-drug conjugate composition comprising ADC B-04 is 7 to 9. In another embodiment, the DAR value of an antibody-drug conjugate composition comprising ADC A-26 is 7 to 9.

[0078] Drug-Linker Those skilled in the art should understand that the antibody-drug conjugates (ADCs) described herein can be prepared in a modular manner. For example, a free form of a "drug-linker" (which can be understood as M'-L-E, where M' is the structural form of M prior to covalent attachment to an antibody or antigen-binding fragment thereof) can be obtained, and then the free form of the "drug-linker" can be covalently attached to an antibody or antigen-binding fragment thereof to obtain an antibody-drug conjugate according to the present application. Correspondingly, M' in the free form of the "drug-linker" can be linked to one or more sulfhydryls (-SH), amino (-NH), or carboxyls (-COOH) on an antibody or antigen-binding fragment thereof via substitution reactions (e.g., removal of -SOMe or -Br and similar structures thereon) or addition reactions and other methods.

[0079] In another embodiment, the present invention provides a Drug-Linker having a structure shown as formula M'-LED, M' is [ka] and Lg is a leaving group for nucleophilic substitution reactions (e.g., halogen, methanesulfonyl, fluorophenol, or [ka] ) or hydroxyl (-OH), sulfhydryl group (-SH), or amino (-NH2), or Lg forms an unsaturated double bond with adjacent atoms on ring A, ring A is a 5- to 6-membered aliphatic heterocycle or a 5- to 20-membered aromatic ring system, and the aliphatic heterocycle and aromatic ring system are oxo (=O), halogen, cyano, amino, carboxyl, sulfhydryl, and C 1~6 alkyl, and M is a single bond, C 1~20 Alkylene, C 2~20 Alkenylene, and C 2~20 alkynylene, The L, E, and D structures are defined according to any one of the above antibody-drug conjugates.

[0080] In some embodiments, M' is [ka] Lg is methylsulfonyl, or Lg forms a carbon-carbon double bond together with adjacent atoms on ring A, ring A is a 5-membered alicyclic heterocycle, a 6-membered heteroaromatic ring, or a polycyclic ring formed by connecting two or more 6-membered heteroaromatic rings and a benzene ring with a single bond, and the alicyclic heterocycle is substituted with oxo (=O), halogen, and C 1~4 alkyl, and M is a single bond, C 3~10 Alkylene, C 3~10 Alkenylene, and C 3~10 alkynylene.

[0081] In some embodiments, M' is [ka] and [ka] M1 is selected from a single bond, C 5~8 Alkylene, C 5~8 Alkenylene, and C 5~8 alkynylene.

[0082] In some embodiments, M' is [ka] is selected from the group consisting of:

[0083] In some embodiments, M' is [ka] is.

[0084] In some embodiments, the free form of the "drug-linker" is selected from A-01 to A-26, B-01 to B-06, and C-01 shown below. [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka]

[0085] In some embodiments, the free form of the "drug-linker" is selected from A-01 to A-25, B-01 to B-06, and C-01 shown below. [ka] [ka] [ka] [ka] [ka] [ka] [ka]

[0086] Pharmaceutical Composition In another aspect, the present application provides a pharmaceutical composition comprising an antibody-drug conjugate described in any one of the preceding paragraphs and optionally a drug-linker described in any one of the preceding paragraphs, and one or more pharmaceutically acceptable adjuvants or agents.

[0087] The antibody-drug conjugates described herein are typically formulated in a unit injectable form together with a pharmaceutically acceptable parenteral vehicle for parenteral use, e.g., bolus injection, intravenous injection, intratumoral injection, etc. Optionally, the antibody-drug conjugates having the desired purity are mixed with a pharmaceutically acceptable diluent, carrier, excipient, or stabilizer in the form of a lyophilizate or solution (Remington's Pharmaceutical Sciences (1980) 16). th edition, Osol, A. Ed.) The antibody-drug conjugates, or pharmaceutical compositions comprising the antibody-drug conjugates described herein, can be administered via any route appropriate to the individual being treated.

[0088] use The antibody-drug conjugates or pharmaceutical compositions thereof described herein can be used to treat various diseases or disorders, such as cancers associated with high expression of PTK7, including solid tumors or hematological malignancies, e.g., lung cancer, breast cancer, epidermal cancer (e.g., squamous cell carcinoma of the skin and oral squamous cell carcinoma), ovarian cancer, esophageal cancer (e.g., squamous cell carcinoma of the esophagus), etc.

[0089] Thus, the present application provides the use of an antibody-drug conjugate, a drug-linker, or a pharmaceutical composition comprising the same in the preparation of a medicament for treating a cancer associated with high expression of PTK7.

[0090] Meanwhile, the present application also provides a method for treating cancer associated with high expression of PTK7, comprising administering to a subject in need thereof an effective amount of any one of the antibody-drug conjugates, drug linkers, or pharmaceutical compositions comprising them described above. In certain embodiments, the cancer comprises a solid tumor or a hematological malignancy. In further embodiments, the cancer is lung cancer, breast cancer, epidermal cancer (e.g., cutaneous squamous cell carcinoma and oral squamous cell carcinoma), ovarian cancer, esophageal cancer (e.g., esophageal squamous cell carcinoma), etc. [Brief explanation of the drawings]

[0091] [Figure 1A] Detection of the efficacy of various antibody-drug conjugates in the NCI-H358 model.

[0092] [Figure 1B] Body weight detection of various antibody-drug conjugates in the NCI-H358 model.

[0093] [Figure 2A] Efficacy detection of various antibody-drug conjugates in the A431 model.

[0094] [Figure 2B] Body weight detection of various antibody-drug conjugates in the A431 model.

[0095] [Figure 3A] Detection of binding of anti-human PTK7 antibody-drug conjugates to OVCAR3 cells.

[0096] [Figure 3B] Detection of binding of anti-human PTK7 antibody-drug conjugates to NCI-H358 cells.

[0097] [Figure 3C] Detection of binding of anti-human PTK7 antibody-drug conjugates to HCC1806 cells.

[0098] [Figure 3D] Detection of binding of anti-human PTK7 antibody-drug conjugates to A431 cells.

[0099] [Figure 3E] Detection of binding of anti-human PTK7 antibody-drug conjugates to NCI-H520 cells.

[0100] [Figure 3F] Detection of binding of anti-human PTK7 antibody-drug conjugates to DU4475 cells.

[0101] [Figure 4A] pHrodo detection of endocytosis of antibody-drug conjugates in NCI-H358.

[0102] [Figure 4B] pHrodo detection of endocytosis of antibody-drug conjugates in HCC1806.

[0103] [Figure 4C] pHrodo detection of endocytosis of antibody-drug conjugates in OVCAR3.

[0104] [Figure 4D] pHrodo detection of endocytosis of antibody-drug conjugates in NCI-H520.

[0105] [Figure 4E] pHrodo detection of endocytosis of antibody-drug conjugates in DU4475.

[0106] [Figure 5A] Detection of anti-human PTK7-bound drugs in killing of FADU cells.

[0107] [Figure 5B] Detection of anti-human PTK7-bound drugs in killing of HCC1806 cells.

[0108] [Figure 5C] Detection of anti-human PTK7-bound drugs in killing of DU4475 cells.

[0109] [Figure 6A] Detection of the efficacy of antibody-drug conjugates in the HCC1806 model.

[0110] [Figure 6B] Body weight detection of antibody-drug conjugates in the HCC1806 model.

[0111] [Figure 7A] Detection of antibody-drug conjugate efficacy in the OVCAR3 model.

[0112] [Figure 7B] Body weight detection of antibody-drug conjugates in the OVCAR3 model.

[0113] [Figure 8A] Detection of antibody-drug conjugate efficacy in the NCI-H146 model.

[0114] [Figure 8B] Body weight detection of antibody-drug conjugates in the NCI-H146 model. DETAILED DESCRIPTION OF THE INVENTION

[0115] definition Unless otherwise defined hereinafter, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. References to techniques used herein are intended to refer to techniques commonly understood in the art, including modifications of the techniques or substitutions of equivalent techniques that are obvious to those skilled in the art. Furthermore, the laboratory procedures of genomics, nucleic acid chemistry, and molecular biology used herein are all routine procedures widely used in the corresponding fields. While the following terms are believed to be well understood by those skilled in the art, the following definitions are provided for a better explanation of the present invention.

[0116] The term "antibody" typically refers to an immunoglobulin molecule composed of two pairs of polypeptide chains, each pair having a light chain (LC) and a heavy chain (HC). Antibody light chains can be classified as kappa (κ) and lambda (λ) light chains. Heavy chains can be classified as μ, δ, γ, α, or ε, and antibody isotypes can be defined as IgM, IgD, IgG, IgA, and IgE, respectively. Within light and heavy chains, the variable and constant regions are connected by a "J" region of about 12 or more amino acids, and heavy chains may further include a "D" region of about 3 or more amino acids. Each heavy chain is composed of a heavy chain variable region (VH) and a heavy chain constant region (CH). The heavy chain constant region is composed of three domains (CH1, CH2, and CH3). Each light chain is composed of a light chain variable region (VL) and a light chain constant region (CL). The light chain constant region is composed of one domain, CL. The constant domains are not directly involved in antibody and antigen binding, but exhibit various effector functions, such as mediating the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component of the classical complement system (C1q). The VH and VL regions can also be further divided into highly variable regions (called complementarity-determining regions (CDRs)) interspersed with more conserved regions called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs, arranged in the following order from amino terminus to carboxy terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions (VH and VL) of each heavy / light chain pair form the respective antigen-binding site. The assignment of amino acids to each region or domain may follow various numbering systems known in the art.

[0117] The term "complementarity-determining region" or "CDR" refers to the amino acid residues in the variable region of an antibody that are involved in antigen binding. Each of the heavy and light chain variable regions contains three CDRs, designated CDR1, CDR2, and CDR3. The exact boundaries of these CDRs can be determined according to various numbering systems known in the art, such as the Kabat numbering system (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991), the Chothia numbering system (Chothia & Lesk (1987) J. Mol. Biol. 196:901-917, Chothia et al. (1989) Nature 342:878-883), the IMGT numbering system (Lefranc et al., Dev. Compare. Immunol. 27:55-77, 2003), or the AbM numbering system (Martin ACR, Cheetham JC, Rees AR (1989) Modeling antibody hypervariable loops: A combined algorithm. Proc Natl Acad Sci USA 86:9268-9272). For a given antibody, one of skill in the art would readily identify the CDRs defined according to each numbering system. Furthermore, the correspondence between different numbering systems is well known to those of skill in the art (see, e.g., Lefranc et al., Dev. Comparat. Immunol. 27:55-77, 2003).

[0118] In the present invention, the CDRs contained in the antibody or antigen-binding fragment thereof of the present invention can be determined according to various numbering systems known in the art, such as the Kabat, Chothia, IMGT, or AbM numbering systems. In certain embodiments, the CDRs contained in the antibody or antigen-binding fragment thereof are determined according to the Chothia numbering system.

[0119] The following general rules, disclosed in www.bioinf.org.uk: Prof. Andrew C. Martin's Group and reproduced below, can be used to define CDRs in an antibody sequence that contain amino acids that interact specifically with amino acids that comprise the epitope in the antigen to which the antibody binds. There are rare instances where these generally consistent features do not occur; however, Cys residues are the most conserved features. [Table 1]

[0120] V H The entire amino acid sequence of V is generally numbered according to Kabat, although the three CDRs within the variable region may be defined according to any one of the aforementioned numbering schemes. H The numbering of amino acid positions in a sequence may start at amino acid position 1 and continue consecutively to the end of the sequence, or may be consecutive according to Kabat. H and V L The amino acid positions within are defined according to consecutive numbering.

[0121] The numbering of amino acid positions in the heavy chain constant domain can start at amino acid position 1 and continue consecutively to the end of the sequence, or can be consecutive according to Eu numbering. The IgG1 heavy chain constant domain amino acid sequence has 330 amino acids numbered consecutively from 1 to 330. The corresponding sequence numbered according to Eu starts at position 118 and ends at position 447. Unless otherwise specified, the amino acid positions in the heavy and light chains herein are defined according to consecutive numbering.

[0122] The term "framework region" or "FR" residues refers to amino acid residues in antibody variable regions other than the CDR residues as defined above.

[0123] The term "antigen-binding fragment" of an antibody refers to a polypeptide fragment of an antibody, e.g., a polypeptide fragment of a full-length antibody, which retains the ability to specifically bind to the same antigen bound by the full-length antibody and / or competes with the full-length antibody for specific binding to an antigen, also referred to as an "antigen-binding fragment." Antigen-binding fragments are generally described in Fundamental Immunology, Ch. 7 (Paul, W., ed., 2nd ed., Raven Press, NY (1989)), which is incorporated herein by reference in its entirety for all purposes. Antigen-binding fragments of antibodies can be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies. Non-limiting examples of antigen-binding fragments include Fab fragments, Fab' fragments, F(ab')2 fragments, F(ab')3 fragments, Fd, Fv, scFv, di-scFv, (scFv)2, disulfide bond stabilized Fv proteins ("dsFv"), single domain antibodies (sdAb, nanobodies), and other such polypeptides that contain at least a sufficient portion of an antibody to confer specific antigen-binding ability to the polypeptide. Engineered variants of antibodies are reviewed in Holliger et al., 2005; Nat Immunol 23, 1126-1136.

[0124] The term "Fd" refers to an antibody fragment consisting of the VH and CH1 domains, the term "dAb fragment" refers to an antibody fragment consisting of the VH domain (Ward et al., Nature 341:544 546 (1989)), the term "Fab fragment" refers to an antibody fragment consisting of the VL, VH, CL, and CH1 domains, the term "F(ab')2 fragment" refers to an antibody fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region, and the term "Fab' fragment" refers to a fragment obtained after reducing the disulfide bond linking the two heavy chain fragments of the F(ab')2 fragment, wherein the resulting fragment consists of a light chain and a complete Fd fragment (consisting of the VH and CH1 domains) of a heavy chain.

[0125] The term "Fv" refers to an antibody fragment consisting of the VL and VH domains of a single arm of an antibody. An Fv fragment is generally considered to be the smallest antibody fragment capable of forming a complete antigen-binding site. It is generally believed that six CDRs can confer antigen-binding specificity to an antibody. However, a single variable region (e.g., an Fd fragment containing only three antigen-specific CDRs) can also be used to recognize and bind to an antigen, although the affinity may be lower than that of the complete binding site.

[0126] The term "Fc fragment" refers to an antibody fragment formed by linking the second and third constant regions of a first heavy chain of an antibody to the second and third constant regions of a second heavy chain via disulfide bonds. The Fc fragment of an antibody has a variety of different functions, but is not involved in antigen binding.

[0127] The term "scFv" refers to a single polypeptide chain comprising a VL and a VH domain, wherein the VL and VH are connected by a linker (see, e.g., Bird et al., Science 242:423-426 (1988); Huston et al., Proc. Natl. Acad. Sci. USA 85:5879-5883 (1988); and Plückthun, The Pharmacology of Monoclonal Antibodies, Volume 113, edited by Roseburg and Moore, Springer-Verlag, New York, pp. 269-315 (1994)). Such scFv molecules can have the general structure: NH2-linker-VH-COOH or NH2-VH-linker-VL-COOH. Suitable linkers in the prior art consist of repeated GGGGS (SEQ ID NO: 55) amino acid sequences or variants thereof. For example, a linker having the amino acid sequence (GGGGS)4 (SEQ ID NO: 56) can be used, although variants thereof can also be used (Holliger et al. (1993), Proc. Natl. Acad. Sci. USA 90:6444-6448). Other linkers applicable to the present invention are described by Alfthan et al. (1995), Protein Eng. 8:725-731; Choi et al. (2001), Eur. J. Immunol. 31:94-106; Hu et al. (1996), Cancer Res. 56:3055-3061; Kipriyanov et al. (1999), J. Mol. Biol. 293:41-56; and Roovers et al. (2001), Cancer Immunol. In some cases, a disulfide bond may exist between the VL and VH of the scFv. In certain embodiments, the VH and VL domains may be positioned opposite each other in any suitable arrangement, e.g., NH2-VH-VH-COOH and NH 2- scFv containing VL-VL-COOH.

[0128] The term "single domain antibody (sdAb)" has the meaning commonly understood by those skilled in the art and refers to an antibody fragment composed of a single monomeric variable antibody domain (e.g., a single heavy chain variable region), which antibody fragment retains the ability to specifically bind to the same antigen as the full-length antibody (Holt, L. et al., Trends in Biotechnology, 21(11): 484-490, 2003). Single domain antibodies are also called nanobodies.

[0129] Each of the above antibody fragments retains the ability to specifically bind to the same antigen bound by the full-length antibody and / or competes with the full-length antibody for specific binding to the antigen.

[0130] In this specification, unless otherwise specified, reference to the term "antibody" includes not only complete antibodies but also antigen-binding fragments of antibodies.

[0131] Antigen-binding fragments of antibodies (e.g., the antibody fragments described above) can be obtained from a given antibody (such as an antibody provided by the present invention) using conventional techniques known to those of skill in the art (e.g., recombinant DNA techniques or enzymatic or chemical cleavage methods), and the antigen-binding fragments of an antibody are screened for specificity in the same manner as intact antibodies.

[0132] The term "mouse antibody" refers to an antibody obtained by the following method: fusing B cells of an immunized mouse with myeloma cells, screening for mouse hybrid fusion cells that can immortalize and secrete the antibody, followed by screening, antibody preparation, and antibody purification; or refers to an antibody secreted by plasma cells formed by B cell differentiation and proliferation after antigen entry into the mouse body.

[0133] The term "humanized antibody" refers to a genetically engineered non-human antibody whose amino acid sequence has been modified to increase homology to the sequence of a human antibody. Generally, all or a portion of the CDR regions of a humanized antibody are derived from a non-human antibody (donor antibody), and all or a portion of the non-CDR regions (e.g., variable region FRs and / or constant region) are derived from a human immunoglobulin (acceptor antibody). Humanized antibodies generally retain the expected properties of the donor antibody, including, but not limited to, antigen specificity, affinity, reactivity, immune cell activity, ability to enhance immune cell activity, and ability to enhance an immune response. The donor antibody can be a mouse, rat, rabbit, or non-human primate (such as a cynomolgus monkey) antibody with the desired properties (e.g., antigen specificity, affinity, reactivity, ability to enhance immune cell activity, and / or ability to enhance an immune response).

[0134] As used herein, the term "identity" refers to the degree of sequence identity between two polypeptides or two nucleic acids. If a position in two compared sequences is occupied by the same base or amino acid monomer subunit (e.g., if a position in each of two DNA molecules is occupied by adenine, or if a position in each of two polypeptides is occupied by lysine), then the molecules are identical at that position. The "percent identity" between two sequences refers to a function obtained by the following formula: the number of matched positions common to the two sequences / the number of positions compared x 100. For example, if there are 6 matches out of 10 positions in two sequences, the two sequences have 60% identity. For example, the DNA sequences CTGACT and CAGGTT have a total identity of 50% (3 matches out of 6 positions). Typically, comparison is performed when the two sequences are aligned to achieve maximum identity. Such alignment can be achieved, for example, by using the method proposed by Needleman et al. (1970) J. Mol. Biol. 48:443-453 and conveniently implemented by computer programs such as the Align program (DNAstar, Inc.). The percent identity between two amino acid sequences can also be determined using the algorithm of E. Meyers and W. Miller (Comput. Appl. Biosci. Appl. Biosci., 4:11-17 (1988)), which has been incorporated into the ALIGN program (version 2.0) using a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4. Furthermore, the percent identity between two amino acid sequences can be determined by the Needleman and Wunsch (J MoI Biol. 48:444-453 (1970)) algorithm in the GAP program incorporated into the GCG software package (available at www.gcg.com) by using a Blossum 62 matrix or a PAM250 matrix and gap weights of 16, 14, 12, 10, 8, 6, or 4 and length weights of 1, 2, 3, 4, 5, or 6.

[0135] The term "conservative substitution" refers to an amino acid substitution that does not adversely affect or change the expected properties of a protein / polypeptide having the amino acid sequence. For example, conservative substitutions can be introduced by standard techniques known in the art (e.g., site-directed mutagenesis and PCR-mediated mutagenesis). Conservative amino acid substitutions include the replacement of an amino acid residue with an amino acid residue having a similar side chain, e.g., with an amino acid residue that is physically or functionally similar to the corresponding amino acid residue (e.g., has chemical properties including similar size, shape, charge, ability to form covalent or hydrogen bonds, etc.). Families of amino acid residues with similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, and histidine). Accordingly, corresponding amino acid residues are preferably replaced with another amino acid residue from the same side chain family. Methods for identifying conservative amino acid substitutions are well known in the art (see, e.g., Brummell et al., Biochem. 32:1180-1187 (1993); Kobayashi et al., Protein Eng. 12(10): 879-884 (1999); and Burks et al., Proc. Natl. Acad. Set USA 94:412-417 (1997), which are incorporated herein by reference).

[0136] The compilation of the 20 conventional amino acids included herein follows conventional usage. See, for example, Immunology-A Synthesis (2nd Edition, E.S. Golub and D.R. Gren, Eds., Sinauer Associates, Sunderland, Mass. (1991)), which is incorporated herein by reference. In the present invention, amino acids are generally represented by one-letter and three-letter abbreviations well known in the art. For example, alanine can be represented by A or Ala.

