Anti-TL1a antibody, preparation method therefor and use thereof

Abstract

Disclosed in the present invention are an anti-TL1A antibody, a preparation method therefor and the use thereof. The antibody comprises a heavy chain variable region and a light chain variable region, and has one or more of the following technical features: the heavy chain variable region comprises HCDR1 having an amino acid sequence as shown in SEQ ID NO: 10 or 18; the heavy chain variable region comprises HCDR2 having an amino acid sequence as shown in SEQ ID NO: 11, 19, 39, 40 or 41; the heavy chain variable region comprises HCDR3 having an amino acid sequence as shown in SEQ ID NO: 12, 20 or 42; the light chain variable region comprises LCDR1 having an amino acid sequence as shown in SEQ ID NO: 13 or 21; the light chain variable region comprises LCDR2 having an amino acid sequence as shown in SEQ ID NO: 14 or 22; and the light chain variable region comprises LCDR3 having an amino acid sequence as shown in SEQ ID NO: 15 or 23. The antibody of the present invention can inhibit TL1A-induced IFN-γ and TNF-α secretion by PBMCs, and can inhibit TL1A-induced apoptosis, thus prompting that the antibody or a composition thereof can be used for treating diseases such as inflammatory bowel disease, and gastrointestinal diseases associated with cystic fibrosis.

Description

An anti-TL1A antibody and its preparation method and use Technical Field

[0001] The present invention relates to the field of antibody drugs, and in particular to an anti-TL1A antibody and a preparation method and use thereof. Background Art

[0002] Inflammatory bowel disease (IBD) is a specific chronic inflammatory bowel disease that includes Crohn's disease (CD) and ulcerative colitis (UC). The etiology and pathogenesis of IBD remain unclear, but it is generally believed to be the result of the interaction of multiple factors, including dysregulation of intestinal mucosal immunity, persistent intestinal infection, intestinal mucosal barrier defects, and genetic and environmental factors. Both innate and adaptive immune responses in the intestinal mucosa contribute to the pathogenesis of IBD, and abnormal immune responses in the intestinal mucosa are a major cause of mucosal tissue damage.

[0003] TL1A (also known as tumor necrosis factor superfamily member 15 [TNFSF15]) is a member of the TNF ligand superfamily and, like other members, is a homotrimer. TL1A is a type II transmembrane protein that exists in both membrane-bound and soluble forms. Its receptors include DR3 and soluble decoy receptor 3 (DcR3). DR3, also known as APO3, LARD, or WSL1, is a member of the tumor necrosis factor receptor superfamily (TNFRSF). TL1A is the ligand for DR3. Binding to TL1A induces activation of nuclear factor-κB (NF-κB), promoting the transcription of inflammatory factors and activating apoptotic proteases, thereby exerting both proinflammatory and proapoptotic effects. DcR3 competitively binds to TL1A, thereby attenuating T cell costimulatory signals, exerting anti-apoptotic effects and inhibiting the secretion of proinflammatory factors.

[0004] Recent studies have found that TL1A can promote intestinal mucosal inflammation and fibrosis by influencing the activation and proliferation of T cells in the intestinal epithelial lamina propria, promoting the polarization and effector function of Th1 cells, and upregulating IFN-γ expression. Furthermore, TL1A can act on Th17 cells to increase IL-17 secretion, leading to immune dysregulation in the intestinal mucosa and inducing intestinal inflammatory responses. Chronic inflammation is the primary factor in the development of intestinal fibrosis. Inflammation can further activate intestinal fibroblasts, producing large amounts of ECM, leading to the formation of intestinal fibrosis. Therefore, TL1A may promote the development of chronic experimental colitis-associated intestinal fibrosis by regulating IL-17 and IFN-γ. Anti-TL1A antibodies can effectively inhibit TL1A-induced secretion of IFN-γ and TNF-α from human peripheral blood mononuclear cells (PBMCs) and inhibit TL1A-induced apoptosis, thus potentially being used to treat inflammatory bowel disease and related conditions. Summary of the Invention

[0005] In view of the above-mentioned shortcomings of the prior art, the object of the present invention is to provide an anti-TL1A antibody and a preparation method and use thereof, so as to solve the problems in the prior art.

[0006] To achieve the above objectives and other related objectives, the present invention provides an anti-TL1A antibody or an antigen-binding fragment thereof, wherein the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, and wherein the anti-TL1A antibody or the antigen-binding fragment thereof has one or more of the following technical features:

[0007] <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 10 or 18;

[0008] <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO: 11, 19, 39, 40 or 41;

[0009] <3> The heavy chain variable region includes HCDR3 with the amino acid sequence shown in SEQ ID NO: 12, 20 or 42;

[0010] <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 13 or 21;

[0011] <5> The light chain variable region includes LCDR2 with the amino acid sequence shown in SEQ ID NO: 14 or 22;

[0012] <6> The light chain variable region includes LCDR3 having an amino acid sequence as shown in SEQ ID NO: 15 or 23.

[0013] The present invention also provides a recombinant protein comprising: (i) the anti-TL1A antibody or the antigen-binding fragment thereof; and (ii) an optional tag sequence for facilitating expression and / or purification.

[0014] The present invention also provides an isolated polynucleotide encoding the heavy chain variable region and / or light chain variable region or full-length amino acids of the anti-TL1A antibody or antigen-binding fragment thereof, or encoding the recombinant protein.

[0015] The present invention also provides a nucleic acid construct comprising the isolated polynucleotide.

[0016] The present invention also provides a cell, wherein the cell contains the construct or the exogenous polynucleotide is integrated into the genome.

[0017] The present invention also provides a method for preparing the anti-TL1A antibody or antigen-binding fragment thereof, comprising the steps of: culturing the cells under conditions suitable for expressing the TL1A antibody or antigen-binding fragment thereof, thereby expressing the TL1A antibody or antigen-binding fragment thereof; further, the method further comprises the step of purifying and / or isolating the TL1A antibody or antigen-binding fragment thereof.

[0018] The present invention also provides use of the anti-TL1A antibody or antigen-binding fragment thereof or the recombinant protein in preparing a drug for treating a disease or preparing a drug for diagnosing a disease.

[0019] In the present invention, the disease is selected from any one or more of inflammatory bowel disease, gastrointestinal diseases associated with cystic fibrosis, colitis, irritable bowel syndrome, eosinophilic esophagitis, atopic dermatitis, eczema, scleroderma, arthritis, or rheumatoid arthritis, wherein the inflammatory bowel disease is Crohn's disease or ulcerative colitis.

[0020] The present invention also provides a pharmaceutical composition comprising the anti-TL1A antibody or antigen-binding fragment thereof or the recombinant protein.

[0021] The present invention also provides a method for treating a disease, comprising administering the anti-TL1A antibody or antigen-binding fragment thereof, the recombinant protein or the pharmaceutical composition of the present invention to a subject in need thereof.

[0022] As described above, the anti-TL1A antibodies or antigen-binding fragments thereof of the present invention have the following beneficial effects: they can effectively inhibit the secretion of IFN-γ and TNF-α by human peripheral blood mononuclear cells (PBMCs) induced by TL1A, and can inhibit cell apoptosis induced by TL1A, suggesting that the antibodies or compositions thereof can be used to treat inflammatory bowel disease, gastrointestinal diseases associated with cystic fibrosis, Crohn's disease, colitis, ulcerative colitis, irritable bowel syndrome, eosinophilic esophagitis, atopic dermatitis, eczema, scleroderma, arthritis or rheumatoid arthritis.

[0023] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] FIG1A to FIG1C show the binding ability of the tested mouse antibodies to the target antigen human TL1A-His in Example 1. FIG1A is a graph showing the binding ability of the tested mouse antibodies to the target antigen human TL1A-His in Example 1.

[0025] FIG2A to FIG2C show the results of the blocking activity assay of the tested mouse antibodies in Example 1 against the binding of the stably transfected strain 293FT-DR3 to the TL1A protein.

[0026] FIG3A shows the affinity ELISA test results of the candidate antibodies obtained after the optimization of the physicochemical properties of humanized antibodies 72851, 31122 and humanized antibody 31122 in Example 4, binding to the antigen TL1A-his.

[0027] FIG3B shows the capillary isoelectric focusing test results of the candidate antibodies obtained after the physicochemical properties of humanized antibodies 72851, 31122 and humanized antibody 31122 were optimized in Example 4. ...

[0028] FIG4 shows the results of the experiment in Example 5 in which anti-TL1A antibodies blocked the binding of TL1A to DR3.

[0029] FIG5 shows the experimental results of Example 6 in which the anti-TL1A antibody inhibits the NF-κB signaling pathway of the TL1A TF-1 cell line.

