Anti-tissue factor protein antibody and antibody-drug conjugate thereof

Abstract

Provided are an anti-TF antibody or an antigen-binding fragment thereof, an antibody-drug conjugate containing the antibody or antigen-binding fragment thereof, a pharmaceutical composition containing the antibody-drug conjugate, and a method for preparing the antibody-drug conjugate and the use thereof (e.g., the use in the treatment and / or prevention of TF-positive cancers).

Description

An antibody against tissue factor protein and antibody-drug conjugate thereof

[0001] Reference to Related Applications

[0002] This application claims priority to Chinese Patent Application No. 202411379243.0, filed September 30, 2024, the entire contents of which are incorporated by reference herein in its entirety and for all purposes. TECHNICAL FIELD

[0003] The present application relates to the field of biological medicine, in particular to an antibody against tissue factor protein or an antigen-binding fragment thereof, an antibody-drug conjugate comprising the antibody or the antigen-binding fragment thereof, a pharmaceutical composition comprising the antibody-drug conjugate, and a preparation method and uses (e.g., uses in treating and / or preventing TF-positive cancer) of the antibody-drug conjugate. BACKGROUND

[0004] Tissue factor (TF, herein abbreviated as "TF", also known as FIII or CD142 in the art) is a 47 kDa membrane-bound glycoprotein called thromboplastin, expressed on the cell membrane of smooth muscle cells and fibroblasts of the outer membrane of blood vessels, not expressed in endothelial cells of the inner wall of normal tissue blood vessels. TF binds to FVII (coagulation factor VII) to activate FVII to FVIIa, and the TF-FVIIa complex converts FX (coagulation factor X) to FXa, thereby activating the downstream signaling pathway of TF and playing a role in blood coagulation.

[0005] TF is limitedly expressed in normal tissues, but highly expressed in tumor tissues, such as cervical cancer, pancreatic cancer, head and neck cancer, endometrial cancer, esophageal cancer, prostate cancer, non-small cell lung cancer, and ovarian cancer, so that drugs targeting TF can be delivered to tumor cells, and can be an ideal target for antibody-drug conjugate (ADC) drugs. The ADC targeting TF can selectively kill tumor cells expressing TF and inhibit tumor growth in vivo. Tivdak (Tisotumab Vedotin) is an ADC anti-cancer drug targeting TF, which aims to target the TF antigen on cancer cells and deliver the cytotoxic agent MMAE (monomethyl auristatin E) directly into cancer cells. In 2021, the U.S. Food and Drug Administration accelerated the approval of the drug for patients with recurrent or metastatic cervical cancer who have received chemotherapy or have disease progression after chemotherapy. However, in clinical trials, 60% of cervical cancer patients treated with Tivdak had ocular adverse reactions, 62% of patients had bleeding events, 42% of patients had peripheral neuropathy, and 8% of patients had grade 3 peripheral neuropathy, indicating that the side effects of the drug were significant.

[0006] Therefore, there is still a need to develop safer and more effective ADC drugs to reduce the adverse reactions such as bleeding and peripheral neuropathy in patients to some extent, and to provide safer and more effective drug options for cancer patients.

[0007] SUMMARY

[0008] In a first aspect, the present application provides an anti-tissue factor (TF) antibody or an antigen binding fragment thereof, the antibody or antigen binding fragment comprising a heavy chain variable region comprising three complementarity determining regions HCDR1, HCDR2 and HCDR3, and a light chain variable region comprising three complementarity determining regions LCDR1, LCDR2 and LCDR3, wherein:

[0009] (1) the amino acid sequence of HCDR1 is SEQ ID NO. 5 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO. 5, the amino acid sequence of HCDR2 is SEQ ID NO. 6 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO. 6, the amino acid sequence of HCDR3 is SEQ ID NO. 7 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO. 7, the amino acid sequence of LCDR1 is SEQ ID NO. 14 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO. 14, the amino acid sequence of LCDR2 is SEQ ID NO. 15 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO. 15, and the amino acid sequence of LCDR3 is SEQ ID NO. 16 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO. 16; or

[0010] (2) the amino acid sequence of HCDR1 is set forth in SEQ ID NO. 20 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 20, the amino acid sequence of HCDR2 is set forth in SEQ ID NO. 21 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 21, the amino acid sequence of HCDR3 is set forth in SEQ ID NO. 22 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 22, the amino acid sequence of LCDR1 is set forth in SEQ ID NO. 27 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 27, the amino acid sequence of LCDR2 is set forth in SEQ ID NO. 28 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 28, and the amino acid sequence of LCDR3 is set forth in SEQ ID NO. 29 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 29;

[0011] wherein the HCDR and LCDR sequences are according to the Kabat definition.

[0012] In a second aspect, the present application provides an isolated nucleic acid molecule encoding the anti-TF antibody or antigen binding fragment thereof according to the first aspect.

[0013] In a third aspect, the present application provides a vector (e.g., an expression vector) comprising the nucleic acid molecule of the second aspect.

[0014] In a fourth aspect, the present application provides a cell comprising the nucleic acid molecule of the second aspect or the vector of the third aspect.

[0015] In a fifth aspect, the present application provides an antibody-drug conjugate having the structure of Formula (I), or stereoisomers, tautomers, and pharmaceutically acceptable salts thereof:

[0016] A-(L-D)d (I)

[0017] wherein:

[0018] A is the anti-TF antibody or antigen binding fragment thereof according to the first aspect;

[0019] D is a pharmaceutically active molecule, preferably a molecule having a therapeutic activity for a disease, more preferably a molecule having an anticancer activity, most preferably a small molecule having an anticancer activity;

[0020] L is a linker moiety, one end of which is connected to A, and one end of which is connected to the pharmaceutically active molecule D;

[0021] d is selected from an integer (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) or a decimal number from 1 to 12.

[0022] In a sixth aspect, the present application provides a pharmaceutical composition comprising the anti-tissue factor (TF) antibody or antigen-binding fragment thereof of the first aspect, or the antibody-drug conjugate or the pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled of the fifth aspect, or the pharmaceutical composition of the sixth aspect, and a pharmaceutically acceptable carrier or excipient.

[0023] In a seventh aspect, the present application provides use of the anti-tissue factor (TF) antibody or antigen-binding fragment thereof of the first aspect, or the antibody-drug conjugate or the pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled of the fifth aspect, or the pharmaceutical composition of the sixth aspect, in the manufacture of a medicament for treating and / or preventing a TF-mediated disease or disorder.

[0024] In an eighth aspect, the present application provides a method of treating and / or preventing a TF-mediated disease or disorder, comprising administering to a subject in need thereof an effective amount of the anti-tissue factor (TF) antibody or antigen-binding fragment thereof of the first aspect, or the antibody-drug conjugate or the pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled of the fifth aspect, or the pharmaceutical composition of the sixth aspect.

[0025] In a ninth aspect, the present application provides a pharmaceutical combination comprising the anti-tissue factor (TF) antibody or antigen-binding fragment thereof of the first aspect, or the antibody-drug conjugate or the pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled of the fifth aspect, or the pharmaceutical composition of the sixth aspect, and one or more additional therapeutic agents.

[0026] In a tenth aspect, the present application provides a kit comprising the anti-tissue factor (TF) antibody or antigen-binding fragment thereof of the first aspect, or the antibody-drug conjugate or the pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled of the fifth aspect, or the pharmaceutical composition of the sixth aspect. BRIEF DESCRIPTION OF DRAWINGS

[0027] Figure 1 shows curves of the effect of exemplary antibodies and reference antibodies of the present application on FXa generation.

[0028] Figure 2 shows the effect of exemplary antibodies and reference antibodies of the present application on coagulation activity.

[0029] Figure 3 shows endocytosis activity of exemplary antibodies and reference antibodies of the present application.

[0030] Figures 4A and 4B show single crystal diffraction patterns of compound 12.

[0031] Figure 5 shows inhibition curves of exemplary ADCs and reference ADCs of the present application on human pancreatic cancer HPAF-II mouse xenograft tumor volume.

[0032] Figure 6 shows inhibition curves of exemplary ADCs and reference ADCs of the present application on human pharyngeal squamous carcinoma FADU mouse xenograft tumor volume.

[0033] Figure 7 shows inhibition curves of exemplary ADCs of the present application on human cervical carcinoma CaSki mouse xenograft tumor volume.

[0034] Figure 8 shows inhibition curves of exemplary ADCs and reference ADCs of the present application on human pharyngeal squamous carcinoma HT-3 mouse xenograft tumor volume.

[0035] DETAILED DESCRIPTION

[0036] TERMS

[0037] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the application belongs. The abbreviations for amino acid residues are those standard three letter and / or one letter codes used in the art to designate one of the 20 common L-amino acids.

[0038] Notwithstanding that the numerical ranges and parameters setting forth the broadest scope of the application are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. Any numerical value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements. Moreover, all ranges disclosed herein are to be understood to encompass any and all subranges subsumed therein. For example, a range of "1 to 10" is to be understood to include any and all subranges between (and including) the minimum value of 1 and the maximum value of 10, that is, any and all subranges beginning with a minimum of 1 or more, e.g. 1 to 6.1, and ending with a maximum of 10 or less, e.g. 5.5 to 10. Additionally, any reference to a reference cited herein is to be understood as an incorporation by reference in its entirety.

[0039] The term "subject" or "individual" as used herein refers to a mammal, such as a human, but can also be another animal, such as a wild animal, a domestic animal or a laboratory animal (e.g., a chimpanzee, a monkey, a rat, a mouse, a rabbit, a guinea pig, a woodchuck, a ground squirrel, etc.).

[0040] The term "antigen" as used herein is a predetermined target to which an antibody can selectively bind. Examples of antigens include, but are not limited to, a polypeptide, a saccharide, a nucleic acid, a lipid, a hapten, or other naturally occurring or synthetic compound.

[0041] Broadly speaking, an "antibody" can refer to an immunoglobulin molecule capable of specific binding to a target, via at least one antigen recognition site located in the variable region of the immunoglobulin molecule, and thus encompasses both intact antibodies / full length antibodies, antibody single chains, or any antigen binding fragment of an antibody (also known as "antigen binding portion"). When "antibody" and "antigen binding fragment / antigen binding portion" appear in the same context, "antibody" can be understood as the complete entity in relation to "antigen binding fragment / antigen binding portion", both together corresponding to the broad concept of antibody.

[0042] A "full-length antibody" is a protein comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain comprises a heavy chain variable region (abbreviated as VH) and a heavy chain constant region (abbreviated as CH). The heavy chain constant region comprises three domains, CH1, CH2 and CH3. Each light chain comprises a light chain variable region (abbreviated as VL) and a light chain constant region (abbreviated as CL). The light chain constant region comprises one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with regions that are more conserved, termed framework regions (FR). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. These variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The constant regions of the antibodies can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system. A full-length antibody can be of any class, e.g., IgD, IgE, IgG, IgA or IgM (or a subclass thereof), although the antibody need not be of any particular class. The assignment of an immunoglobulin to a particular class can be determined based on the amino acid sequence of the constant domain of the heavy chain. Generally, immunoglobulins of five major classes, IgA, IgD, IgE, IgG, and IgM, and several of these can be further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains that correspond to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known. Chimeric or humanized antibodies are also encompassed within the antibodies according to the present application. As is well known in the art, the complementarity determining regions (CDRs, typically CDR1, CDR2 and CDR3) are the regions of the variable region that most significantly affect the affinity and specificity of an antibody. There are several common definitions of the CDR amino acid sequences of a VH or VL, such as the Kabat definition, the IMGT definition, the Chothia definition, etc. For a given variable region amino acid sequence of an antibody, the CDR amino acid sequences can be determined according to different definitions. In embodiments of the present application, the CDR amino acid sequences are defined according to the Kabat definition. For a given variable region amino acid sequence of an antibody, the CDR amino acid sequences can be analyzed in several ways.

[0043] The term "murine antibody" refers to an antibody derived from the fusion of B cells from immunized mice with myeloma cells, the screening of murine hybrid fusion cells that can both proliferate indefinitely and secrete antibodies, and the subsequent selection, production and purification of the antibodies obtained. Murine antibodies are generally immunogenic, and thus, if subsequent administration to humans is desired, they are typically subjected to humanization.

[0044] The term "humanized antibody" means an antibody obtained by grafting CDR sequences derived from another mammalian species, such as the mouse germline, onto human framework sequences. In order to preserve binding affinity, some residues of the framework (called FR) segments can be modified. Humanized antibodies or fragments thereof according to the application can be prepared by techniques known to those skilled in the art.

[0045] The term "chimeric antibody" refers to an antibody in which the variable region sequences are of one species and the constant region sequences are of another species, for example, an antibody in which the variable region sequences are of a mouse antibody and the constant region sequences are of a human antibody. Chimeric antibodies according to the application or fragments thereof can be prepared by using genetic recombination techniques. For example, the chimeric antibody can be produced by cloning recombinant DNA comprising a promoter and a sequence encoding the variable region of a non-human, in particular mouse, monoclonal antibody according to the application, and a sequence encoding the constant region of a human antibody. The chimeric antibody according to the application encoded by this recombinant gene will be, for example, a murine-human chimera, the specificity of which is determined by the variable region derived from the murine DNA, and the isotype of which is determined by the constant region derived from the human DNA.

[0046] The term "monoclonal antibody" refers to an antibody obtained from a population of essentially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that can be present in minor amounts.

[0047] The terms "antigen binding fragment" or "antigen binding portion" or "antigen binding region" as used herein are used interchangeably and refer to a portion of an antibody comprising amino acid residues that interact with an antigen and confer to it the binding specificity and affinity for a particular antigen, and in particular to antibody fragments such as Fv, Fab, F(ab')2 or Fab', or any fragment that should be able to increase the half-life by chemical modification or by incorporation into liposomes, such as poly(alkylene)glycols such as polyethylene glycol ("PEGylation") (PEGylated fragments known as Fv-PEG, scFv-PEG, Fab-PEG, F(ab')2-PEG or Fab'-PEG) ("PEG" is polyethylene glycol) that have TF binding activity. Preferably, the antigen binding fragment will be composed of or will comprise part of the sequence of the heavy or light chain variable chain of the antibody from which it is derived, said part of the sequence being sufficient to retain the same binding specificity and sufficient affinity as the antibody from which it is derived, such an antigen binding fragment will comprise a minimum of 5 amino acids, preferably 10, 15, 25, 50 and 100 contiguous amino acids of the sequence of the antibody from which it is derived. Examples of antigen binding fragments include, but are not limited to: (1) Fab fragments, which can be monovalent fragments having a VL-CL chain and a VH-CH1 chain; (2) F(ab')2 fragments, which can be bivalent fragments having two Fab' fragments joined by a disulfide bridge in the hinge region (i.e., a dimer of Fab' fragments); (3) Fv fragments having a single arm of the VL and VH domains of an antibody; (4) VHH fragments consisting of a VH domain.

[0048] The term "single chain antibody (scFv) refers to a single polypeptide chain connected by a peptide linker between the VH and VL domains. (scFv)2 comprises two VH domains and two VL domains connected by a peptide linker, the two VL domains being combined with the two VH domains via disulfide bridges.

[0049] The term "Fc fragment", "Fc region", "Fc domain", "Fc portion" or similar terms refer to a portion of the constant region of the heavy chain of an antibody, including the hinge, the CH2 fragment and the CH3 fragment of the constant region.

[0050] The term "specifically binds" as used herein refers to a non-random binding reaction between two molecules, for example the binding of an antibody to an epitope of an antigen.

[0051] Generally, to prepare monoclonal antibodies or functional fragments thereof, especially of murine origin, reference can be made to the techniques described in particular in the manual "Antibodies" (Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor NY, pp. 726, 1988) or to the techniques described by Kohler and Milstein for the preparation from hybridoma cells (Nature, 256: 495-497, 1975).

[0052] In describing some embodiments involving mutation of an amino acid or nucleic acid sequence, the application uses the designation "XaaaY" (e.g., L234A, etc.), where "aaa" represents the sequence position of the amino acid or base (where a particular reference sequence is present, "aaa" represents the sequential position of the residue in the reference sequence; alternatively, the position numbering can be according to the art-accepted position numbering system, such as the EU numbering system, etc.), "X" represents the original amino acid or base at "aaa", and "Y" represents the changed amino acid or base at "aaa". As an example, when describing a L234A mutation in a human heavy chain constant region, this means that the leucine (L) at position 234, according to the EU numbering system for the human heavy chain constant region, is mutated to an alanine (A).

[0053] The term "isolated" biological component (e.g., nucleic acid, protein (including antibodies), or organelle) has been substantially separated or purified away from other biological components of the environment in which the component naturally occurs, e.g., the cell (i.e., other chromosomal and extra-chromosomal DNA and RNA, proteins, and organelles). Nucleic acids and proteins that have been "isolated" include nucleic acids and proteins purified by standard purification methods. The term also embraces nucleic acids and proteins prepared by recombinant expression in a host cell as well as chemically synthesized nucleic acids.

[0054] An "expression vector" is a vector that includes one or more expression control sequences, and an "expression control sequence" is a DNA sequence that controls and regulates transcription and / or translation of another DNA sequence.

[0055] The term "pharmaceutical composition" as used herein means a combination of at least one pharmaceutical together with optionally a pharmaceutically acceptable carrier or excipient that are combined in order to facilitate administration of the pharmaceutical. In certain embodiments, the pharmaceutical composition comprises combinations that are separated in time and / or space, as long as they are able to act together to achieve the purposes of the present application. For example, the components contained in the pharmaceutical composition (e.g., antibodies, nucleic acid molecules, combinations of nucleic acid molecules, and / or conjugates according to the present application) can be administered to an individual as a whole or separately. When the components contained in the pharmaceutical composition are administered separately to an individual, the components can be administered to the individual simultaneously or sequentially. For example, the pharmaceutically acceptable carrier is water, a buffered aqueous solution, an isotonic salt solution such as PBS (phosphate buffered saline), glucose, mannitol, dextrose, lactose, starch, magnesium stearate, cellulose, magnesium carbonate, 0.3% glycerol, hyaluronic acid, ethanol, or a polyalkylene glycol such as polypropylene glycol, a triglyceride, or the like. The type of pharmaceutically acceptable carrier used depends, inter alia, on whether the composition according to the present application is formulated for oral, nasal, intradermal, subcutaneous, intramuscular, or intravenous administration. The composition according to the present application can contain a wetting agent, emulsifying agent, or a buffer substance as an additive. The pharmaceutical composition or pharmaceutical preparation according to the present application can be administered by any suitable route, for example, orally, nasally, intradermally, subcutaneously, intramuscularly, or intravenously.

