ADAM9 binding protein and its uses

Abstract

The present invention relates to the field of disease treatment. The present invention provides an anti-ADAM9 antibody or its antigen-binding fragment, a nucleic acid molecule encoding the same, and a method for preparing the same. The anti-ADAM9 antibody or its antigen-binding fragment has high specificity and high affinity for ADAM9. The present invention also relates to the use of the antibody or its antigen-binding fragment in the treatment and diagnosis of diseases.

Description

Technology Field

[0001] Cross-reference regarding related applications

[0002] This application claims priority to Chinese patent application No. 202311734757.9 filed on December 14, 2023, Chinese patent application No. 202410173238.8 filed on February 6, 2024, and Chinese patent application No. 202411789351.5 filed on December 6, 2024, the contents of which are incorporated herein by reference in their entirety.

[0003] Technology field

[0004] The present disclosure relates to the field of therapeutic monoclonal antibodies. Specifically, the present disclosure relates to antibodies against ADAM9. The present disclosure also relates to the use of said antibodies in the treatment and diagnosis of diseases. Background Technology

[0005] ADAMs (a disintegrin and a metalloprotease) are type I transmembrane proteins anchored to the cell surface membrane. More than 30 types have been discovered to date. Similar to matrix metalloproteinases (MMPs), ADAMs also contain a prodomain and a zinc-binding metalloproteinase domain. Additionally, ADAMs possess a disintegrin domain unique among cell surface proteins. ADAM9 is widely expressed in the human body, regulates various biological functions, and plays an important role in a wide range of diseases, including neurodegenerative diseases, retinal diseases, inflammation, and tumors. ADAM9 is involved in various physiological functions primarily through a protein-degrading domain involved in adhesion and a metalloprotein domain involved in the cleavage and shedding of various cell surface proteins.

[0006] Recently, there has been increasing evidence that ADAM9 plays a significant role in tumors. ADAM9 is involved in the regulation of various tumor processes. In addition to metastasis, ADAM9 plays a crucial role in tumor proliferation, angiogenesis, and even immune evasion. Overexpression of ADAM9 has been found in various cancers, and this is associated with tumor invasiveness and poor prognosis. Furthermore, ADAM9 promotes tumor progression, treatment resistance, and cancer metastasis through proteolytic or non-proteolytic pathways.

[0007] Many studies have revealed that ADAM9 overexpression shortens overall survival. Further research has shown that ADAM9 plays a major role in various stages of lung cancer metastasis. Shintani et al. were the first to report that ADAM9 overexpression promotes the adhesion of tumor cells to vascular endothelial cells, suggesting the importance of ADAM9 in the metastatic process. Furthermore, ADAM9 promotes metastasis by enhancing cell migration and stress resistance through novel mechanisms. According to a study by Fritzsche et al., the overexpression of ADAM9 mRNA and protein was found to be associated with low recurrence-free survival in prostate cancer. Immunohistochemical analysis revealed increased ADAM9 protein expression in more than 60% of recurrent prostate tumors. IL-6 is a key mediator of liver cancer invasion and metastasis, and IL-6 promotes ADAM9 expression in vitro by activating the JNK pathway. Silencing ADAM9 can not only reduce the size of primary tumors but also suppress the rate of metastasis to the lungs. Conversely, overexpression of ADAM9 accelerates the growth of primary tumors and promotes metastasis to the lungs. ADAM9 shares methyltransferase with EGFR. Nuclear receptor-binding SET domain protein 2 is a member of the histone methyltransferase family and can promote TNBC cell resistance to EGFR inhibitors by upregulating the expression of ADAM9 and EGFR. KRAS signaling is essential for maintaining oncogenesis in pancreatic cancer. Yuan et al. discovered that dysregulation of KRAS signaling promotes ADAM9 expression via the NF-kB cascade, and that ADAM9 knockout inhibits downstream pathways of KRAS and MEK-ERK signaling.In addition, studies have revealed that circular ADAM9 (circ-ADAM9) is upregulated in pancreatic cancer cells, which is associated with a poor prognosis. Overexpression of circ-ADAM9 increases ERK signaling and promotes cell proliferation and migration in vitro, whereas silencing of circ-ADAM9 delays the growth of pancreatic tumors in vivo.

[0008] Studies conducted to date provide sufficient support for the use of ADAM9 as a prognostic marker for cancer. Currently used therapeutic drugs (e.g., sorafenib and regorafenib) enhance cancer treatment efficacy by reducing ADAM9 levels, offering new ideas for ADAM9-targeted therapy. At the 2021 AACR conference, ImmunoGen presented the progress of the IMGC936 study. IMGC936 is the first ADAM9-targeted antibody-drug conjugate (ADC) co-developed by ImmunoGen and Macrogenics. Composed of a high-affinity humanized monoclonal antibody, a mytansinoid microtubule inhibitor payload, and a stable tripeptide linker, IMGC936 is an innovative drug with a drug-to-antibody ratio (DAR) of 2 and is currently in Phase I clinical trials.

[0009] In terms of safety, no major developmental defects were observed in ADAM9 gene-knockout mice, except for subsequent retinal defects. This suggests that it may be a relatively safe target for adult patients.

[0010] Nevertheless, when treating ADAM9-related diseases, the specificity of therapeutic agents against ADAM9 must be considered, as regulating the protective activity of MMPs generally leads to side effects. Therefore, it is necessary to develop antibodies targeting ADAM9, which will provide patients with a wider range of treatment options. The problem to be solved

[0011] Summary of the Invention

[0012] In the present application, the inventors have developed a high-affinity humanized antibody with excellent properties that can specifically recognize / bind to ADAM9 and can be used for the prevention and / or treatment of cancers expressing ADAM9, specifically cancers overexpressing ADAM9. means of solving the problem

[0013] Antibody of the present invention

[0014] In one aspect, the present disclosure provides an antibody or its antigen-binding fragment that specifically binds to ADAM9, and said antibody or its antigen-binding fragment comprises complementarity determining regions (CDRs) as follows:

[0015] (a) CDR-H1, CDR-H2, and CDR-H3 contained in the heavy chain variable region (VH) presented in SEQ ID NO: 54 or 52; and / or CDR-L1, CDR-L2, and CDR-L3 contained in the light chain variable region (VL) presented in SEQ ID NO: 55 or 53;

[0016] (b) CDR-H1, CDR-H2, and CDR-H3 contained in the heavy chain variable region (VH) presented in SEQ ID NO: 3 or 1; and / or CDR-L1, CDR-L2, and CDR-L3 contained in the light chain variable region (VL) presented in SEQ ID NO: 4 or 2;

[0017] (c) CDR-H1, CDR-H2, and CDR-H3 contained in the heavy chain variable region (VH) presented in SEQ ID NO: 9 or 7; and / or CDR-L1, CDR-L2, and CDR-L3 contained in the light chain variable region (VL) presented in SEQ ID NO: 10 or 8;

[0018] (d) CDR-H1, CDR-H2, and CDR-H3 contained in the heavy chain variable region (VH) presented in SEQ ID NO: 50 or 48; and / or CDR-L1, CDR-L2, and CDR-L3 contained in the light chain variable region (VL) presented in SEQ ID NO: 51 or 49; or

[0019] (e) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) described above, and / or CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) described above, wherein the heavy chain variable region (VH) and / or light chain variable region (VL) comprises a mutation in at least one CDR compared to any one of the heavy chain variable region and / or light chain variable region (a) to (d), said mutation being a substitution, deletion, or addition of one or several amino acids (e.g., a substitution, deletion, or addition of 1, 2, or 3 amino acids).

[0020] In certain embodiments, the antibody or its antigen-binding fragment comprises complementarity determining regions (CDRs) as follows:

[0021] (i) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) presented in SEQ ID NO: 54; and / or CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) presented in SEQ ID NO: 55;

[0022] (ii) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) presented in SEQ ID NO: 52; and / or CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) presented in SEQ ID NO: 53;

[0023] (iii) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) presented in SEQ ID NO: 3; and / or CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) presented in SEQ ID NO: 4;

[0024] (iv) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) presented in SEQ ID NO: 1; and / or CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) presented in SEQ ID NO: 2;

[0025] (v) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) presented in SEQ ID NO: 9; and / or CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) presented in SEQ ID NO: 10;

[0026] (vi) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) presented in SEQ ID NO: 7; and / or CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) presented in SEQ ID NO: 8;

[0027] (vii) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) presented in SEQ ID NO: 50; and / or CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) presented in SEQ ID NO: 51;

[0028] (viii) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) presented in SEQ ID NO: 48; and / or CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) presented in SEQ ID NO: 49; or

[0029] (ix) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) described above, and / or CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) described above, wherein the heavy chain variable region (VH) and / or light chain variable region (VL) comprises a mutation in at least one CDR compared to any one of the heavy chain variable region and / or light chain variable region (i) or (iv), said mutation being a substitution, deletion, or addition of one or several amino acids (e.g., a substitution, deletion, or addition of 1, 2, or 3 amino acids).

[0030] In certain embodiments, the substitution is a conservative substitution.

[0031] In certain embodiments, the CDRs are defined according to the IMGT, Kabat, Chothia, or AbM numbering system.

[0032] In certain embodiments, the ADAM9 comprises human ADAM9 and / or monkey ADAM9. In certain embodiments, the monkey is a Macaca mulatta.

[0033] In certain embodiments, the antibody of the present disclosure or its antigen-binding fragment comprises the following heavy chain variable region (VH) and / or light chain variable region (VL), where CDRs are defined by the IMGT numbering system:

[0034] (1a) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 81 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 82 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 83 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 84 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 85 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 76 or a variant thereof; and

[0035] (1b-i) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 21 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 22 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 23 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 24 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 25 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 16 or a variant thereof; and

[0036] (1b-ii) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 21 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 22 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 94 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 24 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 25 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 16 or a variant thereof; and

[0037] (1c-i) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 35 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 36 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 37 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 38 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 39 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 30 or a variant thereof; and

[0038] (1c-ii) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 103 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 104 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 37 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 105 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 39 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 30 or a variant thereof; or

[0039] (1d) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 65 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 66 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 67 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 68 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 69 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 70 or a variant thereof;

[0040] A variant described in any one of (1a), (1b-i), (1b-ii), (1c-i), (1c-ii) and (1d) has one or more amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) compared to the sequence from which the variant is derived. In certain embodiments, the substitutions are conservative substitutions.

[0041] In certain embodiments, the antibody of the present disclosure or its antigen-binding fragment comprises the following heavy chain variable region (VH) and / or light chain variable region (VL), as defined by the Chothia numbering system of CDRs:

[0042] (2a) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 71 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 72 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 73 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 74 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 75 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 76 or a variant thereof; and

[0043] (2b) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 11 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 12 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 13 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 14 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 15 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 16 or a variant thereof; and

[0044] (2c-i) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 11 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 26 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 27 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 28 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 29 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 30 or a variant thereof;

[0045] (2c-ii) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 95 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 96 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 27 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 97 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 98 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 30 or a variant thereof; or

[0046] (2d) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 56 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 57 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 58 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 59 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 60 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 61 or a variant thereof;

[0047] A variant described in any one of (2a), (2b), (2c-i), (2c-ii) and (2d) has one or more amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) compared to the sequence from which the variant is derived. In certain embodiments, the substitutions are conservative substitutions.

[0048] In certain embodiments, the antibody of the present disclosure or its antigen-binding fragment comprises the following heavy chain variable region (VH) and / or light chain variable region (VL), as defined by the Kabat numbering system of CDRs:

[0049] (3a) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 79 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 80 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 73 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 74 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 75 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 76 or a variant thereof;

[0050] (3b) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 19 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 20 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 13 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 14 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 15 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 16 or a variant thereof;

[0051] (3c-i) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 33 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 34 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 27 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 28 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 29 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 30 or a variant thereof;

[0052] (3c-ii) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 101 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 102 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 27 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 97 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 98 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 30 or a variant thereof; or

[0053] (3d) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 62 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 63 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 64 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 59 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 60 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 61 or a variant thereof;

[0054] A variant described in any one of (3a), (3b), (3c-i), (3c-ii) and (3d) has one or more amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) compared to the sequence from which the variant is derived. In certain embodiments, the substitutions are conservative substitutions.

[0055] In certain embodiments, the antibody of the present disclosure or its antigen-binding fragment comprises the following heavy chain variable region (VH) and / or light chain variable region (VL), where CDRs are defined by the AbM numbering system:

[0056] (4a) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 77 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 78 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 73 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 74 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 75 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 76 or a variant thereof;

[0057] (4b) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 17 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 18 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 13 or a variant thereof; and / or, light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 14 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 15 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 16 or a variant thereof;

[0058] (4c-i) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 31 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 32 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 27 or a variant thereof; and / or, light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 28 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 29 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 30 or a variant thereof;

[0059] (4c-ii) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 99 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 100 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 27 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 97 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 98 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 30 or a variant thereof; or

[0060] (4d) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 62 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 63 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 64 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 59 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 60 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 61 or a variant thereof;

[0061] A variant described in any one of (4a), (4b), (4c-i), (4c-ii) and (4d) has one or more amino acid substitutions, deletions, or additions (e.g., one, two, or three amino acid substitutions, deletions, or additions) compared to the sequence from which the variant is derived. In certain embodiments, the substitutions are conservative substitutions.

[0062] In certain embodiments, the antibody or its antigen-binding fragment of the present disclosure comprises:

[0063] (ai) VH comprising the sequence presented in SEQ ID NO: 54 or a variant thereof and / or VL comprising the sequence presented in SEQ ID NO: 55 or a variant thereof;

[0064] (a-ii) VH comprising the sequence presented in SEQ ID NO: 52 or a variant thereof and / or VL comprising the sequence presented in SEQ ID NO: 53 or a variant thereof;

[0065] (bi) VH comprising the sequence presented in SEQ ID NO: 3 or a variant thereof and / or VL comprising the sequence presented in SEQ ID NO: 4 or a variant thereof;

[0066] (b-ii) VH comprising the sequence presented in SEQ ID NO: 1 or a variant thereof and / or VL comprising the sequence presented in SEQ ID NO: 2 or a variant thereof;

[0067] (ci) VH comprising the sequence presented in SEQ ID NO: 9 or a variant thereof and / or VL comprising the sequence presented in SEQ ID NO: 10 or a variant thereof;

[0068] (c-ii) VH comprising the sequence presented in SEQ ID NO: 7 or a variant thereof and / or VL comprising the sequence presented in SEQ ID NO: 8 or a variant thereof;

[0069] (di) VH comprising the sequence presented in SEQ ID NO: 50 or a variant thereof and / or VL comprising the sequence presented in SEQ ID NO: 51 or a variant thereof; or

[0070] (d-ii) VH comprising the sequence presented in SEQ ID NO: 48 or a variant thereof and / or VL comprising the sequence presented in SEQ ID NO: 49 or a variant thereof;

[0071] 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 with respect to the sequence from which the variant is derived, or has one or more amino acid substitutions, deletions, or additions (e.g., substitutions, deletions, or additions of 1, 2, 3, 4, or 5 amino acids) compared to the sequence from which the variant is derived; preferably, the substitutions are conservative substitutions.

[0072] In certain embodiments of the antibody or antigen-binding fragment disclosed herein, the heavy chain constant domain may include C-terminal lysine or lack C-terminal lysine or C-terminal glycine-lysine dipeptide. In some embodiments of the antibody or its antigen-binding fragment, the N-terminal amino acid of the antibody or its antigen-binding fragment may be cyclized with pyroglutamic acid. In some embodiments of the antibody or its antigen-binding fragment, the N-terminal amino acid of the antibody or its antigen-binding fragment may be cyclized with pyroglutamic acid.

[0073] As is known to those skilled in the art, pyroglutamic acid is the conjugate acid of pyroglutamate and is in equilibrium with pyroglutamate in solution.

[0074] In certain embodiments, the present invention provides a composition comprising the antibody or antigen-binding fragment disclosed herein, said antibody or antigen-binding fragment may independently comprise C-terminal lysine, lack C-terminal lysine, lack C-terminal glycine-lysine, and / or comprise N-terminal glutamine or glutamic acid, an N-terminal amino acid cyclized with pyroglutamic acid or an N-terminal amino acid cyclized with pyroglutamate.

[0075] In certain embodiments, the antibody or antigen-binding fragment disclosed herein comprises an antibody or antigen-binding fragment that specifically binds to an antigen and may include post-translational modifications thereof (e.g., cleavage of C-terminal lysine in the heavy chain, and conversion of N-terminal glutamine or glutamate in the heavy or light chain to pyroglutamate or pyroglutamate) that may occur during recombinant expression in a host cell (e.g., CHO cell) or during purification / storage.

[0076] In certain embodiments, the N-terminal glutamine of VH comprising the sequence presented in SEQ ID NO: 54, 52, 3, 1, 9, 7, 50, or 48 or a variant thereof undergoes a cyclization reaction to form pyroglutamic acid or pyroglutamate. In certain embodiments, the N-terminal glutamine of VH comprising the sequence presented in SEQ ID NO: 54, 3, 9, or 50 or a variant thereof undergoes a cyclization reaction to form pyroglutamic acid or pyroglutamate.

[0077] In certain embodiments, an antibody or its antigen-binding fragment having any one of features (1a), (2a), (3a), (4a), (ai) or (a-ii) in the embodiments further has a feature selected from the following:

[0078] (1) Less than about 100 ng / mL, e.g., less than about 80 ng / mL, less than 50 ng / mL, less than 20 ng / mL, less than 15 ng / mL, less than 14 ng / mL, less than 13 ng / mL, less than 12 ng / mL, less than 11 ng / mL, less than 10 ng / mL, less than 9 ng / mL, less than 8 ng / mL, less than 7 ng / mL, less than 6 ng / mL, less than 5 ng / mL, less than 4 ng / mL or less EC 50 , preferably EC measured by ELISA 50 As, combination with ADAM9 (e.g., human or monkey ADAM9);

[0079] (2) binding to ADAM9 (e.g., human or monkey ADAM9) with a KD of less than about 100 nM, e.g., less than about 90 nM, less than 80 nM, less than 70 nM, less than 60 nM, less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 15 nM, less than 10 nM, less than 5 nM or less than the KD, preferably with a KD measured by BLI (bio-layer interferometry) (e.g., ForteBio Octet®);

[0080] (3) For example, having CDC activity such as inducing apoptosis of ADAM9-expressing cells (e.g., tumor cells) through CDC;

[0081] (4) Does not have ADCC activity;

[0082] (5) Induction of ADAM9 internalization, for example, determined by flow cytometry;

[0083] (6) Does not bind to ADAM12 and ADAM15 proteins;

[0084] (7) Inhibition of cell proliferation (e.g., tumor cells); and / or

[0085] (8) Inhibition of tumor growth.