[0030] FIG6 shows the results of the anti-TL1A antibody inhibiting TL1A-induced apoptosis of TF-1 cells in Example 7.

[0031] FIG. 7 shows the results of the anti-TL1A antibody in Example 8 inhibiting the secretion of IFNγ by cells stimulated by TL1A.

[0032] FIG8 shows the results of Example 9 in which anti-TL1A antibodies inhibited TL1A-stimulated cell secretion of TNFα.

[0033] FIG9A shows the affinity ELISA test results of the humanized antibody 72581 in Example 10 binding to TL1A proteins of human, cynomolgus monkey, rat and mouse.

[0034] FIG9B to FIG9E show the efficacy results of the anti-TL1A antibody in Example 10 in the DSS-induced rat chronic inflammatory bowel disease model.

[0035] FIG10A and FIG10B show the efficacy of the anti-TL1A antibody in Example 11 in the TNBS-induced acute inflammatory bowel disease model in rats. DETAILED DESCRIPTION

[0036] The present invention provides an anti-TL1A antibody or an antigen-binding fragment thereof, wherein the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, and wherein the anti-TL1A antibody or the antigen-binding fragment thereof has one or more of the following technical features:

[0037] <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 10 or 18;

[0038] <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO: 11, 19, 39, 40 or 41;

[0039] <3> The heavy chain variable region includes HCDR3 with the amino acid sequence shown in SEQ ID NO: 12, 20 or 42;

[0040] <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 13 or 21;

[0041] <5> The light chain variable region includes LCDR2 with the amino acid sequence shown in SEQ ID NO: 14 or 22;

[0042] <6> The light chain variable region includes LCDR3 having an amino acid sequence as shown in SEQ ID NO: 15 or 23.

[0043] In certain embodiments of the present invention, the anti-TL1A antibody or antigen-binding fragment thereof comprises:

[0044] <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 10;

[0045] <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO: 11;

[0046] <3> The heavy chain variable region includes HCDR3 with the amino acid sequence shown in SEQ ID NO: 12;

[0047] <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 13;

[0048] <5> The light chain variable region includes LCDR2 with the amino acid sequence shown in SEQ ID NO: 14;

[0049] <6> The light chain variable region includes LCDR3 having the amino acid sequence shown in SEQ ID NO:15.

[0050] In certain embodiments of the present invention, the anti-TL1A antibody or antigen-binding fragment thereof comprises:

[0051] <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 18;

[0052] <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO: 19;

[0053] <3> The heavy chain variable region includes HCDR3 with the amino acid sequence shown in SEQ ID NO: 20;

[0054] <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 21;

[0055] <5> The light chain variable region includes LCDR2 with the amino acid sequence set forth in SEQ ID NO: 22;

[0056] <6> The light chain variable region includes LCDR3 having the amino acid sequence shown in SEQ ID NO:23.

[0057] In certain embodiments of the present invention, the anti-TL1A antibody or antigen-binding fragment thereof comprises:

[0058] <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 18;

[0059] <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO: 39;

[0060] <3> The heavy chain variable region includes HCDR3 with the amino acid sequence shown in SEQ ID NO: 20;

[0061] <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 21;

[0062] <5> The light chain variable region includes LCDR2 with the amino acid sequence set forth in SEQ ID NO: 22;

[0063] <6> The light chain variable region includes LCDR3 having the amino acid sequence shown in SEQ ID NO:23.

[0064] In certain embodiments of the present invention, the anti-TL1A antibody or antigen-binding fragment thereof comprises:

[0065] <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 18;

[0066] <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO:40;

[0067] <3> The heavy chain variable region includes HCDR3 with the amino acid sequence shown in SEQ ID NO: 20;

[0068] <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 21;

[0069] <5> The light chain variable region includes LCDR2 with the amino acid sequence set forth in SEQ ID NO: 22;

[0070] <6> The light chain variable region includes LCDR3 having the amino acid sequence shown in SEQ ID NO:23.

[0071] In certain embodiments of the present invention, the anti-TL1A antibody or antigen-binding fragment thereof comprises:

[0072] <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 18;

[0073] <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO:41;

[0074] <3> The heavy chain variable region includes HCDR3 with the amino acid sequence shown in SEQ ID NO: 20;

[0075] <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 21;

[0076] <5> The light chain variable region includes LCDR2 with the amino acid sequence set forth in SEQ ID NO: 22;

[0077] <6> The light chain variable region includes LCDR3 having the amino acid sequence shown in SEQ ID NO:23.

[0078] In certain embodiments of the present invention, the anti-TL1A antibody or antigen-binding fragment thereof comprises:

[0079] <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 18;

[0080] <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO: 39;

[0081] <3> The heavy chain variable region includes HCDR3 with the amino acid sequence set forth in SEQ ID NO:42;

[0082] <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 21;

[0083] <5> The light chain variable region includes LCDR2 with the amino acid sequence set forth in SEQ ID NO: 22;

[0084] <6> The light chain variable region includes LCDR3 having the amino acid sequence shown in SEQ ID NO:23.

[0085] In certain embodiments of the present invention, the anti-TL1A antibody or antigen-binding fragment thereof comprises:

[0086] <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 18;

[0087] <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO:40;

[0088] <3> The heavy chain variable region includes HCDR3 with the amino acid sequence set forth in SEQ ID NO:42;

[0089] <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 21;

[0090] <5> The light chain variable region includes LCDR2 with the amino acid sequence set forth in SEQ ID NO: 22;

[0091] <6> The light chain variable region includes LCDR3 having the amino acid sequence shown in SEQ ID NO:23.

[0092] In certain embodiments of the present invention, the anti-TL1A antibody or antigen-binding fragment thereof comprises:

[0093] <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 18;

[0094] <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO:41;

[0095] <3> The heavy chain variable region includes HCDR3 with the amino acid sequence set forth in SEQ ID NO:42;

[0096] <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 21;

[0097] <5> The light chain variable region includes LCDR2 with the amino acid sequence set forth in SEQ ID NO: 22;

[0098] <6> The light chain variable region includes LCDR3 having the amino acid sequence shown in SEQ ID NO:23.

[0099] CDRs (complementarity determining regions) typically refer to regions within an antibody that are spatially complementary to antigenic determinants. The variability within an antibody is typically not evenly distributed throughout its variable region. Both the heavy and light chain variable regions of monoclonal antibodies typically have three hypervariable regions (HVRs), which are spatially complementary to antigenic determinants. These regions are therefore also known as complementarity determining regions (CDRs). The heavy chain variable region typically includes three CDRs, namely HCDR1, HCDR2, and HCDR3, while the light chain variable region typically includes three CDRs, namely LCDR1, LCDR2, and LCDR3.

[0100] In certain embodiments of the present invention, the heavy chain variable region and the light chain variable region may further include a framework region, and the framework region may be located between the complementary determining regions or at both ends of the complementary determining regions. In certain specific embodiments of the present invention, the sequence of the framework region is a human monoclonal antibody variable region, or a framework region sequence of a murine monoclonal antibody variable region that is substituted, deleted, or added with one or more (specifically, 1-50, 1-30, 1-20, 1-10, 1-5, or 1-3) amino acids. The framework region sequence and the framework region sequence of the human monoclonal antibody variable region sequence may have 80%, 85%, 90%, 93%, 95%, 97%, or 99% or more homology.

[0101] In certain embodiments of the present invention, the amino acid sequence of the heavy chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in any one of SEQ ID NO: 3, SEQ ID NO: 16, SEQ ID NO: 7, SEQ ID NO: 24, and SEQ ID NOs: 26-31.

[0102] In certain embodiments of the present invention, the amino acid sequence of the light chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO:5, SEQ ID NO:17, SEQ ID NO:9, or SEQ ID NO:25.

[0103] In certain embodiments of the present invention, the amino acid sequence of the heavy chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 3, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 5.

[0104] In certain embodiments of the present invention, the amino acid sequence of the heavy chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 16, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 17.

[0105] In certain embodiments of the present invention, the amino acid sequence of the heavy chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO:7, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO:9.

[0106] In certain embodiments of the present invention, the amino acid sequence of the heavy chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 24, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 25.

[0107] In certain embodiments of the present invention, the amino acid sequence of the heavy chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 26, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 25.

[0108] In certain embodiments of the present invention, the amino acid sequence of the heavy chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 27, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 25.

[0109] In certain embodiments of the present invention, the amino acid sequence of the heavy chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 28, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 25.

[0110] In certain embodiments of the present invention, the amino acid sequence of the heavy chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 29, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 25.

[0111] In certain embodiments of the present invention, the amino acid sequence of the heavy chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 30, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 25.

[0112] In certain embodiments of the present invention, the amino acid sequence of the heavy chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO:31, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO:25.