[0056] The term "effective amount" in the present application means an amount that brings about a therapeutic effect on a subject, for example, alleviation, reduction, or elimination of the symptoms or state of a disease, or delay or inhibition of the development of the symptoms or state of a disease in a subject to whom the amount is administered, as compared to a subject to whom the amount is not administered.

[0057] EC 50 The term "EC50" refers to the concentration of a drug, antibody, or toxin, etc. that is required to achieve 50% of the maximum biological effect after a specified exposure time. In pharmacology, it is used to characterize the activation of an agonist in vitro, and to indicate the blood concentration required to achieve half of the maximum biological effect in vivo. In some literature, EC 50 is also used to characterize the potency of a compound at the cellular level (both agonism and antagonism) and can be determined by methods such as ELISA. 50

[0058] The term "identity" or "homology" with respect to an amino acid or nucleic acid sequence is defined as the percentage of residues in an amino acid or nucleotide sequence variant that are identical with the reference sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent identity. Methods and computer programs for alignment are well known in the art.

[0059] ​As used herein, the term "cancer" or "cancerous" refers to a proliferative disorder disease caused by or characterized by cell proliferation that has lost susceptibility to normal growth controls. The term "cancer" includes neoplasms and any other proliferative disorder. Cancers of the same tissue type arise in the same tissue and can be classified into different subtypes based on their biological characteristics.

[0060] The term "antibody-drug conjugate" or "antibody conjugated drug" (ADC) refers to a compound in which a targeting ligand such as an antibody (e.g., a monoclonal antibody), an antibody fragment is linked to a biologically active molecule having a biological activity through a stable chemical linker compound.

[0061] The term "linker-drug compound" or "linker-drug compound" (L-D) refers to a partial structure consisting of a linker compound and a biologically active compound in an "antibody-drug conjugate".

[0062] The term "DAR value" or "DAR" or "d" as used herein refers to the ratio of a drug (D) and an antibody (A) in an antibody-drug conjugate, or the ratio of a linker-drug compound (L-D) and an antibody (A). When the DAR value or d is 8, it means that one molecule of an antibody is conjugated to 8 linker-drug compounds. When the DAR value or d is a decimal number, it means that the average number of linker-drug compounds (L-D) conjugated to an antibody in an antibody-drug conjugate.

[0063] The term "linker" refers to a fragment of a chemical structure that is connected to an antibody at one end and to a cytotoxic drug at the other end, which is represented by L, which is formed by a "linker compound" connecting an antibody and a biologically active compound, respectively. Exemplary linkers in the present application include a linking portion to an antibody, a spacer, a polypeptide molecule, and a self-cleaving fragment.

[0064] In some embodiments of the present application, the linkage of the linker-drug compound to the antibody can be performed by conventional coupling methods in the art, including: lysine coupling, inter-heavy chain disulfide coupling and directed coupling (Beck A, Reichert JM. Antibody-drug conjugates: Present and future; MAbs, 2014, 6: 15-17; McCombs J R, Owen S C. Antibody drug conjugates: design and selection of linker, payload and conjugation chemistry. The AAPS journal, 2015, 17: 339-351). For example, the linkage can be performed by inter-heavy chain disulfide coupling, i.e. the reaction of the thiol group (sulfur atom of cysteine residue) formed after reduction of one or more of the inter-heavy chain disulfide bond sites (two sites between heavy chains, two sites between heavy and light chains).

[0065] In some embodiments of the present application, the linkage of the linker L to the antibody is represented by L1, which is formed by the reaction of the L1’ group in the linker compound with the antibody, for example Or wherein * represents the linkage to the thiol group of the antibody, and ** represents the linkage to the spacer.

[0066] In the context of linker L, the terms “spacer” and “extension unit” are used interchangeably to mean the linking group between the antibody-binding moiety and the peptide residue moiety, which can be any divalent organic group, such as a chemical bond, C 1-10 alkylene, C 2-10 alkenylene, C 2-10 alkynylene, C 3-10 cycloalkyl, C 6-12 aryl, 5-12 membered heteroaryl, 5-12 membered heterocyclyl, or a combination of two or more of these groups; said C 1-10 alkylene, C 2-10 alkenylene, C 2-10 alkynylene, C 3-10 cycloalkyl, C 6-12 aryl, 5-12 membered heteroaryl, 5-12 membered heterocyclyl, or a combination of two or more of these groups and optionally interrupted by a carbonyl, O, S, N atom; said C 1-10 alkylene, C 2-10 alkenylene, C 2-10 alkynylene, C 3-10 cycloalkyl, C 6-12 aryl, 5-12 membered heteroaryl, 5-12 membered heterocyclyl can optionally be substituted by C1-6 alkyl, C 3-6 cycloalkyl, halogen atom, haloC 1-6 alkyl substituted. In some embodiments of the application, the spacer is represented by L2, the definition of L2 also applies to the spacer definition.

[0067] In the context of linker L, the term "peptide residue" can be a divalent peptide radical comprising 2 to 8 optionally substituted natural or non-natural, L-form or D-form amino acid residues, each of said amino acid residues can be the same or different, independently of each other selected from the group consisting of residues of alanine (Ala), cysteine (Cys), aspartic acid (Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly), histidine (His), isoleucine (lie), lysine (Lys), leucine (Leu), methionine (Met), asparagine (Asn), proline (Pro), glutamine (Gin), arginine (Arg), serine (Ser), threonine (Thr), valine (Val), tryptophan (Trp), tyrosine (Tyr), citrulline (Cit), valine (Nva), norleucine (Nle) or an analogue of the above amino acids. In some embodiments of the application, the peptide residue is represented by L3, the definition of L3 also applies to the peptide residue definition.

[0068] In the context of linker L, the term "self-cleaving fragment" can be understood in the meaning known in the art, such as the p-aminobenzyl alcohol carbonate fragment as conventionally used in the art, in the specific embodiments of the application, the "self-cleaving fragment" of the linkers of the application comprises a modification of the "hydrophilic fragment". In some embodiments of the application, the self-cleaving fragment is represented by L4, the definition of L4 also applies to the self-cleaving fragment definition.

[0069] The term "heteroatom" means nitrogen, oxygen, sulfur, halogen atom.

[0070] The term "halogen" or "halo" means F, CI, Br, I.

[0071] The term "a plurality of" in the term "one or more" includes two or more (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, etc.).

[0072] The term "C m -C n " means that the moiety has an integer number of carbon atoms in the range of m to n. For example "C1-C3" means that the group can have 1 carbon atom, 2 carbon atoms, or 3 carbon atoms.

[0073] The term "aryl" denotes monocyclic, bicyclic and tricyclic aromatic carbocyclic ring systems containing 6-16 carbon atoms, or 6-14 carbon atoms, or 6-12 carbon atoms, or 6-10 carbon atoms, preferably 6-10 carbon atoms. Examples of aryl groups can include, but are not limited to, phenyl, naphthyl, anthryl, phenanthryl or pyrenyl, and the like.

[0074] The term "heteroaryl" denotes an aromatic monocyclic or polycyclic ring system containing a 5-14 membered structure, or preferably a 5-10 membered structure, or preferably a 5-8 membered structure, more preferably a 5-6 membered structure, wherein 1, 2, 3 or more ring atoms are heteroatoms and the remaining atoms are carbon, the heteroatoms being independently selected from O, N or S, the number of heteroatoms preferably being 1, 2 or 3. Examples of heteroaryl groups include, but are not limited to, furanyl, thienyl, oxazolyl, thiazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, thiadiazolyl, triazinyl, phtalazinyl, quinolinyl, isoquinolinyl, pteridinyl, purinyl, indolyl, isoindolyl, indazolyl, benzofuranyl, benzothienyl, benzopyridyl, benzopyrimidyl, benzopyrazinyl, benzimidazolyl, benzophthalazinyl, pyrrolo[2,3-b]pyridyl, imidazo[l,2-a]pyridyl, pyrazolo[l,5-a]pyridyl, pyrazolo[l,5-a]pyrimidyl, imidazo[l,2-b]pyridazinyl, [l,2,4]triazolo[4,3-b]pyridazinyl, [l,2,4]triazolo[l,5-a]pyrimidyl, [l,2,4]triazolo[l,5-a]pyridyl, and the like.

[0075] The term "heterocyclyl" refers to a non-aromatic ring that is fully saturated or partially unsaturated (not fully unsaturated heteroaromatic) and can exist as a monocyclic, bridged, or spirocyclic ring. Unless otherwise indicated, the heterocycle is typically a 3- to 7-membered ring containing 1 to 3 heteroatoms independently selected from sulfur, oxygen, and / or nitrogen (preferably 1 or 2 heteroatoms, but not including -0-0-, -0-S-, or -S-S- moieties). Non-limiting examples of heterocyclyl groups include, but are not limited to, oxiranyl, tetrahydrofuranyl, dihydrofuranyl, pyrrolidinyl, N-methylpyrrolidinyl, dihydropyrrolyl, piperidinyl, piperazinyl, pyrazolidinyl, 4H-pyranyl, morpholinyl, thiomorpholinyl, tetrahydrothienyl, and the like.

[0076] The term "cycloalkyl" refers to a carbocyclic ring that is fully saturated and can exist as a monocyclic, bridged, or spirocyclic ring. Preferably contains 3-12 carbon atoms (i.e. C 3-12 cycloalkyl), more preferably contains 3-10 carbon atoms (C 3-10 cycloalkyl), further preferably 3-7 carbon atoms (C 3-7 cycloalkyl), 4-6 carbon atoms (C 4-6 cycloalkyl), 5-6 carbon atoms (C5-6 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, methylcyclopropyl, 2-ethyl-cyclopentyl, dimethylcyclobutyl, norbornyl (bicyclo[2.2.1]heptyl), bicyclo[2.2.2]octyl, adamantyl, and the like.

[0077] The term "alkyl" refers to a monovalent saturated aliphatic hydrocarbon group including straight-chain or branched-chain groups, preferably containing 1-10 carbon atoms (i.e., C 1-10 alkyl), further preferably containing 1-8 carbon atoms (i.e., C 1-8 alkyl), more preferably containing 1-6 carbon atoms (i.e., C 1-6 alkyl). For example, "C 1-6 alkyl" means that the group is an alkyl group and the number of carbon atoms in the carbon chain is between 1-6 (specifically 1, 2, 3, 4, 5, or 6). Examples include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, sec-butyl, n-pentyl, neopentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, and the like.

[0078] The term "-alkyl-" or "alkylene" refers to a saturated straight-chain or branched-chain divalent hydrocarbon radical. For example, C1-C8 alkylene refers to a straight-chain or branched-chain alkylene group having 1-8 number of carbon atoms.

[0079] The term "alkynyl," unless otherwise specified, refers to a straight-chain or branched-chain unsaturated aliphatic hydrocarbon group having at least one triple bond, consisting of carbon and hydrogen atoms. The alkynyl group can contain 2-20 carbon atoms, preferably 2-10 carbon atoms (i.e., C 2-10 alkynyl), further preferably containing 2-8 carbon atoms (i.e., C 2-8 alkynyl), more preferably containing 2-6 carbon atoms (i.e., C 2-6 alkynyl), 2-5 carbon atoms (i.e., C 2-5 alkynyl), 2-4 carbon atoms (i.e., C 2-4 alkynyl), 2-3 carbon atoms (i.e., C 2-3 alkynyl), 2 carbon atoms (i.e., C2 alkynyl), for example, "C 2-6 alkynyl" means that the group is an alkynyl group and the number of carbon atoms in the carbon chain is between 2-6 (specifically 2, 3, 4, 5, or 6). Non-limiting examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, and 1-butynyl, and the like.

[0080] The term "heteroalkyl" by itself or in combination with another term means a stable straight-chain, branched-chain alkyl radical or combinations thereof, consisting of a number of carbon atoms and at least one heteroatom. The number of carbon atoms can range from 1 to 50 (preferably 1 to 20, more preferably 1 to 12, most preferably 1 to 8), such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50. The heteroalkyl group can optionally contain one, two, or more (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12) heteroatoms selected from N, O, S (or interpreted as optional insertion of heteroatoms into optionally C-C and C-H bonds of the alkyl group). The heteroatoms O, N, and S can be located at any interior position of the heteroalkyl group or at the position where the alkyl group is attached to the rest of the molecule. Examples include, but are not limited to, alkoxy, -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH2-S(O)-CH3, -CH2-CH2-S(O)2-CH3.

[0081] The term "amino acid" refers to a naturally occurring amino acid or a non-naturally occurring amino acid, represented by NH2-C(R'R")-C(=0)OH, where each of R' and R" is independently hydrogen, optionally substituted straight, branched, or cyclic alkyl, alkenyl, or alkynyl having from 1 to 10 carbon atoms, aryl, heteroaryl, or heterocyclyl, or R" and the N-terminal nitrogen atom can combine to form a heterocyclic ring, such as, for example, phenylalanine (F), glycine (G), valine (V), citrulline (C), glutamic acid (E), alanine (A), lysine (K), C1-C6 alkyl substituted lysine (e.g., C1-C3 alkyl substituted lysine, such as methyl substituted lysine, ethyl substituted lysine, n-propyl substituted lysine, isopropyl substituted lysine), and the like.

[0082] The term "amino acid residue" refers to the corresponding residue when one hydrogen atom is removed from the amino terminus and / or one hydroxyl group is removed from the carboxyl terminus of an amino acid, such as, for example, -NH-C(R'R")-C(O)-.

[0083] In the general context, the term "peptide" has the meaning commonly understood in the art and refers to a short chain of amino acid monomers linked by peptide (amide) bonds.

[0084] Unless otherwise specified, the compounds described in this application (including antibody-drug conjugates) also comprise "solvents" or "solvent compounds," where "solvent" or "solvent compound" refers to the physical association of the compound with one or more solvent molecules (organic or inorganic). This physical association includes hydrogen bonding. In some cases, such as when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid, the solvate can be separated. Solvent molecules in a solvate may be present in a regular and / or disordered arrangement. Solvent compounds may contain stoichiometric or non-stoichiometric solvent molecules. "Solvent compound" encompasses both solution phases and separable solvates. Exemplary solvates include, but are not limited to, hydrates, ethanolates, methanolates, and isopropanolates. Solvation methods are well known in the art.

[0085] Unless otherwise specified, the compounds described in this application (including antibody-drug conjugates) also contain "isotope labels," which refer to compounds that can exist in an isotopically traced or enriched form, containing one or more atoms whose atomic weights or mass numbers differ from the atomic weights or mass numbers of the most abundant atoms found in nature. The isotopes can be radioactive or non-radioactive. Commonly used isotopes for labeling are hydrogen isotopes. 2 H and 3 H; Carbon isotopes: 13 C and 14 C; Chlorine isotopes: 35 Cl and 37 Cl; Fluorine isotopes: 18 F; Iodine isotopes: 123 I and 125 I; Nitrogen isotopes: 13 N and 15 N; oxygen isotopes: 15 O, 17 O and 18 O and sulfur isotopes 35 S. These isotope-labeled compounds can be used to study the distribution of pharmaceutical molecules in tissues. Especially 2 H and 13 C, due to their ease of labeling and detection, are more widely used. Substitution of certain heavy isotopes, such as deuterium (2H), can enhance metabolic stability and prolong half-life, thereby achieving a therapeutic advantage through dose reduction. Isotope-labeled compounds are generally synthesized starting from a labeled starting material, using known synthetic techniques similar to those used to synthesize non-isotope-labeled compounds.

[0086] The compounds described herein (including antibody conjugates) also include "stereoisomers" which refer to compounds which have the same chemical constitution but differ in the arrangement of atoms or groups in space. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotamers), geometric isomers (cis / trans), atropisomers, and the like. Mixtures of any of the stereoisomers can be separated into the pure or substantially pure geometric isomers, enantiomers, diastereomers, e.g., by chromatographic and / or fractional crystallization methods.

[0087] The compounds described herein (including antibody conjugates) also include "tautomers" which refer to isomers of different energy which can interconvert by a low energy barrier. If tautomerism is possible (e.g., in solution), a chemical equilibrium of the tautomers can be achieved. For example, prototropic tautomers (also known as proton-shift tautomers) include tautomers which interconvert by the migration of a proton, such as keto-enol isomerization and imine-enamine isomerization. Bond tautomers include tautomers which interconvert by reorganization of bonding electrons.

[0088] The compounds described herein (including antibody conjugates) also include "pharmaceutically acceptable salts" or "pharmaceutically acceptable salts" which refer to pharmaceutically acceptable organic or inorganic salts of compounds (e.g., drugs, drug-linker, or antibody-linker-drug conjugates). The compounds can contain at least one amino, imino, hydroxy, or carboxyl group and thus are capable of forming pharmaceutically acceptable salts with various acids or bases. Exemplary salts include, but are not limited to, sulfate, trifluoroacetate, citrate, acetate, oxalate, hydrochloride, hydrobromide, hydroiodide, nitrate, bisulfate, phosphate, acid phosphate, phosphite, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, potassium, sodium, ammonium, calcium, and the like. Additionally, pharmaceutically acceptable salts have more than one charged atom in the structure. Examples where multiple charged atoms are part of a pharmaceutically acceptable salt can have multiple counterions. For example, a pharmaceutically acceptable salt has one or more charged atoms and / or one or more counterions.

[0089] In a first aspect, the present application provides an anti-TF antibody or antigen binding fragment thereof, the antibody or antigen binding fragment comprising a heavy chain variable region comprising three complementarity determining regions HCDR1, HCDR2, and HCDR3, and a light chain variable region comprising three complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein:

[0090] (1) the amino acid sequence of HCDR1 is set forth in SEQ ID NO. 5 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 5, the amino acid sequence of HCDR2 is set forth in SEQ ID NO. 6 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 6, the amino acid sequence of HCDR3 is set forth in SEQ ID NO. 7 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 7, the amino acid sequence of LCDR1 is set forth in SEQ ID NO. 14 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 14, the amino acid sequence of LCDR2 is set forth in SEQ ID NO. 15 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 15, and the amino acid sequence of LCDR3 is set forth in SEQ ID NO. 16 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 16; or

[0091] (2) the amino acid sequence of HCDR1 is set forth in SEQ ID NO. 20 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 20, the amino acid sequence of HCDR2 is set forth in SEQ ID NO. 21 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 21, and the amino acid sequence of HCDR3 is set forth in SEQ ID NO. 22 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 22, the amino acid sequence of LCDR1 is set forth in SEQ ID NO. 27 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 27, the amino acid sequence of LCDR2 is set forth in SEQ ID NO. 28 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 28, and the amino acid sequence of LCDR3 is set forth in SEQ ID NO. 29 or a sequence with at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 29;

[0092] wherein the HCDR and LCDR sequences are defined according to Kabat.