[0137] The term "pharmaceutically acceptable carrier and / or excipient" refers to a carrier and / or excipient that is pharmacologically and / or physiologically compatible with the subject and active ingredient, and is well known in the art (see, e.g., Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995). Examples of suitable carriers include, but are not limited to, pH adjusters, surfactants, adjuvants, ionic strength enhancers, diluents, agents for maintaining osmotic pressure, agents for delaying absorption, and preservatives. For example, pH adjusters include, but are not limited to, phosphate buffers. Surfactants include, but are not limited to, cationic, anionic, or nonionic surfactants such as Tween-80. Ionic strength enhancers include, but are not limited to, sodium chloride. Preservatives include, but are not limited to, various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, and sorbic acid. Agents for maintaining osmotic pressure include, but are not limited to, sugars, NaCl, and the like. Agents that delay absorption include, but are not limited to, monostearate and gelatin. Diluents include, but are not limited to, water, aqueous buffer solutions (e.g., buffered saline), alcohols, and polyols (e.g., glycerol). Preservatives include, but are not limited to, various antibacterial and antifungal agents, such as thimerosal, 2-phenoxyethanol, parabens, chlorobutanol, phenol, and sorbic acid. Stabilizers have the meaning commonly understood by those skilled in the art and can stabilize the desired activity of the active ingredient in the formulation, and include, but are not limited to, sodium glutamate, gelatin, SPGA, sugars (such as sorbitol, mannitol, starch, sucrose, lactose, dextran, or glucose), amino acids (such as glutamic acid and glycine), proteins (such as dried whey, albumin, or casein), or their degradation products (such as lactalbumin hydrolysate).

[0138] As used herein, the terms "including," "comprising," "having," "containing," or "involving," and other variations thereof, are inclusive or open-ended and do not exclude other, unrecited elements or method steps.

[0139] The term "effective amount" refers to an amount sufficient to achieve a desired clinical effect in the individual being treated. For example, this may be the amount necessary to alleviate any particular disease symptom in the individual, or to inhibit or reduce the severity of the disease or its complications. Determining such an effective amount is entirely within the capabilities of one skilled in the art. For example, an amount effective for therapeutic use will depend on the severity of the disease being treated, the general condition of the patient's own immune system, the patient's general condition such as age, weight, and sex, the mode of drug administration, and other concurrently administered treatments. The therapeutically effective amount of an ADC may vary according to the following factors: the severity of the disease being treated, the general condition of the patient's own immune system, the patient's general condition such as age, weight, and sex, the mode of drug administration, and other concurrently administered treatments.

[0140] The term "treatment" refers to a method practiced to obtain a beneficial or desired clinical outcome in a subject (human or animal individual) who exhibits disease symptoms or has been diagnosed with a disease ("in need thereof"). For purposes of this invention, beneficial or desired clinical outcomes include, but are not limited to, alleviation of symptoms, reduction in the extent of disease, stabilization of the disease state (i.e., does not worsen), delay or slowing of disease progression, improvement or palliative care of the disease state, and relief of symptoms (whether partial or complete), whether detectable or undetectable. Furthermore, "treatment" can also refer to prolonging survival as compared to expected survival if not receiving treatment.

[0141] As used herein, the term "subject" refers to a mammal, e.g., a primate mammal (e.g., a human). In certain embodiments, the subject (e.g., a human) has a tumor or is at risk of having the above-mentioned disease.

[0142] The terms "cancer" and "tumor" are used interchangeably and refer to a broad class of diseases characterized by the uncontrolled growth of abnormal cells in the body. Uncontrolled cell division can lead to the formation of malignant tumors or cells that can invade adjacent tissues and metastasize to distant sites in the body via the lymphatic system or bloodstream. Cancer includes benign and malignant cancers as well as dormant tumors or micrometastases. Cancer also includes hematologic malignancies.

[0143] The term "hematological malignancies" includes lymphomas, leukemias, myelomas, or lymphatic malignancies, as well as splenic and lymph node neoplasms. Exemplary lymphomas include B-cell lymphomas and T-cell lymphomas. B-cell lymphomas include, for example, Hodgkin's lymphoma. T-cell lymphomas include, for example, cutaneous T-cell lymphoma. Hematological malignancies also include leukemias, such as secondary leukemia or acute lymphoblastic leukemia. Hematological malignancies also include myelomas (e.g., multiple myeloma) and other blood and / or T-cell related cancers.

[0144] The term "alkyl" refers to a group obtained by the loss of one hydrogen atom from a straight or branched chain hydrocarbon group, e.g., "C 1~20 Alkyl," "C 1~10 Alkyl," "C 1~6 Alkyl," "C 1~4 Alkyl" and "C 1~3Illustrative examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, 1,2-dimethylpropyl, and the like.

[0145] The term "alkylene" refers to a group obtained by the loss of two hydrogen atoms from a straight or branched chain hydrocarbon group, e.g., "C 1~20 alkylene," "C 1~10 alkylene," "C 3~10 alkylene," "C 5~8 alkylene," "C 1~6 alkylene," "C 1~4 Alkylene" and "C 1~3 Specific examples include, but are not limited to, methylene, ethylene, 1,3-propylene, 1,4-butylene, 1,5-pentylene, 1,6-hexylene, and the like.

[0146] The term "alkenylene" refers to a divalent group obtained by the loss of two hydrogen atoms from a straight or branched chain hydrocarbon group containing at least one carbon-carbon double bond, e.g., "C 2~20 alkenylene," "C 3~10 alkenylene," "C 5~8 Examples of alkenylene include, but are not limited to, ethenylene, 1-propenylene, 2-propenylene, 1-butenylene, 2-butenylene, 1,3-butadienylene, 1-pentenylene, 2-pentenylene, 3-pentenylene, 1,3-pentadienylene, 1,4-pentadienylene, 1-hexenylene, 2-hexenylene, 3-hexenylene, 1,4-hexadienylene, and the like.

[0147] The term "alkynylene" refers to, for example, "C 2~20 alkynylene," "C 3~10 alkynylene" and "C 5~8 "Alkynylene" refers to a divalent group obtained by the loss of two hydrogen atoms from a straight or branched chain hydrocarbon group containing at least one carbon-carbon triple bond, including "alkynylene." Examples include, but are not limited to, ethynylene, 1-propynylene, 2-propynylene, 1-butynylene, 2-butynylene, 1,3-butadinylene, 1-pentynyl, 2-pentynyl, 3-pentynyl, 1,3-pentadinylene, 1,4-pentadinylene, 1-hexynylene, 2-hexynylene, 3-hexynylene, 1,4-hexadinylene, and the like.

[0148] The term "heteroalicyclic ring" refers to a saturated or partially saturated ring structure containing at least one ring member selected from the group consisting of N, O, and S. Specific examples thereof include, but are not limited to, 5- to 6-membered aliphatic heterocycles, 5- to 6-membered nitrogen-containing aliphatic heterocycles, and 5- to 6-membered oxygen-containing aliphatic heterocycles, such as tetrahydrofuran, pyrrolidine, piperidine, and tetrahydropyran.

[0149] The term "heteroaromatic ring" refers to an aromatic ring structure containing at least one ring member selected from the group consisting of N, O, and S. Specific examples thereof include, but are not limited to, 5- to 6-membered aromatic heterocycles, 5- to 6-membered nitrogen-containing aromatic heterocycles, 5- to 6-membered oxygen-containing heteroaromatic rings, and the like, such as furan, thiophene, pyrrole, thiazole, isothiazole, thiadiazole, oxazole, isoxazole, oxadiazole, imidazole, pyrazole, 1,2,3-triazole, 1,2,4-triazole, 1,2,3-oxadiazole, 1,2,4-oxadiazole, 1,2,5-oxadiazole, 1,3,4-oxadiazole, pyridine, pyrimidine, pyridazine, pyrazine, 1,2,3-triazine, 1,3,5-triazine, and 1,2,4,5-tetrazine.

[0150] The term "aromatic ring system" refers to a monocyclic or polycyclic ring system containing at least one aromatic ring (e.g., a benzene ring) or heteroaromatic ring (e.g., a pyrimidine ring), where two or more aromatic and / or heteroaromatic rings may form fused rings or be connected by a single bond (e.g., dipyrimidinylphenyl), and the aromatic ring system may be divalent or polyvalent (e.g., trivalent or tetravalent), e.g., a 5- to 20-membered aromatic ring system.

[0151] As used herein, "Cit" is an abbreviation for citrulline.

[0152] As used herein, the terms "about" or "approximately," when used in conjunction with a numerical variable, generally mean that the value of the variable is within experimental error (e.g., within a 95% confidence interval of the mean) or within ±10%.

[0153] It should be noted that if there is a discrepancy between a depicted structure and the name of that structure, the depicted structure takes precedence.

[0154] antibody-drug conjugates In one aspect, the present application provides a compound of formula Ab-[MLED] x An antibody-drug conjugate having the structure shown in Ab is an antibody or antigen-binding fragment thereof that specifically binds to human tyrosine kinase 7 (PTK7); M is a binding site linked to an antibody or antigen-binding fragment thereof; L is a linker between M and E, E is a structural fragment connecting L and D, D is a cytotoxic drug fragment; An antibody-drug conjugate wherein x is selected from 1 to 10.

[0155] In some embodiments, the antibody or antigen-binding fragment thereof (1) A heavy chain variable region (VH) and / or light chain variable region (VL) of the following, wherein the CDRs are defined according to the Chothia numbering system: (1a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 11 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 12 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof; or (1b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 27 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 28 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof, The variants of either (1a) and (1b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with respect to the sequence from which the variant is derived, or have one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, preferably conservative substitutions, in a heavy chain variable region (VH) and / or a light chain variable region (VL); or (2) A heavy chain variable region (VH) and / or a light chain variable region (VL) of the following, wherein the CDRs are defined according to the Kabat numbering system: (2a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 17 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 18 or 19 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof; or (2b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 33 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 34 or 35 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof; The variants of either (2a) and (2b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with respect to the sequence from which the variant is derived, or have one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, preferably conservative substitutions, in a heavy chain variable region (VH) and / or a light chain variable region (VL); or (3) A heavy chain variable region (VH) and / or light chain variable region (VL) of the following, wherein the CDRs are defined according to the IMGT numbering system: (3a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 20 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 21 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 22 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 23 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 24 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof; or (3b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 36 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 37 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 38 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 39 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 40 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof; The variants of either (3a) and (3b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with respect to the sequence from which the variant is derived, or have one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, and the substitutions are preferably conservative substitutions. or (4) A heavy chain variable region (VH) and / or light chain variable region (VL) of the following, wherein the CDRs are defined according to the AbM numbering system: (4a) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 25 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 26 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof, or (4b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 41 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 42 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof; The variants of either (4a) or (4b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence from which the variant is derived, or have undergone one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, and preferably, the substitutions are conservative substitutions.

[0156] In some embodiments, the antibody or antigen-binding fragment thereof (1a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 11 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 12 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof; or (1b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 27 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 28 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof, The variants of either (1a) and (1b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with respect to the sequence from which the variant is derived, or have one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, preferably conservative substitutions, in a heavy chain variable region (VH) and / or a light chain variable region (VL); or (2a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 17 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 18 or 19 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof; or (2b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 33 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 34 or 35 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof; The variants of either (2a) and (2b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with respect to the sequence from which the variant is derived, or have one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, preferably conservative substitutions, in a heavy chain variable region (VH) and / or a light chain variable region (VL); or (3a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 20 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 21 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 22 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 23 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 24 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof; or (3b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 36 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 37 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 38 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 39 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 40 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof; The variants of either (3a) and (3b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with respect to the sequence from which the variant is derived, or have one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, and the substitutions are preferably conservative substitutions. or (4a) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 25 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 26 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof, or (4b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 41 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 42 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof; The variants of either (4a) and (4b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence from which the variant is derived, or have one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, and preferably the substitutions are conservative substitutions.

[0157] In some embodiments, the antibody or antigen-binding fragment thereof (1a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 11, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 12, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13; and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16; or (1b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 27, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 28, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29; and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32; or (2a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 17, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 18 or 19, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13; and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16; or (2b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 33, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 34 or 35, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29; and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32; or (3a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 20, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 21, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 22; and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 23, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 24, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16; or (3b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 36, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 37, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 38; and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 39, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 40, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32; or (4a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 25, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 26, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13; and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16; or (4b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 41, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 42, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29; and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32;

[0158] In some embodiments, the antibody or antigen-binding fragment thereof comprises (1a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 11, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 12, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16.

[0159] In some embodiments, the antibody or antigen-binding fragment thereof comprises (1b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 27, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 28, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32.

[0160] In some embodiments, the antibody or antigen-binding fragment thereof comprises (2a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 17, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 18 or 19, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16.

[0161] In some embodiments, the antibody or antigen-binding fragment thereof comprises (2b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 33, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 34 or 35, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32.

[0162] In some embodiments, the antibody or antigen-binding fragment thereof comprises (3a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 20, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 21, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 22, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 23, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 24, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16.

[0163] In some embodiments, the antibody or antigen-binding fragment thereof comprises (3b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 36, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 37, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 38, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 39, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 40, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32.

[0164] In some embodiments, the antibody or antigen-binding fragment thereof comprises (4a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 25, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 26, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16.

[0165] In some embodiments, the antibody or antigen-binding fragment thereof comprises (4b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 41, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 42, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32.

[0166] In some embodiments, the antibody or antigen-binding fragment thereof (a) a VH having the amino acid sequence set forth in SEQ ID NO: 1 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 2 or a variant thereof; (b) a VH having the amino acid sequence set forth in SEQ ID NO: 3 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 4 or a variant thereof; (c) a VH having the amino acid sequence set forth in SEQ ID NO: 5 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 6 or a variant thereof; (d) a VH having the amino acid sequence set forth in SEQ ID NO: 7 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 8 or a variant thereof; or (e) a VH having the amino acid sequence set forth in SEQ ID NO: 9 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 10 or a variant thereof; A variant may have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence from which it is derived, or may have one or several amino acid substitutions, deletions, or additions (e.g., 1, 2, 3, 4, or 5 amino acid substitutions, deletions, or additions) with respect to the sequence from which it is derived, preferably wherein the substitutions are conservative substitutions.

[0167] In some embodiments, the antibody or antigen-binding fragment thereof (a) a VH having the amino acid sequence set forth in SEQ ID NO: 1, and a VL having the amino acid sequence set forth in SEQ ID NO: 2; (b) a VH having the amino acid sequence set forth in SEQ ID NO: 3, and a VL having the amino acid sequence set forth in SEQ ID NO: 4; (c) a VH having the amino acid sequence set forth in SEQ ID NO: 5, and a VL having the amino acid sequence set forth in SEQ ID NO: 6; (d) a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a VL having the amino acid sequence set forth in SEQ ID NO: 8; or (e) VH having the amino acid sequence set forth in SEQ ID NO: 9, and VL having the amino acid sequence set forth in SEQ ID NO: 10.

[0168] In some embodiments, the antibody or antigen-binding fragment thereof comprises (a) a VH having the amino acid sequence set forth in SEQ ID NO:1, and / or a VL having the amino acid sequence set forth in SEQ ID NO:2.

[0169] In some embodiments, the antibody or antigen-binding fragment thereof comprises (b) a VH having the amino acid sequence set forth in SEQ ID NO:3, and / or a VL having the amino acid sequence set forth in SEQ ID NO:4.

[0170] In some embodiments, the antibody or antigen-binding fragment thereof comprises (c) a VH having the amino acid sequence set forth in SEQ ID NO:5, and / or a VL having the amino acid sequence set forth in SEQ ID NO:6.

[0171] In some embodiments, the antibody or antigen-binding fragment thereof comprises (d) a VH having the amino acid sequence set forth in SEQ ID NO:7, and / or a VL having the amino acid sequence set forth in SEQ ID NO:8.

[0172] In some embodiments, the antibody or antigen-binding fragment thereof comprises (e) a VH having the amino acid sequence set forth in SEQ ID NO:9, and / or a VL having the amino acid sequence set forth in SEQ ID NO:10.

[0173] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) comprising CDR-H1, CDR-H2, and CDR-H3, the amino acid sequences of which are set forth in VH, respectively, having the amino acid sequence of SEQ ID NO: 1, and a light chain variable region (VL) comprising CDR-L1, CDR-L2, and CDR-L3, the amino acid sequences of which are set forth in VL, respectively, having the amino acid sequence of SEQ ID NO: 2.

[0174] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) comprising CDR-H1, CDR-H2, and CDR-H3, the VH of which comprises the amino acid sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, set forth in SEQ ID NO: 3, and a light chain variable region (VL) comprising CDR-L1, CDR-L2, and CDR-L3, the VL of which comprises the amino acid sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, set forth in SEQ ID NO: 4.

[0175] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) comprising CDR-H1, CDR-H2, and CDR-H3, the VH of which comprises the amino acid sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, set forth in SEQ ID NO: 5, and a light chain variable region (VL) comprising CDR-L1, CDR-L2, and CDR-L3, the VL of which comprises the amino acid sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, set forth in SEQ ID NO: 6.

[0176] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) comprising CDR-H1, CDR-H2, and CDR-H3, the amino acid sequences of which are set forth in VH, respectively, having the amino acid sequence of SEQ ID NO: 7, and a light chain variable region (VL) comprising CDR-L1, CDR-L2, and CDR-L3, the amino acid sequences of which are set forth in VL, respectively, having the amino acid sequence of SEQ ID NO: 8.

[0177] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) comprising CDR-H1, CDR-H2, and CDR-H3, the VH of which comprises the amino acid sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, set forth in SEQ ID NO: 9, and a light chain variable region (VL) comprising CDR-L1, CDR-L2, and CDR-L3, the VL of which comprises the amino acid sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, set forth in SEQ ID NO: 10.

[0178] In some embodiments, the antibody or antigen-binding fragment thereof (a) a human immunoglobulin heavy chain constant region (CH) or a variant thereof, wherein the variant has one or more amino acid substitutions, deletions, or additions (e.g., up to 20, up to 15, up to 10, or up to 5 amino acid substitutions, deletions, or additions, e.g., 1, 2, 3, 4, or 5 amino acid substitutions, deletions, or additions) compared to the wild-type sequence from which it is derived; and (b) A human immunoglobulin light chain constant region (CL) or variant thereof, wherein the variant further comprises one or more amino acid substitutions, deletions, or additions (e.g., up to 20, up to 15, up to 10, or up to 5 amino acid substitutions, deletions, or additions, e.g., 1, 2, 3, 4, or 5 amino acid substitutions, deletions, or additions) compared to the wild-type sequence from which it is derived.

[0179] In some embodiments, the heavy chain constant region is an IgG heavy chain constant region, e.g., an IgG1, IgG2, IgG3, or IgG4 heavy chain constant region, e.g., a human IgG1 heavy chain constant region or a human IgG4 heavy chain constant region. In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain constant region (CH) set forth as SEQ ID NO: 43 or a variant thereof, wherein the variant has up to 20 amino acid conservative substitutions (e.g., up to 15, up to 10, or up to 5 amino acid conservative substitutions, e.g., 1, 2, 3, 4, or 5 amino acid conservative substitutions) compared to SEQ ID NO: 43.

[0180] In some embodiments, the light chain constant region comprises a kappa light chain constant region. In some embodiments, the antibody, or antigen-binding region thereof, comprises a light chain constant region (CL) set forth as SEQ ID NO: 44, or a variant thereof, wherein the variant has up to 20 amino acid conservative substitutions (e.g., up to 15, up to 10, or up to 5 amino acid conservative substitutions, e.g., 1, 2, 3, 4, or 5 amino acid conservative substitutions) compared to SEQ ID NO: 44.

[0181] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain constant region (CH) set forth as SEQ ID NO: 43 or 45 and a light chain constant region (CL) set forth as SEQ ID NO: 44.

[0182] In some embodiments, the antibody or antigen-binding fragment thereof (1) A heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43, and a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44; (2) a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44; (3) a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43, and a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44; (4) A heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43, and a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 8 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44, or (5) A heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43, and a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 10 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44.

[0183] In some embodiments, the antibody or antigen-binding fragment thereof comprises: (1) a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43, and a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44.

[0184] In some embodiments, the antibody or antigen-binding fragment thereof comprises (2) a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44.

[0185] In some embodiments, the antibody or antigen-binding fragment thereof comprises (3) a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO:5 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO:43, and a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO:6 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO:44.

[0186] In some embodiments, the antibody or antigen-binding fragment thereof comprises (4) a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO:7 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO:43, and a light chain comprising a VL having the amino acid sequence set forth as SEQ ID NO:8 and a light chain constant region (CL) having the amino acid sequence set forth as SEQ ID NO:44.

[0187] In some embodiments, the antibody or antigen-binding fragment thereof comprises (5) a heavy chain comprising a VH set forth as SEQ ID NO: 9 and a heavy chain constant region (CH) set forth as SEQ ID NO: 43, and a light chain comprising a VL set forth as SEQ ID NO: 10 and a light chain constant region (CL) set forth as SEQ ID NO: 44.

[0188] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO:46, and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO:47.

[0189] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO:52, and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO:47.

[0190] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO:53, and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO:47.

[0191] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO:54, and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO:47.

[0192] In certain embodiments of the antibodies or antibody-drug conjugates disclosed herein, the heavy chain constant domain may include a C-terminal lysine or may lack either the C-terminal lysine or the C-terminal glycine-lysine dipeptide. In certain embodiments of the antibodies or antibody-drug conjugates thereof, the N-terminal amino acid of the antibody variable domain may undergo cyclization to pyroglutamate.

[0193] In certain embodiments of the antibodies or antibody-drug conjugates disclosed herein, the N-terminal amino acid of the antibody variable domain may undergo cyclization to pyroglutamic acid.

[0194] In certain embodiments, the antibodies or antigen-binding fragments disclosed herein include antibodies or antigen-binding fragments that specifically bind to an antigen and may include post-translational modifications thereof (e.g., C-terminal lysine clipping in the heavy chain, conversion of glutamine or glutamic acid to pyroglutamate or pyroglutamic acid) that may occur when recombinantly expressed in a host cell (e.g., a CHO cell) or during purification / storage.

[0195] Thus, a composition can comprise a population of antibody-drug conjugate species, each of which can independently comprise a C-terminal lysine, lack a C-terminal lysine, lack a C-terminal glycine-lysine, and / or comprise an N-terminal glutamine or glutamic acid or cyclization of the N-terminal amino acid to pyroglutamate.