[0113] In certain embodiments of the present invention, the anti-TL1A antibody or antigen-binding fragment thereof comprises a monomer, a bivalent antibody, and / or a multivalent antibody.

[0114] In certain embodiments of the present invention, the antigen-binding fragment is selected from the group consisting of scFv, Fab, Fab', F(ab')2, Fv fragment, heavy chain antibody, and disulfide-linked Fv (dsFv).

[0115] In certain embodiments of the present invention, the heavy chain constant region of the anti-TL1A antibody or antigen-binding fragment thereof is selected from the heavy chain constant region of human IgG1, IgG2, IgG3 or IgG4; preferably, it is the heavy chain constant region of human IgG1.

[0116] In certain embodiments of the present invention, the light chain constant region of the anti-TL1A antibody or antigen-binding fragment thereof is selected from the constant region of a human antibody κ chain or λ chain; preferably, it is a κ chain constant region.

[0117] The amino acid sequence of the heavy chain variable region or light chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof further includes a derivative sequence that optionally includes at least one amino acid sequence added, deleted, modified, and / or substituted with the aforementioned sequence, and that retains binding affinity for TL1A. In another preferred embodiment, a derivative sequence includes 1-5, for example, 1, 2, or 3, amino acids added, deleted, modified, and / or substituted with the aforementioned amino acid sequence, such that the derivative antibody composed of the VH and VL sequences containing the derived CDR sequences retains binding affinity for TL1A.

[0118] The anti-TL1A antibody or antigen-binding fragment thereof is selected from a whole antibody, a single-chain antibody or an antibody fragment.

[0119] A complete antibody includes a variable region (V) and a constant region (C). The constant region includes a light chain constant region (LC) and a heavy chain constant region (HC). The constant region can be a native sequence constant region or a variant of its amino acid sequence. The native sequence constant region is, for example, a constant region of a mammalian, such as a human, native sequence. In certain embodiments of the present invention, the heavy chain constant region is an IgG1 constant region, and / or the light chain constant region is a kappa chain constant region (i.e., a κ chain constant region). In certain embodiments of the present invention, the amino acid sequence of the heavy chain constant region is as shown in SEQ ID NO: 33, and / or the amino acid sequence of the light chain constant region is as shown in SEQ ID NO: 34.

[0120] "Antibody fragments" comprise a portion of a complete antibody, preferably comprising its antigen binding region or variable region. For example, antibody fragments include Fab, Fab', F(ab'), F(ab')2 and Fv fragments. Fv fragments are antibody fragments that contain complete antigen recognition and binding sites. This region consists of a heavy chain variable region and a light chain variable region that are tightly linked to each other, and this connection can be covalent (such as in scFv). In such a conformation, the three CDRs of each variable region interact to determine the antigen binding site on the surface of the VV dimer. The "Fab" fragment includes the variable region and constant region of the light chain, and the variable region and the first constant region (CH1) of the heavy chain. A F(ab') antibody fragment includes a pair of Fab fragments, which are usually covalently linked near the carboxyl terminus via a hinge cysteine ​​between them.

[0121] The present invention also provides a recombinant protein comprising: (i) the anti-TL1A antibody or the antigen-binding fragment thereof; and (ii) an optional tag sequence for facilitating expression and / or purification.

[0122] In certain embodiments of the present invention, the tag sequence includes an Fc tag, an HA tag, a GGGS sequence, a FLAG tag, a Myc tag, a 6His tag, or a combination thereof.

[0123] In certain embodiments of the present invention, the recombinant protein comprises a fusion protein.

[0124] In certain embodiments of the present invention, the recombinant protein is a monomer, a dimer, or a multimer.

[0125] The present invention also provides an isolated polynucleotide encoding the heavy chain variable region and / or light chain variable region or full-length amino acids of the anti-TL1A antibody or antigen-binding fragment thereof, or encoding the recombinant protein.

[0126] In certain embodiments of the present invention, the polynucleotide is selected from RNA (such as mRNA) and DNA (such as cDNA).

[0127] In certain embodiments of the present invention, the polynucleotide sequence encoding the heavy chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 35, SEQ ID NO: 37, or SEQ ID NOs: 43-48.

[0128] In certain embodiments of the present invention, the polynucleotide sequence encoding the light chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:36, or SEQ ID NO:38.

[0129] The present invention also provides a nucleic acid construct comprising the isolated polynucleotide.

[0130] In certain embodiments of the present invention, the nucleic acid construct is selected from the group consisting of DNA, RNA, viral vectors, plasmids, transposons, other gene transfer systems, or combinations thereof.

[0131] In certain embodiments of the present invention, the nucleic acid construct comprises a viral vector, such as a lentivirus, adenovirus, AAV virus, retrovirus, or a combination thereof.

[0132] In certain embodiments of the present invention, the nucleic acid construct is a plasmid, a retrovirus, or a lentiviral vector.

[0133] In certain embodiments of the present invention, the nucleic acid construct is selected from the following group: pTomo lentiviral vector, plenti, pLVTH, pLJM1, pHCMV, pLBS.CAG, pHR, pLV, etc.

[0134] In certain embodiments of the present invention, the nucleic acid construct further comprises an element selected from the group consisting of a promoter, a transcription enhancing element WPRE, a long terminal repeat sequence LTR, and the like.

[0135] The term "nucleic acid construct" refers to an artificially constructed nucleic acid segment that can be introduced into target cells or tissues. The nucleic acid construct can be various expression vectors, each comprising a vector backbone, i.e., an empty vector, and an expression cassette. The term "expression cassette" refers to a sequence that has the potential to encode a protein.

[0136] The type of expression vector is not specifically limited. An expression vector refers to a nucleic acid molecule that allows the insertion of exogenous nucleotides without disrupting the vector's ability to replicate and / or integrate in a host cell. An expression vector may include a nucleic acid sequence that allows it to replicate in a host cell, such as an origin of replication. An expression vector may also include one or more selectable marker genes and other genetic factors. An expression vector is a vector that contains the necessary regulatory sequences to enable transcription and translation of an inserted gene or genes. The expression vector is selected from a eukaryotic expression vector or a prokaryotic expression vector.

[0137] The prokaryotic expression vector is selected from an Escherichia coli expression vector, a Bacillus subtilis expression vector, or a Streptomyces expression vector. In a preferred embodiment, the prokaryotic expression vector is selected from an Escherichia coli expression vector. Compared with other expression systems, the Escherichia coli expression system has a clear genetic background, a short culture cycle, a high expression level of the target gene, and strong anti-pollution ability. The Escherichia coli expression vector is, for example, a pET expression vector, specifically pET28a or pET32a, which can be stably expressed in Escherichia coli. The expression vector can also be a pCW expression vector or a pUC expression vector.

[0138] The eukaryotic expression vector is selected from a yeast expression vector, an insect expression vector, or a mammalian expression vector. The mammalian expression vector is selected from a retroviral expression vector, a lentiviral expression vector, an adenoviral expression vector, or an adeno-associated viral expression vector. In a preferred embodiment, the eukaryotic expression vector is selected from a retroviral expression vector, which can be stably expressed in a cell line, such as pMSCV.

[0139] The host cell is selected from a eukaryotic host cell or a prokaryotic host cell. The eukaryotic host cell is selected from fungi such as yeast, insects, birds, plants, C.elegans or nematodes or mammalian host cells. A non-limiting example of an insect cell is a Spodoptera frugiperda (Sf) cell. The example of a yeast host cell is Saccharomyces cerevisiae (S.cerevisiae), Kluyveromyces lactis (Kluyveromyces lactis, K.lactis) or Yarrowia lipolytica (Yarrowia lipolytica). The example of a mammalian cell is a COS cell, a baby hamster kidney cell, a mouse L cell, a LNCaP cell, a Chinese hamster ovary (CHO) cell, a human embryonic kidney (HEK) cell, an African green monkey cell, a CV1 cell, a Vero or a Hep-2 cell. Examples of prokaryotic host cells include bacterial cells, such as E. coli, Streptomyces, B. subtilis, Salmonella typhi, or mycobacteria.

[0140] Those skilled in the art can transfect the expression vector into host cells according to methods well known in the art to obtain cells containing the gene encoding the anti-TL1A antibody or antigen-binding fragment thereof. For example, the expression vector can be introduced into eukaryotic cells by calcium phosphate coprecipitation, electroporation, microinjection, lipofection, or transfection using a polyamine transfection reagent.

[0141] The present invention also provides a cell, wherein the cell contains the construct or the exogenous polynucleotide is integrated into the genome.

[0142] The cell of the present invention is obtained by transforming the nucleic acid construct into a host cell.

[0143] In certain embodiments of the invention, the cell comprises a prokaryotic cell or a eukaryotic cell.

