Tl1a antibodies and uses thereof

Abstract

This disclosure pertains to the field of biomedicine, specifically relating to TL1A antibodies and their applications. The antibody or its antigen-binding fragment disclosed herein can bind to TL1A, block the binding of TL1A to DR3, inhibit TL1A-induced TF-1 cell apoptosis, inhibit TL1A-induced NF-κB signaling pathway activation, and inhibit TL1A-induced IFN-γ expression in blood cells.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, specifically to TL1A antibodies and their applications. Background Technology

[0002] Tumor necrosis factor (TNF)-like ligand 1A (TL1A) is a member of the TNF family of cytokines, also known as TNFSF15. TL1A is the only known ligand for receptor death receptor 3 (DR3) (also known as TNFRSF25), and only the trimer form of TL1A has the ability to bind to and activate DR3. TL1A can be produced by umbilical vein endothelial cells (HUVECs), monocytes, macrophages, dendritic cells (DCs), T cells, chondrocytes, and synovial fibroblasts, which are the sources of both soluble and membrane-bound forms of TL1A. TL1A participates in intrinsic immunity by influencing monocyte lineages, DCs, natural killer cells (NK cells), and innate lymphocytes (ILCs) to regulate the immune environment. Soluble TL1A (sTL1A) exerts various effects on adaptive immune cells by binding to DR3, affecting the activation, proliferation, and differentiation of different immune cells, such as helper T cells, regulatory T cells, B cells, and the production of cytokines.

[0003] Studies in DR3 / TL1A knockout mice or mice treated with anti-TL1A antibodies have shown that the DR3-TL1A signaling pathway plays a crucial role in various autoimmune disease models, such as IBD, asthma, multiple sclerosis, and arthritis. Furthermore, extensive human genetic data indicate that TL1A is closely associated with the pathophysiology of inflammatory bowel diseases (IBD), such as ulcerative colitis (UC) and Crohn's disease (CD). Numerous genome-wide association studies have found that several polymorphisms of the TL1A gene are associated with ulcerative colitis and Crohn's disease in patient populations of Japanese, European, and Asian descent. Moreover, in the intestinal tissue of inflamed IBD patients, the expression levels of both DR3 and TL1A are significantly higher than in adjacent non-inflammatory sites. It has been reported that in mouse IBD models, blocking TL1A with antibodies can prevent or reduce the degree of inflammation in the mouse intestinal tissue. TL1A can also act on fibroblasts, promoting tissue fibrosis. Currently, numerous studies have revealed the role of TL1A in intestinal fibrosis, liver fibrosis, and pulmonary fibrosis, making it a promising therapeutic target for fibrosis.

[0004] While the exact causes of inflammatory bowel diseases (such as CD and UC) remain unclear, inhibiting disease-associated pro-inflammatory cytokines or adhesion molecules has been shown to offer some therapeutic benefits. However, despite the availability of therapeutic drugs, most CD patients will eventually require surgery, and repeated resections of diseased intestinal tissue over time can lead to short bowel syndrome, ultimately requiring lifelong parenteral nutrition and related complications. Therefore, the need for better treatments for CD patients remains unmet. Furthermore, the need for novel therapies to treat or improve approximately 180 diseases, including UC and CD, as well as other TL1A-mediated diseases and conditions, has also remained unmet. Given the important functions of TL1A in both innate and adaptive immune-mediated inflammatory responses, therapies targeting TL1A could be used for a variety of inflammation-related and fibrosis-related diseases, bringing significant changes to the treatment of fibrotic and inflammatory diseases. Therefore, the development of TL1A antibodies is necessary. Summary of the Invention

[0005] To address one of the aforementioned technical problems in the prior art, this disclosure provides a TL1A antibody or its antigen-binding fragment. The disclosed antibody or its antigen-binding fragment can bind to TL1A, block the binding of TL1A to DR3, inhibit TL1A-induced TF-1 cell apoptosis, inhibit TL1A-induced NF-κB signaling pathway activation, and inhibit TL1A-induced IFN-γ expression in blood cells. This disclosure also provides the antibody or its antigen-binding fragment for the treatment or prevention of inflammatory diseases or autoimmune diseases (such as colitis) and fibrotic diseases.

[0006] According to one aspect of this disclosure, a TL1A antibody or an antigen-binding fragment thereof is provided, said TL1A antibody or antigen-binding fragment comprising a complementarity-determining region (CDR): a1) having VH-CDR1, VH-CDR2, and VH-CDR3 contained in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO: 2; and / or, having VL-CDR1, VL-CDR2, and VL-CDR3 contained in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO: 4; or a2) VH-CDR1, VH-CDR2, and VH-CDR3 contained in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO: 3; and / or, VL-CDR1, VL-CDR2, and VL-CDR3 contained in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO: 5; or a3) Any amino acid sequence in the CDR sequence of a1) or a2) above may optionally include an amino acid sequence that has been added, deleted, modified and / or substituted with 1-3 amino acid residues and retains TL1A binding activity. The CDR is defined by a definition scheme of Kabat, Chothia, IMGT, Martin, Contact, AbM or a combination thereof.

[0007] In some embodiments, the antibody or its antigen-binding fragment specifically binds to TL1A. In some embodiments, the antibody or its antigen-binding fragment specifically binds to TL1A or fragments thereof derived from humans or cynomolgus monkeys.

[0008] In some embodiments, the antibody or its antigen-binding fragment can block the binding of TL1A to DR3, inhibit TL1A-induced TF-1 cell apoptosis, inhibit TL1A-induced NF-κB signaling pathway activation, and inhibit TL1A-induced expression of IFN-γ in blood cells.

[0009] TL1A antibody or antigen-binding fragment thereof, wherein the TL1A antibody or antigen-binding fragment thereof comprises: b1) Heavy chain variable regions, including VH-CDR1, VH-CDR2, and VH-CDR3; and The light chain variable region includes VL-CDR1, VL-CDR2 and VL-CDR3; The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, and VL-CDR3 are shown in SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10, respectively. The amino acid sequence of VL-CDR2 is YTS, and the CDR is defined according to the IMGT scheme; or The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are shown in SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 10, respectively, and the CDRs are defined according to the Kabat scheme; or The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are shown in SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 10, respectively, and the CDRs are defined according to the Chothia scheme; or The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are shown in SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, and SEQ ID NO: 23, respectively. The CDRs are defined using the Contact definition scheme; or The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are shown in SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 10, respectively, and the CDRs are defined using the AbM definition scheme; or b2) Heavy chain variable regions, including VH-CDR1, VH-CDR2, and VH-CDR3; and The light chain variable region includes VL-CDR1, VL-CDR2 and VL-CDR3; The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, and VL-CDR3 are shown in SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, and SEQ ID NO: 30, respectively. The amino acid sequence of VL-CDR2 is YSS. The CDR is defined according to the IMGT definition scheme; or The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are shown in SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 30, respectively, and the CDRs are defined according to the Kabat scheme; or The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are shown in SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 30, respectively, and the CDRs are defined according to the Chothia scheme; or The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are shown in SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, and SEQ ID NO: 43, respectively. The CDRs are defined using the Contact definition scheme; or The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are shown in SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 30, respectively, and the CDRs are defined using the AbM definition scheme; or b3) Heavy chain variable region; and Light chain variable region, Wherein, compared with any of the heavy chain variable regions and / or light chain variable regions described in b1)-b2), at least one CDR contains a mutation, wherein the mutation is a substitution, deletion or addition of one or more amino acids; preferably, the substitution is a conservative substitution.

[0010] In some embodiments, the heavy chain variable region of the TL1A antibody or its antigen-binding fragment may include the framework region of the heavy chain variable region.

[0011] In some embodiments, the framework region of the heavy chain variable region may include the framework region of the heavy chain variable region or a mutant thereof derived from immunoglobulins of mice, primates, cattle, horses, pigs, sheep, goats, dogs, cats, rabbits, camels, donkeys, deer, mink, chickens, ducks, or geese. In some embodiments, the framework region of the heavy chain variable region may include the framework region of the heavy chain variable region or a mutant thereof derived from immunoglobulins of mice, rats, lemurs, macaques, chimpanzees, or humans, preferably including the framework region of the heavy chain variable region or a mutant thereof derived from human or mouse immunoglobulins, and more preferably including the framework region of the heavy chain variable region or a mutant thereof derived from human immunoglobulins.

[0012] In some embodiments, the light chain variable region of the TL1A antibody or its antigen-binding fragment may include the framework region of the light chain variable region.

[0013] In some embodiments, the framework region of the light chain variable region may include the framework region of the light chain variable region or a mutant thereof derived from immunoglobulins of mice, primates, cattle, horses, pigs, sheep, goats, dogs, cats, rabbits, camels, donkeys, deer, mink, chickens, ducks, or geese. In some embodiments, the framework region of the light chain variable region may include the framework region of the light chain variable region or a mutant thereof derived from immunoglobulins of mice, rats, lemurs, macaques, chimpanzees, or humans, preferably including the framework region of the light chain variable region or a mutant thereof derived from human or mouse immunoglobulins, and more preferably including the framework region of the light chain variable region or a mutant thereof derived from human immunoglobulins.

[0014] In some embodiments, the TL1A antibody or its antigen-binding fragment comprises: The heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 2, 3, 50 or SEQ ID NO: 56, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity with it; and / or, the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 4, 5, 51 or SEQ ID NO: 57, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity with it.

[0015] In some embodiments, the antibody or its antigen-binding fragment comprises: 1) A heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 2, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 4, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; 2) Heavy chain variable region, comprising an amino acid sequence as shown in SEQ ID NO: 3, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, light chain variable region, comprising an amino acid sequence as shown in SEQ ID NO: 5, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; 3) Heavy chain variable region, comprising an amino acid sequence as shown in SEQ ID NO: 50, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, light chain variable region, comprising an amino acid sequence as shown in SEQ ID NO: 51, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; 4) Heavy chain variable region, comprising an amino acid sequence as shown in SEQ ID NO: 56, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, light chain variable region, comprising an amino acid sequence as shown in SEQ ID NO: 57, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0016] In some embodiments, the TL1A antibody or its antigen-binding fragment may further include a heavy chain constant region and / or a light chain constant region.

[0017] In some embodiments, the heavy chain constant region may include the heavy chain constant region or a mutant thereof derived from immunoglobulins of mice, primates, cattle, horses, pigs, sheep, goats, dogs, cats, rabbits, camels, donkeys, deer, mink, chickens, ducks, or geese. In some embodiments, the heavy chain constant region may include the heavy chain constant region or a mutant thereof derived from immunoglobulins of mice, rats, lemurs, macaques, chimpanzees, or humans, preferably including the heavy chain constant region or a mutant thereof derived from human or mouse immunoglobulins, and more preferably including the heavy chain constant region or a mutant thereof derived from human immunoglobulins.

[0018] In some embodiments, the light chain constant region may include a light chain constant region or a mutant thereof derived from immunoglobulins of mice, primates, cattle, horses, pigs, sheep, goats, dogs, cats, rabbits, camels, donkeys, deer, mink, chickens, ducks, or geese. In some embodiments, the light chain constant region may include a light chain constant region or a mutant thereof derived from immunoglobulins of mice, rats, lemurs, macaques, chimpanzees, or humans, preferably a light chain constant region or a mutant thereof derived from human or mouse immunoglobulins, and more preferably a light chain constant region or a mutant thereof derived from human immunoglobulins.

[0019] In some embodiments, the heavy chain constant region includes heavy chain constant regions derived from IgA1, IgA2, IgD, IgE, IgG1, IgG2, IgG3, IgG4, or IgM immunoglobulins, preferably including heavy chain constant regions derived from IgG1 immunoglobulin. In some embodiments, the light chain constant region includes light chain constant regions derived from κ and λ immunoglobulins, preferably including light chain constant regions derived from κ immunoglobulin.

[0020] In some embodiments, the heavy chain constant region includes the human IgG1 heavy chain constant region or a mutant thereof.