[0093] In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region is the sequence set forth in any one of SEQ ID NO. 1-4, 18 and 19 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to the sequence set forth in any one of SEQ ID NO. 1-4, 18 and 19.

[0094] In some embodiments of the first aspect, the amino acid sequence of the light chain variable region is the sequence set forth in any one of SEQ ID NO. 11-13 and 24-26 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to the sequence set forth in any one of SEQ ID NO. 11-13 and 24-26.

[0095] In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region is the sequence set forth in any one of SEQ ID NO. 1-4 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to the sequence set forth in any one of SEQ ID NO. 1-4, and the amino acid sequence of the light chain variable region is the sequence set forth in any one of SEQ ID NO. 11-13 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to the sequence set forth in any one of SEQ ID NO. 11-13.

[0096] In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region is the sequence set forth in SEQ ID NO. 18 or 19 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to the sequence set forth in SEQ ID NO. 18 or 19, and the amino acid sequence of the light chain variable region is the sequence set forth in any one of SEQ ID NO. 24-26 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to the sequence set forth in any one of SEQ ID NO. 24-26.

[0097] In some embodiments of the first aspect:

[0098] (1) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 1, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 11; or

[0099] (2) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 1, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 12; or

[0100] (3) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 1, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 13; or

[0101] (4) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 2, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 11; or

[0102] (5) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 2, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 12; or

[0103] (6) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 2, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 13; or

[0104] (7) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 3 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 3, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 11; or

[0105] (8) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 3 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 3, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 12; or

[0106] (9) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 3 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 3, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 13; or

[0107] (10) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 4 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 4, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 11; or

[0108] (11) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 4 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 4, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 12; or

[0109] (12) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 4 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 4, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 13; or

[0110] (13) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 18, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 24 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 24; or

[0111] (14) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 18, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 25 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 25; or

[0112] (15) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 18, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 26 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 26; or

[0113] (16) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 19 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 19, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 24 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 24; or

[0114] (17) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 19 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 19, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 25 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 25; or

[0115] (18) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 19 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 19, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 26 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 26;

[0116] (19) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 36 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 36, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 37 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 37; or

[0117] (20) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 38 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 38, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 39 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 39.

[0118] In some embodiments of the first aspect, the antibody or antigen-binding fragment thereof further comprises a heavy chain constant region having an amino acid sequence of any one of SEQ ID NOs. 8-10 and 23 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to any one of SEQ ID NOs. 8-10 and 23.

[0119] In some embodiments of the first aspect, the antibody or antigen-binding fragment thereof further comprises a light chain constant region having an amino acid sequence of any one of SEQ ID NOs. 17, 30-31 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to any one of SEQ ID NOs. 17, 30-31.

[0120] In some embodiments of the first aspect, the antibody or antigen-binding fragment thereof further comprises a heavy chain constant region and a light chain constant region, wherein:

[0121] the heavy chain constant region has an amino acid sequence of any one of SEQ ID NOs. 8, 9, and 23 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to any one of SEQ ID NOs. 8, 9, and 23, and the light chain constant region has an amino acid sequence of SEQ ID NO. 17 or 30 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17 or 30; or

[0122] the heavy chain constant region has an amino acid sequence of SEQ ID NO. 10 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 10, and the light chain constant region has an amino acid sequence of SEQ ID NO. 31 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 31; or

[0123] the heavy chain constant region has an amino acid sequence of SEQ ID NO. 9 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 9, and the light chain constant region has an amino acid sequence of SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17.

[0124] In some embodiments of the first aspect, the antibody or antigen-binding fragment thereof further comprises a heavy chain constant region and a light chain constant region, wherein:

[0125] (1) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 17;

[0126] (2) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 31 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 31;

[0127] (3) the amino acid sequence of the heavy chain variable region is SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is SEQ ID NO. 9 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 9, and the amino acid sequence of the light chain constant region is SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17;

[0128] (4) the amino acid sequence of the heavy chain variable region is SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is SEQ ID NO. 9 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 9, and the amino acid sequence of the light chain constant region is SEQ ID NO. 31 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 31;

[0129] (5) the amino acid sequence of the heavy chain variable region is SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is SEQ ID NO. 10 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO. 10, and the amino acid sequence of the light chain constant region is SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO. 17;

[0130] (6) the amino acid sequence of the heavy chain variable region is SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is SEQ ID NO. 10 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO. 10, and the amino acid sequence of the light chain constant region is SEQ ID NO. 31 or a sequence having at least 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO. 31;

[0131] (7) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 18, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 24 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 24, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 30;

[0132] (8) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 18, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 25 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 25, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 30;

[0133] (9) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 18, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 26 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 26, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 30;

[0134] (10) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 19 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 19, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 24 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 24, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 30;

[0135] (11) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 19 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 19, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 25 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 25, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 30; or

[0136] (12) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 19 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 19, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 26 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 26, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 30; or

[0137] (13) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0138] (14) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 13, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0139] (15) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 2, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0140] (16) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 2, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0141] (17) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 2, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 13, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0142] (18) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 3 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 3, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0143] (19) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 3 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 3, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0144] (20) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 3 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 3, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 13, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0145] (21) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 4 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 4, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0146] (22) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 4 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 4, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0147] (23) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 4 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 4, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 13, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0148] (24) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 10 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 10, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 31 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 31.

[0149] In some embodiments, the anti-TF antibody comprises two identical heavy chains and light chains.

[0150] In the present application, the CDR sequences are determined according to the Kabat rule, unless otherwise specified.

[0151] In some embodiments of the first aspect, the anti-TF antibody or antigen binding fragment thereof is a murine, chimeric, humanized antibody or antigen binding fragment. In some specific embodiments, the anti-TF antibody or antigen binding fragment thereof is a humanized antibody or antigen binding fragment.

[0152] In some embodiments of the first aspect, the anti-TF antibody or antigen-binding fragment thereof is capable of binding human TF. In some particular embodiments, the antibody or antigen-binding fragment thereof is capable of binding human and cynomolgus monkey TF, but not rat or mouse TF.

[0153] In some embodiments of the first aspect, the antigen-binding fragment is an Fv, scFv, Fab, Fab', or (Fab')2.

[0154] In some embodiments of the first aspect, the anti-TF antibody or antigen-binding fragment thereof has ADCC (antibody-dependent cell-mediated cytotoxicity) activity.

[0155] In some embodiments of the first aspect, the anti-TF antibody or antigen-binding fragment thereof has CDC (complement-dependent cytotoxicity) activity.

[0156] In some embodiments of the first aspect, the anti-TF antibody or antigen-binding fragment thereof has ADCP (antibody-dependent cell-mediated phagocytosis) activity.

[0157] In some embodiments of the first aspect, the anti-TF antibody is a full-length antibody.

[0158] In some embodiments of the first aspect, the anti-TF antibody is a monoclonal antibody.

[0159] In some embodiments of the first aspect, the anti-TF antibody is an IgM, IgD, IgG, IgA, or IgE type antibody. In some particular embodiments, the anti-TF antibody is an IgG type antibody. In some more particular embodiments, the anti-TF antibody is an IgGl type antibody.

[0160] In some embodiments of the first aspect, the anti-TF antibody comprises a light chain constant region of the kappa subtype or the lambda subtype.

[0161] In some embodiments of the first aspect, the anti-TF antibody comprises a human IgGl type heavy chain constant region and a human kappa subtype light chain constant region.

[0162] In some embodiments of the first aspect, the anti-TF antibody comprises a wild-type Fc region.

[0163] In some embodiments of the first aspect, the anti-TF antibody is an antibody of the structure shown in the table below:

[0164] In some embodiments of the first aspect, the anti-TF antibody or antigen-binding fragment thereof is engineered to have enhanced ADCC, CDC, and / or ADCP activity. In some embodiments, the anti-TF antibody comprises an engineered Fc region that results in the antibody having enhanced ADCC, CDC, and / or ADCP effect.

[0165] In some embodiments of the first aspect, the anti-TF antibody or antigen-binding fragment thereof is engineered to have reduced, or even eliminated, ADCC, CDC, and / or ADCP activity. In some embodiments, the anti-TF antibody comprises an engineered Fc region that results in the antibody having reduced, or even eliminated, ADCC, CDC, and / or ADCP effect. In some embodiments, an antibody having reduced, or even eliminated, ADCC, CDC, and / or ADCP activity can be advantageous in view of the target TF and the in vivo application environment of the antibody, as too strong Fc effect can affect or even diminish the therapeutic efficacy of the antibody. Means to reduce or eliminate Fc effect are known in the art, for example, 1) amino acid residue mutation engineering (mainly to reduce binding to relevant receptors), 2) glycosylation engineering, 3) IgG4 type antibody engineering, etc.

[0166] In the present application, as non-limiting examples, the Fc region of the anti-TF antibody can comprise amino acid mutations that reduce ADCC, CDC, and / or ADCP effect.

[0167] In a second aspect, the present application provides an isolated nucleic acid molecule encoding the anti-TF antibody or antigen-binding fragment thereof of the first aspect.

[0168] In a second aspect, the present application also provides a combination of isolated nucleic acid molecules, comprising a nucleic acid molecule encoding the light chain of the antibody or antigen-binding fragment thereof of the present application and a nucleic acid molecule encoding the heavy chain of the antibody or antigen-binding fragment thereof of the present application. In some embodiments, the nucleic acid molecules are operably linked to regulatory sequences, which can be recognized by a host cell transformed with the vector.

[0169] In a third aspect, the present application provides a vector (e.g., an expression vector) comprising the nucleic acid molecule or the combination of nucleic acid molecules of the second aspect.

[0170] In some embodiments, the expression vector of the present application comprises the nucleic acid molecule or the combination of nucleic acid molecules of the present application, operably linked to regulatory sequences that allow the expression of the encoded polypeptide in a host cell or a cell-free expression system. The choice of expression vector depends on the choice of host cell, and can be selected so as to have the desired expression and regulatory characteristics in the chosen host cell.

[0171] The nucleic acid in the vector can be operably linked to one or more expression control sequences. As used herein, "operably linked" means incorporated into a genetic construct in a manner which enables the expression control sequence to control expression of the coding sequence of interest. Examples of expression control sequences include promoters, enhancers, and transcription termination regions. A promoter is an expression control sequence that consists of a DNA molecule from a region that is usually upstream (generally near the start site for RNA polymerase II) of the transcription initiation point. To place a coding sequence under the control of a promoter, the translational start site of the polypeptide-translation reading frame must be positioned between 1 and about 50 nucleotides downstream of the promoter. Enhancers provide expression specificity in terms of time, location, and level. Unlike promoters, enhancers can function when located at various distances from the transcriptional start site. Enhancers can also be located downstream of the transcriptional start site. A coding sequence is "operably linked" to and "under the control" of an expression control sequence in a cell when the RNA polymerase is able to transcribe the coding sequence into mRNA, which can then be translated into a protein encoded by the coding sequence.

[0172] Suitable expression vectors include, but are not limited to, plasmids and viral vectors derived from, for example, bacteriophage, baculovirus, tobacco mosaic virus, herpes virus, cytomegalovirus, retrovirus, vaccinia virus, adenovirus, and adeno-associated virus. Many vectors and expression systems are commercially available from companies such as Novagen (Madison, WI), Clontech (Palo Alto, CA), Stratagene (La Jolla, CA), and Invitrogen Life Technologies (Carlsbad, CA).

[0173] Expression vectors can include a tag sequence. The tag sequence is typically expressed as a fusion to the encoded polypeptide. Such tags can be inserted at any location within the polypeptide, including the carboxyl or amino terminus. Examples of useful tags include, but are not limited to, Fc fragments, polyhistidine, green fluorescent protein (GFP), glutathione S-transferase (GST), c-myc, hemagglutinin, Flag™ tag (Kodak, New Haven, CT), maltose E binding protein, and protein A.

[0174] In a fourth aspect, the present application provides a cell containing the nucleic acid molecule or combination of nucleic acid molecules of the second aspect or the vector of the third aspect.

[0175] In some embodiments, the host cell can be a prokaryotic host cell, a eukaryotic host cell or a bacteriophage. The prokaryotic host cell can be E. coli, B. subtilis, Streptomyces or P. irridescens, etc. The eukaryotic host cell can be a fungus such as P. pastoris, S. cerevisiae, S. pombe, Trichoderma, etc., an insect cell such as Spodoptera frugiperda, etc., a plant cell such as tobacco, etc., a mammalian cell such as BHK cell, CHO cell, COS cell, myeloma cell, etc. In some embodiments, the host cell is preferably a mammalian cell, more preferably a BHK cell, a CHO cell, an NSO cell or a COS cell.

[0176] In a fifth aspect, the present application provides an antibody-drug conjugate comprising the anti-TF antibody or antigen-binding fragment thereof of the first aspect, or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer or isotopically-labeled form thereof.

[0177] In some embodiments, the antibody-drug conjugate has the structure shown in Formula (I):

[0178] A-(L-D) d (I)

[0179] wherein:

[0180] A is the anti-TF antibody or antigen-binding fragment thereof of the first aspect of the present application;

[0181] D is a pharmaceutically active molecule, preferably a molecule having a disease-treating activity, more preferably a molecule having a cancer-inhibiting activity, most preferably a small molecule having a cancer-inhibiting activity;

[0182] L is a linker moiety, one end of which is connected to A and the other end of which is connected to the pharmaceutically active molecule D;

[0183] d is an integer or a decimal number in the range of 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12).

[0184] In some embodiments, the structure of -L- is -L1-L2-L3-L4-, wherein L1 is a covalent linking unit which is covalently linked to A, L2 is an extension unit, L3 is a peptide residue consisting of 2-8 amino acids, and L4 is a bond or a self-cleaving fragment.

[0185] In some embodiments, L1 is selected from wherein * indicates the connection to A.

[0186] In some embodiments, L2 is selected from -L 2a -, 2b -, - C(O)-, -NH-, -SO2-, -CH2-; preferably, L2 is selected from -L2a -, -L 2a -C(O)-, -L 2a -L 2b -C(O)-, -L 2a -NH-C(O)-, -L 2a -L 2b -NH-C(O)-, -L 2a -L 2b -C(O)-NH-, -L 2a -C(O)-NH-L 2b -C(O)-, -L 2a -NH-C(O)-L 2b -C(O)-, -L 2a -C(O)-NH-L 2b -C(O)-NH- or -L 2a -NH-C(O)-L 2b -NH-C(O)-, wherein L 2a , L 2b each independently is selected from -Ci-C8alkylene-, -Ci-C8alkylene-C3-C8cycloalkylene-, -C2-C8alkynylene-, -C6-C 14 arylene-, -C6-C 14 arylene-Ci-C8alkylene-, -5-10 membered heteroarylene-, -5-10 membered heteroarylene-Ci-C8alkylene-, straight chain or branched heteroalkylene of 1-50 carbon atoms, straight chain or branched heteroalkylene-3-8 membered heterocyclyl of 1-50 carbon atoms, each of said alkylene, cycloalkylene, arylene, heteroalkylene, heteroarylene, heterocyclyl being optionally substituted with one or more substituents independently selected from halogen, Ci-C6alkyl, heteroalkyl of 1-6 carbon atoms (such as Ci-C6alkoxy), hydroxyl, amino, carboxyl, or C3-C8cycloalkyl, said heteroalkylene, heterocyclyl, heteroalkyl containing 1-12 heteroatoms, said heteroatoms of said heteroalkylene, heterocyclyl, heteroarylene, heteroalkyl being selected from one or more of N, O, or S.

[0187] In some embodiments, L2is selected from -L 2a -, -L 2a -C(O)-, -L 2a -NH-C(O)-L 2b -C(O)-, wherein L 2a and L 2b each independently is selected from -Ci-C6alkylene-, -Ci-C3alkylene-C3-C6cycloalkylene-, -ethynylene-Ci-C6alkylene-, -(CH2CH2O) n -, -(CH2CH2O)n C 1-3 alkylene-, -C 1-3 alkylene (CH2CH20) n C 1-3 alkylene-, -C 1-3 alkylene-O-C 1-3 alkylene, phenyl, -phenyl-C 1-3 alkylene-, wherein n is an integer selected from 1 to 12 (such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, preferably an integer from 3 to 8).

[0188] In some embodiments, L2is selected from -L2a-, -L2a-C(O)-, -L2a-NH-C(O)-L2b-C(O)-, wherein L2aand L2bare each independently selected from -(CH2)5-, phenylene, -phenyl-CH2-, -CH2-O-CH2-. 2a -C(O)-, -L 2a -NH-C(O)-L 2b -C(O)-, wherein L 2a selected from methylene, ethylene, n-propylene, n-butylene, n-pentylene, n-hexylene, -C1-C3alkylene- cyclohexanediyl-, -phenylene-, -phenylene-C1-C3alkylene-, straight chain or branched heteroalkylene of 1 to 12 carbon atoms; L 2b selected from methylene, ethylene, n-propylene, n-butylene, n-pentylene, n-hexylene, -CH2OCH2-, -CH2CH2OCH2-, -CH2CH2OCH2CH2-, -CH2O-, -CH2CH2O-, -(OCH2CH2) q -, -CH2CH2(OCH2CH2) q -, -CH2(OCH2CH2) q -, -CH2CH2(OCH2CH2) q CH2-, -CH2CH2(OCH2CH2) q CH2CH2-, -CH2(OCH2CH2) q CH2CH2-, wherein q is an integer from 1 to 12.

[0189] In some embodiments, L2is selected from -L2a-, -L2a-C(O)-, -L2a-NH-C(O)-L2b-C(O)-, wherein L2aand L2bare each independently selected from -(CH2)5-, phenylene, -phenyl-CH2-, -CH2-O-CH2-.

[0190] In some embodiments, L2is selected from -L2a-, -L2a-C(O)-, -L2a-NH-C(O)-L2b-C(O)-, wherein L2ais selected from -(CH2)5-, phenylene, and L2bis selected from: -phenyl-CH2-, -CH2-O-CH2-.

[0191] In some embodiments, L2 is selected from: -(CH2CH2O) n CH2C(O)-, -(CH2CH2O) n CH2CH2-, wherein the * position is connected to L1, and n is as defined above.

[0192] In some embodiments, L3 is a peptide residue consisting of 2-8 (such as 2, 3, 4, 5, 6, 7, 8) natural or non-natural amino acids, wherein the amino acids are optionally further substituted with one or more substituents selected from C1-C6 alkyl, heteroalkyl of 2-6 atoms, C1-C6 alkoxy, hydroxyl, amino, carboxyl, or C3-C8 cycloalkyl.