[0144] In certain embodiments of the present invention, the cell is selected from the group consisting of Escherichia coli, yeast cells, and mammalian cells.

[0145] The present invention also provides a method for preparing the anti-TL1A antibody or antigen-binding fragment thereof, comprising the steps of: culturing the cells under conditions suitable for expressing the TL1A antibody or antigen-binding fragment thereof, thereby expressing the TL1A antibody or antigen-binding fragment thereof; further, the method further comprises the step of purifying and / or isolating the TL1A antibody or antigen-binding fragment thereof.

[0146] The present invention also provides use of the anti-TL1A antibody or antigen-binding fragment thereof or the recombinant protein in preparing a drug for treating a disease or preparing a drug for diagnosing a disease.

[0147] In the present invention, the disease is selected from any one or more of inflammatory bowel disease, gastrointestinal diseases associated with cystic fibrosis, colitis, irritable bowel syndrome, eosinophilic esophagitis, atopic dermatitis, eczema, scleroderma, arthritis, or rheumatoid arthritis, wherein the inflammatory bowel disease is Crohn's disease or ulcerative colitis.

[0148] The present invention also provides a pharmaceutical composition comprising the anti-TL1A antibody or antigen-binding fragment thereof or the recombinant protein of the present invention.

[0149] In certain embodiments of the present invention, a pharmaceutical composition comprises the anti-TL1A antibody or antigen-binding fragment thereof or the recombinant protein of the present invention, and a pharmaceutically acceptable carrier or excipient.

[0150] "Pharmaceutically acceptable" means that the drugs do not produce adverse, allergic or other untoward reactions when properly administered to animals or humans.

[0151] "Pharmaceutically acceptable carriers or excipients" should be compatible with the active ingredient, that is, they can be mixed with it without significantly reducing the efficacy of the drug under normal circumstances. Specific examples of substances that can serve as pharmaceutically acceptable carriers or excipients include sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium methylcellulose, ethylcellulose, and methylcellulose; tragacanth powder; malt; gelatin; talc; solid lubricants such as stearic acid and magnesium stearate; calcium sulfate; vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and cocoa butter; polyols such as propylene glycol, glycerol, sorbitol, mannitol, and polyethylene glycol; alginic acid; emulsifiers such as Tween; wetting agents such as sodium lauryl sulfate; colorants; flavorings; tableting agents, stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic saline solution; and phosphate buffer, etc. These materials are used as needed to aid in the stability of the formulation or to help increase the active or its bioavailability or to produce an acceptable taste or flavor in the case of oral administration.

[0152] The form of the pharmaceutical composition is not particularly limited and can be in the form of solid, liquid, gel, semi-fluid, aerosol or other substances.

[0153] The pharmaceutical composition is mainly targeted at mammals. The mammals are preferably rodents, artiodactyls, perissodactyls, lagomorphs, primates, etc. The primates are preferably monkeys, apes or humans.

[0154] The prepared pharmaceutical composition can be administered by conventional routes, including (but not limited to): intravenous injection, intravenous drip, subcutaneous injection, local injection, intramuscular injection, intratumor injection, intraperitoneal injection (such as intraperitoneal injection), intracranial injection or intracavity injection.

[0155] The present invention also provides a method for treating a disease, comprising administering the anti-TL1A antibody or antigen-binding fragment thereof, the recombinant protein or the pharmaceutical composition of the present invention to a subject in need thereof.

[0156] In certain embodiments of the present invention, the subject comprises a mammal, such as a human.

[0157] In certain embodiments of the present invention, the disease is selected from any one or more of inflammatory bowel disease, gastrointestinal disease associated with cystic fibrosis, colitis, irritable bowel syndrome, eosinophilic esophagitis, atopic dermatitis, eczema, scleroderma, arthritis, or rheumatoid arthritis, wherein the inflammatory bowel disease is Crohn's disease or ulcerative colitis.

[0158] In certain embodiments of the present invention, the methods may be used in combination with other therapeutic methods.

[0159] In certain embodiments of the present invention, the other treatment methods include chemotherapy, radiotherapy, targeted therapy and the like.

[0160] When treating a disease, a safe and effective amount of an antibody or antigen-binding fragment thereof is administered to a subject. This safe and effective amount is generally at least about 10 μg / kg body weight, and in most cases does not exceed about 50 mg / kg body weight. Preferably, the dose is about 10 μg / kg body weight to about 10 mg / kg body weight. Of course, the specific dosage will also take into account factors such as the route of administration and the patient's health status, all of which are within the skill of a skilled physician.

[0161] The following describes the embodiments of the present invention through specific examples. Those skilled in the art will readily understand the other advantages and benefits of the present invention from the disclosure herein. The present invention may also be implemented or applied through various other specific embodiments, and the details in this specification may be modified or altered based on different viewpoints and applications without departing from the spirit of the present invention.

[0162] Before further describing the specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific specific embodiments described below; it should also be understood that the terms used in the examples of the present invention are for describing specific specific embodiments rather than for limiting the scope of protection of the present invention; in the present specification and claims, unless otherwise expressly stated herein, the singular forms "a", "an" and "the" include plural forms.

[0163] When the embodiments provide numerical ranges, it should be understood that, unless otherwise specified in the present invention, both endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention are identical with the meanings generally understood by those skilled in the art. Except for the specific methods, equipment, materials used in the embodiments, according to the grasp of the prior art by those skilled in the art and the record of the present invention, any method, equipment and material of the prior art similar or equivalent to the methods, equipment, materials described in the embodiments of the present invention can also be used to realize the present invention. Unless otherwise specified, percentages and parts are weight percentages and parts by weight.

[0164] Example 1 Antigen Immunization of Animals and Preparation and Screening of Hybridomas

[0165] 1.1 Preparation of antigen protein and positive control antibody

[0166] The human TL1A extracellular domain (TL1A-ECD) sequence, used as an antigen, was obtained from the UniProt database (entry: O95150), with the amino acid sequence shown in SEQ ID NO: 1. A 6×His tag was added to its N-terminus and constructed into the pcDNA 3.4 expression vector. The protein was transfected into HEK-293F cells. After 5 days of expression, the cell culture supernatant was collected and purified to obtain the TL1A-His protein. Similarly, the 6×His tag was replaced with the human IgG1 Fc sequence and transfected into HEK-293F cells. The TL1A-Fc protein was expressed and purified. The positive control antibody pra023 sequence was obtained from patent WO-2021081365-A1.

[0167] 1.2 Immunization of mice

[0168] Female Balb / c mice, 6 to 8 weeks old, were used in the experiment. Following purchase, the mice were housed in a laboratory environment for one week under a 12 / 12 hour light / dark cycle, at 20-25°C and 40%-60% humidity. The human TL1A-His protein expressed in Example 1.1 was routinely immunized three times with 50 μg / mouse / 0.2 ml intraperitoneally (IP) on days 0, 14, and 28. Serum antibody titers were determined by ELISA on days 21 and 35. After three immunizations, mice with high serum antibody titers were challenged with 50 μg / mouse / 0.2 ml of human TL1A-His protein via intraperitoneal (IP) injection. Spleens were harvested 3-4 days later for fusion assays.

[0169] 1.3 Hybridoma cell preparation and screening

[0170] Three to four days after the last immunization of mice, mouse spleen cells were fused with mouse myeloma SP2 / 0 cells using a standard hybridoma technology protocol. The fused cells were evenly suspended in complete culture medium consisting of RPMI1640-GLUMAX supplemented with 1% penicillin-streptomycin, 20% FBS (fetal bovine serum), and 1% HAT. The fused cells were plated at 4 × 10 4 10 cells / 200 μl / well were plated and cultured in 60 96-well culture plates. After 7-12 days, the supernatant was harvested and the hybridoma wells positive for human TL1A binding activity were screened by ELISA.

[0171] The ELISA method for screening hybridoma wells with positive human TL1A binding activity is as follows: TL1A-His was diluted to 1 μg / ml in PBS buffer, 100 μl / well was added to the plate, and incubated overnight at 4°C; the next day, the supernatant was discarded, and the plate was blocked with 5% skim milk powder at 37°C for 1 hour. The plate was washed three times with PBST and set aside for use; the collected hybridoma supernatant was sequentially added to the blocked plate at 100 μl / well and incubated at 37°C for 1 hour; the plate was washed three times with PBST, and HRP-labeled goat anti-mouse IgG secondary antibody was added and incubated at 37°C for 30 minutes; after washing the plate three times with PBST, any remaining droplets were patted dry on absorbent paper as much as possible, 100 μl of TMB was added to each well, and the plate was incubated at room temperature (20±5°C) in the dark for 5 minutes; the substrate reaction was terminated by adding 50 μl of 2M H2SO4 stop solution to each well, and the OD value was read at 450 nm on a microplate reader to analyze the binding ability of the test antibody to the target antigen TL1A. The hybridoma cell lines obtained by amplification and screening in serum-containing complete medium were centrifuged and replaced with serum-free medium Hybridoma-SFM medium to a cell density of 1-2 × 10 6 / ml, cultured at 8% CO2 and 37°C for one week, and the culture supernatant was obtained by centrifugation and purified by Protein G affinity chromatography to obtain anti-human TL1A monoclonal antibody protein. After screening, a total of 60 hybridoma cell lines were obtained. All antibodies obtained from hybridoma cell lines of the present invention can be obtained through genetic engineering methods.