[0021] In some embodiments, the mutant of the human IgG1 heavy chain constant region includes: 1) having the following substitutions compared to its derived wild-type sequence: M252Y, S254T and / or T256E (preferably M252Y, S254T and T256E); and / or 2) having the following substitutions compared to its derived wild-type sequence: M428L and / or N434S (preferably M428L and N434S); wherein the amino acid positions mentioned above are positions according to the EU numbering system.

[0022] In some embodiments, the heavy chain constant region includes an amino acid sequence as shown in SEQ ID NO: 52, 54 or 55, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity with it.

[0023] In some embodiments, the light chain constant region comprises an amino acid sequence as shown in SEQ ID NO: 53, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0024] In some embodiments, the TL1A antibody or its antigen-binding fragment may include, but is not limited to, Fab fragments, Fab' fragments, F(ab')2 fragments, Fv fragments, single-chain Fv (scFv), dsFv, and Fd fragments.

[0025] In some embodiments, the TL1A antibody or its antigen-binding fragment includes, but is not limited to, murine antibodies, chimeric antibodies, humanized antibodies, and fully human antibodies.

[0026] In some embodiments, the TL1A antibody or its antigen-binding fragment comprises: a heavy chain and / or a light chain; q1) The heavy chain comprises amino acids 19-472 of SEQ ID NO: 46, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it, and / or the light chain comprises amino acids 19-232 of SEQ ID NO: 47, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; or q2) The heavy chain comprises amino acids 19-473 of SEQ ID NO: 48, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it, and / or the light chain comprises amino acids 19-232 of SEQ ID NO: 49, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; or q3) The heavy chain comprises the amino acid sequence shown in SEQ ID NO: 58, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it, and / or the light chain comprises the amino acid sequence shown in SEQ ID NO: 59, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; or q4) The heavy chain comprises the amino acid sequence shown in SEQ ID NO: 60, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it, and / or the light chain comprises the amino acid sequence shown in SEQ ID NO: 61, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; or q5) The heavy chain comprises the amino acid sequence shown in SEQ ID NO: 62, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it, and / or the light chain comprises the amino acid sequence shown in SEQ ID NO: 63, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; or q6) The heavy chain comprises the amino acid sequence shown in SEQ ID NO:64, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it, and / or the light chain comprises the amino acid sequence shown in SEQ ID NO:65, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; or q7) The heavy chain comprises the amino acid sequence shown in SEQ ID NO: 66, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it, and / or the light chain comprises the amino acid sequence shown in SEQ ID NO: 67, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; or q8) The heavy chain comprises an amino acid sequence as shown in SEQ ID NO: 68, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it, and / or the light chain comprises an amino acid sequence as shown in SEQ ID NO: 69, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0027] According to another aspect of this disclosure, a chimeric antigen receptor is provided, the chimeric antigen receptor comprising an antigen-binding domain, a transmembrane domain and an intracellular signal transduction domain, wherein the antigen-binding domain comprises the TL1A antibody of this disclosure or an antigen-binding fragment thereof.

[0028] According to another aspect of this disclosure, an isolated nucleic acid molecule is provided that encodes the TL1A antibody of this disclosure or its antigen-binding fragment, or the chimeric antigen receptor described above in this disclosure.

[0029] According to another aspect of this disclosure, a carrier is provided that includes the nucleic acid molecules described above.

[0030] In some embodiments, the vector may be an expression vector. In some embodiments, the expression vector may include eukaryotic cell expression vectors and / or prokaryotic cell expression vectors. In some embodiments, the eukaryotic expression vector includes, for example, but not limited to, yeast expression vectors, mammalian expression vectors, and insect expression vectors. For example, the expression vector may include, but is not limited to, plasmids, retroviral vectors, lentiviral vectors, bacteriophage vectors, adenovirus vectors, adeno-associated vectors, or herpes simplex vectors.

[0031] In some embodiments, the carrier may be selected from nanoparticles, liposomes, exogenous bodies, microbubbles, or gene guns.

[0032] According to another aspect of this disclosure, a cell is provided comprising the TL1A antibody or antigen-binding fragment thereof disclosed herein, the chimeric antigen receptor disclosed herein, the nucleic acid molecule disclosed herein, or the vector disclosed herein.

[0033] In some embodiments, the cells do not involve reproductive material.

[0034] In some embodiments, the cell can be a host cell conventionally used in the art, as long as the expression vector stably expresses the carried nucleic acid molecule as the TL1A antibody or its antigen-binding fragment or chimeric antigen receptor disclosed herein. In some embodiments, the host cell can be a prokaryotic cell and / or a eukaryotic cell. The prokaryotic cell may include, for example, *Escherichia coli*, and the eukaryotic cell may include, for example, CHO cells, HEK293 cells, BHK cells, NSO cells, SP2 / 0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells or hybridoma cells, yeast cells, and insect cells.

[0035] In some embodiments, the cells may be immune cells. In some embodiments, the immune cells may include, but are not limited to, monocytes (e.g., T cells) and macrophages. In these embodiments, the immune cells may express the chimeric antigen receptors described above in this disclosure.

[0036] According to another aspect of this disclosure, a pharmaceutical composition is provided comprising the TL1A antibody or antigen-binding fragment thereof disclosed herein, the chimeric antigen receptor disclosed herein, the nucleic acid molecule disclosed herein, the carrier disclosed herein, or the cell disclosed herein. In some embodiments, the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier.

[0037] In some embodiments, the pharmaceutical composition can be used to prevent or treat TL1A-related diseases. In some embodiments, the TL1A-related diseases include fibrotic diseases, as well as inflammatory diseases or autoimmune diseases. In some embodiments, the inflammatory diseases or autoimmune diseases may include, but are not limited to, asthma, arthritis, multiple sclerosis, inflammatory bowel diseases (such as ulcerative colitis and Crohn's disease), chronic obstructive pulmonary disease, glomerulonephritis, psoriasis, allergic dermatitis, bronchitis, allergic rhinitis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, and systemic sclerosis-associated interstitial lung disease. In some embodiments, the fibrotic disease may include, but is not limited to, systemic fibrotic diseases (e.g., systemic sclerosis, multifocal fibrosis, renal systemic fibrosis, scleroderma graft-versus-host disease) and organ-specific fibrotic diseases (e.g., pulmonary fibrosis (e.g., idiopathic pulmonary fibrosis, silica-induced pneumoconiosis, asbestos-induced pulmonary fibrosis, chemotherapy-induced pulmonary fibrosis), cardiac fibrosis (e.g., hypertension-associated cardiac fibrosis, post-myocardial infarction, Chagas disease-induced myocardial fibrosis), and renal fibrosis (e.g., diabetes mellitus). Diseases and hypertensive nephropathy, urinary tract obstruction-induced renal fibrosis, inflammatory / autoimmune-induced renal fibrosis, aristolochic acid nephropathy, polycystic kidney disease), hepatic and portal vein fibrosis (e.g., alcoholic and non-alcoholic liver fibrosis, hepatitis C-induced liver fibrosis, primary biliary cirrhosis, parasite-induced liver fibrosis), radiation-induced fibrosis, bladder fibrosis, intestinal fibrosis, peritoneal sclerosis, diffuse fasciitis (e.g., localized scleroderma, keloids, Dupuytren's disease, Peroni's disease, myelofibrosis, oral submucosal fibrosis), scarring.

[0038] According to another aspect of this disclosure, a product is provided comprising the TL1A antibody or antigen-binding fragment thereof described above, wherein the product comprises at least one of reagents, chips, test strips, and detection kits. In some embodiments, the product can be used for: 1) detecting the presence or amount of TL1A in a sample; 2) diagnosing or prognostically assessing TL1A-related diseases.

[0039] In some embodiments, the TL1A-related diseases include fibrotic diseases, as well as inflammatory diseases or autoimmune diseases. In some embodiments, the inflammatory diseases or autoimmune diseases may include, but are not limited to, asthma, arthritis, multiple sclerosis, inflammatory bowel diseases (such as ulcerative colitis and Crohn's disease), chronic obstructive pulmonary disease, glomerulonephritis, psoriasis, allergic dermatitis, bronchitis, allergic rhinitis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, and systemic sclerosis-associated interstitial lung disease. In some embodiments, the fibrotic disease may include, but is not limited to, systemic fibrotic diseases (e.g., systemic sclerosis, multifocal fibrosis, renal systemic fibrosis, scleroderma graft-versus-host disease) and organ-specific fibrotic diseases (e.g., pulmonary fibrosis (e.g., idiopathic pulmonary fibrosis, silica-induced pneumoconiosis, asbestos-induced pulmonary fibrosis, chemotherapy-induced pulmonary fibrosis), cardiac fibrosis (e.g., hypertension-associated cardiac fibrosis, post-myocardial infarction, Chagas disease-induced myocardial fibrosis), and renal fibrosis (e.g., diabetes mellitus). Diseases and hypertensive nephropathy, urinary tract obstruction-induced renal fibrosis, inflammatory / autoimmune-induced renal fibrosis, aristolochic acid nephropathy, polycystic kidney disease), hepatic and portal vein fibrosis (e.g., alcoholic and non-alcoholic liver fibrosis, hepatitis C-induced liver fibrosis, primary biliary cirrhosis, parasite-induced liver fibrosis), radiation-induced fibrosis, bladder fibrosis, intestinal fibrosis, peritoneal sclerosis, diffuse fasciitis (e.g., localized scleroderma, keloids, Dupuytren's disease, Peroni's disease, myelofibrosis, oral submucosal fibrosis), scarring.

[0040] In some embodiments, the TL1A antibody or its antigen-binding fragment disclosed herein may be conjugated to a detectable marker.

[0041] In some embodiments, the detectable marker may be a fluorescent or luminescent marker. In some embodiments, the detectable marker may be selected from, for example, any one of acridine ester, acridine sulfonamide, luminol, isoluminol, horseradish peroxidase, and alkaline phosphatase.

[0042] According to another aspect of this disclosure, a method for detecting the presence or content of TL1A in a sample is provided, wherein the sample is contacted with the TL1A antibody or antigen-binding fragment of this disclosure under conditions that allow the TL1A antibody or antigen-binding fragment thereof described in this disclosure to form a complex with TL1A, and the formation of the complex is detected, the method being for non-diagnostic or therapeutic purposes.

[0043] According to another aspect of this disclosure, the use of the above-described TL1A antibody or antigen-binding fragment thereof, the above-described chimeric antigen receptor, the above-described nucleic acid molecule, the above-described carrier, the above-described cell, or the above-described pharmaceutical composition in the preparation of a medicament for the prevention or treatment of TL1A-related diseases is provided.

[0044] According to another aspect of this disclosure, a method for preventing or treating TL1A-related diseases is provided, the method comprising administering to a subject in need a therapeutically effective amount of the TL1A antibody or antigen-binding fragment thereof of this disclosure, the chimeric antigen receptor, the nucleic acid molecule, the carrier, the cell, or the pharmaceutical composition thereof.

[0045] In some embodiments, the TL1A-related diseases include fibrotic diseases, as well as inflammatory diseases or autoimmune diseases. In some embodiments, the inflammatory diseases or autoimmune diseases may include, but are not limited to, asthma, arthritis, multiple sclerosis, inflammatory bowel diseases (such as ulcerative colitis and Crohn's disease), chronic obstructive pulmonary disease, glomerulonephritis, psoriasis, allergic dermatitis, bronchitis, allergic rhinitis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, and systemic sclerosis-associated interstitial lung disease. In some embodiments, the fibrotic disease may include, but is not limited to, systemic fibrotic diseases (e.g., systemic sclerosis, multifocal fibrosis, renal systemic fibrosis, scleroderma graft-versus-host disease) and organ-specific fibrotic diseases (e.g., pulmonary fibrosis (e.g., idiopathic pulmonary fibrosis, silica-induced pneumoconiosis, asbestos-induced pulmonary fibrosis, chemotherapy-induced pulmonary fibrosis), cardiac fibrosis (e.g., hypertension-associated cardiac fibrosis, post-myocardial infarction, Chagas disease-induced myocardial fibrosis), and renal fibrosis (e.g., diabetes mellitus). Diseases and hypertensive nephropathy, urinary tract obstruction-induced renal fibrosis, inflammatory / autoimmune-induced renal fibrosis, aristolochic acid nephropathy, polycystic kidney disease), hepatic and portal vein fibrosis (e.g., alcoholic and non-alcoholic liver fibrosis, hepatitis C-induced liver fibrosis, primary biliary cirrhosis, parasite-induced liver fibrosis), radiation-induced fibrosis, bladder fibrosis, intestinal fibrosis, peritoneal sclerosis, diffuse fasciitis (e.g., localized scleroderma, keloids, Dupuytren's disease, Peroni's disease, myelofibrosis, oral submucosal fibrosis), scarring. Attached Figure Description

[0046] Figure 1 The binding activity of Y10-L5 to human TL1A protein was demonstrated.