[0086] In certain embodiments, an antibody or its antigen-binding fragment having any one of the features of (1b-i), (1b-ii), (2b), (3b), (4b), (bi) or (b-ii) in the embodiments further has a feature selected from the following:

[0087] (1) Less than about 100 ng / mL, e.g., less than about 80 ng / mL, less than 50 ng / mL, less than 20 ng / mL, less than 15 ng / mL, less than 14 ng / mL, less than 13 ng / mL, less than 12 ng / mL, less than 11 ng / mL, less than 10 ng / mL, less than 9 ng / mL, less than 8 ng / mL, less than 7 ng / mL, less than 6 ng / mL, less than 5 ng / mL, less than 4 ng / mL or less EC 50 , preferably EC measured by ELISA 50 As, combination with ADAM9 (e.g., human or monkey ADAM9);

[0088] (2) binding to ADAM9 (e.g., human or monkey ADAM9) with a KD of less than about 100 nM, e.g., less than about 90 nM, less than 80 nM, less than 70 nM, less than 60 nM, less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 15 nM, less than 10 nM, less than 5 nM or less than the KD, preferably with a KD measured by BLI (bio-layer interferometry) (e.g., ForteBio Octet®);

[0089] (3) For example, having CDC activity such as inducing apoptosis of ADAM9-expressing cells (e.g., tumor cells) through CDC;

[0090] (4) Does not have ADCC activity;

[0091] (5) Induction of ADAM9 internalization, for example, determined by flow cytometry;

[0092] (6) Does not bind to ADAM12 and ADAM15 proteins;

[0093] (7) Inhibition of cell proliferation (e.g., tumor cells); and / or

[0094] (8) Inhibition of tumor growth.

[0095] In certain embodiments, an antibody or its antigen-binding fragment having any one of the features of (1c-i), (1c-ii), (2c-i), (2c-ii), (3c-i), (3c-ii), (4c-i), (4c-ii), (ci) or (c-ii) in the above embodiments further has a feature selected from the following:

[0096] (1) Less than about 100 ng / mL, e.g., less than about 80 ng / mL, less than 50 ng / mL, less than 20 ng / mL, less than 15 ng / mL, less than 14 ng / mL, less than 13 ng / mL, less than 12 ng / mL, less than 11 ng / mL, less than 10 ng / mL, less than 9 ng / mL, less than 8 ng / mL, less than 7 ng / mL, less than 6 ng / mL, less than 5 ng / mL, less than 4 ng / mL or less EC 50 , preferably EC measured by ELISA 50 As, combination with ADAM9 (e.g., human or monkey ADAM9);

[0097] (2) binding to ADAM9 (e.g., human or monkey ADAM9) with a KD of less than about 100 nM, e.g., less than about 90 nM, less than 80 nM, less than 70 nM, less than 60 nM, less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 15 nM, less than 10 nM, less than 5 nM or less than the KD, preferably with a KD measured by BLI (bio-layer interferometry) (e.g., ForteBio Octet®);

[0098] (3) For example, having CDC activity such as inducing apoptosis of ADAM9-expressing cells (e.g., tumor cells) through CDC;

[0099] (4) Does not have ADCC activity;

[0100] (5) Induction of ADAM9 internalization, for example, determined by flow cytometry;

[0101] (6) Does not bind to ADAM12 and ADAM15 proteins;

[0102] (7) Inhibition of cell proliferation (e.g., tumor cells); and / or

[0103] (8) Inhibition of tumor growth.

[0104] In certain embodiments, an antibody or its antigen-binding fragment having any one of features (1d), (2d), (3b), (3d), (4d), (di) or (d-ii) in the embodiments further has a feature selected from the following:

[0105] (1) Less than about 100 ng / mL, e.g., less than about 80 ng / mL, less than 50 ng / mL, less than 20 ng / mL, less than 15 ng / mL, less than 14 ng / mL, less than 13 ng / mL, less than 12 ng / mL, less than 11 ng / mL, less than 10 ng / mL, less than 9 ng / mL, less than 8 ng / mL, less than 7 ng / mL, less than 6 ng / mL, less than 5 ng / mL, less than 4 ng / mL or less EC 50 , preferably EC measured by ELISA 50 As, combination with ADAM9 (e.g., human or monkey ADAM9);

[0106] (2) binding to ADAM9 (e.g., human or monkey ADAM9) with a KD of less than about 100 nM, e.g., less than about 90 nM, less than 80 nM, less than 70 nM, less than 60 nM, less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 15 nM, less than 10 nM, less than 5 nM or less than the KD, preferably with a KD measured by BLI (bio-layer interferometry) (e.g., ForteBio Octet®);

[0107] (3) For example, having CDC activity such as inducing apoptosis of ADAM9-expressing cells (e.g., tumor cells) through CDC;

[0108] (4) Does not have ADCC activity;

[0109] (5) Induction of ADAM9 internalization, for example, determined by flow cytometry;

[0110] (6) Does not bind to ADAM12 and ADAM15 proteins;

[0111] (7) Inhibition of cell proliferation (e.g., tumor cells); and / or

[0112] (8) Inhibition of tumor growth.

[0113] In a specific embodiment, the antibody or its antigen-binding fragment according to any one of the embodiments may comprise an invariant region of human immunoglobulin, or an invariant region derived from human immunoglobulin.

[0114] In certain embodiments, the heavy chain of the antibody or its antigen-binding fragment comprises a heavy chain constant region of human immunoglobulin (e.g., IgG1, IgG2, IgG3, or IgG4) or a heavy chain constant region derived from human immunoglobulin. In certain embodiments, the heavy chain of the antibody or its antigen-binding fragment comprises a wild-type Fc region or a mutated or chemically modified Fc region having an altered effector function (e.g., reduced ADCC activity) compared to said wild-type Fc region. In certain exemplary embodiments, the antibody of the disclosure or its antigen-binding fragment comprises a variant of the human IgG1 heavy chain constant region, said variant having the following substitutions compared to the wild-type sequence from which said variant is derived: Leu234Ala, Leu235Ala, and Gly237Ala (positions according to the EU numbering system). In these embodiments, the antibody of the disclosure or its antigen-binding fragment has reduced ADCC activity. In a specific embodiment, the heavy chain of the antibody or its antigen-binding fragment comprises the sequence presented in SEQ ID NO: 5 or a variant thereof, said variant having a conservative substitution of up to 20 amino acids compared to SEQ ID NO: 5 (e.g., a conservative substitution of up to 15, up to 10, or up to 5 amino acids; e.g., a conservative substitution of 1, 2, 3, 4, or 5 amino acids). In a specific embodiment, the heavy chain constant region (CH) presented in SEQ ID NO: 5 or its variant lacks C-terminal lysine.

[0115] In certain embodiments, the light chain of the antibody or its antigen-binding fragment comprises a light chain constant region of human immunoglobulin, or a light chain constant region derived from human immunoglobulin (e.g., κ or λ). In certain embodiments, the light chain of the antibody or its antigen-binding fragment comprises the sequence presented in SEQ ID NO: 6 or a variant thereof, said variant having a conservative substitution of up to 20 amino acids compared to SEQ ID NO: 6 (e.g., a conservative substitution of up to 15, up to 10, or up to 5 amino acids; e.g., a conservative substitution of 1, 2, 3, 4, or 5 amino acids).

[0116] In certain embodiments, the antibody or its antigen-binding fragment of the present disclosure comprises:

[0117] (1) a heavy chain comprising the VH presented in SEQ ID NO: 54 and the heavy chain invariant region (CH) presented in SEQ ID NO: 5; and a light chain comprising the VL presented in SEQ ID NO: 55 and the light chain invariant region (CL) presented in SEQ ID NO: 6;

[0118] (2) a heavy chain comprising the VH presented in SEQ ID NO: 52 and the heavy chain invariant region (CH) presented in SEQ ID NO: 5; and a light chain comprising the VL presented in SEQ ID NO: 53 and the light chain invariant region (CL) presented in SEQ ID NO: 6;

[0119] (3) a heavy chain comprising the VH presented in SEQ ID NO: 3 and the heavy chain invariant region (CH) presented in SEQ ID NO: 5; and a light chain comprising the VL presented in SEQ ID NO: 4 and the light chain invariant region (CL) presented in SEQ ID NO: 6;

[0120] (4) a heavy chain comprising the VH presented in SEQ ID NO: 1 and the heavy chain invariant region (CH) presented in SEQ ID NO: 5; and a light chain comprising the VL presented in SEQ ID NO: 2 and the light chain invariant region (CL) presented in SEQ ID NO: 6;

[0121] (5) a heavy chain comprising the VH presented in SEQ ID NO: 9 and the heavy chain invariant region (CH) presented in SEQ ID NO: 5; and a light chain comprising the VL presented in SEQ ID NO: 10 and the light chain invariant region (CL) presented in SEQ ID NO: 6;

[0122] (6) a heavy chain comprising the VH presented in SEQ ID NO: 7 and the heavy chain invariant region (CH) presented in SEQ ID NO: 5; and a light chain comprising the VL presented in SEQ ID NO: 8 and the light chain invariant region (CL) presented in SEQ ID NO: 6;

[0123] (7) a heavy chain comprising the VH presented in SEQ ID NO: 50 and the heavy chain invariant region (CH) presented in SEQ ID NO: 5; and a light chain comprising the VL presented in SEQ ID NO: 51 and the light chain invariant region (CL) presented in SEQ ID NO: 6; or

[0124] (8) A heavy chain comprising the VH presented in SEQ ID NO: 48 and the heavy chain invariant region (CH) presented in SEQ ID NO: 5; and a light chain comprising the VL presented in SEQ ID NO: 49 and the light chain invariant region (CL) presented in SEQ ID NO: 6.

[0125] In certain embodiments, the antibody or its antigen-binding fragment of the present disclosure comprises:

[0126] (1) A heavy chain having the sequence presented in sequence number 88 and a light chain having the sequence presented in sequence number 89;

[0127] (2) A heavy chain having the sequence presented in SEQ ID NO: 40 and a light chain having the sequence presented in SEQ ID NO: 41; or,

[0128] (3) A heavy chain having the sequence presented in SEQ ID NO: 42 and a light chain having the sequence presented in SEQ ID NO: 43; or,

[0129] (4) A heavy chain having the sequence presented in sequence number 86 and a light chain having the sequence presented in sequence number 87.

[0130] In certain embodiments, a heavy chain N-terminal glutamine having the sequence or a variant thereof presented in SEQ ID NO: 88, 40, 42 or 86 undergoes a cyclization reaction to form pyroglutamic acid or pyroglutamate.

[0131] In certain embodiments, the antibody or its antigen-binding fragment according to any one of the embodiments is a murine antibody, a chimeric antibody, or a humanized antibody.

[0132] In a specific embodiment, the variable region of the antibody or its antigen-binding fragment according to any one of the embodiments is a variable region of human origin.

[0133] In a specific embodiment, the antibody or its antigen-binding fragment according to any one of the embodiments is selected from ScFv, Fab, Fab', F(ab')2, Fab'-SH, Fv fragment, dsFv (disulfide-linked Fv), diabody, bispecific antibody and multispecific antibody.

[0134] In certain embodiments, the antibody or its antigen-binding fragment according to any one of the embodiments has a label. In certain embodiments, the antibody or its antigen-binding fragment has a detectable label, for example, an enzyme (e.g., horseradish peroxidase), a radionuclide, a fluorescent dye, a luminescent substance (e.g., a chemiluminescent substance), or biotin.

[0135] The present disclosure also provides for the use of the antibody or antigen-binding fragment or the pharmaceutical composition thereof provided herein for the treatment of tumors.

[0136] Derived Antibodies (derived antibodies)

[0137] The antibody or its antigen-binding fragment of the present disclosure may be derivatized and, for example, linked to another molecule (e.g., another polypeptide or protein). Generally, the derivatization (e.g., labeling) of the antibody or its antigen-binding fragment does not have an adverse effect on its binding to ADAM9, specifically human ADAM9. Accordingly, the antibody or its antigen-binding fragment of the present disclosure is also intended to include such derivatized forms. For example, the antibody or its antigen-binding fragment of the present disclosure may be functionally linked (through chemical conjugation, genetic fusion, non-covalent bonding, or other means) to one or more other groups of molecules, for example, another antibody (e.g., bispecific antibody formation), detection reagents, pharmaceutical reagents, and / or proteins or polypeptides (e.g., avidin or polyhistidine tags) capable of mediating the binding of the antibody or antigen-binding fragment to another molecule.

[0138] As one of the antibody derivatives, the present disclosure provides a conjugate comprising the antibody of the present disclosure or its antigen-binding fragment and a conjugate moiety.

[0139] In certain embodiments, the junction moiety is selected from detectable labels. The detectable labels of the present disclosure may be any material that can be detected by fluorescence, spectroscopic, photochemical, biochemical, immunological, electrical, optical, or chemical means. These labels are well known in the art, examples of which are enzymes (e.g., horseradish peroxidase, alkaline phosphatase, β-galactosidase, urease, glucose oxidase, etc.), radionuclides (e.g., 3H, 125I, 35S, 14C, or 32P), fluorescent dyes (e.g., fluorescein isothiocyanate (FITC), fluorescein, tetramethylrhodamine isothiocyanate (TRITC), phycoerythrin (PE), Texas Red, rhodamine, quantum dots, or cyanine dye derivatives (e.g., Cy7 or Alexa 750)), acridinium ester compounds, magnetic beads (e.g., Dynabeads®), calorimetric labels such as colloidal gold or colored glass or plastic (e.g., polystyrene, polypropylene, latex, etc.) beads, and biotin for binding to avidin (e.g., streptavidin) modified with the above labels, but not limited thereto. In certain embodiments, such labels may be suitable for use in immunological detection (e.g., enzyme-linked immunoassay, radioimmunoassay, fluorescence immunoassay, chemiluminescence immunoassay, etc.). In certain embodiments, the detectable labels are selected from radioisotopes, fluorescent materials, luminescent materials, colored materials, or enzymes. In certain embodiments, the detectable labels described above may be linked to the antibodies of the present disclosure or their antigen-binding fragments via linkers of varying lengths to reduce potential steric hindrance.

[0140] In certain embodiments, the junction moiety is selected from therapeutic agents. In certain embodiments, the therapeutic agent is preferably an antitumor agent, e.g., a cytotoxic agent, a cytokine, a toxin, or a radionuclide.

[0141] In certain embodiments, the conjugated moiety is selected from materials that can improve the biological properties of the antibody (e.g., increase serum half-life). For example, the conjugated moiety may be a chemical group (e.g., polyethylene glycol (PEG), methyl or ethyl) or a glycosyl group.

[0142] As one of the above antibody derivatives, the present disclosure provides a multispecific antibody comprising the antibody of the present disclosure or its antigen-binding fragment.

[0143] In a specific embodiment, the multispecific antibody comprises the antibody of the present disclosure or its antigen-binding fragment as a first antigen-binding domain and further comprises at least one second antigen-binding domain for another target.

[0144] In a specific embodiment, the antigen-binding domain of the multispecific antibody retains its original antigen-binding specificity.

[0145] In a specific embodiment, the multispecific antibody is a bispecific antibody, a trispecific antibody, or a quadrupspecific antibody.

[0146] As one of the above antibody derivatives, the present disclosure provides a chimeric antigen receptor comprising the antibody of the present disclosure or its antigen-binding fragment. In certain embodiments, the chimeric antigen receptor comprises the antibody of the present disclosure or its antigen-binding fragment (e.g., ScFv) as an extracellular antigen-binding domain that specifically binds to ADAM9, and a transmembrane domain and one or more intracellular T cell signaling domains. The present disclosure also provides a host cell (e.g., immune cell, such as T lymphocyte, NK cell, DC cell, or macrophage) containing or expressing the chimeric antigen receptor.

[0147] Antibody production

[0148] The antibodies of the present disclosure may be prepared by various methods known in the art, for example, by genetic engineering recombination techniques. For example, DNA molecules encoding the heavy and light chain genes of the antibodies of the present disclosure are obtained by chemical synthesis or PCR amplification. The obtained DNA molecules are inserted into an expression vector and then transfected into host cells. Then, the transfected host cells are cultured under specific conditions and the antibodies of the present disclosure are expressed.

[0149] The antigen-binding fragment of the present disclosure can be obtained by hydrolyzing an intact antibody molecule (Morimoto et al. , J. Biochem . Biophys . Methods 24: 107-117 (1992) and Brennan et al. , Science See 229: 81 (1985)). In addition, these antigen-binding fragments can also be produced directly from recombinant host cells (Hudson, Curr . Opin . Immunol . 11: 548-557 (1999); Little et al. , Immunol Today (Reviewed in , 21: 364-370 (2000)). For example, the Fab' fragment can be obtained directly from a host cell; and the Fab' fragment can be chemically conjugated to form the F(ab')2 fragment (Carter et al. , Bio / Technology , 10: 163-167 (1992)). Furthermore, Fv, Fab, or F(ab')2 fragments can also be obtained directly through isolation from recombinant host cell cultures. Other techniques for preparing these antigen-binding fragments are well known to those skilled in the art.

[0150] Accordingly, in another aspect, the present disclosure provides an isolated nucleic acid molecule comprising a nucleotide sequence encoding the antibody of the present disclosure or its antigen-binding fragment, or its heavy chain variable region and / or light chain variable region. According to codon degeneracy in the art, in certain embodiments, said nucleotide sequence may be substituted according to codon degeneracy. In certain embodiments, said nucleotide sequence is codon-optimized.

[0151] In a specific embodiment, the isolated nucleic acid molecule comprises a nucleic acid molecule encoding a heavy chain variable region of an antibody and / or a nucleic acid molecule encoding a light chain variable region of an antibody, wherein the nucleic acid molecule encoding a heavy chain variable region of the antibody comprises (i) a nucleotide sequence presented in SEQ ID NO: 92, (ii) a sequence substantially identical to SEQ ID NO: 92 (e.g., a sequence having at least about 85%, 90%, 95%, 99% or more sequence identity with respect to SEQ ID NO: 92, or a sequence having one or more nucleotide substitutions), or (iii) a degenerate sequence of (i) or (ii); and / or a nucleic acid molecule encoding a light chain variable region of the antibody comprises (iv) a nucleotide sequence presented in SEQ ID NO: 93, (v) a sequence substantially identical to SEQ ID NO: 93 (e.g., a sequence having at least about 85%, 90%, 95%, 99% or more sequence identity with SEQ ID NO: 93, or a sequence having one or more nucleotide substitutions), or (vi) a degenerate sequence of (iv) or (v).

[0152] In a specific embodiment, the isolated nucleic acid molecule comprises a nucleic acid molecule encoding a heavy chain variable region of an antibody and / or a nucleic acid molecule encoding a light chain variable region of an antibody, wherein the nucleic acid molecule encoding a heavy chain variable region of the antibody comprises (i) a nucleotide sequence presented in SEQ ID NO: 44, (ii) a sequence substantially identical to SEQ ID NO: 44 (e.g., a sequence having at least about 85%, 90%, 95%, 99% or more sequence identity with respect to SEQ ID NO: 44, or a sequence having one or more nucleotide substitutions), or (iii) the degenerate sequence of (i) or (ii); and / or a nucleic acid molecule encoding a light chain variable region of the antibody comprises (iv) a nucleotide sequence presented in SEQ ID NO: 45, (v) a sequence substantially identical to SEQ ID NO: 45 (e.g., a sequence having at least about 85%, 90%, 95%, 99% or more sequence identity with SEQ ID NO: 45, or a sequence having one or more nucleotide substitutions), or (vi) a degenerate sequence of (iv) or (v).