[0172] 1.4 Binding ability of mouse antibodies to human TL1A-His protein

[0173] The binding ability of mouse antibodies to human TL1A-His protein was determined by enzyme-linked immunosorbent assay (ELISA). The specific method is as follows:

[0174] TL1A-His protein was diluted to 1 μg / ml with PBS buffer, 100 μl / well was added to the plate, and incubated at 4°C overnight; then blocked with 5% skim milk powder and incubated at 37°C for 1 hour; after washing the plate 3 times with PBST, the anti-human TL1A mouse antibody prepared in the laboratory was serially diluted 3-fold from 10 μg / ml in 11 gradients with 1% BSA-PBS buffer, 1% BSA-PBS was used as a blank control, and 100 μl / well was added to the pre-coated TL1A-His plate and incubated at 37°C for 1 hour; the plate was washed 3 times with PBST, HRP-labeled goat anti-mouse IgG secondary antibody was added, and the plate was placed at 37°C for 30 minutes. After washing the plate 3 times with PBST, the residual droplets were patted dry on absorbent paper as much as possible, 100 μl of TMB was added to each well, and the plate was placed in the dark at room temperature (20±5°C) for 5 minutes, and 50 μl of 2M The substrate reaction was terminated with H2SO4 stop solution, and the OD value was read at 450nm on a microplate reader to analyze the binding ability of the test antibodies to the antigen human TL1A-His. The obtained data were fitted and analyzed by GraphPad Prism 9 software, and the results are shown in Figures 1A, 1B, and 1C. As shown in Figures 1A, 1B, and 1C, most mouse antibodies have good binding activity with the target antigen TL1A-His, and EC 50 As shown in Tables 1-1 to 1-3.

[0175] Table 1-1

[0176] Table 1-2

[0177] Table 1-3

[0178] 1.5 Determination of the blocking activity of mouse antibodies against the binding of stably transfected 293FT-DR3 to TL1A protein

[0179] In this example, the blocking activity of mouse antibodies against 293FT-DR3 binding to human TL1A protein was determined by flow cytometry (Fluorescence activated cell sorting, FACS).

[0180] In this experiment, 293FT-DR3 (an engineered cell line that highly expresses human DR3 constructed using a lentiviral vector in the laboratory) was used as the target cell. TL1A was expressed by Alexa Fluor TM 647NHS Ester reagent (thermofisher) was connected to 647 fluorescent label, 293FT-DR3 cells were counted, and cells were washed once with 1% BSA-PBS, and 2×10 5 / well, 100 μl / well was plated, the supernatant was removed by centrifugation, and the purified mouse antibody was serially diluted 3-fold from 12 μg / ml in 11 gradients with 1% BSA-PBS. 1% BSA-PBS was used as a blank control. 100 μl of the antibody dilution solution was mixed with TL1A-his-647 fluorescent protein (homemade, final concentration 100 ng / ml, 20 μl / well) and incubated at 37°C for 30 minutes. Then 100 μl of the mixture was resuspended in 293FT-DR3 cell wells and incubated at 4°C for 1 hour. The cells were washed twice with 1% BSA-PBS and resuspended in 200 μL PBS. The blocking activity of the mouse antibody against the binding of the cell to the TL1A-his protein was determined by flow cytometry. The data were fitted and analyzed by GraphPad Prism 9 software. The results are shown in Figures 2A, 2B, and 2C. The results showed that most mouse antibodies could specifically block the binding of DR3 and TL1A-his protein on the surface of 293FT cells, and the IC 50 As shown in Table 2-1 to Table 2-3.

[0181] Table 2-1

[0182] Table 2-2

[0183] Table 2-3

[0184] Example 2 Humanization of murine anti-human TL1A monoclonal antibody

[0185] 2.1 Determination of the variable region sequence of the mouse anti-human TL1A monoclonal antibody

[0186] Hybridoma clones 728H4D9 and 311C9E10 were selected as candidate antibodies. Total RNA was extracted from the corresponding hybridoma monoclonal cell lines using Trizol (purchased from Life Technologies). The mRNA was reverse transcribed into cDNA using a reverse transcription kit (purchased from Takara). PCR was performed using a primer combination reported in the literature ("Antibody Engineering," Volume 1, Edited by Roland Kontermann and Stefan Dübel, the primer combination sequence is from page 323). The resulting PCR products were sequenced and analyzed using the Kabat database to confirm that the obtained sequences were variable region sequences of murine antibodies. The heavy chain variable region gene sequence of 728H4D9 is 357 bp long, encoding 119 amino acid residues, with the nucleotide sequence shown in SEQ ID NO: 2 and the amino acid sequence shown in SEQ ID NO: 3. The light chain variable region gene sequence is 318 bp long, encoding 106 amino acid residues, with the nucleotide sequence shown in SEQ ID NO: 4 and the amino acid sequence shown in SEQ ID NO: 5. The heavy chain variable region gene sequence of 311C9E10 is 357 bp in length, encoding 119 amino acid residues. The nucleotide sequence is shown in SEQ ID NO:6, and the amino acid sequence is shown in SEQ ID NO:7. The light chain variable region gene sequence is 318 bp in length, encoding 106 amino acid residues. The nucleotide sequence is shown in SEQ ID NO:8, and the amino acid sequence is shown in SEQ ID NO:9.

[0187] 2.2 Humanization of mouse anti-human TL1A monoclonal antibody

[0188] The amino acid sequences of the heavy and light chain variable regions of 728H4D9 were analyzed, and three antigenic complementarity-determining regions (CDRs) and four framework regions (FRs) were identified according to the Kabat rules. The amino acid sequences of the heavy chain CDRs are HCDR1: GYTMN (SEQ ID NO: 10), HCDR2: LINPYSGGTNYNQKFKG (SEQ ID NO: 11), and HCDR3: IYQRHDGIAY (SEQ ID NO: 12), and the amino acid sequences of the light chain CDRs are LCDR1: SASSVNYMH (SEQ ID NO: 13), LCDR2: DTSKLAS (SEQ ID NO: 14), and LCDR3: QQWSSSPYT (SEQ ID NO: 15).

[0189] By performing homology comparison with human IgG germline sequences in NCBI IgBlast, IGHV1-2*02 was selected as the heavy chain CDR grafting template, and IGKV6-21*01 was selected as the light chain CDR grafting template. The CDR regions of the 728H4D9 antibody were transplanted onto the selected humanized templates, replacing the CDR regions of the humanized templates. The heavy chain variable region was then recombined with the human IgG1 constant region (amino acid sequence shown in SEQ ID NO: 33), and the light chain variable region was recombined with the human kappa chain constant region (amino acid sequence shown in SEQ ID NO: 34). At the same time, based on the three-dimensional structure of the antibody, buried residues, residues that directly interact with the CDR region, and residues that have a significant impact on the VL and VH conformations of the antibody were backmutated to obtain multiple humanized antibodies. The amino acid sequences of the heavy chain variable region (SEQ ID NO: 16) and light chain variable region (SEQ ID NO: 37) of the humanized antibody 72851 were determined through affinity screening. NO:17), and the nucleotide sequences are shown in SEQ ID NO:35 and SEQ ID NO:36, respectively.

[0190] The amino acid sequences of the heavy and light chain variable regions of 311C9E10 were analyzed, and three complementarity-determining regions (CDRs) and four framework regions (FRs) were identified according to the Kabat rules. The amino acid sequences of the heavy chain CDRs are HCDR1: NYWMN (SEQ ID NO: 18), HCDR2: GIRLKSNNYTTQYAESVKG (SEQ ID NO: 19), and HCDR3: LLLNGMDY (SEQ ID NO: 20). The amino acid sequences of the light chain CDRs are LCDR1: SASSVSYMH (SEQ ID NO: 21), LCDR2: DTSNLAS (SEQ ID NO: 22), and LCDR3: FQESGYPFT (SEQ ID NO: 23).

