Antibodies against tumor necrosis factor-like ligand 1A and uses thereof

Abstract

The invention discloses an antibody combined with a tumor necrosis factor-like ligand 1A (TL1A), a nucleic acid molecule for coding the antibody, a carrier containing the nucleic acid molecule, a host cell containing the nucleic acid molecule or the carrier, a method for preparing and purifying the antibody and application of the antibody.

Description

Technical Field

[0001] This application generally relates to the fields of genetic engineering and antibody drugs; specifically, it relates to antibodies that bind to tumor necrosis factor-like ligand 1A and their uses. Background Art

[0002] Tumor necrosis factor-like ligand 1A (TL1A / VEGI-251 / TNFSF15) is a member of the tumor necrosis factor family, with a full length of 251 amino acids and is a type II transmembrane glycoprotein. As a type II transmembrane protein, TL1A is similar to other members of the TNF family and forms a stable non-covalent homotrimer structure composed of a β-sandwich. TL1A is expressed in immune cells (such as monocytes, macrophages, dendritic cells, T lymphocytes, plasma cells, etc.) and non-immune cells (such as endothelial cells, synovial fibroblasts, etc.). TL1A initially exists in a membrane-bound form and can be cleaved at amino acid position 72 by alternative splicing or tumor necrosis factor α-converting enzyme to release a soluble protein to perform its function. 1 The corresponding functional receptor of TL1A is death receptor 3 (DR3), and the non-functional receptor is decoy receptor 3 (DcR3). 2 TL1A binds to DR3, induces oligomerization of the DR3 receptor, recruits the adaptor protein - tumor necrosis factor receptor-associated death domain protein (TRADD) through the intracellular death domain, and regulates downstream pathways (such as TRAF2, RIP1, PI3K, MAPK, and NF-κB) to exert a pro-inflammatory effect; or promotes apoptosis through the FADD, RIP3, and Caspase-8 / -3 / -7 pathways; meanwhile, the NF-κB pathway can also activate c-IAP proteins, thereby negatively regulating apoptosis. 1,3 。

[0003] The abnormal expression of TL1A is significantly associated with autoimmune diseases, including rheumatoid arthritis, inflammatory bowel disease, psoriasis, primary biliary cirrhosis, systemic lupus erythematosus, and ankylosing spondylitis, etc. 1 The key role of the TL1A-DR3 signaling pathway in autoimmune and inflammatory diseases indicates that inhibiting the TL1A-DR3 interaction may be an effective treatment strategy for improving autoimmune diseases and local inflammation of target organs. Currently developed monoclonal antibodies targeting TL1A have clinical indications including inflammatory bowel disease and systemic sclerosis-related interstitial lung disease, etc. Among them, RVT3101 and PRA023 have both observed significant clinical remission rates and endoscopic improvement rates in clinical trials for the treatment of inflammatory bowel disease.

[0004] Inflammatory bowel disease (IBD) includes Crohn's disease (CD) and ulcerative colitis (UC), which are recurrent and inflammatory diseases involving the digestive tract with intestinal inflammation and epithelial damage as pathological features. The etiology of IBD is not fully understood and may be related to multiple factors such as genetics, environment, intestinal microecology, and intestinal immunity. 4 Current IBD treatment drugs include aminosalicylate preparations, oral glucocorticoids, oral small molecule Janus kinase inhibitors, tumor necrosis factor inhibitors, integrin receptor antagonists, interleukin (IL) 12 / 23 antagonists, etc. 5 However, the above treatment regimens still cannot induce a sufficient response in IBD patients, and there is an unmet clinical need for the treatment of IBD.

[0005] Therefore, the development and application of new anti-human TL1A antibodies are needed in this field and have important biological and medical significance. Summary of the Invention

[0007] In a first aspect, the present application provides an antibody that binds to tumor necrosis factor-like ligand 1A (TL1A), which comprises a heavy chain variable region containing the amino acid sequences of HCDR1, HCDR2, and HCDR3 and a light chain variable region containing the amino acid sequences of LCDR1, LCDR2, and LCDR3, wherein

[0008] the amino acid sequence of the HCDR1 is as shown in SEQ ID NO: 1, the amino acid sequence of the HCDR2 is as shown in SEQ ID NO: 2, the amino acid sequence of the HCDR3 is as shown in SEQ ID NO: 3, the amino acid sequence of the LCDR1 is as shown in SEQ ID NO: 4, the amino acid sequence of the LCDR2 is as shown in SEQ ID NO: 5, and the amino acid sequence of the LCDR3 is as shown in SEQ ID NO: 6; or

[0009] the amino acid sequence of the HCDR1 is as shown in SEQ ID NO: 7, the amino acid sequence of the HCDR2 is as shown in SEQ ID NO: 8, the amino acid sequence of the HCDR3 is as shown in SEQ ID NO: 9, the amino acid sequence of the LCDR1 is as shown in SEQ ID NO: 10, the amino acid sequence of the LCDR2 is as shown in SEQ ID NO: 11, and the amino acid sequence of the LCDR3 is as shown in SEQ ID NO: 12; or

[0010] The amino acid sequence of the HCDR1 is as shown in SEQ ID NO: 7, the amino acid sequence of the HCDR2 is as shown in SEQ ID NO: 13, the amino acid sequence of the HCDR3 is as shown in SEQ ID NO: 9, the amino acid sequence of the LCDR1 is as shown in SEQ ID NO: 14, the amino acid sequence of the LCDR2 is as shown in SEQ ID NO: 15, and the amino acid sequence of the LCDR3 is as shown in SEQ ID NO: 16; or

[0011] The amino acid sequence of the HCDR1 is as shown in SEQ ID NO: 17, the amino acid sequence of the HCDR2 is as shown in SEQ ID NO: 18, the amino acid sequence of the HCDR3 is as shown in SEQ ID NO: 19, the amino acid sequence of the LCDR1 is as shown in SEQ ID NO: 20, the amino acid sequence of the LCDR2 is as shown in SEQ ID NO: 21, and the amino acid sequence of the LCDR3 is as shown in SEQ ID NO: 22;

[0012] Among them, the HCDR and LCDR amino acid sequences are defined according to Kabat.

[0013] In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region of the antibody is as shown in SEQ ID NO: 23, 24, 25 or 26.

[0014] In some embodiments of the first aspect, the amino acid sequence of the light chain variable region of the antibody is as shown in SEQ ID NO: 27, 28, 29 or 30.

[0015] In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region of the antibody is as shown in SEQ ID NO: 23, and the amino acid sequence of the light chain variable region of the antibody is as shown in SEQ ID NO: 27; or

[0016] The amino acid sequence of the heavy chain variable region of the antibody is as shown in SEQ ID NO: 24, and the amino acid sequence of the light chain variable region of the antibody is as shown in SEQ ID NO: 28; or

[0017] The amino acid sequence of the heavy chain variable region of the antibody is as shown in SEQ ID NO: 25, and the amino acid sequence of the light chain variable region of the antibody is as shown in SEQ ID NO: 29; or

[0018] The amino acid sequence of the heavy chain variable region of the antibody is as shown in SEQ ID NO: 26, and the amino acid sequence of the light chain variable region of the antibody is as shown in SEQ ID NO: 30.

[0019] In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region of the antibody has at least 90% identity with SEQ ID NO: 23, 24, 25, or 26, and the amino acid sequence of the light chain variable region of the antibody has at least 90% identity with SEQ ID NO: 27, 28, 29, or 30.

[0020] In some embodiments of the first aspect, the antibody is a whole antibody, Fab fragment, F(ab’) 2 fragment or single-chain Fv fragment (scFv).

[0021] In some embodiments of the first aspect, the antibody is a monoclonal antibody.

[0022] In some embodiments of the first aspect, the antibody further comprises a heavy chain constant region selected from the IgG1 subtype, IgG2 subtype, or IgG4 subtype.

[0023] In some embodiments of the first aspect, the antibody further comprises a light chain constant region selected from the κ subtype or λ subtype.

[0024] In some embodiments of the first aspect, the antibody binds to TL1A of primates and / or rodents; and / or

[0025] the antibody inhibits the binding of TL1A to death receptor 3 (DR3); and / or

[0026] the antibody does not inhibit the binding of TL1A to decoy receptor 3 (DcR3); and / or

[0027] the antibody inhibits the caspase activation of cells stimulated by TL1A; and / or

[0028] the antibody inhibits the NF-κB activation of cells stimulated by TL1A; and / or

[0029] the antibody inhibits the ability of TL1A to stimulate PBMC to secrete interferon γ.

[0030] In a second aspect, the present application provides a nucleic acid molecule encoding the antibody described in the first aspect.

[0031] In a third aspect, the present application provides a pharmaceutical composition comprising the antibody described in the first aspect and a pharmaceutically acceptable excipient, diluent, or carrier.

[0032] In a fourth aspect, the present application provides the use of the antibody described in the first aspect, or the pharmaceutical composition described in the third aspect, in the preparation of a drug for preventing or treating TL1A-related diseases.

[0033] In a fifth aspect, the present application provides a method for preventing or treating TL1A-related diseases, which includes administering to an individual in need the antibody described in the first aspect, or the pharmaceutical composition described in the third aspect. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 The results showing that the anti-TL1A murine monoclonal antibody inhibits the caspase activation in TF-1 cells stimulated by TL1A are presented.

[0035] Figure 2 The results showing that the anti-TL1A murine monoclonal antibody inhibits the NF-κB activation in 293T-hDR3-NF-κB-RE-luci cells stimulated by TL1A are presented.

[0036] Figure 3 The results showing that the anti-TL1A monoclonal antibody H3F1+L28E1 inhibits the caspase activation in TF-1 cells stimulated by TL1A are presented.

[0037] Figure 4 The results showing that the anti-TL1A monoclonal antibody H3F1+L28E1 inhibits the NF-κB activation in 293T-hDR3-NF-κB-RE-luci cells stimulated by TL1A are presented.

[0038] Figure 5 The results showing that the anti-TL1A monoclonal antibody H3F1+L28E1 inhibits the ability of TL1A to stimulate PBMC to secrete interferon γ are presented.

[0039] Figure 6 The ELISA analysis results showing that the anti-human TL1A monoclonal antibody blocks the binding of human TL1A to human DR3 are presented.

[0040] Figure 7 The ELISA analysis results showing that the anti-TL1A monoclonal antibody blocks the binding activity of human DcR3 and human TL1A are presented.