[0153] In a specific embodiment, the isolated nucleic acid molecule comprises a nucleic acid molecule encoding a heavy chain variable region of an antibody and / or a nucleic acid molecule encoding a light chain variable region of an antibody, wherein the nucleic acid molecule encoding a heavy chain variable region of the antibody comprises (i) a nucleotide sequence presented in SEQ ID NO: 46, (ii) a sequence substantially identical to SEQ ID NO: 46 (e.g., a sequence having at least about 85%, 90%, 95%, 99% or more sequence identity with respect to SEQ ID NO: 46, or a sequence having one or more nucleotide substitutions), or (iii) the degenerate sequence of (i) or (ii); and / or a nucleic acid molecule encoding a light chain variable region of the antibody comprises (iv) a nucleotide sequence presented in SEQ ID NO: 47, (v) a sequence substantially identical to SEQ ID NO: 47 (e.g., a sequence having at least about 85%, 90%, 95%, 99% or more sequence identity with SEQ ID NO: 47, or a sequence having one or more nucleotide substitutions), or (vi) a degenerate sequence of (iv) or (v).

[0154] In a specific embodiment, the isolated nucleic acid molecule comprises a nucleic acid molecule encoding a heavy chain variable region of an antibody and / or a nucleic acid molecule encoding a light chain variable region of an antibody, and the nucleic acid molecule encoding a heavy chain variable region of the antibody comprises (i) a nucleotide sequence presented in SEQ ID NO: 90, (ii) a sequence substantially identical to SEQ ID NO: 90 (e.g., a sequence having at least about 85%, 90%, 95%, 99% or more sequence identity with respect to SEQ ID NO: 90, or a sequence having one or more nucleotide substitutions), or (iii) the degenerate sequence of (i) or (ii); and / or a nucleic acid molecule encoding a light chain variable region of the antibody comprises (iv) a nucleotide sequence presented in SEQ ID NO: 91, (v) a sequence substantially identical to SEQ ID NO: 91 (e.g., a sequence having at least about 85%, 90%, 95%, 99% or more sequence identity with SEQ ID NO: 91, or a sequence having one or more nucleotide substitutions), or (vi) a degenerate sequence of (iv) or (v).

[0155] In another aspect of the present disclosure, the present disclosure provides a vector (e.g., a cloning vector or an expression vector) comprising a nucleic acid molecule isolated from the present disclosure. In certain embodiments, the vector of the present disclosure is, for example, a plasmid, a cosmid, a phage, a lentivirus, etc. In certain embodiments, the vector may express the antibody of the present disclosure or its antigen-binding fragment in a subject (e.g., a mammal, e.g., a human).

[0156] In a specific embodiment, the vector comprises a first nucleotide sequence encoding a heavy chain or a heavy chain variable region of the antibody of the present disclosure or its antigen-binding fragment, and a second nucleotide sequence encoding a light chain or its light chain variable region, wherein the first nucleotide sequence and the second nucleotide sequence are in the same vector or different vectors. Where the first nucleotide sequence and the second nucleotide sequence are in different vectors, the vector of the present disclosure comprises a first vector comprising the first nucleotide sequence and a second vector comprising the second nucleotide sequence.

[0157] In certain embodiments, the antibody of the present disclosure or its antigen-binding fragment may be used to construct a chimeric antigen receptor (CAR), which comprises an extracellular antigen-binding domain (e.g., ScFv) that specifically binds to ADAM9, a transmembrane domain, and one or more intracellular T cell signaling domains. In such embodiments, the isolated nucleic acid molecule of the present disclosure may comprise a nucleotide sequence encoding a chimeric antigen receptor, said nucleotide sequence encoding a chimeric antigen receptor further comprises a nucleotide sequence encoding an antibody of the present disclosure or its antigen-binding fragment (e.g., ScFv). In certain embodiments, the isolated nucleic acid molecule of the present disclosure encodes a chimeric antigen receptor comprising an antigen-binding fragment (e.g., ScFv) of an antibody of the present disclosure.

[0158] In certain embodiments, the antibody of the present disclosure or its antigen-binding fragment may be used to produce a chimeric antigen receptor-modified immune cell, which comprises a chimeric antigen receptor (CAR) and an immune cell (e.g., T lymphocyte, NK cell, DC cell, or macrophage).

[0159] In another aspect of the present disclosure, the present disclosure provides a host cell comprising the isolated nucleic acid molecule of the present disclosure or the vector of the present disclosure. The host cell is a eukaryotic cell (e.g., mammalian cell, insect cell, or yeast cell) or a prokaryotic cell (e.g., E. coli ( Escherichia coli Suitable eukaryotic cells include, but are not limited to, NS0 cells, Vero cells, Hela cells, COS cells, CHO cells, ExpiCHO cells, HEK293 cells, Expi293 cells, BHK cells, and MDCKII cells. Suitable insect cells include, but are not limited to, Sf9 cells. In certain embodiments, the host cell of the present disclosure is a mammalian cell, e.g., CHO (e.g., CHO-K1, CHO-S, CHO DXB11, ExpiCHO, CHO DG44, or CHO-EBNA).

[0160] In certain embodiments, the host cell of the present disclosure may be a chimeric antigen receptor T cell (CAR-T). In such embodiments, the isolated nucleic acid molecule contained in the host cell may comprise a nucleotide sequence encoding a chimeric antigen receptor, and the nucleotide sequence encoding the chimeric antigen receptor further comprises a nucleotide sequence encoding the antibody of the present disclosure or its antigen-binding fragment (e.g., ScFv). In certain embodiments, the isolated nucleic acid molecule contained in the host cell codes for a chimeric antigen receptor comprising an antigen-binding fragment (e.g., ScFv) of the antibody of the present disclosure.

[0161] In another aspect of the present disclosure, a method for producing an antibody of the present disclosure or its antigen-binding fragment is provided, the method comprising the steps of culturing a host cell of the present disclosure under conditions allowing the expression of said antibody or its antigen-binding fragment, and recovering said antibody or its antigen-binding fragment from a culture of said cultured host cell.

[0162] therapeutic use

[0163] In another aspect of the present disclosure, a pharmaceutical composition is provided comprising the antibody of the present disclosure or its antigen-binding fragment, a nucleic acid molecule, a vector, a host cell, a conjugate, a multispecific antibody, or a chimeric antigen receptor or a host cell expressing said chimeric antigen receptor, and a pharmaceutically acceptable carrier and / or excipient.

[0164] In certain embodiments, the pharmaceutical composition of the present disclosure comprises the antibody of the present disclosure or its antigen-binding fragment, and a pharmaceutically acceptable carrier and / or excipient.

[0165] In certain embodiments, the pharmaceutical composition of the present disclosure comprises a vector or host cell of the present disclosure and a pharmaceutically acceptable carrier and / or excipient. In such embodiments, the isolated nucleic acid molecule contained in the vector comprises a nucleotide sequence encoding a chimeric antigen receptor, and said nucleotide sequence encoding the chimeric antigen receptor further comprises a nucleotide sequence encoding the antibody of the present disclosure or its antigen-binding fragment (e.g., ScFv); and said host cell comprises the isolated nucleic acid molecule or vector described above. In certain embodiments, said isolated nucleic acid molecule codes for a chimeric antigen receptor comprising an antigen-binding fragment (e.g., ScFv) of the antibody of the present disclosure. In certain embodiments, said host cell is an immune cell, e.g., a T cell. In certain embodiments, said host cell is a chimeric antigen receptor T cell (CAR-T).

[0166] In certain embodiments, the pharmaceutical composition may also include an additional pharmaceutical active agent. In certain embodiments, the additional pharmaceutical active agent is a drug having anti-tumor activity. In certain embodiments, the additional pharmaceutical active agent is selected from ADAM9 inhibitors, EGFR inhibitors, HER2 inhibitors, HER3 inhibitors, HER4 inhibitors, IGFR-1 inhibitors, mTOR inhibitors, PI3 kinase inhibitors, c-met inhibitors, or VEGF inhibitors, chemotherapy agents, or any combination thereof. In certain embodiments, the antibody of the present disclosure or its antigen-binding fragment and the additional pharmaceutical active agent are provided as separate components or as mixed components. Accordingly, the antibody of the present disclosure or its antigen-binding fragment and the additional pharmaceutical active agent may be administered simultaneously, individually, or sequentially.

[0167] In certain embodiments, an antibody or its antigen-binding fragment, a nucleic acid molecule, a vector, a host cell, a conjugate, a multispecific antibody, or a chimeric antigen receptor or a host cell expressing said chimeric antigen receptor among the pharmaceutical compositions of the present disclosure is sufficient to perform the following (e.g., in a subject):

[0168] (a) Inhibition of cell proliferation (e.g., tumor cells);

[0169] (b) inhibition of tumor growth;

[0170] (c) Induction and / or increase of antibody-dependent cytotoxic activity;

[0171] (d) Inhibition of ADAM9-mediated signaling;

[0172] (e) Prevention and / or treatment of ADAM9-mediated diseases / disorders; or

[0173] (f) Any combination of (a) to (e) above.

[0174] In certain embodiments, the ADAM9-mediated disease / disorder is a tumor, e.g., an ADAM9-expressing tumor. In certain embodiments, the tumor is selected from glioma, cervical cancer, oral squamous cell carcinoma, liver cancer, melanoma, prostate cancer, pancreatic ductal adenocarcinoma, gastric cancer, lung cancer such as non-small cell lung cancer, pancreatic cancer, ovarian cancer, colorectal cancer, head and neck cancer, breast cancer such as triple-negative breast cancer, or any combination thereof.

[0175] In another aspect of the present disclosure, use of the antibody or its antigen-binding fragment of the present disclosure, nucleic acid molecule, vector, host cell, conjugate, multispecific antibody, chimeric antigen receptor or host cell expressing said chimeric antigen receptor, or pharmaceutical composition is provided in the preparation of a drug for the inhibition of cell proliferation, or for the prevention and / or treatment and / or adjuvant treatment of a tumor, neurodegenerative disease, retinal disease or inflammation.

[0176] In certain embodiments, the drug is used to inhibit the proliferation of ADAM9-expressing cells (e.g., tumor cells).

[0177] In another aspect of the present disclosure, a method for inhibiting cell proliferation is provided, said method comprising the step of contacting said cell with an antibody of the present disclosure or its antigen-binding fragment, a nucleic acid molecule, a vector, a host cell, a conjugate, a multispecific antibody, a chimeric antigen receptor or a host cell expressing said chimeric antigen receptor, or a pharmaceutical composition. In a specific embodiment, said cell is an ADAM9-expressing cell, e.g., a tumor cell.

[0178] In another aspect of the present disclosure, a method for preventing and / or treating and / or adjuvantly treating a tumor, neurodegenerative disease, retinal disease, or inflammation in a subject is provided, the method comprises the step of administering an effective amount of the antibody or its antigen-binding fragment of the present disclosure, a nucleic acid molecule, a vector, a host cell, a conjugate, a multispecific antibody, a chimeric antigen receptor or a host cell expressing said chimeric antigen receptor, or a pharmaceutical composition to a subject in need thereof.

[0179] In a specific embodiment, the method further comprises the step of applying a second therapy selected from surgery, chemotherapy, radiation therapy, immunotherapy, gene therapy, DNA therapy, RNA therapy, nanotherapy, viral therapy, adjuvant therapy, and any combination thereof to the subject. In a specific embodiment, the second therapy may be applied simultaneously with, individually with, or sequentially with the method.

[0180] In any one of the above embodiments, the antibody of the present disclosure or its antigen-binding fragment, nucleic acid molecule, vector, host cell, conjugate, multispecific antibody, chimeric antigen receptor or host cell expressing said chimeric antigen receptor, or tumor associated with the pharmaceutical composition may have any tumor type. In a specific embodiment, the antibody of the present disclosure or its antigen-binding fragment, nucleic acid molecule, vector, host cell, conjugate, multispecific antibody, chimeric antigen receptor or host cell expressing said chimeric antigen receptor, or tumor associated with the pharmaceutical composition is an ADAM9-positive tumor. In certain embodiments, the antibody of the present disclosure or its antigen-binding fragment, nucleic acid molecule, vector, host cell, conjugate, multispecific antibody, chimeric antigen receptor or host cell expressing said chimeric antigen receptor, or tumor related to the pharmaceutical composition is selected from glioma, cervical cancer, oral squamous cell carcinoma, liver cancer, melanoma, prostate cancer, pancreatic ductal adenocarcinoma, gastric cancer, lung cancer such as non-small cell lung cancer, pancreatic cancer, ovarian cancer, colorectal cancer, head and neck cancer, breast cancer such as triple-negative breast cancer, or any combination thereof.

[0181] The antibody or its antigen-binding fragment of the present disclosure, or the pharmaceutical composition of the present disclosure, may be formulated into any formulation known in the pharmaceutical art, e.g., tablets, pills, suspensions, emulsions, solutions, gels, capsules, powders, granules, elixirs, troches, suppositories, injectables (including injectable solutions, sterile powders for injection, and concentrated solutions for injection), inhalants, sprays, etc. Preferred formulations depend on the intended mode of administration and therapeutic use. The pharmaceutical composition of the present disclosure must be sterile and stable under manufacturing and storage conditions. One preferred formulation is an injectable. Such an injectable may be a sterile injectable solution. For example, a sterile injectable solution may be prepared by the method described below: a required dose of the antibody of the present disclosure is incorporated into a suitable solvent, optionally other desired components (including but not limited to pH adjusters, surfactants, adjuvants, ionic strength enhancers, isotonic agents, preservatives, diluents, or any combination thereof) are added simultaneously, and then sterilized by filtration. Additionally, the sterile injectable solution may be prepared as a sterile lyophilized powder (e.g., by vacuum drying or freeze-drying) for ease of storage and use. Such sterile lyophilized powder may be dispersed in a suitable carrier, e.g., sterile pyrogen-free water, before use.

[0182] Additionally, the antibody or its antigen-binding fragment of the present disclosure may be present in a pharmaceutical composition in a unit dosage form for ease of administration.

[0183] The antibody or its antigen-binding fragment or pharmaceutical composition of the present disclosure may be administered by any suitable method known in the art, including but not limited to oral, buccal, sublingual, ocular, topical, parenteral, rectal, intrathecal, intravesical, inguinal, intravesical, topical (e.g., powder, ointment, or drops), or nasal routes. However, for many therapeutic uses, the preferred route / mode of administration is parenteral administration (e.g., intravenous injection, subcutaneous injection, intraperitoneal injection, or intramuscular injection). Those skilled in the art should understand that the route and / or mode of administration will vary depending on the intended purpose. In one preferred embodiment, the antibody or its antigen-binding fragment or pharmaceutical composition of the present disclosure is administered by intravenous infusion or injection.

[0184] The pharmaceutical composition of the present disclosure may comprise a "therapeutically effective amount" or a "prophylactically effective amount" of the antibody or its antigen-binding fragment of the present disclosure. A "prophylactically effective amount" means an amount sufficient to prevent, stop, or delay the onset of disease. A "therapeutically effective amount" means an amount sufficient to treat or at least partially prevent disease and its complications in a patient suffering from disease. The therapeutically effective amount of the antibody or its antigen-binding fragment of the present disclosure may vary depending on the following factors: the severity of the disease to be treated, the overall condition of the patient's own immune system, the patient's general circumstances, such as age, weight, and sex, the mode of drug administration, other concomitant treatments, etc.

[0185] In the present disclosure, the administration regimen may be adjusted to obtain an optimal desired response (e.g., a therapeutic or prophylactic response). For example, a single dose may be administered, multiple doses may be administered over a period of time, or the dose may be proportionally reduced or increased depending on the level of urgency of the treatment situation.

[0186] In the present disclosure, the subject may be a mammal, for example, a human.

[0187] Detection applications

[0188] The antibody or its antigen-binding fragment of the present disclosure can specifically bind to ADAM9 and thus can be used to detect the presence or level of ADAM9 in a sample.

[0189] Accordingly, in another aspect, the present disclosure provides a kit comprising the antibody of the present disclosure or its antigen-binding fragment. In a specific embodiment, the antibody of the present disclosure or its antigen-binding fragment has a detectable label. In a preferred embodiment, the kit further comprises a second antibody that specifically recognizes the antibody of the present disclosure or its antigen-binding fragment. Preferably, the second antibody further comprises a detectable label.

[0190] In the present disclosure, the detectable label may be any substance detectable by fluorescence, spectroscopic, photochemical, biochemical, immunological, electrical, optical, or chemical means. It is particularly preferred that such a label be suitable for use in immunological detection (e.g., enzyme-linked immunoassay, radioimmunoassay, fluorescence immunoassay, chemiluminescence immunoassay, etc.). These labels are well known in the art and include enzymes (e.g., horseradish peroxidase, alkaline phosphatase, β-galactosidase, urease, glucose oxidase, etc.), radionuclides (e.g., 3H, 125I, 35S, 14C, or 32P), fluorescent dyes (e.g., fluorescein isothiocyanate (FITC), fluorescein, tetramethylrhodamine isothiocyanate (TRITC), phycoerythrin (PE), Texas Red, rhodamine, quantum dots, or cyanine dye derivatives (e.g., Cy7 or Alexa 750)), acridinium ester compounds, magnetic beads (e.g., Dynabeads®), calorimetric labels such as colloidal gold or colored glass or plastic (e.g., polystyrene, polypropylene, latex, etc.) beads, and biotin for binding to avidin (e.g., streptavidin) modified with said labels Includes, but is not limited to. In certain embodiments, the detectable label described above may be linked to the antibody of the present disclosure through linkers of various lengths to reduce potential steric hindrance.

[0191] In another aspect, the present disclosure provides a method for detecting the presence or level of ADAM9 in a sample, said method comprising the step of using the antibody of the present disclosure or its antigen-binding fragment. In a preferred embodiment, the antibody of the present disclosure or its antigen-binding fragment further comprises a detectable label. In another preferred embodiment, said method further comprises the step of detecting the antibody of the present disclosure or its antigen-binding fragment using a reagent having a detectable label. said method may be used for diagnostic or non-diagnostic purposes (e.g., said sample is a cell sample rather than a sample derived from a patient).

[0192] In a specific embodiment, the method comprises the steps of contacting a sample with said antibody or said antigen-binding fragment under conditions that allow the formation of a complex between said antibody or said antigen-binding fragment and ADAM9, and detecting the formation of said complex.

[0193] Considering that ADAM9 is not expressed or is expressed at low levels in normal tissues, and is expressed or is expressed at high levels in some cancers, a tumor can be diagnosed by detecting the presence or level of ADAM9 in a sample. Accordingly, in certain embodiments, the method is used to diagnose a tumor, e.g., an ADAM9-positive tumor, e.g., glioma, cervical cancer, oral squamous cell carcinoma, liver cancer, melanoma, prostate cancer, pancreatic ductal adenocarcinoma, gastric cancer, lung cancer, e.g., non-small cell lung cancer, pancreatic cancer, ovarian cancer, colorectal cancer, head and neck cancer, breast cancer, e.g., triple-negative breast cancer, or any combination thereof.

[0194] In a specific embodiment, the method comprises the steps of detecting the expression level of ADAM9 in a test sample derived from a subject and comparing the expression level with a reference value (e.g., a healthy control group), wherein an increase in the expression level compared to the reference value indicates a tumor.

[0195] In another aspect, the use of the antibody or its antigen-binding fragment of the present disclosure is provided in the manufacture of a kit for the detection of the presence or level of ADAM9 in a sample and / or the diagnosis of a tumor.