[0196] In certain embodiments, a composition may comprise a population of antibody-drug conjugate species, each of which may independently comprise a C-terminal lysine, lack a C-terminal lysine, lack a C-terminal glycine-lysine, and / or comprise an N-terminal glutamine or glutamic acid or cyclization of the N-terminal amino acid to pyroglutamic acid.

[0197] Accordingly, in certain embodiments, the present invention further provides a composition comprising an ADC disclosed herein, wherein the predominant ADC species in the composition comprises: (i) an antibody whose heavy chain C-terminus lacks a lysine residue; (ii) an antibody whose heavy chain N-terminus is glutamine, glutamic acid, pyroglutamate, or pyroglutamic acid; or (iii) an antibody whose heavy chain C-terminus lacks a lysine residue and whose heavy chain N-terminus is glutamine, glutamic acid, pyroglutamate, or pyroglutamic acid.

[0198] Accordingly, in certain embodiments, the invention further provides a composition comprising an ADC disclosed herein, wherein the predominant ADC species in the composition comprises (i) an antibody whose heavy chain C-terminus lacks a lysine residue and whose heavy chain N-terminus is pyroglutamate or pyroglutamic acid.

[0199] In certain embodiments, the antibody or antigen-binding fragment thereof according to the invention comprises a heavy chain comprising a VH having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, and 9.

[0200] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the invention comprises a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO:1.

[0201] In a specific embodiment, an antibody or antigen-binding fragment thereof according to the invention comprises a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO:3.

[0202] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the invention comprises a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO:5.

[0203] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the invention comprises a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO:7.

[0204] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the invention comprises a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO:9.

[0205] In certain embodiments, the antibody or antigen-binding fragment thereof according to the invention comprises a heavy chain comprising a VH having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, and 9, wherein the N-terminal glutamine or glutamic acid of the antibody heavy chain variable region has been cyclized to pyroglutamate.

[0206] In certain embodiments, the antibody or antigen-binding fragment thereof according to the invention comprises a heavy chain comprising a VH having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, and 9, wherein the N-terminal glutamine or glutamic acid of the antibody heavy chain variable region has been cyclized to pyroglutamic acid.

[0207] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the invention comprises a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1, wherein the N-terminal glutamic acid of the antibody heavy chain variable region has been cyclized to pyroglutamate.

[0208] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the present invention comprises a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1, wherein the N-terminal glutamic acid of the antibody heavy chain variable region is cyclized to pyroglutamic acid.

[0209] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the invention comprises a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3, wherein the N-terminal glutamic acid of the antibody heavy chain variable region has been cyclized to pyroglutamate.

[0210] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the present invention comprises a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3, wherein the N-terminal glutamic acid of the antibody heavy chain variable region is cyclized to pyroglutamic acid.

[0211] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the invention comprises a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5, wherein the N-terminal glutamine of the antibody heavy chain variable region has been cyclized to pyroglutamate.

[0212] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the invention comprises a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5, wherein the N-terminal glutamic acid of the antibody heavy chain variable region is cyclized to pyroglutamic acid.

[0213] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the invention comprises a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7, wherein the N-terminal glutamine of the antibody heavy chain variable region has been cyclized to pyroglutamate.

[0214] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the present invention comprises a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7, wherein the N-terminal glutamic acid of the antibody heavy chain variable region is cyclized to pyroglutamic acid.

[0215] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the invention comprises a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9, wherein the N-terminal glutamine of the antibody heavy chain variable region has been cyclized to pyroglutamate.

[0216] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the invention comprises a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9, wherein the N-terminal glutamic acid of the antibody heavy chain variable region is cyclized to pyroglutamic acid.

[0217] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the invention comprises a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 8, wherein the N-terminal glutamic acid of the antibody light chain variable region has been cyclized to pyroglutamate.

[0218] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the invention comprises a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 8, wherein the N-terminal glutamic acid of the antibody light chain variable region is cyclized to pyroglutamic acid.

[0219] In certain embodiments, an antibody or antigen-binding fragment thereof according to the invention comprises a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7, and / or a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 8, wherein the N-terminal glutamine of SEQ ID NO: 7 and / or the N-terminal glutamic acid of SEQ ID NO: 8 has been cyclized to pyroglutamate.

[0220] In certain embodiments, an antibody or antigen-binding fragment thereof according to the invention comprises a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and / or a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 8, wherein the N-terminal glutamine of SEQ ID NO: 7 and / or the N-terminal glutamic acid of SEQ ID NO: 8 has been cyclized to pyroglutamic acid.

[0221] In certain embodiments, the antibody or antigen-binding fragment (a) a heavy chain consisting of the sequence set forth as SEQ ID NO: 52 and a light chain consisting of the sequence set forth as SEQ ID NO: 47; (b) a heavy chain consisting of the sequence set forth as SEQ ID NO: 53 and a light chain consisting of the sequence set forth as SEQ ID NO: 47; or (c) a heavy chain consisting of the sequence shown as SEQ ID NO: 54 and a light chain consisting of the sequence shown as SEQ ID NO: 47.

[0222] In certain embodiments, the antibody or antigen-binding fragment thereof according to the invention can be obtained by expressing nucleic acid molecules encoding the heavy and light chains of the antibody or antigen-binding fragment thereof in a host cell, wherein the antibody or antigen-binding fragment thereof comprises: (a) a VH having the amino acid sequence set forth in SEQ ID NO: 1 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 2 or a variant thereof; (b) a VH having the amino acid sequence set forth in SEQ ID NO: 3 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 4 or a variant thereof; (c) a VH having the amino acid sequence set forth in SEQ ID NO: 5 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 6 or a variant thereof; (d) a VH having the amino acid sequence set forth in SEQ ID NO: 7 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 8 or a variant thereof; or (e) a VH having the amino acid sequence set forth in SEQ ID NO: 9 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 10 or a variant thereof; The variant has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity compared to the sequence from which the variant is derived, or has one or several amino acid substitutions, deletions, or additions (e.g., 1, 2, 3, 4, or 5 amino acid substitutions, deletions, or additions) compared to the sequence from which the variant is derived.

[0223] In certain embodiments, the substitution is a conservative substitution. In certain embodiments, the antibody or antigen-binding fragment thereof according to the invention can be obtained by expressing nucleic acid molecules encoding the heavy and light chains of the antibody or antigen-binding fragment thereof in a host cell, and the antibody or antigen-binding fragment thereof comprises: (a) a VH having the amino acid sequence set forth in SEQ ID NO: 1, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 2; (b) a VH having the amino acid sequence set forth in SEQ ID NO: 3, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 4; (c) a VH having the amino acid sequence set forth in SEQ ID NO: 5, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 6; (d) a VH having the amino acid sequence set forth in SEQ ID NO: 7, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 8, or (e) VH having the amino acid sequence set forth in SEQ ID NO: 9, and / or VL having the amino acid sequence set forth in SEQ ID NO: 10.

[0224] In certain embodiments, the antibody or antigen-binding fragment thereof can be obtained by expressing in a host cell nucleic acid molecules encoding the heavy and light chains of the antibody or antigen-binding fragment thereof, and the antibody or antigen-binding fragment thereof can be (1) a heavy chain comprising a VH sequence set forth as SEQ ID NO: 1 and a heavy chain constant region (CH) set forth as SEQ ID NO: 43, and a light chain comprising a VL sequence set forth as SEQ ID NO: 2 and a light chain constant region (CL) set forth as SEQ ID NO: 44; (2) a heavy chain comprising a VH sequence set forth as SEQ ID NO: 3 and a heavy chain constant region (CH) set forth as SEQ ID NO: 43 or 45, and a light chain comprising a VL sequence set forth as SEQ ID NO: 4 and a light chain constant region (CL) set forth as SEQ ID NO: 44; (3) a heavy chain comprising a VH sequence set forth as SEQ ID NO: 5 and a heavy chain constant region (CH) set forth as SEQ ID NO: 43, and a light chain comprising a VL sequence set forth as SEQ ID NO: 6 and a light chain constant region (CL) set forth as SEQ ID NO: 44; (4) a heavy chain comprising a VH sequence set forth as SEQ ID NO: 7 and a heavy chain constant region (CH) set forth as SEQ ID NO: 43, and a light chain comprising a VL sequence set forth as SEQ ID NO: 8 and a light chain constant region (CL) set forth as SEQ ID NO: 44; (5) a heavy chain comprising a VH sequence set forth as SEQ ID NO: 9 and a heavy chain constant region (CH) set forth as SEQ ID NO: 43, and a light chain comprising a VL sequence set forth as SEQ ID NO: 10 and a light chain constant region (CL) set forth as SEQ ID NO: 44; (6) a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46, and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47; (7) VH having the amino acid sequence set forth in SEQ ID NO: 1, and VL having the amino acid sequence set forth in SEQ ID NO: 2; (8) VH having the amino acid sequence set forth in SEQ ID NO: 3, and VL having the amino acid sequence set forth in SEQ ID NO: 4; (9) VH having the amino acid sequence set forth in SEQ ID NO: 5, and VL having the amino acid sequence set forth in SEQ ID NO: 6; (10) VH having the amino acid sequence set forth in SEQ ID NO: 7 and VL having the amino acid sequence set forth in SEQ ID NO: 8, or (11) A VH having the amino acid sequence set forth in SEQ ID NO: 9 and a VL having the amino acid sequence set forth in SEQ ID NO: 10.

[0225] In certain embodiments, the antibody or antigen-binding fragment thereof according to the present invention can be obtained by expressing in a host cell nucleic acid molecules encoding the heavy and light chains of the antibody or antigen-binding fragment thereof, and the antibody or antigen-binding fragment thereof comprises (a) a VH having the amino acid sequence set forth in SEQ ID NO: 1 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 2 or a variant thereof.

[0226] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the present invention can be obtained by expressing in a host cell nucleic acid molecules encoding the heavy and light chains of the antibody or antigen-binding fragment thereof, and the antibody or antigen-binding fragment thereof comprises (b) a VH having the amino acid sequence set forth in SEQ ID NO: 3 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 4 or a variant thereof.

[0227] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the present invention can be obtained by expressing in a host cell nucleic acid molecules encoding the heavy and light chains of the antibody or antigen-binding fragment thereof, and the antibody or antigen-binding fragment thereof comprises (c) a VH having the amino acid sequence set forth in SEQ ID NO: 5 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 6 or a variant thereof.

[0228] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the present invention can be obtained by expressing in a host cell nucleic acid molecules encoding the heavy and light chains of the antibody or antigen-binding fragment thereof, and the antibody or antigen-binding fragment thereof comprises (d) a VH having the amino acid sequence set forth in SEQ ID NO: 7 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 8 or a variant thereof.

[0229] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the present invention can be obtained by expressing in a host cell nucleic acid molecules encoding the heavy and light chains of the antibody or antigen-binding fragment thereof, and the antibody or antigen-binding fragment thereof comprises (e) a VH having the amino acid sequence set forth in SEQ ID NO: 9 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 10 or a variant thereof.

[0230] In certain antibody embodiments, the heavy chain constant domains disclosed herein may include a C-terminal lysine or may lack either a C-terminal lysine or a C-terminal glycine-lysine dipeptide. In some antibody or antigen-binding fragment thereof embodiments, the N-terminal amino acid of the antibody or antigen-binding fragment thereof variable domain may undergo cyclization to pyroglutamate.

[0231] In some embodiments of the antibody or antigen-binding fragment thereof, the N-terminal amino acid of the antibody or antigen-binding fragment thereof variable domain may undergo cyclization to pyroglutamic acid.

[0232] Thus, in compositions comprising particular antibodies disclosed herein, the composition may comprise a population of antibody species, each of which may independently comprise a C-terminal lysine, lack a C-terminal lysine, lack a C-terminal glycine-lysine, and / or comprise an N-terminal glutamine or glutamic acid or cyclization of the N-terminal amino acid to pyroglutamate.

[0233] In certain embodiments, in compositions comprising particular antibodies disclosed herein, the composition may comprise a population of antibody species, each of which may independently comprise a C-terminal lysine, may lack a C-terminal lysine, may lack a C-terminal glycine-lysine, and / or may comprise an N-terminal glutamine or glutamic acid or cyclization of the N-terminal amino acid to pyroglutamic acid.

[0234] In the antibody-drug conjugate, the cytotoxic drug may be linked to the antibody or antigen-binding fragment via a linker (e.g., the "MLE" fragment shown in this application).

[0235] In some embodiments, M is [ka] wherein ring A is a 5- to 6-membered aliphatic heterocycle or a 5- to 20-membered aromatic ring system, and the aliphatic heterocycle and aromatic ring system are substituted with oxo (=O), halogen, cyano, amino, carboxyl, mercapto, and C 1~6alkyl; and M is a single bond, C 1~20 Alkylene, C 2~20 Alkenylene, and C 2~20 alkynylene.

[0236] In some embodiments, M is [ka] wherein ring A is a 5-membered aliphatic heterocycle, a 6-membered aromatic heterocycle, or a polycyclic ring formed by connecting two or more 6-membered aromatic heterocycles and a benzene ring with a single bond, and the aliphatic heterocycle is selected from the group consisting of oxo (=O), halogen, and C 1~4 alkyl, and M is a single bond, C 3~10 Alkylene, C 3~10 Alkenylene, and C 3~10 alkynylene.

[0237] In some embodiments, M is [ka] and ring A is [ka] M1 is selected from a single bond, C 5~8 Alkylene, C 5~8 Alkenylene, and C 5~8 alkynylene.

[0238] In some embodiments, M has the following structure: [ka] is selected from.

[0239] In some embodiments, M is selected from the following structures: [ka]

[0240] In some embodiments, L is C 1~6 Alkylene, -N(R')-, carbonyl, -O-, Val, Cit, Phe, Lys, Lys(COCH2CH2(OCH2CH2) s OCH3), D-Val, Leu, Gly, Ala, Asn, Val-Cit, Val-Ala, Val-Lys, Val-Lys(Ac), Phe-Lys, Phe-Lys(Ac), D-Val-Leu-Lys, Gly-Gly-Arg, Ala-Ala-Asn, Ala-Ala-Ala, Val-Lys-Ala, Val-Lys-Gly, Gly-Gly-Gly, Gly-Gly-Phe-Gly (SEQ ID NO: 48), Gly-Phe-Leu-Gly (SEQ ID NO: 49), Gly-Gly-Val-Ala (SEQ ID NO: 50), Gly-Gly-Gly-Gly-Gly (SEQ ID NO: 51), [ka] and R' is selected from the group consisting of hydrogen, C 1~6 alkyl, or alkyl containing -(CH2CH2O)r-, where r is selected from an integer of 1 to 10, and s is selected from an integer of 1 to 20.

[0241] In some embodiments, L is C 1~6 Alkylene, -NH-, Val, Cit, Phe, Lys, Lys(COCH2CH2(OCH2CH2) s OCH3), Gly, Val-Cit, Gly-Gly-Phe-Gly (SEQ ID NO: 48), [ka] and s is selected from an integer of 1 to 20.

[0242] In some embodiments, L has the following structure: [ka] is selected from s is selected from integers of 1 to 20.

[0243] In some embodiments, L has the following structure: [ka] is selected from.

[0244] In some embodiments, L has the following structure: [ka] is selected from.

[0245] In some embodiments, L has the following structure: [ka] is selected from.

[0246] In some embodiments, E is a single bond, —NH—CH 2 —, —NH—CH 2 —O—CH 2 —CO—, [ka] is.

[0247] In some embodiments, E is a single bond, —NH—CH 2 —, —NH—CH 2 —O—CH 2 —CO—, [ka] is.

[0248] In some embodiments, E is -NH-CH2-, [ka] is.

[0249] In some embodiments, E is -NH-CH2- or [ka] is.

[0250] In some embodiments, M has the following structure: [ka] is selected from L has the following structure: [ka] is selected from E is -NH-CH2, [ka] is.

[0251] In some embodiments, [ka] is selected from the following structures: [ka] [ka] [ka]

[0252] In some embodiments, [ka] is selected from the following structures: [ka] In some embodiments, [ka] is selected from the following structures: [ka] , [ka] , [ka] , [ka] , [ka] , [ka]

[0253] In some embodiments, the cytotoxic drug is selected from a tubulin inhibitor, a DNA intercalator, a DNA topoisomerase inhibitor, and an RNA polymerase inhibitor. In some embodiments, the tubulin inhibitor is an auristatin compound or a maytansinoid compound. In some embodiments, the DNA intercalator is a pyrrolobenzodiazepine (PBD). In some embodiments, the DNA topoisomerase inhibitor is a topoisomerase I inhibitor (e.g., camptothecin, hydroxycamptothecin, 9-aminocamptothecin, SN-38, irinotecan, topotecan, belotecan, or rubitecan) or a topoisomerase II inhibitor (e.g., doxorubicin, PNU-159682, duocarmycin, daunorubicin, mitoxantrone, podophyllotoxin, or etoposide). In some embodiments, the RNA polymerase inhibitor is α-amanitin or a pharmaceutically acceptable salt, ester, or analog thereof.

[0254] Cytotoxic drugs disclosed in this application generally contain hydroxyl (-OH), carboxyl (-COOH), sulfhydryl (-SH), primary amino (-NH), secondary amine groups (-NR A H), or a tertiary amine group (-NR B R C ), and R A , R B , and R C represents only non-hydrogen substituents on the N, and the cytotoxic drug can be linked to the linker in the conjugate via these functional groups.

[0255] In some embodiments, the cytotoxic agent is linked to E in the antibody-drug conjugate via -OH, -SH, a primary amine group, a secondary amine group, or a tertiary amine group.

[0256] In some embodiments, the cytotoxic agent is [ka] is.

[0257] In some embodiments, the cytotoxic agent is selected from Formulas I and II: [ka] During the ceremony, R1 and R2 each independently represent C 1~6 selected from alkyl and halogen; R3 is selected from H and —CO—CH2OH; R4 and R5 are each independently selected from H, halogen, and hydroxyl, or R4 and R5 are joined to associated carbon atoms to form a 5- to 6-membered oxygen-containing heterocycle; R6 is hydrogen and -C 1~4 Alkylene-NR a R b is selected from R7 is hydrogen, C 1~6 Alkyl, and -C 1~4 Alkylene-NRa R b is selected from In each occurrence, R a and R b are each independently H, C 1~6 Alkyl, -SO2-C 1~6 Alkyl, and -CO-C 1~6 alkyl.

[0258] In some embodiments, the cytotoxic agent has the following structure: [ka] is selected from.

[0259] In some embodiments, the cytotoxic agent has the following structure: [ka] is selected from.

[0260] The corresponding fragment of the cytotoxic drug obtained after the cytotoxic drug is linked to the linker has the formula Ab-[MLED] according to the present application. x In some embodiments, D is a monovalent structure obtained by loss of one H from an -OH, -NH2, or secondary amine group on the cytotoxic drug.

[0261] In some embodiments, D has the following structure: [ka] is selected from.

[0262] In some embodiments, the antibody-drug conjugate is selected from ADC A-01 to ADC A-26, ADC B-01 to ADC B-06, and ADC C-01 shown below. [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] the HA in each antibody-drug conjugate represents an antibody or antigen-binding fragment of a VH set forth in SEQ ID NO: 1, 3, 5, 7, or 9 and a VL set forth in SEQ ID NO: 2, 4, 6, 8, or 10, e.g., an antibody or antigen-binding fragment of a VH set forth in SEQ ID NO: 3 and a CH set forth in SEQ ID NO: 43 or 45, a VL set forth in SEQ ID NO: 4, and a CL set forth in SEQ ID NO: 44; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, wherein: HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0263] In some embodiments, the antibody-drug conjugate comprises: [ka] [ka] is selected from. The HA in each antibody-drug conjugate was (1) An antibody or antigen-binding fragment thereof comprising: a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45; a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (2) An antibody or antigen-binding fragment thereof comprising: a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45; and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a CL having the amino acid sequence set forth in SEQ ID NO: 44. (3) An antibody or antigen-binding fragment thereof comprising: a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45; and a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a CL having the amino acid sequence set forth in SEQ ID NO: 44. (4) An antibody or antigen-binding fragment thereof comprising: a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45; and a VL having the amino acid sequence set forth in SEQ ID NO: 8 and a CL having the amino acid sequence set forth in SEQ ID NO: 44. (5) An antibody or antigen-binding fragment thereof comprising: a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45; and a VL having the amino acid sequence set forth in SEQ ID NO: 10 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate. x is 3 to 9.

[0264] In some embodiments, the antibody-drug conjugate comprises: [ka] [ka] During the ceremony, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate; x is 2 to 8, or 3 to 4, or 7 to 8, and the HA in each antibody-drug conjugate is (1) A heavy chain variable region (VH) and / or light chain variable region (VL) of the following, wherein the CDRs are defined according to the Chothia numbering system: (1a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 11 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 12 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof; or (1b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 27 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 28 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof, The variants of either (1a) and (1b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with respect to the sequence from which the variant is derived, or have one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, preferably conservative substitutions, in a heavy chain variable region (VH) and / or a light chain variable region (VL); or (2) A heavy chain variable region (VH) and / or a light chain variable region (VL) of the following, wherein the CDRs are defined according to the Kabat numbering system: (2a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 17 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 18 or 19 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof; or (2b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 33 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 34 or 35 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof; The variants of either (2a) and (2b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with respect to the sequence from which the variant is derived, or have one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, preferably conservative substitutions, in a heavy chain variable region (VH) and / or a light chain variable region (VL); or (3) A heavy chain variable region (VH) and / or light chain variable region (VL) of the following, wherein the CDRs are defined according to the IMGT numbering system: (3a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 20 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 21 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 22 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 23 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 24 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof; or (3b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 36 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 37 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 38 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 39 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 40 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof; The variants of either (3a) and (3b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with respect to the sequence from which the variant is derived, or have one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, and the substitutions are preferably conservative substitutions. or (4) A heavy chain variable region (VH) and / or light chain variable region (VL) of the following, wherein the CDRs are defined according to the AbM numbering system: (4a) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 25 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 26 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof, or (4b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 41 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 42 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof; The variants of either (4a) or (4b) are selected from antibodies or antigen-binding fragments comprising a heavy chain variable region (VH) and / or a light chain variable region (VL) that have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence from which the variant is derived, or that have undergone one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, and preferably the substitutions are conservative substitutions.