[0193] In some embodiments, L3 is a peptide residue consisting of 2-8 natural or non-natural amino acids selected from the group consisting of: phenylalanine, isoleucine, leucine, tryptophan, valine, methionine, tyrosine, alanine, threonine, histidine, serine, glutamine, arginine, lysine, asparagine, glutamic acid, proline, citrulline, aspartic acid, and glycine (preferably, phenylalanine, valine, alanine, serine, glutamine, arginine, lysine, asparagine, glutamic acid, proline, citrulline, aspartic acid, and glycine, C1-C6 alkyl substituted lysine (such as C1-C3 alkyl substituted lysine, such as methyl substituted lysine, ethyl substituted lysine, n-propyl substituted lysine, isopropyl substituted lysine), and the like.

[0194] In some embodiments, L3 is a peptide residue consisting of 2-8 amino acids selected from the group consisting of phenylalanine, valine, alanine, serine, glutamine, arginine, lysine, asparagine, glutamic acid, proline, citrulline, aspartic acid, and glycine.

[0195] In some embodiments, L3 is selected from -ValCit-; -AlaAla-; -ValAla-; -PheLys-; -AlaAlaAla-; -GluValCit-; -GlyGlyPheGly-.

[0196] In some embodiments, L3 has a structure selected from: wherein the * position is connected to L2.

[0197] In some embodiments, L4 is selected from: -NCH2-, wherein the * position is connected to L3, wherein R1 is selected from hydrogen and wherein R2 is a hydrophilic segment, preferably the hydrophilic segment comprises a polyethylene glycol, a poly natural or non-natural amino acid group, a monosaccharide, an oligosaccharide or a polysaccharide or a combination thereof, more preferably the hydrophilic segment comprises a linear or branched heteroalkyl of 4-50 (such as 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, preferably 4-24, more preferably 6-24, most preferably 8-24) -OCH2CH2- structural units, a peptide chain comprising 4-50 (such as 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, preferably 4-24, more preferably 6-24, most preferably 8-24) proteinogenic amino acids (such as glycine) or non-proteinogenic amino acids (such as sarcosine), or a linear or branched heteroalkyl comprising a monosaccharide, an oligosaccharide or a polysaccharide.

[0198] In some embodiments, L4 is selected from: -NCH2-, wherein the * position is connected to L3, wherein R1 is selected from hydrogen and wherein R2 is selected from:

[0199] wherein r, s, t, u are each independently selected from an integer from 1-50 (such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50). In some embodiments, r is an integer selected from 8-24.

[0200] In some embodiments, R2 has a structure selected from:

[0201] In some embodiments, L4 is selected from -NCH2-,

[0202] In some embodiments, L4is selected from:

[0203] - NCH2-,

[0204] In some embodiments, L has a structure selected from:

[0205] wherein the *position is attached to A. In some specific embodiments, R2is r is an integer from 8 to 24, for example 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24.

[0206] In some embodiments, L has a structure selected from:

[0207] wherein the *position is attached to A. In some specific embodiments, r is an integer from 8 to 24, for example 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24.

[0208] In some embodiments, L is selected from mc-Val-Cit-pABC, mc-Val-Cit, NH2-(PEG)m-Val-Cit, NH2-(PEG)m-Val-Cit-pAB, wherein m is an integer from 1 to 8.

[0209] In some embodiments, D is a molecule having anticancer activity selected from a cytotoxic drug, an immunopotentiator, and a radioisotope.

[0210] In some embodiments, the cytotoxic drug is selected from a microtubulin inhibitor, a DNA topoisomerase inhibitor, a DNA damaging agent, an antimetabolite, or an antitumor antibiotic.

[0211] In some embodiments, the microtubulin inhibitor is selected from an auristatin derivative (e.g., MMAE (Monomethyl auristatin E), MMAF (Monomethyl auristatin F)) or a maytansinoid derivative (e.g., DM1, DM4, Ansamitocin, Mertansine, or dolastatin and derivatives thereof).

[0212] In some embodiments, the DNA topoisomerase inhibitor is selected from camptothecin analogs or DNA topoisomerase I inhibitors and derivatives thereof, for example, DXD, SN38, irinotecan, irinotecan hydrochloride, camptothecin, 9-aminocamptothecin, 9-nitrocamptothecin, 10-hydroxy camptothecin, 9-chloro-10-hydroxy camptothecin, 22-hydroxy camptothecin, topotecan, lurtotecan, belotecan, exatecan, homosilatecan, 6,8-dibromo-2-methyl-3-[2-(D-xylopyranosylamino)phenyl]-4(3H)-quinazolinone, 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-(2E)-2-propenamide, 2-cyano-3-(3,4-dihydroxyphenyl)-N-(3-hydroxyphenylpropyl)-(E)-2-propenamide, 12- -D-xylopyranosyl-12,13-dihydro-2,10-dihydroxy-6-[[2-hydroxy-1-(hydroxymethyl)ethyl]amino]-5H-indolo[2,3-a]pyrrolo[3,4-c]carbazol-5,7(6H)-dione, N-[2-(dimethylamino)ethyl]-4-acridinecarboxamide dihydrochloride, N-[2-(dimethylamino)ethyl]-4-acridinecarboxamide, and derivatives thereof.

[0213] In some embodiments, the DNA damaging agent is selected from calicheamicin class, duocarmycin class, antracycline derivatives PBD (pyrrolobenzodiazepine).

[0214] In some embodiments, the immune enhancer is selected from levamisole, pidotimod, imiquimod, isoprinosine, poliinosinic acid or poliinosinic acid; and / or the antimetabolite is selected from methotrexate, 6-mercaptopurine or 5-fluorouracil; and / or the antitumor antibiotic is selected from polypeptide class antibiotics (such as actinomycin D or bleomycin) or anthraquinones (such as doxorubicin or mitoxantrone).

[0215] In some embodiments, the radioisotope is selected from 211 At、 131 I、 125 I、 90 Y、 186 Re、 188 Re、 153 Sm、 212 Bi、 32 P、 60 Co or 177 Lu.

[0216] In some embodiments, D has the following structure:

[0217] wherein X is selected from a bond or wherein R3, R4are each independently selected from H, C 1-3 alkyl (preferably methyl, ethyl, n-propyl, i-propyl), 3-6 membered cycloalkyl (preferably cyclopropanyl, cyclobutanyl, cyclopentanoyl, cyclohexanoyl), or R3and R4together with the carbon atom to which they are attached form a 3-6 membered cycloalkyl (preferably cyclopropanyl, cyclobutanoyl, cyclopentanoyl, cyclohexanoyl), wherein the * position is attached to L (when the structure of -L- is -L1-L2-L3-L4-, to L4).

[0218] In some embodiments, X is selected from a bond,

[0219] In some embodiments, D has a structure selected from:

[0220] In some embodiments, the antibody-drug conjugate of Formula I has the structure shown below:

[0221] In some embodiments, the antibody-drug conjugate of Formula I has the structure shown below:

[0222] wherein r is an integer selected from 1-50, preferably 8-24, preferably 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; d is an integer or decimal number selected from 1 to 12, preferably 4, 6, or 8.

[0223] In some embodiments, the antibody-drug conjugate of Formula I is selected from the group consisting of hu51-H1L1, hu51-H1L2, hu51-H1L3, hu51-H2L1, hu51-H2L2, hu51-H2L3, hu51-H3L1, hu51-H3L2, hu51-H3L3, hu51-H4L1, hu51-H4L2, hu51-H4L3, hu51-H1L2-6, hu51-H1L2-4, hu52-H1L1, hu52-H1L2, hu52-H1L3, hu52-H2L1, hu52-H2L2, and hu52-H2L3, as numbered in the present application. In some embodiments, the antibody-drug conjugate of Formula I is selected from the group consisting of hu51-H1L2, hu51-H1L3, hu52-H2L1, hu52-H2L3, hu51-H1L2-6, and hu51-H1L2-4, as numbered in the present application.

[0224] In some embodiments, the antibody-drug conjugate of Formula I has a structure selected from the group consisting of:

[0225] In some embodiments, the antibody-drug conjugate of Formula I has a structure selected from the group consisting of:

[0226] wherein A is an anti-TF antibody comprising a heavy chain variable region of amino acid sequence set forth in SEQ ID NO. 1, a light chain variable region of amino acid sequence set forth in SEQ ID NO. 12, a heavy chain constant region of amino acid sequence set forth in SEQ ID NO. 8, and a light chain constant region of amino acid sequence set forth in SEQ ID NO. 17, or an anti-TF antibody comprising a heavy chain variable region of amino acid sequence set forth in SEQ ID NO. 1, a light chain variable region of amino acid sequence set forth in SEQ ID NO. 13, a heavy chain constant region of amino acid sequence set forth in SEQ ID NO. 8, and a light chain constant region of amino acid sequence set forth in SEQ ID NO. 17, or an anti-TF antibody comprising a heavy chain variable region of amino acid sequence set forth in SEQ ID NO. 19, a light chain variable region of amino acid sequence set forth in SEQ ID NO. 24, a heavy chain constant region of amino acid sequence set forth in SEQ ID NO. 23, and a light chain constant region of amino acid sequence set forth in SEQ ID NO. 30, or an anti-TF antibody comprising a heavy chain variable region of amino acid sequence set forth in SEQ ID NO. 19, a light chain variable region of amino acid sequence set forth in SEQ ID NO. 26, a heavy chain constant region of amino acid sequence set forth in SEQ ID NO. 23, and a light chain constant region of amino acid sequence set forth in SEQ ID NO. 30, and d is an integer or decimal number selected from 1 to 12, preferably 4, 6, or 8.

[0227] In some embodiments, the antibody-drug conjugate of Formula I has a structure selected from the group consisting of:

[0228] wherein A is an anti-TF antibody comprising a heavy chain variable region of amino acid sequence set forth in SEQ ID NO. 1, a light chain variable region of amino acid sequence set forth in SEQ ID NO. 12, a heavy chain constant region of amino acid sequence set forth in SEQ ID NO. 8, and a light chain constant region of amino acid sequence set forth in SEQ ID NO. 17, or an anti-TF antibody comprising a heavy chain variable region of amino acid sequence set forth in SEQ ID NO. 1, a light chain variable region of amino acid sequence set forth in SEQ ID NO. 12, a heavy chain constant region of amino acid sequence set forth in SEQ ID NO. 9, and a light chain constant region of amino acid sequence set forth in SEQ ID NO. 17, and d is an integer or decimal number selected from 1 to 12, preferably 4, 6, or 8.

[0229] In some embodiments, the antibody-drug conjugate of Formula I has a structure selected from the group consisting of:

[0230] wherein A is an anti-TF antibody comprising a heavy chain variable region of an amino acid sequence as set forth in SEQ ID NO. 1, a light chain variable region of an amino acid sequence as set forth in SEQ ID NO. 12, a heavy chain constant region of an amino acid sequence as set forth in SEQ ID NO. 9, and a light chain constant region of an amino acid sequence as set forth in SEQ ID NO. 17, or an anti-TF antibody comprising a heavy chain variable region of an amino acid sequence as set forth in SEQ ID NO. 1, a light chain variable region of an amino acid sequence as set forth in SEQ ID NO. 12, a heavy chain constant region of an amino acid sequence as set forth in SEQ ID NO. 10, and a light chain constant region of an amino acid sequence as set forth in SEQ ID NO. 31, and d is an integer or decimal number selected from 1 to 12, preferably 4, 6 or 8.

[0231] In a sixth aspect, the present application provides a pharmaceutical composition comprising the anti-TF antibody or antigen binding fragment thereof of the first aspect, or the antibody-drug conjugate or the pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer or isotopically-labeled of the antibody-drug conjugate of the fifth aspect, and a pharmaceutically acceptable carrier.

[0232] In a seventh aspect, the present application provides use of the anti-TF antibody or antigen binding fragment thereof of the first aspect, or the antibody-drug conjugate or the pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer or isotopically-labeled of the antibody-drug conjugate of the fifth aspect, in the manufacture of a medicament for treating and / or preventing a TF-mediated disease or disorder.

[0233] In an eighth aspect, the present application provides a method of treating and / or preventing a TF-mediated disease or disorder in an individual, comprising administering to an individual in need thereof an effective amount of the anti-TF antibody or antigen binding fragment thereof of the first aspect, or the antibody-drug conjugate or the pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer or isotopically-labeled of the antibody-drug conjugate of the fifth aspect, or the pharmaceutical composition of the sixth aspect.

[0234] In a ninth aspect, the present application provides a pharmaceutical combination comprising the anti-Tissue Factor (TF) antibody or antigen binding fragment thereof of the first aspect, or the antibody-drug conjugate or the pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer or isotopically-labeled of the antibody-drug conjugate of the fifth aspect, or the pharmaceutical composition of the sixth aspect, and one or more additional therapeutic agents.

[0235] In a tenth aspect, the present application provides a kit comprising the anti-tissue factor (TF) antibody or antigen-binding fragment thereof of the first aspect, or the antibody-drug conjugate or the pharmaceutical salt, hydrate, solvate, tautomer, stereoisomer or isotopically-labeled of the fifth aspect, or the pharmaceutical composition of the sixth aspect.

[0236] In some embodiments, the kit comprises a container holding the anti-tissue factor (TF) antibody or antigen-binding fragment thereof of the first aspect, or the antibody-drug conjugate or the pharmaceutical salt, hydrate, solvate, tautomer, stereoisomer or isotopically-labeled of the fifth aspect, or the pharmaceutical composition of the sixth aspect, and optionally comprises a package insert.

[0237] In a tenth aspect, the present application provides a kit comprising the anti-tissue factor (TF) antibody or antigen-binding fragment thereof of the first aspect, or the antibody-drug conjugate or the pharmaceutical salt, hydrate, solvate, tautomer, stereoisomer or isotopically-labeled of the fifth aspect, or the pharmaceutical composition of the sixth aspect.

[0238] The indications related to the sixth to eleventh aspects are described together below.

[0239] In some embodiments, the TF-mediated disease or disorder is cancer.

[0240] In some embodiments, the cancer is a TF-positive cancer, i.e., a cancer in which cancer cells express TF (TF+) on their surface.

[0241] In some embodiments, the cancer is a cancer in which cancer cells express TF (TF+) on their surface at a high level.

[0242] In some embodiments, the expression level of an antigen in a cancer can be characterized according to the percentage of cancer cells that are positive for the antigen. For example, a cancer that expresses TF (TF+) at a high level can mean that at least 60% of cancer cells in the cancer cell population express TF, or at least 70% of cancer cells express TF, or at least 80% of cancer cells express TF, or at least 90% of cancer cells express TF, or at least 95% of cancer cells express TF, or at least 98% of cancer cells express TF, or at least 99% of cancer cells express TF.

[0243] In other embodiments, the degree of TF positivity of a tumor can be qualitatively, quantitatively or semi-quantitatively analyzed according to various methods in the art.

[0244] In some embodiments, the cancer is a solid tumor.

[0245] In some embodiments, the cancer is a hematological neoplasm.

[0246] In some embodiments, the cancer is cervical cancer, pancreatic cancer, head and neck cancer (e.g., pharyngeal squamous carcinoma), endometrial cancer, esophageal cancer, prostate cancer, non-small cell lung cancer, or ovarian cancer.

[0247] In other aspects, the present application provides an anti-TF antibody, wherein the anti-TF antibody comprises a heavy chain variable region, a heavy chain constant region, a light chain variable region, and a light chain constant region, wherein:

[0248] (1) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 17; or

[0249] (2) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 31 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 31; or

[0250] (3) the amino acid sequence of the heavy chain variable region is SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is SEQ ID NO. 9 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 9, and the amino acid sequence of the light chain constant region is SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0251] (4) the amino acid sequence of the heavy chain variable region is SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is SEQ ID NO. 9 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 9, and the amino acid sequence of the light chain constant region is SEQ ID NO. 31 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 31; or

[0252] (5) the amino acid sequence of the heavy chain variable region is SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is SEQ ID NO. 10 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 10, and the amino acid sequence of the light chain constant region is SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0253] (6) the amino acid sequence of the heavy chain variable region is SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is SEQ ID NO. 10 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 10, and the amino acid sequence of the light chain constant region is SEQ ID NO. 31 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 31; or

[0254] (7) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 18, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 24 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 24, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 30; or

[0255] (8) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 18, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 25 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 25, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 30; or

[0256] (9) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 18, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 26 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 26, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 30; or

[0257] (10) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 19 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 19, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 24 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 24, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 30; or

[0258] (11) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 19 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 19, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 25 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 25, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 30; or

[0259] (12) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 19 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 19, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 26 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 26, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 30; or

[0260] (13) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0261] (14) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 13, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0262] (15) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 2, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0263] (16) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 2, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0264] (17) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 2, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 13, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0265] (18) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 3 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 3, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0266] (19) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 3 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 3, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0267] (20) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 3 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 3, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 13, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0268] (21) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 4 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 4, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0269] (22) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 4 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 4, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0270] (23) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 4 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 4, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 13, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; or

[0271] (24) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 10 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 10, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 31 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 31.

[0272] In some embodiments, the anti-TF antibody comprises two identical heavy chains and light chains.

[0273] In other aspects, the present application provides an anti-TF antibody or antigen binding fragment thereof, comprising a heavy chain variable region comprising three complementarity determining regions HCDR1, HCDR2, and HCDR3, wherein:

[0274] (1) the amino acid sequence of HCDR1 is set forth in SEQ ID NO. 5 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 5, the amino acid sequence of HCDR2 is set forth in SEQ ID NO. 6 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 6, and the amino acid sequence of HCDR3 is set forth in SEQ ID NO. 7 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 7; or

[0275] (2) the amino acid sequence of HCDR1 is set forth in SEQ ID NO. 20 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 20, the amino acid sequence of HCDR2 is set forth in SEQ ID NO. 21 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 21, and the amino acid sequence of HCDR3 is set forth in SEQ ID NO. 22 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 22.

[0276] In other aspects, the present application provides an anti-TF antibody or antigen binding fragment thereof, comprising a heavy chain variable region comprising three complementarity determining regions, HCDR1, HCDR2, and HCDR3, wherein:

[0277] (1) the amino acid sequence of HCDR1 is set forth in SEQ ID NO. 5 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 5, the amino acid sequence of HCDR2 is set forth in SEQ ID NO. 6 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 6, and the amino acid sequence of HCDR3 is set forth in SEQ ID NO. 7 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 7; or

[0278] (2) the amino acid sequence of LCDR1 is set forth in SEQ ID NO. 27 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 27, the amino acid sequence of LCDR2 is set forth in SEQ ID NO. 28 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 28, and the amino acid sequence of LCDR3 is set forth in SEQ ID NO. 29 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 29.

[0279] In other aspects, the present application provides an anti-TF antibody or antigen binding fragment thereof, wherein the anti-TF antibody or antigen binding fragment thereof comprises a heavy chain variable region, wherein:

[0280] (1) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 1;

[0281] (2) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 2;

[0282] (3) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 3 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 3;

[0283] (4) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 4 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 4;

[0284] (5) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 18; or

[0285] (6) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 19 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 19.