[0196] In another aspect, the present disclosure provides a diagnostic or therapeutic kit comprising the antibody of the present disclosure or its antigen-binding fragment, a nucleic acid molecule, a vector, a host cell, a conjugate or a multispecific antibody, and instructions for use. The kit may also include a delivery device for local administration. The delivery device includes a drug-loaded syringe or a needle-free device. Effects of the invention

[0197] The antibody of the present disclosure exhibits high binding affinity and very strong specificity for ADAM9 and does not possess ADCC activity, thereby effectively avoiding side effects associated with ADCC function. The antibody of the present disclosure can inhibit tumor cell proliferation, and thus has potential for the prevention and / or treatment of tumors. Furthermore, the antibody of the present disclosure is a humanized antibody that does not induce an immunogenic response and can be safely administered to human subjects. Therefore, the antibody of the present disclosure has significant clinical value. Brief explanation of the drawing

[0198] Fig. 1: Results of affinity detection of anti-human ADAM9 murine antibody against EBC-1 cells. Fig. 2: Results of affinity detection of anti-human ADAM9 chimeric antibody and humanized antibody against EBC-1 cells. Fig. 3: Results of affinity detection of anti-human ADAM9 chimeric antibody and humanized antibody against HCC1806 cells. Fig. 4: Results of detection of endocytic activity of anti-human ADAM9 chimeric antibody and humanized antibody. Fig. 5: Results of specificity detection of anti-human ADAM9 humanized antibody. Specific details for implementing the invention

[0199] abbreviation

[0200]

[0201] definition

[0202] In this disclosure, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art unless otherwise specified. Furthermore, laboratory operation steps such as cell culture, biochemistry, nucleic acid chemistry, and immunology used herein are all common steps widely used in the field. Moreover, to aid in understanding this disclosure, definitions and explanations of related terms are provided below.

[0203] As used herein, the term “antibody” is used in its broadest sense and includes, but is not limited to, various antibody structures including monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, insofar as they exhibit the desired antigen-binding activity. For example, the “antibody” may be an immunoglobulin molecule composed of two pairs of polypeptide chains, each pair having a light chain (LC) and a heavy chain (HC). The antibody light chains may be classified as κ (kappa) light chains and λ (lambda) light chains. The heavy chains may be classified as μ, δ, γ, α, or ε, and the isotypes of the antibody are defined as IgM, IgD, IgG, IgA, and IgE, respectively. Within the light and heavy chains, the variable and constant regions are connected by a "J" region composed of about 12 or more amino acids, wherein the heavy chain further includes a "D" region composed of about 3 or more amino acids. Each heavy chain consists of a heavy chain variable region (VH) and a heavy chain constant region (CH). The heavy chain constant region consists of three domains (CH1, CH2, and CH3). Each light chain consists of a light chain variable region (VL) and a light chain constant region (CL). The light chain constant region consists of a single domain, CL. The constant domain does not directly participate in antibody-antigen binding but exhibits various effector functions. For example, the constant domain can mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system. The VH and VL regions can also be subdivided into regions of high variability (called Complementarity Decision 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 order FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4 from the amino terminus to the carboxy terminus. The variable regions (VH and VL) of each heavy / light chain pair form antigen-binding sites. The distribution of amino acids in various regions or domains is described in Kabat, *Sequences of Proteins of Immunological Interest* (National Institutes of Health, Bethesda, Md. (1987 and 1991)), or Chothia & Lesk (1987). J. Mol . Biol . 196: 901-917; and Chothia et al. (1989) Nature 342: Follows the definition of 878-883.

[0204] Unless clearly otherwise indicated in the context, where the term “antibody” is used herein, it includes not only the intact antibody but also antigen-binding fragments of the antibody.

[0205] The term “antibody” also includes embodiments in which the constant region of the heavy chain contains C-terminal lysine, or lacks C-terminal lysine or C-terminal glycine-lysine dipeptide. The term also includes embodiments in which the N-terminal amino acid of the variable region of the antibody is cyclized with pyroglutamate. Accordingly, in a composition comprising the antibody disclosed herein, the antibody may independently contain C-terminal lysine, may lack C-terminal lysine, may lack C-terminal glycine-lysine, and / or may contain an N-terminal amino acid cyclized with N-terminal glutamine or glutamic acid or pyroglutamic acid.

[0206] As used herein, the terms "complementarity determining region" or "CDR" refer to amino acid residues within the variable region of an antibody responsible for antigen binding. The precise boundaries of these amino acid residues are determined according to various numbering systems known in the art, for example, the AbM numbering system (Martin ACR, Cheetham JC, Rees AR (1989) Modelling antibody hypervariable loops: A combined algorithm. Proc Natl Acad Sci USA 86: 9268-9272) or IMGT numbering system (Lefranc et al. , Dev. Comparat . Immunol . It can be defined according to 27: 55-77, 2003). For a given antibody, a person skilled in the art will be able to easily identify the CDRs defined by the numbering system. In addition, the correspondence between different numbering systems is well known to a person skilled in the art (e.g., Lefranc et al. , Dev . Comparat . Immunol . See 27: 55-77, 2003).

[0207] In the present disclosure, the CDRs included in the antibody or its antigen-binding fragment of the present disclosure may be determined according to various numbering systems known in the art. In certain embodiments, the CDRs included in the antibody or its antigen-binding fragment of the present disclosure are preferably determined by the IMGT, Kabat, Chothia, or AbM numbering system.

[0208] The following general rules (published on www.bioinf.org.uk by Professor Andrew CR Martin’s research group) can be used to define CDRs in antibody sequences, which include amino acids that specifically interact with the amino acids constituting the antigenic epitope bound by the antibody. Although rarely these general invariant features appear; Cys residues are the most conserved features.

[0209]

[0210] V H The entire amino acid sequence of is generally numbered according to Kabat, and the three CDRs within the variable region may be defined according to any one of the numbering systems. In a specific embodiment, V H In this sequence, amino acid positions may be numbered sequentially from amino acid position 1 to the end of the sequence, or numbered according to Kabat. Unless otherwise specified, V as described herein H and V L The amino acid positions are defined according to sequential numbering.

[0211] Amino acid positions in the heavy chain constant region may be numbered sequentially from amino acid position 1 to the end of the sequence, or numbered according to the Eu numbering system. The amino acid sequence of the IgG1 heavy chain constant region comprises 330 amino acids numbered sequentially from number 1 to 330. The corresponding sequence numbered according to the Eu numbering system begins at position 118 and ends at position 447. Unless otherwise specified, the amino acid positions of the heavy and light chains described herein are defined according to sequential numbering.

[0212] As used herein, the terms "framework region" or "FR" residue refer to amino acid residues within the antibody variable region other than the CDR residues defined above.

[0213] The term "antibody" is not limited to any specific method of producing antibodies. For example, the antibodies include recombinant antibodies, monoclonal antibodies, and polyclonal antibodies. The antibodies may be various isotypes of antibodies, for example, IgG (e.g., IgG1, IgG2, IgG3, or IgG4 subtypes), IgA1, IgA2, IgD, IgE, or IgM antibodies.

[0214] As used herein, the term “antigen-binding fragment” of an antibody means a molecule other than the intact antibody that comprises a portion of the intact antibody and binds to the antigen to which the intact antibody binds, for example, the antigen-binding fragment may be a polypeptide of a fragment of a full-length antibody that retains the ability to specifically bind to the same antigen to which the full-length antibody binds, and / or competes with the full-length antibody for specific binding to the antigen, and is also referred to as an “antigen-binding moiety.” Generally, refer to Fundamental Immunology, Ch. 7, Paul, W., ed., 2nd ed., Raven Press, NY (1989), the entirety of which is incorporated herein by reference for all purposes. Antigen-binding fragments of antibodies may be produced by recombinant DNA technology or by enzymatic or chemical cleavage of an intact antibody. Non-limiting examples of antigen-binding fragments include Fab, Fab', Fab'-SH, F(ab')2, Fd, Fv, dAb, and complementarity determining region (CDR) fragments, short-chain antibodies (e.g., scFv), chimeric antibodies, diabodies, linear antibodies, nanobodies (Domantis technology), domain antibodies (Ablynx technology), and polypeptides comprising at least a portion of an antibody sufficient to confer specific antigen-binding ability to the polypeptide. Engineered antibody variants are Holliger et al. , 2005; Nat Biotechnol , 23: Reviewed in 1126-1136.

[0215] As used herein, the term "full-length antibody" refers to an antibody composed of two "full-length heavy chains" or "heavy chains" and two "full-length light chains" or "light chains." The "full-length heavy chains" or "heavy chains" refer to polypeptide chains composed of a heavy chain variable region (VH), a heavy chain constant region CH1 domain, a hinge region (HR), a heavy chain constant region CH2 domain, and a heavy chain constant region CH3 domain, in the direction from the N-terminus to the C-terminus. Furthermore, if the full-length antibody has an IgE isotype, the heavy chains optionally additionally comprise a heavy chain constant region CH4 domain. Preferably, the "full-length heavy chains" are polypeptide chains composed of VH, CH1, HR, CH2, and CH3 in the direction from the N-terminus to the C-terminus. "Whole-length light chain" or "light chain" is a polypeptide chain composed of a light chain variable region (VL) and a light chain constant region (CL) in the direction from the N-terminus to the C-terminus. Two pairs of whole-length antibody chains are linked together through disulfide bonds between CL and CH1, and disulfide bonds between the HRs of two whole-length heavy chains. The whole-length antibody of the present disclosure may be derived from a single species, e.g., human. It may also be a chimeric antibody or a humanized antibody. The whole-length antibody of the present disclosure comprises two antigen-binding sites formed by VH and VL pairs, each of which specifically recognizes / binds to the same antigen.

[0216] As used herein, the term “Fab fragment” means an antibody fragment composed of VL, VH, CL, and CH1 domains; the term “F(ab’)2 fragment” means an antibody fragment comprising two Fab fragments connected by a disulfide bond at a hinge region; the term “Fab’ fragment” means a fragment obtained by reducing the disulfide bond connecting two heavy chain fragments in the F(ab’)2 fragment, and comprising a whole light chain and heavy chain Fd fragment (composed of VH and CH1 domains); and the term “Fab’-SH” means a Fab fragment containing free sulfhydryl groups.

[0217] As used herein, the term “Fv fragment” refers to an antibody fragment consisting of the VL and VH domains of a single arm of an antibody. The Fv fragment is generally considered to be the minimum antibody fragment capable of forming a complete antigen-binding site. Generally, six CDRs are considered to confer antigen-binding specificity to the antibody. However, even if the affinity is lower than that of the complete binding site, a variable region (e.g., an Fd fragment containing only three CDRs specific to the antigen) can recognize and bind to the antigen.

[0218] As used herein, the term "Fc fragment" refers to an antibody fragment formed by connecting the second and third constant regions of the first heavy chain of an antibody and the second and third constant regions of the second heavy chain via a disulfide bond. The Fc fragment of an antibody has various functions but does not participate in antigen binding.

[0219] As used herein, the term "scFv" refers to a single polypeptide chain comprising VL and VH domains, said VL and VH are connected through a linker (e.g., Bird et al. , Science 242: 423-426 (1988); Huston et al. , Proc . Natl. Acad . Sci USA 85: 5879-5883 (1988); and see Pluckthun, The Pharmacology of Monoclonal Antibodies, vol. 113, Roseburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994). These scFv molecules may have a general structure: NH2-VL-linker-VH-COOH or NH2-VH-linker-VL-COOH. In the prior art, suitable linkers consist of a repeated GGGGS amino acid sequence or a variant thereof. For example, a linker having the amino acid sequence (GGGGS)4 can be used, and a variant thereof can also be used (Holliger et al. (1993), Proc . Natl . Acad . Sci USA 90: 6444-6448). Other linkers that can be used in this disclosure are 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 It is described in . In some cases, a disulfide bond may also exist between the VH and VL of scFv.

[0220] As used herein, the term "diabody" means that its VH and VL domains are expressed on a single polypeptide chain, but because the linker used is too short to form a pair between the two domains on the same chain, the domains are paired with a complementary domain on another chain to create two antigen-binding sites (e.g., Holliger P. et al. , Proc . Natl . Acad . Sci USA 90: 6444-6448 (1993), and Poljak RJ et al. , Structure 2: See 1121-1123 (1994).

[0221] All of the above antibody fragments retain the ability to specifically bind to the same antigen to which the full-length antibody binds, and / or compete with the full-length antibody for specific binding to the antigen. In this invention, a person skilled in the art may obtain an antigen-binding fragment of an antibody (e.g., the antibody fragment described above) from a given antibody (e.g., the antibody provided herein) using known conventional techniques and specifically screen the antigen-binding fragment of the antibody in the same manner as for the intact antibody.

[0222] As used herein, the term “multispecific antibody” means an antibody having various different antigen-binding specificities, including, for example, bispecific antibodies, trispecific antibodies, and quadrspecific antibodies. “Bispecific antibody” means an antibody having two different antigen-binding specificities, which is a conjugate or antibody analog formed by a first antibody (or a fragment thereof) and a second antibody (or a fragment thereof) through a conjugation arm; the conjugation method includes, but is not limited to, chemical reactions, gene fusion, and enzymatic reactions. “Multispecific antibody” includes, for example, trispecific antibodies and quadrspecific antibodies, wherein the trispecific antibody is an antibody having three distinct antigen-binding specificities and the quadrspecific antibody is an antibody having four distinct antigen-binding specificities.

[0223] As used herein, the terms "monoclonal antibody," "monoclonal Ab," and "mAb" have the same meaning and are used interchangeably. They refer to a group of highly homologous antibody molecules, that is, antibodies or fragments of antibodies derived from a group of identical antibody molecules, excluding possible naturally occurring spontaneous mutations. Such monoclonal antibodies have a high degree of specificity for a single epitope of an antigen. In contrast to monoclonal antibodies, polyclonal antibodies generally comprise at least two different antibodies, and these different antibodies generally recognize different epitopes of the antigen. Furthermore, the modifier "monoclonal" merely indicates that the antibody is characterized by being derived from a highly homologous group of antibodies and is not understood to require any specific method for the production of the antibody.

[0224] As used herein, the term “chimeric antibody” means an antibody in which a portion of the light chain or / and heavy chain is derived from one antibody (which may be derived from a specific species or belong to a specific antibody class or subclass), and another portion of the light chain or / and heavy chain is derived from another antibody (which may be derived from the same or different species or belong to the same or different antibody class or subclass), but nevertheless retains binding activity to a target antigen. For example, the term “chimeric antibody” may include an antibody in which the variable regions of the heavy and light chains are derived from a first antibody (e.g., an antibody of human origin), and the constant regions of the heavy and light chains are derived from a second antibody (e.g., an antibody of murine origin).

[0225] As used herein, the term “humanized antibody” means an antibody that can be prepared by replacing a portion of a human antibody with a portion of a non-human antibody prepared by immunizing a non-human mammal. Typically, all or part of the CDR region of the humanized antibody is derived from a non-human antibody (donor antibody), and all or part of the non-CDR region (e.g., variable region FR and / or constant region) is derived from a human immunoglobulin (receptor antibody). For example, a humanized antibody can be prepared by transplanting a CDR sequence derived from germlines of another mammalian species into a human framework sequence.

[0226] As used herein, the term “variant” means a polypeptide or peptide comprising an amino acid sequence altered by the substitution, deletion, or additional introduction of amino acid residues in relation to a polypeptide (including polypeptides). In some cases, the term “variant” also means a polypeptide or peptide modified (i.e., by covalent bonding any type of molecule to the polypeptide or peptide). For example, polypeptides may be modified by, but are not limited to, glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting / blocking groups, proteolytic cleavage, attachment to cell ligands or other proteins, etc. Derivatized polypeptides or peptides may be produced by chemical modification using techniques known to those skilled in the art, including but not limited to specific chemical cleavage, acetylation, formylation, and metabolic synthesis using tunicamycin. In addition, the variant has similar, identical, or improved functions compared to the polypeptide or peptide from which it is derived.

[0227] As used herein, the term "specific binding" refers to a non-random binding reaction between two molecules, such as the reaction between an antibody and its corresponding antigen. The strength or affinity of a specific binding interaction is the equilibrium dissociation constant (KD) or half-maximal effect concentration (EC₀) of the interaction. 50 It can be displayed as ).

[0228] The specific binding characteristics between two molecules can be determined using methods well known in the art. One method involves measuring the formation and dissociation rates of antigen-binding sites / antigen complexes. The "association rate constant" (ka or kon) and the "dissociation rate constant" (kdis or koff) can be calculated based on concentration and actual binding and dissociation rates (Malmqvist M, Nature See , 1993, 361: 186-187). The ratio of kdis / kon is equal to the dissociation constant KD (Davies et al. , Annual Rev Biochem (See , 1990; 59: 439-473). Any effective method can be used to measure the values ​​of KD, kon, and kdis. In certain embodiments, the dissociation constant can be measured using a bioluminescent interferometer (e.g., ForteBio Octet method). Additionally, the dissociation constant can be measured using surface plasmon resonance techniques (e.g., Biacore) or Kinexa.

[0229] As used herein, the term "vector" refers to a nucleic acid delivery vehicle into which a polynucleotide can be inserted. When the vector enables the expression of a protein encoded by the inserted polynucleotide, said vector is referred to as an expression vector. The vector may be introduced into a host cell by transformation, transduction, or transfection, and the genetic material elements carried by said vector may be expressed in said host cell. Vectors are well known to those skilled in the art and include, but are not limited to, plasmids; phages; cosmids; artificial chromosomes such as yeast artificial chromosomes (YACs), bacterial artificial chromosomes (BACs), or P1-derived artificial chromosomes (PACs); and phages such as λ phage or M13 phage and animal viruses. Animal viruses that may be used as vectors include, but are not limited to, retroviruses (including lentiviruses), adenoviruses, adeno-associated viruses, herpes viruses (e.g., herpes simplex virus), poxviruses, baculoviruses, papillomaviruses, and papovaviruses (e.g., SV40). The vector may include various elements controlling expression, including, but not limited to, a promoter sequence, a transcription initiation sequence, an enhancer sequence, a selection element, and a reporter gene. Additionally, the vector may further include a replication origin.

[0230] Expression and cloning vectors comprise a nucleic acid sequence that enables the vector to be replicated in one or more selected host cells. Typically, in a cloning vector, this sequence is one that enables the vector to be replicated independently of host chromosomal DNA and includes a replication origin or an autonomously replicating sequence. As used herein, the term "expression vector" means a vector comprising a recombinant polynucleotide that includes an expression regulatory sequence operably linked to a nucleotide sequence to be expressed. The expression vector comprises sufficient cis-acting elements for expression; other elements for expression may be provided by a host cell or an in vitro expression system. Expression vectors include all expression vectors known in the art, such as cosmids, plasmids (e.g., naked or contained in liposomes), and viruses (e.g., lentiviruses, retroviruses, adenoviruses, and adeno-associated viruses).

[0231] As used herein, the term "host cell" refers to a cell that can be used for vector introduction, and is a prokaryotic cell, such as E. coli ( Escherichia coli ) or Bacillus Subtilis ( Bacillus subtilis ), fungal cells, such as yeast cells, or Aspergillus ( AspergillusInsect cells such as S2 fruit fly cells (drosophila cells) or Sf9, or animal cells such as fibroblasts, NS0 cells, Vero cells, Hela cells, COS cells, CHO cells (e.g., CHO-K1, CHO-S, CHO DXB11, ExpiCHO, CHO DG44 or CHO-EBNA cells), ExpiCHO cells, HEK293 cells, Expi293 cells, BHK cells and MDCKII cells, but not limited thereto.