[0041] Figure 8 The results of fecal score in rats with acute ulcerative colitis induced by DNBS are presented.

[0042] Figure 9 The results of macroscopic colon injury score in rats with acute ulcerative colitis induced by DNBS are presented.

[0043] SEQUENCE LISTING

[0044] SEQ ID NO: 1-3 respectively show the amino acid sequences of HCDR1, HCDR2 and HCDR3 of the anti-TL1A single-chain antibody S4D3.

[0045] SEQ ID NO: 4 - 6 show the amino acid sequences of LCDR1, LCDR2, and LCDR3 of the anti - TL1A single - chain antibody S4D3, respectively.

[0046] SEQ ID NO: 7 shows the amino acid sequence of HCDR1 of the anti - TL1A single - chain antibodies S9H9 and S7B11.

[0047] SEQ ID NO: 8 shows the amino acid sequence of HCDR2 of the anti - TL1A single - chain antibody S9H9.

[0048] SEQ ID NO: 9 shows the amino acid sequences of HCDR3 of the anti - TL1A single - chain antibodies S9H9 and S7B11.

[0049] SEQ ID NO: 10 - 12 show the amino acid sequences of LCDR1, LCDR2, and LCDR3 of the anti - TL1A single - chain antibody S9H9, respectively.

[0050] SEQ ID NO: 13 shows the amino acid sequence of HCDR2 of the anti - TL1A single - chain antibody S7B11.

[0051] SEQ ID NO: 14 - 16 show the amino acid sequences of LCDR1, LCDR2, and LCDR3 of the anti - TL1A single - chain antibody S7B11, respectively.

[0052] SEQ ID NO: 17 - 19 show the amino acid sequences of HCDR1, HCDR2, and HCDR3 of the fully human anti - TL1A monoclonal antibody H3F1 + L28E1, respectively.

[0053] SEQ ID NO: 20 - 22 show the amino acid sequences of LCDR1, LCDR2, and LCDR3 of the fully human anti - TL1A monoclonal antibody H3F1 + L28E1, respectively.

[0054] SEQ ID NO: 23 shows the amino acid sequence of the heavy - chain variable region of the anti - TL1A single - chain antibody S4D3.

[0055] SEQ ID NO: 24 shows the amino acid sequence of the heavy - chain variable region of the anti - TL1A single - chain antibody S9H9.

[0056] SEQ ID NO: 25 shows the amino acid sequence of the heavy - chain variable region of the anti - TL1A single - chain antibody S7B11.

[0057] SEQ ID NO: 26 shows the amino acid sequence of the heavy - chain variable region of the fully human anti - TL1A monoclonal antibody H3F1 + L28E1.

[0058] SEQ ID NO:27 shows the amino acid sequence of the light chain variable region of the anti-TL1A single-chain antibody S4D3.

[0059] SEQ ID NO:28 shows the amino acid sequence of the light chain variable region of the anti-TL1A single-chain antibody S9H9.

[0060] SEQ ID NO:29 shows the amino acid sequence of the light chain variable region of the anti-TL1A single-chain antibody S7B11.

[0061] SEQ ID NO:30 shows the amino acid sequence of the light chain variable region of the fully human anti-TL1A monoclonal antibody H3F1+L28E1.

[0062] SEQ ID NO:31 shows the amino acid sequence of recombinant human TL1A (hTL1A).

[0063] SEQ ID NO:32 shows the amino acid sequence of recombinant cynomolgus monkey TL1A (mfTL1A).

[0064] SEQ ID NO:33 shows the amino acid sequence of recombinant mouse TL1A (mTL1A).

[0065] SEQ ID NO:34 shows the amino acid sequence of recombinant rat TL1A (rTL1A).

[0066] SEQ ID NO:35 shows the amino acid sequence of the Foldon domain of the T4 phage fibritin (Fib Foldon).

[0067] SEQ ID NO:36 shows the amino acid sequence of the His tag.

[0068] SEQ ID NO:37 shows the amino acid sequence of the heavy chain constant region of human (homo sapiens) IgG1 subtype (CH-IgG1).

[0069] SEQ ID NO:38 shows the amino acid sequence of the mutant heavy chain constant region of human IgG1 subtype IgG1m3 (CH-IgG1m3).

[0070] SEQ ID NO:39 shows the amino acid sequence of the mutant heavy chain constant region of human IgG1 subtype IgG1m3-YTE (CH-IgG1m3-YTE).

[0071] SEQ ID NO:40 shows the amino acid sequence of the light chain constant region of human (homo sapiens) κ subtype.

[0072] SEQ ID NO:41 shows the amino acid sequence of the constant region of the human (homo sapiens) lambda subtype light chain.

[0073] SEQ ID NO:42 shows the amino acid sequence of the recombinant protein His-fib-hTL1A.

[0074] SEQ ID NO:43 shows the amino acid sequence of the recombinant protein His-fib-mfTL1A.

[0075] SEQ ID NO:44 shows the amino acid sequence of the recombinant protein His-fib-mTL1A.

[0076] SEQ ID NO:45 shows the amino acid sequence of the recombinant protein His-fib-rTL1A.

[0077] SEQ ID NO:46 shows the nucleotide sequence of the primer PmCGR.

[0078] SEQ ID NO:47 shows the nucleotide sequence of the primer PmCKR.

[0079] SEQ ID NO:48 shows the amino acid sequence of the single-chain antibody S4D3.

[0080] SEQ ID NO:49 shows the amino acid sequence of the single-chain antibody S9H9.

[0081] SEQ ID NO:50 shows the amino acid sequence of the single-chain antibody S7B11.

[0082] SEQ ID NO:51 shows the amino acid sequence of the heavy chain variable region of the anti-TL1A monoclonal antibody RVT-3101.

[0083] SEQ ID NO:52 shows the amino acid sequence of the light chain variable region of the anti-TL1A monoclonal antibody RVT-3101.

[0084] SEQ ID NO:53 shows the amino acid sequence of the heavy chain variable region of the anti-TL1A monoclonal antibody PRA023.

[0085] SEQ ID NO:54 shows the amino acid sequence of the light chain variable region of the anti-TL1A monoclonal antibody PRA023.

[0086] SEQ ID NO:55 shows the amino acid sequence of the heavy chain variable region of the negative control antibody DP47.

[0087] SEQ ID NO:56 shows the amino acid sequence of the light chain variable region of the negative control antibody DP47. Detailed Description of the Invention

[0089] The inventors of the present application obtained new antibodies that bind to tumor necrosis factor-like ligand 1A (TL1A) through antibody engineering techniques. In various aspects of the present application, new antibodies against tumor necrosis factor-like ligand 1A (TL1A), nucleic acid molecules encoding the antibodies, vectors containing the nucleic acid molecules, host cells containing the nucleic acid molecules or vectors, methods for preparing and purifying the antibodies, and medical and biological applications of the antibodies are provided. According to the amino acid sequences of the variable regions of the antibodies provided by the present application, full-length antibody molecules can be constructed as drugs for preventing or treating TL1A-related diseases.

[0090] Unless otherwise specified, the implementation of the present application uses conventional molecular biology, microbiology, cell biology, biochemistry, and immunology techniques in the art.

[0091] Unless otherwise specified, the terms used in the present application have the meanings commonly understood by those skilled in the art.

[0092] Definitions

[0093] As used herein, the term "antibody" refers to an immunoglobulin molecule that can specifically bind to a target via at least one antigen recognition site located in the variable region of the immunoglobulin molecule. Targets include, but are not limited to, carbohydrates, polynucleotides, lipids, polypeptides, etc. The "antibody" used herein includes not only intact (i.e., full-length) antibodies, but also antigen-binding fragments thereof (e.g., Fab, Fab’, F(ab’) 2 , Fv), variants thereof, fusion proteins containing antibody portions, humanized antibodies, chimeric antibodies, diabodies, linear antibodies, single-chain antibodies, multispecific antibodies (e.g., bispecific antibodies), and any other modified configurations of immunoglobulin molecules containing the antigen recognition site with the required specificity, including glycosylation variants of antibodies, amino acid sequence variants of antibodies, and covalently modified antibodies.

[0094] Typically, a complete or full-length antibody comprises two heavy chains and two light chains. Each heavy chain contains a heavy chain variable region (VH) and first, second, and third constant regions (CH1, CH2, and CH3). Each light chain contains a light chain variable region (VL) and a constant region (CL). The full-length antibody can be any type of antibody, such as IgD, IgE, IgG, IgA, or IgM (or subclasses thereof), but the antibody need not belong to any particular class. Immunoglobulins can be designated into different classes based on the amino acid sequence of the heavy chain constant domain of the antibody. Typically, there are five main classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these classes can be further differentiated into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to different immunoglobulin classes are designated α, δ, ε, γ, and μ, respectively. The subunit structure and three-dimensional structure of different classes of immunoglobulins are well known.

[0095] As used herein, the term “antigen-binding fragment or antigen-binding portion” refers to a part or region of a complete antibody molecule that is responsible for binding an antigen. The antigen-binding domain can comprise the heavy chain variable region (VH), the light chain variable region (VL), or both. Each of VH and VL typically contains three complementarity-determining regions CDR1, CDR2, and CDR3.

[0096] It is well known to those skilled in the art that the complementarity-determining regions (CDRs, typically CDR1, CDR2, and CDR3) are the regions in the variable region that have the greatest influence on the affinity and specificity of an antibody. There are two common ways to define the CDR amino acid sequences of VH or VL, namely the Chothia definition and the Kabat definition. See, for example, Kabat, “Sequences of Proteins of Immunological Interest”, National Institutes of Health, Bethesda, Md. (1991) 7 ; A1-Lazikani et al., J. Mol. Biol. 273:927-948 (1997) 8 ; and Martin et al., Proc. Natl. Acad. Sci. USA 86:9268-9272 (1989) 9 . For a given variable region amino acid sequence of an antibody, the CDR amino acid sequences in the VH and VL amino acid sequences can be determined according to the Chothia definition or the Kabat definition. In the embodiments of the present application, the Kabat definition is used for the CDR amino acid sequences.

[0097] For the amino acid sequence of the variable region of a given antibody, the CDR amino acid sequences in the variable region amino acid sequence can be analyzed in various ways. For example, the online software Abysis can be used to determine it (http: / / www.abysis.org / ).