[0047] Figure 2 The binding activity of Y10-L15 to human TL1A protein was demonstrated.

[0048] Figure 3 The inhibitory activity of Y10-L5 on the binding of human TL1A to human DR3 protein was demonstrated.

[0049] Figure 4 The inhibitory activity of Y10-L5 on the binding of human TL1A to CHO-DR3 was demonstrated.

[0050] Figure 5 The inhibitory activity of Y10-L15 on the binding of human TL1A to CHO-DR3 was demonstrated.

[0051] Figure 6 The inhibitory activity of Y10-L5 on TL1A-induced apoptosis in TF-1 cells was demonstrated.

[0052] Figure 7 The inhibitory activity of Y10-L5 on TL1A-induced NF-κB signaling pathway activation was demonstrated.

[0053] Figure 8 The inhibitory activity of Y10-L15 on TL1A-induced NF-κB signaling pathway activation was demonstrated.

[0054] Figure 9 The binding activity of humanized antibodies Y10-L5VH1-1VL1-1, Y10-L5VH1-1VL1-1 (YTE), and Y10-L5VH1-1VL1-1 (LS) to human TL1A protein was demonstrated.

[0055] Figure 10 The binding activity of humanized antibodies Y10-L15VH1VL1, Y10-L15VH1VL1 (YTE), and Y10-L15VH1VL1 (LS) to human TL1A protein was demonstrated.

[0056] Figure 11 The binding activity of humanized antibodies Y10-L5VH1-1VL1-1, Y10-L5VH1-1VL1-1 (YTE), and Y10-L5VH1-1VL1-1 (LS) to the FcRn receptor was demonstrated.

[0057] Figure 12 The binding activity of humanized antibodies Y10-L15VH1VL1, Y10-L15VH1VL1 (YTE), and Y10-L15VH1VL1 (LS) to the FcRn receptor was demonstrated.

[0058] Figure 13 The inhibitory activity of humanized antibodies Y10-L5VH1-1VL1-1, Y10-L5VH1-1VL1-1 (YTE), and Y10-L5VH1-1VL1-1 (LS) on the binding of human TL1A to CHO-DR3 was demonstrated.

[0059] Figure 14 The inhibitory activity of humanized antibodies Y10-L15VH1VL1, Y10-L15VH1VL1 (YTE), and Y10-L15VH1VL1 (LS) on the binding of human TL1A to CHO-DR3 was demonstrated.

[0060] Figure 15 The thermal stability of humanized antibodies Y10-L5VH1-1VL1-1 and Y10-L15VH1VL1 was demonstrated.

[0061] Figure 16 The thermal stability of the humanized antibodies Y10-L15VH1VL1 (YTE), Y10-L15VH1VL1 (LS), Y10-L5VH1-1VL1-1 (YTE), and Y10-L5VH1-1VL1-1 (LS) was demonstrated.

[0062] Figure 17 The inhibitory activity of the humanized antibody Y10-L5VH1-1VL1-1 on TL1A-induced apoptosis in TF-1 cells was demonstrated.

[0063] Figure 18 The inhibitory activity of the humanized antibody Y10-L15VH1VL1 on TL1A-induced apoptosis in TF-1 cells was demonstrated.

[0064] Figure 19 The inhibitory activity of humanized antibodies Y10-L15VH1VL1 (YTE), Y10-L15VH1VL1 (LS), Y10-L5VH1-1VL1-1 (YTE), and Y10-L5VH1-1VL1-1 (LS) on TL1A-induced apoptosis in TF-1 cells was demonstrated.

[0065] Figure 20 The inhibitory activity of humanized antibodies Y10-L5VH1-1VL1-1, Y10-L5VH1-1VL1-1 (YTE), and Y10-L5VH1-1VL1-1 (LS) on TL1A-induced NF-κB signaling pathway activation was demonstrated.

[0066] Figure 21 The inhibitory activity of humanized antibodies Y10-L15VH1VL1, Y10-L15VH1VL1 (YTE), and Y10-L15VH1VL1 (LS) on TL1A-induced NF-κB signaling pathway activation was demonstrated.

[0067] Figure 22 The inhibitory activity of the humanized antibody against IFN-γ expression induced by recombinant TL1A in human PBMCs was demonstrated.

[0068] Figure 23 The inhibitory activity of the humanized antibody Y10-L15VH1VL1(LS) on the expression of IFN-γ in whole blood cells stimulated by immune complexes was demonstrated.

[0069] Figure 24 The inhibitory activity of humanized antibodies Y10-L5VH1-1VL1-1 (YTE) and Y10-L5VH1-1VL1-1 (LS) on the expression of IFN-γ in whole blood cells stimulated by immune complexes was demonstrated.

[0070] Figure 25 The binding activity of the humanized antibody to mouse TL1A was demonstrated.

[0071] Figure 26 The binding activity of the humanized antibody to cynomolgus monkey TL1A was demonstrated.

[0072] Figure 27 The binding activity of the humanized antibody to rat TL1A was demonstrated.

[0073] Figure 28 The study showed that humanized antibodies can increase the length of the colon and rectum in mice with acute enteritis.

[0074] Figure 29 The study showed that humanized antibodies can reduce intestinal mucosal structural damage in mice with acute enteritis.

[0075] Figure 30 The drug concentrations in mouse serum at different time points are shown.

[0076] Figure 31 The study showed that humanized antibodies can increase the body weight of mice with acute enteritis.

[0077] Figure 32 The study showed that humanized antibodies can reduce the disease activity index in mice with acute enteritis.

[0078] Figure 33 The study showed that humanized antibodies can reduce the expression of inflammatory factors in the colon tissue of mice with acute enteritis. Detailed Implementation

[0079] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. The specific embodiments described herein are for illustrative purposes only and are not intended to limit the invention in any way. Furthermore, descriptions of well-known structures and techniques are omitted in the following description to avoid unnecessarily obscuring the concepts of this disclosure. Such structures and techniques have also been described in many publications.

[0080] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly used in the field to which this invention pertains. For the purposes of interpreting this specification, the following definitions will apply, and where appropriate, terms used in the singular will also include the plural forms, and vice versa.

[0081] Unless the context clearly indicates otherwise, the terms “a” and “an” as used herein include plural references. For example, reference to “a cell” includes multiple such cells and equivalents known to those skilled in the art, etc.

[0082] As used herein, the term "about" indicates a range of ±20% of the following value. In some embodiments, the term "about" indicates a range of ±10% of the following value. In some embodiments, the term "about" indicates a range of ±5% of the following value.

[0083] As used in this article, "antibody" refers to a globulin produced by plasma cells, which are formed from the proliferation and differentiation of B lymphocytes in response to antigen stimulation. Antibodies specifically bind to the corresponding antigens and mediate immune effects. They are mainly found in serum and body fluids and are important immune molecules mediating humoral immunity. Antibodies can encompass various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, single-chain molecules, and antigen-binding fragments. The chemical basis of antibodies is immunoglobulin (Ig).

[0084] As used herein, the term "monoclonal antibody" refers to antibodies derived from a substantially homogeneous group of antibodies, meaning that, apart from possible trace amounts of variant antibodies (e.g., containing naturally occurring mutations or generated during the production of the monoclonal antibody formulation, typically present in small quantities), the individual antibodies within the group are identical and / or bind to the same epitopes. Unlike polyclonal antibody formulations, which typically comprise different antibodies targeting different antigenic determinants (epitaxes), each monoclonal antibody in a monoclonal antibody formulation targets a single determinant on the antigen.

[0085] The terms “full-length antibody” and “intact antibody” as used herein are used interchangeably to refer to antibodies that are structurally similar to natural antibodies. “Natural antibody” refers to a naturally occurring immunoglobulin molecule. For example, natural IgG antibodies are heterotetrameric glycoproteins of approximately 150,000 Daltons, composed of two light chains and two heavy chains linked by disulfide bonds. From the N-terminus to the C-terminus, each heavy chain has a variable region (VH) (also called a variable heavy chain domain or heavy chain variable domain) and three constant domains (CH1, CH2, and CH3) (also called heavy chain constant regions). From the N-terminus to the C-terminus, each light chain has a variable region (VL) (also called a variable light chain domain or light chain variable domain) and a light chain constant domain (CL) (also called light chain constant regions). The heavy chain of an antibody can be one of five types: α (IgA), δ (IgD), ε (IgE), γ (IgG), or μ (IgM), and can be further subdivided into subtypes such as γ1 (IgG1), γ2 (IgG2), γ3 (IgG3), γ4 (IgG4), α1 (IgA1), and α2 (IgA2). The light chain of an antibody, based on the amino acid sequence of its constant domain, can be one of two types: k-light chains and λ-light chains.

[0086] Within the light and heavy chains, variable and constant regions are linked by a "J" region containing approximately 12 or more amino acid residues, and the heavy chain also contains a "D" region containing approximately 3 or more amino acids. Each heavy chain consists of a heavy chain variable region (VH) and a heavy chain constant region (CH). The heavy chain constant region consists of three domains (CH1, CH2, and CH3). Each light chain consists of a light chain variable region (VL) and a light chain constant region (CL). The light chain constant region consists of one domain, CL. The constant region of an antibody mediates the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system.

[0087] As used herein, the term "variable region" or "variable domain" refers to the domain of the antibody heavy or light chain involved in the binding of the antigen-binding molecule to the antigen. The variable domains (VH and VL, respectively) of the heavy and light chains of natural antibodies typically have similar structures, with each domain containing four conserved framework regions (FR1-4) and three hypervariable regions (HVR1-3), arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. A single VH or VL domain is sufficient to confer antigen-binding specificity. The three HVRs within the VH and VL domains together constitute the antigen-binding site of Ig, which can bind complementary to the corresponding antigenic epitope; therefore, the HVRs are also called complementarity-determining regions (CDRs), denoted as CDR1, CDR2, and CDR3, respectively. The VH or VL chain of the antibody may further contain all or part of the constant regions of the heavy or light chain.

[0088] The CDR of the antibody or antigen-binding fragment thereof disclosed herein can be determined according to various numbering systems known in the art. In some embodiments, the CDR contained in the antibody or antigen-binding fragment thereof disclosed herein is preferably determined by the IMGT, Kabat, Chothia, Contact, or AbM numbering system.

[0089] The following general rules (published at www.bioinf.org.uk: Professor Andrew C. Martin's research group) can be used to define CDRs in antibody sequences, which include amino acids that specifically interact with the amino acids that form the antigenic epitope that the antibody binds to. In rare cases, these generally constant features may not appear; however, Cys residues are the most conserved feature.

[0090] As used herein, the term "variable" refers to the fact that certain segments of the variable domain are generally different in sequence between antibodies. The V domain mediates antigen binding and defines the specificity of a particular antibody for its specific antigen. However, variability is not uniformly distributed throughout the variable domain, but is concentrated in three segments called hypervariable regions (HVRs) within the variable domains of the light and heavy chains. The relatively highly conserved portions of the variable domain are called frame regions (FRs). The variable domains of the native heavy and light chains each contain four FRs, mostly in a β-sheet configuration, linked by three HVRs that form loops and, in some cases, form part of a β-sheet structure. The HVRs in each chain are tightly held together by the FRs and, together with the HVRs of other chains, contribute to the formation of the antibody's antigen-binding site. The constant domain does not directly participate in antibody-antigen binding but has other effector functions, such as participating in antibody-dependent cytotoxicity.