[0265] In a further embodiment, the HA in each antibody-drug conjugate is (a) a VH having the amino acid sequence set forth in SEQ ID NO: 1 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 2 or a variant thereof; (b) a VH having the amino acid sequence set forth in SEQ ID NO: 3 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 4 or a variant thereof; (c) a VH having the amino acid sequence set forth in SEQ ID NO: 5 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 6 or a variant thereof; (d) a VH having the amino acid sequence set forth in SEQ ID NO: 7 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 8 or a variant thereof; and (e) a VH having the amino acid sequence set forth in SEQ ID NO: 9 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 10 or a variant thereof; The variants according to any of items (a), (b), (c), (d), and (e) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% amino acid sequence identity compared to the sequence from which the variant is derived, or the variants are selected from antibodies or antigen-binding fragments having one or several amino acid substitutions, deletions, or additions (e.g., 1, 2, 3, 4, or 5 amino acid substitutions, deletions, or additions) compared to the sequence from which the variant is derived, preferably wherein the substitutions are conservative substitutions.

[0266] In a further embodiment, the HA in each antibody-drug conjugate is (a) a VH having the amino acid sequence set forth in SEQ ID NO: 1, and a VL having the amino acid sequence set forth in SEQ ID NO: 2; (b) a VH having the amino acid sequence set forth in SEQ ID NO: 3, and a VL having the amino acid sequence set forth in SEQ ID NO: 4; (c) a VH having the amino acid sequence set forth in SEQ ID NO: 5, and a VL having the amino acid sequence set forth in SEQ ID NO: 6; (a) a VH having the amino acid sequence set forth in SEQ ID NO: 7, and a VL having the amino acid sequence set forth in SEQ ID NO: 8; (a) an antibody or antigen-binding fragment comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a VL having the amino acid sequence set forth in SEQ ID NO: 10;

[0267] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, and 9.

[0268] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO:1.

[0269] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO:3.

[0270] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO:5.

[0271] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO:7.

[0272] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO:9.

[0273] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, and 9, and wherein the N-terminal glutamine or glutamic acid of the antibody variable region has been cyclized to pyroglutamate.

[0274] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, and 9, and wherein the N-terminal glutamine or glutamic acid of the antibody variable region has been cyclized to pyroglutamic acid.

[0275] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1, wherein the N-terminal glutamic acid of the antibody variable region has been cyclized to pyroglutamate.

[0276] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1, wherein the N-terminal glutamic acid of the antibody heavy chain variable region is cyclized to pyroglutamic acid.

[0277] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3, wherein the N-terminal glutamic acid of the antibody heavy chain variable region has been cyclized to pyroglutamate.

[0278] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3, wherein the N-terminal glutamic acid of the antibody heavy chain variable region is cyclized to pyroglutamic acid.

[0279] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5, wherein the N-terminal glutamic acid of the antibody heavy chain variable region has been cyclized to pyroglutamate.

[0280] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5, wherein the N-terminal glutamic acid of the antibody heavy chain variable region is cyclized to pyroglutamic acid.

[0281] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7, wherein the N-terminal glutamine of the antibody heavy chain variable region has been cyclized to pyroglutamate.

[0282] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7, and the N-terminal glutamic acid of the antibody variable region is cyclized to pyroglutamic acid.

[0283] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9, wherein the N-terminal glutamic acid of the antibody heavy chain variable region has been cyclized to pyroglutamate.

[0284] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment comprising a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9, wherein the N-terminal glutamic acid of the antibody heavy chain variable region is cyclized to pyroglutamic acid.

[0285] In a further embodiment, the HA in each antibody-drug conjugate can be obtained by expressing in a host cell nucleic acid molecules encoding the heavy and light chains of an antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises: (a) a VH having the amino acid sequence set forth in SEQ ID NO: 1 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 2 or a variant thereof; (b) a VH having the amino acid sequence set forth in SEQ ID NO: 3 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 4 or a variant thereof; (c) a VH having the amino acid sequence set forth in SEQ ID NO: 5 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 6 or a variant thereof; (d) a VH having the amino acid sequence set forth in SEQ ID NO: 7 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 8 or a variant thereof; or (e) a VH having the amino acid sequence set forth in SEQ ID NO: 9 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 10 or a variant thereof; The variant has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity compared to the sequence from which the variant is derived, or has one or several amino acid substitutions, deletions, or additions (e.g., 1, 2, 3, 4, or 5 amino acid substitutions, deletions, or additions) compared to the sequence from which the variant is derived.

[0286] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment thereof obtainable by expressing in a host cell nucleic acid molecules encoding the heavy and light chains of the antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof is (a) a VH having the amino acid sequence set forth in SEQ ID NO: 1, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 2; (b) a VH having the amino acid sequence set forth in SEQ ID NO: 3, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 4; (c) a VH having the amino acid sequence set forth in SEQ ID NO: 5, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 6; (d) a VH having the amino acid sequence set forth in SEQ ID NO: 7, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 8, or (e) VH having the amino acid sequence set forth in SEQ ID NO: 9, and / or VL having the amino acid sequence set forth in SEQ ID NO: 10.

[0287] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment thereof that can be obtained by expressing in a host cell nucleic acid molecules encoding the heavy and light chains of the antibody or antigen-binding fragment thereof, and the antibody or antigen-binding fragment thereof comprises (a) a VH having the amino acid sequence set forth in SEQ ID NO: 1 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 2 or a variant thereof.

[0288] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment thereof obtainable by expressing in a host cell a nucleic acid molecule encoding the heavy and light chains of the antibody or antigen-binding fragment thereof, and the antibody or antigen-binding fragment thereof comprises (b) a VH having the amino acid sequence set forth in SEQ ID NO: 3 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 4 or a variant thereof.

[0289] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment thereof obtainable by expressing in a host cell a nucleic acid molecule encoding the heavy and light chains of the antibody or antigen-binding fragment thereof, and the antibody or antigen-binding fragment thereof comprises (c) a VH having the amino acid sequence set forth in SEQ ID NO: 5 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 6 or a variant thereof.

[0290] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment thereof obtainable by expressing in a host cell a nucleic acid molecule encoding the heavy and light chains of the antibody or antigen-binding fragment thereof, and the antibody or antigen-binding fragment thereof comprises (d) a VH having the amino acid sequence set forth in SEQ ID NO: 7 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 8 or a variant thereof.

[0291] In a further embodiment, the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment thereof obtainable by expressing in a host cell a nucleic acid molecule encoding the heavy and light chains of the antibody or antigen-binding fragment thereof, and the antibody or antigen-binding fragment thereof comprises (e) a VH having the amino acid sequence set forth in SEQ ID NO: 9 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 10 or a variant thereof.

[0292] In some embodiments, the antibody-drug conjugate comprises: [ka] [ka] During the ceremony, The HA in each antibody-drug conjugate was (1) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a VL having the amino acid sequence set forth in SEQ ID NO: 2, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (2) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (3) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a VL having the amino acid sequence set forth in SEQ ID NO: 6, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (4) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a VL having the amino acid sequence set forth in SEQ ID NO: 8, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 8 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; or (5) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a VL having the amino acid sequence set forth in SEQ ID NO: 10, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 10 and a CL having the amino acid sequence set forth in SEQ ID NO: 44, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0293] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in each antibody-drug conjugate was (1) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a VL having the amino acid sequence set forth in SEQ ID NO: 2, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (2) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (3) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a VL having the amino acid sequence set forth in SEQ ID NO: 6, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (4) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a VL having the amino acid sequence set forth in SEQ ID NO: 8, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 8 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (5) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a VL having the amino acid sequence set forth in SEQ ID NO: 10, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 10 and a CL having the amino acid sequence set forth in SEQ ID NO: 44, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0294] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (1) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a VL having the amino acid sequence set forth in SEQ ID NO: 2, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0295] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (2) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a CL having the amino acid sequence set forth in SEQ ID NO: 44, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0296] In a further embodiment, the antibody-drug conjugate is: [ka] the HA in the antibody-drug conjugate is (3) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a VL having the amino acid sequence set forth in SEQ ID NO: 6, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0297] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (4) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a VL having the amino acid sequence set forth in SEQ ID NO: 8, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 8 and a CL having the amino acid sequence set forth in SEQ ID NO: 44, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0298] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (5) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a VL having the amino acid sequence set forth in SEQ ID NO: 10, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 10 and a CL having the amino acid sequence set forth in SEQ ID NO: 44, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0299] In a further embodiment, the antibody-drug conjugate is: [ka] the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44; x is 3 to 4 or 7 to 9; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0300] In a further embodiment, the antibody-drug conjugate is: [ka] HA in each antibody-drug conjugate is an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44, wherein x is 7 to 9; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0301] In a further embodiment, the antibody-drug conjugate is: [ka] the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44, wherein x is 8 to 9; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0302] In a further embodiment, the antibody-drug conjugate is: [ka] the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44, wherein x is 8; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0303] In a further embodiment, the antibody-drug conjugate is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47, wherein x is 3 to 4 or 7 to 9; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0304] In a further embodiment, the antibody-drug conjugate is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47, wherein x is 8 to 9; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0305] In a further embodiment, the antibody-drug conjugate is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47, wherein x is 7 to 8; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0306] In a further embodiment, the antibody-drug conjugate is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47; x is 7 to 8; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via a sulfhydryl of a cysteine ​​in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0307] In a further embodiment, the antibody-drug conjugate is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47, wherein x is 8; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via a sulfhydryl of a cysteine ​​in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0308] In a further embodiment, the antibody-drug conjugate is: [ka] HA is an antibody comprising a heavy chain (HC) consisting of the amino acid sequence set forth in SEQ ID NO: 52, 53, or 54 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, x is 7 to 8; HA is linked via a sulfhydryl of a cysteine ​​in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0309] In a further embodiment, the antibody-drug conjugate is: [ka] HA is an antibody consisting of two heavy chains and two light chains, each heavy chain (HC) consisting of the amino acid sequence set forth in SEQ ID NO: 52, and each light chain (LC) consisting of the amino acid sequence set forth in SEQ ID NO: 47; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, x is 7 to 8; HA is linked via a sulfhydryl of a cysteine ​​in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0310] In a further embodiment, the antibody-drug conjugate is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, wherein: HA is linked via a sulfhydryl of a cysteine ​​in the antibody or antigen-binding fragment to form an antibody-drug conjugate; x is 8.

[0311] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in each antibody-drug conjugate was (1) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a VL having the amino acid sequence set forth in SEQ ID NO: 2, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (2) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (3) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a VL having the amino acid sequence set forth in SEQ ID NO: 6, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (4) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a VL having the amino acid sequence set forth in SEQ ID NO: 8, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 8 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (5) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a VL having the amino acid sequence set forth in SEQ ID NO: 10, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 10 and a CL having the amino acid sequence set forth in SEQ ID NO: 44, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0312] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (1) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a VL having the amino acid sequence set forth in SEQ ID NO: 2, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0313] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (2) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a CL having the amino acid sequence set forth in SEQ ID NO: 44, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0314] In a further embodiment, the antibody-drug conjugate is: [ka] the HA in the antibody-drug conjugate is (3) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a VL having the amino acid sequence set forth in SEQ ID NO: 6, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0315] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in each antibody-drug conjugate is (4) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a VL having the amino acid sequence set forth in SEQ ID NO: 8, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 8 and a CL having the amino acid sequence set forth in SEQ ID NO: 44, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0316] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (5) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a VL having the amino acid sequence set forth in SEQ ID NO: 10, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 10 and a CL having the amino acid sequence set forth in SEQ ID NO: 44, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0317] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in each antibody-drug conjugate is an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3, a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44, x is 7 to 9; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0318] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44. x is 7 to 8; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0319] In a further embodiment, the antibody-drug conjugate is: [ka] the HA in each antibody-drug conjugate is an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44, X is 8, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0320] In a further embodiment, the antibody-drug conjugate is: [ka] the HA in the antibody-drug conjugate is an antibody or an antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44; x is 7 to 9; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0321] In a further embodiment, the antibody-drug conjugate is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47; x is 7 to 8; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0322] In a further embodiment, the antibody-drug conjugate is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47; x is 8, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0323] In a further embodiment, the antibody-drug conjugate is: [ka] HA in antibody-drug conjugates (1) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a VL having the amino acid sequence set forth in SEQ ID NO: 2, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (2) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (3) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a VL having the amino acid sequence set forth in SEQ ID NO: 6, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (4) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a VL having the amino acid sequence set forth in SEQ ID NO: 8, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 8 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (5) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a VL having the amino acid sequence set forth in SEQ ID NO: 10, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 10 and a CL having the amino acid sequence set forth in SEQ ID NO: 44.

[0324] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (1) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a VL having the amino acid sequence set forth in SEQ ID NO: 2, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0325] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (2) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a CL having the amino acid sequence set forth in SEQ ID NO: 44, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0326] In a further embodiment, the antibody-drug conjugate is: [ka] the HA in the antibody-drug conjugate is (3) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a VL having the amino acid sequence set forth in SEQ ID NO: 6, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0327] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in each antibody-drug conjugate is (4) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a VL having the amino acid sequence set forth in SEQ ID NO: 8, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 8 and a CL having the amino acid sequence set forth in SEQ ID NO: 44, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0328] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (5) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a VL having the amino acid sequence set forth in SEQ ID NO: 10, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 10 and a CL having the amino acid sequence set forth in SEQ ID NO: 44, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0329] In a further embodiment, the antibody-drug conjugate is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47, wherein x is 7 to 8; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0330] In a further embodiment, the antibody-drug conjugate is: [ka] HA in antibody-drug conjugates (1) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a VL having the amino acid sequence set forth in SEQ ID NO: 2, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (2) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (3) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a VL having the amino acid sequence set forth in SEQ ID NO: 6, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (4) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a VL having the amino acid sequence set forth in SEQ ID NO: 8, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 8 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (5) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a VL having the amino acid sequence set forth in SEQ ID NO: 10, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 10 and a CL having the amino acid sequence set forth in SEQ ID NO: 44, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0331] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (1) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a VL having the amino acid sequence set forth in SEQ ID NO: 2, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0332] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (2) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a CL having the amino acid sequence set forth in SEQ ID NO: 44, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0333] In a further embodiment, the antibody-drug conjugate is: [ka] the HA in the antibody-drug conjugate is (3) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a VL having the amino acid sequence set forth in SEQ ID NO: 6, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0334] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in each antibody-drug conjugate is (4) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a VL having the amino acid sequence set forth in SEQ ID NO: 8, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 8 and a CL having the amino acid sequence set forth in SEQ ID NO: 44, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0335] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (5) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a VL having the amino acid sequence set forth in SEQ ID NO: 10, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 10 and a CL having the amino acid sequence set forth in SEQ ID NO: 44, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0336] In a further embodiment, the antibody-drug conjugate is: [ka] the HA in the antibody-drug conjugate is an antibody or an antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44; x is 7 to 9; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0337] In a further embodiment, the antibody-drug conjugate is: [ka] the HA in the antibody-drug conjugate is an antibody or an antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 47; x is 7 to 9; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0338] In a further embodiment, the antibody-drug conjugate is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47; x is 7 to 8; [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0339] In a further embodiment, the antibody-drug conjugate is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47; x is 8 to 9, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0340] In a further embodiment, the antibody-drug conjugate is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47; x is 8, [ka] represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and a linker, HA is linked via sulfhydryls in the antibody or antigen-binding fragment to form an antibody-drug conjugate.

[0341] In a further embodiment, the antibody-drug conjugate is: [ka] HA in antibody-drug conjugates (1) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a VL having the amino acid sequence set forth in SEQ ID NO: 2, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (2) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (3) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a VL having the amino acid sequence set forth in SEQ ID NO: 6, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (4) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a VL having the amino acid sequence set forth in SEQ ID NO: 8, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 8 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (5) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a VL having the amino acid sequence set forth in SEQ ID NO: 10, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 10 and a CL having the amino acid sequence set forth in SEQ ID NO: 44.

[0342] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (1) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a VL having the amino acid sequence set forth in SEQ ID NO: 2, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a CL having the amino acid sequence set forth in SEQ ID NO: 44.

[0343] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (2) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a CL having the amino acid sequence set forth in SEQ ID NO: 44.

[0344] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (3) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a VL having the amino acid sequence set forth in SEQ ID NO: 6, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a CL having the amino acid sequence set forth in SEQ ID NO: 44.

[0345] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (4) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a VL having the amino acid sequence set forth in SEQ ID NO: 8, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 8 and a CL having the amino acid sequence set forth in SEQ ID NO: 44.

[0346] In a further embodiment, the antibody-drug conjugate is: [ka] The HA in the antibody-drug conjugate is (5) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a VL having the amino acid sequence set forth in SEQ ID NO: 10, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 10 and a CL having the amino acid sequence set forth in SEQ ID NO: 44.

[0347] In a further embodiment, the antibody-drug conjugate is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47, and x is 2 to 5 or 3 to 4.

[0348] In some embodiments, the antibody-drug conjugate (ADC) has an x ​​of 1 to 10, e.g., 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, 1 to 10, 2 to 3, 2 to 4, 2 to 5, 2 to 6, 2 to 7, 2 to 8, 2 to 9, 2 to 10, 3 to 4, 3 to 5, 3 to 6, 3 to 7, 3 to 8, 3 to 9, 3 to 10, 4 to 5, 4 to 6, 4 to 7, 4 to 8, 4 to 9, 4 to 10, 5 to 6, 5 to 7, 5 to 8, 5 to 9, 5 to 10, 6 to 7, 6 to 8, 6 to 9, 6 to 10, 7 to 8, 7 to 9, 7 to 10, 8 to 9, 8 to 10, or 9 to 10.

[0349] In some embodiments, the antibody-drug conjugate (ADC) has an x ​​of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0350] In some embodiments, the antibodies disclosed herein are engineered to contain one or more cysteine ​​or lysine residues or non-canonical amino acid substitutions of amino acids at defined positions within the antibody.

[0351] In some embodiments, the antibodies disclosed herein are genetically engineered to contain one or more cysteine ​​or non-standard amino acid substitutions of amino acids at defined positions within the antibody. These cysteine ​​or non-standard amino acid residues can then be attached to a drug-linker via the sulfhydryl group of the cysteine ​​residue or the reactive group of the non-standard amino acid. Thus, the antibody-drug conjugates of the present invention can contain one or more substitutions of amino acids in the heavy or light chain of the antibody with cysteine ​​or non-standard amino acid residues, which are then attached to a drug-linker disclosed herein. In certain embodiments, the amino acid positions that can be substituted are selected from positions 152, 153, 171, 172, 173, and 375 (numbered according to the EU numbering scheme) of the heavy chain constant domain and positions 165 and 168 (numbered starting from the N-terminal amino acid 1) of the light chain constant domain. In certain embodiments, cysteine ​​may be substituted for one or more of the amino acids at positions 152, 153, 171, 172, 173, and 375 (numbered according to the EU numbering scheme) of the heavy chain constant domain and positions 165 and 168 (numbered starting from the N-terminus amino acid 1) of the light chain constant domain. In certain embodiments, the antibody-drug conjugate comprises an S375C amino acid substitution attached to a drug-linker disclosed herein. In certain embodiments, the antibody comprises an S375C amino acid substitution and an E152C amino acid substitution, each attached to a drug-linker disclosed herein. In certain embodiments, the antibody comprises an S375C amino acid substitution and an S168C amino acid substitution, each attached to a drug-linker disclosed herein.In some embodiments, as described herein, the ADC compositions may comprise 1-10, e.g., 1-2, 1-3, 1-4, 1-5, 1-6, 1-7, 1-8, 1-9, 1-10, 2-3, 2-4, 2-5, 2-6, 2-7, 2-8, 2-9, 2-10, 3-4, 3-5, 3-6, 3-7, 3-8, 3-9, 3-10, 4-5, 4-6, 4-7, 4-8, 4-9, 4-1 0, 5 to 6, 5 to 7, 5 to 8, 5 to 9, 5 to 10, 6 to 7, 6 to 8, 6 to 9, 6 to 10, 7 to 8, 7 to 9, 7 to 10, 8 to 9, 8 to 10, or 9 to 10, preferably 3 to 9, for example, 3.0 to 3.5, 3.0 to 4.0, 3.0 to 4.5, 3.0 to 5.0, 3.0 to 5.5, 3.0 to 6.0, 3.5 to 4.0, 3.5 to 4.5, 3.5 to 5.0, 3.5 to 5.5, 3.5 to 6.0 , 3.5~6.5, 3.5~7.0, 3.5~7.5, 3.5~8.0, 4.0~4.5, 4.0~5.0, 4.0~5.5, 4.0~6.0, 4.0~6.5, 4.0~7.0, 4.0~7.5, 4.0~8.0, 4.5~5.0, 4.5~5.5, 4.5~6.0, 4.5~6.5, 4.5~7.0, 4.5~7.5, 4.5~8.0, 5.0~5.5, 5.0~6.0, The DAR value (drug-antibody ratio) is 5.0 to 6.5, 5.0 to 7.0, 5.0 to 7.5, 5.0 to 8.0, 5.5 to 6.0, 5.5 to 6.5, 5.5 to 7.0, 5.5 to 7.5, 5.5 to 8.0, 6.0 to 6.5, 6.0 to 7.0, 6.0 to 7.5, 6.0 to 8.5, 6.5 to 7.0, 6.5 to 7.5, 6.5 to 8.5, 7.0 to 7.5, 7.0 to 9.0, or 7.5 to 9.0.

[0352] Drug-Linker Those skilled in the art should understand that the antibody-drug conjugates described herein can be prepared modularly. For example, a free form of a "drug-linker" (which can be understood as M'-L-E, where M' is the structural form of M prior to covalent attachment to an antibody or antigen-binding fragment thereof) can be obtained, and then the free form of the "drug-linker" can be covalently attached to an antibody or antigen-binding fragment thereof to obtain an antibody-drug conjugate according to the present application. Correspondingly, M' in the free form of the "drug-linker" can be linked to one or more sulfhydryls (-SH), aminos (-NH), or carboxyls (-COOH) on an antibody or antigen-binding fragment thereof via substitution reactions (e.g., removal of -SOMe or -Br and similar structures thereon) or addition reactions and other methods.