[0098] Examples of antigen-binding fragments include, but are not limited to: (1) Fab fragments, which can be monovalent fragments having VL-CL chains and VH-CH1 chains; (2) F(ab’) 2 fragments, which can be divalent fragments having two Fab’ fragments connected by a disulfide bridge in the hinge region (i.e., a dimer of Fab’); (3) Fv fragments having the VL and VH domains of a single arm of an antibody; (4) single-chain Fv (scFv), which can be a single polypeptide chain composed of a VH domain and a VL domain via a peptide linker; and (5) (scFv) 2 which can contain two VH domains and two VL domains connected by a peptide linker, and the two VL domains are combined with the two VH domains via disulfide bridges.

[0099] As used herein, the term “specifically binds” refers to a non-random binding reaction between two molecules, such as the binding of an antibody to an epitope.

[0100] As used herein, the term “monoclonal antibody” refers to an antibody obtained from a substantially homogeneous population of antibodies, i.e., each antibody constituting the population is identical except for possible naturally occurring mutations in a small number of individuals.

[0101] In a first aspect, the present application provides an antibody that binds to tumor necrosis factor-like ligand 1A (TL1A), which comprises a heavy chain variable region containing the amino acid sequences of HCDR1, HCDR2, and HCDR3 and a light chain variable region containing the amino acid sequences of LCDR1, LCDR2, and LCDR3, wherein

[0102] the amino acid sequence of the HCDR1 is as shown in SEQ ID NO: 1, the amino acid sequence of the HCDR2 is as shown in SEQ ID NO: 2, the amino acid sequence of the HCDR3 is as shown in SEQ ID NO: 3, the amino acid sequence of the LCDR1 is as shown in SEQ ID NO: 4, the amino acid sequence of the LCDR2 is as shown in SEQ ID NO: 5, and the amino acid sequence of the LCDR3 is as shown in SEQ ID NO: 6; or

[0103] The amino acid sequence of the HCDR1 is as shown in SEQ ID NO: 7, the amino acid sequence of the HCDR2 is as shown in SEQ ID NO: 8, the amino acid sequence of the HCDR3 is as shown in SEQ ID NO: 9, the amino acid sequence of the LCDR1 is as shown in SEQ ID NO: 10, the amino acid sequence of the LCDR2 is as shown in SEQ ID NO: 11, and the amino acid sequence of the LCDR3 is as shown in SEQ ID NO: 12; or

[0104] The amino acid sequence of the HCDR1 is as shown in SEQ ID NO: 7, the amino acid sequence of the HCDR2 is as shown in SEQ ID NO: 13, the amino acid sequence of the HCDR3 is as shown in SEQ ID NO: 9, the amino acid sequence of the LCDR1 is as shown in SEQ ID NO: 14, the amino acid sequence of the LCDR2 is as shown in SEQ ID NO: 15, and the amino acid sequence of the LCDR3 is as shown in SEQ ID NO: 16; or

[0105] The amino acid sequence of the HCDR1 is as shown in SEQ ID NO: 17, the amino acid sequence of the HCDR2 is as shown in SEQ ID NO: 18, the amino acid sequence of the HCDR3 is as shown in SEQ ID NO: 19, the amino acid sequence of the LCDR1 is as shown in SEQ ID NO: 20, the amino acid sequence of the LCDR2 is as shown in SEQ ID NO: 21, and the amino acid sequence of the LCDR3 is as shown in SEQ ID NO: 22;

[0106] Wherein, the amino acid sequences of HCDR and LCDR are defined according to Kabat.

[0107] In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region of the antibody is as shown in SEQ ID NO: 23, 24, 25 or 26.

[0108] In some embodiments of the first aspect, the amino acid sequence of the light chain variable region of the antibody is as shown in SEQ ID NO: 27, 28, 29 or 30.

[0109] In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region of the antibody is as shown in SEQ ID NO: 23, and the amino acid sequence of the light chain variable region of the antibody is as shown in SEQ ID NO: 27.

[0110] In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region of the antibody is as shown in SEQ ID NO: 24, and the amino acid sequence of the light chain variable region of the antibody is as shown in SEQ ID NO: 28.

[0111] In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region of the antibody is as shown in SEQ ID NO: 25, and the amino acid sequence of the light chain variable region of the antibody is as shown in SEQ ID NO: 29.

[0112] In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region of the antibody is as shown in SEQ ID NO: 26, and the amino acid sequence of the light chain variable region of the antibody is as shown in SEQ ID NO: 30.

[0113] In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region of the antibody has at least 90% identity with SEQ ID NO: 23, 24, 25 or 26, and the amino acid sequence of the light chain variable region of the antibody has at least 90% identity with SEQ ID NO: 27, 28, 29 or 30.

[0114] In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region of the antibody has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with SEQ ID NO: 23, 24, 25 or 26.

[0115] In some embodiments of the first aspect, the amino acid sequence of the light chain variable region of the antibody has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with SEQ ID NO: 27, 28, 29 or 30.

[0116] In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region of the antibody differs from the amino acid sequence shown in SEQ ID NO: 23, 24, 25 or 26 by about 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid substitutions, deletions and / or additions.

[0117] In some embodiments of the first aspect, the amino acid sequence of the light chain variable region of the antibody differs from the amino acid sequence shown in SEQ ID NO: 27, 28, 29 or 30 by about 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid substitutions, deletions and / or additions.

[0118] In some embodiments of the first aspect, the C-terminal or N-terminal region of the amino acid sequence shown in SEQ ID NO: 23, 24, 25 or 26 may also be truncated by about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25 or more amino acids while still maintaining a similar function of the heavy chain variable region of the antibody.

[0119] In some embodiments of the first aspect, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25 or more amino acids can also be added to the C-terminal or N-terminal region of the amino acid sequence shown in SEQ ID NO: 23, 24, 25 or 26, and the resulting amino acid sequence still maintains the function similar to that of the heavy chain variable region of the said antibody.

[0120] In some embodiments of the first aspect, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25 or more amino acids can also be added or deleted from a region other than the C-terminal or N-terminal of the amino acid sequence shown in SEQ ID NO: 23, 24, 25 or 26, as long as the modified amino acid sequence substantially maintains the function similar to that of the heavy chain variable region of the said antibody.

[0121] In some embodiments of the first aspect, the C-terminal or N-terminal region of the amino acid sequence shown in SEQ ID NO: 27, 28, 29 or 30 can also be truncated by about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25 or more amino acids, while still maintaining the function similar to that of the light chain variable region of the said antibody.

[0122] In some embodiments of the first aspect, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25 or more amino acids can also be added to the C-terminal or N-terminal region of the amino acid sequence shown in SEQ ID NO: 27, 28, 29 or 30, and the resulting amino acid sequence still maintains the function similar to that of the light chain variable region of the said antibody.

[0123] In some embodiments of the first aspect, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25 or more amino acids can also be added or deleted from a region other than the C-terminal or N-terminal of the amino acid sequence shown in SEQ ID NO: 27, 28, 29 or 30, as long as the modified amino acid sequence substantially maintains the function similar to that of the light chain variable region of the said antibody.

[0124] In some embodiments of the first aspect, the antibody is a whole antibody, Fab fragment, F(ab’) 2 fragment or single-chain Fv fragment (scFv).

[0125] In some embodiments of the first aspect, the antibody is a fully human antibody.

[0126] In some embodiments of the first aspect, the antibody is a monoclonal antibody.

[0127] In some embodiments of the first aspect, the antibody further comprises a heavy chain constant region selected from the IgG1 subtype, IgG2 subtype, or IgG4 subtype.

[0128] In some embodiments of the first aspect, the heavy chain constant region is of the IgG1 subtype.

[0129] In some embodiments of the first aspect, the heavy chain constant region comprises the Fc segment sequence of the IgG1 subtype heavy chain constant region, and the amino acid sequences at positions 234, 235, and 331 of the Fc segment sequence are F, E, and S, respectively; wherein the amino acid sequence of the antibody constant region is determined according to EU numbering.

[0130] In some embodiments of the first aspect, the heavy chain constant region comprises the Fc segment sequence of the IgG1 subtype heavy chain constant region, and the amino acid sequences at positions 252, 254, and 256 of the Fc segment sequence are Y, T, and E, respectively; wherein the amino acid sequence of the antibody constant region is determined according to EU numbering.

[0131] In some embodiments of the first aspect, the heavy chain constant region comprises the Fc segment sequence of the IgG1 subtype heavy chain constant region, and the amino acid sequences at positions 234, 235, and 331 of the Fc segment sequence are F, E, and S, respectively, and the amino acid sequences at positions 252, 254, and 256 of the Fc segment sequence are Y, T, and E, respectively; wherein the amino acid sequence of the antibody constant region is determined according to EU numbering.

[0132] In some embodiments of the first aspect, the antibody further comprises a light chain constant region selected from the κ subtype or the λ subtype.

[0133] In some embodiments of the first aspect, the antibody binds to TL1A of a primate. In some embodiments, the primate is a human or a monkey (e.g., cynomolgus monkey).

[0134] In some embodiments of the first aspect, the antibody binds to TL1A of a rodent. In some embodiments, the rodent is a murine, such as a rat or a mouse.

[0135] In some embodiments of the first aspect, the antibody binds to recombinant human TL1A (SEQ ID NO: 31).

[0136] In some embodiments of the first aspect, the antibody binds to recombinant monkey (e.g., cynomolgus monkey) TL1A (SEQ ID NO: 32).

[0137] In some embodiments of the first aspect, the antibody binds to recombinant murine TL1A (SEQ ID NO: 33).

[0138] In some embodiments of the first aspect, the antibody binds to recombinant rat TL1A (SEQ ID NO: 34).

[0139] In some embodiments of the first aspect, the antibody inhibits the binding of TL1A to death receptor 3 (DR3).

[0140] In some embodiments of the first aspect, the antibody does not inhibit the binding of TL1A to decoy receptor 3 (DcR3).

[0141] In some embodiments of the first aspect, the antibody inhibits the caspase activation of cells (such as tumor cells, e.g., TF-1 cells) stimulated by TL1A.

[0142] In some embodiments of the first aspect, the antibody inhibits the NF-κB activation of cells (such as 293T-hDR3-NF-κB-RE-luci cells) stimulated by TL1A.

[0143] In some embodiments of the first aspect, the antibody inhibits the ability of TL1A to stimulate PBMC to secrete interferon γ.