[0091] As used herein, the term "framework" or "FR" refers to the variable domain residues other than the hypervariable region (HVR) residues. A variable domain FR typically consists of four FR domains: FR1, FR2, FR3, and FR4. Therefore, the HVR and FR sequences typically appear in the VH (or VL) in the following sequence: VH(VL)-FR1-VH(VL)-CDR1-VH(VL)-FR2-VH(VL)-CDR2-VH(VL)-FR3-VH(VL)-CDR3-VH(VL)-FR4.

[0092] Antibody "classes" refer to the types of constant structural domains or constant regions possessed by the antibody's heavy chain. Based on differences in heavy chain structure and antigenicity, they can be classified into five classes: μ chain, γ chain, α chain, δ chain, and ε chain. Immunoglobulins composed of different heavy and light chains are respectively called IgA, IgD, IgE, IgG, and IgM. Even within the same class of Ig, the amino acid composition of the hinge region and the number and position of disulfide bonds in the heavy chain differ, thus further subdividing the same class of Ig into different subclasses. For example, human IgG can be divided into IgG1–IgG4; IgA can be divided into IgA1 and IgA2. Based on differences in light chain structure and antigenicity, immunoglobulin (Ig) light chains are divided into κ (kappa) chains and λ (lambda) chains, thus classifying Ig into two types: κ type and λ type.

[0093] "Humanized antibodies" comprise amino acid residues from non-human HVRs and amino acid residues from human FRs. In some embodiments, humanized antibodies comprise at least one, typically two, variable domains, wherein all or substantially all HVRs (e.g., CDRs) correspond to the HVRs of the non-human antibody, and all or substantially all FRs correspond to the FRs of the human antibody. Humanized antibodies may optionally comprise at least a portion of the antibody constant region derived from a human antibody. Antibodies in a "humanized form," such as non-human antibodies, refer to antibodies that have undergone humanization.

[0094] "Humanized antibodies" have an amino acid sequence that corresponds to that of antibodies produced by humans or human cells, or derived from non-human antibodies using sequences encoded by human antibody libraries or other human antibodies. This definition of human antibodies specifically excludes humanized antibodies containing non-human antigen-binding residues.

[0095] As used herein, the term "Fc domain" or "Fc region" is used to define a C-terminal region of an antibody heavy chain containing at least a portion of a constant region. This term includes native sequence Fc regions and variant Fc regions. An IgG Fc region contains an IgG CH2 domain and an IgG CH3 domain. The CH2 domain as used herein may be a native sequence CH2 domain or a variant CH2 domain. The CH3 region as used herein may be a native sequence CH3 domain or a variant CH3 domain. The CH2 domain may contain one or more mutations that reduce or eliminate the binding of the CH2 domain to one or more Fcγ receptors (e.g., FcγRI, FcγRIIa, FcγRIIb, FcγRIII) and / or complement. It is presumed that reducing or eliminating binding to Fc receptor γ will reduce or eliminate antibody-mediated ADCC. Similarly, reducing or eliminating binding to complement is expected to reduce or eliminate antibody-mediated CDC. Mutations that reduce or eliminate the binding of the CH2 domain to one or more Fcγ receptors and / or complement are known in the art. These mutations include the so-called "LALA mutation," which involves replacing leucine residues at positions 1.3 and 1.2 of the IMGT site in the CH2 domain with alanine (L1.3A and L1.2A). Alternatively, it is also known to generate α-glycosylated antibodies by mutating asparagine (N) at position 84.4 of the IMGT site in the CH2 domain to alanine, glycine, or glutamine (N84.4A, N84.4G, or N84.4Q), thereby mutating conserved N-chain glycosylation sites to reduce IgG1 effector function. As another alternative, complement activation (C1q binding) and ADCC are known to be reduced by mutating proline at position 114 of the IMGT site in the CH2 domain to alanine or glycine (P114A or P114G). These mutations can be combined to produce antibody molecules with further reduced or no ADCC or CDC activity.

[0096] "Regions equivalent to the Fc region of immunoglobulins" include variants of the naturally occurring alleles of the Fc region of immunoglobulins, as well as modified variants that have the ability to produce substitutions, additions, or deletions that substantially do not diminish the function of immunoglobulins-mediated effectors, such as antibody-dependent cytotoxicity. For example, one or more amino acids can be deleted from the N-terminus or C-terminus of the Fc region of an immunoglobulin without substantially losing its biological function. Such variants can be selected according to general rules known in the art to minimize the impact on activity (see, for example, Bowie, JU et al., Science 247:1306-10 (1990)).

[0097] As used in this article, the term "effector function" refers to the biological activity attributable to the Fc region of an antibody, which varies with antibody isotype. Examples of antibody effector functions include: C1q binding and complement-dependent cytotoxicity (CDC), Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent phagocytosis (ADCP), cytokine secretion, antigen uptake by immune complex-mediated antigen-presenting cells, downregulation of cell surface receptors (e.g., B cell receptors), and B cell activation.

[0098] As used herein, the terms “polynucleotide,” “nucleic acid,” or “nucleotide sequence” refer to isolated nucleic acid molecules or constructs, such as messenger RNA (mRNA), virus-derived RNA, or plasmid DNA (pDNA). Polynucleotides may contain conventional phosphodiester bonds or unconventional bonds (such as amide bonds, as found in peptide nucleic acids (PNAs)). The term “nucleic acid molecule” refers to any one or more nucleic acid segments, such as DNA or RNA fragments, present in a polynucleotide.

[0099] An "antibody fragment" or "antigen-binding fragment" contains a portion of a complete antibody that retains the antibody's antigen-binding activity. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv; bisomatic antibodies, trisomatic antibodies, tetrasomatic antibodies, cross-Fab fragments; linear antibodies; single-chain antibody molecules (e.g., scFv); and multispecific antibodies formed from antibody fragments and single-domain antibodies (single-domain antibodies).

[0100] As used herein, the terms "antigen-binding domain" or "antigen-binding site" refer to the portion of an antigen-binding molecule that specifically binds to an antigenic determinant. More specifically, the term "antigen-binding domain" refers to a portion of an antibody containing a region that specifically binds to and is complementary to a portion or all of the antigen. In cases where the antigen molecule is large, the antigen-binding molecule may bind only a specific portion of the antigen, called an epitope. The antigen-binding domain may be provided by, for example, one or more variable domains (also called variable regions). Preferably, the antigen-binding domain comprises a variable region (VL) of the antibody light chain and a variable region (VH) of the antibody heavy chain. In one aspect, the antigen-binding domain is capable of binding its antigen and blocking or partially blocking the function of said antigen.

[0101] As used herein, the term "antigenic determinant" is synonymous with "antigen" and "epitope" and refers to a site on a polypeptide macromolecule (e.g., a continuous amino acid sequence or a conformation composed of different regions of non-continuous amino acids) to which an antigen-binding moiety binds, thereby forming an antigen-binding moiety-antigen complex. Antigenic determinants can be present, for example, on the surface of tumor cells, on the surface of microbially infected cells, on the surface of other diseased cells, on the surface of immune cells, in serum, and / or in the extracellular matrix (ECM). Unless otherwise stated, proteins used as antigens in this invention can be any naturally occurring form of protein from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). Antigens can also be human proteins, or antigens can be "full-length," unprocessed proteins, and any form of protein produced by intracellular processing, or naturally occurring protein variants, such as splice variants or allelic variants.

[0102] The specific “binding strength” or “affinity” of an antibody or its antigen-binding fragment to an antigen refers to the strength of the non-covalent interaction between a single binding site and its binding ligand (e.g., antigen), and can be distinguished from unwanted or non-specific binding. The ability of an antigen-binding molecule to bind to a specific antigen can be measured by enzyme-linked immunosorbent assay (ELISA) or other techniques familiar to those skilled in the art, such as surface plasmon resonance (SPR) techniques and conventional binding assays. In one embodiment, for example, as measured by SPR, the degree of binding of the antigen-binding molecule to an unrelated protein is less than about 10% of the degree of binding of the antigen-binding molecule to the antigen. Binding affinity is typically expressed as a dissociation constant (KD), which is the ratio of the dissociation rate constant to the association rate constant (Kd and Ka, respectively). In some embodiments, the dissociation constant (Kd) of the molecule binding to the antigen is ≤100 nM, ≤10 nM, ≤1 nM, ≤0.1 nM, ≤0.01 nM, or ≤0.001 nM (e.g., 10). -7 M or lower, such as 10 -7 M to 10 -13 M, for example, 10 -9 M to 10 -13 M).

[0103] As used herein, the term "isolated" nucleic acid molecule or polynucleotide refers to a nucleic acid molecule, DNA, or RNA, that has been separated from its natural environment. In this invention, the recombinant polynucleotide encoding a polypeptide contained in the vector is also isolated. Other examples of isolated polynucleotides include recombinant polynucleotides in heterologous host cells or polynucleotides purified in solution. Isolated polynucleotides include polynucleotide molecules typically found in cells containing the polynucleotide molecule, but which are located extrachromosomally or at chromosomal locations different from their natural chromosomal locations. Isolated RNA molecules include in vivo or in vitro RNA transcripts of this invention, in positive and negative strand forms, and in double strand forms. The isolated polynucleotides or nucleic acids of this disclosure further include synthetically generated molecules of this type. Additionally, the polynucleotide or nucleic acid may be or may include regulatory elements such as promoters, ribosome binding sites, or transcription terminators.

[0104] As used herein, the terms "vector" or "expression vector" and "expression construct" are used interchangeably. A vector is a DNA molecule to which a specific gene, operably linked, is introduced into a target cell and directed for expression. The vector includes a vector as a self-replicating nucleic acid structure and a vector incorporated into the genome of the host cell into which it has been introduced. The expression vector of the present invention comprises an expression cassette. The expression vector can perform transcription of a large amount of stable mRNA. Once the expression vector is in the target cell, the cellular transcription and / or translation mechanisms generate a ribonucleic acid molecule or protein encoded by the gene.

[0105] As used herein, the term "chimeric antigen receptor" or "CAR" refers to a receptor having desired antigen specificity and signal transduction domains to propagate intracellular signals upon antigen binding. For example, T lymphocytes recognize specific antigens via the interaction of T cell receptors (TCRs) with short peptides presented by class I or II major histocompatibility complex (MHC) molecules. For initial activation and clonal expansion, naïve T cells depend on antigen-presenting cells (APCs) that provide additional co-stimulatory signals. In some embodiments, monocytes and macrophages may be engineered to express, for example, chimeric antigen receptors (CARs). CARs may include antigen-binding domains, transmembrane domains, and intracellular domains. The antigen-binding domain binds to the antigen on the target cell. Examples of cell surface markers of antigens that can be used as antigens binding to the antigen-binding domain of a CAR include those associated with viral, bacterial, parasitic infections, autoimmune diseases, and cancer cells.

[0106] An "effective amount" of a drug is the amount necessary to produce physiological changes in the cells or tissues to which it is administered. An "effective amount" includes the amount sufficient to improve or prevent the symptoms or signs of a medically diagnosed disease. An effective amount also means the amount sufficient to allow or facilitate diagnosis. The effective amount for a particular patient or veterinary subject can vary depending on factors such as the condition to be treated, the patient's overall health, the route and dosage of administration, and the severity of any side effects. An effective amount can be the maximum dose or administration regimen that avoids significant side effects or toxicity.

[0107] The "therapeutic effective amount" of a drug (such as a pharmaceutical composition) refers to the amount necessary to effectively achieve the desired therapeutic or preventive effect in terms of dosage, dosing intervals, and time. For example, a therapeutically effective amount of a drug eliminates, mitigates / reduces, delays, minimizes, or prevents the adverse effects of a disease.

[0108] As used herein, the term “individual” or “subject” refers to a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cattle, sheep, cats, dogs, and horses), primates (e.g., human and non-human primates, such as monkeys), rabbits, and rodents (e.g., mice and rats). Specifically, an individual or subject is a human being.

[0109] As used herein, the term "pharmaceutical composition" refers to a mixture containing one or more antibodies or antigen-binding fragments thereof, chimeric antigen receptors, nucleic acid molecules, carriers, or cells, and other chemical components, such as physiological / pharmaceutical-grade carriers or excipients. The purpose of a pharmaceutical composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredient and the exertment of its biological activity.