[0232] As used herein, the term "identity" is used to refer to the degree of sequence match between two polypeptides or between two nucleic acids. If the same base or amino acid monomer subunit occupies a position in each of the two sequences being compared (e.g., if adenine occupies a position in each of the two DNA molecules, or if lysine occupies a position in each of the two polypeptides), said molecules are identical at that site. The "percent identity" between two sequences is a function of dividing the number of matching positions shared by the two sequences by the number of positions being compared, and then multiplying by 100. For example, if 6 out of 10 positions in two sequences match, said sequences have 60% identity. For example, the DNA sequences CTGACT and CAGGTT share 50% identity (3 out of a total of 6 positions match). Generally, a comparison is performed when two sequences are aligned to produce maximum identity. Such alignment is, for example, Needleman et al. (1970) J. Mol . Biol .This can be performed using the methods described in 48: 443-453, which is conveniently carried out via a computer program, such as an alignment program (DNAstar, Inc.). The percentage of identity between two amino acid sequences is also determined using the PAM120 weight residue table, gap length penalty 12, and gap penalty 4, as described by E. Meyers and W. Miller (integrated into the ALIGN program (version 2.0)). Comput. Appl . Biosci . The percentage of identity between two amino acid sequences can be determined using the algorithm by Needleman and Wunsch ( , 4: 11-17 (1988)). Additionally, the percentage of identity between two amino acid sequences can be determined using the Blossum 62 matrix or PAM250 matrix, and gap weights 16, 14, 12, 10, 8, 6 or 4, and length weights 1, 2, 3, 4, 5 or 6, integrated into the GAP program of the GCG software package (available at www.gcg.com). J MoI Biol . 48: It can be determined using the algorithm by 444-453 (1970).

[0233] As used herein, the term "conservative substitution" means an amino acid substitution that does not have an adverse effect on or alter the intended properties of a protein / polypeptide containing an amino acid sequence. For example, conservative substitutions may be introduced by standard techniques known in the art, such as 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, for example, to a residue that is physically or functionally similar to the corresponding amino acid residue (e.g., having similar size, shape, charge, chemical properties including the ability to form covalent or hydrogen bonds, etc.). Families of amino acid residues having similar side chains are defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, and histidine), amino acids with acidic side chains (e.g., aspartic acid and glutamic acid), amino acids with uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, and tryptophan), amino acids with nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, and methionine), amino acids with β-branched side chains (e.g., threonine, valine, and isoleucine), and amino acids with aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, and histidine). Therefore, it is desirable to replace the corresponding amino acid residue with another amino acid residue derived from the same side chain family. Methods for identifying conservative amino acid substitutions are well known in the art (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 See 94: 412-417 (1997), which are incorporated herein by reference.

[0234] The designation of the 20 common amino acids mentioned herein follows conventional usage. For example, see Immunology-A Synthesis (2nd Edition, ES. Golub and DR. Gren, Eds., Sinauer Associates, Sunderland, Mass. (1991)), which is incorporated herein by reference. In this disclosure, the terms “polypeptide” and “protein” have the same meaning and are used interchangeably. Furthermore, in this disclosure, amino acids are denoted by one-letter and three-letter abbreviations generally well known in the art. For example, alanine may be denoted as A or Ala.

[0235] As used herein, the term “pharmaceutically acceptable carriers and / or excipients” means carriers and / or excipients that are pharmacologically and / or physiologically suitable for the target and active ingredient, and which are well known in the art (see, e.g., Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995), and include, but are not limited to, pH adjusters, surfactants, adjuvants, ionic strength enhancers, diluents, reagents for maintaining osmotic pressure, absorption retardants, 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 reagents, such as parabens, chlorobutanol, phenol, and sorbic acid. Reagents for maintaining osmotic pressure include, but are not limited to, saccharides, NaCl, and analogs thereof. Absorption retardants include, but are not limited to, monostearates and gelatin. Diluents include, but are not limited to, water, aqueous buffers (e.g., buffered saline), alcohols, polyols (e.g., glycerol), etc. Preservatives include, but are not limited to, various antibacterial and antifungal reagents, such as thiomersal, 2-phenoxyethanol, parabens, chlorobutanol, phenol, and sorbic acid. "Stabilizer" has the meaning generally understood by those skilled in the art and can stabilize the intended activity of the active ingredient in the drug.The above stabilizers include, but are not limited to, sodium glutamate, gelatin, SPGA, saccharides (e.g., sorbitol, mannitol, starch, sucrose, lactose, dextran, or glucose), amino acids (e.g., glutamic acid or glycine), proteins (e.g., dried whey, albumin, or casein), or their degradation products (e.g., lactalbumin hydrolysate).

[0236] As used herein, the term “prevention” means a method performed to prevent or delay the occurrence of a disease, disorder, or symptom (e.g., tumor) in a subject. As used herein, the term “treatment” means a method performed to obtain a beneficial or desired clinical outcome. For the purposes of this disclosure, beneficial or desired clinical outcomes include, but are not limited to, alleviation of symptoms, reduction of disease severity, stabilization of the disease state (i.e., no further deterioration), delay or slowing of disease progression, improvement or alleviation of the disease state, and remission of symptoms (whether partial or complete), regardless of detectability. Additionally, “treatment” may mean an extension of survival time compared to the expected survival time (if treatment were not administered).

[0237] As used herein, the term “subject” means mammals, e.g., primate mammals, e.g., humans. In certain embodiments, said subject (e.g., human) has a tumor or is at risk of having such disease.

[0238] As used herein, the term "effective amount" means an amount sufficient to obtain, or at least partially obtain, the desired effect. For example, an effective amount for preventing a disease (e.g., a tumor) means an amount sufficient to prevent, suppress, or delay the occurrence of the disease (e.g., a tumor); and an effective amount for treating a disease means an amount sufficient to treat or at least partially suppress the disease and its complications in a patient suffering from the disease. Determining such effective amounts is entirely within the capabilities of those skilled in the art. For example, the effective amount for therapeutic use will vary depending on the severity of the disease to be treated, the overall condition of the patient's own immune system, the patient's general circumstances such as age, weight, and gender, the mode of administration of the drug, other concomitant treatments, etc.

[0239] As used herein, the term "effector function" refers to biological activity attributable to the Fc region of an antibody (native-sequence Fc region or amino acid sequence-variant Fc region), and these activities vary depending on the antibody isotype. Examples of antibody effector functions include, but are not limited to, Fc receptor binding affinity, antibody-dependent cell-mediated cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), antibody-dependent cell phagocytosis (ADCP), down-regulation of cell surface receptors (e.g., B cell receptors), B cell activation, cytokine secretion, and the half-life / clearance of antibodies and antigen-antibody complexes. Methods for modifying the effector function of an antibody, for example, by introducing a mutation into the Fc region, are known in the art.

[0240] As used herein, the term "antibody-dependent cell-mediated cytotoxicity (ADCC)" refers to a form of cytotoxicity in which Ig binds to the Fc receptor (FcR) present on cytotoxic cells (e.g., natural killer (NK) cells, neutrophils, or macrophages), causing these cytotoxic effector cells to specifically bind to antigen-attached target cells and subsequently secrete cytotoxins to kill the target cells.

[0241] As used herein, the term "complement-dependent cytotoxicity (CDC)" refers to a complement-mediated cytotoxic effect in which a specific antibody binds to a corresponding antigen on the cell membrane surface to form a complex, which activates the classical complement pathway; and the membrane-attacking complex formed thereby exerts a lytic effect on target cells.

[0242] In this document, "combination" includes regimens that may be administered separately, e.g., regimens that may be formulated separately for individual administration (e.g., provided as a kit), and regimens that may be administered together as a single agent (i.e., "combination-formulation"). In certain embodiments, the anti-ADAM9 antibody of this disclosure or its antigen-binding fragment may be administered sequentially. In other embodiments, the anti-ADAM9 antibody or its antigen-binding fragment may be administered simultaneously. The antibody or its antigen-binding fragment of this disclosure may be used in any combination with at least one other (active) agent.

[0243] In such combination therapies, various activators often possess different complementary mechanisms of action, and the combination therapy can produce synergistic effects. Combination therapy includes agents that affect the immune response (e.g., enhancing or activating the response) and agents that affect tumor / cancer cells (e.g., suppressing or killing them). Combination therapy can reduce the likelihood of drug-resistant cancer cell development. Combination therapy can reduce or eliminate side effects associated with one or more agents by lowering the dosage of one or more agents. Such combination therapies may have therapeutic or prophylactic synergistic effects regarding underlying diseases, disorders, or pathological conditions.

[0244] At our institution, ADAM9 positivity is determined by immunohistochemistry and staining intensity evaluation performed by a specialist clinical pathologist.

[0245] The terms "cancer" and "tumor" are used interchangeably and refer to a broad category of diseases characterized by the uncontrolled growth of abnormal cells within the body. Uncontrolled cell division can lead to the formation of malignant tumors or the development of cells that infiltrate adjacent tissues, and can metastasize to distal parts of the body via the lymphatic system or bloodstream. Cancer includes not only benign or malignant cancers but also dormant tumors or micrometastases. Cancer also includes hematological malignancies.

[0246] Specific embodiments of the present disclosure will be described in detail through the following examples. However, those skilled in the art will understand that the following examples are used merely to illustrate the present disclosure and are not intended to limit the scope of the present disclosure. Various purposes and advantageous aspects of the present disclosure will be realized by those skilled in the art in accordance with the accompanying drawings and the following detailed description of preferred embodiments.

[0247] Sequence information

[0248] Sequence information related to the present disclosure is listed in the table below:

[0249] Explanation of the sequence Sequence Number: Explanation of the sequence Amino acid / nucleotide sequence 1 Amino acid sequence of Ab01 VH QAYLQQSGAELVRPGASVKMSCKASGYTFTNYNVHWVKQTPRQGLEWIGAIYPGNGDTSYNQKFKGKATLTVDKSSSTAYMQLSSLTSEDSAVYFCAIGYSYFDYWGQGTTLTVSS 2 Amino acid sequence of Ab01 VL DIVLTQSPATLSVTPGDSVSLSCRASQSISNNLHWYQQKSHESPRLLIKFASQSISGIPSKFSGSGSGTDFTLSINSVETEDFGMYFCQQSYSWYTFGSGTKLEIK 3 Amino acid sequence of Ab01-HZ43 VH QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYNVHWVRQAPGQGLEWIGAIYPGNGDTSYNQKFKGRVTMTVDTSTSTVYMELSSLRSEDTAVYYCARGYSYFDYWGQGTLVTVSS 4 Amino acid sequence of Ab01-HZ43 VL DIVLTQSPATLSVSPGERATLSCRASQSISNNLHWYQQKPGQAPRLLIKFASQSISGIPARFSGSGSGTEFTLTISSVQSEDFAVYYCQQSYSWYTFGQGTKLEIK 5 Amino acid sequence of Ab01-HZ43 / Ab02-HZ42 / Ab03-HZ43 / Ab04-HZ81 CH ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 6 Amino acid sequence of Ab01-HZ43 / Ab02-HZ42 / Ab03-HZ43 / Ab04-HZ81 CL RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 7 Amino acid sequence of Ab02 VH QVQLQQPGAELVRPGTSVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGLIDPSDTYTNYNQNFKGKATLTVDTSSRIVYMQLSSLTSEDSAVYYCARGDYGNNVGYFDVWGAGTTVTVSP 8 Amino acid sequence of Ab02 VL DIQMIQSPASLSASVGETVTITCRASENIDSYLAWYQQKQGKSPQLLVYVATLLADGVPSRFSGSGSGTQYSLKINSLQSEDVARYYCQHYYITPWTFGGGTKLEIK 9 Amino acid sequence of Ab02-HZ42 VH QVQLVQSGAEVVKPGASVKLSCKASGYTFTNYWMHWVRQRPGQGLEWIGLIDASDSYINYNAKFQGRVTITVDTSTSTAYMELSSLRSEDTAVYYCARGDYGNNVGYFDVWGQGTLVTVSS 10 Amino acid sequence of Ab02-HZ42 VL DIQMTQSPSSLSASVGQRVTITCRASENIDRYLAWYQQKQGKAPKLLVYVATLLASGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQHYYITPWTFGQGTKLEIK 11 Amino acid sequence of Ab01 / Ab01-HZ43 / Ab02-HZ42 Chothia CDR H1 GYTFTNY 12 Amino acid sequence of Ab01 / Ab01-HZ43 Chothia CDR H2 YPGNGD 13 Amino acid sequence of Ab01 / Ab01-HZ43 Chothia / Abm / Kabat CDR H3 GYSYFDY 14 Amino acid sequence of Ab01 / Ab01-HZ43 Chothia / Abm / Kabat CDR L1 RASQSISNNLH 15 Amino acid sequence of Ab01 / Ab01-HZ43 Chothia / Abm / Kabat CDR L2 FASQSIS 16 Amino acid sequence of Ab01 / Ab01-HZ43 Chothia / Abm / Kabat / IMGT CDR L3 QQSYSWYT 17 Amino acid sequence of Ab01 / Ab01-HZ43 Abm CDR H1 GYTFTNYNVH 18 Amino acid sequence of Ab01 / Ab01-HZ43 Abm CDR H2 AIYPGNGDTS 19 Amino acid sequence of Ab01 / Ab01-HZ43 Kabat CDR H1 NYNVH 20 Amino acid sequence of Ab01 / Ab01-HZ43 Kabat CDR H2 AIYPGNGDTSYNQKFKG 21 Amino acid sequence of Ab01 / Ab01-HZ43 IMGT CDR H1 GYTFTNYN 22 Amino acid sequence of Ab01 / Ab01-HZ43 IMGT CDR H2 IYPGNGDT 23 Amino acid sequence of Ab01-HZ43 IMGT CDR H3 ARGYSYFDY 24 Amino acid sequence of Ab01 / Ab01-HZ43 IMGT CDR L1 QSISNN 25 Amino acid sequence of Ab01 / Ab01-HZ43 IMGT CDR L2 FAS 26 Amino acid sequence of Ab02-HZ42 Chothia CDR H2 DASDSY 27 Amino acid sequence of Ab02 / Ab02-HZ42 Chothia / Abm / Kabat CDR H3 GDYGNNVGYFDV 28 Amino acid sequence of Ab02-HZ42 Chothia / Abm / Kabat CDR L1 RASENIDRYLA 29 Amino acid sequence of Ab02-HZ42 Chothia / Abm / Kabat CDR L2 VATLLAS 30 Amino acid sequence of Ab02 / Ab02-HZ42 Chothia / Abm / Kabat / IMGT CDR L3 QHYYITPWT 31 Amino acid sequence of Ab02-HZ42 Abm CDR H1 GYTFTNYWMH 32 Amino acid sequence of Ab02-HZ42 Abm CDR H2 LIDASDSYIN 33 Amino acid sequence of Ab02-HZ42 Kabat CDR H1 NYWMH 34 Amino acid sequence of Ab02-HZ42 Kabat CDR H2 LIDASDSYINYNAKFQG 35 Amino acid sequence of Ab02-HZ42 IMGT CDR H1 GYTFTNYW 36 Amino acid sequence of Ab02-HZ42 IMGT CDR H2 IDASDSYI 37 Amino acid sequence of Ab02 / Ab02-HZ42 IMGT CDR H3 ARGDYGNNVGYFDV 38 Amino acid sequence of Ab02-HZ42 IMGT CDR L1 ENIDRY 39 Amino acid sequence of Ab02 / Ab02-HZ42 IMGT CDR L2 VA 40 Amino acid sequence of Ab01-HZ43 HC QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYNVHWVRQAPGQGLEWIGAIYPGNGDTSYNQKFKGRVTMTVDTSTSTVYMELSSLRSEDTAVYYCARGYSYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 41 Ab01-HZ43 LC의 아미노산 서열 DIVLTQSPATLSVSPGERATLSCRASQSISNNLHWYQQKPGQAPRLLIKFASQSISGIPARFSGSGSGTEFTLTISSVQSEDFAVYYCQQSYSWYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 42 Ab02-HZ42 HC의 아미노산 서열 QVQLVQSGAEVVKPGASVKLSCKASGYTFTNYWMHWVRQRPGQGLEWIGLIDASDSYINYNAKFQGRVTITVDTSTSTAYMELSSLRSEDTAVYYCARGDYGNNVGYFDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 43 Ab02-HZ42 LC의 아미노산 서열 DIQMTQSPSSLSASVGQRVTITCRASENIDRYLAWYQQKQGKAPKLLVYVATLLASGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQHYYITPWTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 44 Ab01-HZ43 VH의 뉴클레오티드 서열 CAAGTACAACTCGTGCAAAGCGGCGCAGAAGTAAAGAAGCCTGGTGCCTCTGTGAAAGTGTCATGCAAGGCCTCCGGTTACACTTTCACCAACTACAACGTACATTGGGTTCGGCAGGCTCCAGGACAAGGACTGGAATGGATCGGAGCCATTTACCCCGGAAACGGGGATACCTCCTACAATCAGAAGTTCAAAGGTAGGGTGACTATGACCGTCGACACCTCTACGAGCACGGTGTATATGGAGTTGTCATCTCTGCGGAGCGAAGATACTGCTGTGTATTATTGTGCAAGGGGATACTCTTACTTCGATTACTGGGGGCAAGGAACACTCGTAACGGTGTCCAGC 45 Ab01-HZ43 VL의 뉴클레오티드 서열 GACATCGTCCTTACCCAGTCACCCGCCACTCTTTCCGTAAGCCCTGGAGAGAGAGCTACGCTTTCCTGTAGAGCCTCCCAGTCCATTTCAAACAACCTGCACTGGTATCAACAGAAACCAGGACAGGCCCCTAGATTGCTGATCAAATTCGCTAGCCAATCTATCTCTGGGATACCTGCTCGCTTCAGTGGATCCGGTTCTGGTACAGAGTTCACCCTCACCATCAGCTCTGTCCAGTCAGAAGACTTCGCCGTGTACTACTGCCAGCAGTCATACTCTTGGTACACCTTCGGCCAGGGGACTAAGCTCGAAATCAAG 46 Ab02-HZ42 VH의 뉴클레오티드 서열 CAAGTCCAGCTGGTGCAAAGCGGAGCAGAAGTAGTAAAGCCGGGTGCTAGTGTCAAACTGTCTTGTAAGGCTTCTGGGTACACTTTCACGAATTACTGGATGCATTGGGTCAGGCAGAGACCCGGACAAGGGCTTGAATGGATCGGACTGATTGATGCTAGCGACAGCTACATCAACTACA ACGCTAAGTTTCAGGGGAGGGTTACGATCACCGTCGATACCAGCACCAGCACAGCCTACATGGAGCTCTCTTCATTGAGATCTGAGGATACAGCAGTCTACTACTGTGCCAGAGGCGATTATGGTAACAACGTAGGGTATTTTGACGTCTGGGGTCAGGGTACTCTGGTGACTGTCAGTAGC 47 Nucleotide sequence of Ab02-HZ42 VL GATATCCAGATGACACAGTCCCCGAGTTCCCTCTCCGCTAGCGTAGGACAAAGGGTGACTATTACATGCCGCGCCAGCGAGAATATTGACAGGTATCTCGCCTGGTATCAGCAGAAACAGGGTAAGGCCCCTAAGTTGTTGGTGTACGTTGCCACTCTTC TGGCTTCTGGTGTTCCTAGCAGATTCTCCGGTAGCGGTAGTGGTACCGATTACACTCTGACAATCTCCTCATTGCAGCCGGAAGATTTTGCCACATACTATTGCCAGCACTACTACATAACACCGTGGACCTTTTGGCCAGGGAACCAAATTGGAGATCAAG 48 Amino acid sequence of Ab03 VH QVQLLQPGAELVKPGTSVKLSCKASGYTFTTYWMHWVKQRPGRGLEWIGRFDPNSGSSKYNEKFKSKATLTVDKPSSTAYMQLNSLTSEDSAVYYCATDTYDYFDYWGQGTTLTVSS 49 Amino acid sequence of Ab03 VL ETTVTQSPASLSMAIGEKVTIRCITSTDIDDDMSWYQQKAGEPPKLLVSDGNTLRPGVPSRFSASGYGTDFVFTIENMLSEDVADYYCLQSENLPYTFGGGTKLEIK 50 Amino acid sequence of Ab03-HZ43 VH QVQLVQSGAEVKKPGASVKVSCKASGYTFTTYWMHWVRQAPGQGLEWIGRFDPNSGSSKYNEKFKSRVTLTVDTSISTAYMELSRLRSDDTAVYYCATDTYDYFDYWGQGTLVTVSS 51 Amino acid sequence of Ab03-HZ43 VL ETTLTQSPAFMSATPGDKVNISCITSTDIDDDMSWYQQKPGEPPILLVSDGNTLRPGIPPRFSGSGYGTDFTLTINNMESEDAATYYCLQSENLPYTFGGGTKLEIK 52 Amino acid sequence of Ab04 VH DVQLQESGPGLVKPSQSLSLTCSVTGYSITSGYFWNWIRQFPGNKLEWMGYISYDGSNNYNPSLKSRISITRDTSKNQFFLKLNSVTTEDTATYYCTRSDAAYYDYDVGWGQGTLVTVSA 53 Amino acid sequence of Ab04 VL DIQMTQSPASLSASVGETVTITCRASENIDSYLAWYQQKQGKSPQLLVYAATLLADGVPSRFSGSGSGTQYSLKINSLQSEDVARYYCQHYYITPLTFGAGTKLELK 54 Amino acid sequence of Ab04-HZ81 VH QVQLQESGPGLVKPSQTLSLTCTVSGYSITSGYFWNWIRQFPGKGLEWMGYISYDGSNNYNPSLKSRITISRDTSKNQFSLKLNSVTAADTAVYYCTRSDAAYYDYDVGWGQGTLVTVSS 55 Amino acid sequence of Ab04-HZ81 VL DIQMTQSPSSLSASVGDRVTITCRASENIDSYLAWYQQKPGKAPKLLIYAATLLADGVPSRFSGSGSGTDFTLTINSLQPEDVATYYCQHYYITPLTFGQGTKLEIK 56 Amino acid sequence of Ab03 / Ab03-HZ43 Chothia CDR H1 GYTFTTY 57 Amino acid sequence of Ab03 / Ab03-HZ43 Chothia CDR H2 DPNSGS 58 Amino acid sequence of Ab03 / Ab03-HZ43 Chothia CDR H3 DTYDYFDY 59 Amino acid sequence of Ab03 / Ab03-HZ43 Chothia / Kabat / Abm CDR L1 ITSTDIDDDMS 60 Amino acid sequence of Ab03 / Ab03-HZ43 Chothia / Kabat / Abm CDR L2 DGNTLRP 61 Amino acid sequence of Ab03 / Ab03-HZ43 Chothia / Kabat / Abm CDR L3 LQSENLPYT 62 Amino acid sequence of Ab03 / Ab03-HZ43 Abm / Kabat CDR H1 TYWMH 63 Amino acid sequence of Ab03 / Ab03-HZ43 Abm / Kabat CDR H2 RFDPNSGSSKYNEKFKS 64 Amino acid sequence of Ab03 / Ab03-HZ43 Abm / Kabat CDR H3 DTYDYFDY 65 Amino acid sequence of Ab03 / Ab03-HZ43 IMGT CDR H1 GYTFTTYW 66 Amino acid sequence of Ab03 / Ab03-HZ43 IMGT CDR H2 FDPNSGSS 67 Amino acid sequence of Ab03 / Ab03-HZ43 IMGT CDR H3 ATDTYDYFDY 68 Amino acid sequence of Ab03 / Ab03-HZ43 IMGT CDR L1 TDIDDD 69 Amino acid sequence of Ab03 / Ab03-HZ43 IMGT CDR L2 DGN 70 Amino acid sequence of Ab03 / Ab03-HZ43 IMGT CDR L3 LQSENLPYT 71 Amino acid sequence of Ab04 / Ab04-HZ81 Chothia CDR H1 GYSITSGY 72 Amino acid sequence of Ab04 / Ab04-HZ81 Chothia CDR H2 SYDGS 73 Amino acid sequence of Ab04 / Ab04-HZ81 Chothia / Kabat / Abm CDR H3 SDAAYYDYDVG 74 Amino acid sequence of Ab04 / Ab04-HZ81 Chothia / Kabat / Abm CDR L1 RASENIDSYLA 75 Amino acid sequence of Ab04 / Ab04-HZ81 Chothia / Kabat / Abm CDR L2 AATLLAD 76 Amino acid sequence of Ab04 / Ab04-HZ81 Chothia / Kabat / Abm / IMGT CDR L3 QHYYITPLT 77 Amino acid sequence of Ab04 / Ab04-HZ81 Abm CDR H1 GYSITSGYFWN 78 Amino acid sequence of Ab04 / Ab04-HZ81 Abm CDR H2 YISYDGSNN 79 Amino acid sequence of Ab04 / Ab04-HZ81 Kabat CDR H1 SGYFWN 80 Amino acid sequence of Ab04 / Ab04-HZ81 Kabat CDR H2 YISYDGSNNYNPSLKS 81 Amino acid sequence of Ab04 / Ab04-HZ81 IMGT CDR H1 GYSITSGYF 82 Amino acid sequence of Ab04 / Ab04-HZ81 IMGT CDR H2 ISYDGSN 83 Amino acid sequence of Ab04 / Ab04-HZ81 IMGT CDR H3 TRSDAAYYDYDVG 84 Amino acid sequence of Ab04 / Ab04-HZ81 IMGT CDR L1 ENIDSY 85 Amino acid sequence of Ab04 / Ab04-HZ81 IMGT CDR L2 AAT 86 Amino acid sequence of Ab03-HZ43 HC QVQLVQSGAEVKKPGASVKVSCKASGYTFTTYWMHWVRQAPGQGLEWIGRFDPNSGSSKYNEKFKSRVTLTVDTSISTAYMELSRLRSDDTAVYYCATDTYDYFDYWGQGT LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 87 Amino acid sequence of Ab03-HZ43 LC ETTLTQSPAFMSATPGDKVNISCITSTDIDDDMSWYQQKPGEPPILLVSDGNTLRPGIPPRFSGSGYGTDFTLTINNMESEDAATYYCLQSENLPYTFGGGTKLEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 88 Amino acid sequence of Ab04-HZ81 HC QVQLQESGPGLVKPSQTLSLTCTVSGYSITSGYFWNWIRQFPGKGLEWMGYISYDGSNNYNPSLKSRITISRDTSKNQFSLKLNSVTAADTAVYYCTRSDAAYYDYDVGWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 89 Ab04-HZ81 LC의 아미노산 서열 DIQMTQSPSSLSASVGDRVTITCRASENIDSYLAWYQQKPGKAPKLLIYAATLLADGVPSRFSGSGSGTDFTLTINSLQPEDVATYYCQHYYITPLTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 90 Ab03-HZ43 VH의 뉴클레오티드 서열 CAAGTGCAACTGGTGCAGAGTGGTGCCGAAGTGAAGAAACCAGGGGCCTCCGTCAAGGTCAGCTGTAAGGCATCCGGCTACACATTCACAACTTATTGGATGCATTGGGTCAGACAAGCTCCTGGGCAGGGACTGGAATGGATAGGTCGGTTCGATCCAAACTCCGGATCCAGCAAGTACAACGAGAAGTTCAAGAGCCGGGTGACACTTACTGTCGACACCTCTATATCCACAGCCTATATGGAGCTCTCACGCCTGAGATCCGATGATACGGCCGTCTACTACTGCGCTACAGACACCTATGATTACTTCGACTACTGGGGACAGGGGACACTGGTCACTGTGTCATCA 91 Ab03-HZ43 VL의 뉴클레오티드 서열 GAAACCACACTTACACAGTCACCTGCATTTATGTCAGCAACCCCAGGCGACAAAGTTAACATCAGCTGTATTACTAGCACCGACATCGACGATGACATGTCCTGGTACCAGCAAAAGCCTGGTGAGCCTCCTATCCTCTTGGTTTCAGACGGCAATACTCTCAGACCAGGTATCCCACCTCGATTTTCCGGCTCTGGTTATGGGACAGATTTCACCCTGACGATCAACAACATGGAATCTGAAGACGCTGCCACTTACTACTGTCTGCAGAGTGAGAATTTGCCTTACACCTTTGGTGGAGGAACTAAACTGGAAATCAAA 92 Ab04-HZ81 VH의 뉴클레오티드 서열 CAAGTACAGTTGCAAGAAAGCGGCCCTGGCCTGGTCAAGCCCTCTCAGACCCTGTCCCTGACCTGTACAGTCAGCGGATATTCCATAACCTCCGGGTACTTTTGGAACTGGATTAGGCAATTTCCAGGCAAAGGCCTGGAATGGATGGGCTATATCTCATACGACGGTAGCAACAATTAT AATCCCAGTCTGAAAAGCCGGATTACCATCAGTCGGGACACTTCTAAGAACCAGTTCTCCCTCAAGTTGAATAGTGTCACCGCAGCCGATACAGCTGTCTACTATTGTACTAGATCCGATGCCGCCTATTATGATTACGACGTTGGCTGGGGTCAAGGAACTCTGGTGACGGTAAGTTCT 93 Nucleotide sequence of Ab04-HZ81 VL GACATCCAGATGACCCAAAGCCCCTCAAGCCTTAGCGCCTCCGTTGGCGACAGAGTGACAATCACATGCCGCGCCTCTGAGAACATCGACTCCTATCTCGCATGGTATCAGCAAAAGCCAGGTAAGGCTCCCAAGCTTCTGATCTATGCTGCTACGCTGT TGGCTGATGGAGTGCCTAGCCGGTTTAGTGGAAGTGGTTCTGGCACAGATTTCACACTGACCATAAACAGCCTGCAACCTGAAGATGTAGCCACTTATTACTGCCAGCACTACTATATCACGCCATTGACCTTCGGGCAGGGCACTAAGCTTGGAAATCAAA 94 Amino acid sequence of Ab01 IMGT CDR H3 AIGYSYFDY 95 Amino acid sequence of Ab02 Chothia CDR H1 GYTFTSY 96 Amino acid sequence of Ab02 Chothia CDR H2 DPSDTY 97 Amino acid sequence of Ab02 Chothia / Abm / Kabat CDR L1 RASENIDSYLA 98 Amino acid sequence of Ab02 Chothia / Abm / Kabat CDR L2 VATLLAD 99 Amino acid sequence of Ab02 Abm CDR H1 GYTFTSYWMH 100 Amino acid sequence of Ab02 Abm CDR H2 LIDPSDTYTN 101 Amino acid sequence of Ab02 Kabat CDR H1 SYWMH 102 Amino acid sequence of Ab02 Kabat CDR H2 LIDPSDTYTNYNQNFKG 103 Amino acid sequence of Ab02 IMGT CDR H1 GYTFTSYW 104 Amino acid sequence of Ab02 IMGT CDR H2 IDPSDTYT 105 Amino acid sequence of Ab02 IMGT CDR L1 ENIDSY

[0250] Specific examples details

[0251] The present disclosure is described with reference to the following examples, which are illustrative of (but not limiting to) the present disclosure.

[0252] Unless otherwise specified, the molecular biology experimental methods and immunoassays used in this disclosure are essentially J. Sambrook et al. , Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press, 1989, and FM Ausubel. et al. This is performed by referring to the method described in Short Protocols In Molecular Biology, 3rd Edition, John Wiley & Sons, Inc., 1995. Those skilled in the art know that the embodiments are for illustrative purposes only and are not intended to limit the claimed scope of the disclosure.

[0253] Examples 1: ADAM9 Preparation of antigen and control antibody proteins

[0254] 1.1 Preparation of ADAM9 Antigen

[0255] The extracellular domain sequences of human ADAM9 (UniProt: Q13443) and monkey ADAM9 (UniProt: A0A2K5X4X8) were gene-synthesized using GenScript, and then constructed into a pTT5 vector by introducing six histidines at the C-terminus. Recombinant plasmids were extracted. After confirming that the sequencing was accurate, the plasmids were transiently transfected into HEK293 cells and expressed transiently. On day 6, the cell supernatant was collected, and human ADAM9 protein and monkey ADAM9 protein were obtained through purification.

[0256] 1.2 Expression of ADAM9 control antibody

[0257] The ADAM9 control antibody was derived from hMAB-A(2I .2) of patent CN 201980049672. After codon optimization by GenScript, the heavy chain and light chain constant regions of the antibody were as presented in SEQ ID NO: 5 and SEQ ID NO: 6 of the said patent application, respectively. The nucleotide sequences of the antibody heavy chain and light chain were synthesized and cloned into the pTT5 vector. Then, the pTT5 plasmids corresponding to the nucleotides of the antibody heavy chain and light chain were co-transfected into CHOS cells. After collecting the cell supernatant by centrifugation, the antibodies in the supernatant were purified using Protein A (MabSelect SuRe, GE) to obtain the control antibody protein.

[0258] 1.3 Preparation of ADAM9 Overexpressing Cell Line

[0259] Based on the full-length proteins of human ADAM9 (NP_003807.1), monkey ADAM9 (XP_045253681.1), rat ADAM9 (NP_001014772.2), and mouse ADAM9 (NP_001257925.1), codon optimization and gene synthesis were commissioned to Nanjing GenScript Biotech Co., Ltd. The above genes were constructed into the lentiviral vector pLVX-IRES-puro, followed by viral packaging and infection of BaF3 cells. After verification through pressure screening and flow cytometry, overexpression cell lines stably expressing human, monkey, rat, and mouse ADAM9 were obtained.

[0260] Examples 2: Anti-human ADAM9 Monoclonal Antibody production

[0261] 2.1 Immunization of mice

[0262] Wild-type mice were immunized with the human ADAM9 protein prepared in Section 1.1. Specifically, the first immunization was performed using Freund's complete adjuvant, and subsequent booster immunizations were performed at 1-week intervals using Freund's incomplete adjuvant. During immunization, serum titers of the anti-ADAM9 antibody were monitored every two weeks via protein ELISA and flow cytometry. The specific procedure for the ELISA is as follows: Human ADAM9-his was diluted to 1 μg / ml in CBS-coated solution, added at a rate of 100 μl per well, and incubated overnight at 4°C. The sample was washed once with 300 μl of PBST, and blocked by adding 100 μl of PBS + 2% BSA at 37°C for 2 hours. The blocking solution was discarded. Serum was diluted in a PBS + 2% BSA solution, starting with a 100-fold dilution and followed by 3-fold gradient dilutions, and then incubated at 37°C for 2 hours. Washing was performed three times with 300 μl of PBST. HRP-goat-anti-mouse IgG was diluted in PBS (2% BSA) at a ratio of 1:10,000. 100 μl of the diluted solution was added to a well plate and then incubated at 37°C for 1 hour. Washing was performed five times with 300 μl of PBST. 80 μl of TMB (Huzhou InnoReagents Co., Ltd.) was added. After color development, 40 μl of 2N H2SO4 was added to terminate the reaction, and the absorbance was read at 450 nm using a microplate reader. Flow cytometry analysis: HCT116 cells were collected and washed three times with PBS.Mouse serum was diluted in PBS + 1% BSA solution in 3-fold increments, starting with a 50-fold dilution, to obtain a total of 11 concentrations. The volume per well was 50 μL. Simultaneously, blank cell wells and negative mouse serum control wells were set up. NCI-H226 cells were in PBS + 1% BSA solution at a rate of 1 × 10⁶. 5 The cells were resuspended at a density of cells / well. The cell suspension was individually added to each well at a density of 50 μL / well. After uniform mixing, the cells were incubated at 4°C for 1 hour. Washing with PBS was performed three times. The APC anti-mouse IgG Fc antibody was diluted in PBS + 1% BSA solution (1 μL / well, 100 μL). The cells were resuspended and then incubated at 4°C for 0.5 hours. Washing with PBS was performed three times. The cells were resuspended in 300 μL of PBS and analyzed using a flow cytometer. Mice best suited for hybridoma fusion were selected based on protein-binding activity and cellular titer.

[0263] 2.2 Hybridoma Fusion and Screening

[0264] Seven days after hybridoma fusion, supernatant screening was performed using ELISA. Specifically, monkey ADAM9 protein was diluted to 1 μg / ml in the coating solution, added at a rate of 100 μl / well, and coated overnight at 4°C. One wash was performed with 300 μL of PBST. 100 μl of PBS + 2% BSA was added to each well and incubated at 37°C for 1 hour. Then, 20 μl of the hybridoma supernatant was added directly to the microplate and incubated at 37°C for 2 hours. The solution was discarded, and the microplate was washed three times with 320 μl / well of PBST. The microplate was drained. 100 μL of HRP-goat anti-mouse IgG (Thermo Fisher) diluted to a ratio of 1:10,000 was added to each well and incubated at 37°C for 1 hour. The solution was discarded, and the microplate was washed 5 times with PBST. The microplate was drained. 80 μL of TMB (Huzhou InnoReagents Co., Ltd.) was added to each well in the dark to induce color development. Then, 40 μL of 2N H2SO4 was added to terminate the reaction, and the absorbance was read at 450 nm using a microplate reader. Clones with a signal value 5-fold higher than that of the negative control were further verified by ELISA using human ADAM9 protein and flow cytometry using overexpression cells prepared in Section 1.3. Positive clones were subcloned. Monoclonal clones were screened using the above method, and scale-up culture was performed after selecting the optimal monoclonal clone.

[0265] Examples 3: Anti-human ADAM9 Murin Evaluation of antibodies

[0266] 3.1 Evaluation of Binding Activity of Anti-Human ADAM9 Murine Antibody

[0267] All selected monoclonal clones were scaled up for serum-free culture. 5–10 ml of culture supernatant was affinity-purified using Protein-A beads. The eluted product was neutralized with Tris solution. Antibody protein concentrations were quantified using Nanodrop and then used for candidate evaluation.

[0268] Adherent lung squamous cell carcinoma cells EBC-1 (Nanjing Cobioer) were digested with StemPro™ Accutase (Gibco) solution and counted. An appropriate number of cells were obtained, washed twice with 1× PBS, and resuspended in 1% BSA solution. Then, the cells were transferred to a 96-well V-bottom plate at a rate of 50 μL / well. The candidate antibody was diluted in 1% BSA solution in a 3-fold gradient starting at 10 μg / mL. Next, 50 μL of the diluted antibody was added to the V-bottom plate containing the cells, and the mixture was incubated at 4°C for 45 minutes. The cells were washed twice with PBS. Then, 50 μL of the diluted secondary antibody was added to each well. After uniform mixing, the mixture was incubated at 4°C for 30 minutes. Cells were washed twice with PBS. Then, cells were resuspended in 400 μL of PBS and analyzed by flow cytometry. Data processing: Median PE values ​​were extracted and entered into GraphPad Prism 6 software to calculate EC 50 The results were calculated. The results are shown in Figure 1 and Table 2. Antibodies Ab01 to Ab04 all exhibited high affinity levels of 10E-10.