[0144] In a second aspect, the present application provides a nucleic acid molecule encoding the antibody described in the first aspect.

[0145] In some embodiments, the nucleic acid molecule is operably linked to a regulatory sequence that can be recognized by a host cell transformed with the vector.

[0146] In a third aspect, the present application provides a pharmaceutical composition comprising the antibody described in the first aspect and a pharmaceutically acceptable excipient, diluent or carrier.

[0147] In some embodiments of the third aspect, the pharmaceutical composition is used for preventing or treating TL1A-related diseases.

[0148] In some embodiments of the third aspect, the TL1A-related diseases are rheumatoid arthritis, inflammatory bowel disease (such as Crohn's disease (CD) and ulcerative colitis (UC)), psoriasis, primary biliary cirrhosis, systemic lupus erythematosus, ankylosing spondylitis and systemic sclerosis-related interstitial lung disease.

[0149] In some embodiments of the third aspect, the pharmaceutical composition may further comprise one or more of the following: lubricants such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying agents; suspending agents; preservatives such as benzoic acid, sorbic acid, and calcium propionate; sweetening agents and / or flavoring agents, etc.

[0150] In some embodiments of the third aspect, the pharmaceutical composition in the present application can be formulated into forms such as tablets, pills, powders, lozenges, elixirs, suspensions, emulsions, solutions, syrups, suppositories, or capsules.

[0151] In some embodiments of the third aspect, the pharmaceutical composition of the present application can be delivered by any physiologically acceptable administration route, including but not limited to: oral administration, parenteral administration, nasal administration, rectal administration, intraperitoneal administration, intravascular injection, subcutaneous administration, transdermal administration, inhalation administration, etc.

[0152] In some embodiments of the third aspect, a pharmaceutical composition for therapeutic use can be formulated in the form of a lyophilized preparation or an aqueous solution for storage by mixing reagents with the desired purity with pharmaceutically acceptable carriers, excipients, etc. as appropriate.

[0153] In a fourth aspect, the present application provides the use of the antibody described in the first aspect and the pharmaceutical composition described in the third aspect in the preparation of a drug for preventing or treating TL1A-related diseases.

[0154] In some embodiments of the fourth aspect, the TL1A-related diseases are rheumatoid arthritis, inflammatory bowel disease (such as Crohn's disease (CD) and ulcerative colitis (UC)), psoriasis, primary biliary cirrhosis, systemic lupus erythematosus, ankylosing spondylitis, and systemic sclerosis-related interstitial lung disease.

[0155] In a fifth aspect, the present application provides a method for preventing or treating TL1A-related diseases, which comprises administering to an individual in need the antibody described in the first aspect, or the pharmaceutical composition described in the third aspect.

[0156] In some embodiments of the fifth aspect, the TL1A-related diseases are rheumatoid arthritis, inflammatory bowel disease (such as Crohn's disease (CD) and ulcerative colitis (UC)), psoriasis, primary biliary cirrhosis, systemic lupus erythematosus, ankylosing spondylitis, and systemic sclerosis-related interstitial lung disease.

[0157] In other aspects, the present application also provides a vector comprising a nucleic acid molecule encoding the antibody of the present invention or its light or heavy chain, a host cell comprising the nucleic acid molecule or the vector, and a method for producing the antibody. In some embodiments, the nucleic acid molecule is operably linked to a regulatory nucleotide sequence that can be recognized by a host cell transformed with the vector. In some embodiments, the method for producing an antibody comprises culturing a host cell to facilitate the expression of the nucleic acid. In some embodiments, the method for producing an antibody further comprises recovering the antibody from the host cell culture medium.

[0158] It should be understood that the above detailed description is only to make those skilled in the art more clearly understand the content of the present application, and is not intended to limit in any way. Those skilled in the art can make various changes and modifications to the embodiments. Examples

[0159] The following examples are for illustrative purposes only and not for the purpose of limiting the scope of the present application.

[0160] Example 1: Preparation of Recombinant Proteins

[0161] A variety of different recombinant proteins are required in the process of preparing and identifying anti-TL1A antibodies, including human TL1A (hTL1A, SEQ ID NO: 31), cynomolgus monkey TL1A (mfTL1A, SEQ ID NO: 32), mouse TL1A (mTL1A, SEQ ID NO: 33), and rat TL1A (rTL1A, SEQ ID NO: 34). Adding the Foldon domain of T4 phage fibritin (Fib Foldon, SEQ ID NO: 35) to the N-terminus of these recombinant proteins helps to maintain the native trimeric conformation of TL1A and enhance stability. At the same time, adding a His tag (His, SEQ ID NO: 36) to the N-terminus of these recombinant proteins is beneficial for the purification and functional identification of the recombinant proteins. When preparing recombinant antibodies, the heavy chain constant region of the antibody can be human IgG1 subtype (CH-IgG1, SEQ ID NO: 37) or various mutants of the defined human IgG1 subtype, such as IgG1m3 (CH-IgG1m3, SEQ ID NO: 38) and IgG1m3-YTE (CH-IgG1m3-YTE, SEQ ID NO: 39); the light chain constant region can be human κ subtype (CK, SEQ ID NO: 40) or human λ subtype (CL, SEQ ID NO: 41).

[0162] According to the amino acid sequences of the recombinant proteins in the Uniprot database, genes of the above various recombinant proteins (including His-tag and Fib Foldon domain) were designed and synthesized. Using conventional molecular biology techniques, the synthesized genes of various recombinant proteins were cloned into appropriate eukaryotic expression vectors (such as pcDNA3.1 from Invitrogen), and then the prepared recombinant protein expression plasmids were transfected into HEK293 cells (such as HEK293F from Invitrogen) using liposomes (such as 293fectin from Invitrogen) or other cationic transfection reagents (such as PEI). The cells were cultured for 3 - 4 days under serum-free suspension culture conditions, and then the culture supernatant was harvested by centrifugation or other methods.

[0163] The recombinant proteins His-fib-hTL1A (SEQ ID NO: 42), His-fib-mfTL1A (SEQ ID NO: 43), His-fib-mTL1A (SEQ ID NO: 44), and His-fib-rTL1A (SEQ ID NO: 45) with His-tag fusion expression were purified in one step from the culture supernatant using a metal chelating affinity chromatography column (such as HisTrap FF from GE). The recombinant antibodies were purified in one step using a Protein A / G affinity chromatography column (such as Mabselect SURE from GE). Then, the storage buffer of the recombinant proteins was replaced with PBS (pH 7.0) or other appropriate buffers using a desalting column (such as Hitrap desaulting from GE). After filtration and sterilization, they were aliquoted and stored at -20 °C for later use.

[0164] Example 2: Preparation and screening of the immune library

[0165] 2.1 Mouse immunization and preparation of the antibody library

[0166] BALB / c mice at 6 - 8 weeks old were taken. Before immunization, blood was collected from the tail vein of the mice to retain the background serum. For the first immunization, the His-fib-hTL1A fusion protein was emulsified with Freund's complete adjuvant, and each mouse was injected with 50 μg of the fusion protein. The booster immunizations were carried out at two-week intervals. For the first and third booster immunizations, the His-fib-hTL1A fusion protein was emulsified with Freund's incomplete adjuvant, and each mouse was injected with 50 μg of the fusion protein. Blood was collected from the tail before injection. For the second and fourth booster immunizations, His-fib-mfTL1A was emulsified with Freund's incomplete adjuvant, and each mouse was injected with 100 μg of the fusion protein. For the sixth immunization, the His-fib-hTL1A recombinant antigen without adjuvant was used as the immunogen, and each mouse was injected with 50 μg of the fusion protein. The mice were sacrificed 3 days after the boost immunization, and spleen cells were collected.

[0167] Mouse spleen lymphocytes were isolated using a mouse lymphocyte separation solution (Beijing DaKeWei Biotechnology Co., Ltd., CAT#DKW33-R0100), and total RNA of lymphocytes was extracted using a total RNA extraction kit (Tiangen Biochemical Technology (Beijing) Co., Ltd., CAT#DP430). Using the extracted total RNA as a template, cDNA of the heavy chain variable region and the light chain variable region was synthesized respectively using a first-strand cDNA synthesis kit (Thermoscientific, CAT#K1621). The reverse transcription primers were gene-specific primers, and the primer pairing regions were located in the antibody heavy chain constant region and the antibody light chain constant region respectively. The specific sequences were PmCGR: TGCATTTGAACTCCTTGCC (SEQ ID NO: 46) and PmCKR: CCATCAATCTTCCACTTGAC (SEQ ID NO: 47). The synthesized cDNA was immediately stored at -70 °C for future use. Then, using the cDNA obtained by reverse transcription as a template, primers were synthesized according to the reference 6 and the VH and VK genes of murine antibodies were amplified by PCR respectively. Then, the single-chain antibody (scFv) gene was constructed using the overlap extension PCR technique. Finally, the prepared murine single-chain antibody gene was cloned into the vector pADSCFV-S (see Chinese Patent Application No. 201510097117.0 7 ), and an scFv library was constructed. The library capacity of this antibody library reached 6.0E+8, and the correct rate was 70%.

[0168] 2.2 Screening of the murine single-chain antibody library

[0169] Using His-fib-hTL1A and His-fib-mfTL1A prepared in Example 1 as antigens, the phage library displaying murine single-chain antibodies constructed above was screened using a solid-phase screening strategy (the experimental protocol refers to Phage Display: A General Experimental Guide, edited by (USA) Clackson, T. and (USA) Lowman, H.B.; translated by Ma Lan et al., Chemical Industry Press, May 2008) 8 Three rounds of screening were carried out in total by means of binding, elution, neutralization, infection, and amplification. Finally, three single-chain antibodies with different sequences, S4D3 (SEQ ID NO: 48), S9H9 (SEQ ID NO: 49), and S7B11 (SEQ ID NO: 50), were obtained.

[0170] Using conventional molecular biology methods, the single-chain antibodies S4D3, S9H9, and S7B11 were respectively prepared into IgG1m3 subtype whole antibodies. At the same time, referring to US Patent No. US20150132311A1 9, synthesize the heavy chain variable region (SEQ ID NO: 51) and the light chain variable region (SEQ ID NO: 52) of RVT-3101; refer to US Patent No. US20210122828A1 10 , synthesize the heavy chain variable region (SEQ ID NO: 53) and the light chain variable region (SEQ ID NO: 54) of PRA023. Prepare RVT-3101 and PRA023 into IgG1m3 subtype full antibodies as positive controls. Prepare DP47 (germline gene antibody, refer to US Patent No. US20160200833A1 11 , DP47 VH, SEQ ID NO: 55; DP47 VK, SEQ ID NO: 56) into IgG1m3 subtype full antibodies as negative controls.