[0110] As used herein, the term "pharmaceuticalally acceptable excipient" refers to a component in a pharmaceutical composition that, apart from the active ingredient, is non-toxic to the subject. Pharmaceutically acceptable excipients include, but are not limited to, buffers, stabilizers, and / or preservatives.

[0111] As used herein, the term "treatment" refers to the administration of an oral or topical therapeutic agent, such as a composition comprising any antibody of this disclosure or an antigen-binding fragment thereof, or a nucleic acid molecule encoding an antibody or an antigen-binding fragment thereof, to a patient having one or more TL1A-related diseases or symptoms, the therapeutic agent having a therapeutic effect on these diseases or symptoms. Typically, the therapeutic agent is administered in a treated patient or population in an amount that effectively relieves one or more diseases or symptoms, inducing regression of such symptoms or inhibiting their progression to any clinically measurable extent.

[0112] As used herein, the term “prevention” refers to delaying, suppressing, or preventing the onset of TL1A-related diseases in mammals in which the initiation of fibrotic, inflammatory, or autoimmune diseases has not been confirmed, but susceptibility to such diseases has been identified, for example, through genetic screening or other methods.

[0113] As used herein, the term "detectable marker" encompasses a marker that can be detected directly or indirectly, either attached to the antibody or the bispecific binding protein, or present independently in the kit. Suitable markers include, but are not limited to, molecules detectable by spectroscopic, photochemical, biochemical, immunochemical, chemical, or other physical means. Suitable markers include, but are not limited to, fluorescent dyes (e.g., GFT and its variants, FITC, TRITC, fluorescein, and rhodamine, etc.), electron-dense reagents (e.g., gold), enzymes (e.g., horseradish peroxidase (HRP), alkaline phosphatase (AP), glucose oxidase, β-D-galactosidase, urease, catalase, or glucosyl amylase), molecules containing radionuclides (i.e., radioisotopes), chemiluminescent molecules, electrochemiluminescent molecules, biotin, digoxin / digoxigenin, or haptens, and other entities that are or can be detectable. Antibodies or their antigen-binding fragments or bispecific binding proteins in this disclosure are attached to a "detectable marker," thus being detectably labeled.

[0114] As used herein, the term "conservative substitution" refers to an amino acid substitution that does not adversely affect or alter the intended properties of a protein / peptide containing an amino acid sequence. For example, conservative substitutions can be introduced using standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions include substitutions of amino acid residues with amino acid residues having similar side chains, such as substitutions with residues that are physically or functionally similar to the corresponding amino acid residues (e.g., having similar size, shape, charge, chemical properties, including the ability to form covalent or hydrogen bonds). Families of amino acid residues with similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, and histidine), acidic side chains (e.g., aspartic acid and glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, and tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, and methionine), β-branched side chains (e.g., threonine, valine, and isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, and histidine). Therefore, it is preferable to replace the corresponding amino acid residue with another amino acid residue from the same side chain family. Methods for identifying conserved amino acid substitutions are well known in the art (see, for example, Brummell et al., Biochem. 32:1180-1187 (1993); Kobayashi et al., Protein Eng. 12(10):879-884 (1999); and Burks et al., Proc. Natl Acad. Set USA 94:412-417 (1997), which are incorporated herein by reference).

[0115] The "sequence identity percentage" or "identity percentage" between two polynucleotide or polypeptide sequences refers to the number of identical matching positions shared by sequences within a comparison window, taking into account additions or deletions (i.e., vacancies) that must be introduced for optimal alignment of the two sequences. A matching position is any location where the same nucleotide or amino acid is present in both the target and reference sequences. Vacancies are not nucleotides or amino acids and are not counted in the target sequence. Similarly, vacancies in the reference sequence are not counted because nucleotides or amino acids from the target sequence are counted, but those from the reference sequence are not.

[0116] The percentage of sequence identity can be calculated as follows: determine the number of positions in both sequences where the same amino acid residue or nucleic acid base appears (the number of matching positions), divide the number of matching positions by the total number of positions in the comparison window, and multiply the result by 100 to obtain the percentage of sequence identity. Sequence comparison and determination of the percentage of sequence identity between two sequences can be accomplished using software that is readily available online and downloadable. Suitable software programs are available from various sources for protein and nucleotide sequence alignment. A suitable program for determining the percentage of sequence identity is bl2seq, which is part of the BLAST program suite available from the National Center for Biotechnology Information (NCBI) website (blast.ncbi.nlm.nih.gov). Bl2seq uses either the BLASTN or BLASTP algorithm for comparing two sequences. BLASTN is used to compare nucleic acid sequences, while BLASTP is used to compare amino acid sequences. Other suitable programs are, for example, Needle, Stretcher, Water, or Matcher, which are part of the EMBOSS suite of bioinformatics programs and are also available from the European Institute of Bioinformatics (EBI) at www.ebi.ac.uk / Tools / psa.

[0117] The following embodiments and accompanying drawings are provided to aid in understanding the present invention. However, it should be understood that these embodiments and drawings are for illustrative purposes only and do not constitute any limitation. The actual scope of protection of the present invention is set forth in the claims. It should be understood that any modifications and changes can be made without departing from the spirit of the invention. The reagents and / or kits used in the following embodiments are commercially available or can be synthesized by known methods.

[0118] It should be noted that, unless specific conditions are specified in the examples, experimental conditions should be performed according to standard conditions, manufacturer recommendations, or publicly reported experimental conditions. Reagents or instruments whose manufacturers are not specified are all commercially available, standard products. For reagents whose manufacturers are specified, similar products from other manufacturers are substitutes.

[0119] In the quantitative experiments described below, three replicate experiments were conducted, and the average value of the results was taken.

[0120] Example Example 1: Obtaining anti-human TL1A antibody Recombinant human TL1A (TL1A-ECD, SEQ ID NO:1) (unless otherwise specified, all recombinant proteins are expressed by Taizhou Baiying Biotechnology) and CHO-TL1A (CHO-K1 cells stably overexpressing human TL1A constructed by the inventors) were used as antigens to alternately immunize female Balb / c mice aged 6-8 weeks. The CHO-TL1A construction method is as follows: using the Lipo 2000 (ThermoFisher, 11668019) liposome transfection method, the plasmid carrying the TL1A (uniprot accession number: O95150) target gene was transfected into CHO-K1 cells, and CHO-TL1A cells stably overexpressing TL1A were finally obtained through screening by resistance genes and target genes.

[0121] Spleen cells from immunized mice were fused with SP2 / 0-AG14 cells to form hybridoma cells, and an appropriate amount of the fused cells were seeded into 96-well plates. On days 7-10 post-fusion, the supernatant from each well was collected, and the binding activity of mouse antibodies secreted by the hybridoma cells to human TL1A and their inhibitory activity against human TL1A / DR3 (uniprot accession number: Q93038) were detected by ELISA. Several hybridoma cell lines were obtained that secreted anti-human TL1A antibodies and exhibited good inhibitory activity against TL1A / DR3 binding: 6A7E2 (inhibition rate 86.8%) and 1H119 (inhibition rate 93.0%). Antibody subtype identification results showed that both 6A7E2 and 1H119 were mouse IgG1 antibodies, with the light chain subtype being kappa. Sequencing revealed the heavy chain variable region cDNA sequences and light chain variable region cDNA sequences corresponding to the secreted antibodies (6A7E2, 1H119). The heavy chain amino acid sequences encoded by the heavy chain variable region cDNA sequences of 6A7E2 and 1H119 are shown in SEQ ID NO: 2 and SEQ ID NO: 3, respectively; the light chain amino acid sequences encoded by the light chain variable region cDNA sequences of 6A7E2 and 1H119 are shown in SEQ ID NO: 4 and SEQ ID NO: 5, respectively. The antigen complementarity determinant (CDR) sequence of 6A7E2 is shown in Table 1. The antigen complementarity determinant (CDR) sequence of 1H119 is shown in Table 2.

[0122] Table 1. Antigen complementarity determinant (CDR) sequence of 6A7E2.

[0123] Table 2. Antigen complementarity determinant (CDR) sequence of 1H119.

[0124] The method for detecting the binding activity with human TL1A protein is as follows: A 1 μg / mL human TL1A (TL1A-ECD, SEQ ID NO:1) protein solution was coated into a 96-well high-affinity plate at 100 μL / well and incubated overnight at 4°C with shaking. The next day, the plate was washed three times with 300 μL PBST (Tween 20: 0.5‰), then blocked with 100 μL / well of 5% BSA / PBS for 1 hour with shaking at room temperature. The plate was washed three times with 300 μL PBST. Samples were prepared with PBS. 100 μL of PBS was added to each well of a 96-well plate and incubated for 1 hour with shaking at room temperature. The plate was washed three times with 300 μL PBST. Secondary antibody goat anti-mouse IgG HRP solution (Thermo Fisher, catalog number A16090) was prepared and added to each well of a 96-well plate at 100 μL. The plate was incubated for 0.5 hours with shaking at room temperature. The plate was washed four times with 300 μL PBST. Add 100 μL / well TMB and develop for 3 minutes. Add 100 μL / well 0.6 N H2SO4 to stop the development and measure the OD. 450 nm.

[0125] Method for detecting the antibody's ability to block the binding of human TL1A to human DR3 protein: 100 μL / well of human DR3 (uniprot accession number: Q93038) was coated onto a 96-well high-affinity plate and incubated overnight at 4°C with shaking. The next day, the plate was washed three times with 300 μL of PBST (Tween 20: 0.5‰), then blocked with 100 μL / well of 5% BSA / PBS for 1 hour with shaking at room temperature. The plate was washed three times with 300 μL of PBST. 100 μL of the sample solution was premixed with biotin-labeled TL1A solution (labeling reagent purchased from Thermo Fisher, A35358) (2 μg / mL) at a 1:1 (v:v) ratio and incubated at room temperature for 0.5 hours. 100 μL / well of the mixture was added to the 96-well plate and incubated for 1 hour with shaking at room temperature. The plate was washed three times with 300 μL of PBST. HRP-labeled streptavidin solution (Beyotime, catalog number A0303, hereinafter the same) was added to 100 μL / well of a 96-well plate and shaken at room temperature for 0.5 hours. The plate was washed four times with 300 μL PBST. 100 μL / well of TMB was added, and the plate was incubated for 5 min. 100 μL / well of 0.6N H₂SO₄ was added to stop the incubation, and the OD was measured. 450 nm.

[0126] Example 2: Binding of chimeric antibodies Y10-L5 and Y10-L15 to human TL1A protein The variable regions of the heavy and light chains of mouse antibody 6A7E2 were ligated to the constant regions of the heavy and κ chains of human IgG1, respectively, to obtain the human-mouse chimeric antibody Y10-L5, whose heavy chain sequence is shown in SEQ ID NO:46 and light chain sequence is shown in SEQ ID NO:47. Similarly, the variable regions of the heavy and light chains of mouse antibody 1H119 were ligated to the constant regions of the heavy and κ chains of human IgG1, respectively, to obtain the human-mouse chimeric antibody Y10-L15, whose heavy chain sequence is shown in SEQ ID NO:48 and light chain sequence is shown in SEQ ID NO:49.

[0127] The binding activity of Y10-L5 and Y10-L15 to human TL1A protein was investigated. The detection method is described in Example 1 (the secondary antibody was changed to goat anti-human IgG HRP, Abcam, catalog number ab6858). The results are as follows. Figure 1 , 2 As shown, the EC50 values ​​for the binding of Y10-L5 and Y10-L15 to human TL1A protein are approximately 58.91 ng / mL and 54.16 ng / mL, respectively.

[0128] Example 3: Chimeric antibodies Y10-L5 and Y10-L15 block the binding of human TL1A to DR3 The activity of Y10-L5 in blocking the binding of human TL1A to human DR3 protein was investigated. The detection method is described in Example 1, and the results are as follows. Figure 3 As shown, the IC50 of Y10-L5 inhibiting the binding of human TL1A to DR3 protein is 487.9 ng / mL.