[0286] In other aspects, the application provides an anti-TF antibody or antigen-binding fragment thereof, wherein the anti-TF antibody or antigen-binding fragment thereof comprises a heavy chain variable region, wherein:

[0287] (1) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 1;

[0288] (2) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 2;

[0289] (3) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 3 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 3;

[0290] (4) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 24 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 24;

[0291] (5) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 25 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 25; or

[0292] (6) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 26 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 26.

[0293] In other aspects, the application provides an anti-TF antibody or antigen-binding fragment thereof, the anti-TF antibody or antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein:

[0294] (i) the amino acid sequence of the heavy chain variable region is the sequence set forth in any one of SEQ ID NO. 1-4 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in any one of SEQ ID NO. 1-4, and the amino acid sequence of the light chain variable region is the sequence set forth in any one of SEQ ID NO. 11-13 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in any one of SEQ ID NO. 11-13; or

[0295] (ii) the amino acid sequence of the heavy chain variable region is the sequence set forth in SEQ ID NO. 18 or 19 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 18 or 19, and the amino acid sequence of the light chain variable region is the sequence set forth in any one of SEQ ID NO. 24-26 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in any one of SEQ ID NO. 24-26.

[0296] It should be understood that the embodiments and technical features described in relation to the anti-TF antibody or antigen binding fragment of the first aspect apply equally to the anti-TF antibody or antigen binding fragment described in the other aspects above, insofar as there is no contradiction. It should also be understood that the nucleic acid molecules, vectors, cells, antibody-drug conjugates, pharmaceutical compositions, uses, methods, pharmaceutical combinations, kits described above in relation to the anti-TF antibody or antigen binding fragment of the first aspect apply equally to the anti-TF antibody or antigen binding fragment described in the other aspects above, insofar as there is no contradiction.

[0297] It should be understood that the aspects and embodiments of the present application described herein include “comprising”, “consisting of”, and “consisting essentially of” aspects and embodiments. The embodiments of the present application are described in detail above, but the present application is not limited thereto. Various simple modifications can be made to the technical solutions of the present application within the technical concept of the present application, including the combination of various technical features in any other suitable manner, and these simple modifications and combinations should also be considered as disclosed by the present application and fall within the protection scope of the present application.

[0298] It is to be understood that the specific embodiments described above and the examples described below are for purposes of illustration only and are not intended to limit the scope of the present application. The present application includes various aspects, embodiments and combinations of those aspects and / or embodiments. The above description and the following examples are intended to illustrate but not limit the scope of the present application. Other aspects, improvements and modifications that are within the scope of the present application will be apparent to those skilled in the art. Thus, it should be appreciated that the scope of the present application also includes the described improvements and modifications to the described aspects and embodiments. Examples

[0299] Example 1: Preparation of Anti-TF murine monoclonal antibody

[0300] Two strains of BALB / c and SJL mice were selected, each with 5 mice, and immunized with human TF membrane outer region recombinant protein-his tag (TF-his, ACRO item number: TF3-H52H5). The antigen was prepared with complete Freund's adjuvant (Sigma) at a volume ratio of 1:1. According to the immunization process, the immunized animals were bled and serological titer detection was performed. Based on the serological titer detection results, considering the affinity of human and monkey, two SJL mice were selected for hybridoma fusion, and before hybridoma fusion, the last boost was performed.

[0301] Table 1 Mouse immunization scheme (immunization dose based on antigen peptide calculation)

[0302] The mouse spleen cells were mixed with SP2 / 0 mouse myeloma cells at a ratio of 1:20, and cell electrofusion was performed. After cell fusion, the fused cells were mixed with HAT medium to prepare a cell suspension, and then the cell suspension was plated into 96-well cell culture plates using a multi-channel pipette. Each mouse's cell suspension was plated into 30 cell culture plates, and two mice's cell suspension was plated into 60 cell culture plates. The fused cell culture plates were placed in a 5.5% carbon dioxide incubator at 37°C for constant temperature culture.

[0303] After 7 days of culture, positive clones against the target protein were screened by ELISA and FACS methods. According to the ELISA and FACS results, 20 positive clones were selected by limited dilution method to subclone the positive clones to obtain positive, stable and single hybridoma cells. The obtained hybridoma cell strains were cultured in a shake flask to express and purify the antibodies, and the purified samples were detected by ELISA and FACS. The antibodies 51E7D9 and 52G1D7 with high affinity were selected for variable region sequencing and humanization.

[0304] Table 2 Amino acid sequences of murine antibodies 51E7D9 and 52G1D7

[0305] ELISA experimental method:

[0306] ① Sampling: the hybridoma fusion cell supernatant to be detected or the murine antibody expressed and purified after subcloning is sucked into the detection plate for use;

[0307] ② Antigen coating: coat TF protein (ACRO item number: TF3-H52H5), coating concentration is 0.5 μg / ml, 100 μl / well, coating at 37°C for 1 h.

[0308] ③ Add primary antibody: add hybridoma fusion cell supernatant or murine antibody expressed and purified after subcloning;

[0309] ④ Add secondary antibody: select HRP-labeled secondary antibody Perxidase-affiniPure Goat Anti-Mouse IgG, Fcγ Fragment Specific (min X Hu, Bov, Hrs Sr Prot); secondary antibody concentration is 0.25 μg / ml, react at 37°C for 0.5 h;

[0310] ⑤ Color development: add 100 μl color developing solution, react at 25°C for 15 min.

[0311] ⑥ Termination: add 50 μl 1M HCl to terminate the reaction.

[0312] ⑦ Reading: use an enzyme label instrument (Thermo) to read OD450nm reading.

[0313] FACS binding experiment method:

[0314] ① The CHO-K1 cells stably expressing human TF (human TF-CHO-K1 GenScript PROBIO) are treated with a digestion reagent for 1-2 min until the cells fall off, 5 ml of fresh culture medium is added and the culture bottle is blown, then the cells are transferred to a 15 ml centrifuge tube, centrifuged at 300g for 3 min, and the supernatant is discarded. Wash the cells with PBS at 300g for 3 min twice, add 50 μl of cells to each well of a 96-well plate, and the cell content is 1×10 5 cells / well.

[0315] ② Gradient dilute the hybridoma parent clone cell supernatant or the murine antibody expressed and purified after subcloning, then incubate at 4°C for 40 min, wash twice with 150 μl of PBS at 300g for 3 min;

[0316] ③ Add 100 μl of fluorescent secondary antibody Alexa Fluor 647 AffiniPure Goat Anti-Mouse IgG, Fcγ fragment specific (min X Hu, Bov, Hrs Sr Prot) (Jaskson, 115-605-071) to each well, incubate on 4°C constant temperature oscillator for 30 min, remove the 96-well plate, add 150 μl of PBS to each well, centrifuge 1-2 times, and discard the supernatant.

[0317] ④ Use a flow cytometer (FACS CantoII) to read the fluorescence signal value.

[0318] Example 2: Humanization of anti-TF murine monoclonal antibody

[0319] The method of CDR grafting was used to humanize the murine antibody sequence. First, the human germline sequence with the highest homology to the original murine sequence was found as a template by the conventional BLAST method; the CDR of the murine antibody was grafted onto the human template; and the FR amino acids in the antibody that could maintain their original conformation were analyzed according to the structure, and the corresponding amino acids in the antibody were back-mutated to murine amino acids to maintain the original affinity.

[0320] 51E7D9 design obtained four heavy chain variable region sequences H1, H2, H3, H4 and three light chain variable region sequences L1, L2, L3 (see Table 3), which were combined with the above four heavy chain variable region sequences and the above three light chain variable region sequences, respectively, using the hu51-CH1' heavy chain constant region sequence and the hu51-CL1' light chain constant region sequence, to obtain 12 humanized antibody sequences (see Table 5). The hu51-CH1' wild-type heavy chain constant region sequence was mutated to obtain heavy chain constant region sequences hu51-CH2' and hu51-CH3', and hu51-CL1' was a kappa (K) type light chain constant region, and hu51-CL2' was obtained by mutation on the basis of a lambda (L) type light chain constant region. The heavy chain constant region sequence hu51-CH2' obtained by mutation and the heavy chain variable region sequence H1, the light chain variable region sequence L2, and the unmutated light chain constant region sequence hu51-CL1' were combined to obtain one humanized antibody sequence hu51-H1L2-6 (see Table 5). The heavy chain constant region sequence hu51-CH3' obtained by mutation and the light chain constant region sequence hu51-CL2' obtained by mutation were combined with the heavy chain variable region sequence H1 and the light chain variable region sequence L2 to obtain one humanized antibody sequence hu51-H1L2-4 (see Table 5).

[0321] 52G1D7 designed two heavy chain variable region sequences H1, H2 and three light chain variable region sequences L1, L2, L3 (see Table 4), respectively combined with the two heavy chain variable region sequences and the three light chain variable region sequences described above, and respectively linked with hu52-CH' heavy chain constant region sequence and hu52-CL' light chain constant region sequence, to obtain six humanized antibody sequences (see Table 5).

[0322] The following examples of the present application use the foregoing antibodies.

[0323] Table 3 51E7D9 humanized antibody sequence

[0324] Table 4 52G1D7 humanized antibody sequence

[0325] Table 5 Humanized antibody sequence

[0326] Example 3: Expression and purification of humanized monoclonal antibodies

[0327] The heavy chain and light chain sequences of the humanized antibodies described above and the Xiangshen antibodies (TF011 and XB002) were gene synthesized, and after codon optimization, were constructed into PTT5 vectors. After direct synthesis, CHO cells were transfected by electroporation, and after 3-7 days of culture, the supernatant was collected by centrifugation. The MabSelect Sure affinity filler was used for purification, and the purified antibodies were ultrafiltrated and replaced into PBS buffer. The concentration was determined using a NanoDrop instrument, and the antibodies were stored at -20°C.

[0328] Table 6 Xiangshen antibody TF011, XB002 antibody sequence

[0329] Example 4: Affinity evaluation of humanized antibodies

[0330] ELISA:

[0331] Human TF-his protein (ACRO, Cat: TF3-H52H5) was coated on 96-well plates, 2% BSA (bovine serum albumin) was used for blocking at 37°C for 2h, then gradient-diluted antibodies (5μg / ml start, 6-fold dilution, 11 concentration points) were added, after incubation at 37°C for 2h, goat anti-human IgG (H+L) cross-adsorbed secondary antibody (Thermo) was added and incubated at 37°C for 1h, then TMB substrate (3,3',5,5'-tetramethylbenzidine) was added for color development. After stopping the color development reaction, the absorbance value at 450nm was read by a microplate reader and the curve was fitted by GraphPad software.

[0332] FACS:

[0333] Human breast cancer cells MDA-MB-231 positive for TF antigen were collected by trypsin digestion, 1×10 5 cells per well were plated in a 96-well plate, gradient-diluted antibodies (15μg / ml start, 4-fold dilution, 11 concentration points) were added, after incubation at 37°C for 2h, PBS was used for washing, goat anti-human IgG (H+L) cross-adsorbed secondary antibody was added and incubated at 37°C for 1h, then the absorbance value was detected by flow cytometry and the curve was fitted by GraphPad software.

[0334] Table 7 Affinity evaluation of humanized antibodies

[0335] As shown in the affinity data of Table 7, each humanized antibody of the present application can well bind to TF protein at the protein level and cell level, and the affinity of multiple antibodies is comparable to that of positive control antibodies TF011, XB002.

[0336] Example 5: Thermal stability evaluation of humanized monoclonal antibodies

[0337] The differential scanning calorimeter (manufacturer: Malvern, model: PEAQ) was used to detect the stability of the humanized anti-TF antibody. The thermal stability of the molecule was analyzed by differential scanning calorimetry (DSC). Before detection, the sample concentration was determined using Nano Drop, and the sample concentration was diluted to about 1 mg / mL using 1x PBS pH 7.0 buffer. The sample was filtered using a 0.22 μm filter, and 350 μL of the sample was added to the sample well. The parameter settings were as follows: scanning start temperature: 20°C, scanning end temperature: 100°C, scanning rate: 90°C / h. The detection results are shown in the following table. The TOnset (melting point temperature) results of the candidate humanized antibodies of the present application were about 10°C higher than that of TF011, and were comparable to that of XB002, indicating that the candidate antibodies had good thermal stability.

[0338] Table 8 DSC detection results of humanized antibodies

[0339] Example 6: FXa factor generation experiment

[0340] The FXa factor generation experiment of the present example includes the following steps:

[0341] (1) Preparation of antigen-antibody complex sample 1: Mix TF-his protein (ACRO catalog number: TF3-H52H5, 10 μg / ml) with gradient-diluted humanized anti-TF antibodies of the present application (starting at 500 μg / ml, 2-fold gradient dilution) at a ratio of 1:1, 25 μl of TF protein and humanized antibody each, mix well, and then add to a 96-well plate and incubate at 37°C for 60 minutes.

[0342] (2) Preparation of PCPS complex sample 2: Prepare PCPS mixture (EGG PC, phosphatidylcholine, Avanti; DOPS, phosphatidylserine, Avanti) at a ratio of 4:1 by volume, then add 50 μl of PCPS complex sample to the antigen-antibody complex 96-well plate sample well, and incubate at 37°C for 60 minutes.

[0343] (3) Dilute FVII and FX factors (ERL) to 4nM and 400nM respectively, mix at a ratio of 1:1 by volume, mix the FVII and FX factor mixture after mixing with the mixture after step (2) at a ratio of 1:1 (FVII and FX factor mixture: step (2) mixture = 50 μl: 50 μl), then add to the 96-well plate, and incubate at 37°C for 2 hours.

[0344] (4) Mix the sample from step (3) above with stop solution (50 mM HEPES, 100 mM Nacl, 60 mM EDTA, 0.1% PEG8000, 1 mg / ml BSA) 1:1, take 50 μl for each sample. Add 30 μl of 2 mM S2765 (Asnail) to each well, develop color at 37°C, and the developing time is 1 hour.

[0345] (5) Detect the absorbance using a microplate reader (450 nm).

[0346] The experimental results are shown in Figure 1. The antibodies hu51-H1L2 and hu51-H1L3 of the present application do not affect the generation of FXa, indicating that they do not affect the binding of TF and FVII after binding to TF, thereby exerting the coagulation function. The XB002 antibody has a weak effect on the generation of FXa, and the TF011 antibody significantly affects the generation of FXa, indicating that the TF011 antibody has a certain inhibitory effect on the physiological coagulation function of TF.

[0347] Example 7: Species cross study of humanized antibodies

[0348] Human TF-his protein (ACRO item number: TF3-H52H5), cynomolgus monkey TF-his protein (ACRO item number: TF3-C52H3), rat TF-his protein (Sino Biological item number: 80407-R08H), and mouse TF-his protein (ACRO item number: TF3-M52H3) were coated on 96-well plates, respectively, and 2% BSA (bovine serum albumin) was used for blocking at 37°C for 2 hours. Gradient-diluted anti-TF antibodies (5 μg / ml starting, 6-fold dilution, 11 concentration points) were added, and after incubation at 37°C for 2 hours, goat anti-human IgG (H+L) cross-adsorbed secondary antibody (Thermo) was added and incubated at 37°C for 1 hour. TMB substrate (3,3',5,5'-tetramethylbenzidine) was added for color development. After stopping the color development reaction, the absorbance at 450 nm was read using a microplate reader, and the curve was fitted using GraphPad software.

[0349] The experimental results are shown in Table 9. The antibodies hu51-H1L2, hu51-H1L3, hu52-H2L1, and hu52-H2L3 of the present application have comparable affinity for human and cynomolgus monkey TF proteins, and do not bind to rat or mouse TF proteins.

[0350] Table 9: Species cross detection results of humanized antibodies

[0351] Example 8: Effect of antibodies on coagulation activity

[0352] Humanized anti-TF antibodies were diluted with HEPES (hydroxyethylpiperazine ethane sulfonic acid) containing CaCl2, adjusting the final concentration of CaCl2 to 5 mM and the final concentration of antibody to 20 mg / mL, and then incubated with 5 x 10 4 MDA-MB-231 cells positive for TF at 37°C for 30 minutes, and then 50 μL of human plasma was added, mixed quickly, and immediately read continuously at 405 nm (TECAN) to calculate the anticoagulant effect of the anti-TF antibody on the initiation of blood coagulation by the cell surface TF. The results were imported into GraphPad for plotting.

[0353] The experimental results are shown in Figure 2. The hu51-H1L2 did not substantially affect the occurrence of the blood coagulation reaction, and the TF011 antibody prolonged the blood coagulation time, having a certain effect on the physiological blood coagulation effect of TF.

[0354] Example 9: Endocytosis activity study

[0355] The hu51-H1L2, TF011, and XB002 antibodies were respectively incubated with human breast cancer cells MDA-MB-231 in a refrigerator at 4°C for 2 hours, and after the co-incubation, the cells were washed twice with PBS, added with DMEM medium, and then placed in a 37°C, 5% CO2 carbon dioxide incubator for incubation for 6 h, 9 h, 24 h, 30 h, and 48 h, respectively. The cells were harvested at the end of the indicated incubation time. The cells were then stained with Alexa Fluor 488 fluorescent dye-labeled goat anti-human antibody, and incubated at 4°C in the dark for 60 minutes. After the incubation, the cells were washed twice with experimental buffer (PBS buffer containing 2% fetal bovine serum), resuspended in the aforementioned buffer, and then detected by flow cytometry, and the median value of the fluorescent signal in the corresponding wells in the experimental plate was read by a plate reading.

[0356] Endocytosis rate % = (initial average median fluorescence value - average median fluorescence value at different time points) / initial average median fluorescence value x 100%.

[0357] The experimental results are shown in Figure 3 and Table 10. After the antibody hu51-H1L2 of the present application was combined with MDA-MB-231 cells, it was quickly endocytosed into the cells, and the endocytosis rate was as high as 66% at 6 hours, which was significantly higher than that of TF011 and XB002.

[0358] Table 10. Endocytosis rates of hu51-H1L2 and positive control antibodies at different time points

[0359] Example 10: Preparation of linker-drug compounds (LD)

[0360] Compounds LD1 and LD2 (reference WO2023088382A1) were prepared according to known methods or purchased commercially, compound LD3 was prepared according to the following method:

[0361] Explanation of abbreviations

[0362] 1. Preparation of intermediate compound 8

[0363] Compound 1 (47.5 g, 189 mmol) and methanol (250 mL) were added to a reaction flask, 80% hydrazine hydrate (35.4 g, 567 mmol) was slowly added at room temperature, and the reaction was heated to 70 °C and refluxed for 6 hours. White crystals were precipitated after cooling, and the remaining solid was washed with methanol (20 mL x 3) to obtain white solid 2 (47.47 g, yield 100%). MS (ESI): (M+H) + Calculated value 252.1, experimental value 252.2.