[0353] In another embodiment, the present invention provides a Drug-Linker having a structure shown as formula M'-LED, M' is [ka] and Lg is a leaving group for nucleophilic substitution reactions (e.g., halogen, methanesulfonyl, fluorophenol, or [ka] ), or hydroxyl (-OH), sulfhydryl group (-SH), or amino (-NH2), or Lg forms an unsaturated double bond with an adjacent atom on ring A, ring A is a 5- to 6-membered aliphatic heterocycle or a 5- to 20-membered aromatic ring system, and the aliphatic heterocycle and aromatic ring system are oxo (=O), halogen, cyano, amino, carboxyl, sulfhydryl, and C 1~6 alkyl, and M is a single bond, C 1~20 Alkylene, C 2~20 Alkenylene, and C 2~20 alkynylene, The L, E, and D structures are defined according to any one of the above antibody-drug conjugates.

[0354] In some embodiments, M' is [ka] Lg is methylsulfonyl, or Lg forms a carbon-carbon double bond together with adjacent atoms on ring A, ring A is a 5-membered alicyclic heterocycle, a 6-membered heteroaromatic ring, or a polycyclic ring formed by connecting two or more 6-membered heteroaromatic rings and a benzene ring with a single bond, and the alicyclic heterocycle is substituted with oxo (=O), halogen, and C 1~4 alkyl, and M is a single bond, C 3~10 Alkylene, C 3~10 Alkenylene, and C 3~10 alkynylene.

[0355] In some embodiments, M' is [ka] and [ka] teeth, [ka] M1 is selected from a single bond, C 5~8 Alkylene, C 5~8 Alkenylene, and C 5~8 alkynylene.

[0356] In some embodiments, M' is [ka] is selected from.

[0357] In some embodiments, M' is [ka] is.

[0358] In some embodiments, the free form of the "drug-linker" is A-01 to A-26, B-01 to B-06, and C-01 shown below: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] is selected from.

[0359] The present invention further provides ADCs comprising an antibody that binds to PTK7 linked via a cysteine ​​or lysine or a non-canonical amino acid substitution of an amino acid within an antibody residue to a drug-linker selected from the group consisting of A-01, A-02, A-03, A-04, A-05, A-06, A-07, A-08, A-09, A-10, A-12, A13, A-14, A15, A-16, A-17, A-18, A-19, A-20, A-21, A-22, A-23, A-24, A-25, A-26, B-01, B-02, B-03, B-04, B-05, B-06, and C-01.

[0360] The present invention further provides ADCs comprising an antibody that binds to PTK7 linked via a cysteine ​​or a non-canonical amino acid substitution of an amino acid within an antibody residue to a drug-linker selected from the group consisting of A-01, A-02, A-03, A-04, A-05, A-06, A-07, A-08, A-09, A-10, A-12, A13, A-14, A15, A-16, A-17, A-18, A-19, A-20, A-21, A-22, A-23, A-24, A-25, A-26, B-01, B-02, B-03, B-04, B-05, B-06, and C-01.

[0361] In a further embodiment of the ADC, the antibody (1) A heavy chain variable region (VH) and / or light chain variable region (VL) of the following, wherein the CDRs are defined according to the Chothia numbering system: (1a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 11 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 12 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof; or (1b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 27 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 28 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof, The variants of either (1a) and (1b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with respect to the sequence from which the variant is derived, or have one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, preferably conservative substitutions, in a heavy chain variable region (VH) and / or a light chain variable region (VL); or (2) A heavy chain variable region (VH) and / or a light chain variable region (VL) of the following, wherein the CDRs are defined according to the Kabat numbering system: (2a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 17 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 18 or 19 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof; or (2b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 33 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 34 or 35 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof; The variants of either (2a) and (2b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with respect to the sequence from which the variant is derived, or have one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, preferably conservative substitutions, in a heavy chain variable region (VH) and / or a light chain variable region (VL); or (3) A heavy chain variable region (VH) and / or light chain variable region (VL) of the following, wherein the CDRs are defined according to the IMGT numbering system: (3a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 20 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 21 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 22 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 23 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 24 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof; or (3b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 36 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 37 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 38 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 39 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 40 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof; The variants of either (3a) and (3b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with respect to the sequence from which the variant is derived, or have one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, and the substitutions are preferably conservative substitutions. and (4) A heavy chain variable region (VH) and / or light chain variable region (VL) of the following, wherein the CDRs are defined according to the AbM numbering system: (4a) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 25 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 26 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof, or (4b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 41 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 42 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof; The variant of either (4a) or (4b) is an antibody or antigen-binding fragment selected from the group consisting of a heavy chain variable region (VH) and / or a light chain variable region (VL) that has at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence from which the variant is derived, or that has undergone one or several amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) with respect to the sequence from which the variant is derived, and preferably the substitutions are conservative substitutions.

[0362] In a further embodiment of the ADC, the antibody or antigen-binding fragment (a) a VH having the amino acid sequence set forth in SEQ ID NO: 1 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 2 or a variant thereof; (b) a VH having the amino acid sequence set forth in SEQ ID NO: 3 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 4 or a variant thereof; (c) a VH having the amino acid sequence set forth in SEQ ID NO: 5 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 6 or a variant thereof; (d) a VH having the amino acid sequence set forth in SEQ ID NO: 7 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 8 or a variant thereof; and

[0363] (e) a VH having the amino acid sequence set forth in SEQ ID NO: 9 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 10 or a variant thereof. In a further embodiment of the ADC, the antibody or antigen-binding fragment is selected from the group consisting of: (1) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a VL having the amino acid sequence set forth in SEQ ID NO: 2, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (2) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (3) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a VL having the amino acid sequence set forth in SEQ ID NO: 6, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (4) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a VL having the amino acid sequence set forth in SEQ ID NO: 8, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 8 and a CL having the amino acid sequence set forth in SEQ ID NO: 44, and (5) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a VL having the amino acid sequence set forth in SEQ ID NO: 10, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 10 and a CL having the amino acid sequence set forth in SEQ ID NO: 44.

[0364] In further embodiments of the ADC, the antibody or antigen-binding fragment includes: (1) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a VL having the amino acid sequence set forth in SEQ ID NO: 2, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a CL having the amino acid sequence set forth in SEQ ID NO: 44.

[0365] In a further embodiment of the ADC, the antibody or antigen-binding fragment includes (2) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a CL having the amino acid sequence set forth in SEQ ID NO: 44.

[0366] In a further embodiment of the ADC, the antibody or antigen-binding fragment includes (3) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a VL having the amino acid sequence set forth in SEQ ID NO: 6, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a CL having the amino acid sequence set forth in SEQ ID NO: 44.

[0367] In a further embodiment of the ADC, the antibody or antigen-binding fragment includes (4) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a VL having the amino acid sequence set forth in SEQ ID NO: 8, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 8 and a CL having the amino acid sequence set forth in SEQ ID NO: 44.

[0368] In a further embodiment of the ADC, the antibody or antigen-binding fragment includes (5) an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a VL having the amino acid sequence set forth in SEQ ID NO: 10, for example, an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 10 and a CL having the amino acid sequence set forth in SEQ ID NO: 44. In a further embodiment, the antibody comprises a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47.

[0369] Antibody preparation The antibodies described herein can be prepared by various methods known in the art, such as genetic engineering and recombinant technology. For example, DNA molecules encoding the heavy and light chain genes of the antibodies of the present invention can be obtained by chemical synthesis or PCR amplification. The resulting DNA molecules are inserted into an expression vector and then transfected into host cells. The transfected host cells are then cultured under specific conditions to express the antibodies of the present invention.

[0370] join In another aspect, the application provides methods for attaching a drug-linker described herein to an antibody described herein to generate an antibody-drug conjugate (ADC) described herein.

[0371] In certain embodiments, an antibody described herein is attached to a drug-linker described herein via a bond to a lysine in the antibody.

[0372] In certain embodiments, the antibodies described herein are linked to the drug-linkers described herein via a bond to a cysteine ​​in the antibody. In certain embodiments, the cysteine ​​is derived from a reduced intrachain disulfide bond in the antibody. In certain embodiments, the cysteine ​​is derived from a reduced interchain disulfide bond in the antibody.

[0373] In some embodiments, the antibody is conjugated to the drug-linker via binding to a reduced interchain disulfide bond in the antibody. For example, an IgG1 antibody comprises four polypeptide chains: two heavy chains comprising VH, CH1, and Fc (e.g., hinge, CH2, and CH3) domains; and two light chains comprising VL and CL domains connected by interchain cysteine ​​disulfide (-SS-) bonds (e.g., two heavy-light chain interchain disulfide bonds and two hinge heavy-heavy chain interchain disulfide bonds). In certain embodiments, when these disulfide bonds are cleaved under reducing conditions, eight reactive cysteine ​​sulfhydryl moieties are generated. In certain embodiments, each of the eight reactive cysteine ​​sulfhydryl moieties is a linker-drug attachment point, allowing up to eight (x=8) drug-linkers to be attached to the reduced antibody. In certain embodiments, any one of the four disulfide bonds is cleaved under reducing conditions, generating two reactive cysteine ​​sulfhydryl moieties. In further embodiments, each of the two reactive cysteine ​​sulfhydryl moieties is a linker-drug attachment point such that two (x=2) linker-drugs can be attached to the reduced antibody. In certain embodiments, any two of the four disulfide bonds are cleaved under reducing conditions, generating four reactive cysteine ​​sulfhydryl moieties. In further embodiments, each of the four reactive cysteine ​​sulfhydryl moieties is a linker-drug attachment point such that four (x=4) linker-drugs can be attached to the reduced antibody. In certain embodiments, any three of the four disulfide bonds are cleaved under reducing conditions, generating six reactive cysteine ​​sulfhydryl moieties. In further embodiments, each of the six reactive cysteine ​​sulfhydryl moieties is a linker-drug attachment point such that six (x=6) linker-drugs can be attached to the reduced antibody.

[0374] In some embodiments, the interchain disulfide bond is between two cysteine ​​residues that are cleaved under reducing conditions, resulting in two reactive cysteine ​​sulfhydryl moieties. In further embodiments, the interchain disulfide bridge in the antibody is between a heavy chain and a light chain, e.g., between C220 of a heavy chain according to EU numbering and C214 of a kappa light chain, or between C220 of a heavy chain according to EU numbering and C214 of a lambda light chain according to Kabat numbering. Additionally, or alternatively, the interchain disulfide bridge in the antibody is between two heavy chains, e.g., between C226 and / or C229 of a first heavy chain and C226 and / or C229 of a second heavy chain according to EU numbering. In some embodiments, the cysteine ​​residues are in the hinge region of the antibody. In some embodiments, the cysteine ​​residue is at any one or more of positions 220, 226, or 229 in the heavy chain according to EU numbering (also referred to herein as C220, C226, or C229, respectively). In some embodiments, the cysteine ​​residue is at position 214 in the light chain according to EU and / or Kabat numbering (also referred to herein as C214, e.g., position 214 in a kappa light chain according to EU and Kabat numbering or position 214 in a lambda light chain according to Kabat numbering). In one embodiment, the cysteine ​​residue is at each of positions 220, 226, and 229 in the heavy chain according to EU numbering and position 214 in the light chain according to EU or Kabat numbering. In one embodiment, the cysteine ​​residue is at each of positions 220, 226, and 229 in the heavy chain according to EU numbering and position 214 in the kappa light chain according to EU and Kabat numbering. In one embodiment, the cysteine ​​residues are at each of positions 220, 226, and 229 of the heavy chain according to EU numbering, and position 214 of the lambda light chain according to Kabat numbering. In one embodiment, the cysteine ​​residues are at any one or more of the following positions: (i) any one, any two, or any three of positions 220, 226, and 229 in the first heavy chain according to EU numbering; (ii) any one, any two, or any three of positions 220, 226, and 229 in the second heavy chain according to EU numbering; (iii) position 214 of the first light chain according to the Kabat numbering system; and / or (iv) Position 214 of the second light chain according to Kabat numbering.

[0375] As used herein, C220, C226, and C229 refer to amino acid residues in immunoglobulins identified according to EU numbering (cysteine, Cys, C). Thus, as will be understood by one of skill in the art, such numbering represents the amino acid residues of a polypeptide aligned with those identified in immunoglobulins, such as those shown at www.imgt.org / IMGTScientificChart / Numbering / Hu_IGHGnber.html.

[0376] As used herein, the cysteine ​​residue at position 214 of the kappa light chain refers to the amino acid residue in immunoglobulins identified according to Kabat numbering (cysteine, Cys, C). Thus, as will be understood by one of skill in the art, such numbering represents the amino acid residues in a polypeptide aligned with those identified in immunoglobulins, such as those shown at www.imgt.org / IMGTScientificChart / Numbering / Hu_IGKCnber.html.

[0377] As used herein, the cysteine ​​residue at position 214 of the lambda light chain refers to the amino acid residue in immunoglobulins identified according to Kabat numbering (cysteine, Cys, C). Thus, as will be understood by one of skill in the art, such numbering represents the amino acid residues in a polypeptide aligned with those identified in immunoglobulins, such as those shown at www.imgt.org / IMGTScientificChart / Numbering / Hu_IGLCnber.html.

[0378] In certain embodiments, the antibodies described herein contain four interchain disulfide bonds in the hinge region that can be reduced, thereby cleaving the bonds and revealing reactive sulfhydryl moieties that can be conjugated with maleimide moieties on a drug-linker, such as the maleimide moieties on the drug-linkers described herein.

[0379] In certain embodiments, the antibodies described herein comprise a lysine residue, the reactive amine side chain of which can be conjugated to a drug-linker, such as a maleimide moiety on a drug-linker described herein.

[0380] In one embodiment, the disclosure provides a method of making an ADC described herein, comprising: a) providing a solution comprising an antibody; b) contacting the solution of a) with a reducing agent; c) contacting the solution of b) with a solution containing a drug-linker or a salt thereof described herein.

[0381] In one embodiment, the reducing agent is tris(2-carboxyethyl)phosphine (TCEP).

[0382] composition In another aspect, the present application provides compositions of antibody-drug conjugates (ADCs) described herein. Such compositions may comprise multiple ADCs described herein, each ADC comprising a drug-linker described herein, where x is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In other words, each antibody molecule in the composition may be conjugated to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 drug-linkers. Thus, compositions can be characterized by a "drug-to-antibody" ratio (DAR) ranging from about 1 to about 10. Methods for determining the DAR are well known to those of skill in the art and include methods using reverse-phase chromatography or HPLC-MS.

[0383] For example, in any embodiment, the ADC compositions described herein may have a β- or β-glucan content of about 1 to about 10, or any subrange therebetween, e.g., about 1 to 2, about 1 to 3, about 1 to 4, about 1 to 5, about 1 to 6, about 1 to 7, about 1 to 8, about 1 to 9, about 1 to 10, about 2 to 3, about 2 to 4, about 2 to 5, about 2 to 6, about 2 to 7, about 2 to 8, about 2 to 9, about 2 to 10, about 3 to 4, The DAR is about 3 to 5, about 3 to 6, about 3 to 7, about 3 to 8, about 3 to 9, about 3 to 10, about 4 to 5, about 4 to 6, about 4 to 7, about 4 to 8, about 4 to 9, about 4 to 10, about 5 to 6, about 5 to 7, about 5 to 8, about 5 to 9, about 5 to 10, about 6 to 7, about 6 to 8, about 6 to 9, about 6 to 10, about 7 to 8, about 7 to 9, about 7 to 10, about 8 to 9, about 8 to 10, or about 9 to 10.

[0384] In certain embodiments, the ADC compositions described herein have a saturation of about 3 to 9, e.g., about 3.0 to 3.5, about 3.0 to 4.0, about 3.0 to 4.5, about 3.0 to 5.0, about 3.0 to 5.5, about 3.0 to 6.0, about 3.5 to 4.0, about 3.5 to 4.5, about 3.5 to 5.0, about 3.5 to 5.5, about 3.5 to 6.0, about 3.5 to 6.5, about 3.5 to 7.0, about 3.5 to 7.5, about 3.5 to 8.0, about 4.0 to 4.5, about 4.0 to 5.0, about 4.0 to 5.5, about 4.0 to 6.0, about 4.0 to 6.5, about 4.0 to 7.0, about 4.0 to 7.5, about 4.0 to 8.0, about 4.5 ...0, about 4.0 to 5.5, about 4.0 to 6.0, about 4.0 to 6.5, about 4.0 to 7.0, about 4.0 to 7. .0, approximately 4.5-5.5, approximately 4.5-6.0, approximately 4.5-6.5, approximately 4.5-7.0, approximately 4.5-7.5, approximately 4.5-8.0, approximately 5.0-5.5, approximately 5.0-6.0, approximately 5.0-6.5, approximately 5.0-7.0, approximately 5.0-7.5, approximately 5.0-8.0, approximately 5.5-6.0, approximately 5.5-6. 5, about 5.5 to 7.0, about 5.5 to 7.5, about 5.5 to 8.0, about 6.0 to 6.5, about 6.0 to 7.0, about 6.0 to 7.5, about 6.0 to 8.5, about 6.5 to 7.0, about 6.5 to 7.5, about 6.5 to 8.5, about 7.0 to 7.5, about 7.0 to 9.0 or about 7.5 to 9.0.

[0385] Pharmaceutical Composition In another aspect, the present application provides a pharmaceutical composition comprising an antibody-drug conjugate (ADC) according to any one of the preceding embodiments and optionally a drug-linker according to any one of the preceding embodiments, and one or more pharmaceutically acceptable adjuvants.

[0386] In one embodiment, the pharmaceutical compositions provided herein comprise an antibody-drug conjugate (ADC) and one or more pharmaceutically acceptable adjuvants, wherein the ADC is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47, and x is 8 to 9.

[0387] In one embodiment, the pharmaceutical compositions provided herein comprise an antibody-drug conjugate (ADC) and one or more pharmaceutically acceptable adjuvants, wherein the ADC is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47, and x is 8.

[0388] In one embodiment, the pharmaceutical compositions provided herein comprise an antibody-drug conjugate (ADC) and one or more pharmaceutically acceptable adjuvants, wherein the ADC is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47, and x is 7 to 8.

[0389] In one embodiment, the pharmaceutical compositions provided herein comprise an antibody-drug conjugate (ADC) and one or more pharmaceutically acceptable adjuvants, wherein the ADC is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47, and x is 8.

[0390] In one embodiment, the pharmaceutical compositions provided herein comprise an antibody-drug conjugate (ADC) and one or more pharmaceutically acceptable adjuvants, wherein the ADC is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47, and x is 7 to 8.

[0391] In one embodiment, the pharmaceutical compositions provided herein comprise an antibody-drug conjugate (ADC) and one or more pharmaceutically acceptable adjuvants, wherein the ADC is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47, and x is 8.

[0392] In one embodiment, the pharmaceutical compositions provided herein comprise an antibody-drug conjugate (ADC) and one or more pharmaceutically acceptable adjuvants, wherein the ADC is:

[0393] In one embodiment, the pharmaceutical compositions provided herein comprise an antibody-drug conjugate (ADC) and one or more pharmaceutically acceptable excipients, wherein the ADC is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47, and x is 8 to 9.

[0394] In one embodiment, the pharmaceutical compositions provided herein comprise an antibody-drug conjugate (ADC) and one or more pharmaceutically acceptable adjuvants, wherein the ADC is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47, and x is 8.

[0395] In one embodiment, the pharmaceutical compositions provided herein comprise an antibody-drug conjugate (ADC) and one or more pharmaceutically acceptable adjuvants, wherein the ADC is: [ka] HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47, and x is 8 to 9.

[0396] Pharmaceutically acceptable auxiliaries include, for example, pharmaceutically acceptable carriers and / or excipients. Pharmaceutically acceptable auxiliaries further include salts and solvates.

[0397] The ADCs described herein are typically formulated with a pharmaceutically acceptable parenteral vehicle to form a unit injectable form for parenteral applications such as bolus injection, intravenous injection, intratumoral injection, etc. Optionally, antibody-drug conjugates having a desired purity are mixed with a pharmaceutically acceptable diluent, carrier, excipient, or stabilizer in the form of a lyophilizate or solution (Remington's Pharmaceutical Sciences (1980) 16). th (Edition, Osol, A. Ed.) The antibody-drug conjugates described herein, or pharmaceutical compositions comprising the antibody-drug conjugates, can be administered via any route appropriate to the individual being treated.

[0398] The ADCs and pharmaceutical compositions described herein can be formulated into any dosage form known in the medical field, such as tablets, pills, suspensions, emulsions, solutions, gels, capsules, powders, granules, elixirs, troches, suppositories, injections (including injections, sterile powders for injection, and concentrated solutions for injection), inhalants, sprays, etc. The preferred dosage form depends on the intended mode of administration and therapeutic use. The pharmaceutical compositions of the present invention must be sterile and stable under the conditions of manufacture and storage. A preferred dosage form is an injection. Such an injection may be a sterile injectable solution. For example, a sterile injectable solution can be prepared by incorporating the required dose of the antibody of the present invention into an appropriate solvent and, optionally, other desired ingredients (including, but not limited to, pH adjusters, surfactants, adjuvants, ionic strength enhancers, etc., osmotic agents, preservatives, diluents, or any combination thereof), followed by filtration sterilization. Additionally, sterile injectable solutions can be prepared as sterile, lyophilized powders (e.g., by vacuum drying or freeze-drying) for ease of storage and use, which can be dispersed in a suitable carrier (e.g., sterile, pyrogen-free water) prior to use.