[0191] By comparing homology with human IgG germline sequences in NCBI IgBlast, IGHV3-73*01 was selected as the heavy chain CDR grafting template, and IGKV6-21*01 was selected as the light chain CDR grafting template. The CDR regions of the 311C9E10 antibody were transplanted onto the selected humanized templates, replacing the CDR regions of the human templates. The heavy chain variable region was then recombined with the human IgG1 constant region (amino acid sequence shown in SEQ ID NO: 33), and the light chain variable region was recombined with the human kappa chain constant region (amino acid sequence shown in SEQ ID NO: 34). At the same time, based on the three-dimensional structure of the antibody, buried residues, residues that directly interact with the CDR region, and residues that have a significant impact on the VL and VH conformations of the antibody were backmutated to obtain multiple humanized antibodies. Through affinity screening, the heavy chain variable region sequence (SEQ ID NO: 24) and light chain variable region sequence (SEQ ID NO: 35) of the humanized antibody 31122 were determined. NO:25), and the nucleotide sequences are shown in SEQ ID NO:37 and SEQ ID NO:38, respectively.

[0192] Example 3 Determination of affinity dissociation constant KD of humanized antibodies

[0193] Using a Biacore 8K molecular interaction analyzer, we determined the kinetic parameters for binding and dissociation between candidate humanized antibodies 72851 and 31122, as well as the positive control antibody PRA023, and the antigen TL1A-his using a capture method. Antibodies were diluted to 2 μg / ml in HBS-EP+, pH 7.4 buffer and captured using a Protein A chip. Antigens were then diluted in HBS-EP+, pH 7.4 buffer and bound to the antibodies over six concentration gradients, with a maximum concentration of 50 nM. Dissociation was then performed in HBS-EP+, pH 7.4 buffer. The results are shown in Table 3. Candidate humanized antibodies 72851 and 31122 exhibited superior affinity for TL1A-his compared to the positive control antibody PRA023.

[0194] Table 3: Affinity dissociation constants Note: KD is the affinity constant; ka is the association rate constant; kd is the dissociation rate constant.

[0195] Example 4 Optimization of the physicochemical properties of humanized antibody 31122

[0196] Due to the high glycosylation modification and charge heterogeneity of the humanized antibody 31122, a mutant library was constructed by introducing single-site saturation mutations and multiple superimposed dominant mutations at potential glycosylation sites and deamination sites in the CDR sequence of the heavy chain variable region (light chain unchanged, heavy chain constant region unchanged). ELISA affinity and capillary isoelectric focusing assays were used to screen candidate antibodies with better affinity and charge heterogeneity than the parental 31122 antibody.

[0197] The modified heavy chain variable region CDRs of each candidate antibody are shown in Table 4.

[0198] Table 4: CDR amino acid sequences

[0199] The amino acid sequences of the heavy chain variable regions of the modified candidate antibodies are shown in SEQ ID NOs: 26 to 31, and the nucleotide sequences are shown in SEQ ID NOs: 43 to 48, respectively.

[0200] The results of the ELISA experiment are shown in Figure 3A, which show that the affinity of candidate antibodies 72851, 31122, 311V25, 311V31, 311V34, 311V25-7, 311V31-7, and 311V34-7 for binding to the antigen TL1A-his is better than that of the positive control PRA023.

[0201] The capillary isoelectric focusing test results of the anti-TL1A candidate antibodies are shown in Figure 3B. The experimental results show that the charge heterogeneity of the modified 311V25-7 and 311V31-7 antibodies is better than that of their parent antibody 31122 and is comparable to that of the candidate antibody 72851.

[0202] Example 5 Anti-TL1A antibody blocking TL1A and DR3 binding experiment

[0203] In this example, the blocking effect of anti-TL1A antibodies on the binding of TL1A to its cell surface receptor DR3 was determined using flow cytometry (FACS). The target cells used were a laboratory-generated 293FT cell line (293FT-DR3 cell line) that overexpresses human DR3. The DR3 sequence was obtained from the UniProt database (entry: Q93038), and the amino acid sequence is shown in SEQ ID NO: 32.

[0204] TL1A-his antigen was conjugated to Alexa Fluor TM647 fluorescent dye (purchased from Invitrogen, catalog number A20006) was prepared. The antigen TL1A-his-647 conjugated with fluorescent dye was diluted to 200 ng / ml with 1% BSA-PBS. The anti-TL1A antibody was diluted to 200 nM with 1% BSA-PBS, and 8 concentration gradients were formed by 4-fold dilution. 200 ng / ml of TL1A-his-647 was added to the antibody at different concentration gradients, incubated at 4°C for 1 h, and added to the 293FT-DR3 cell plate (1×10 5 The cells were plated in 4% paraformaldehyde (4% paraformaldehyde) and incubated at 4°C for 1 hour. The cells were washed twice with PBS to remove unbound antigen and resuspended in 200 μL of PBS. The mean fluorescence intensity was measured by flow cytometry, and the data were analyzed using GraphPad Prism 9 software. The results are shown in Figure 4. The experimental results showed that the candidate antibodies 72851, 31122, 311V25, 311V31, 311V34, 311V25-7, 311V31-7, 311V34-7, and the positive control antibody PRA023 all blocked the binding of TL1A to DR3, and the blocking ability of the candidate antibodies was superior to that of PRA023.

[0205] Example 6 Experiment on the inhibition of TL1A activation of NF-κB by anti-TL1A antibody

[0206] After TL1A binds to its receptor DR3, it activates the intracellular NF-κB signaling pathway and triggers cell apoptosis. In this example, a TF-1 cell line (TF-1-NF-κB-Luc cells) stably expressing luciferase regulated by NF-κB was constructed to determine the ability of anti-TL1A antibodies to inhibit TL1A-induced NF-κB pathway activation.

[0207] TL1A-his protein was diluted to 800 ng / mL (final concentration 200 ng / mL) with RPMI 1640 + 10% FBS medium and added to a flat-bottom 96-well plate, 25 μL per well. Anti-TL1A antibody was diluted to 200 nM with RPMI 1640 + 10% FBS medium, and 2-fold dilutions were made into 8 concentration gradients. 25 μL was added to the TL1A-his wells, with the anti-TL1A antibody starting at a concentration of 50 nM. TF-1-NF-κB-Luc cells were centrifuged and counted. The cells were added to the above 96-well plate at the adjusted density, and 1×10 5 / well, 50 μL per well, incubated in a 37°C incubator for 24 hours. The next day, the cell plate was removed and allowed to equilibrate at room temperature for about 15 minutes. 100 μL of Bio-Lite Luciferase (purchased from Nanjing Novozymes Biotech Co., Ltd., catalog number: DD1201-02) was added to each well. After incubation at room temperature for 10 minutes, the luminescence (RLU) value was read on a microplate reader. GraphPad Prism 9 was used for data analysis, graphing, and calculation of IC 50 The results are shown in Figure 5. The experimental results show that candidate antibodies 72851, 311V25-7, 311V31-7 and the positive control antibody PRA023 can effectively inhibit TL1A from activating NF-κB, and the inhibitory ability of the candidate antibodies is better than that of the positive control antibody PRA023.

[0208] Example 7 Anti-TL1A antibody inhibits TL1A-induced apoptosis of TF-1 cells

[0209] TL1A is a potent and specific inhibitor of endothelial cell growth that induces apoptosis by binding to DR3. This example uses flow cytometry (Fluorescence activated cell sorting, FACS) to determine whether anti-TL1A antibodies inhibit TL1A-induced apoptosis in TF-1 cells.

[0210] TF-1 cells were centrifuged and counted, and the cell density was adjusted and plated into 3599 cell plates, with 1×10 5100 μL / well. Dilute TL1A-his to 800 ng / mL in 1640 medium (containing 10% FBS and 1% PS). Simultaneously, add cycloheximide (CHX, purchased from CST, Cat. No. 2112) to the centrifuge tube to a final concentration of 40 μg / mL. Mix thoroughly and add 50 μL per well to the 3599 cell plate. Dilute the positive control antibody PRA023 and other humanized TL1A antibodies to 800 nM in medium, then three-fold dilutions to a total of nine concentrations. Add 50 μL per well to the 3599 cell plate. This results in a final TL1A-his concentration of 200 ng / mL, a final CHX concentration of 10 μg / mL, and a starting TL1A antibody concentration of 200 nM. Gently tap the plate to mix thoroughly and place in a 37°C incubator. After 6 hours, the cell plate was removed and centrifuged at 500g for 5 minutes. The supernatant was discarded and washed twice with 1% BSA-PBS. The cells were stained using the Annexin V-FITC / PI apoptosis kit (purchased from Yisheng Biotechnology (Shanghai) Co., Ltd., catalog number: 40302ES60). The cells were resuspended in 1x Binding buffer, 100 μL / well, and 5 μL Annexin V-FITC and 10 μL PI Staining Solution were added to each well. After standing at room temperature in the dark for 10-15 minutes, the cells were mixed evenly and detected using a flow cytometer. GraphPad Prism9 was used for data analysis, plotting, and calculating EC 50 The results are shown in Figure 6. The experimental results showed that the candidate antibodies 311V25-7, 311V31-7, 311V34-7 and the positive control antibody PRA023 can inhibit TL1A-induced apoptosis of TF-1 cells, and the inhibitory ability of the candidate antibodies is better than that of the positive control antibody PRA023.