[0129] Assay for the activity of antibody in blocking the binding of human TL1A to CHO-DR3: CHO-DR3 cells were self-constructed CHO-K1 cells stably expressing human DR3. CHO-DR3 cells were digested and centrifuged, resuspended in PBS at a density of 2 x 10^6 cells / mL, and 100 μL was transferred to a centrifuge tube. The supernatant was discarded after centrifugation. Serial dilutions of the antibody sample were prepared using PBS. 100 μL of the antibody solution was premixed with biotin-labeled TL1A solution (2 μg / mL) at a 1:1 (v:v) ratio and incubated at room temperature for 0.5 hours. Then, 100 μL of the antigen-antibody mixture was added to the cells, and the mixture was incubated at 4°C with shaking for 1 hour. The cells were centrifuged, and the supernatant was discarded. APCstreptavidin flow cytometry antibody (BioLegend, catalog number 405207) was prepared. 400 μL of the secondary antibody solution was added to the cells, and the mixture was incubated at 4°C with shaking for 0.5 hours. Centrifuge cells at 1000 rpm for 3 minutes, discard the supernatant, and mix thoroughly with 400 μL PBS. Repeat twice. Transfer to a flow cytometer (Beckman, Cytoflex) for analysis. Flow cytometry results are shown below. Figure 4, 5 As shown, the IC50 values ​​of Y10-L5 and Y10-L15 for inhibiting the binding of human TL1A to CHO-DR3 were approximately 1.329 μg / mL and 1.476 μg / mL, respectively.

[0130] Example 4: Inhibition of TL1A-induced apoptosis in TF-1 cells by Y10-L5 TF-1 is a human hematologic malignancy cell line that naturally overexpresses human DR3. In TF-1 cells, TL1A acts on DR3, inducing Caspase 3 activation, which in turn leads to TF-1 cell apoptosis. The specific detection method was as follows: TF-1 cells were centrifuged, resuspended in 1640 medium (containing 10% FBS), and seeded into 96-well plates at a rate of 70,000 cells / well. Serially diluted antibody solution, as well as Cycloheximide (final concentration 50 μg / mL) and TL1A (TL1A-ECD, SEQ ID NO:1) solution (final concentration 0.1 μg / mL) were added. The mixture was incubated overnight at 37°C, and then analyzed using CellTiter-Glo (Promega, catalog number G7572). Results are as follows: Figure 6 As shown, the IC50 of Y10-L5 on TL1A-induced apoptosis in TF-1 cells was 0.3128 μg / mL.

[0131] Example 5: Inhibition of TL1A-induced NF-κB signaling pathway activation by Y10-L5 and Y10-L15 A Jurkat-NF-κB-DR3 reporter gene cell line overexpressing DR3 protein was constructed. The Jurkat-NF-κB-DR3 construction method is as follows: using Lipo 2000 (ThermoFisher, 11668019) liposome transfection, plasmids carrying the target genes DR3 and NF-κB-luc (Beyotime Biotechnology, D2204-100μg) were transfected into Jurkat cells. After dual screening using both resistance and target genes, Jurkat-NF-κB-DR3 reporter gene cells stably overexpressing DR3 and NF-κB-luc were finally obtained.

[0132] Jurkat-NF-κB-DR3 cells were centrifuged and resuspended in 1640 medium (containing 10% FBS) at a density of 1.0 x 10^6 cells / mL. After mixing, 10,000 cells were transferred to 384-well plates with white walls and clear bottoms. A serially diluted antibody sample solution was prepared by adding complete culture medium and TL1A (TL1A-ECD, SEQ ID NO:1) solution (final concentration 0.2 μg / mL). After incubation at 37°C for 5 h, ONE-Glo Luciferase detection solution (Promega, catalog number E6110) was added, and the reaction was allowed to proceed for 1-3 minutes. The cells were then transferred to a multi-plate reader (Tecan Spark 20M) for detection. Results are as follows: Figure 7 , 8 As shown, the IC50 values ​​of Y10-L5 and Y10-L15 for inhibiting TL1A-induced NF-κB signaling pathway activation were 0.1023 μg / mL and 0.1498 μg / mL, respectively.

[0133] Example 6: Binding of humanized antibody to human TL1A protein Humanization of chimeric antibodies Y10-L5 and Y10-L15 was achieved using the CDR transplantation method, yielding humanized antibodies Y10-L5VH1-1VL1-1 and Y10-L15VH1VL1. Furthermore, Y10-L5VH1-1VL1-1 and Y10-L15VH1VL1 underwent FC mutation modification, resulting in Y10-L5VH1-1VL1-1 (YTE), Y10-L5VH1-1VL1-1 (LS), Y10-L15VH1VL1 (YTE), and Y10-L15VH1VL1 (LS). The sequences of these antibodies are shown in Table 3 below.

[0134] Table 3. Antibody sequences of humanized antibodies

[0135] The binding activity of the above-mentioned humanized antibody to human TL1A protein was studied, and the detection method is described in Example 2. The detection results are as follows: Figure 9 , 10 As shown: the EC50 values ​​for binding to human TL1A protein by Y10-L5VH1-1VL1-1, Y10-L5VH1-1VL1-1 (YTE), and Y10-L5VH1-1VL1-1 (LS) were 100.3 ng / mL, 289.8 ng / mL, and 96.34 ng / mL, respectively; the EC50 values ​​for binding to human TL1A protein by Y10-L15VH1VL1, Y10-L15VH1VL1 (YTE), and Y10-L15VH1VL1 (LS) were 122.5 ng / mL, 73.9 ng / mL, and 24.93 ng / mL, respectively.

[0136] Example 7: Binding of humanized antibody to FcRn receptor The binding affinity of humanized antibodies (Y10-L5VH1-1VL1-1, Y10-L5VH1-1VL1-1(YTE), Y10-L5VH1-1VL1-1(LS), Y10-L15VH1VL1, Y10-L15VH1VL1(YTE), Y10-L15VH1VL1(LS)) to FcRn receptors was detected using the TR-FRET method. (Novozymes, Add&Read Human Fc Kit, catalog number DD2102-01) Figure 11 , 12 As shown: Y10-L5VH1-1VL1-1(YTE) and Y10-L5VH1-1VL1-1(LS) have significantly stronger binding affinity to human FcRn than Y10-L5VH1-1VL1-1, and Y10-L15VH1VL1(YTE) and Y10-L15VH1VL1(LS) have significantly stronger binding affinity to human FcRn than Y10-L15VH1VL1.

[0137] Experimental conclusion: The Fc-terminal mutation of the antibody in this invention enhances the binding of the antibody to the FcRn receptor.

[0138] Example 8: Humanized antibody blocks the binding of human TL1A to DR3 The activity of the humanized antibody in blocking the binding of human TL1A to CHO-DR3 was investigated; the detection method is described in Example 3. The results are as follows: Figure 13 , 14 As shown, the IC50 values ​​of Y10-L5VH1-1VL1-1, Y10-L5VH1-1VL1-1(YTE), and Y10-L5VH1-1VL1-1(LS) for inhibiting the binding of human TL1A to CHO-DR3 were approximately 0.7039 μg / mL, 1.333 μg / mL, and 0.7867 μg / mL, respectively. The IC50 values ​​of Y10-L15VH1VL1, Y10-L15VH1VL1(YTE), and Y10-L15VH1VL1(LS) for inhibiting the binding of human TL1A to CHO-DR3 were approximately 0.9727 μg / mL, 0.3376 μg / mL, and 0.5448 μg / mL, respectively.

[0139] Experimental conclusion: The antibody of this invention can block the binding of TL1A to its receptor DR3.

[0140] Example 9: Differential Scanning Fluorescence (DSF) Detection of Thermal Stability of Humanized Antibodies Dilute the test sample to 1 mg / mL with PBS. Dilute SYPRO Orange dye (MERCK, catalog number S5692) 100-fold with PBS. Add 1 μL of SYPRO Orange dye to 49 μL of the test antibody in a white PCR (Biorad) tube, set up replicates, pipette once, centrifuge, and place on ice. Run the pre-programmed procedure on the Biorad (CFX connect) to analyze and process the data. Results are as follows: Figure 15 , 16 As shown: the Tm1 of Y10-L5VH1-1VL1-1 and Y10-L15VH1VL1 are approximately 70.25℃ and 67.25℃, respectively. The Tm1 of Y10-L5VH1-1VL1-1(YTE), Y10-L5VH1-1VL1-1(LS), Y10-L15VH1VL1(YTE), and Y10-L15VH1VL1(LS) are approximately 58.75℃, 67.25℃, 59.25℃, and 65.25℃, respectively.

[0141] Experimental conclusion: The antibody of this invention has good thermal stability.

[0142] Example 10: Inhibition of TL1A-induced apoptosis in TF-1 cells by humanized antibodies The effect of humanized antibodies on TL1A-induced apoptosis in TF-1 cells was investigated; the detection method is described in Example 4. Results are as follows: Figure 17 , 18 As shown in Figure 19, the IC50 values ​​of Y10-L5VH1-1VL1-1 and Y10-L15VH1VL1 against TL1A-induced apoptosis in TF-1 cells were approximately 0.09571 μg / mL and 0.1113 μg / mL, respectively. The IC50 values ​​of Y10-L5VH1-1VL1-1 (YTE), Y10-L5VH1-1VL1-1 (LS), Y10-L15VH1VL1 (YTE), and Y10-L15VH1VL1 (LS) against TL1A-induced apoptosis in TF-1 cells were approximately 0.1878 μg / mL, 0.2483 μg / mL, 0.1019 μg / mL, and 0.1449 μg / mL, respectively.

[0143] Experimental conclusion: The antibody of this invention can effectively block TL1A-induced apoptosis in TF-1 cells.

[0144] Example 11: Inhibition of TL1A-induced NF-κB signaling pathway activation by humanized antibodies The activation of the TL1A-induced NF-κB signaling pathway by humanized antibodies was investigated; the detection method is described in Example 5. Results are as follows: Figure 20 , 21As shown, the IC50 values ​​of Y10-L5VH1-1VL1-1, Y10-L5VH1-1VL1-1 (YTE), and Y10-L5VH1-1VL1-1 (LS) for inhibiting TL1A-induced NF-κB signaling pathway activation were approximately 0.1270 μg / mL, 0.3696 μg / mL, and 0.1280 μg / mL, respectively. The IC50 values ​​of Y10-L15VH1VL1, Y10-L15VH1VL1 (YTE), and Y10-L15VH1VL1 (LS) for inhibiting TL1A-induced NF-AL signaling pathway activation were approximately 24.50 ng / mL, 7.862 ng / mL, and 28.63 ng / mL, respectively.

[0145] Experimental conclusion: The antibody of this invention can effectively block TL1A-induced activation of the NF-κB signaling pathway.

[0146] Example 12: Inhibitory effect of humanized antibody on TL1A-induced expression of IFN-γ in human blood cells The inhibitory effect of humanized antibodies on IFN-γ expression induced by recombinant TL1A in human PBMCs was investigated. The detection method was as follows: PBMCs were resuscitated one day in advance and cultured in 1640 medium (containing 10% FBS). PBMCs were seeded into 96-well round-bottom plates, with 100,000 cells per well. A mixture of recombinant TL1A (TL1A-ECD, SEQ ID NO:1, final concentration 300 ng / mL), IL-12 (Peprotech, 200-12-10), and IL-18 (RD, 9124-IL-050 / CF) (final concentrations 1 ng / mL IL-12 and 5 ng / mL IL-18) cytokines was added to the cells. The serially diluted test antibody was added and mixed well. The mixture was incubated for 24 h, centrifuged at 600 x g for 10 min, and the supernatant was collected. IFN-γ expression was detected by ELISA (BD, catalog number 555142). The results are as follows: Figure 22 As shown, the IC50 values ​​of Y10-L5VH1-1VL1-1(LS) and Y10-L15VH1VL1(LS) for inhibiting IFN-γ expression induced by recombinant TL1A in human PBMCs were approximately 1.079 μg / mL and 0.8435 μg / mL, respectively.