[0171] Example 3: Identification of anti-TL1A murine monoclonal antibodies

[0172] 3.1 Affinity analysis of anti-TL1A murine monoclonal antibodies S4D3, S9H9 and S7B11

[0173] The affinity of mouse anti-human TL1A monoclonal antibody was determined by surface plasmon resonance using Biacore T200. The amino coupling kit (BR-1000-50), human antibody capture kit (BR-1008-39), S series CM5 chip (14100530) and 10×HBS-EP at pH 7.4 (BR100669) and other related reagents and consumables were purchased from GE healthcare. According to the instructions in the kit, the carboxylated CM5 chip surface was activated with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS), and the anti-human IgG (Fc) antibody (capture antibody) was diluted to 25 μg / mL with 10 mM sodium acetate pH 5.0 and then injected at a flow rate of 10 μL / min to achieve a coupling amount of approximately 10,000 response units (RU). After the injection of the capture antibody, 1 M ethanolamine was injected to block unreacted groups. For kinetic measurements, the anti-TL1A monoclonal antibody was diluted to 1 μg / mL and injected at 10 μL / min to ensure that about 200 RU of the antibody was captured by the anti-human Fc antibody. A series of concentration gradients (e.g., 1.23 nM, 3.7 nM, 11.1 nM, 33.3 nM, and 100 nM) were set for His-fib-hTL1A, His-fib-mTL1A, and His-fib-mfTL1A, respectively, and injected from low to high concentrations at a flow rate of 30 μL / min, with an association time of 90 s and a dissociation time of 1200 s. 3 M MgCl was injected at a flow rate of 10 μL / min. 2 The chip surface was regenerated for a total of 30 s. The association rate (K) was calculated by fitting the association and dissociation sensorgrams with a 1:1 binding model using Biacore T200 Evaluation Software version 3.2.1. a ) and dissociation rate (K d ). With the ratio K d / K a Calculate the dissociation equilibrium constant (K D ). The fitting results are shown in Table 1, Table 2 and Table 3.

[0174] Table 1. Affinity constants of TL1A monoclonal antibodies binding to human TL1A (His-fib-hTL1A)

[0175] Antibody <![CDATA[K a (M -1 s -1 ))]]> <![CDATA[K d (s -1 )]]> <![CDATA[K D (M)]]> RVT-3101 6.521E+5 1.001E-4 1.536E-10 PRA023 4.755E+5 1.686E-4 3.546E-10 S4D3 5.323E+5 1.313E-4 2.467E-10 S9H9 1.340E+7 5.012E-4 3.742E-11 S7B11 4.704E+5 1.101E-4 2.341E-10

[0176] Table 2. Affinity constants of TL1A monoclonal antibodies binding to mouse TL1A (His-fib-mTL1A)

[0177] Antibody <![CDATA[K a (M -1 s -1 )]]> <![CDATA[K d (s -1 )]]> <![CDATA[K D (M)]]> RVT-3101 9.039E+5 4.358E-5 4.821E-11 PRA023 NB / / S4D3 NB / / S9H9 6.857E+4 2.551E-3 3.719E-8 S7B11 6.832E+4 3.657E-3 5.353E-8

[0178] NB: No binding detected

[0179] Table 3. Affinity constants of TL1A monoclonal antibodies binding to cynomolgus TL1A (His-fib-mfTL1A)

[0180] Antibody <![CDATA[K a (M -1 s -1 )]]> <![CDATA[K d (s -1 )]]> <![CDATA[K D (M)]]> RVT-3101 1.643E+6 4.515E-5 2.748E-11 PRA023 1.124E+6 9.014E-5 8.023E-11 S4D3 1.290E+6 6.309E-5 4.893E-11 S9H9 2.900E+6 1.749E-4 6.031E-11 S7B11 1.322E+6 6.930E-5 5.202E-11

[0181] 3.2 Inhibitory effect of anti-TL1A murine monoclonal antibodies on caspase activation in TF-1 cells stimulated by TL1A

[0182] TF-1 cells (ATCC, CRL-2003, human erythroleukemia cells) were purchased from ATCC. Cell growth is completely dependent on IL-3 or GM-CSF and is non-responsive to IL-5. These cells express the TL1A receptor DR3, and TL1A can stimulate caspase 3 / 7 activation in these cells. Cycloheximide (CHX) is an inhibitor of eukaryotic protein synthesis and can inhibit cellular protein synthesis. Using RPMI1640 + 5% inactivated FBS as the test medium, TF-1 cells were resuspended in the test medium containing 2 μM Cycloheximide to 2×10 5 cells / mL and plated in 96-well plates at 50 μL / well. The antibody was diluted with the test medium containing 800 ng / mL His-fib-hTL1A. The test antibody started at 40 nM and was serially diluted 1.7-fold, with a total of 10 concentration points. The diluted test antibody was added to TF-1 cells at 50 μL / well, with a total volume of 100 μL / well, and cultured at 37 °C and 5% CO 2 for 5 hours. Caspase 3 / 7 activity was detected using the Caspase-glo 3 / 7 kit (Promega, G8093), and full wavelength scanning was performed using a multimode microplate reader (Molecular Devices, I3X), and fitting analysis was performed based on the chemiluminescence readings. The results ( Figure 1 and Table 4) showed that the anti-TL1A murine monoclonal antibodies S4D3, S9H9, and S7B11 could effectively inhibit caspase activation induced by His-fib-hTL1A, and the activity was comparable to RVT-3101.

[0183] Table 4. Results of the inhibitory effect of anti-TL1A murine monoclonal antibodies on caspase activation in TF-1 cells stimulated by TL1A

[0184] Antibody <![CDATA[IC 50 (nM)]]> RVT-3101 3.129 S4D3 3.047 S9H9 2.686 S7B11 2.686

[0185] 3.3 Anti-TL1A murine monoclonal antibody inhibits NF-κB activation in 293T-hDR3-NF-κB-RE-luci cells stimulated by TL1A

[0186] 293T-NF-κB-RE-luci (product number CS025) was purchased from Yingmao Shengye Biotechnology Co., Ltd. It is a NF-κB reporter gene cell line. Based on this cell line, hDR3 was expressed by liposome transfection, and the 293T-hDR3-NF-κB-RE-luci cell line stably expressing hDR3 was screened for evaluating the activity of anti-TL1A antibody. DMEM + 2% FBS was used as the test medium. The test antibody was diluted with the test medium containing 240 ng / mL His-fib-hTL1A, starting at a concentration of 20 nM, and serially diluted 1.7-fold, with a total of 10 concentration points. 293T-hDR3-NF-κB-RE-luci cells were diluted to 4×10 6 cells / mL. After mixing 50 μL of cells with 50 μL of antibody, they were seeded in a 96-well plate and cultured at 37 °C in 5% CO 2 for 20 hours. The activities of Firefly luciferase and Renilla luciferase were detected successively using the Dual-glo luciferase assay kit (Promega, E2920). A multimode microplate reader (Molecular Devices, I3X) was used for full-wavelength scanning, and the ratio of the Firefly luciferase luminescence signal to the Renilla luciferase luminescence signal was used for fitting analysis. The results ( Figure 2 and Table 5) showed that the anti-TL1A murine monoclonal antibodies S4D3, S9H9, and S7B11 could effectively inhibit the NF-κB signal induced by His-fib-hTL1A, and their activities were comparable to RVT-3101.

[0187] Table 5. Results of the anti-TL1A murine monoclonal antibody inhibiting NF-κB activation in 293T-hDR3-NF-κB-RE-luci cells stimulated by TL1A

[0188] Antibody <![CDATA[IC 50 (nM)]]> S9H9 1.284 S7B11 1.346 RVT-3101 1.278 S4D3 1.128

[0189] Example 4: Screening of a fully human Fab recombinant library

[0190] Using the recombinant His-fib-hTL1A and His-fib-mfTL1A prepared in Example 1 as antigens, a fully human Fab phage library was screened using a solid-phase screening strategy (for the experimental protocol, refer to Phage Display: A General Experimental Guide, edited by (US) Clackson, T. and (US) Lowman, H.B.; translated by Ma Lan et al., Chemical Industry Press, May 2008). A total of 3 rounds of screening were carried out by means of binding, elution, neutralization, infection, and amplification, and finally a monoclonal antibody H3F1+L28E1 that specifically binds to human, cynomolgus monkey, mouse, and rat TL1A was obtained (the amino acid sequence of H3F1VH is shown in SEQ ID NO: 26; the amino acid sequence of L28E1VK is shown in SEQ ID NO: 30).

[0191] Using conventional molecular biology techniques, H3F1+L28E1 was prepared into a full antibody of the IgG1m3 subtype.

[0192] Example 5: Identification of a fully human anti-TL1A monoclonal antibody

[0193] 5.1 Affinity analysis of the anti-TL1A monoclonal antibody H3F1+L28E1

[0194] Referring to Example 3.1, the Biacore T200 was used to analyze the affinity of the TL1A monoclonal antibody H3F1+L28E1 for binding to TL1A of different species. The results are shown in Tables 6, 7, 8, and 9.