[0269]

[0270] 3.2 Sequencing of Anti-Human ADAM9 Murine Antibody and Production of Chimeric Antibody

[0271] Hybridoma cells were cultured to approximately 8,000 cells and then lysed. The first strand of cDNA was synthesized using a cDNA reverse transcription kit (Thermo Fisher). The VH and VK genes were amplified from the cDNA via PCR using primers and then sequenced. As shown in Table 3, the variable region sequences of the anti-human ADAM9 antibody were obtained.

[0272]

[0273] Examples 4: Anti-human ADAM9 Evaluation of antibody humanization

[0274] 4.1 Humanization and Expression of Anti-Human ADAM9 Antibodies

[0275] Humanization of murine antibodies Ab01, Ab02, Ab03, and Ab04 was performed using the CDR grafting method for antibody humanization. Briefly, humanization involves the following steps: the amino acid sequences of the murine monoclonal antibodies were aligned with the amino acid sequences of human germline antibodies to identify sequences with high homology and excellent physicochemical properties, which were then used as human germline framework sequences. HLA-DR affinities were analyzed and investigated to select human germline framework sequences with low affinity. Subsequently, six CDRs of the murine antibodies were grafted correspondingly to the selected heavy and light chain framework sequences.

[0276] In addition, computer simulation techniques were utilized to analyze the variable region and surrounding framework amino acid sequences through molecular docking, and their spatial stereoscopic binding modes were investigated. By calculating electrostatic, van der Waals, hydrophilic-hydrophobic, and entropy values, key amino acids were analyzed in the amino acid sequences of murine antibodies that interact with the ADAM9 protein and maintain their spatial structure. These murine-derived amino acids were retained in the grafted antibodies. Specifically, a series of back mutations were performed on amino acid residues in the FR region of the humanized templates to ensure that the humanized antibodies retained the antigen-binding ability of the murine antibodies to the maximum extent.

[0277] Codon optimization for the humanized antibody was commissioned to Nanjing GenScript Biotech Co., Ltd., and the synthesized cDNA was ligated into the expression plasmid pTT5. The expression plasmids for the heavy and light chains of the humanized antibody were co-transfected into CHO-S cells. After 7 days of expression, the supernatant was collected by centrifugation. The recombinant antibodies in the supernatant were purified using Protein A (MabSelect SuRe, GE) to obtain the anti-human ADAM9 chimeric antibody and the humanized antibody. The sequences of the variable region and CDRs of the humanized antibody are shown in Table 4.

[0278]

[0279] 4.2 Detection of Protein Affinity of Anti-Human ADAM9 Humanized Antibodies

[0280] The dynamic affinities of chimeric antibodies Ab01-CH and Ab02-CH and humanized antibodies Ab01-HZ43, Ab02-HZ42, Ab03-HZ43 and Ab04-HZ81 against human and monkey ADAM9-His were detected using ForteBio (Pall Life Sciences).

[0281] The specific method is as follows: The test antibody was diluted to 5 μg / ml in PBST (0.02% Tween 20), and human and monkey ADAM9-His proteins were serially diluted to 200 nM, 100 nM, 50 nM, 25 nM, 12.50 nM, 6.25 nM, 3.125 nM, and 0 nM. Subsequently, the test antibody was captured for 60 seconds using Protein A Senso (Pall Life Sciences) in PBST (0.02% Tween-20) solution, followed by binding to the human or monkey ADAM9-His protein for 60 seconds and dissociation for 180 seconds. The results were obtained. The obtained results were entered into Data Analysis 11.0 software. A 1:1 model with global fitting applied was selected in the software to analyze the results, thereby obtaining the binding rate, dissociation rate, and affinity constant.

[0282] The results are shown in Table 5. The dissociation rate constants (kdis) of Ab04-HZ81, Ab01-HZ43, and Ab02-HZ42 for human ADAM9-His were lower than those of the positive control hMAB-A (2I.2), indicating high kinetic affinity. The dissociation constant and dissociation rate constant of Ab04-HZ81 for monkey ADAM9-His reached the same level as those of the positive control hMAB-A (2I.2), indicating high kinetic affinity.

[0283]

[0284] 4.3 Detection of Cell Affinity of Anti-Human ADAM9 Humanized Antibodies

[0285] Adherent lung squamous cell carcinoma cells EBC-1 (Nanjing Cobioer) and human breast cancer cells HCC1806 (Nanjing Cobioer) were digested with StemPro™ Accutase (Gibco) solution. After counting, an appropriate number of cells were obtained, washed twice with 1× PBS, and resuspended in 1% BSA solution. Then, the cells were transferred to 96-well V-bottom plates at a rate of 50 μL / well. The candidate antibody was diluted in 1% BSA in a 3-fold gradient starting at 10 μg / mL. Next, 50 μL of the diluted antibody was added to the V-bottom plates containing the cells, and the mixture was incubated at 4°C for 45 minutes. The cells were washed twice with PBS. Then, 50 μL of the diluted secondary antibody was added to each well. After uniform mixing, the mixture was incubated at 4°C for 30 minutes. The cells were washed twice with PBS. Then, the cells were resuspended in 400 μL of PBS and analyzed by flow cytometry. Data processing: After extracting the median PE value, it was entered into GraphPad Prism 6 software to calculate EC 50 The results were calculated. The results are shown in Figures 2 and 3 and Table 6. The antibodies Ab01-CH, Ab01-HZ43, Ab02-CH, Ab02-HZ42, Ab03-HZ43, and Ab04-HZ81 all showed high affinity.

[0286]

[0287] 4.4 Detection of Endocytotic Activity of Anti-Human ADAM9 Humanized Antibodies

[0288] The endocytic activity of humanized anti-human ADAM9 antibodies in human lung squamous cell carcinoma EBC-1 cells (Nanjing Cobioer) was detected using a flow cytometer (Thermo, model: Attune NxT). The specific procedure was as follows: EBC-1 cells were plated at 10,000 cells / well and incubated overnight. The test antibodies were diluted in culture medium in 2-fold series dilutions, starting at 0.3 μg / ml. PHrodo was diluted in culture medium to 12 μg / ml. The series-diluted antibodies were uniformly mixed with PHrodo in a 1:1 ratio, and the mixture was incubated in the dark at room temperature for 30 minutes. Subsequently, 50 μL of fresh complete medium and 50 μL of the labeled antibodies were added to the cells, and co-incubated for 24 hours at 37°C and 5% CO2. The above cells were washed with PBS and analyzed by flow cytometry. Data processing: After extracting fluorescence signal values, they were input into GraphPad Prism 6 software to perform EC 50 The results were calculated. The results are shown in Figure 4 and Table 7. The chimeric antibody Ab01-CH and its corresponding humanized antibody Ab01-HZ43 exhibited the most potent endocytotic activity, while Ab02-HZ42, Ab03-HZ43, and Ab04-HZ81 exhibited endocytotic activity similar to that of the control antibody hMAB-A (2I.2).

[0289]

[0290] 4.5 Detection of Hydrophobicity of Anti-Human ADAM9 Humanized Antibodies

[0291] Hydrophobicity was detected using hydrophobic interaction chromatography, with the following conditions: Chromatography column: TOSOH TSKgel Butyl-NPR 100*4.6 mm, 2.5 μm; Mobile phase A: 25 mmol / L disodium hydrogen phosphate, pH 7.0, 25% isopropanol; Mobile phase B: 1.5 mol / L ammonium sulfate. Samples were diluted to 1 mg / ml using 0.75 mol / L ammonium sulfate. 40 μg of each sample was injected. Gradient elution was performed, in which hydrophilic samples were eluted first and hydrophobic samples were eluted subsequently. The hydrophilicity or hydrophobicity of the samples was determined based on their retention times. The results are shown in Table 8. Ab01-HZ43, Ab02-HZ42, Ab03-HZ43, and Ab04-HZ81 all exhibited good hydrophilicity, which was superior to the control antibody hMAB-A (2I.2).

[0292]

[0293] 4.6 Detection of Specificity of Humanized Anti-Human ADAM9 Antibodies

[0294] To detect the specificity of the humanized anti-human ADAM9 antibody, human ADAM9, ADAM12 (Sino Biological), and ADAM15 (Sino Biological) proteins were diluted to 1 μg / ml in CBS coating solution. 100 μL / well of the diluted solution was coated onto a 96-well microplate and incubated overnight at 4°C. The microplate was washed once with PBS and drained. Then, 100 μL of 2% BSA was added to each well, followed by incubation at 37°C for 2 hours. Afterward, the blocking solution was removed. The test antibody was diluted to 10 μg / ml in 2% BSA. Then, 100 μL of the diluted antibody was added to each well of the microplate, followed by incubation at 37°C for 2 hours. The plate was washed three times with PBST (containing 0.2% Tween-20) and drained. Next, 100 μL of HRP goat anti-human IgG (H+L) (Jackson) was added to each well and incubated at 37°C for 1 hour. The plate was washed 5 times with PBST (0.2% Tween-20) and drained. Then, 100 μL of TMB chromogenic substrate (Huzhou InnoReagents) was added to each well. After reacting at room temperature for 3-5 minutes, 50 μL of 2N H2SO4 was added to each well to terminate the chromogenic reaction, and the absorbance was read at OD450 nm using a microplate reader.

[0295] The results are as shown in Figure 5. All tested ADAM9 antibodies bound only to the ADAM9 protein and did not bind to the ADAM12 or ADAM15 proteins.

[0296] Although specific embodiments of the present disclosure have been described in detail, those skilled in the art will understand that various modifications and changes to the details may be made in accordance with all the teachings disclosed, and that such changes are included within the scope of protection of the present disclosure. The entire scope of the present disclosure is defined by the claims and equivalents appended herein.

Claims

Claim 1 As an antibody or its antigen-binding fragment that specifically binds to ADAM9, said antibody or its antigen-binding fragment comprises the following complementarity determining regions (CDRs): (a) CDR-H1, CDR-H2, and CDR-H3 contained in the heavy chain variable region (VH) presented in SEQ ID NO: 54 or 52; and / or CDR-L1, CDR-L2, and CDR-L3 contained in the light chain variable region (VL) presented in SEQ ID NO: 55 or 53; (b) CDR-H1, CDR-H2, and CDR-H3 contained in the heavy chain variable region (VH) presented in SEQ ID NO: 3 or 1; and / or CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) presented in SEQ ID NO: 4 or 2; (c) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) presented in SEQ ID NO: 9 or 7; and / or CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) presented in SEQ ID NO: 10 or 8; (d) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) presented in SEQ ID NO: 50 or 48; and / or CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) presented in SEQ ID NO: 51 or 49; or (e) the heavy chain variable region (VH) and / or light chain variable region (VL) comprises a mutation in at least one CDR compared to any one of the heavy chain variable region and / or light chain variable region of (a) to (d), said mutation being a substitution, deletion, or addition of one or several amino acids (e.g., a substitution, deletion, or addition of 1, 2, or 3 amino acids);Preferably, the substitution comprises CDR-H1, CDR-H2, and CDR-H3 contained in the heavy chain variable region (VH) described above, and / or CDR-L1, CDR-L2, and CDR-L3 contained in the light chain variable region (VL) described above, wherein the substitution is a conservative substitution, and preferably, the CDRs are antibodies or their antigen-binding fragments defined according to the IMGT, Kabat, Chothia, or AbM numbering system. Claim 2 In claim 1, the antibody or its antigen-binding fragment comprises: (1) a heavy chain variable region (VH) and / or a light chain variable region (VL) as described below, wherein CDRs are defined by the IMGT numbering system: (1a) a heavy chain variable region (VH) comprising three CDRs: CDR-H1 having the sequence of SEQ ID NO: 81 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 82 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 83 or a variant thereof; and / or, a light chain variable region (VL) comprising three CDRs: CDR-L1 having the sequence of SEQ ID NO: 84 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 85 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 76 or a variant thereof; (1b-i) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 21 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 22 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 23 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 24 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 25 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 16 or a variant thereof; (1b-ii) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 21 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 22 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 94 or a variant thereof; and / or, a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 24 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 25 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 16 or a variant thereof;(1c-i) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 35 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 36 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 37 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 38 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 39 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 30 or a variant thereof; (1c-ii) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 103 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 104 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 37 or a variant thereof; and / or, a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 105 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 39 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 30 or a variant thereof; or (1d) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 65 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 66 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 67 or a variant thereof; and / or, a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 68 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 69 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 70 or a variant thereof;Or, (2) heavy chain variable regions (VH) and / or light chain variable regions (VL) described below, wherein CDRs are defined by the Chothia numbering system: (2a) a heavy chain variable region (VH) comprising three of the following CDRs: CDR-H1 having the sequence of SEQ ID NO: 71 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 72 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 73 or a variant thereof; and / or, a light chain variable region (VL) comprising three of the following CDRs: CDR-L1 having the sequence of SEQ ID NO: 74 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 75 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 76 or a variant thereof; (2b) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 11 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 12 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 13 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 14 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 15 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 16 or a variant thereof; (2c-i) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 11 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 26 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 27 or a variant thereof; and / or, a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 28 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 29 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 30 or a variant thereof;(2c-ii) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 95 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 96 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 27 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 97 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 98 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 30 or a variant thereof; or (2d) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 56 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 57 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 58 or a variant thereof; and / or, a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 59 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 60 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 61 or a variant thereof; or, (3) a heavy chain variable region (VH) and / or a light chain variable region (VL) described below, where the CDRs are defined by the Kabat numbering system: (3a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 79 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 80 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 73 or a variant thereof; and / or, a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 74 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 75 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 76 or a variant thereof;(3b) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 19 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 20 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 13 or a variant thereof; and / or, Light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 14 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 15 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 16 or a variant thereof; (3c-i) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 33 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 34 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 27 or a variant thereof; and / or, a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 28 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 29 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 30 or a variant thereof; (3c-ii) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 101 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 102 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 27 or a variant thereof; and / or, a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 97 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 98 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 30 or a variant thereof; or (3d) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 62 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 63 or a variant thereof;and CDR-H3 having the sequence of SEQ ID NO: 64 or a variant thereof; and / or, a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 59 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 60 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 61 or a variant thereof; or, (4) a heavy chain variable region (VH) and / or a light chain variable region (VL) described below, where the CDRs are defined by the AbM numbering system: (4a) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 77 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 78 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 73 or a variant thereof; and / or, a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 74 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 75 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 76 or a variant thereof; (4b) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 17 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 18 or a variant thereof; and CDR-H3 having the 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 sequence of SEQ ID NO: 14 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 15 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 16 or a variant thereof; (4c-i) Heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 31 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 32 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 27 or a variant thereof;and / or, a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 28 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 29 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 30 or a variant thereof; (4c-ii) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 99 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 100 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 27 or a variant thereof; and / or, a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 97 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 98 or a variant thereof; and CDR-L3 having the sequence of SEQ ID NO: 30 or a variant thereof; or (4d) a heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 having the sequence of SEQ ID NO: 62 or a variant thereof; CDR-H2 having the sequence of SEQ ID NO: 63 or a variant thereof; and CDR-H3 having the sequence of SEQ ID NO: 64 or a variant thereof; and / or, a light chain variable region (VL) comprising the following three CDRs: CDR-L1 having the sequence of SEQ ID NO: 59 or a variant thereof; CDR-L2 having the sequence of SEQ ID NO: 60 or a variant thereof;A CDR-L3 comprising the sequence of SEQ ID NO: 61 or a variant thereof, wherein the variant described in any one of (1a), (1b-i), (1b-ii), (1c-i), (1c-ii), (1d), (2a), (2b), (2c-i), (2c-ii), (2d), (3a), (3b), (3c-i), (3c-ii), (3d), (4a), (4b), (4c-i), (4c-ii), and (4d) has a substitution, deletion, or addition of one or several amino acids (e.g., a substitution, deletion, or addition of 1, 2, or 3 amino acids) compared to the sequence from which the variant is derived; preferably, said substitution is a conservative substitution, an antibody or its antigen-binding fragment. Claim 3 In claim 1 or 2, the antibody or its antigen-binding fragment comprises: (ai) a VH comprising the sequence presented in SEQ ID NO: 54 or a variant thereof and / or a VL comprising the sequence presented in SEQ ID NO: 55 or a variant thereof; (a-ii) a VH comprising the sequence presented in SEQ ID NO: 52 or a variant thereof and / or a VL comprising the sequence presented in SEQ ID NO: 53 or a variant thereof; (bi) a VH comprising the sequence presented in SEQ ID NO: 3 or a variant thereof and / or a VL comprising the sequence presented in SEQ ID NO: 4 or a variant thereof; (b-ii) a VH comprising the sequence presented in SEQ ID NO: 1 or a variant thereof and / or a VL comprising the sequence presented in SEQ ID NO: 2 or a variant thereof; (ci) a VH comprising the sequence presented in SEQ ID NO: 9 or a variant thereof and / or a VL comprising the sequence presented in SEQ ID NO: 10 or a variant thereof; (c-ii) a VH presented in SEQ ID NO: 7 VH comprising a sequence or a variant thereof and / or VL comprising the sequence presented in SEQ ID NO: 8 or a variant thereof; (di) VH comprising the sequence presented in SEQ ID NO: 50 or a variant thereof and / or VL comprising the sequence presented in SEQ ID NO: 51 or a variant thereof; or (d-ii) comprising a VH containing the sequence presented in SEQ ID NO: 48 or a variant thereof and / or a VL containing the sequence presented in SEQ ID NO: 49 or a variant thereof, wherein the variant has a sequence identity of 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% with respect to the sequence from which the variant is derived, or has one or more amino acid substitutions, deletions, or additions (e.g., substitutions, deletions, or additions of 1, 2, 3, 4, or 5 amino acids) compared to the sequence from which the variant is derived;Preferably, the substitution is a conservative substitution of an antibody or its antigen-binding fragment. Claim 4 An antibody or its antigen-binding fragment according to any one of claims 1 to 3, wherein the antibody or its antigen-binding fragment is a murine antibody, a chimeric antibody, or a humanized antibody. Claim 5 In any one of claims 1 to 4, the antibody or its antigen-binding fragment further comprises a constant region of human immunoglobulin or a constant region derived from human immunoglobulin; preferably, the heavy chain of the antibody or its antigen-binding fragment comprises a heavy chain constant region of human immunoglobulin (e.g., IgG1, IgG2, IgG3, or IgG4) or a heavy chain constant region derived from human immunoglobulin; Preferably, the antibody or its antigen-binding fragment comprises a wild-type Fc region, or a mutated or chemically modified Fc region having an effector function altered compared to the wild-type Fc region; preferably, the antibody or its antigen-binding fragment comprises a variant of a human IgG1 heavy chain constant region, said variant having substitutions of Leu234Ala, Leu235Ala, and Gly237Ala (positions according to the EU numbering system) compared to the wild-type sequence from which said variant is derived; preferably, the light chain of the antibody or its antigen-binding fragment comprises a light chain constant region of human immunoglobulin, or a light chain constant region derived from human immunoglobulin (e.g., κ or λ); preferably, the antibody or its antigen-binding fragment comprises a heavy chain constant region (CH) presented in SEQ ID NO: 5 or its variant, said variant having a conservative substitution of up to 20 amino acids compared to SEQ ID NO: 5 (e.g., max Conservative substitutions of 15, at most 10, or at most 5 amino acids; e.g., conservative substitutions of 1, 2, 3, 4, or 5 amino acids; preferably, said antibody or its antigen-binding fragment comprises the light chain constant region (CL) presented in SEQ ID NO: 6 or its variant, said variant having conservative substitutions of up to 20 amino acids compared to SEQ ID NO: 6 (e.g., conservative substitutions of up to 15, at most 10, or at most 5 amino acids; e.g., conservative substitutions of 1, 2, 3, 4, or 5 amino acids);More preferably, the antibody or its antigen-binding fragment comprises the heavy chain constant region (CH) presented in SEQ ID NO: 5 and the light chain constant region (CL) presented in SEQ ID NO: 6.; Claim 6 In claim 5, the heavy chain constant region (CH) presented in SEQ ID NO: 5 or its variant is an antibody or its antigen-binding fragment lacking C-terminal lysine. Claim 7 In any one of claims 1 to 6, the antibody or its antigen-binding fragment comprises: (1) a heavy chain comprising the VH presented in SEQ ID NO: 54 and the heavy chain constant region (CH) presented in SEQ ID NO: 5; and a light chain comprising the VL presented in SEQ ID NO: 55 and the light chain constant region (CL) presented in SEQ ID NO: 6; (2) a heavy chain comprising the VH presented in SEQ ID NO: 52 and the heavy chain constant region (CH) presented in SEQ ID NO: 5; and a light chain comprising the VL presented in SEQ ID NO: 53 and the light chain constant region (CL) presented in SEQ ID NO: 6; (3) a heavy chain comprising the VH presented in SEQ ID NO: 3 and the heavy chain constant region (CH) presented in SEQ ID NO: 5; and a light chain comprising the VL presented in SEQ ID NO: 4 and the light chain invariant region (CL) presented in SEQ ID NO: 6; (4) a heavy chain comprising the VH presented in SEQ ID NO: 1 and the heavy chain invariant region (CH) presented in SEQ ID NO: 5; and a light chain comprising the VL presented in SEQ ID NO: 2 and the light chain invariant region (CL) presented in SEQ ID NO: 6; (5) a heavy chain comprising the VH presented in SEQ ID NO: 9 and the heavy chain invariant region (CH) presented in SEQ ID NO: 5; and a light chain comprising the VL presented in SEQ ID NO: 10 and the light chain invariant region (CL) presented in SEQ ID NO: 6; (6) a heavy chain comprising the VH presented in SEQ ID NO: 7 and the heavy chain invariant region (CH) presented in SEQ ID NO: 5; and a light chain comprising the VL presented in SEQ ID NO: 8 and the light chain invariant region (CL) presented in SEQ ID NO: 6; (7) a heavy chain comprising the VH presented in SEQ ID NO: 50 and the heavy chain invariant region (CH) presented in SEQ ID NO: 5; and a light chain comprising the VL presented in SEQ ID NO: 51 and the light chain invariant region (CL) presented in SEQ ID NO: 6; or (8) a heavy chain comprising the VH presented in SEQ ID NO: 48 and the heavy chain invariant region (CH) presented in SEQ ID NO: 5;An antibody or its antigen-binding fragment comprising a light chain comprising the VL presented in SEQ ID NO: 49 and the light chain constant region (CL) presented in SEQ ID NO: 6.; Claim 8 An antibody or its antigen-binding fragment, wherein the N-terminal glutamine of VH comprising the sequence presented in SEQ ID NO: 54, SEQ ID NO: 52, SEQ ID NO: 3, SEQ ID NO: 1, SEQ ID NO: 9, SEQ ID NO: 7, SEQ ID NO: 50, or SEQ ID NO: 48 or a variant thereof undergoes cyclization to form pyroglutamic acid or pyroglutamate. Claim 9 An antibody or its antigen-binding fragment according to any one of claims 1 to 8, wherein the antibody or its antigen-binding fragment comprises: (1) a heavy chain having the sequence presented in SEQ ID NO: 88 and a light chain having the sequence presented in SEQ ID NO: 89; (2) a heavy chain having the sequence presented in SEQ ID NO: 40 and a light chain having the sequence presented in SEQ ID NO: 41; (3) a heavy chain having the sequence presented in SEQ ID NO: 42 and a light chain having the sequence presented in SEQ ID NO: 43; or (4) a heavy chain having the sequence presented in SEQ ID NO: 86 and a light chain having the sequence presented in SEQ ID NO:

87. Claim 10 An antibody or its antigen-binding fragment, wherein the heavy chain N-terminal glutamine having the sequence presented in SEQ ID NO: 88, SEQ ID NO: 40, SEQ ID NO: 42 or SEQ ID NO: 86 or a variant thereof undergoes a cyclization reaction to form pyroglutamic acid or pyroglutamate. Claim 11 An antibody or its antigen-binding fragment according to any one of claims 1 to 10, wherein the antibody or its antigen-binding fragment is selected from ScFv, Fab, Fab', Fab'-SH, F(ab')2, Fv fragment, disulfide-linked Fv (dsFv), diabody, bispecific antibody, and multispecific antibody. Claim 12 In any one of claims 1 to 11, the antibody or its antigen-binding fragment has a label; preferably, the antibody or its antigen-binding fragment has a detectable label, e.g., an enzyme (e.g., horseradish peroxidase), a radionuclide, a fluorescent dye, a luminescent substance (e.g., a chemiluminescent substance), or biotin. Claim 13 In any one of claims 1 to 12, the antibody or its antigen-binding fragment has one or more features selected from the following: (1) less than about 100 ng / mL, e.g., less than about 80 ng / mL, less than 50 ng / mL, less than 20 ng / mL, less than 15 ng / mL, less than 14 ng / mL, less than 13 ng / mL, less than 12 ng / mL, less than 11 ng / mL, less than 10 ng / mL, less than 9 ng / mL, less than 8 ng / mL, less than 7 ng / mL, less than 6 ng / mL, less than 5 ng / mL, less than 4 ng / mL or less EC 50 , preferably, EC measured by ELISA 50 (1) binding to ADAM9 (e.g., human or monkey ADAM9); (2) binding to ADAM9 (e.g., human or monkey ADAM9) at a KD of less than about 100 nM, e.g., less than about 90 nM, less than 80 nM, less than 70 nM, less than 60 nM, less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 15 nM, less than 10 nM, less than 5 nM or less than the KD, preferably, as measured by BLI (bio-layer interferometry) (e.g., ForteBio Octet®); (3) having CDC activity, such as inducing apoptosis of ADAM9-expressing cells (e.g., tumor cells) via CDC; (4) not having ADCC activity; (5) induction of ADAM9 internalization, e.g., determined by flow cytometry; (6) ADAM12 and does not bind to ADAM15 protein; (7) inhibition of cell proliferation (e.g., tumor cells); and / or (8) inhibition of tumor growth. Claim 14 An isolated nucleic acid molecule comprising an antibody according to any one of claims 1 to 13 or its antigen-binding fragment, its heavy chain and / or light chain, or a nucleotide sequence encoding its heavy chain variable region and / or light chain variable region. Claim 15 Claim 14 comprises a nucleic acid molecule encoding a heavy chain variable region of the antibody and / or a nucleic acid molecule encoding a light chain variable region of the antibody, wherein (a) the nucleic acid molecule encoding the heavy chain variable region of the antibody comprises (i) a nucleotide sequence presented in SEQ ID NO: 92, (ii) a sequence substantially identical to SEQ ID NO: 92 (e.g., a sequence having at least about 85%, 90%, 95%, 99% or more sequence identity with respect to SEQ ID NO: 92, or a sequence having one or more nucleotide substitutions), or (iii) a degenerate sequence of (i) or (ii); and / or the nucleic acid molecule encoding the light chain variable region of the antibody comprises (iv) the nucleotide sequence presented in SEQ ID NO: 93, (v) a sequence substantially identical to SEQ ID NO: 93 (e.g., a sequence having at least about 85%, 90%, 95%, 99% or more sequence identity with SEQ ID NO: 93, or a sequence having one or more nucleotide substitutions), or (vi) the degenerate sequence of (iv) or (v); or (b) the nucleic acid molecule encoding the heavy chain variable region of the antibody comprises (i) the nucleotide sequence presented in SEQ ID NO: 44, (ii) a sequence substantially identical to SEQ ID NO: 44 (e.g., a sequence having at least about 85%, 90%, 95%, 99% or more sequence identity with SEQ ID NO: 44, or a sequence having one or more nucleotide substitutions), or (iii) the degenerate sequence of (i) or (ii);and / or the nucleic acid molecule encoding the light chain variable region of the antibody comprises (iv) the nucleotide sequence presented in SEQ ID NO: 45, (v) a sequence substantially identical to SEQ ID NO: 45 (e.g., a sequence having at least about 85%, 90%, 95%, 99% or more sequence identity with SEQ ID NO: 45, or a sequence having one or more nucleotide substitutions), or (vi) the degenerate sequence of (iv) or (v); or (c) the nucleic acid molecule encoding the heavy chain variable region of the antibody comprises (i) the nucleotide sequence presented in SEQ ID NO: 46, (ii) a sequence substantially identical to SEQ ID NO: 46 (e.g., a sequence having at least about 85%, 90%, 95%, 99% or more sequence identity with SEQ ID NO: 46, or a sequence having one or more nucleotide substitutions), or (iii) the degenerate sequence of (i) or (ii); and / or the nucleic acid molecule encoding the light chain variable region of the antibody comprises (iv) the nucleotide sequence presented in SEQ ID NO: 47, (v) a sequence substantially identical to SEQ ID NO: 47 (e.g., a sequence having at least about 85%, 90%, 95%, 99% or more sequence identity with SEQ ID NO: 47, or a sequence having one or more nucleotide substitutions), or (vi) the degenerate sequence of (iv) or (v); or (d) the nucleic acid molecule encoding the heavy chain variable region of the antibody comprises (i) the nucleotide sequence presented in SEQ ID NO: 90, (ii) a sequence substantially identical to SEQ ID NO: 90 (e.g., a sequence having at least about 85%, 90%, 95%, 99% or more sequence identity with SEQ ID NO: 90, or a sequence having one or more nucleotide substitutions), or (iii) the degenerate sequence of (i) or (ii);and / or the isolated nucleic acid molecule encoding the light chain variable region of the antibody comprises (iv) the nucleotide sequence presented in SEQ ID NO: 91, (v) a sequence substantially identical to SEQ ID NO: 91 (e.g., a sequence having at least about 85%, 90%, 95%, 99% or more sequence identity with respect to SEQ ID NO: 91, or a sequence having one or more nucleotide substitutions), or (vi) the degenerate sequence of (iv) or (v).; Claim 16 A vector comprising a nucleic acid molecule according to claim 14 or 15, wherein the vector is preferably a cloning vector or an expression vector. Claim 17 A host cell comprising a nucleic acid molecule according to claim 14 or 15 or a vector according to claim 16. Claim 18 A method for producing an antibody or its antigen-binding fragment according to any one of claims 1 to 13, comprising the steps of: culturing a host cell according to claim 17 under conditions allowing the expression of said antibody or its antigen-binding fragment; and recovering said antibody or its antigen-binding fragment from a culture of said cultured host cell. Claim 19 A conjugate comprising an antibody or its antigen-binding fragment according to any one of claims 1 to 13 and a conjugate moiety connected thereto, wherein, preferably, the conjugate moiety is selected from a detectable label (e.g., a radioisotope, a fluorescent substance, a luminescent substance, a colored substance, or an enzyme) or a therapeutic agent (e.g., a cytotoxic agent, a cytokine, a toxin, or a radionuclide). Claim 20 A multispecific antibody comprising an antibody according to any one of claims 1 to 13 or an antigen-binding fragment thereof, wherein, preferably, the multispecific antibody comprises an antibody according to any one of claims 1 to 13 or an antigen-binding fragment thereof as a first antigen-binding domain and further comprises at least one second antigen-binding domain for another target; preferably, the multispecific antibody is a bispecific antibody, a trispecific antibody, or a quadrspecific antibody. Claim 21 A chimeric antigen receptor comprising an antibody or its antigen-binding fragment (e.g., ScFv) according to any one of claims 1 to 13, a transmembrane domain, and one or more intracellular T cell signaling domains. Claim 22 A pharmaceutical composition comprising an antibody or its antigen-binding fragment according to any one of claims 1 to 13, or an isolated nucleic acid molecule according to claim 14 or 15, or a vector according to claim 16, or a host cell according to claim 17, or a conjugate according to claim 19, or a multispecific antibody according to claim 20, or a chimeric antigen receptor according to claim 21 or a host cell expressing said chimeric antigen receptor, and a pharmaceutically acceptable carrier and / or excipient, wherein, preferably, the pharmaceutical composition further comprises an additional pharmaceutical active agent; preferably, said additional pharmaceutical active agent is a drug having anti-tumor activity; preferably, said additional pharmaceutical active agent is from an ADAM9 inhibitor, an EGFR inhibitor, a HER2 inhibitor, a HER3 inhibitor, a HER4 inhibitor, an IGFR-1 inhibitor, an mTOR inhibitor, a PI3 kinase inhibitor, a c-met inhibitor or a VEGF inhibitor, a chemotherapeutic agent, or any combination thereof. A pharmaceutical composition selected; preferably, said antibody or its antigen-binding fragment and additional pharmaceutical active agent provided as separate components or as mixed components. Claim 23 A diagnostic or therapeutic kit comprising an antibody or its antigen-binding fragment according to any one of claims 1 to 13, or an isolated nucleic acid molecule according to claim 14 or 15, or a vector according to claim 16, or a host cell according to claim 17, or a conjugate according to claim 19, or a multispecific antibody according to claim 20, or a chimeric antigen receptor according to claim 21 or a host cell expressing said chimeric antigen receptor, or a pharmaceutical composition according to claim 22, and optionally an instruction manual and / or a delivery device. Claim 24 In the manufacture of a drug for inhibiting the proliferation of cells (e.g., ADAM9-expressing cells, e.g., tumor cells) or for preventing and / or treating and / or providing adjuvant treatment for tumors, neurodegenerative diseases, retinal diseases or inflammation, wherein, for the use of an antibody or its antigen-binding fragment according to any one of claims 1 to 13, or an isolated nucleic acid molecule according to claim 14 or 15, or a vector according to claim 16, or a host cell according to claim 17, or a conjugate according to claim 19, or a multispecific antibody according to claim 20, or a chimeric antigen receptor or a host cell expressing said chimeric antigen receptor according to claim 21, or a pharmaceutical composition according to claim 22, preferably, said antibody or its antigen-binding fragment, isolated nucleic acid molecule, vector, host cell, conjugate, multispecific antibody, or pharmaceutical composition is administered, for example, simultaneously, individually, or sequentially in combination with an additional pharmaceutical active agent; preferably, The additional pharmaceutical active agent is a drug having anti-tumor activity; preferably, the additional pharmaceutical active agent is selected from ADAM9 inhibitors, EGFR inhibitors, HER2 inhibitors, HER3 inhibitors, HER4 inhibitors, IGFR-1 inhibitors, mTOR inhibitors, PI3 kinase inhibitors, c-met inhibitors, or VEGF inhibitors, chemotherapy agents, or any combination thereof. Claim 25 In claim 24, the tumor is an ADAM9-positive tumor; preferably, the tumor is selected from glioma, cervical cancer, oral squamous cell carcinoma, liver cancer, melanoma, prostate cancer, pancreatic ductal adenocarcinoma, gastric cancer, lung cancer such as non-small cell lung cancer, pancreatic cancer, ovarian cancer, colorectal cancer, head and neck cancer, breast cancer such as triple-negative breast cancer, or any combination thereof. Claim 26 A method for inhibiting cell proliferation comprising the step of contacting a cell with an antibody or its antigen-binding fragment according to any one of claims 1 to 13, or an isolated nucleic acid molecule according to claim 14 or 15, or a vector according to claim 16, or a host cell according to claim 17, or a conjugate according to claim 19, or a multispecific antibody according to claim 20, or a chimeric antigen receptor according to claim 21 or a host cell expressing said chimeric antigen receptor, or a pharmaceutical composition according to claim 22, wherein, preferably, the cell is an ADAM9-expressing cell, e.g., a tumor cell; and preferably, the tumor cell overexpresses ADAM9. Claim 27 A method for preventing and / or treating and / or adjuvantly treating a tumor, neurodegenerative disease, retinal disease, or inflammation in a subject, comprising the step of administering an effective amount of an antibody or its antigen-binding fragment according to any one of claims 1 to 13, or an isolated nucleic acid molecule according to claim 14 or 15, or a vector according to claim 16, or a host cell according to claim 17, or a conjugate according to claim 19, or a multispecific antibody according to claim 20, or a chimeric antigen receptor according to claim 21 or a host cell expressing said chimeric antigen receptor, or a pharmaceutical composition according to claim 22, to a subject requiring such treatment. Claim 28 The method of claim 27 further comprises the step of applying a second therapy selected from surgery, chemotherapy, radiation therapy, immunotherapy, gene therapy, DNA therapy, RNA therapy, nanotherapy, viral therapy, adjuvant therapy, and any combination thereof to the subject, wherein optionally, the second therapy may be applied individually or sequentially, simultaneously with the method according to claim 27. Claim 29 A method according to claim 27 or 28, wherein the tumor is an ADAM9-positive tumor; preferably, the tumor is selected from glioma, cervical cancer, oral squamous cell carcinoma, liver cancer, melanoma, prostate cancer, pancreatic ductal adenocarcinoma, gastric cancer, lung cancer such as non-small cell lung cancer, pancreatic cancer, ovarian cancer, colorectal cancer, head and neck cancer, breast cancer such as triple-negative breast cancer, or any combination thereof. Claim 30 A method for detecting the presence or level of ADAM9 in a sample comprises the steps of contacting said sample with said antibody or said antigen-binding fragment under conditions allowing the formation of a complex between said antibody or said antigen-binding fragment and ADAM9 according to any one of claims 1 to 13, and detecting said formation of said complex; preferably, said method is used to diagnose a tumor, e.g., ADAM9-positive tumor, e.g., glioma, cervical cancer, oral squamous cell carcinoma, liver cancer, melanoma, prostate cancer, pancreatic ductal adenocarcinoma, gastric cancer, lung cancer, e.g., non-small cell lung cancer, pancreatic cancer, ovarian cancer, colorectal cancer, head and neck cancer, breast cancer, e.g., triple-negative breast cancer, or any combination thereof; preferably, said method comprises the steps of detecting the expression level of ADAM9 in a test sample derived from a subject, and comparing said expression level with a reference value, wherein an increase in said expression level compared with said reference value indicates a tumor. Claim 31 In the manufacture of a detection kit for detecting the presence or level of ADAM9 in a sample and / or diagnosing a tumor, the use of an antibody or its antigen-binding fragment according to any one of claims 1 to 13, or an isolated nucleic acid molecule according to claim 14 or 15, or a vector according to claim 16, or a host cell according to claim 17, or a conjugate according to claim 19, or a multispecific antibody according to claim 20, preferably, said tumor is an ADAM9-positive tumor; preferably, said tumor is selected from glioma, cervical cancer, oral squamous cell carcinoma, liver cancer, melanoma, prostate cancer, pancreatic ductal adenocarcinoma, gastric cancer, lung cancer such as non-small cell lung cancer, pancreatic cancer, ovarian cancer, colorectal cancer, head and neck cancer, breast cancer such as triple-negative breast cancer, or any combination thereof.

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