[0211] Example 8 Anti-TL1A antibodies inhibit TL1A-stimulated cells from secreting IFNγ

[0212] TL1A can stimulate IFN-γ secretion by binding to DR3. This example measured the activity of anti-TL1A antibodies in inhibiting TL1A-induced IFN-γ secretion in PBMCs.

[0213] PBMCs were diluted in RPMI 1640 + 10% FBS medium and the cell density was adjusted to 3 × 10 6100 μl of cell suspension was added to each well. TL1A-his was diluted to 400 ng / ml in culture medium, and the antibody was diluted to 200 nM. Three-fold serial dilutions were performed, with a 1:1 volume ratio of TL1A-his to antibody. After incubation at room temperature for 30 minutes, the mixture was added to the PBMC plate, with 50 μl of the mixture added to each well. Recombinant IL12 protein was diluted to 4 ng / ml in culture medium, with 25 μl added to each well. Recombinant IL18 protein was diluted to 40 ng / ml in culture medium, and 25 μl added to each well. The plate was incubated at 37°C, 5% CO2 for 48 hours, and the cell supernatant was collected and assayed for human IFNγ expression. The results are shown in Figure 7. The experimental results show that candidate antibodies 72851, 311V25-7, and 311V34-7, as well as the positive control antibody PRA023, all inhibited IFN-γ secretion from PBMCs, with the candidate antibodies exhibiting superior inhibitory activity to the positive control PRA023.

[0214] Example 9: Anti-TL1A antibodies inhibit TL1A-stimulated cell secretion of TNFα

[0215] TL1A, as a Th1 polarizing factor, has the activity to stimulate Th1 cells to secrete IFN-γ and TNF-α cytokines, and therefore may play an important role in the development and progression of inflammatory bowel disease (IBD). This example measured the activity of anti-TL1A antibodies in inhibiting TL1A-induced TNF-α secretion in PBMCs.

[0216] PBMCs were diluted in RPMI 1640 + 10% FBS medium and the cell density was adjusted to 3 × 10 6 100 μl of cell suspension was added to each well. TL1A-his was diluted to 400 ng / ml in culture medium, and the antibody was diluted to 200 nM and 80 nM in a three-fold serial dilution. The TL1A-his and antibody mixture was mixed at a 1:1 volume ratio and incubated at room temperature for 30 minutes. The mixture was then added to the PBMC plate, with 50 μl of the mixture added to each well. Recombinant IL23 protein was diluted to 400 ng / ml in culture medium, with 25 μl added to each well. Recombinant IL2 protein was diluted to 80 U / ml in culture medium, with 25 μl added to each well. The plate was incubated at 37°C in a 5% CO2 incubator for 72 hours, and the cell supernatant was collected and assayed for human TNF-α expression. The results are shown in Figure 8. The experimental results demonstrate that candidate antibodies 72851, 311V25-7, and 311V34-7, as well as the positive control antibody PRA023, all inhibit TNF-α secretion from PBMCs, with the inhibitory activity of the candidate antibodies being superior to that of the positive control PRA023.

[0217] Example 10 Study on the efficacy of anti-TL1A antibody in a DSS-induced rat chronic inflammatory bowel disease model

[0218] When DSS is dissolved in rat drinking water, the high negative charge of its sulfate groups disrupts the intestinal mucosal barrier, triggering an inflammatory response. The animals exhibit significant weight loss, loose stools, hematochezia, and granulocyte infiltration, with clinical symptoms and pathological features closely resembling those of human ulcerative colitis. The anti-TL1A antibody candidate 72851 exhibits comparable binding activity to TL1A homologous proteins from humans (human-TL1A), cynomolgus monkeys (with the same sequence as rhesus macaques, Cynomolgus (Rhesus)-TL1A), and rats (Rat-TL1A) (ELISA results shown in Figure 9A). Therefore, in vivo efficacy studies were conducted in a DSS-induced rat model of chronic inflammatory bowel disease.

[0219] The experiment was divided into three groups: a blank control group, a model group, and a 72851-treated group. The blank control group was given normal drinking water. The model and 72851-treated groups were given drinking water containing 5% DSS for five days per week and pure water for two days per week. Each cycle consisted of seven days, and the experiment lasted for four cycles. The dose of 72851 was 5 mg / kg, administered every two days. At the end of the experiment, the animals were anesthetized and blood was drawn to collect serum. After euthanasia, the entire intestine from the cecum to the end of the rectum was dissected and its length measured. The spleen was removed and weighed. Serum levels of IL-6 and IL-1β were measured by ELISA.

[0220] The experimental results showed that compared with the model group (5% DSS), the candidate antibody 72851 in the 72851 treatment group (5% DSS+72851) effectively inhibited the weight loss of rats (as shown in Figure 9B) and the shortening of the colon (as shown in Figure 9C), and effectively inhibited the increase of inflammatory factors IL-6 and IL-1β in rats (as shown in Figures 9D and 9E).

[0221] Example 11 Study on the efficacy of anti-TL1A antibody in TNBS-induced acute inflammatory bowel disease model in rats

[0222] TNBS is a small molecule hapten that, when administered rectally, binds to host proteins and triggers an immune response, leading to transmural necrosis and extensive inflammatory cell infiltration in the colon, accompanied by weight loss and intestinal obstruction. It is a commonly used agent to simulate the clinical acute enteritis model of Crohn's disease in animals. We validated the efficacy of the anti-TL1A antibody 72851 in treating acute enteritis in rats using this model and compared it with Roche's RVT-3101.

[0223] The experiment was divided into four groups: a blank control group, a model group, an RVT-3101-treated group, and a 72851-treated group. Animals were fasted for 18 hours before the experiment began. RVT-3101 and 72851 were administered before modeling at a dose of 5 mg / kg. Then, under anesthesia, 400 μl of a 1.5% TNBS solution in ethanol (50% ethanol) was perfused into the colon via a plastic catheter via the anus in the model, RVT-3101, and 72851-treated groups. The rats were kept in an inverted position for 3 minutes to prevent leakage of the perfusion solution. The animals were observed during anesthesia. Once awake, they were returned to their cages and housed with free access to food and water. RVT-3101 and 72851 were administered intraperitoneally every two days; the model group received PBS injections every two days, with a dosing volume of 5 ml / kg. On day 7 of the experiment, the animals were anesthetized, blood was collected, and the animals were euthanized. The entire intestine from the cecum to the terminal rectum was cut and measured in length, and the spleen was removed and weighed.

[0224] The experimental results showed that compared with the model group (1.5% TNBS), the candidate antibody 72851 in the 72851 treatment group (1.5% TNBS + 72581) effectively inhibited the shortening of the rat colon (as shown in Figure 10A) and the increase in spleen weight (as shown in Figure 10B), and the efficacy was better than that of the positive control RVT-3101 treatment group (1.5% TNBS + RVT3101).

[0225] The above examples are intended to illustrate the embodiments disclosed herein and are not to be construed as limiting the present invention. In addition, the various modifications listed herein and variations of the methods in the invention will be apparent to those skilled in the art without departing from the scope and spirit of the present invention. Although the present invention has been specifically described in conjunction with various specific preferred embodiments of the present invention, it should be understood that the present invention should not be limited to these specific embodiments. In fact, various modifications apparent to those skilled in the art as described above to obtain the invention should be included within the scope of the present invention.

Claims

1. An anti-TL1A antibody or an antigen-binding fragment thereof, characterized in that: The anti-TL1A antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, and the anti-TL1A antibody or antigen-binding fragment thereof has one or more of the following technical features; <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 10 or 18; <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO: 11, 19, 39, 40 or 41; <3> The heavy chain variable region includes HCDR3 with the amino acid sequence shown in SEQ ID NO: 12, 20 or 42; <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 13 or 21; <5> The light chain variable region includes LCDR2 with the amino acid sequence shown in SEQ ID NO: 14 or 22; <6> The light chain variable region includes LCDR3 having an amino acid sequence as shown in SEQ ID NO: 15 or 23.