[0195] Table 6. Affinity constant of the anti-TL1A monoclonal antibody H3F1+L28E1 for binding to human TL1A (His-fib-hTL1A)

[0196] Antibody <![CDATA[K a (M -1 s -1 )]]> <![CDATA[K d (s -1 )]]> <![CDATA[K D (M)]]> RVT-3101 7.063E+5 1.016E-4 1.438E-10 PRA023 4.755E+5 1.686E-4 3.546E-10 H3F1+L28E1 5.381E+6 1.562E-4 2.093E-11

[0197] Table 7. Affinity constant of the anti-TL1A monoclonal antibody H3F1+L28E1 for binding to mouse TL1A (His-fib-mTL1A)

[0198] Antibody <![CDATA[K a (M -1 s -1 )]]> <![CDATA[K d (s -1 )]]> <![CDATA[K D (M) <!-- 13 -->]]> RVT-3101 7.635E+5 5.92E-5 7.753E-11 PRA023 NB / / H3F1+L28E1 9.643E+5 1.831E-3 1.898E-9

[0199] NB: Binding not detected

[0200] Table 8. Affinity constant of the anti-TL1A monoclonal antibody H3F1+L28E1 for binding to rat TL1A (His-fib-rTL1A)

[0201] Antibody <![CDATA[K a (M -1 s -1 )]]> <![CDATA[K d (s -1 )]]> <![CDATA[K D (M)]]> RVT-3101 9.778E+5 4.243E-4 4.34E-10 PRA023 NB / / H3F1+L28E1 8.037E+6 1.170E-3 1.456E-10

[0202] NB: Binding not detected

[0203] Table 9. Affinity Constant of Anti-TL1A Monoclonal Antibody H3F1+L28E1 Binding to Monkey TL1A (His-fib-mfTL1A)

[0204] Antibody <![CDATA[K a (M -1 s -1 )]]> <![CDATA[K d (s -1 )]]> <![CDATA[K D (M)]]> RVT-3101 1.340E+6 6.700E-5 4.998E-11 PRA023 1.124E+6 9.014E-5 8.023E-11 H3F1+L28E1 4.191E+6 1.330E-4 3.173E-11

[0205] 5.2 H3F1+L2831 Inhibits Caspase Activation in TF-1 Cells Stimulated by TL1A

[0206] Referring to Example 3.2, evaluate the inhibitory effect of H3F1+L2831 on caspase activation in TF-1 cells stimulated by TL1A. Dilute the antibody with the test medium containing 800 ng / mL His-fib-hTL1A. All test antibodies start at 40 nM and are serially diluted 1.7-fold, with a total of 10 concentration points. The results ( Figure 3 and Table 10) show that H3F1+L28E1 can effectively inhibit caspase signaling induced by His-fib-TL1A, and its activity is superior to that of the RVT-3101 and PRA023 control antibodies.

[0207] Table 10. Results of the Inhibitory Effect of H3F1+L2831 on Caspase Activation in TF-1 Cells Stimulated by TL1A

[0208] Antibody <![CDATA[IC 50 (nM)]]> RVT-3101 3.464 PRA023 6.523 H3F1+L28E1 1.734

[0209] 5.3 H3F1+L28E1 Inhibits NF-κB Activation in 293T-hDR3-NF-κB-RE-luci Cells Stimulated by TL1A

[0210] Referring to Example 3.3, evaluate the inhibitory effect of H3F1+L28E1 on NF-κB activation in 293T-hDR3-NF-κB-RE-luci cells stimulated by TL1A. Dilute the test antibody with the test medium containing 240 ng / mL His-fib-hTL1A. The starting concentration is 20 nM, and it is serially diluted 1.7-fold, with a total of 10 concentration points. The results ( Figure 4 and Table 11) show that H3F1+L28E1 can effectively inhibit NF-κB signaling stimulated by His-fib-hTL1A, and its activity is superior to that of RVT-3101 and PRA023.

[0211] Table 11. Results of the Inhibitory Effect of H3F1+L28E1 on NF-κB Activation in 293T-hDR3-NF-κB-RE-luci Cells Stimulated by TL1A

[0212] Antibody <![CDATA[IC 50 (nM)]]> RVT-3101 1.082 PRA023 2.813 H3F1+L28E1 0.574

[0213] 5.4H3F1 + L28E1 inhibits the ability of TL1A to stimulate PBMC to secrete interferon γ

[0214] The TL1A receptor DR3 is expressed on the surface of NK cells or activated T cells. With the help of co-stimulatory factors such as IL-12 / IL-18, after TL1A binds to NK cells or activated T cells, it can activate the downstream signaling pathway, ultimately leading to the secretion of interferon γ. By detecting the content of interferon γ in the cell supernatant, the inhibitory effect of anti-TL1A antibodies on TL1A can be evaluated.

[0215] Collect blood (50 mL) from normal volunteers, which is provided by the inventor and his colleagues as volunteers, and all volunteers have signed informed consent forms. The inclusion criteria for volunteers are:

[0216] 1. Age greater than 18 years old;

[0217] 2. No HIV or HBV infection;

[0218] 3. Normal blood routine test;

[0219] 4. Not pregnant or lactating women.

[0220] Use Ficoll density gradient centrifugation to isolate human peripheral blood mononuclear cells (PBMC) from healthy human peripheral blood. Use RPMI 1640 + 5% inactivated FBS as the test medium, and resuspend PBMC in the test medium containing 0.5 ng / mL IL-12 and 2 ng / mL IL-18 to 6×10 5 cells / mL, and plate 100 μL / well in a 96-well plate, and incubate statically at 37°C and 5% CO 2 for 3 hours. After 3 hours, dilute the antibody with the test medium containing 2.4 μg / mL His-fib-hTL1A. The test antibody starts at 200 nM and is serially diluted 2.3-fold, with a total of 8 concentration points. The diluted antibody is added to PBMC at 100 μL / well, with a total volume of 200 μL / well, and incubated at 37°C and 5% CO 2 for 20 hours. Use a human IFN-γ pre-coated ELISA kit (Dayou, 1110002) to detect the content of interferon γ in the cell supernatant. Read the value at OD450nm with an enzyme-linked immunosorbent assay (800TS, Biotek), draw a standard curve of concentration and absorbance with the interferon γ standard product provided in the detection kit, and obtain the content of interferon γ in the supernatant by conversion and perform fitting analysis. The results ( Figure 5 and Table 12) show that H3F1 + L28E1 can effectively inhibit His-fib-hTL1A from stimulating PBMC to secrete interferon γ, and its activity is superior to RVT-3101 and PRA023.

[0221] Table 12. Ability of H3F1+L28E1 to inhibit interferon-γ secretion by PBMC stimulated with TL1A

[0222] Antibody <![CDATA[IC 50 (nM)]]> RVT-3101 13.61 PRA023 6.545 H3F1+L28E1 5.232

[0223] Example 6: Anti-TL1A monoclonal antibody selectively neutralizes the functional receptor DR3

[0224] The human DR3 (Recombinant Human DR3 / TNFRSF25 Fc Chimera Protein, CF, R&D SYSTEMS, 943-D3-050) was biotinylated using a biotinylation kit (EZ-Link TM Sulfo-NHS-Biotin, No-Weigh TM Format, ThermoFisher, A39256). His-fib-hTL1A was coated on a 96-well ELISA plate at 4 μg / mL, 100 μL / well, and incubated overnight at 4°C. The plate was blocked with a blocking solution (3% skim milk-PBST) at 37°C for 1 hour. The anti-human TL1A monoclonal antibodies (RVT-3101 and H3F1+L28E1) were serially diluted with biotinylated human DR3 at 10 μg / mL, starting from a concentration of 200 μg / mL, with a 3-fold serial dilution for a total of 8 concentration gradients, and 100 μL / well was added to the blocked 96-well ELISA plate and incubated at 37°C for 1 hour. The ELISA plate was washed with PBST, and then streptavidin conjugated to horseradish peroxidase (Streptavidin / HRP, Beijing Boao Shen Biotechnology Co., Ltd., bs-0437P-HRP) was added and incubated at 37°C for 1 hour. The ELISA plate was washed with PBST, OPD substrate chromogenic solution was added, and the color development was terminated with 1 M H 2 SO 4 after 5 - 10 minutes, and the optical density value at a single wavelength of 490 nm was measured using a microplate reader. The results of ELISA analysis are as Figure 6 shown: RVT-3101 and H3F1+L28E1 can block the binding of human DR3 to human TL1A.

[0225] Coat a 96-well ELISA plate with human DcR3 (Recombinant Human DcR3 / TNFRSF6B Fc Chimera Protein, CF, R&D SYSTEMS, 142-DC-100) at 3 μg / mL, 100 μL / well, and incubate overnight at 4°C. Block with a blocking solution (3% skim milk-PBST) at 37°C for 1 hour. Gradient dilute anti-human TL1A monoclonal antibodies (RVT-3101, PRA023, S4D3, S9H9, S7B11, and H3F1+L28E1) with 3 μg / mL of His-fib-hTL1A, starting from a concentration of 100 μg / mL, with a 3-fold serial dilution, for a total of 11 concentration gradients, and add 100 μL / well to the blocked 96-well ELISA plate. Incubate at 37°C for 1 hour. Wash the ELISA plate with PBST, then add an HRP-labeled anti-His tag mouse monoclonal antibody (ComWin Biotech, cw0285M), and incubate at 37°C for 1 hour. Wash the ELISA plate with PBST, add the OPD substrate chromogenic solution, and after 5 - 10 minutes, use 1 M H 2 SO 4 to terminate the color development, and measure the optical density value at a single wavelength of 490 nm using an enzyme-linked immunosorbent assay reader. The results of the ELISA analysis are as Figure 7 shown: RVT-3101 and PRA023 can block the binding of human DcR3 to human TL1A, while S4D3, S9H9, S7B11, and H3F1+L28E1 do not block the binding of human DcR3 to human TL1A.

[0226] As a common decoy receptor for TNF family cytokines (Fas-L, LIGHT, and TL1A), DcR3 has a natural antagonistic function in vivo. S4D3, S9H9, S7B11, and H3F1+L28E1 can neutralize DR3 but not DcR3, indicating that these antibodies do not disrupt the natural antagonistic activity of DcR3, help maintain the homeostatic balance of DcR3 in vivo, and have better safety.

[0227] Example 7: Effect of anti-TL1A monoclonal antibody on DNBS-induced acute ulcerative colitis in rats

[0228] Male Wistar rats with no history of drug administration were purchased from Shanghai Slack Experimental Animal Breeding Co., Ltd. and were allowed to acclimatize to the animal house environment 3 days in advance. Before the DNBS-induced model was established, the rats were fasted for 40 hours, and during the fasting period, 5% glucose injection (10 mL / kg) was subcutaneously injected as an energy supplement during fasting. The DNBS powder was dissolved in 30% ethanol to a final concentration of 60 mg / mL. On day 0, the fasted rats were anesthetized with Zoletil (intraperitoneal injection, 25 - 50 mg / kg) and xylazine (intraperitoneal injection, 5 - 10 mg / kg). The specific animal grouping and dosing regimens are shown in Table 13. Among groups G2 - G4, a soft tube was inserted from the anus into the colon, and DNBS enema was used to induce colitis in rats. In group G1, enema was performed in the same way with 30% ethanol. The isotype control was DP47 antibody.