[0399] Additionally, the ADCs described herein may be present in pharmaceutical compositions in unit dosage form to facilitate administration by any suitable method known in the art, including, but not limited to, oral, oral, sublingual, ophthalmic, topical, parenteral, rectal, intrathecal, intravesical, inguinal, intravesical, topical (e.g., powder, ointment, or drops), or intranasal routes. However, for many therapeutic applications, the preferred route / mode of administration is parenteral (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). Those of skill in the art will appreciate that the route and / or mode of administration will vary depending on the intended purpose. In some preferred embodiments, the ADCs and pharmaceutical compositions described herein are administered by intravenous infusion or injection.

[0400] In certain embodiments, the pharmaceutical composition may further comprise an additional active pharmaceutical agent. In certain embodiments, the additional pharmaceutically active agent is a drug with anti-tumor activity. In certain embodiments, the additional pharmaceutically active agent is selected from the group consisting of an EGFR inhibitor, a HER2 inhibitor, a HER3 inhibitor, a HER4 inhibitor, an IGFR-1 inhibitor, an mTOR inhibitor, a PI3 kinase inhibitor, a PTK inhibitor, a 7c-met or VEGF inhibitor, a chemotherapeutic agent, or any combination thereof. In certain embodiments, the ADC described herein and the additional pharmaceutically active agent are provided as separate components or as a mixed component. Thus, the antibody or antigen-binding fragment thereof of the present invention and the additional pharmaceutically active agent can be administered simultaneously, separately, or sequentially.

[0401] How to use The antibody-drug conjugates described herein, the drug-linkers described herein, or pharmaceutical compositions thereof can be used to treat various diseases or conditions, such as solid tumors or hematological malignancies, e.g., cancers associated with high expression of PTK7, including lung cancer, breast cancer, epidermal cancer (e.g., squamous cell carcinoma of the skin and oral squamous cell carcinoma), ovarian cancer, or esophageal cancer (e.g., esophageal squamous cell carcinoma).

[0402] Thus, the present application provides the use of an antibody-drug conjugate (ADC), a drug-linker, or a pharmaceutical composition containing the same according to any one of the preceding embodiments in the manufacture of a medicament for the treatment of cancer associated with high expression of PTK7.

[0403] At the same time, the present application also provides a method for treating cancer associated with high expression of PTK7, comprising administering to a subject in need thereof a therapeutically effective amount of the antibody-drug conjugate (ADC), drug-linker, or pharmaceutical composition comprising the same described in any one of the preceding embodiments.

[0404] In certain embodiments, the antibody-drug conjugate (ADC), drug-linker, or pharmaceutical composition (e.g., in a subject) (1) Inhibition of cell (e.g., tumor cell) proliferation, (2) inhibition of tumor growth; (3) Induction and / or enhancement of antibody-dependent cellular cytotoxicity, (4) inhibition of PTK7-mediated signaling; (5) Prevention and / or treatment of PTK7-mediated diseases / disorders, or (6) Any combination of the above (1) to (5) is sufficient.

[0405] In certain embodiments, the PTK7-mediated disease / disorder is a tumor, e.g., a PTK7-expressing tumor. In certain embodiments, the tumor is selected from lung cancer, breast cancer, epidermal cancer (e.g., cutaneous squamous cell carcinoma and oral squamous cell carcinoma), ovarian cancer, or esophageal cancer (e.g., esophageal squamous cell carcinoma), or any combination thereof. [Example]

[0406] The following description of specific embodiments further illustrates, but does not limit, the present invention. Those skilled in the art can make various modifications or improvements in accordance with the teachings of the present invention without departing from the principle and scope of the present invention.

[0407] Information about the sequences included in this invention is provided in the table below. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4]

[0408] The abbreviations used herein have the following meanings: [Table 3]

[0409] The structures of the compounds described in the following examples were determined by nuclear magnetic resonance ( 1 The hydroxyl groups were determined by 1 H NMR or mass spectrometry (MS).

[0410] Nuclear magnetic resonance ( 1 H-NMR was performed using a Bruker 400 MHz nuclear magnetic resonance spectrometer. The deuteration reagent was hexadeuterated dimethyl sulfoxide (DMSO-d6), and the internal standard was tetramethylsilane (TMS).

[0411] The abbreviations for nuclear magnetic resonance (NMR) spectra used in the examples are shown below. s: singlet, d: doublet, t: triplet, q: quartet, m: multiplet, br: broad, J: coupling constant, Hz: Hertz, and DMSO-d6: deuterated dimethyl sulfoxide. δ values ​​are expressed in ppm.

[0412] Mass spectra (MS) determinations were performed using an Agilent (ESI) mass spectrometer, model Agilent 6120B.

[0413] Example 1: N-((S)-10-benzyl-1-((1S,9S)-9-ethyl-5-fluoro-9-hydroxyl-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)amino-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-6-(2,5-dioxo-2,5-dihydro-1-H-pyrrol-1-yl)hexanamide (A-01) [ka]

[0414] Compound A-01-1 (0.40 g, 640.59 μmol, for its synthesis see Chinese Patent Application Publication No. 111936169(A)) and exatecan mesylate (0.37 g, 704.65 μmol) were dissolved in DMF (8 mL). HATU (0.32 g, 832.77 μmol) and DIPEA (0.25 g, 1.92 mmol) were added, and the mixture was allowed to react at 25°C for 4 hours. The DIPEA was removed under reduced pressure, and the crude product was lyophilized with water to remove most of the DMF, yielding the title compound (273 mg). This crude product was purified by preparative high-performance liquid chromatography (conditions as follows).

[0415] Chromatography column: Waters XBridge Prep C18 OBD 45mm x 450mm x 8.0μm Mobile phase A: acetonitrile, Mobile phase B: water (0.05% trifluoroacetic acid) [Table 4]

[0416] The structural property data are as follows: ESI-MS m / z: 1034.4[M+H] + .

[0417] Example 2 Synthesis of N-((1S,9S)-5-chloro-9-ethyl-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-hydroxyacetamide and N-((1R,9S)-5-chloro-9-ethyl-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-hydroxyacetamide (1-8-A and 1-8-B) [ka]

[0418] Step 1: Synthesis of 1-chloro-3-bromo-2-methyl-5-nitrobenzene (1-8-2) Compound 1-8-1 (5.00 g, 29.14 mmol) was dissolved in n-heptane (25 mL) at a temperature below 25 °C; concentrated sulfuric acid (25 mL) was added and the mixture was heated to 50 °C. NBS (6.22 g, 34.97 mmol) was added in several portions at 50 °C, and the reaction was continued for 2 hours while maintaining the temperature at 50 °C. The reaction was monitored by thin-layer chromatography (ethyl acetate:petroleum ether = 1:10). After cooling to room temperature, the reaction solution was added dropwise to ice water and extracted with toluene. The organic phases were combined, washed with sodium sulfite solution, water, and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by preparative high-performance liquid chromatography (conditions are shown below). The resulting solution was freeze-dried to obtain 4.88 g of the title compound.

[0419] Chromatography column: C18 ODS 45mm x 450mm x 8.0μm Mobile phase A: acetonitrile, Mobile phase B: water (0.05% formic acid) [Table 5]

[0420] Step 2: Synthesis of 3-chloro-5-bromo-4-methylaniline (1-8-3) Compound 1-8-2 (4.88 g, 19.48 mmol) was dissolved in ethyl acetate (100 mL) at 25 °C, and platinum on carbon (2.00 g, 19.48 mmol, 5% content) was added. After hydrogen substitution, the reaction was carried out under hydrogen protection at 60 °C for 4 hours, and the reaction was monitored by high-performance liquid chromatography-mass spectrometry. The reaction solution was filtered and concentrated to obtain 3.68 g of the crude product of the title compound, which was used directly in the next reaction without further purification.

[0421] Step 3: Synthesis of N-(3-chloro-5-bromo-4-methylphenyl)acetamide (1-8-4) Compound 1-8-3 (3.63 g, 14.82 mmol) was dissolved in ethyl acetate (70 mL) at 20° C.; triethylamine (4.50 g, 44.45 mmol) and acetic anhydride (2.27 g, 22.23 mmol) were added, and the reaction was maintained at 20° C. for 20 hours and monitored by high-performance liquid chromatography-mass spectrometry. Water was added to the reaction mixture. Extraction was performed with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give the crude product, which was slurried in a mixture of ethyl acetate and petroleum ether (1:5) to give 2.86 g of the title compound.

[0422] Step 4: Synthesis of 4-(5-acetamido-3-chloro-2-methylphenyl)but-3-enoic acid (1-8-5) Compound 1-8-4 (1.80 g, 6.86 mmol) was dissolved in THF (20 mL) and water (5 mL) at 20 °C. Vinyl acetic acid (708.31 mg, 8.23 ​​mmol), DIPEA (1.95 g, 15.08 mmol), and tris(o-methylphenyl)phosphorus (62.60 mg, 0.20 mmol) were added, and the reaction system was flushed with nitrogen and then heated to 70 °C for 5 hours. The reaction was monitored by HPLC-MS. 1N sodium hydroxide solution was added to the reaction solution to adjust the pH to 8. Ethyl acetate was added for extraction. The aqueous phase was adjusted to pH 3 with 1N hydrochloric acid and then extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 0.82 g of the title compound, which was used directly in the next reaction.

[0423] Step 5: Synthesis of 4-(5-acetamido-3-chloro-2-methylphenyl)butanoic acid (1-8-6) Compound 1-8-5 (2.60 g, 9.71 mmol) was dissolved in THF (50 mL) at 20 °C, and Pd / C (0.52 g, 10% content) was added. The system was placed under positive hydrogen pressure using a balloon filled with hydrogen gas, and the reaction was carried out at 40 °C for 2 hours. The reaction was monitored by high-performance liquid chromatography-mass spectrometry. The reaction solution was filtered, and the filtrate was concentrated to give 2.43 g of the title compound, which was used directly in the next reaction without further purification.

[0424] Step 6: Synthesis of N-(3-chloro-4-methyl-8-oxo-5,6,7,8-tetrahydronaphth-1-yl)acetamide (1-8-7) Compound 1-8-6 (2.43 g, 9.01 mmol) was dissolved in trifluoroacetic acid (10 mL) and cooled to 5 °C. Trifluoroacetic anhydride (3.78 g, 18.02 mmol, 2.50 mL) was added dropwise. The temperature was maintained at 5 °C and the reaction was allowed to proceed for 4 hours. The reaction was monitored by high-performance liquid chromatography-mass spectrometry. The reaction solution was added to water and dissolved in 10 N sodium hydroxide to adjust the pH to 9. Ethyl acetate was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a silica gel column (ethyl acetate:petroleum ether = 0-20%) to obtain 1.53 g of the title compound.

[0425] Step 7: Synthesis of (Z)-N-(3-chloro-7-(hydroxyimino)-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (1-8-8) Potassium tert-butoxide (1.50 g, 13.37 mmol) was dissolved in THF (16 mL) and tert-butanol (4 mL) at 5 °C. A THF solution (16 mL) of compound 1-8-7 (1.53 g, 6.08 mmol) was added dropwise. After 10 minutes, amyl nitrite (1.14 g, 9.73 mmol) was added dropwise, and the reaction was maintained at 5 °C for 1 hour and monitored by high-performance liquid chromatography-mass spectrometry. The reaction solution was adjusted to pH 5 with 1N hydrochloric acid and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was slurried with methyl tert-butyl ether to give 1.20 g of the title compound.

[0426] Step 8: Synthesis of N-(7-amino-3-chloro-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (1-8-9) Compound 1-8-8 (0.50 g, 1.78 mmol) was dissolved in methyl alcohol (8 mL) and 2N hydrochloric acid (8 mL) at 20 °C. Pd / C (0.15 g, 10% content) was added. The system was placed under positive hydrogen pressure using a balloon filled with hydrogen gas, and the reaction was carried out at 5 °C for 2 hours. The reaction was monitored by high-performance liquid chromatography-mass spectrometry. The reaction solution was filtered and concentrated to give 0.52 g of the hydrochloride salt of the title compound, which was used directly in the next reaction without further purification.

[0427] Step 9: Synthesis of N,N'-(3-chloro-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1,7-diyl)diacetamide (1-8-10) Compound 1-8-9 (0.52 g, 1.70 mmol) was dissolved in pyridine (5 mL) at 20°C. Acetic anhydride was added (2 mL). The reaction was maintained at 20°C for 2 hours and monitored by high-performance liquid chromatography-mass spectrometry. Water was added to the reaction solution. Ethyl acetate was added for extraction. The organic layers were washed with water, combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a silica gel column (ethyl acetate:petroleum ether = 0 to 30%) to give 0.22 g of the title compound.

[0428] Step 10: Synthesis of N-(8-amino-6-chloro-5-methyl-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)acetamide (1-8-11) Compound 1-8-10 (450.97 mg, 1.46 mmol) was dissolved in methanol (16 mL) at 20 ° C., 2N hydrochloric acid (16 mL) was added, and the mixture was heated at 60 ° C. for 2 hours. The reaction was monitored by high-performance liquid chromatography-mass spectrometry. Saturated sodium bicarbonate solution was added to the cooled reaction solution to adjust the pH to 8. Ethyl acetate was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 230.00 mg of the title compound, which was used directly in the next step without further purification.

[0429] Step 11: Synthesis of N-((9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzopyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)propyl)acetamide (1-8-12) Compound 1-8-11 (230.00 mg, 0.78 mmol) was dissolved in toluene (10 mL), and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3610(4H)-trione (230.00 mg, 0.87 mmol) and p-toluenesulfonic acid (26.73 mg, 0.16 mmol) were added. The mixture was heated to 140 °C and reacted for 5 hours. The reaction was monitored by high-performance liquid chromatography-mass spectrometry. The reaction solution was concentrated, and the crude product was purified on a silica gel column (methanol:dichloromethane = 0 to 10%) to obtain 150.00 mg of the title compound.

[0430] Step 12: Synthesis of (9S)-1-amino-5-chloro-9-ethyl-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzopyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (1-2) Compound 1-8-12 (40.00 mg, 0.081 mmol) was added to concentrated hydrochloric acid (1 mL) and heated at 100 °C for 5 hours. The reaction was monitored by high-performance liquid chromatography-mass spectrometry. The reaction solution was filtered, and the filtrate was purified by preparative high-performance liquid chromatography (under the following conditions). The resulting solution was lyophilized to obtain the hydrochloride salt of title compound 1-2. Compound 1-2 hydrochloride was separated under the following purification conditions to give two isomers, designated 1-2-A (5.00 mg trifluoroacetic acid, retention time 9.85 min) and 1-2-B (7.00 mg trifluoroacetic acid, retention time 10.62 min), according to their retention times.

[0431] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Atolytic acid, mobile phase B: water (0.05% Tonitric acid)

Table 6

[0432] Structural characteristics are as follows: 1-2-A: 1 H-NMR(400MHz,DMSO-d6)δ 8.42(s,3H),8.27(s,1H),7.36(s,1H),6.59(s,1H),5.78-5.63(m,1H),5.50-5.36(m,3H),5.10-5.06 (m,1H),3.20-3.04(m,2H),2.56(s,3H),2.26-2.13(m,2H),1.93-1.79(m,2H),0.88(t,J=7.2Hz,3H). ESI-MS (m / z): 452.1[M+H] + . 1-2-B: 1 H-NMR(400MHz,DMSO-d6)δ8.42(s,3H),8.27(s,1H),7.36(s,1H),6.58(s,1H),5.78-5.63(m,1H),5.50-5.36(m,3H ),5.10-5.06(m,1H),3.20-3.04(m,2H),2.55(s,3H),2.26-2.13(m,2H),1.93-1.79(m,2H),0.88(t,J=7.2Hz,3H). ESI-MS m / z: 452.0[M+H] + .

[0433] Step 13: 2-((tert-butyldiphenylsilyl)oxy)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxyl-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4': Synthesis of 6,7]indolazino[1,2-b]quinolin-1-yl)acetamide and 2-((tert-butyldiphenylsilyl)oxy)-N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolazino[1,2-b]quinolin-1-yl)acetamide (1-8-13-A and 1-8-13-B) The hydrochloride salt of compound 1-2 (40.00 mg, 81.91 μmol) was dissolved in N,N-dimethylformamide (1 mL) at 25 °C. 2-((tert-butyldiphenylsilyl)oxy)acetic acid (30.91 mg, 98.29 μmol), HATU (62.25 mg, 163.81 μmol), and N,N-diisopropylethylamine (42.34 mg, 327.63 μmol) were added, and the reaction mixture was maintained at 25 °C for 0.5 h and monitored by high-performance liquid chromatography-mass spectrometry. After the reaction was completed, water was added to the reaction solution. Dichloromethane / methanol (v / v = 10 / 1) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified and separated by preparative thin-layer chromatography (dichloromethane:methanol=20:1). Two isomers were named 1-8-13-A (15.00 mg, Rf value 0.3) and 1-8-13-B (12.00 mg, Rf value 0.35) according to their Rf values.

[0434] Step 14: Synthesis of N-((1S,9S)-5-chloro-9-ethyl-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-hydroxyacetamide and N-((1R,9S)-5-chloro-9-ethyl-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-hydroxyacetamide (1-8-A and 1-8-B) 1-8-13-A (15.00 mg) and 1-8-13-B (12.00 mg) were dissolved in tetrahydrofuran (1 mL) in two reaction flasks at 25 °C. Tetrabutylammonium fluoride (1 M in tetrahydrofuran) / glacial acetic acid mixture (v / v = 13 / 1) (50 μL) was added, and the reaction mixture was maintained at 25 °C for 0.5 hours and monitored by high-performance liquid chromatography-mass spectrometry. After completion of the reaction, the reaction solution was purified by preparative high-performance liquid chromatography, and the resulting solution was lyophilized to give the title compounds 1-8-A (i.e., 1-10) (6.94 mg) and 1-8-B (i.e., 1-9) (4.00 mg), respectively.

[0435] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: acetonitrile, Mobile phase B: water (0.05% formic acid) [Table 7]

[0436] The structural property data of 1-8-A (i.e., 1-10) are as follows: 1H NMR(400MHz,DMSO-d6)δ8.43(d,J=8.8Hz,1H),8.16(s,1H),7.31(s,1H),6.55(s,1H),5.65-5.36(m,4H),5.21(q,J=19.0H z,2H),3.95(d,J=5.7Hz,2H),3.26-3.11(m,2H),2.53(s,3H),2.30-2.08(m,2H),1.94-1.79(m,2H),0.87(t,J=7.3Hz,3H). ESI-MS (m / z): 510.1 [M+H] + .

[0437] The structural characteristics of 1-8-B (すなわち, 1-9) are as follows: 1H NMR(400MHz,DMSO-d6)δ8.45(d,J=8.9Hz,1H),8.15(s,1H),7.31(s,1H),6.54(s,1H),5.64-5.35(m,4H),5.19(q,J=19.0H z,2H),3.97(d,J=5.2Hz,2H),3.27-3.10(m,2H),2.51(s,3H),2.27-2.10(m,2H),1.93-1.80(m,2H),0.88(t,J=7.3Hz,3H). ESI-MS (m / z): 510.1[M+H] + .

[0438] Example 3: N-((10S)-10-benzyl-1-(((1S,9S)-5-chloro-9-ethyl-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-6-(2-(methanesulfonyl)pyrimidin-5-yl)hexa-5-ynamide and N-((10S)-10-benzyl-1-(((1R,9S)-5-chloro-9-ethyl-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)amino)-1,6,9,12,15-tetraoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-6-(2-(methylsulfonyl)pyrimidin-5-yl)hexa-5-ynamide (A-07-A (A-05) and A-07-B (A-06)) [ka]

[0439] Step 1: Synthesis of (S)-10-benzyl-23-(2-(methylsulfonyl)pyrimidin-5-yl)-6,9,12,15,18-pentaoxo-3-oxa-5,8,11,14,17-pentaazatricosane-22-ynecarboxylic acid (A-07-3) Compound A-07-2 (30.00 mg, 0.07 mmol) was dissolved in DMF (0.2 mL) at 25 ° C. 2,5-Dioxopyrrolidin-1-yl-6-(2-(methanesulfonyl)pyrimidin-5-yl)hexyne-5-oate ester (A-07-1, 28.00 mg, 0.08 mmol) was added and the reaction was carried out at 30 ° C. for 1 hour. The reaction was monitored by high-performance liquid chromatography-mass spectrometry. The reaction solution was directly purified by preparative high-performance liquid chromatography (conditions are as follows), and the resulting solution was lyophilized to obtain 20.00 mg of the title compound.

[0440] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: acetonitrile, Mobile phase B: water (0.05% formic acid) [Table 8]

[0441] The structural property data are as follows: ESI-MS (m / z): 691.0 [M+HO] + .

[0442] Step 2: N-((10S)-10-benzyl-1-(((1S,9S)-5-chloro-9-ethyl-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-6-(2-(methanesulfonyl)pyrimidin-5-yl)hexan-5-amide and N-((1 Synthesis of 0S)-10-benzyl-1-(((1R,9S)-5-chloro-9-ethyl-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-6-(2-(methanesulfonyl)pyrimidin-5-yl)hexa-5-amide (A-07-A and A-07-B) The hydrochloride salt of 1-2 (30.00 mg, 61.43 μmol) was dissolved in N,N-dimethylformamide (1 mL) at 25 °C. A-07-3 (49.66 mg, 73.72 μmol), HATU (35.01 mg, 92.14 μmol), and N,N-diisopropylethylamine (23.82 mg, 184.29 μmol) were added sequentially, and the reaction was maintained at 25 °C for 0.5 h and monitored by high-performance liquid chromatography-mass spectrometry. After completion of the reaction, the reaction solution was purified by preparative high-performance liquid chromatography (under the following conditions), and the resulting solution was lyophilized to give the title compound A-07. A-07 was separated under the following purification conditions to give two isomers, which were named A-07-A (i.e., A-05) (11.04 mg, retention time 7.5 min) and A-07-B (i.e., A-06) (19.42 mg, retention time 8.0 min) according to their retention times.