[0364] Compound 2 (47.4 g, 189 mmol), potassium hydroxide (12.7 g, 227 mmol) and ethanol (400 mL) were added to a reaction flask and stirred to dissolve at room temperature. After slowly adding carbon disulfide (17 g, 283 mmol), the reaction was heated to 100 °C and refluxed for 5 hours. After removing the solvent under reduced pressure, water (50 mL) was added, the pH was adjusted to 6 with dilute hydrochloric acid, and the organic phase was extracted with dichloromethane (100 mL x 3), combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography to obtain compound 3 (49.9 g, yield 90%). MS (ESI): (M+H) + Calculated value 294.1, experimental value 294.3.

[0365] Compound 3 (5.86 g, 20 mmol), triethylamine (2.42 g, 24 mmol) and tetrahydrofuran (36 mL) were added to a reaction flask, and methyl iodide (3.12 g, 22 mmol) was added to the above reaction solution. The reaction was completed after stirring at 25 °C for 1.5 hours, the solvent was removed under reduced pressure, and the residue was purified by silica gel chromatography to obtain compound 4 (4.05 g, yield 66%). MS (ESI): (M+H) + Calculated value 308.1, experimental value 308.2.

[0366] Compound 4 (3.3 g, 10.6 mmol) and ethyl acetate (20 mL) were added to a reaction flask, and hydrogen chloride ethyl acetate solution (2.7 mL, 4M) was added dropwise at 25 °C, and the reaction was continued for 6 hours. After removing the solvent under reduced pressure, crude product 5 was obtained and directly used in the next step.

[0367] The crude product 5, diethylene glycol anhydride 6 (1.35 g, 11.7 mmol), triethylamine (2.14 g, 21.2 mmol) and tetrahydrofuran (30 mL) were added into a reaction flask, and reacted at 25 °C for 1.5 h. The solvent was removed under reduced pressure, and the residue was added into ether. After filtration, the compound 7 (3.4 g, yield 99%) was obtained after washing with water and ether. MS (ESI): (M+H) + Calculated 324.1, found 324.2.

[0368] The compound 7 (3.4 g, 10.5 mmol) and glacial acetic acid (20 mL) were added into a reaction flask. After dissolution, potassium permanganate (2.48 g, 15.7 mmol) was added at 0 °C. The reaction was resumed at 25 °C for 1 h. Saturated sodium sulfite solution was added until the solution became colorless. The organic phase was extracted with ethyl acetate (100 mL x 3), combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography to obtain the compound 8 (2.4 g, yield 65%). MS (ESI): (M+H) + Calculated 356.0, found 356.0.

[0369] 2. Preparation of compound LD3

[0370] At 0 °C, 1-(4-aminophenyl)-3-butyne-1-ol (4997 mg, 31 mmol), N-fluorenylmethoxycarbonyl-L-alanine (9651 mg, 31 mmol), EEDQ (11498 mg, 46.5 mmol) and super dry DCM (90 mL) were added into a reaction flask. After 3 h of reaction, the solvent was removed under reduced pressure. The residue was slurried with 500 mL MTBE (methyl tert-butyl ether) to obtain the compound 10 (9007 mg, yield 64%, dr = 1:1). MS (ESI): (M+H) + Calculated 455.2, found 455.2.

[0371] Chiral resolution of intermediate 10: Compound 10 (50 g) was resolved by supercritical fluid chromatography (SFC) to obtain compound 11 (19 g, retention time 13.55 min) and compound 12 (20 g, retention time 16.29 min). SFC resolution method: Column type: DAICEL CHIRALCEL OD (250 mm-50 mm, 10 um); Mobile phase: A: CO2, B: CO2-ACN / i-PrOH (0.1% NH3H2O); Isocratic elution: B in A for 50%; Flow rate: 200 mL / min; Detector: PDA; Column temperature: 25 °C; Back pressure: 100 Bar;

[0372] HPLC method: Instrument information: Thermo liquid chromatograph (ADC-U3000-01); column: XBridge BEH C18 (4.6*150mm, 5um); column temperature: 25°C; injection tray temperature: 25°C; mobile phase: A: 10mM NH4FA; B: ACN; flow rate: 0.8mL / min; detection wavelength: 254nm; injection volume: 2ul;

[0373] Gradient condition:

[0374] The structure and configuration of compound 12 were confirmed by X-RAY, and the single crystal diffraction pattern thereof is shown in FIG. 4A and FIG. 4B.

[0375] Compound 12 (3632mg, 8mmol), 90mL THF (commercially available) were added to the reaction bottle at 0°C. Stirring and slowly adding 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) (1215mg, 8mmol), the reaction was removed from the reaction bottle after half an hour. After monitoring the disappearance of the raw material by TLC, the solvent was removed under reduced pressure, and the residue was purified by silica gel chromatography (DCM / MeOH 95:5) to obtain compound 13 (1707mg, yield 92%). MS (ESI): (M+H) + Calculated value 233.1, experimental value 233.2.

[0376] Compound 13 (1624mg, 7mmol), Fmoc-Val-OSu (362mg, 8.4mmol) and N,N-dimethylformamide (150mL) were added to the reaction bottle, stirring and slowly adding DIEA (1158μL, 7mmol), 25°C reaction for 12 hours, removing the solvent under reduced pressure, adding 50mL EA and 50mL PE to beat, and precipitating white solid, repeating three times to obtain compound 14. MS (ESI): (M+H) + Calculated value 554.3, experimental value 554.4.

[0377] Compound 14 (1661mg, 3mmol), p-nitrophenyl chloroformate (1206mg, 6mmol) and THF (150mL) were added to the reaction bottle, stirring and adding Py (474μL, 6mmol) dropwise, 65°C reaction for 1 hour, removing the solvent under reduced pressure, and the residue was purified by silica gel chromatography (DCM / MeOH 98:2) to obtain compound 15 (1831mg, yield 85%). MS (ESI): (M+H) + Calculated value 719.8, experimental value 719.9. ​

[0378] Compound 15 (1436 mg, 2 mmol), exatecan mesylate 16 (1168.2 mg, 2.2 mmol) and N,N-dimethylformamide (100 mL) were added to a reaction bottle, stirred and slowly added DIEA (695 μL, 4 mmol), 25 °C for 20 hours, the solvent was removed under reduced pressure, and the residue was purified by silica gel chromatography (DCM / MeOH 95:5) to obtain compound 17 (1664 mg, yield 82%). MS (ESI): (M+H) + Calculated 1015.4, found 1015.1.

[0379] Compound 17 (1522 mg, 1.5 mmol), 40 mL THF (commercial) were added to a reaction bottle at 0 °C. Stirring and slowly adding DBU (228 mg, 1.5 mmol), the reaction was removed from the room temperature for 40 minutes after half an hour, until the TLC raw material disappeared. The reaction was concentrated under vacuum to remove THF, 15 mL DCM and 300 mL PE were added, a large amount of yellow-green solid was produced, and compound 18 solid was filtered out by Buchner funnel (1177 mg, yield 99%). MS (ESI): (M+H) + Calculated 793.3, found 793.6.

[0380] Compound 18 (1031 mg, 1.3 mmol), compound 8 (554 mg, 1.56 mmol), EDCI (498 mg, 2.6 mmol), HOBT (263 mg, 1.95 mmol) and N,N-dimethylformamide (20 mL) were added to a reaction bottle, 25 °C for 1 hour, the solvent was removed under reduced pressure, and the residue was purified by C18 (60% ACN / 0.05% formic acid in H2O) and freeze-dried to obtain compound 19 (1322 mg, yield 90%). MS (ESI): (M+H) + Calculated 1130.4, found 1130.7.

[0381] Compound 19 (903 mg, 0.8 mmol), compound 20 (702 mg, 1.2 mmol), tris(3-hydroxypropyltriazolylmethyl)amine (34.7 mg, 0.08 mmol) and cuprous bromide (11.4 mg, 0.08 mmol) were added to a reaction bottle, nitrogen was replaced three times, THF, DMF and H2O (3.5 mL:0.6 mL:0.4 mL) were added, 25 °C for 0.5 hours, and the residue was purified by C18 preparation (62% ACN / 0.05% formic acid in H2O) to obtain compound LD3 (1276 mg, yield 93%). MS (ESI): (M+H) + Calculated 1715.7, found 1716.2.1 H NMR (600 MHz, DMSO-d6) δ 10.71 (s, 1H), 10.08 (s, 1H), 8.48 (d, J = 6.7 Hz, 1H), 8.17 (d, J = 8.6 Hz, 3H), 8.09 (d, J = 8.8 Hz, 1H), 8.03 (d, J = 8.7 Hz, 2H), 7.89 (s, 1H), 7.81 (d, J = 10.8 Hz, 1H), 7.66 (d, J = 8.4 Hz, 2H), 7.42 - 7.36 (m, 3H), 6.61 (s, 1H), 6.02 (t, J = 6.8 Hz, 1H), 5.55 (s, 2H), 5.38 - 5.26 (m, 3H), 4.55 - 4.47 (m, 3H), 4.46 - 4.41 (m, 1H), 4.36 (q, J = 15.6 Hz, 2H), 4.28 (s, 2H), 3.80 (s, 5H), 3.62 - 3.55 (m, 36H), 3.52 - 3.47 (m, 9H), 3.44 - 3.36 (m, 2H), 3.30 - 2.29 (m, 1H), 3.19 - 3.09 (m, 1H), 2.42 (s, 3H), 2.28 - 2.19 (m, 1H), 2.17 (s, 3H), 2.17 - 2.10 (m, 1H), 2.04 - 1.89 (m, 2H), 1.42 (d, J = 7.0 Hz, 3H), 1.02 (d, J = 6.7 Hz, 3H), 0.98 (t, J = 7.2 Hz, 6H).

[0382] Example 11: Humanized antibody conjugation with LD1

[0383] Antibody reduction:

[0384] The antibody buffer system prepared in the laboratory was replaced with PBS7.0 / EDTA to prepare an antibody concentration of 10 mg / mL. The solution (1.0 mL) was placed in a 1.5 mL EP tube, 10 mM TCEP (Biolynx Technology Co., Ltd.) aqueous solution (16.7 μL) was added (2.5 equivalents relative to one molecule of antibody), and the antibody was incubated at 37°C water bath for 1 hour to reduce the interchain disulfide bond of the antibody.

[0385] Antibody conjugation with LD1:

[0386] To the above solution, 10 mM of DMSO solution of LD1 (33.3 μl; 5 equivalents relative to one molecule of antibody) was added at room temperature, mixed, and reacted at room temperature for 30 minutes to link LD1 to the free sulfhydryl group of the antibody. Next, 100 mM of aqueous N-ethylmaleimide (Promega) solution (10.0 μL; 15 equivalents relative to one molecule of antibody) was added, reacted at room temperature for 20 minutes, and the coupling reaction was terminated.

[0387] Purification of antibody-drug conjugate:

[0388] A 1 mL Mabselect SuRe pre-packed chromatography column was equilibrated with buffer A solution (20 mM sodium citrate, 500 mM NaCl, 0.02% polysorbate 20) on an AKTA system, antibody-drug conjugate was loaded using a loading loop, after re-equilibration with 10-12 mL of buffer A, impurities were washed using buffer B (20 mM sodium citrate), and finally eluted with buffer C (5 mM citric acid) and neutralized with 1 / 19 volume of sodium citrate neutralization solution to obtain the purified antibody-drug conjugate.

[0389] Determination of drug-antibody conjugation ratio (DAR) of antibody-drug conjugate:

[0390] Hydrophobic chromatography (HIC-HPLC) was used to determine the drug-antibody conjugation ratio (DAR) distribution and calculate the DAR value. Experimental instrument: high performance liquid chromatograph (e2695, Waters) Column: TSKgel Butyl-NPR, 4.6 mm*10 cm (item number 0042168 / TOSOH)

[0391] Experimental method:

[0392] Mobile phase: mobile phase A (1.0 M ammonium sulfate, 25 mM phosphate buffer), mobile phase B (25% isopropanol, 75% 25 mM phosphate buffer), and appropriate chromatographic conditions were selected. The experimental results were analyzed by area normalization method.

[0393] The prepared ADC and DAR value results are shown below:

[0394] Example 12: Anti-tumor experiment of antibody-drug conjugate on human pancreatic cancer HPAF-II mouse transplanted tumor

[0395] In this example, female NOD-SCID mice of appropriate age were inoculated with human pancreatic cancer HPAF-II cells, and when the tumor volume grew to about 100 mm 3When the tumor grew well, 35 animals were selected and divided into 7 groups according to the tumor volume. The animal grouping and dose schedule are shown in Table 11.

[0396] Table 11 Animal grouping and dose schedule

[0397] After grouping, the mice were dosed and weighed, and the data were recorded. The tumor growth was dynamically observed by measuring the tumor diameter at different times after dosing, and the tumor volume was calculated according to the following formula:

[0398] Tumor volume (mm 3 )= 1 / 2 x length (mm) x [short diameter (mm)] 2

[0399] Relative tumor volume: RTV = tumor volume (nth day) / tumor volume (0th day)

[0400] Relative tumor volume proliferation rate: T / C% = RTV (nth day) of the dosed group / RTV (nth day) of the solvent group x 100%

[0401] Tumor inhibition rate = (1-T / C) x 100%

[0402] On the 21st day after dosing, the mice were suffocated with carbon dioxide, and the tumor weight was measured after tumor peeling.

[0403] The experimental results were as follows: (1) all ADCs could significantly inhibit the growth of human pancreatic cancer HPAF-II transplanted tumors compared with the solvent group; (2) hu51-H1L2-LD1 and hu52-H2L1-LD1 had a significantly better effect on inhibiting the growth of HPAF-II transplanted tumors than TF011-LD1 and XB002-LD1. For details, see Table 12 and FIG. 5.

[0404] Table 12 Tumor parameters of HPAF-II transplanted tumor model after 21 days of dosing

[0405] Note: TTEST test, vs solvent group: ***P <0.0001, **P <0.01; vs XB002: ★★P <0.01, ★P <0.05, vs TF011: ◆◆P <0.01, ◆P <0.05.

[0406] Example 13: Anti-tumor experiment of antibody-drug conjugate on human pharyngeal squamous carcinoma FADU mouse transplanted tumor

[0407] In this example, female NU / NU mice of appropriate age were inoculated with human pharyngeal squamous carcinoma FADU cells, and when the tumor volume reached about 100 mm 3When the tumor grew well, 35 animals were selected, and the animals were divided into 7 groups according to the tumor volume, and the animal grouping and dose scheme are shown in Table 13.

[0408] Table 13 Animal grouping and dose scheme

[0409] After grouping, the mice were dosed and weighed, and the data were recorded. The tumor growth was dynamically observed by measuring the tumor diameter at different times after dosing, and the tumor volume was calculated according to the following formula:

[0410] Tumor volume (mm 3 ) = 1 / 2 x long diameter (mm) x [short diameter (mm)] 2

[0411] Relative tumor volume: RTV = tumor volume (nth day) / tumor volume (0th day)

[0412] Relative tumor volume proliferation rate: T / C% = RTV (nth day) of the dosed group / RTV (nth day) of the solvent group x 100

[0413] Tumor inhibition rate = (1-T / C) x 100%

[0414] On the 14th day after dosing, the mice were suffocated with carbon dioxide, and the tumor weight was measured after stripping.

[0415] Under the conditions of this experiment, (1) compared with the solvent group, all ADCs can significantly inhibit the growth of human pharyngeal squamous carcinoma FADU mouse transplanted tumor; (2) compared with Yangshen TF011-LD1 and XB002-LD1, hu51-H1L3-LD1 and hu52-H2L3-LD1 have the same effect on inhibiting the growth of FADU transplanted tumor as Yangshen, but the numerical value is slightly better than Yangshen TF011-LD1; the tumor weight inhibition rates of hu52-H2L1-LD1 and hu51-H1L2-LD1 are 96.4% and 89.3% respectively, which are significantly higher than Yangshen. For detailed information, see Table 14 and Figure 6.

[0416] Table 14 Tumor parameters of each group in FADU transplanted tumor model after 14 days of dosing

[0417] Note: TTEST test, vs solvent group, **P <0.01, ***P <0.001.

[0418] Example 14: Humanized antibody conjugated with LD2

[0419] Antibody reduction:

[0420] The antibody buffer system prepared in the laboratory was replaced with PBS 7.0 / EDTA to prepare an antibody solution of 10 mg / mL. The solution (1.0 mL) was placed in a 1.5 mL EP tube, and 10 mM TCEP (Sigma-Aldrich) aqueous solution (80.0 μL, 12 equivalents per molecule of antibody) was added thereto; the antibody interchain disulfide bond was reduced by incubation at 27.5°C for 3 hours.

[0421] Antibody conjugation with LD2:

[0422] To the above solution was added 20 mM LD2 DMSO solution (33.3 μL; 10 equivalents per molecule of antibody) at room temperature, mixed, and reacted at room temperature for 30 minutes to link LD2 to the antibody. Next, 100 mM N-ethylmaleimide (Sigma-Aldrich) aqueous solution (10.0 μL; 15 equivalents per molecule of antibody) was added, and further reacted at room temperature for 20 minutes to terminate the conjugation reaction.

[0423] Purification of antibody-drug conjugate, method same as Example 11.

[0424] Determination of DAR value of antibody-drug conjugate:

[0425] To the antibody-drug conjugate was added dithiothreitol at a final concentration of 20 mM, and the sample with the antibody-drug conjugate interchain disulfide bond cleaved was used for HPLC analysis. The HPLC system was Agilent Technologies 1260 Infinity HPLC, the chromatographic column was PLRP-S (5 μm particle size; 2.1 mm x 50 mm; Agilent Technologies), the column temperature was 80°C, the mobile phase A was 0.1% trifluoroacetic acid (TFA) aqueous solution, and the mobile phase B was 0.1% trifluoroacetic acid (TFA) acetonitrile solution. The sample size was 10 μL, and the gradient program was 0-3 minutes 27%-27%, 3-8 minutes 27%-35%, 8-25 minutes 35%-43%, 25-26 minutes 43%-95%, 26-31 minutes 95%-95%, 31-31.5 minutes 95%-27%, 31.5-40 minutes 27%-27%. The hydrophobicity of the light chain (light chain with one L-D, L1) and the heavy chain (heavy chain with one L-D, H1; heavy chain with two L-D, H2; heavy chain with three L-D, H3) to which the drug was linked increased with the number of L-Ds linked, relative to the light chain (L0) and the heavy chain (H0) to which no linker-drug compound (L-D) was linked, so they were eluted in the order of L0, L1, H0, H1, H2, H3. The DAR value was calculated based on the peak area at 280 nm.