[0147] The inhibitory effect of humanized antibodies on the expression of IFN-γ in whole blood cells stimulated by immune complexes was investigated. The detection method was as follows: One day in advance, 50 μL of 0.5 mg / mL IgG (Jackson ImmunoResearch, catalog number 009-000-003) was coated into 96-well cell culture plates and incubated overnight at 4°C. The cell culture plates were washed with PBS, and 50 μL of 0.04 mg / mL mouse anti-human IgG (Jackson ImmunoResearch, catalog number 209-005-082) was added to each well, and incubated at 37°C for 2 hours. The cell culture plates were washed with PBS, and 150 μL of freshly drawn heparinized whole blood from volunteers was added, along with 25 μL of a cytokine mixture of IL-12 and IL-18 (final concentration 1 ng / mL IL-12 + 5 ng / mL IL-18). Add serially diluted test antibody, mix well, incubate for 24 h, centrifuge at 600 x g for 10 min to collect the supernatant, and detect IFN-γ expression using an ELISA kit (BD, catalog number 555142). Results are as follows: Figure 23 , 24 As shown, the IC50 values ​​of Y10-L5VH1-1VL1-1(LS) and Y10-L5VH1-1VL1-1(YTE) for inhibiting the expression of IFN-γ in whole blood cells by immune complex stimulation were approximately 1.56 nM and 2.86 nM, respectively, while the IC50 value of Y10-L15VH1VL1(LS) for inhibiting the expression of IFN-γ in whole blood cells by immune complex stimulation was approximately 0.74 nM.

[0148] Experimental conclusion: The antibody of this invention can effectively inhibit the secretion of IFN-γ by blood cells.

[0149] Example 13: Binding of humanized antibodies to mouse TL1A, rat TL1A, and cynomolgus monkey TL1A The binding activity of the humanized antibody with mouse TL1A, rat TL1A, and cynomolgus monkey TL1A (all purchased from ARCOBio, M5243, R5249, and C5241) was studied. The detection method is described in Example 2. The results are as follows: Figure 25 , 26 As shown in Figure 27, Y10-L5VH1-1VL1-1 exhibits strong binding activity with TL1A in mice, cynomolgus monkeys, and rats, while Y10-L15VH1VL1 does not bind to TL1A in mice or rats, but exhibits strong binding activity with TL1A in cynomolgus monkeys. Y10-L5VH1-1VL1-1 (YTE) and Y10-L5VH1-1VL1-1 (LS) both show strong binding activity with TL1A in rats.

[0150] Experimental conclusion: The antibody of this invention can bind to TL1A protein in mice, rats and cynomolgus monkeys.

[0151] Example 14: Detection of antibody affinity The antigen-antibody affinity was detected using the biomembrane interference (BLI) method. Detection conditions: temperature set at 30℃, sample plate rotation speed at 1000 rpm. Human TL1A-his (TL1A-ECD, SEQ ID NO:1) protein was diluted with HBS-EP solution. The antibody to be tested was captured using an AHC2 chip (capture time 300 seconds). The chip was then immersed in antigen solutions of different concentrations to detect the binding constant (ka). Subsequently, the chip was immersed in HBS-EP to detect the dissociation constant (kd). The detection results are shown in Table 4. The affinities of Y10-L5, Y10-L5VH1-1VL1-1, and Y10-L15VH1VL1 with human TL1A-his were approximately 0.236 nM, 0.0893 nM, and 0.1603 nM, respectively.

[0152] The surface plasmon resonance (SPR) method was used to detect antigen-antibody affinity. Using mouse TL1A-his (ARCOBio, M5243) as the antigen, a certain concentration of antibody was incubated with a protein A sensor chip (Cytiva, 29127556) for antibody capture. During the antigen binding phase, TL1A-his protein was used as the mobile phase to bind with the antibody captured on the sensor chip. During the dissociation phase, continuous elution was performed with HBS-EP buffer. The binding of the antibody to mouse TL1A-his on the sensor chip was quantitatively detected using a Biacore 8k (GE Healthcare). The results are shown in Table 5. The affinity of Y10-L5VH1-1VL1-1 for mouse TL1A-his was 0.0385 nM.

[0153] Experimental conclusion: The antibody of this invention has a strong affinity for TL1A protein.

[0154] Table 4. Affinity of anti-human TL1A antibody to human TL1A-his

[0155] Table 5. Affinity of anti-human TL1A antibody to mouse TL1A-his

[0156] Example 15: Pharmacological study of humanized antibody in a TNBS-induced mouse enteritis model An acute enteritis model was induced in 8-12 week old male C57BL / 6 mice by colonic instillation of 2.5% TNBS (Meilunbio) solution (50% ethanol). On the day of modeling and on day 3 post-modeling, mice were intraperitoneally injected with 30 mg / kg of Y10-L5VH1-1VL1-1, Y10-L5VH1-1VL1-1 (LS), and C320 (positive control, TL1A antibody, Duvakitug, US10822422B2, Taizhou Baiying Biotechnology Expression). On day 6 post-modeling, the experimental mice were euthanized. The colon and rectum of the mice were harvested, their length measured, and histopathological examination performed. The colon and rectum tissue was fixed and sent for HE staining examination, and pathological scoring was performed by a pathologist according to the following scoring table.

[0157] The results are as follows Figure 28 , 29 As shown, L5VH1-1VL1-1 and L5VH1-1VL1-1 (LS) significantly inhibited TNBS-induced colorectal inflammation; histological scores showed that, compared with the model group, the animals in the L5VH1-1VL1-1 and L5VH1-1VL1-1 (LS) treatment groups had less damage to the intestinal mucosal structure.

[0158] Example 16: Pharmacokinetic Study of Humanized Antibody in hFcRn Transgenic Mice Female B-hFcRn C57BL / 6 mice (Biocytogen) were intraperitoneally injected with 5 mg / kg of Y10-L5VH1-1VL1-1 (YTE), Y10-L5VH1-1VL1-1 (LS), Y10-L15VH1VL1 (YTE), Y10-L15VH1VL1 (LS), or PM219 (positive control, TL1A antibody, Tulisokibart, US11999789B, Taizhou Baiying Biotechnology Expression). Three mice were in each group. Serum samples were collected at 0.5 h, 4 h, 24 h, 48 h, 72 h, 96 h, 168 h, 240 h, 336 h, 408 h, 504 h, and 696 h post-drug administration. Serum drug concentrations were detected using ELISA. The detection method is as follows: A 1 μg / mL solution of human TL1A protein (TL1A-ECD, SEQ ID NO:1) was coated onto a 96-well high-affinity plate at 100 μL / well and incubated overnight at 4°C with shaking. The next day, the plate was washed three times with 300 μL PBST (Tween 20: 0.5‰), then blocked with 100 μL / well of 5% BSA / PBS for 2 hours with shaking at room temperature. The plate was then washed three times with 300 μL PBST. Serum samples were diluted a certain factor with blank serum, and then further diluted to a 1% serum concentration with PBS. 100 μL / well of the solution was added to a 96-well plate, and the plate was incubated at room temperature with shaking for 1 hour. The plate was then washed three times with 300 μL PBST. Secondary antibody (goat anti-human IgG HRP, Abcam, catalog number ab6858) was prepared and added to a 96-well plate at 100 μL / well with shaking at room temperature for 30 minutes. The plate was then washed four times with 300 μL PBST. Add 100 μL / well TMB and develop color for 3 minutes. Add 100 μL / well 0.6 NH₂SO₄ to stop the color development and measure the OD. 450 nm. Simultaneously, a standard curve for the antibody to be tested was prepared using serum, and detection was performed according to the above method. The detection values ​​from the serum sample were substituted into the standard curve for calculation to obtain the drug concentration in the sample.

[0159] The results are as follows Figure 30 As shown in Table 6.

[0160] Experimental conclusion: In the hFcRn mouse model, the antibody of this invention exhibits good pharmacokinetic properties.

[0161] Table 6. Pharmacokinetic parameters of the antibody in hFcRn mice

[0162] Example 17: Pharmacokinetic Study of Humanized Antibody in Cynomolgus Monkey Model Cynomolgus monkeys (n=2, one male and one female) were administered Y10-L5VH1-1VL1-1 (YTE) and Y10-L5VH1-1VL1-1 (LS) at 5 mg / kg intravenously. Serum samples were collected at 0.5 h, 1 h, 2 h, 6 h, 24 h, 48 h, 96 h, 168 h, 336 h, 504 h, and 672 h post-administration. Serum drug concentrations at different time points were detected by ELISA. Pharmacokinetic parameters are shown in Table 7.

[0163] Experimental conclusion: In the cynomolgus monkey model, the antibody of this invention exhibits good pharmacokinetic properties.

[0164] Table 7. Pharmacokinetic parameters of the antibody in cynomolgus monkeys

[0165] Example 18: Pharmacological study of humanized antibody in a TNBS-induced hTL1A mouse enteritis model An acute enteritis model was induced in 8-12 week old TL1A humanized male C57BL / 6 mice via colonic perfusion with 1.5% TNBS (Meilunbio) solution (50% ethanol) (Day 0). On Day-1, Day 1, and Day 3, mice were intraperitoneally injected with 30 mg / kg of Y10-L15VH1VL1 (LS), Y10-L5VH1-1VL1-1 (LS), and PM219 (Taizhou Baiying Biotechnology Expression). Animal weight and DAI scores were recorded daily during the experiment. On day 5 post-modeling, the mice were euthanized. The colon and rectum were harvested, length measured, and inflammatory factors detected. The colon and rectum tissue was fixed and sent for HE staining examination, and pathological scoring was performed by a pathologist according to the following scoring table.

[0166] The results are as follows Figure 31 , 32 As shown in Figure 33: Body weight and DAI scores showed that, compared with the model group (Vehicle), Y10-L15VH1VL1 (LS) and Y10-L5VH1-1VL1-1 (LS) significantly increased animal body weight and alleviated disease severity; inflammatory factor detection showed that, compared with the model group, animals in the Y10-L15VH1VL1 (LS) and Y10-L5VH1-1VL1-1 (LS) treatment groups had milder intestinal inflammation.

[0167] Experimental conclusion: In the TNBS-induced hTL1A mouse enteritis model, the antibody of this invention has good efficacy against the mouse inflammatory enteritis model.

[0168] The technical solutions of the present invention are not limited to the specific embodiments described above. Any technical modifications made in accordance with the technical solutions of the present invention fall within the protection scope of the present invention.

Claims

1. A TL1A antibody or an antigen-binding fragment thereof, wherein the TL1A antibody or the antigen-binding fragment thereof comprises a complementarity-determining region (CDR) comprising: a1) having VH-CDR1, VH-CDR2, and VH-CDR3 contained in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO: 2; and / or, having VL-CDR1, VL-CDR2, and VL-CDR3 contained in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO: 4; or a2) VH-CDR1, VH-CDR2, and VH-CDR3 contained in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO: 3; and / or, VL-CDR1, VL-CDR2, and VL-CDR3 contained in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO: 5; or a3) Any amino acid sequence in the CDR sequence of a1) or a2) above may optionally include an amino acid sequence that has been added, deleted, modified and / or substituted with 1-3 amino acid residues and retains TL1A binding activity. in, The CDR is defined by a definition scheme of Kabat, Chothia, IMGT, Martin, Contact, AbM, or a combination thereof.