[0443] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: acetonitrile, Mobile phase B: water (0.05% formic acid) [Table 9]

[0444] The structural property data are as follows:

[0445] A-07-A (i.e., A-05): ESI-MS (m / z): 1107.3[M+H] + . 1H NMR(400MHz,DMSO)δ9.10(s,2H),8.66-8.63(m,1H),8.51(d,J=8.8Hz,1H),8.34-8.31(m,1H),8.21-8.19(m,1H),8.17-8.09(m,2 H),8.08-8.04(m,1H),7.30(s,1H),7.26-7.15(m,5H),6.55(s,1H),5.56-5.55(m,1H),5.48-5.35(m,2H),5.25-5.10(m,2H),4.64 (d,J=6.4Hz,2H),4.45-4.44(m,1H),4.06-3.98(m,2H),3.77--3.52(m,6H),3.41(s,3H),3.25-3.12(m,2H),3.03-3.00(m,1H),2 .83-2.72(m,1H),2.58-2.56(m,2H),2.48(s,3H),2.33-2.30(m,2H),2.21-2.13(m,2H),1.91-1.76(m,4H),0.87(t,J=7.2Hz,3H).

[0446] A-07-B (i.e., A-06): ESI-MS (m / z): 1107.3[M+H] + .

[0447] Example 4: (S)-7-Ethyl-7-hydroxy-14-(2-(isopropylamino)ethyl)-10,13-dihydro-11H-[1,3]dioxolo[4,5-g]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-8,11(7H)-dione (2-2) [ka]

[0448] Step 1: 3-(isopropylamino)-1-(6-nitrobenzo[d][1,3]dioxin-5-yl)propan-1-one (2-2-2) Compound 2-2-1 (1.90 g, 8.08 mmol) was slowly added to nitric acid (8 mL) at 0° C., and the temperature was slowly raised to 25° C. for 1 hour. The reaction solution was directly purified by reverse-phase column chromatography (acetonitrile / 0.5% formic acid aqueous solution) to obtain 1.50 g of the formate salt of the title compound.

[0449] The structural property data are as follows: ESI-MS (m / z): 281.1[M+H] + .

[0450] Step 2: Synthesis of 1-(6-aminobenzo[d][1,3]dioxin-5-yl)-3-(isopropylamino)propan-1-one (2-2-3) Compound 2-2-2 formate (1.25 g, 4.46 mmol) was added to tetrahydrofuran (20 mL). 10% palladium carbon (125.00 mg) was added. The atmosphere was purged with hydrogen three times, and then the reaction was carried out at 25°C for 16 hours. The reaction solution was filtered through Celite, and the filtrate was concentrated to dryness under reduced pressure to obtain 895.00 mg of the crude product of the title compound, which was used directly in the next reaction without purification.

[0451] The structural property data are as follows: ESI-MS (m / z): 251.1[M+H] + .

[0452] Step 3: Synthesis of (S)-7-ethyl-7-hydroxy-14-(2-(isopropylamino)ethyl)-10,13-dihydro-11H-[1,3]dioxolo[4,5-g]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-8,11(7H)-dione (2-2) Compound 2-2-3 (23.00 mg, 0.09 mmol) and (4S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (21.00 mg, 0.09 mmol) were dissolved in toluene (4 mL). p-Toluenesulfonic acid (1.40 mg, 0.009 mmol) was added, and the mixture was reacted at 140 °C for 12 hours. The solvent was removed under reduced pressure to obtain the crude product of the title compound. The crude product was purified by HPLC (mobile phase A: acetonitrile, mobile phase B: 0.05% aqueous formic acid). Three drops of 3 M hydrochloric acid were added to the prepared solution, followed by lyophilization to obtain 8.70 mg of the hydrochloride salt of the title compound.

[0453] The structural property data are as follows: ESI-MS(m / z):478.2[M+H] + . 1 H-NMR(400MHz,DMSO-d6): δ 9.38(brs,2H),7.85(s,1H),7.52(s,1H),7.25(s,1H),6.30(d,J=2.0Hz,2H),5.43(s,2H),5.30(s,2H),3.57-3. 45(m,2H),3.40-3.25(m,1H),3.22-3.06(m,2H),1.95-1.77(m,2H),1.27(d,J=6.4Hz,6H),0.87(t,J=7.6Hz,3H).

[0454] Example 5: 4-((S)-2-(4-aminobutyl)-35-(4-((6-(2-(methanesulfonyl)pyrimidin-5-yl)hex-5-ynamido)methyl)-1H-1,2,3-triazol-1-yl)-4,8-dioxo-6,12,15,18,21,24,27,30,33-nonaoxa-3,9-diazapentatriacontanamido)benzyl ((S)-4-ethyl-11-(2-(N-isopropylmethylsulfonamido)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)ethyl)carbonate (B-01) [ka]

[0455] Step 1: Synthesis of (S)-4-ethyl-11-(2-(N-isopropylmethanesulfonamido)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7-indolizino[1,2-b]quinolin-4-yl(4-((S)-2-(4-(((4-methyloxyphenyl)diphenylmethyl)amino)butyl)-35-(4-((6-(2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynamido)methyl)-1H-1,2,3-triazol-1-yl)-4,8-dioxo-6,12,15,18,21,24,27,30,33-nonaoxa-3,9-diazapentatriacontanylamido)benzyl)carbonate (B-01-2) Compound B-01-1 (413.40 mg, 0.251 mmol, its synthesis is described in Patent CN111295389B) was dissolved in dimethyl sulfoxide and water (2.0 mL: 0.5 mL) at room temperature. Cuprous (I) (72.95 mg, 0.503 mmol) and 6-(2-(methylsulfonyl)pyrimidin-5-yl)-N-(prop-2-yn-1-yl)-hex-5-ynamide (95.10 mg, 0.302 mmol) were added, stirred for 1 hour, and then filtered. The filtrate was purified by preparative high-performance liquid chromatography (conditions are as follows) to obtain 30.00 mg of the title compound.

[0456] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: acetonitrile, Mobile phase B: water [Table 10]

[0457] The structural property data are as follows: ESI-MS(m / z):815.9[(M-273) / 2+H] + .

[0458] Step 2: Synthesis of 4-((S)-2-(4-aminobutyl)-35-(4-((6-(2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynamido)methyl)-1H-1,2,3-triazol-1-yl)-4,8-dioxo-6,12,15,18,21,24,27,30,33-nonaoxa-3,9-diazapentatriacontanylamido)benzyl ((S)-4-ethyl-11-(2-(N-isopropylmethylsulfonamido)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)ethyl)carbonate (B-01) Compound B-01-3 (30.00 mg, 0.02 mmol) was dissolved in dichloromethane (1.0 mL), and trifluoroacetic acid (0.2 mL) was added to the reaction solution, followed by reaction at room temperature for 30 minutes. The reaction solution was concentrated under reduced pressure and purified by preparative high performance liquid chromatography (conditions are as follows) to obtain 20.00 mg of the trifluoroacetic acid salt of the title compound.

[0459] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: acetonitrile, Mobile phase B: water (0.05% trifluoroacetic acid) [Table 11]

[0460] The structural characterization data is as follows: ESI-MS (m / z): 508.2 [M+H] + .

[0461] Example 6 Synthesis of (2S,3S,4S,5R,6S)-6-(4-(((2-((S)-7-ethyl-7-hydroxy-8,11-dioxo-7,8,11,13-tetrahydro-10H-[1,3]dioxolano[4,5-g]pyrano[3',4':6,7]-indolizino[1,2-b]quinolin-14-yl)ethyl)(isopropyl)carbamoyl)oxy)methyl)-2-(2-(2-(2-(6-(2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynamido)ethoxy)ethoxy)acetamido)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (B-03) [ka]

[0462] Step 1: Synthesis of (2S,3R,4S,5S,6S)-2-(4-(hydroxymethyl)-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triacetate ester (B-03-3) The compound (2R,3R,4S,5S,6S)-2-bromo-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triacetic acid triester (B-03-1, 12.32 g, 31.02 mmol) and 4-hydroxy-3-nitrobenzyl alcohol (compound B-03-2, 5.00 g, 29.56 mmol) were dissolved in acetonitrile (200 mL). Silver oxide (27.40 g, 118.25 mmol) was added with stirring, and the mixture was purged with nitrogen and reacted at room temperature in the dark for 12 hours. The reaction was monitored by high-performance liquid chromatography-mass spectrometry. The reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure and then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:3) to obtain 12.80 g of the title compound.

[0463] The structural property data are as follows: ESI-MS m / z: 503[M+18] + .

[0464] Step 2: Synthesis of (2S,3R,4S,5S,6S)-2-(2-amino-4-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triacetate ester (B-03-4) Compound B-03-3 (2.20 g, 4.53 mmol) was dissolved in ethyl acetate and tetrahydrofuran (50 mL each). PtO (0.20 g) was added, and the reaction system was then purged with a hydrogen balloon three times and reacted under a hydrogen atmosphere for 2 hours. The reaction was monitored by high-performance liquid chromatography-mass spectrometry. The reaction solution was directly filtered, the filter cake was rinsed with ethyl acetate, and the filtrate was evaporated to dryness under reduced pressure to obtain 2.02 g of the crude product of the title compound, which was used directly in the next reaction.

[0465] The structural property data are as fo...

Claims

1. Formula Ab-[M-L-ED] ｘ An antibody-drug conjugate having the structure shown in Ab is an antibody or antigen-binding fragment thereof that specifically binds to human PTK7; M is a linker linked to the antibody or antigen-binding fragment thereof; L is a linker between M and E; E is a structural fragment connecting L and D; D is a cytotoxic drug fragment; An antibody-drug conjugate wherein x is selected from 1-10.

2. the antibody or antigen-binding fragment thereof (1) The following heavy chain variable region (VH) and / or light chain variable region (VL), wherein the CDRs are defined according to the Chothia numbering system: (1a) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence shown as SEQ ID NO: 11 or a variant thereof, CDR-H2 having the sequence shown as SEQ ID NO: 12 or a variant thereof, and CDR-H3 having the sequence shown as SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence shown as SEQ ID NO: 14 or a variant thereof, CDR-L2 having the sequence shown as SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the sequence shown as SEQ ID NO: 16 or a variant thereof; or (1b) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence shown as SEQ ID NO: 27 or a variant thereof, CDR-H2 having the sequence shown as SEQ ID NO: 28 or a variant thereof, and CDR-H3 having the sequence shown as SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence shown as SEQ ID NO: 30 or a variant thereof, CDR-L2 having the sequence shown as SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the sequence shown as SEQ ID NO: 32 or a variant thereof; The variant of any of (1a) and (1b) has a heavy chain variable region (VH) and / or a light chain variable region (VL) that has at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence from which the variant is derived, or that has one or several amino acid substitutions, deletions, or additions with the sequence from which the variant is derived; or (2) The following heavy chain variable region (VH) and / or light chain variable region (VL): (2a) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence shown as SEQ ID NO: 17 or a variant thereof, CDR-H2 having the sequence shown as SEQ ID NO: 18 or 19 or a variant thereof, and CDR-H3 having the sequence shown as SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence shown as SEQ ID NO: 14 or a variant thereof, CDR-L2 having the sequence shown as SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the sequence shown as SEQ ID NO: 16 or a variant thereof; or (2b) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence shown as SEQ ID NO: 33 or a variant thereof, CDR-H2 having the sequence shown as SEQ ID NO: 34 or 35 or a variant thereof, and CDR-H3 having the sequence shown as SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence shown as SEQ ID NO: 30 or a variant thereof, CDR-L2 having the sequence shown as SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the sequence shown as SEQ ID NO: 32 or a variant thereof; The variant of any of (2a) and (2b) has a heavy chain variable region (VH) and / or a light chain variable region (VL) that has at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence from which the variant is derived, or that has one or several amino acid substitutions, deletions, or additions with the sequence from which the variant is derived; or (3) The following heavy chain variable region (VH) and / or light chain variable region (VL): wherein the CDRs are defined according to the IMGT numbering system: (3a) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence shown as SEQ ID NO: 20 or a variant thereof, CDR-H2 having the sequence shown as SEQ ID NO: 21 or a variant thereof, and CDR-H3 having the sequence shown as SEQ ID NO: 22 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence shown as SEQ ID NO: 23 or a variant thereof, CDR-L2 having the sequence shown as SEQ ID NO: 24 or a variant thereof, and CDR-L3 having the sequence shown as SEQ ID NO: 16 or a variant thereof; or (3b) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence shown as SEQ ID NO: 36 or a variant thereof, CDR-H2 having the sequence shown as SEQ ID NO: 37 or a variant thereof, and CDR-H3 having the sequence shown as SEQ ID NO: 38 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence shown as SEQ ID NO: 39 or a variant thereof, CDR-L2 having the sequence shown as SEQ ID NO: 40 or a variant thereof, and CDR-L3 having the sequence shown as SEQ ID NO: 32 or a variant thereof; The variant of any of (3a) and (3b) has a heavy chain variable region (VH) and / or a light chain variable region (VL) that has at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence from which the variant is derived, or that has one or several amino acid substitutions, deletions, or additions with the sequence from which the variant is derived; or (4) The following heavy chain variable region (VH) and / or light chain variable region (VL), wherein the CDRs are defined according to the AbM numbering system: (4a) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence shown as SEQ ID NO: 25 or a variant thereof, CDR-H2 having the sequence shown as SEQ ID NO: 26 or a variant thereof, and CDR-H3 having the sequence shown as SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence shown as SEQ ID NO: 14 or a variant thereof, CDR-L2 having the sequence shown as SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the sequence shown as SEQ ID NO: 16 or a variant thereof; or (4b) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence shown as SEQ ID NO: 41 or a variant thereof, CDR-H2 having the sequence shown as SEQ ID NO: 42 or a variant thereof, and CDR-H3 having the sequence shown as SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence shown as SEQ ID NO: 30 or a variant thereof, CDR-L2 having the sequence shown as SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the sequence shown as SEQ ID NO: 32 or a variant thereof; The antibody-drug conjugate of claim 1, wherein the variant of either (4a) or (4b) comprises a heavy chain variable region (VH) and / or a light chain variable region (VL) that has at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity compared to the sequence from which the variant is derived, or that has one or several amino acid substitutions, deletions, or additions compared to the sequence from which the variant is derived.

3. the antibody or antigen-binding fragment thereof (a) a VH as set forth in SEQ ID NO: 1 or a variant thereof, and / or a VL as set forth in SEQ ID NO: 2 or a variant thereof; (b) a VH as set forth in SEQ ID NO: 3 or a variant thereof, and / or a VL as set forth in SEQ ID NO: 4 or a variant thereof; (c) comprising a VH set forth as SEQ ID NO: 5 or a variant thereof, and / or a VL set forth as SEQ ID NO: 6 or a variant thereof; (d) a VH set forth as SEQ ID NO: 7 or a variant thereof, and / or a VL set forth as SEQ ID NO: 8 or a variant thereof; or (e) comprising a VH set forth as SEQ ID NO: 9 or a variant thereof, and / or a VL set forth as SEQ ID NO: 10 or a variant thereof; The antibody-drug conjugate of claim 1 or 2, wherein the variant has at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity compared to the sequence from which the variant is derived, or has one or several amino acid substitutions, deletions, or additions compared to the sequence from which the variant is derived.

4. the antibody or antigen-binding fragment thereof (a) a human immunoglobulin heavy chain constant region (CH) or a variant thereof, wherein the variant has one or more amino acid substitutions, deletions, or additions compared to the wild-type sequence from which the variant is derived; and The antibody-drug conjugate of any one of claims 1 to 3, further comprising (b) a human immunoglobulin light chain constant region (CL) or a variant thereof, wherein the variant has one or more amino acid substitutions, deletions, or additions compared to the wild-type sequence from which the variant is derived.

5. the antibody or antigen-binding fragment thereof (1) A heavy chain comprising a VH sequence shown as SEQ ID NO: 1 and a heavy chain constant region (CH) shown as SEQ ID NO: 43, and a light chain comprising a VL sequence shown as SEQ ID NO: 2 and a light chain constant region (CL) described in SEQ ID NO: 44; (2) A heavy chain comprising a VH sequence shown as SEQ ID NO: 3 and a heavy chain constant region (CH) shown as SEQ ID NO: 43 or 45, and a light chain comprising a VL sequence shown as SEQ ID NO: 4 and a light chain constant region (CL) described in SEQ ID NO: 44; (3) A heavy chain comprising a VH sequence shown as SEQ ID NO: 5 and a heavy chain constant region (CH) shown as SEQ ID NO: 43, and a light chain comprising a VL sequence shown as SEQ ID NO: 6 and a light chain constant region (CL) described in SEQ ID NO: 44; (4) A heavy chain comprising the VH sequence shown as SEQ ID NO: 7 and the heavy chain constant region (CH) shown as SEQ ID NO: 43, and a light chain comprising the VL sequence shown as SEQ ID NO: 8 and the light chain constant region (CL) described in SEQ ID NO: 44; or (5) The antibody-drug conjugate according to any one of claims 1 to 4, comprising a heavy chain comprising the VH sequence shown as SEQ ID NO: 9 and the heavy chain constant region (CH) shown as SEQ ID NO: 43, and a light chain comprising the VL sequence shown as SEQ ID NO: 10 and the light chain constant region (CL) described in SEQ ID NO:

44.

6. M, 【Chemical 1】 and Ring A is a 5- to 6-membered aliphatic heterocycle or a 5- to 20-membered aromatic ring system, and the aliphatic heterocycle and the aromatic ring system are each independently selected from the group consisting of oxo (=O), halogen, cyano, amino, carboxyl, mercapto, and C １～６ alkyl; and M １ is a single bond, C １～２０ Alkylene, C ２～２０ Alkenylene, and C ２～２０ The antibody-drug conjugate of any one of claims 1 to 5, wherein the alkynylene is selected from the group consisting of alkynylene.

7. M, 【Chemistry 2】 wherein ring A is a 5-membered aliphatic heterocycle, a 6-membered aromatic heterocycle, or a polycyclic ring formed by connecting two or more 6-membered aromatic heterocycles and a benzene ring via a single bond, and the aliphatic heterocycle is selected from the group consisting of oxo (=O), halogen, and C １～４ alkyl; and M １ is a single bond, C ３～１０ Alkylene, C ３～１０ Alkenylene, and C ３～１０ The antibody-drug conjugate of any one of claims 1 to 5, wherein the aryl group is selected from the group consisting of alkynylene.

8. M, 【Chemistry 3】 wherein the ring A is 【Chemistry 4】 is selected from M １ is a single bond, C ５～８ Alkylene, C ５～８ Alkenylene, and C ５～８ The antibody-drug conjugate of any one of claims 1 to 5, wherein the alkynylene is selected from the group consisting of alkynylene.

9. M, 【Chemistry 5】 The antibody-drug conjugate according to any one of claims 1 to 5, wherein the antibody-drug conjugate is selected from the group consisting of:

10. M, 【Chemistry 6】 The antibody-drug conjugate according to any one of claims 1 to 5, wherein

11. L is C １～６ Alkylene, -N(R')-, carbonyl, -O-, Val, Cit, Phe, Lys, Lys(COCH ２ CH ２ (OCH ２ CH ２ ) ｓ OCH ３ ), D-Val, Leu, Gly, Ala, Asn, Val-Cit, Val-Ala, Val-Lys, Val-Lys(Ac), Phe-Lys, Phe-Lys(Ac), D-Val-Leu-Lys, Gly-Gly-Arg, Ala-Ala-Asn, Ala-Ala-Ala, Val-Lys-Ala, Val-Lys-Gly, Gly-Gly-Gly, Gly-Gly-Phe-Gly (SEQ ID NO: 48), Gly-Phe-Leu-Gly (SEQ ID NO: 49), Gly-Gly-Val-Ala (SEQ ID NO: 50), Gly-Gly-Gly-Gly-Gly (SEQ ID NO: 51), 【Chemistry 7】 and R' is selected from the group consisting of hydrogen, C １～６ Alkyl, or -(CH ２ CH ２ 11. The antibody-drug conjugate of claim 1, wherein R represents an alkyl group containing r-, r is an integer selected from 1 to 10, and s is an integer selected from 1 to 20.

12. L is C １～６ Alkylene, -NH-, Val, Cit, Phe, Lys, Lys(COCH ２ CH ２ (OCH ２ CH ２ ) ｓ OCH ３ ), Gly, Val-Cit, Gly-Gly-Phe-Gly (SEQ ID NO: 48), 【Chemistry 8】 and s is an integer selected from 1 to 20.

13. L has the following structure: 【Chemistry 9】 is selected from the group consisting of The antibody-drug conjugate of any one of claims 1 to 10, wherein s is an integer selected from 1 to 20.

14. L has the following structure: 【Chemistry 10】 The antibody-drug conjugate of any one of claims 1 to 10, selected from the group consisting of:

15. L has the following structure: 【Chemistry 11】 The antibody-drug conjugate of any one of claims 1 to 10, selected from the group consisting of:

16. E is a single bond, —NH—CH ２ --NH-CH ２ -O-CH ２ -CO-, 【Chemistry 12】 The antibody-drug conjugate according to any one of claims 1 to 15,

17. E is a single bond, NH—CH ２ --NH-CH ２ -O-CH ２ -CO-, 【Chemistry 13】 The antibody-drug conjugate according to any one of claims 1 to 15,

18. E is NH-CH ２ -, 【Chemistry 14】 The antibody-drug conjugate according to any one of claims 1 to 10, wherein

19. E is -NH-CH ２ -or 【Chemistry 15】 The antibody-drug conjugate according to any one of claims 1 to 10, wherein

20. 【Catalog 16】 but has the following structure: 【Chemistry 17】 【Chemistry 18】 【Chemistry 19】 【Chemistry 20】 The antibody-drug conjugate according to any one of claims 1 to 19, wherein the antibody-drug conjugate is selected from the group consisting of:

21. The antibody-drug conjugate of any one of claims 1 to 20, wherein the cytotoxic drug is selected from a tubulin inhibitor, a DNA intercalator, a DNA topoisomerase inhibitor, and an RNA polymerase inhibitor, or a pharmaceutically acceptable salt, ester, or analog thereof.