[0426] ADC structure and DAR value are shown in Table 15.

[0427] Table 15 Summary of DAR values of antibody-drug conjugates

[0428] Example 15: Humanized antibody conjugation with LD3

[0429] Antibody reduction:

[0430] The antibody buffer system prepared in the laboratory was replaced with PBS7.0 / EDTA to prepare an antibody concentration of 10 mg / mL. The solution (1.0 mL) was placed in a 1.5 mL EP tube, and 10 mM TCEP (Biotang) aqueous solution was added (DAR6 (80.0 μL, 12 equivalents relative to one molecule of antibody); or DAR4 (40.0 μL, 6 equivalents relative to one molecule of antibody)); incubated at 27.5°C for 3 hours to reduce the interchain disulfide bond of the antibody.

[0431] Antibody conjugation with LD3:

[0432] To the above solution, 20 mM LD3 DMSO solution (33.3 μl; 10 equivalents relative to one molecule of antibody) was added at room temperature, mixed, and reacted at room temperature for 30 minutes to link LD3 to the antibody. Next, 100 mM N-ethylmaleimide (Biotang) aqueous solution (10.0 μL; 15 equivalents relative to one molecule of antibody) was added, and the reaction was allowed to proceed at room temperature for 20 minutes to terminate the conjugation reaction.

[0433] The antibody-drug conjugate was purified and the DAR value was determined as in Example 14. The ADC structure and DAR value are shown in Table 16.

[0434] Table 16 Summary of DAR values of antibody-drug conjugates

[0435] Example 16: Inhibition of human tumor cell growth in vitro by antibody-drug conjugates

[0436] The hu51-H1L2 series of antibodies conjugated with different toxin ADC molecules were selected to evaluate their in vitro growth inhibition activity on human cervical cancer cells CaSki and human pancreatic cancer cells HPAF-II. The target cells were collected and resuspended as a single cell suspension, and the cell viability was identified by trypan blue staining and cell counting; the cell density was adjusted to 1×10 5Cells / mL; 100 μL per well was added to a 96-well black flat-bottom cell culture plate; 20 μL of diluted ADC test article was added to the 96-well black flat-bottom cell culture plate seeded with cells; incubated in a cell culture incubator (37°C, 5% CO2) for about 168 hours; added resazurin sodium solution (0.03%), 20 μL per well; reacted at 37°C for 3-4 hours, read fluorescence value at 550 nm by microplate reader, plotted by Prism plotting software, fitted half-inhibitory concentration IC50 of test article 50 .

[0437] In the in vitro experiment, hu51-H1L2-LD2, hu51-H1L2-6-LD2, hu51-H1L2-6-LD3 and hu51-H1L2-4-LD3 all have good cell growth inhibition activity. The results are shown in Table 17.

[0438] Table 17 In vitro growth inhibition activity of antibody-drug conjugates

[0439] Example 17: Anti-tumor experiment of antibody-drug conjugates on human cervical cancer CaSki mouse transplanted tumor

[0440] In this example, female NOD-SCID mice of appropriate age were inoculated with human cervical cancer Caski cells, and when the tumor volume grew to about 100 mm 3 , 20 animals with good tumor growth were selected, and the animals were divided into 4 groups according to the tumor volume. The animal grouping and dose regimen are shown in Table 18.

[0441] Table 18 Animal grouping and dose regimen

[0442] After grouping, the mice were dosed and weighed, and the data were recorded. The growth of the tumor was dynamically observed by measuring the tumor diameter at different times after dosing, and the tumor volume was calculated according to the following formula:

[0443] Tumor volume (mm 3 ) = 1 / 2 x length (mm) x [short diameter (mm)] 2

[0444] Relative tumor volume: RTV = tumor volume (nth day) / tumor volume (0th day)

[0445] Relative tumor volume proliferation rate: T / C% = RTV of dosed group (nth day) / RTV of solvent group (nth day) x 100

[0446] Tumor inhibition rate = (1-T / C) x 100%

[0447] On day 28 after administration, the mice were suffocated with carbon dioxide, and the tumor weight was measured after tumor peeling.

[0448] The results showed that the ADCs (hu51-H1L2-LD2, hu51-H1L2-6-LD2 and hu51-H1L2-6-LD3) of the application could inhibit the growth of human cervical cancer CaSki transplanted tumors, and hu51-H1L2-LD2 and hu51-H1L2-6-LD3 had stronger abilities to inhibit the growth of human cervical cancer CaSki transplanted tumors. For details, see Table 19 and FIG. 7.

[0449] Table 19: Tumor parameters of each group in the CaSki transplanted tumor model after 28 days of administration

[0450] Note: ***p<0.001, **p<0.01, p<0.05 VS vehicle group

[0451] Example 18: Anti-tumor experiment of antibody-drug conjugate on human cervical cancer HT-3 mouse transplanted tumor

[0452] In this example, female NOD-SCID mice of the appropriate age were inoculated with human cervical cancer HT-3 cells, and when the tumor volume reached about 115 mm 3 When the tumor volume reached about 115 mm 3 , 25 animals with good tumor growth were selected, and the animals were divided into 5 groups according to the tumor volume. The animal grouping and dose scheme are shown in Table 20. IgG-LD3 was prepared in the same way as in Example 15 (IgG is a monoclonal antibody against gp120 (HIV viral envelope surface glycoprotein), which is used as a negative reference in this experiment, and the sequence is from patent US11597759B2), wherein 80.0 μL of 10 mM TCEP (Tris (2-carboxyethyl) phosphine hydrochloride) aqueous solution (12 equivalents relative to one molecule of antibody) was added; 23.3 μl of 20 mM LD3 DMSO solution (7 equivalents relative to one molecule of antibody) was added; and 10.0 μL of 100 mM N-ethylmaleimide (Tris (2-carboxyethyl) phosphine hydrochloride) aqueous solution (15 equivalents relative to one molecule of antibody) was added. The ADC purification and DAR detection methods are the same as in Examples 11 and 15.

[0453] Table 20: Animal grouping and dose scheme

[0454] After grouping, the mice were administered and weighed, and the data was recorded. The tumor growth was dynamically observed by measuring the tumor diameters at different times after administration, and the tumor volume was calculated according to the following formula:

[0455] Tumor volume (mm 3 ) = 1 / 2 x long diameter (mm) x [short diameter (mm)] 2

[0456] Relative tumor volume: RTV = tumor volume (nth day) / tumor volume (0th day)

[0457] Relative tumor volume proliferation rate: T / C% = RTV (nth day) of administration group / RTV (nth day) of solvent group x 100

[0458] Tumor inhibition rate = (1-T / C) x 100%

[0459] On the 24th day after administration, the tumor weight was measured after the mice were suffocated with carbon dioxide.

[0460] The experimental results show that, under the test conditions, (1) compared with the solvent group, the tumor inhibition effect of TF011-LD1 and IgG-LD3 groups is not good, and the TF011-LD1 group may be caused by the HT-3 tumor being insensitive to TF011-LD1 or its toxin molecules, and the IgG-LD3 group is caused by the lack of targeting of the antibody; (2) compared with the positive TF011-LD1, the ADC of the present application significantly outperforms the positive TF011-LD1 in inhibiting the growth of HT-3 transplanted tumors. For detailed information, see Table 21 and FIG. 8.

[0461] Table 21 Tumor parameters of each group in HT-3 transplanted tumor model after 24 days of administration

[0462] Note: *** p < 0.001, ** p < 0.01, p < 0.05 VS solvent group; # p < 0.05 ## p < 0.01 VS TF011-LD1

Claims

1. An anti-tissue factor (TF) antibody or antigen-binding fragment thereof, the antibody or antigen-binding fragment comprising a heavy chain variable region comprising three complementarity determining regions HCDR1, HCDR2, and HCDR3, and a light chain variable region comprising three complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein: (1) the amino acid sequence of HCDR1 is SEQ ID NO. 5 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 5, the amino acid sequence of HCDR2 is SEQ ID NO. 6 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 6, the amino acid sequence of HCDR3 is SEQ ID NO. 7 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 7, the amino acid sequence of LCDR1 is SEQ ID NO. 14 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 14, the amino acid sequence of LCDR2 is SEQ ID NO. 15 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 15, and the amino acid sequence of LCDR3 is SEQ ID NO. 16 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 16; or (2) the amino acid sequence of HCDR1 is SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, the amino acid sequence of HCDR2 is SEQ ID NO. 9 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 9, the amino acid sequence of HCDR3 is SEQ ID NO. 10 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 10, the amino acid sequence of LCDR1 is SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17, the amino acid sequence of LCDR2 is SEQ ID NO. 18 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 18, and the amino acid sequence of LCDR3 is SEQ ID NO. 19 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO.

19. ​ (2) the amino acid sequence of HCDR1 is set forth in SEQ ID NO. 20 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, the amino acid sequence of HCDR2 is set forth in SEQ ID NO. 21 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, the amino acid sequence of HCDR3 is set forth in SEQ ID NO. 22 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, the amino acid sequence of LCDR1 is set forth in SEQ ID NO. 27 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, the amino acid sequence of LCDR2 is set forth in SEQ ID NO. 28 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, and the amino acid sequence of LCDR3 is set forth in SEQ ID NO. 29 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto; wherein the HCDR and LCDR sequences are defined according to Kabat.

2. The antibody or antigen-binding fragment thereof of claim 1, wherein the amino acid sequence of the heavy chain variable region is set forth in any one of SEQ ID NOs. 1-4, 18, and 19 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto.

3. The antibody or antigen-binding fragment thereof of claim 1 or 2, wherein the amino acid sequence of the light chain variable region is set forth in any one of SEQ ID NOs. 11-13 and 24-26 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto.

4. The antibody or antigen-binding fragment thereof of any one of claims 1-3, wherein: (1) the amino acid sequence of the heavy chain variable region is set forth in any one of SEQ ID NOs. 1-4 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, and the amino acid sequence of the light chain variable region is set forth in any one of SEQ ID NOs. 11-13 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto; or (2) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18 or 19, or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 18 or 19, and the amino acid sequence of the light chain variable region is set forth in any one of SEQ ID NO. 24-26, or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in any one of SEQ ID NO. 24-26.

5. The antibody or antigen-binding fragment thereof of claim 4, wherein: (1) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1, or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 1, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11, or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 11; or (2) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1, or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 1, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12, or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 12; or (3) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1, or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 1, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13, or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 13; or (4) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2, or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 2, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11, or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 11; or (5) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 2, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 12; or (6) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 2, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 13; or (7) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 3 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 3, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11; or (8) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 3 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 3, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 12; or (9) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 3 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 3, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 13; or (10) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 4 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 4, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11; or (11) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 4 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 4, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 12; or (12) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 4 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 4, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 13; or (13) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 18, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 24 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 24; or (14) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 18, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 25 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 25; or (15) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 18, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 26 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 26; or (16) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 19 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 19, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 24 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 24; or (17) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 19 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 19, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 25 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 25; or (18) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 19 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 19, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 26 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 26; (19) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 36 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 36, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 37 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 37; or (20) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 38 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 38, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 39 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO.

39.

6. The antibody or antigen-binding fragment thereof of any one of claims 1-5, further comprising a heavy chain constant region, wherein: the amino acid sequence of the heavy chain constant region is set forth in any one of SEQ ID NOs. 8-10 and 23 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to any one of SEQ ID NOs. 8-10 and 23.

7. The antibody or antigen-binding fragment thereof of any one of claims 1-6, further comprising a light chain constant region, wherein: the amino acid sequence of the light chain constant region is the sequence set forth in any one of SEQ ID NO. 17, 30-31 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in any one of SEQ ID NO. 17, 30-31.

8. The antibody or antigen-binding fragment thereof of any one of claims 1-7, further comprising a heavy chain constant region and a light chain constant region, wherein: (1) the amino acid sequence of the heavy chain variable region is the sequence set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 1, the amino acid sequence of the light chain variable region is the sequence set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is the sequence set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is the sequence set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 17; (2) the amino acid sequence of the heavy chain variable region is the sequence set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 1, the amino acid sequence of the light chain variable region is the sequence set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is the sequence set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is the sequence set forth in SEQ ID NO. 31 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 31; (3) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 9 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 9, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; (4) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 9 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 9, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 31 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 31; (5) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 10 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 10, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; (6) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 10 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 10, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 31 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 31; (7) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 18, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 24 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 24, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 30; (8) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 18, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 25 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 25, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 30; (9) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 18, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 26 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 26, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 30; (10) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 19 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 19, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 24 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 24, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 30; (11) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 19 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 19, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 25 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 25, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 30; (12) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 19 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 26 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 30 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto; (13) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto; (14) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto; (15) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 2, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; (16) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 2, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; (17) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 2 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 2, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 13, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to SEQ ID NO. 17; (18) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 3 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 3, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 17; (19) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 3 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 3, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 17; (20) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 3 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 3, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 13, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical to SEQ ID NO. 17; (21) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 4 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto; (22) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 4 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto; (23) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 4 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 13 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17 or a sequence at least 80%, 85%, 90%, 95%, 98%, or 99% identical thereto; or (24) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 10 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO. 10, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 31 or a sequence having at least 80%, 85%, 90%, 95%, 98%, or 99% identity to the sequence set forth in SEQ ID NO.

31.

9. The antibody or antigen-binding fragment thereof according to any one of claims 1-8, wherein: the antigen-binding fragment is an Fv, scFv, Fab, Fab', or (Fab')2; and / or the antibody or antigen-binding fragment thereof is a murine, chimeric, or humanized antibody or antigen-binding fragment, preferably a humanized antibody or antigen-binding fragment; and / or the antibody or antigen-binding fragment thereof binds to human TF, preferably the antibody or antigen-binding fragment thereof binds to human and cynomolgus TF, but not to rat or mouse TF; and / or the antibody or antigen-binding fragment thereof has ADCC activity; and / or the antibody or antigen-binding fragment thereof has CDC activity; and / or the antibody or antigen-binding fragment thereof has ADCP activity; and / or the antibody is a full-length antibody; and / or the antibody is a monoclonal antibody; and / or the antibody is an IgG, IgM, IgD, IgA, or IgE type antibody, preferably an IgG type antibody, more preferably an IgGl type antibody; and / or the antibody comprises a light chain constant region of the kappa or lambda subtype; and / or the antibody comprises a human IgGl type heavy chain constant region and a human kappa subtype light chain constant region; and / or the antibody comprises a wild-type Fc region; and / or the antibody or antigen-binding fragment thereof is engineered to have enhanced ADCC, CDC, and / or ADCP activity, preferably the antibody comprises an engineered Fc region that confers to the antibody enhanced ADCC, CDC, and / or ADCP effector function; and / or the antibody or antigen-binding fragment thereof is engineered to have reduced or ablated ADCC, CDC, and / or ADCP activity, preferably the antibody comprises an engineered Fc region that confers to the antibody reduced or ablated ADCC, CDC, and / or ADCP effector function.

10. An anti-tissue factor (TF) antibody, comprising a heavy chain variable region, a light chain variable region, a heavy chain constant region, and a light chain constant region, wherein: (1) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17; (2) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 31; (3) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 9, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17; (4) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 9, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 31; (5) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 10, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 17; (6) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 10, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO. 31; (7) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 18, the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 24, the amino acid sequence of the heavy chain constant region is set forth in SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is set forth in SEQ ID NO.

30. ​ ​ ​ ​ ​ ​ ​ (8) the amino acid sequence of the heavy chain variable region is represented by SEQ ID NO. 18, the amino acid sequence of the light chain variable region is represented by SEQ ID NO. 25, the amino acid sequence of the heavy chain constant region is represented by SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is represented by SEQ ID NO. 30; (9) the amino acid sequence of the heavy chain variable region is represented by SEQ ID NO. 18, the amino acid sequence of the light chain variable region is represented by SEQ ID NO. 26, the amino acid sequence of the heavy chain constant region is represented by SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is represented by SEQ ID NO. 30; (10) the amino acid sequence of the heavy chain variable region is represented by SEQ ID NO. 19, the amino acid sequence of the light chain variable region is represented by SEQ ID NO. 24, the amino acid sequence of the heavy chain constant region is represented by SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is represented by SEQ ID NO. 30; (11) the amino acid sequence of the heavy chain variable region is represented by SEQ ID NO. 19, the amino acid sequence of the light chain variable region is represented by SEQ ID NO. 25, the amino acid sequence of the heavy chain constant region is represented by SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is represented by SEQ ID NO. 30; (12) the amino acid sequence of the heavy chain variable region is represented by SEQ ID NO. 19, the amino acid sequence of the light chain variable region is represented by SEQ ID NO. 26, the amino acid sequence of the heavy chain constant region is represented by SEQ ID NO. 23, and the amino acid sequence of the light chain constant region is represented by SEQ ID NO. 30; (13) the amino acid sequence of the heavy chain variable region is represented by SEQ ID NO. 1, the amino acid sequence of the light chain variable region is represented by SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is represented by SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is represented by SEQ ID NO. 17; (14) the amino acid sequence of the heavy chain variable region is represented by SEQ ID NO. 1, the amino acid sequence of the light chain variable region is represented by SEQ ID NO. 13, the amino acid sequence of the heavy chain constant region is represented by SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is represented by SEQ ID NO. 17; (15) the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO. 2, the amino acid sequence of the light chain variable region is shown in SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is shown in SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is shown in SEQ ID NO. 17; (16) the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO. 2, the amino acid sequence of the light chain variable region is shown in SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is shown in SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is shown in SEQ ID NO. 17; (17) the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO. 2, the amino acid sequence of the light chain variable region is shown in SEQ ID NO. 13, the amino acid sequence of the heavy chain constant region is shown in SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is shown in SEQ ID NO. 17; (18) the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO. 3, the amino acid sequence of the light chain variable region is shown in SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is shown in SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is shown in SEQ ID NO. 17; (19) the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO. 3, the amino acid sequence of the light chain variable region is shown in SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is shown in SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is shown in SEQ ID NO. 17; (20) the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO. 3, the amino acid sequence of the light chain variable region is shown in SEQ ID NO. 13, the amino acid sequence of the heavy chain constant region is shown in SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is shown in SEQ ID NO. 17; (21) the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO. 4, the amino acid sequence of the light chain variable region is shown in SEQ ID NO. 11, the amino acid sequence of the heavy chain constant region is shown in SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is shown in SEQ ID NO. 17; (22) the amino acid sequence of the heavy chain variable region is depicted in SEQ ID NO. 4, the amino acid sequence of the light chain variable region is depicted in SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is depicted in SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is depicted in SEQ ID NO. 17; (23) the amino acid sequence of the heavy chain variable region is depicted in SEQ ID NO. 4, the amino acid sequence of the light chain variable region is depicted in SEQ ID NO. 13, the amino acid sequence of the heavy chain constant region is depicted in SEQ ID NO. 8, and the amino acid sequence of the light chain constant region is depicted in SEQ ID NO. 17; or (24) the amino acid sequence of the heavy chain variable region is depicted in SEQ ID NO. 1, the amino acid sequence of the light chain variable region is depicted in SEQ ID NO. 12, the amino acid sequence of the heavy chain constant region is depicted in SEQ ID NO. 10, and the amino acid sequence of the light chain constant region is depicted in SEQ ID NO.