[0229] Table 13. Grouping and dosing regimens

[0230]

[0231] During the experiment, the fecal traits of the experimental animals were scored daily (0 = normal, 1 = moist / sticky, 2 = soft, 3 = liquid). The body weights of the experimental animals were measured and recorded daily. On day 6, all experimental animals were euthanized by asphyxiation with excessive carbon dioxide. Then, the abdominal cavity was opened, the colon was removed, longitudinally dissected, rinsed thoroughly, and the ulcer surface of the colon was observed. The colon length, weight, and ulcer area were recorded, and a macroscopic damage score of the colon was made. Results ( Figure 8 and Figure 9 ) showed that the fecal traits and intestinal injuries of the experimental animals in group G4 (H3F1 + L28E1) were significantly improved compared with those in group G2 (model - isotype control group).

[0232] All patents, patent application publications, and non - patent literature mentioned and / or listed in this application are hereby incorporated by reference in their entirety. The above - described exemplary embodiments of the inventions of this application have been described. However, without departing from the essence and scope of this application, those skilled in the art can modify or improve the exemplary embodiments described in this application, and the resulting variant or equivalent embodiments also fall within the scope of this application.

[0233] Sequence information

[0234] SEQ ID NO:1

[0235] KYDIN

[0236] SEQ ID NO:2

[0237] WIFPGDGRTDYNEKFKG

[0238] SEQ ID NO:3

[0239] YGYALDY

[0240] SEQ ID NO:4

[0241] RSSQNIVHSNGDTYLE

[0242] SEQ ID NO:5

[0243] KVSNRFS

[0244] SEQ ID NO:6

[0245] FQGSHFPYT

[0246] SEQ ID NO:7

[0247] NYWLG

[0248] SEQ ID NO:8

[0249] DIHPGRGNIFYNEKFKG

[0250] SEQ ID NO:9

[0251] GYDTFDY

[0252] SEQ ID NO:10

[0253] KASQDVSTAVA

[0254] SEQ ID NO:11

[0255] SASYRYT

[0256] SEQ ID NO:12

[0257] QQHYSTPWT

[0258] SEQ ID NO:13

[0259] DIHPGRGNIYYNEKFKG

[0260] SEQ ID NO:14

[0261] RASKSISKYLA

[0262] SEQ ID NO:15

[0263] SGSTLQS

[0264] SEQ ID NO:16

[0265] QQHNEYPYT

[0266] SEQ ID NO:17

[0267] SYDIN

[0268] SEQ ID NO:18

[0269] WLNPNSGNTGYAQKFQG

[0270] SEQ ID NO:19

[0271] EIPESAAIEY

[0272] SEQ ID NO:20

[0273] TSSSSDIGAGLGVH

[0274] SEQ ID NO:21

[0275] GYYNRPS

[0276] SEQ ID NO:22

[0277] QSYDASLTGI

[0278] SEQ ID NO:23

[0279] QVQLKESGSDLATPGASVKLSCKVSGYTFTKYDINWVRQRPEQGLEWIGWIFPGDGRTDYNEKFKGKATLTIDTSSSTAYMQLSRLTSEDSAVYFCARYGYALDYWGQGTSVTVSS

[0280] SEQ ID NO:24

[0281] QVQLKQSGAELVRPGTSVKISCKASGYAFTNYWLGWIKQRPGHGLEWIGDIHPGRGNIFYNEKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYFCAEGYDTFDYWGQGTTLTVSA

[0282] SEQ ID NO:25

[0283] QVKLQQSGGELVRPGTSVKISCKASGYAFTNYWLGWIKQRPGHGLEWIGDIHPGRGNIYYNEKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYFCAEGYDTFDYWGQGTTLTVSS

[0284] SEQ ID NO:26

[0285] QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYDINWVRQAPGQGLEWMGWLNPNSGNTGYAQKFQGRVTMTADRSTSTAYMELSSLRSEDTAVYYCAREIPESAAIEYWGQGTLVTVSS

[0286] SEQ ID NO:27

[0287] DVVMTQTPLSLPVSLGDQASISCRSSQNIVHSNGDTYLEWFLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHFPYTFGGGTKLEMK

[0288] SEQ ID NO:28

[0289] DIVMTQSHKFMSTSVGDRVSITCKASQDVSTAVAWYQQKPGQSPKLLIYSASYRYTGVPDRFTGSGSGTDFTFTISSVQAEDLAVYYCQQHYSTPWTFGGGTKLEMK

[0290] SEQ ID NO:29

[0291] DIVMTQSPSYLAASPGETITINCRASKSISKYLAWYQEKPGKTNKLLIYSGSTLQSGIPSRFSGSGSGTDFTLTISSLEPEDFAMYYCQQHNEYPYTFGGGTKLEIK

[0292] SEQ ID NO:30

[0293] QSVLTQPPSVSGAPGQRVTISCTSSSSDIGAGLGVHWYQQLPGTAPKLLIEGYYNRPSGVPDRFSGSKSGTSASLTITGLLPEDEGDYYCQSYDASLTGIFGGGTKLTVL

[0294] SEQ ID NO:31

[0295] LKGQEFAPSHQQVYAPLRADGDKPRAHLTVVRQTPTQHFKNQFPALHWEHELGLAFTKNRMNYTNKFLLIPESGDYFIYSQVTFRGMTSECSEIRQAGRPNKPDSITVVITKVTDSYPEPTQLLMGTKSVCEVGSNWFQPIYLGAMFSLQEGDKLMVNVSDISLVDYTKEDKTFFGAFLL

[0296] SEQ ID NO:32

[0297] LKGQEFAPSHQQVYAPLRADGDKPRAHLTVVRQTPTQHLKNQFPALHWEHELGLAFTKNRMNYTNKFLLIPESGDYFVYSQVTFRGMTSECSEIRQAGRPNKPDSITVVITKVTDSYPEPTQLLMGTKSVCEVGSNWFQPIYLGAMFSLQEGDKLMVNVSDISLVDYTKEDKTFFGAFLL

[0298] SEQ ID NO:33

[0299] ITEERSEPSPQQVYSPPRGKPRAHLTIKKQTPAPHLKNQLSALHWEHDLGMAFTKNGMKYINKSLVIPESGDYFIYSQITFRGTTSVCGDISRGRRPNKPDSITMVITKVADSYPEPARLLTGSKSVCEISNNWFQSLYLGATFSLEEGDRLMVNVSDISLVDYTKEDKTFFGAFLL

[0300] SEQ ID NO:34

[0301] VTEERSAPSAQPVYTPSRDKPKAHLTIMRQTPVPHLKNELAALHWENNLGMAFTKNRMNYTNKFLVIPESGDYFIYSQITFRGTTSECGDISRVRRPKKPDSITVVITKVADSYPEPAHLLTGTKSVCEISSNWFQPIYLGAMFSLEEGDRLMVNVSDISLVDYTKEDKTFFGAFLI

[0302] SEQ ID NO:35

[0303] GYIPEAPRDGQAYVRKDGEWVLLSTFL

[0304] SEQ ID NO:36

[0305] HHHHHH

[0306] SEQ ID NO:37

[0307] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0308] SEQ ID NO:38

[0309] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0310] SEQ ID NO:39

[0311] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0312] SEQ ID NO:40

[0313] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0314] SEQ ID NO:41

[0315] GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS

[0316] SEQ ID NO:42

[0317] HHHHHHGGGGSGYIPEAPRDGQAYVRKDGEWVLLSTFLGGGGSLKGQEFAPSHQQVYAPLRADGDKPRAHLTVVRQTPTQHFKNQFPALHWEHELGLAFTKNRMNYTNKFLLIPESGDYFIYSQVTFRGMTSECSEIRQAGRPNKPDSITVVITKVTDSYPEPTQLLMGTKSVCEVGSN

[0318] WFQPIYLGAMFSLQEGDKLMVNVSDISLVDYTKEDKTFFGAFLL

[0319] SEQ ID NO:43

[0320] HHHHHHGGGGSGYIPEAPRDGQAYVRKDGEWVLLSTFLGGGGSLKGQEFAPSHQQVYAPLRADGDKPRAHLTVVRQTPTQHLKNQFPALHWEHELGLAFTKNRMNYTNKFLLIPESGDYFVYSQVTFRGMTSECSEIRQAGRPNKPDSITVVITKVTDSYPEPTQLLMGTKSVCEVGSNWFQPIYLGAMFSLQEGDKLMVNVSDISLVDYTKEDKTFFGAFLL

[0321] SEQ ID NO:44

[0322] HHHHHHGGGGSGYIPEAPRDGQAYVRKDGEWVLLSTFLGGGGSITEERSEPSPQQVYSPPRGKPRAHLTIKKQTPAPHLKNQLSALHWEHDLGMAFTKNGMKYINKSLVIPESGDYFIYSQITFRGTTSVCGDISRGRRPNKPDSITMVITKVADSYPEPARLLTGSKSVCEISNNWFQSLYLGATFSLEEGDRLMVNVSDISLVDYTKEDKTFFGAFLL

[0323] SEQ ID NO:45

[0324] HHHHHHGGGGSGYIPEAPRDGQAYVRKDGEWVLLSTFLGGGGSVTEERSAPSAQPVYTPSRDKPKAHLTIMRQTPVPHLKNELAALHWENNLGMAFTKNRMNYTNKFLVIPESGDYFIYSQITFRGTTSECGDISRVRRPKKPDSITVVITKVADSYPEPAHLLTGTKSVCEISSNWFQPIYLGAMFSLEEGDRLMVNVSDISLVDYTKEDKTFFGAFLI

[0325] SEQ ID NO:46

[0326] TGCATTTGAACTCCTTGCC

[0327] SEQ ID NO:47

[0328] CCATCAATCTTCCACTTGAC

[0329] SEQ ID NO:48

[0330] QVQLKESGSDLATPGASVKLSCKVSGYTFTKYDINWVRQRPEQGLEWIGWIFPGDGRTDYNEKFKGKATLTIDTSSSTAYMQLSRLTSEDSAVYFCARYGYALDYWGQGTSVTVSSGGGGSGGGGSGGGGSDVVMTQTPLSLPVSLGDQASISCRSSQNIVHSNGDTYLEWFLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHFPYTFGGGTKLEMK

[0331] SEQ ID NO:49

[0332] QVQLKQSGAELVRPGTSVKISCKASGYAFTNYWLGWIKQRPGHGLEWIGDIHPGRGNIFYNEKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYFCAEGYDTFDYWGQGTTLTVSAGGGGSGGGGSGGGGSDIVMTQSHKFMSTSVGDRVSITCKASQDVSTAVAWYQQKPGQSPKLLIYSASYRYTGVPDRFTGSGSGTDFTFTISSVQAEDLAVYYCQQHYSTPWTFGGGTKLEMK

[0333] SEQ ID NO:50

[0334] QVKLQQSGGELVRPGTSVKISCKASGYAFTNYWLGWIKQRPGHGLEWIGDIHPGRGNIYYNEKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYFCAEGYDTFDYWGQGTTLTVSSGGGGSGGGGSGGGGSDIVMTQSPSYLAASPGETITINCRASKSISKYLAWYQEKPGKTNKLLIYSGSTLQSGIPSRFSGSGSGTDFTLTISSLEPEDFAMYYCQQHNEYPYTFGGGTKLEIK

[0335] SEQ ID NO:51

[0336] QVQLVQSGAEVKKPGASVKVSCKASGYDFTYYGISWVRQAPGQGLEWMGWISTYNGNTHYARMLQGRVTMTTDTSTRTAYMELRSLRSDDTAVYYCARENYYGSGAYRGGMDVWGQGTTVTVSS

[0337] SEQ ID NO:52

[0338] EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPWTFGQGTKVEIK

[0339] SEQ ID NO:53QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVKQRPGQGLEWMGRIDPASGHTKYDPKFQVRVTITRDTSTSTVYLELSSLRSEDTAVYYCARSGGLPDVWGQGTTVTVSS

[0340] SEQ ID NO:54

[0341] EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRPLIYATSNLASGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQWEGNPRTFGGGTKLEIK

[0342] SEQ ID NO:55

[0343] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGSGFDYWGQGTLVTVSS

[0344] SEQ ID NO:56

[0345] EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPLTFGQGTKVEIK

[0346] References

[0347] 1. Xu WD, Li R, Huang AF. Role of TL1A in Inflammatory Autoimmune Diseases: A Comprehensive Review. Front Immunol. 2022 Jul 14; 13: 891328.

[0348] 2. Migone TS, Zhang J, Luo X, Zhuang L, Chen C, Hu B, Hong JS, Perry JW, Chen SF, Zhou JX, Cho YH, Ullrich S, Kanakaraj P, Carrell J, Boyd E, Olsen HS, Hu G, Pukac L, Liu D, Ni J, Kim S, Gentz R, Feng P, Moore PA, Ruben SM, Wei P. TL1A is a TNF-like ligand for DR3 and TR6 / DcR3 and functions as a T cell costimulator. Immunity. 2002 Mar; 16(3): 479-92.

[0349] 3. Valatas V, Kolios G, Bamias G. TL1A (TNFSF15) and DR3 (TNFRSF25): A Co-stimulatory System of Cytokines With Diverse Functions in Gut Mucosal Immunity. Front Immunol. 2019 Mar 27; 10: 583.

[0350] 4. Ramos GP, Papadakis KA. Mechanisms of Disease: Inflammatory Bowel Diseases. Mayo Clin Proc. 2019 Jan; 94(1): 155-165.

[0351] 5. Li N, Ye M. Advances in biological agents in the treatment of inflammatory bowel disease[J]. Yixue Xinzhi Zazhi, 2022, 32(4): 310-320.

[0352] 6. Krebber A, Bornhauser S, Burmester J, Honegger A, Willuda J, Bosshard HR, Plückthun A. Reliable cloning of functional antibody variable domains from hybridomas and spleen cell repertoires employing a reengineered phage display system. J Immunol Methods. 1997 Feb 14;201(1):35 - 55.

[0353] 7. CN201510097117.0.

[0354] 8. Phage Display: A General Laboratory Guide / Edited by (US) Clackson, T., (US) Lowman, H.B.; Translated by Ma Lan, etc. Chemical Industry Press, 2008.5.

[0355] 9. US20150132311A1

[0356] 10. US20210122828A1

[0357] 11. US20160200833A1.

Claims

1. An antibody that binds to tumor necrosis factor-like ligand 1A (TL1A), comprising a heavy chain variable region containing the amino acid sequences of HCDR1, HCDR2, and HCDR3, and a light chain variable region containing the amino acid sequences of LCDR1, LCDR2, and LCDR3, wherein the amino acid sequence of said HCDR1 is as shown in SEQ ID NO: 1, the amino acid sequence of said HCDR2 is as shown in SEQ ID NO: 2, the amino acid sequence of said HCDR3 is as shown in SEQ ID NO: 3, the amino acid sequence of said LCDR1 is as shown in SEQ ID NO: 4, the amino acid sequence of said LCDR2 is as shown in SEQ ID NO: 5, and the amino acid sequence of said LCDR3 is as shown in SEQ ID NO: 6; or the amino acid sequence of said HCDR1 is as shown in SEQ ID NO: 7, the amino acid sequence of said HCDR2 is as shown in SEQ ID NO: 8, the amino acid sequence of said HCDR3 is as shown in SEQ ID NO: 9, the amino acid sequence of said LCDR1 is as shown in SEQ ID NO: 10, the amino acid sequence of said LCDR2 is as shown in SEQ ID NO: 11, and the amino acid sequence of said LCDR3 is as shown in SEQ ID NO: 12; or the amino acid sequence of said HCDR1 is as shown in SEQ ID NO: 7, the amino acid sequence of said HCDR2 is as shown in SEQ ID NO: 13, the amino acid sequence of said HCDR3 is as shown in SEQ ID NO: 9, the amino acid sequence of said LCDR1 is as shown in SEQ ID NO: 14, the amino acid sequence of said LCDR2 is as shown in SEQ ID NO: 15, and the amino acid sequence of said LCDR3 is as shown in SEQ ID NO: 16; or the amino acid sequence of said HCDR1 is as shown in SEQ ID NO: 17, the amino acid sequence of said HCDR2 is as shown in SEQ ID NO: 18, the amino acid sequence of said HCDR3 is as shown in SEQ ID NO: 19, the amino acid sequence of said LCDR1 is as shown in SEQ ID NO: 20, the amino acid sequence of said LCDR2 is as shown in SEQ ID NO: 21, and the amino acid sequence of said LCDR3 is as shown in SEQ ID NO: 22; wherein, the HCDR and LCDR amino acid sequences are defined according to Kabat.

2. The antibody according to claim 1, wherein the amino acid sequence of the heavy chain variable region of said antibody is as shown in SEQ ID NO: 23, 24, 25, or 26.

3. The antibody according to claim 1, wherein the amino acid sequence of the light chain variable region of said antibody is as shown in SEQ ID NO: 27, 28, 29, or 30.

4. The antibody according to any one of claims 1-3, wherein The amino acid sequence of the variable region of the heavy chain of the antibody is as shown in SEQ ID NO: 23, and the amino acid sequence of the variable region of the light chain of the antibody is as shown in SEQ ID NO: 27; or The amino acid sequence of the variable region of the heavy chain of the antibody is as shown in SEQ ID NO: 24, and the amino acid sequence of the variable region of the light chain of the antibody is as shown in SEQ ID NO: 28; or The amino acid sequence of the variable region of the heavy chain of the antibody is as shown in SEQ ID NO: 25, and the amino acid sequence of the variable region of the light chain of the antibody is as shown in SEQ ID NO: 29; or The amino acid sequence of the variable region of the heavy chain of the antibody is as shown in SEQ ID NO: 26, and the amino acid sequence of the variable region of the light chain of the antibody is as shown in SEQ ID NO:

30.

5. The antibody according to any one of claims 1-4, wherein the amino acid sequence of the variable region of the heavy chain of the antibody has at least 90% identity with SEQ ID NO: 23, 24, 25 or 26, and the amino acid sequence of the variable region of the light chain of the antibody has at least 90% identity with SEQ ID NO: 27, 28, 29 or 30.

6. The antibody according to any one of claims 1-5, wherein The antibody is a whole antibody, Fab fragment, F(ab’) 2 fragment or single-chain Fv fragment (scFv), preferably, the antibody is a fully human antibody; and / or The antibody is a monoclonal antibody; and / or The antibody further comprises a heavy chain constant region selected from the IgG1 subtype, IgG2 subtype or IgG4 subtype; preferably, the heavy chain constant region is the IgG1 subtype; more preferably, the heavy chain constant region comprises the Fc segment sequence of the IgG1 subtype heavy chain constant region, and the amino acid sequences at positions 234, 235, and 331 of the Fc segment sequence are F, E, and S, respectively, and / or the amino acid sequences at positions 252, 254, and 256 of the Fc segment sequence are Y, T, and E, respectively; wherein the amino acid sequence of the constant region of the antibody is determined according to EUnumbering; and / or The antibody further comprises a light chain constant region selected from the κ subtype or the λ subtype.

7. The antibody according to any one of claims 1-6, wherein The antibody binds to TL1A of primates and / or rodents; preferably, the primate is a human or a monkey (e.g., cynomolgus monkey), and / or the rodent is a murine (e.g., rat or mouse); and / or The antibody inhibits the binding of TL1A to death receptor 3 (DR3); and / or The antibody does not inhibit the binding of TL1A to decoy receptor 3 (DcR3); and / or The antibody inhibits the caspase activation of cells stimulated by TL1A; and / or The antibody inhibits the NF-κB activation of cells stimulated by TL1A; and / or The antibody inhibits the ability of TL1A to stimulate PBMC to secrete interferon γ.

8. A nucleic acid molecule encoding the antibody according to any one of claims 1-7.

9. A pharmaceutical composition comprising the antibody according to any one of claims 1-7 and a pharmaceutically acceptable excipient, diluent or carrier. Use of the antibody according to any one of claims 1-7, or the pharmaceutical composition according to claim 9, in the preparation of a medicament for preventing or treating TL1A-related diseases; preferably, the TL1A-related diseases are rheumatoid arthritis, inflammatory bowel disease (such as Crohn's disease (CD) and ulcerative colitis (UC)), psoriasis, primary biliary cirrhosis, systemic lupus erythematosus, ankylosing spondylitis, and interstitial lung disease related to systemic sclerosis.

Images