22. The antibody-drug conjugate of claim 21 , wherein the tubulin inhibitor is an auristatin compound or a maytansinoid compound.

23. The antibody-drug conjugate of claim 21, wherein the DNA intercalator is a pyrrolobenzodiazepine (PBD).

24. 22. The antibody-drug conjugate of claim 21, wherein the DNA topoisomerase inhibitor is a topoisomerase I inhibitor or a topoisomerase II inhibitor.

25. 25. The antibody-drug conjugate of claim 24, wherein the topoisomerase I inhibitor is camptothecin, hydroxycamptothecin, 9-aminocamptothecin, SN-38, irinotecan, topotecan, belotecan, or rubitecan.

26. 25. The antibody-drug conjugate of claim 24, wherein the topoisomerase II inhibitor is doxorubicin, PNU-159682, duocarmycin, daunorubicin, mitoxantrone, podophyllotoxin, or etoposide.

27. 22. The antibody-drug conjugate of claim 21, wherein the RNA polymerase inhibitor is α-amanitin, or a pharmaceutically acceptable salt, ester, or analog thereof.

28. the cytotoxic agent is 【Chemical 21】 The antibody-drug conjugate of claim 21,

29. the cytotoxic agent is selected from the compounds shown as Formula I and II; 【Chemical 22】 R １ and R ２ However, each independently, C １～６ selected from alkyl and halogen; R ３ is H and —CO—CH ２ OH, R ４ and R ５ are each independently selected from H, halogen, and hydroxyl, or R ４ and R ５ are connected to associated carbon atoms to form a 5- to 6-membered oxygen-containing heterocycle; R ６ is hydrogen and -C １～４ Alkylene -NR ａ R ｂ is selected from R ７ But hydrogen, C １～６ Alkyl, and -C １～４ Alkylene -NR ａ R ｂ is selected from In each occurrence, R ａ and R ｂ are each independently H, C １～６ Alkyl, —SO ２ -C １～６ Alkyl, and —CO—C １～６ The antibody-drug conjugate of claim 21, wherein the aryl group is selected from alkyl.

30. The cytotoxic agent is selected from the group consisting of the following compounds: 【Chemical 23】 is selected from The antibody-drug conjugate of claim 21, wherein the corresponding fragment of the cytotoxic drug obtained after the cytotoxic drug is linked to the linker is D as shown in the general formula:

31. D is -OH, -NH on the cytotoxic drug ２ or a monovalent structure obtained by losing one H from the secondary amine group.

32. Selected from ADC A-01 to ADC A-26, ADC B-01 to ADC B-06, and ADC C01 shown below; 【Chemistry 24】 【Chemistry 25】 【Chemical 26】 【Chemical 27】 【Chemical Formula 28】 【Chemical 29】 【Chemistry 30】 【Chemical 31】 【Chemical 32】 【Chemical 33】 HA in each antibody-drug conjugate represents an antibody or antigen-binding fragment thereof comprising a VH set forth as SEQ ID NO: 1, 3, 5, 7, or 9 and a VL set forth as SEQ ID NO: 2, 4, 6, 8, or 10; and 【Chemical 34】 32. The antibody-drug conjugate of claim 1, wherein represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and the linker.

33. the antibody-drug conjugate comprising the formula: 【Chemistry 35】 The HA in each antibody-drug conjugate is (1) An antibody or antigen-binding fragment thereof, comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a VL having the amino acid sequence set forth in SEQ ID NO: 2; (2) An antibody or antigen-binding fragment thereof, comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a VL having the amino acid sequence set forth in SEQ ID NO: 4; (3) An antibody or antigen-binding fragment thereof, comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a VL having the amino acid sequence set forth in SEQ ID NO: 6; (4) An antibody or antigen-binding fragment thereof, comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a VL having the amino acid sequence set forth in SEQ ID NO: 8; (5) An antibody or antigen-binding fragment thereof, comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a VL having the amino acid sequence set forth in SEQ ID NO: 10; 【Chemical 36】 represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and the linker; 33. The antibody-drug conjugate of any one of claims 1 to 32, wherein HA is linked via the sulfhydryl in the antibody or antigen-binding fragment to form the antibody-drug conjugate.

34. the antibody-drug conjugate comprising the formula: 【Chemical 37】 【Chemical 38】 The HA in each antibody-drug conjugate is (1) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (2) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (3) An antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (4) An antibody or antigen-binding fragment thereof comprising: a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45; and a VL having the amino acid sequence set forth in SEQ ID NO: 8 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; (5) An antibody or antigen-binding fragment thereof comprising: a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a CH having the amino acid sequence set forth in SEQ ID NO: 43 or 45; and a VL having the amino acid sequence set forth in SEQ ID NO: 10 and a CL having the amino acid sequence set forth in SEQ ID NO: 44; 【Chemical Formula 39】 represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and the linker, wherein: HA is linked via one or more sulfhydryls in the antibody or antigen-binding fragment to form the antibody-drug conjugate; The antibody-drug conjugate of any one of claims 1 to 32, wherein x is 3 to 9.

35. The antibody-drug conjugate of any one of claims 1 to 34, wherein the antibody or antigen-binding fragment thereof comprises a VH shown as SEQ ID NO: 3 and a CH shown as SEQ ID NO: 43 or 45, a VL shown as SEQ ID NO: 4 and a CL shown as SEQ ID NO:

44.

36. the antibody-drug conjugate is: 【Chemistry 40】 the HA in the antibody-drug conjugate is an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44; 【Chemistry 41】 represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and the linker; HA is linked via the sulfhydryl in the antibody or antigen-binding fragment to form the antibody-drug conjugate; The antibody-drug conjugate of any one of claims 1 to 33, wherein x is 7 to 9.

37. the antibody-drug conjugate is: 【Chemistry 42】 HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47; 【Chemistry 43】 represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and the linker; HA is linked via the sulfhydryl of the cysteine ​​in the antibody or antigen-binding fragment to form the antibody-drug conjugate; The antibody-drug conjugate of any one of claims 1 to 33, wherein x is 7 to 8.

38. the antibody-drug conjugate is: 【Chemical 44】 HA is an antibody comprising a heavy chain (HC) consisting of the amino acid sequence set forth in SEQ ID NO: 52, 53, or 54 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47; 【Chemistry 45】 represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and the linker; HA is linked via the sulfhydryl of the cysteine ​​in the antibody or antigen-binding fragment to form the antibody-drug conjugate; The antibody-drug conjugate of any one of claims 1 to 33, wherein x is 7 to 8.

39. the antibody-drug conjugate is: 【Chemistry 46】 HA is an antibody consisting of two heavy chains and two light chains, each heavy chain (HC) consisting of the amino acid sequence set forth in SEQ ID NO: 52, and each light chain (LC) consisting of the amino acid sequence set forth in SEQ ID NO: 47; 【Chemistry 47】 represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and the linker; HA is linked via the sulfhydryl of the cysteine ​​in the antibody or antigen-binding fragment to form the antibody-drug conjugate; The antibody-drug conjugate of any one of claims 1 to 33, wherein x is 7 to 8.

40. the antibody-drug conjugate is: 【Chemistry 48】 HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47; 【Chemistry 49】 represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and the linker; HA is linked via the sulfhydryl of the cysteine ​​in the antibody or antigen-binding fragment to form the antibody-drug conjugate; The antibody-drug conjugate of any one of claims 1 to 33, wherein x is 8.

41. the antibody-drug conjugate is: 【Chemistry 50】 HA is an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44; 【Chemistry 51】 represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and the linker; HA is linked via the sulfhydryl in the antibody or antigen-binding fragment to form the antibody-drug conjugate; The antibody-drug conjugate of any one of claims 1 to 33, wherein x is 7 to 9.

42. the antibody-drug conjugate is: 【Chemistry 52】 HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47; 【Chemistry 53】 represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and the linker; HA is linked via the sulfhydryl in the antibody or antigen-binding fragment to form the antibody-drug conjugate; The antibody-drug conjugate of any one of claims 1 to 33, wherein x is 7 to 8.

43. the antibody-drug conjugate is: 【Chemical 54】 HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47; 【Chemistry 55】 represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and the linker; HA is linked via the sulfhydryl in the antibody or antigen-binding fragment to form the antibody-drug conjugate; The antibody-drug conjugate of any one of claims 1 to 33, wherein x is 8.

44. the antibody-drug conjugate is: 【Chemical Formula 56】 HA is an antibody or antigen-binding fragment thereof comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44; 【Chemical 57】 represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and the linker; HA is linked via the sulfhydryl in the antibody or antigen-binding fragment to form the antibody-drug conjugate; The antibody-drug conjugate of any one of claims 1 to 33, wherein x is 7 to 9.

45. the antibody-drug conjugate is: 【Chemistry 58】 HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47; 【Chemical 59】 represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and the linker; HA is linked via the sulfhydryl in the antibody or antigen-binding fragment to form the antibody-drug conjugate; The antibody-drug conjugate of any one of claims 1 to 33, wherein x is 7 to 8.

46. the antibody-drug conjugate is: 【Chemistry 60】 HA is an antibody or antigen-binding fragment thereof comprising a heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46 and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47; 【Hua 61】 represents the linkage formed from a sulfhydryl in the antibody or antigen-binding fragment and the linker; HA is linked via the sulfhydryl in the antibody or antigen-binding fragment to form the antibody-drug conjugate; The antibody-drug conjugate of any one of claims 1 to 33, wherein x is 8.

47. 34. A composition comprising one or more antibody-drug conjugates of any one of claims 1-33, wherein the DAR (drug-antibody conjugate ratio) of the composition is 1-2, 1-3, 1-4, 1-5, 1-6, 1-7, 1-8, 1-9, 1-10, 2-3, 2-4, 2-5, 2-6, 2-7, 2-8, 2-9, 2-10, 3-4, 3-5, 3-6, 3-7, 3-8, 3-9, 3-10, 4-5, 4-6, 4-7, 4-8, 4-9, 4-10, 5-6, 5-7, 5-8, 5-9, 5-10, 6-7, 6-8, 6-9, 6-10, 7-8, 7-9, 7-10, 8-9, 8-10, or 9-10.

48. The antibody-drug conjugate of any one of claims 1 to 47, wherein (i) the heavy chain C-terminus lacks a lysine residue, (ii) the heavy chain N-terminus is glutamine, glutamic acid, pyroglutamate, or pyroglutamic acid, or (iii) the heavy chain C-terminus lacks a lysine residue and the heavy chain N-terminus is glutamine, glutamic acid, pyroglutamate, or pyroglutamic acid.

49. The antibody-drug conjugate of any one of claims 1 to 46 and 48, wherein the antibody or antigen-binding fragment comprises a heavy chain comprising a VH having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, and 9.

50. The antibody-drug conjugate of any one of claims 1 to 46, 48, and 49, wherein the antibody or antigen-binding fragment comprises a heavy chain comprising a VH having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, and 9, and the N-terminal glutamine or glutamic acid of the antibody variable region is cyclized to pyroglutamate or pyroglutamic acid.

51. The antibody-drug conjugate of any one of claims 1 to 46 and 48 to 50, wherein the antibody or antigen-binding fragment comprises a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 8, and the N-terminal glutamic acid of the antibody light chain variable region is cyclized to pyroglutamate or pyroglutamic acid.

52. the antibody or antigen-binding fragment: (a) a heavy chain consisting of the sequence set forth as SEQ ID NO: 52 and a light chain consisting of the sequence set forth as SEQ ID NO: 47; (b) a heavy chain consisting of the sequence set forth as SEQ ID NO: 53 and a light chain consisting of the sequence set forth as SEQ ID NO: 47; or (c) an antibody-drug conjugate according to any one of claims 1 to 46 and 48 to 51, comprising a heavy chain consisting of the sequence shown as SEQ ID NO: 54 and a light chain consisting of the sequence shown as SEQ ID NO:

47.

53. The antibody or antigen-binding fragment thereof can be obtained by expressing in a host cell nucleic acid molecules encoding the heavy chain and the light chain of the antibody or antigen-binding fragment thereof, and the antibody or antigen-binding fragment thereof comprises: (1) A heavy chain comprising a VH sequence shown as SEQ ID NO: 1 and a heavy chain constant region (CH) shown as SEQ ID NO: 43, and a light chain comprising a VL sequence shown as SEQ ID NO: 2 and a light chain constant region (CL) described in SEQ ID NO: 44; (2) A heavy chain comprising a VH sequence shown as SEQ ID NO: 3 and a heavy chain constant region (CH) shown as SEQ ID NO: 43 or 45, and a light chain comprising a VL sequence shown as SEQ ID NO: 4 and a light chain constant region (CL) described in SEQ ID NO: 44; (3) A heavy chain comprising a VH sequence shown as SEQ ID NO: 5 and a heavy chain constant region (CH) shown as SEQ ID NO: 43, and a light chain comprising a VL sequence shown as SEQ ID NO: 6 and a light chain constant region (CL) described in SEQ ID NO: 44; (4) A heavy chain comprising a VH sequence shown as SEQ ID NO: 7 and a heavy chain constant region (CH) shown as SEQ ID NO: 43, and a light chain comprising a VL sequence shown as SEQ ID NO: 8 and a light chain constant region (CL) described in SEQ ID NO: 44; (5) A heavy chain comprising a VH sequence shown as SEQ ID NO: 9 and a heavy chain constant region (CH) shown as SEQ ID NO: 43, and a light chain comprising a VL sequence shown as SEQ ID NO: 10 and a light chain constant region (CL) described in SEQ ID NO: 44; (6) A heavy chain (HC) having the amino acid sequence set forth in SEQ ID NO: 46, and a light chain (LC) having the amino acid sequence set forth in SEQ ID NO: 47; (7) VH having the amino acid sequence set forth in SEQ ID NO: 1, and VL having the amino acid sequence set forth in SEQ ID NO: 2; (8) VH having the amino acid sequence set forth in SEQ ID NO: 3, and VL having the amino acid sequence set forth in SEQ ID NO: 4; (9) VH having the amino acid sequence set forth in SEQ ID NO: 5, and VL having the amino acid sequence set forth in SEQ ID NO: 6; (10) VH having the amino acid sequence set forth in SEQ ID NO: 7 and VL having the amino acid sequence set forth in SEQ ID NO: 8, or (11) The antibody-drug conjugate according to any one of claims 1 to 46 and 48 to 52, comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a VL having the amino acid sequence set forth in SEQ ID NO:

10.

54. An antibody-drug conjugate comprising an antibody, wherein the antibody binds to PTK7 linked via one or more cysteine ​​or lysine residues or non-standard amino acid substitutions of amino acids of the antibody to a drug linker having a structure shown as formula M-LE-D; M, 【Hua 62】 wherein ring A is a 5- to 6-membered aliphatic heterocycle or a 5- to 20-membered aromatic ring system, and the aliphatic heterocycle and aromatic ring system are selected from the group consisting of oxo (=O), halogen, cyano, amino, carboxyl, mercapto, and C １～６ optionally substituted with one or more groups selected from the group consisting of alkyl; M １ is a single bond, C １～２０ Alkylene, C ２～２０ Alkenylene, and C ２～２０ alkynylene, L is a linker between M and E; E is a structural fragment connecting L and D; An antibody-drug conjugate wherein D is a cytotoxic drug fragment.

55.

63. but, 【Hua 64】 【Chemistry 65】 【Hua 66】 、 【Hua 67】 The antibody-drug conjugate of claim 54, selected from

56. The cytotoxic agent is selected from the group consisting of the following compounds: 【Chemistry 68】 is selected from 56. The antibody-drug conjugate of claim 54 or 55, wherein the corresponding fragment of the cytotoxic drug obtained after the cytotoxic drug is linked to the linker is D as shown in the general formula:

57. The antibody that binds to PTK7 is (1) A heavy chain variable region (VH) and / or light chain variable region (VL) of the following, wherein the CDRs are defined according to the Chothia numbering system: (1a) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 11 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 12 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof; or (1b) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 27 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 28 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof; The variant of any of (1a) and (1b) has a heavy chain variable region (VH) and / or a light chain variable region (VL) that has at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence from which the variant is derived, or that has one or several amino acid substitutions, deletions, or additions with the sequence from which the variant is derived; or (2) The following heavy chain variable region (VH) and / or light chain variable region (VL): (2a) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence of SEQ ID NO: 17 or a variant thereof, CDR-H2 having the amino acid sequence of SEQ ID NO: 18 or 19 or a variant thereof, CDR-H3 having the amino acid sequence of SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence of SEQ ID NO: 14 or a variant thereof, CDR-L2 having the amino acid sequence of SEQ ID NO: 15 or a variant thereof, CDR-L3 having the amino acid sequence of SEQ ID NO: 16 or a variant thereof, or (2b) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 33 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 34 or 35 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof; The variant of any of (2a) and (2b) has a heavy chain variable region (VH) and / or a light chain variable region (VL) that has at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence from which the variant is derived, or that has one or several amino acid substitutions, deletions, or additions with the sequence from which the variant is derived; or (3) The following heavy chain variable region (VH) and / or light chain variable region (VL): wherein the CDRs are defined according to the IMGT numbering system: (3a) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 20 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 21 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 22 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 23 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 24 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof; or (3b) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 36 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 37 or a variant thereof, CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 38 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 39 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 40 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof; The variants of any of (3a) and (3b) have at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence from which the variants are derived, or have one or several amino acid substitutions, deletions, or additions with the sequence from which the variants are derived; and (4) The following heavy chain variable region (VH) and / or light chain variable region (VL), wherein the CDRs are defined according to the AbM numbering system: (4a) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 25 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 26 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 13 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 14 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 15 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 16 or a variant thereof; or (4b) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the amino acid sequence set forth in SEQ ID NO: 41 or a variant thereof, CDR-H2 having the amino acid sequence set forth in SEQ ID NO: 42 or a variant thereof, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO: 29 or a variant thereof, and / or a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 30 or a variant thereof, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 31 or a variant thereof, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof; The antibody-drug conjugate of claim 54 or 55, wherein the variant of either (4a) or (4b) is selected from the group consisting of a heavy chain variable region (VH) and / or a light chain variable region (VL) that has at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity compared to the sequence from which the variant is derived, or that has one or several amino acid substitutions, deletions, or additions compared to the sequence from which the variant is derived.

58. The antibody that binds to PTK7 is (a) a VH having the amino acid sequence set forth in SEQ ID NO: 1 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 2 or a variant thereof; (b) a VH having the amino acid sequence set forth in SEQ ID NO: 3 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 4 or a variant thereof; (c) a VH having the amino acid sequence set forth in SEQ ID NO: 5 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 6 or a variant thereof; (d) a VH having the amino acid sequence set forth in SEQ ID NO: 7 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 8 or a variant thereof; and (c) a VH having the amino acid sequence set forth in SEQ ID NO: 9 or a variant thereof, and / or a VL having the amino acid sequence set forth in SEQ ID NO: 10 or a variant thereof; The antibody-drug conjugate of claim 57, wherein the variant has at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity compared to the sequence from which the variant is derived, or has one or several amino acid substitutions, deletions, or additions compared to the sequence from which the variant is derived.

59. The antibody that binds to PTK7 is (1) A heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 1 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43, and a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 2 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44; (2) A heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 3 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43 or 45, and a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 4 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44; (3) A heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 5 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43, and a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 6 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44; (4) A heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 7 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43, and a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 8 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO: 44; (5) The antibody-drug conjugate of claim 54 or 55, which is selected from the group consisting of a heavy chain comprising a VH having the amino acid sequence set forth in SEQ ID NO: 9 and a heavy chain constant region (CH) having the amino acid sequence set forth in SEQ ID NO: 43, and a light chain comprising a VL having the amino acid sequence set forth in SEQ ID NO: 10 and a light chain constant region (CL) having the amino acid sequence set forth in SEQ ID NO:

44.

60. 56. The antibody-drug conjugate of claim 54 or 55, wherein the antibody that binds to PTK7 comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO:46 and a light chain having the amino acid sequence set forth in SEQ ID NO:

47.

61. A pharmaceutical composition comprising the antibody-drug conjugate of any one of claims 1 to 60, and one or more pharmaceutically acceptable excipients.

62. 62. A method for treating cancer in a subject with high expression of PTK7, comprising administering to the subject a therapeutically effective amount of the antibody-drug conjugate of any one of claims 1 to 60, or the pharmaceutical composition of claim 61.

63. 63. The method of claim 62, wherein the cancer comprises a solid tumor or a hematological malignancy.

64. 64. The method of claim 63, wherein the cancer is lung cancer, breast cancer, epidermal cancer, ovarian cancer, or esophageal cancer.

65. Use of the antibody-drug conjugate of any one of claims 1 to 60 or the pharmaceutical composition of claim 61 in the manufacture of a drug for treating cancer associated with high expression of PTK7.

66. 66. The use of claim 65, wherein the cancer comprises a solid tumor or a hematological malignancy.

67. 66. The use of claim 65, wherein the cancer associated with high expression of PTK7 is lung cancer, breast cancer, epidermal cancer, ovarian cancer, and esophageal cancer.

68. Use of the antibody-drug conjugate of any one of claims 1 to 60 or the pharmaceutical composition of claim 61 in the treatment of cancer associated with high expression of PTK7.

69. 69. The use of claim 68, wherein the cancer comprises a solid tumor or a hematological malignancy.

70. 70. The use of claim 69, wherein the cancer associated with high expression of PTK7 is lung cancer, breast cancer, epidermal cancer, ovarian cancer, and esophageal cancer.

71. The antibody-drug conjugate of any one of claims 1 to 60 or the pharmaceutical composition of claim 61 for the treatment of cancer associated with high expression of PTK7.

72. 72. The antibody-drug conjugate of claim 71, wherein the cancer comprises a solid tumor or a hematological malignancy.

73. The antibody-drug conjugate of claim 72, wherein the cancer is lung cancer, breast cancer, epidermal cancer, or esophageal cancer.

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