31.

11. An isolated nucleic acid molecule encoding the anti-TF antibody or antigen binding fragment thereof according to any one of claims 1-10, or a combination of nucleic acid molecules encoding the heavy chain and light chain of the anti-TF antibody according to any one of claims 1-10, respectively.

12. A vector (e.g., an expression vector) comprising the nucleic acid molecule or combination of nucleic acid molecules of claim 11.

13. A cell comprising the nucleic acid molecule or combination of nucleic acid molecules of claim 11 or the vector of claim 12.

14. An antibody-drug conjugate having the structure depicted in Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled form thereof: A-(L-D)d (I) wherein: A is the anti-TF antibody or antigen binding fragment thereof according to any one of claims 1-10; D is a pharmaceutically active molecule, preferably a molecule having a disease- treating activity, more preferably a molecule having an anticancer activity, most preferably a small molecule having an anticancer activity; L is a linker moiety, one end of which is covalently linked to A and one end of which is covalently linked to the pharmaceutically active molecule D; and d is an integer or decimal number in the range of 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12).

15. The antibody-drug conjugate of claim 14, or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled form thereof, wherein the structure of -L- is -L1-L2-L3-L4-, L1 is a covalent linking unit covalently linked to A, L2 is an extension unit, L3 is a peptide residue consisting of 2-8 amino acids, and L4 is a bond or a self-cleaving fragment. ​ ​ ​ ​ ​ ​ 16. The antibody-drug conjugate or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled material thereof of claim 15, wherein, L2is selected from -L 2a -, -L 2b -, -C(O)-, -NH-, -SO2-, -CH2-; preferably L2is selected from -L 2a -, -L 2a -, -C(O)-, -L 2a - L 2b -, -C(O)-, -L 2a -, -NH-C(O)-, -L 2a - L 2b -, -NH-C(O)-, -L 2a - L 2b -, -C(O)-NH-, -L 2a -, -C(O)-NH-L 2b -, -C(O)-, -L 2a -, -NH-C(O)-L 2b -, -C(O)-, -L 2a -, -C(O)-NH-L 2b -, -C(O)-NH- or -L 2a -, -NH-C(O)-L 2b -, -NH-C(O)-, wherein L 2a , L 2b each independently is selected from -C1-C8alkylene-, -C1-C8alkylene-C3-C8cycloalkylene-, -C2-C8alkynylene-, -C6-C 14 arylene-, -C6-C 14 arylene-C1-C8alkylene-, -5-10 membered heteroarylene-, -5-10 membered heteroarylene-C1-C8alkylene-, straight or branched heteroalkylene of 1-50 carbon atoms, straight or branched heteroalkylene-3-8 membered heterocyclyl of 1-50 carbon atoms, each of said alkylene, cycloalkylene, arylene, heteroalkylene, heteroarylene, heterocyclylene is optionally substituted with one or more substituents independently selected from halogen, C1-C6alkyl, heteroalkyl of 1-6 carbon atoms (such as C1-C6alkoxy), hydroxyl, amino, carboxyl, or C3-C8cycloalkyl, said heteroalkylene, heterocyclylene, heteroalkyl containing 1-12 heteroatoms, said heteroatoms of said heteroalkylene, heterocyclylene, heteroarylene, heteroalkyl selected from one or more of N, O, or S; and / or L3 is a peptide residue consisting of 2-8 natural or non-natural amino acids, which are optionally further substituted with one or more substituents selected from C1-C6 alkyl, heteroalkyl of 2-6 atoms, C1-C6 alkoxy, hydroxyl, amino, carboxyl, or C3-C8 cycloalkyl; preferably, the amino acids are selected from the group consisting of: phenylalanine, isoleucine, leucine, tryptophan, valine, methionine, tyrosine, alanine, threonine, histidine, serine, glutamine, arginine, lysine, asparagine, glutamic acid, proline, citrulline, aspartic acid, and glycine; and / or L4is selected from: -NCH2-, wherein the *position is attached to L3, wherein R1is selected from the group consisting of hydrogen and wherein R2 is a hydrophilic segment, preferably the hydrophilic segment comprises a polyethylene glycol, a poly natural or non-natural amino acid group, a monosaccharide, an oligosaccharide, or a polysaccharide, or a combination thereof; more preferably the hydrophilic segment comprises a straight chain or branched heteroalkyl of 4-50 -OCH2CH2- structural units, a peptide chain comprising 4-50 proteinogenic or non-proteinogenic amino acids, or a straight chain or branched heteroalkyl comprising a monosaccharide, an oligosaccharide, or a polysaccharide.

17. The antibody-drug conjugate or pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled material thereof of any one of claims 15-16, wherein L1is selected from wherein * indicates attachment to A; and / or L2is selected from -L 2a - and -L 2a - and -L 2a - and -L 2b - and -L 2a - and -L 2b each independently is selected from -C1-C6alkylene-, -C1-C3alkylene-C3-C6cycloalkylene-, -ethynylene-C1-C6alkylene-, -(CH2CH2O)n-, n - and -(CH2CH2O)n- n C1-C6alkylene-, -C1-C3alkylene-C3-C6cycloalkylene-, -ethynylene-C1-C6alkylene-, -(CH2CH2O)n-, 1-3 C1-C6alkylene-, -C1-C3alkylene-C3-C6cycloalkylene-, -ethynylene-C1-C6alkylene-, -(CH2CH2O)n-, 1-3 C1-C6alkylene-, -C1-C3alkylene-C3-C6cycloalkylene-, -ethynylene-C1-C6alkylene-, -(CH2CH2O)n-, n C1-C6alkylene-, -C1-C3alkylene-C3-C6cycloalkylene-, -ethynylene-C1-C6alkylene-, -(CH2CH2O)n-, 1-3 C1-C6alkylene-, -C1-C3alkylene-C3-C6cycloalkylene-, -ethynylene-C1-C6alkylene-, -(CH2CH2O)n-, 1-3 C1-C6alkylene-, -C1-C3alkylene-C3-C6cycloalkylene-, -ethynylene-C1-C6alkylene-, -(CH2CH2O)n-, 1-3 C1-C6alkylene-, -C1-C3alkylene-C3-C6cycloalkylene-, -ethynylene-C1-C6alkylene-, -(CH2CH2O)n-, 1-3 C1-C6alkylene-, -C1-C3alkylene-C3-C6cycloalkylene-, -ethynylene-C1-C6alkylene-, -(CH2CH2O)n-, wherein n is an integer selected from 1 to 12 (such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, preferably an integer from 3 L3 is a peptide residue consisting of 2-8 natural or non-natural amino acids, which are optionally further substituted with one or more substituents selected from C1-C6 alkyl, heteroalkyl of 2-6 atoms, C1-C6 alkoxy, hydroxyl, amino, carboxyl, or C3-C8 cycloalkyl; preferably, the amino acids are selected from the group consisting of: phenylalanine, isoleucine, leucine, tryptophan, valine, methionine, tyrosine, alanine, threonine, histidine, serine, glutamine, arginine, lysine, asparagine, glutamic acid, proline, citrulline, aspartic acid, and glycine; and / or L4is selected from: a bond, -NCH2-, wherein R1is selected from hydrogen and R2is selected from: wherein r, s, t, u are each independently an integer selected from 1-50 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50).

18. The antibody-drug conjugate or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled form thereof of any one of claims 15-17, wherein, L2is selected from: (CH2CH20) n CH2C(O)-, -(CH2CH20) n CH2CH2-, wherein the * position is attached to L1, and n is as defined in claim 17.

19. The antibody-drug conjugate or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled form thereof of any one of claims 15-18, wherein, L3 has a structure selected from the group consisting of: wherein the * position is attached to L2.

20. The antibody-drug conjugate or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled form thereof of any one of claims 15-19, wherein, L4 is selected from: -NCH2-, or L4 is selected from: wherein r is an integer selected from 1-50 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50).

21. The antibody-drug conjugate or pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled material thereof of any one of claims 15-20, wherein L has a structure selected from the group consisting of: wherein * indicates attachment to A, preferably wherein R2is r is an integer from 8 to 24, for example 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24.

22. The antibody-drug conjugate or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled thereof of any one of claims 14-21, wherein D is a molecule having anticancer activity selected from a cytotoxic drug, an immunopotentiator, and a radioisotope; Preferably, wherein the cytotoxic drug is selected from a tubulin inhibitor, a DNA topoisomerase inhibitor, a DNA damaging agent, an antimetabolite, or an antitumor antibiotic; Preferably, wherein the tubulin inhibitor is selected from an auristatin derivative (e.g. MMAE (Monomethyl auristatin E), MMAF (Monomethyl auristatin F)) or a maytansinoid derivative (e.g. DM1, DM4, Ansamitocin, Mertansine or dolastatin and derivatives thereof); Preferably, wherein the DNA topoisomerase inhibitor is selected from a camptothecin analogue or a DNA topoisomerase I inhibitor and derivatives thereof, for example, DXD, SN38, irinotecan, irinotecan hydrochloride, camptothecin, 9-amino camptothecin, 9-nitrocamptothecin, 10-hydroxycamptothecin, 9-chloro-10-hydroxycamptothecin, 22-hydroxy camptothecin, topotecan, lurtotecan, belotecan, exatecan, silatecan, 6,8-dibromo-2-methyl-3-[2-(D-xylopyranosylamino)phenyl]-4(3H)-quinazolinone, 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-(2E)-2-propenamide, 2-cyano-3-(3,4-dihydroxyphenyl)-N-(3-hydroxyphenylpropyl)-(E)-2-propenamide, 12- -D-xylopyranosyl-12,13-dihydro-2,10-dihydroxy-6-[[2-hydroxy-1-(hydroxymethyl)ethyl]amino]-5H-indolo[2,3-a]pyrrolo[3,4-c]carbazol-5,7(6H)-dione, N-[2-(dimethylamino)ethyl]-4-acridinecarboxamide dihydrochloride, N-[2-(dimethylamino)ethyl]-4-acridinecarboxamide, and derivatives thereof; Preferably, wherein the DNA damaging agent is selected from a calicheamicin, a duocarmycin, an anthramycin derivative PBD (pyrrolobenzodiazepine); Preferably, wherein the immune enhancer is selected from levamisole, pidotimod, imiquimod, isoprinosine, poliinosinic acid or poliinosinic acid; and / or the antimetabolite is selected from methotrexate, 6-mercaptopurine or 5-fluorouracil; and / or the antitumor antibiotic is selected from polypeptide antibiotics (e.g. actinomycin D or bleomycin) or anthraquinones (e.g. doxorubicin or mitoxantrone); Preferably, wherein the radioisotope is selected from 211 At, 131 I, 125 I, 90 Y, 186 Re, 188 Re, 153 Sm, 212 Bi, 32 P, 60 Co or 177 Lu.

23. The antibody-drug conjugate or pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled material thereof of any one of claims 14-21, wherein D has the following structure: wherein X is selected from a bond or wherein R3, R4are each independently selected from H, C 1-3 alkyl (preferably methyl, ethyl, n-propyl or i-propyl), 3-6 membered cycloalkyl (preferably cyclopropanyl, cyclobutanyl, cyclopentanoyl or cyclohexanoyl), or R3and R4together with the carbon atom to which they are attached form a 3-6 membered cycloalkyl (preferably cyclopropanyl, cyclobutanoyl, cyclopentanoyl or cyclohexanoyl), wherein the * position is attached to L (when the structure of -L- is -L1-L2-L3-L4-, to L4).

24. The antibody-drug conjugate or pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled material thereof of any one of claims 14-23, wherein D has a structure selected from the group consisting of:

25. The antibody-drug conjugate or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled material thereof of any one of claims 14-24, having a structure selected from the group consisting of: wherein r is an integer selected from 1-50, preferably 8-24, preferably 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24; and d is an integer or a decimal number selected from 1 to 12, preferably 4, 6 or 8.

26. The antibody-drug conjugate or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled material thereof of any one of claims 14-25, having a structure selected from the group consisting of: wherein A is an anti-TF antibody comprising a heavy chain variable region of the amino acid sequence set forth in SEQ ID NO. 1, a light chain variable region of the amino acid sequence set forth in SEQ ID NO. 12, a heavy chain constant region of the amino acid sequence set forth in SEQ ID NO. 8, and a light chain constant region of the amino acid sequence set forth in SEQ ID NO. 17, or an anti-TF antibody comprising a heavy chain variable region of the amino acid sequence set forth in SEQ ID NO. 1, a light chain variable region of the amino acid sequence set forth in SEQ ID NO. 13, a heavy chain constant region of the amino acid sequence set forth in SEQ ID NO. 8, and a light chain constant region of the amino acid sequence set forth in SEQ ID NO. 17, or an anti-TF antibody comprising a heavy chain variable region of the amino acid sequence set forth in SEQ ID NO. 19, a light chain variable region of the amino acid sequence set forth in SEQ ID NO. 24, a heavy chain constant region of the amino acid sequence set forth in SEQ ID NO. 23, and a light chain constant region of the amino acid sequence set forth in SEQ ID NO. 30, or an anti-TF antibody comprising a heavy chain variable region of the amino acid sequence set forth in SEQ ID NO. 19, a light chain variable region of the amino acid sequence set forth in SEQ ID NO. 26, a heavy chain constant region of the amino acid sequence set forth in SEQ ID NO. 23, and a light chain constant region of the amino acid sequence set forth in SEQ ID NO. 30, and d is an integer or decimal number selected from 1 to 12, preferably 4, 6, or 8; or wherein A is an anti-TF antibody comprising a heavy chain variable region of an amino acid sequence as set forth in SEQ ID NO. 1, a light chain variable region of an amino acid sequence as set forth in SEQ ID NO. 12, a heavy chain constant region of an amino acid sequence as set forth in SEQ ID NO. 8, and a light chain constant region of an amino acid sequence as set forth in SEQ ID NO. 17, or an anti-TF antibody comprising a heavy chain variable region of an amino acid sequence as set forth in SEQ ID NO. 1, a light chain variable region of an amino acid sequence as set forth in SEQ ID NO. 12, a heavy chain constant region of an amino acid sequence as set forth in SEQ ID NO. 9, and a light chain constant region of an amino acid sequence as set forth in SEQ ID NO. 17, and d is an integer or decimal number selected from 1 to 12, preferably 4, 6, or 8; or wherein A is an anti-TF antibody comprising a heavy chain variable region of an amino acid sequence as set forth in SEQ ID NO. 1, a light chain variable region of an amino acid sequence as set forth in SEQ ID NO. 12, a heavy chain constant region of an amino acid sequence as set forth in SEQ ID NO. 9 and a light chain constant region of an amino acid sequence as set forth in SEQ ID NO. 17, or an anti-TF antibody comprising a heavy chain variable region of an amino acid sequence as set forth in SEQ ID NO. 1, a light chain variable region of an amino acid sequence as set forth in SEQ ID NO. 12, a heavy chain constant region of an amino acid sequence as set forth in SEQ ID NO. 10 and a light chain constant region of an amino acid sequence as set forth in SEQ ID NO. 31, and d is an integer or a decimal number selected from 1 to 12, preferably 4, 6 or 8.

27. A pharmaceutical composition comprising the anti-Tissue Factor (TF) antibody or antigen binding fragment thereof of any one of claims 1-10 or the antibody-drug conjugate of any one of claims 14-26, or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer or isotopically-labeled material thereof, and a pharmaceutically acceptable carrier or excipient.

28. The pharmaceutical composition of claim 27 for use in the treatment and / or prevention of a TF-mediated disease or disorder.

29. The pharmaceutical composition of claim 28, wherein the disease or disorder is cancer, preferably a TF-positive cancer.

30. The pharmaceutical composition of claim 29, wherein the cancer is a solid tumor, for example, selected from cervical cancer, pancreatic cancer, head and neck cancer (e.g., pharyngeal squamous carcinoma), endometrial cancer, esophageal cancer, prostate cancer, non-small cell lung cancer and ovarian cancer.

31. The pharmaceutical composition of any one of claims 27-30, further comprising one or more additional therapeutic agents.

32. Use of the anti-tissue factor (TF) antibody or antigen-binding fragment thereof of any one of claims 1-10, or the antibody-drug conjugate or pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled material thereof of any one of claims 14-26, or the pharmaceutical composition of claim 27, for the manufacture of a medicament for treating and / or preventing a TF-mediated disease or disorder.

33. The use of claim 32, wherein the disease or disorder is cancer, preferably a TF-positive cancer.

34. The use of claim 33, wherein the cancer is a solid tumor, for example, selected from cervical cancer, pancreatic cancer, head and neck cancer (e.g., pharyngeal squamous carcinoma), endometrial cancer, esophageal cancer, prostate cancer, non-small cell lung cancer, and ovarian cancer.

35. A method of treating and / or preventing a TF-mediated disease or disorder in an individual, comprising administering to an individual in need thereof an effective amount of the anti-tissue factor (TF) antibody or antigen-binding fragment thereof of any one of claims 1-10, or the antibody-drug conjugate or pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled material thereof of any one of claims 14-26, or the pharmaceutical composition of claim 27.

36. The method of claim 35, wherein the disease or disorder is cancer, preferably a TF-positive cancer.

37. The method of claim 36, wherein the cancer is a solid tumor, for example, selected from cervical cancer, pancreatic cancer, head and neck cancer (e.g., pharyngeal squamous carcinoma), endometrial cancer, esophageal cancer, prostate cancer, non-small cell lung cancer, and ovarian cancer.

38. A pharmaceutical combination comprising the anti-tissue factor (TF) antibody or antigen-binding fragment thereof of any one of claims 1-10, or the antibody-drug conjugate or pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled material thereof of any one of claims 14-26, or the pharmaceutical composition of claim 27, and one or more additional therapeutic agents.

39. A kit comprising the anti-tissue factor (TF) antibody or antigen-binding fragment thereof of any one of claims 1-10, or the antibody-drug conjugate or pharmaceutically acceptable salt, hydrate, solvate, tautomer, stereoisomer, or isotopically-labeled material thereof of any one of claims 14-26, or the pharmaceutical composition of claim 27.

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