2. The anti-TL1A antibody or antigen-binding fragment thereof according to claim 1, characterized in that The anti-TL1A antibody or antigen-binding fragment thereof comprises: <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 10; <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO: 11; <3> The heavy chain variable region includes HCDR3 with the amino acid sequence shown in SEQ ID NO: 12; <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 13; <5> The light chain variable region includes LCDR2 with the amino acid sequence shown in SEQ ID NO: 14; <6> The light chain variable region includes LCDR3 with the amino acid sequence set forth in SEQ ID NO: 15; And / or, the anti-TL1A antibody comprises: <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 18; <2> The heavy chain variable region includes HCDR2 with an amino acid sequence as shown in any one of SEQ ID NOs: 19, 39, 40, and 41; <3> The heavy chain variable region includes HCDR3 with the amino acid sequence shown in SEQ ID NO: 20 or 42; <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 21; <5> The light chain variable region includes LCDR2 with the amino acid sequence set forth in SEQ ID NO: 22; <6> The light chain variable region includes LCDR3 having the amino acid sequence shown in SEQ ID NO:

23.

3. The anti-TL1A antibody or antigen-binding fragment thereof according to claim 1 or 2, characterized in that: The anti-TL1A antibody or antigen-binding fragment thereof further comprises: <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 18; <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO: 19; <3> The heavy chain variable region includes HCDR3 with the amino acid sequence shown in SEQ ID NO: 20; <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 21; <5> The light chain variable region includes LCDR2 with the amino acid sequence set forth in SEQ ID NO: 22; <6> The light chain variable region includes LCDR3 with the amino acid sequence set forth in SEQ ID NO: 23; And / or, the anti-TL1A antibody or antigen-binding fragment thereof comprises: <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 18; <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO: 39; <3> The heavy chain variable region includes HCDR3 with the amino acid sequence shown in SEQ ID NO: 20; <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 21; <5> The light chain variable region includes LCDR2 with the amino acid sequence set forth in SEQ ID NO: 22; <6> The light chain variable region includes LCDR3 with the amino acid sequence set forth in SEQ ID NO: 23; And / or, the anti-TL1A antibody or antigen-binding fragment thereof comprises: <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 18; <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO:40; <3> The heavy chain variable region includes HCDR3 with the amino acid sequence shown in SEQ ID NO: 20; <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 21; <5> The light chain variable region includes LCDR2 with the amino acid sequence set forth in SEQ ID NO: 22; <6> The light chain variable region includes LCDR3 with the amino acid sequence set forth in SEQ ID NO: 23; And / or, the anti-TL1A antibody or antigen-binding fragment thereof comprises: <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 18; <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO:41; <3> The heavy chain variable region includes HCDR3 with the amino acid sequence shown in SEQ ID NO: 20; <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 21; <5> The light chain variable region includes LCDR2 with the amino acid sequence set forth in SEQ ID NO: 22; <6> The light chain variable region includes LCDR3 with the amino acid sequence set forth in SEQ ID NO: 23; And / or, the anti-TL1A antibody or antigen-binding fragment thereof comprises: <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 18; <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO: 39; <3> The heavy chain variable region includes HCDR3 with the amino acid sequence set forth in SEQ ID NO:42; <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 21; <5> The light chain variable region includes LCDR2 with the amino acid sequence set forth in SEQ ID NO: 22; <6> The light chain variable region includes LCDR3 with the amino acid sequence set forth in SEQ ID NO: 23; And / or, the anti-TL1A antibody or antigen-binding fragment thereof comprises: <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 18; <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO:40; <3> The heavy chain variable region includes HCDR3 with the amino acid sequence set forth in SEQ ID NO:42; <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 21; <5> The light chain variable region includes LCDR2 with the amino acid sequence set forth in SEQ ID NO: 22; <6> The light chain variable region includes LCDR3 with the amino acid sequence set forth in SEQ ID NO: 23; And / or, the anti-TL1A antibody or antigen-binding fragment thereof comprises: <1> The heavy chain variable region includes HCDR1 with the amino acid sequence shown in SEQ ID NO: 18; <2> The heavy chain variable region includes HCDR2 with the amino acid sequence shown in SEQ ID NO:41; <3> The heavy chain variable region includes HCDR3 with the amino acid sequence set forth in SEQ ID NO:42; <4> The light chain variable region includes LCDR1 with the amino acid sequence shown in SEQ ID NO: 21; <5> The light chain variable region includes LCDR2 with the amino acid sequence set forth in SEQ ID NO: 22; <6> The light chain variable region includes LCDR3 having the amino acid sequence shown in SEQ ID NO:

23.

4. The anti-TL1A antibody or antigen-binding fragment thereof according to claim 1, wherein The amino acid sequence of the heavy chain variable region of the anti-TL1A antibody or its antigen-binding fragment is shown in any one of SEQ ID NO:3, SEQ ID NO:16, SEQ ID NO:7, SEQ ID NO:24, and SEQ ID NOs:26-31, and / or the amino acid sequence of the light chain variable region of the anti-TL1A antibody or its antigen-binding fragment is shown in SEQ ID NO:5, SEQ ID NO:17, SEQ ID NO:9, and SEQ ID NO:

25.

5. The anti-TL1A antibody or antigen-binding fragment thereof according to claim 1, wherein Also includes any of the following features: 1) The amino acid sequence of the heavy chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 3, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 5; 2) the anti-TL1A antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence as shown in SEQ ID NO: 16, and a light chain variable region amino acid sequence as shown in SEQ ID NO: 17; 3) the amino acid sequence of the heavy chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 7, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 9; 4) the anti-TL1A antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence as shown in SEQ ID NO: 24, and a light chain variable region amino acid sequence as shown in SEQ ID NO: 25; 5) the anti-TL1A antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence as shown in SEQ ID NO: 26, and a light chain variable region amino acid sequence as shown in SEQ ID NO: 25; 6) the anti-TL1A antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence as shown in SEQ ID NO: 27, and a light chain variable region amino acid sequence as shown in SEQ ID NO: 25; 7) The heavy chain variable region amino acid sequence of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 28, and the light chain variable region amino acid sequence is shown in SEQ ID NO: 25; 8) The amino acid sequence of the heavy chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 29, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 25; 9) The anti-TL1A antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence as shown in SEQ ID NO: 30, and a light chain variable region amino acid sequence as shown in SEQ ID NO: 25; 10) The amino acid sequence of the heavy chain variable region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 31, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO:

25.

6. The anti-TL1A antibody or antigen-binding fragment thereof according to claim 1, wherein The anti-TL1A antibody or antigen-binding fragment thereof is selected from a whole antibody, a single-chain antibody or an antibody fragment; preferably, the heavy chain constant region of the whole antibody is an IgG1 constant region, and / or the light chain constant region is a kappa chain constant region.

7. The anti-TL1A antibody or antigen-binding fragment thereof according to claim 1, wherein The amino acid sequence of the heavy chain constant region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 33; and / or the amino acid sequence of the light chain constant region of the anti-TL1A antibody or antigen-binding fragment thereof is shown in SEQ ID NO:

34.

8. An isolated polynucleotide, characterized in that The polynucleotide encodes the heavy chain variable region and / or light chain variable region or the full-length amino acid sequence of the anti-TL1A antibody or antigen-binding fragment thereof according to any one of claims 1 to 7.

9. The polynucleotide according to claim 8, wherein The polynucleotide sequence encoding the heavy chain variable region of the anti-TL1A antibody or its antigen-binding fragment is shown in SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 35, SEQ ID NO: 37, and SEQ ID NOs: 43 to 48; and / or the polynucleotide sequence encoding the light chain variable region of the anti-TL1A antibody or its antigen-binding fragment is shown in SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 36, and SEQ ID NO:

38.

10. A nucleic acid construct, characterized in that Contains the isolated polynucleotide according to claim 8 or 9.

11. A cell, characterized in that The cell contains the construct according to claim 10 or the exogenous polynucleotide according to claim 8 or 9 is integrated into the genome.

12. The method for preparing the anti-TL1A antibody or antigen-binding fragment thereof according to any one of claims 1 to 7, characterized in that: The method comprises the following steps: culturing the cell according to claim 11 under conditions suitable for expressing the anti-TL1A antibody or the antigen-binding fragment thereof, thereby expressing the anti-TL1A antibody or the antigen-binding fragment thereof.

13. Use of the anti-TL1A antibody or antigen-binding fragment thereof according to any one of claims 1 to 7 in the preparation of a drug for treating a disease or a drug for diagnosing a disease; preferably, the disease is selected from any one or more of inflammatory bowel disease, gastrointestinal diseases associated with cystic fibrosis, colitis, irritable bowel syndrome, eosinophilic esophagitis, atopic dermatitis, eczema, scleroderma, arthritis or rheumatoid arthritis.

14. The use according to claim 13, characterized in that The inflammatory bowel disease is Crohn's disease or ulcerative colitis.

15. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the anti-TL1A antibody or antigen-binding fragment thereof according to any one of claims 1 to 7.

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