2. The TL1A antibody or its antigen-binding fragment according to claim 1, characterized in that, The TL1A antibody or its antigen-binding fragment includes: b1) Heavy chain variable regions, including VH-CDR1, VH-CDR2, and VH-CDR3; and The light chain variable region includes VL-CDR1, VL-CDR2 and VL-CDR3; The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, and VL-CDR3 are shown in SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10, respectively. The amino acid sequence of VL-CDR2 is YTS, and the CDR is defined according to the IMGT definition scheme; or The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are shown in SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 10, respectively. The CDRs are defined using the Kabat scheme; or The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are shown in SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 10, respectively, and the CDRs are defined according to the Chothia scheme; or The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are shown in SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, and SEQ ID NO: 23, respectively. The CDRs are defined using the Contact definition scheme; or The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are shown in SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 10, respectively, and the CDRs are defined using the AbM definition scheme; or b2) Heavy chain variable regions, including VH-CDR1, VH-CDR2, and VH-CDR3; and The light chain variable region includes VL-CDR1, VL-CDR2 and VL-CDR3; The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, and VL-CDR3 are shown in SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, and SEQ ID NO: 30, respectively. The amino acid sequence of VL-CDR2 is YSS, and the CDR is defined according to the IMGT scheme; or The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are shown in SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 30, respectively, and the CDRs are defined according to the Kabat scheme; or The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are shown in SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 30, respectively, and the CDRs are defined according to the Chothia scheme; or The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are shown in SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, and SEQ ID NO: 43, respectively. The CDRs are defined using the Contact definition scheme; or The amino acid sequences of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3 are shown in SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 30, respectively, and the CDRs are defined using the AbM definition scheme; or b3) Heavy chain variable region; and Light chain variable region, Wherein, compared with any of the heavy chain variable regions and / or light chain variable regions described in b1)-b2), at least one CDR contains a mutation, wherein the mutation is a substitution, deletion or addition of one or more amino acids; preferably, the substitution is a conservative substitution.

3. The TL1A antibody or its antigen-binding fragment according to any one of claims 1-2, characterized in that, The heavy chain variable region of the TL1A antibody or its antigen-binding fragment also includes the framework region of the heavy chain variable region; Preferably, the framework region of the heavy chain variable region includes the framework region of the heavy chain variable region or a mutant thereof derived from immunoglobulins of mice, primates, cattle, horses, pigs, sheep, goats, dogs, cats, rabbits, camels, donkeys, deer, minks, chickens, ducks, or geese. More preferably, it includes the framework region of the heavy chain variable region or a mutant thereof derived from immunoglobulins of mice, rats, lemurs, macaques, chimpanzees, or humans. More preferably, it includes the framework region of the heavy chain variable region or a mutant thereof derived from immunoglobulins of humans or mice. Even more preferably, it includes the framework region of the heavy chain variable region or a mutant thereof derived from immunoglobulins of humans. Preferably, the light chain variable region of the TL1A antibody or its antigen-binding fragment further includes the framework region of the light chain variable region; Preferably, the framework region of the light chain variable region includes the framework region of the light chain variable region or a mutant thereof derived from immunoglobulins of mice, primates, cattle, horses, pigs, sheep, goats, dogs, cats, rabbits, camels, donkeys, deer, minks, chickens, ducks, or geese. More preferably, it includes the framework region of the light chain variable region or a mutant thereof derived from immunoglobulins of mice, rats, lemurs, macaques, chimpanzees, or humans. More preferably, it includes the framework region of the light chain variable region or a mutant thereof derived from immunoglobulins of humans or mice. Even more preferably, it includes the framework region of the light chain variable region or a mutant thereof derived from immunoglobulins of humans.

4. The TL1A antibody or its antigen-binding fragment according to any one of claims 1-3, characterized in that, The TL1A antibody or its antigen-binding fragment includes: The heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 2, 3, 50 or SEQ ID NO: 56, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity with it; and / or, the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 4, 5, 51 or SEQ ID NO: 57, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity with it; Preferably, the antibody or its antigen-binding fragment comprises: 1) A heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 2, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 4, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; 2) Heavy chain variable region, comprising an amino acid sequence as shown in SEQ ID NO: 3, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, light chain variable region, comprising an amino acid sequence as shown in SEQ ID NO: 5, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; 3) Heavy chain variable region, comprising an amino acid sequence as shown in SEQ ID NO: 50, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, light chain variable region, comprising an amino acid sequence as shown in SEQ ID NO: 51, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; 4) Heavy chain variable region, comprising an amino acid sequence as shown in SEQ ID NO: 56, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, light chain variable region, comprising an amino acid sequence as shown in SEQ ID NO: 57, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

5. The TL1A antibody or its antigen-binding fragment according to any one of claims 1-4, characterized in that, The TL1A antibody or its antigen-binding fragment further includes a heavy chain constant region and / or a light chain constant region; Preferably, the heavy chain constant region includes the heavy chain constant region or its mutant from immunoglobulins derived from mice, primates, cattle, horses, pigs, sheep, goats, dogs, cats, rabbits, camels, donkeys, deer, minks, chickens, ducks, or geese; more preferably, it includes the heavy chain constant region or its mutant from immunoglobulins derived from mice, rats, lemurs, macaques, chimpanzees, or humans; even more preferably, it includes the heavy chain constant region or its mutant from immunoglobulins derived from humans or mice; and even more preferably, it includes the heavy chain constant region or its mutant from immunoglobulins derived from humans. Preferably, the light chain constant region includes the light chain constant region or its mutant from immunoglobulins derived from mice, primates, cattle, horses, pigs, sheep, goats, dogs, cats, rabbits, camels, donkeys, deer, minks, chickens, ducks, or geese; more preferably, it includes the light chain constant region or its mutant from immunoglobulins derived from mice, rats, lemurs, macaques, chimpanzees, or humans; even more preferably, it includes the light chain constant region or its mutant from immunoglobulins derived from humans or mice; and even more preferably, it includes the light chain constant region or its mutant from immunoglobulins derived from humans. Preferably, the heavy chain constant region includes a heavy chain constant region derived from IgA1, IgA2, IgD, IgE, IgG1, IgG2, IgG3, IgG4 or IgM immunoglobulin; Preferably, the heavy chain constant region includes the human IgG1 heavy chain constant region or a mutant thereof. More preferably, the mutant of the human IgG1 heavy chain constant region includes: 1) having the following substitutions compared to its derived wild-type sequence: M252Y, S254T and / or T256E; and / or, 2) having the following substitutions compared to its derived wild-type sequence: M428L and / or N434S, wherein the amino acid positions mentioned above are positions according to the EU numbering system; Preferably, the light chain constant region includes light chain constant regions derived from κ and λ immunoglobulins; Preferably, the heavy chain constant region comprises an amino acid sequence as shown in SEQ ID NO: 52, 54 or 55, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity with it; Preferably, the light chain constant region comprises an amino acid sequence as shown in SEQ ID NO: 53, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

6. The TL1A antibody or its antigen-binding fragment according to any one of claims 1-5, characterized in that, The TL1A antibody or its antigen-binding fragment is selected from the Fab fragment, Fab' fragment, F(ab')2 fragment, Fv fragment, single-chain Fv (scFv), dsFv and Fd fragment; Preferably, the TL1A antibody or its antigen-binding fragment is selected from murine antibodies, chimeric antibodies, humanized antibodies, and fully human antibodies; Preferably, the TL1A antibody or its antigen-binding fragment comprises a heavy chain and / or a light chain, wherein, q1) The heavy chain comprises amino acids 19-472 of SEQ ID NO: 46, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, the light chain comprises amino acids 19-232 of SEQ ID NO: 47, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; or q2) The heavy chain comprises amino acids 19-473 of SEQ ID NO: 48, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, the light chain comprises amino acids 19-232 of SEQ ID NO: 49, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; or q3) The heavy chain comprises the amino acid sequence shown in SEQ ID NO: 58, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, the light chain comprises the amino acid sequence shown in SEQ ID NO: 59, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; or q4) The heavy chain comprises the amino acid sequence shown in SEQ ID NO: 60, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, the light chain comprises the amino acid sequence shown in SEQ ID NO: 61, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; or q5) The heavy chain comprises the amino acid sequence shown in SEQ ID NO: 62, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, the light chain comprises the amino acid sequence shown in SEQ ID NO: 63, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; or q6) The heavy chain comprises the amino acid sequence shown in SEQ ID NO: 64, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, the light chain comprises the amino acid sequence shown in SEQ ID NO: 65, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; or q7) The heavy chain comprises the amino acid sequence shown in SEQ ID NO: 66, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, the light chain comprises the amino acid sequence shown in SEQ ID NO: 67, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; or q8) The heavy chain comprises an amino acid sequence as shown in SEQ ID NO: 68, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, the light chain comprises an amino acid sequence as shown in SEQ ID NO: 69, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

7. A chimeric antigen receptor, characterized in that, The chimeric antigen receptor includes an antigen-binding domain, a transmembrane domain, and an intracellular signal transduction domain, wherein the antigen-binding domain includes the TL1A antibody or its antigen-binding fragment as described in any one of claims 1-6.

8. An isolated nucleic acid molecule, characterized in that, The nucleic acid molecule encodes the TL1A antibody or its antigen-binding fragment as described in any one of claims 1-6, or the chimeric antigen receptor as described in claim 7.

9. A carrier, characterized in that, The carrier comprises the nucleic acid molecule of claim 8. Preferably, the carrier is an expression carrier.

10. A cell, characterized in that, The cell comprises the TL1A antibody or its antigen-binding fragment as described in any one of claims 1-6, the chimeric antigen receptor as described in claim 7, the nucleic acid molecule as described in claim 8, or the vector as described in claim 9.

11. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises the TL1A antibody or its antigen-binding fragment as described in any one of claims 1-6, the chimeric antigen receptor as described in claim 7, the nucleic acid molecule as described in claim 8, the vector as described in claim 9, or the cell as described in claim 10.

12. The pharmaceutical composition according to claim 11, characterized in that, The pharmaceutical composition also includes a pharmaceutically acceptable carrier; Preferably, the pharmaceutical composition is used for the prevention or treatment of diseases related to TL1A; Preferably, the TL1A-related diseases include fibrotic diseases, as well as inflammatory diseases or autoimmune diseases; Preferably, the inflammatory disease or autoimmune disease includes asthma, arthritis, multiple sclerosis, inflammatory bowel disease, chronic obstructive pulmonary disease, glomerulonephritis, psoriasis, allergic dermatitis, bronchitis, allergic rhinitis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, and systemic sclerosis-associated interstitial lung disease; Preferably, the inflammatory bowel disease includes ulcerative colitis and Crohn's disease; Preferably, the fibrotic diseases include systemic fibrotic diseases and organ-specific fibrotic diseases; Preferably, the systemic fibrotic diseases include systemic sclerosis, multifocal fibrosis, renal systemic fibrosis, and scleroderma graft-versus-host disease; Preferably, the organ-specific fibrotic diseases include pulmonary fibrosis, cardiac fibrosis, renal fibrosis, hepatic and portal vein fibrosis, radiation-induced fibrosis, bladder fibrosis, intestinal fibrosis, peritoneal sclerosis, diffuse fasciitis, and scar formation.

13. A product characterized in that, The product includes the TL1A antibody or its antigen-binding fragment as described in any one of claims 1-6, and the product includes at least one of reagents, chips, test strips and detection kits.

14. The product according to claim 13, characterized in that, The product can be used for: 1) Detect the presence or content of TL1A in the sample; or 2) Diagnose or assess the prognosis of diseases related to TL1A; Preferably, the TL1A-related diseases include fibrotic diseases, as well as inflammatory diseases or autoimmune diseases; Preferably, the inflammatory disease or autoimmune disease includes asthma, arthritis, multiple sclerosis, inflammatory bowel disease, chronic obstructive pulmonary disease, glomerulonephritis, psoriasis, allergic dermatitis, bronchitis, allergic rhinitis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, and systemic sclerosis-associated interstitial lung disease; Preferably, the inflammatory bowel disease includes ulcerative colitis and Crohn's disease; Preferably, the fibrotic diseases include systemic fibrotic diseases and organ-specific fibrotic diseases; Preferably, the systemic fibrotic diseases include systemic sclerosis, multifocal fibrosis, renal systemic fibrosis, and scleroderma graft-versus-host disease; Preferably, the organ-specific fibrotic diseases include pulmonary fibrosis, cardiac fibrosis, renal fibrosis, hepatic and portal vein fibrosis, radiation-induced fibrosis, bladder fibrosis, intestinal fibrosis, peritoneal sclerosis, diffuse fasciitis, and scar formation; Preferably, the antibody or its antigen-binding fragment is conjugated to a detectable marker; Preferably, the detectable marker is a fluorescent or luminescent marker, more preferably selected from any one of acridine ester, acridine sulfonamide, luminol, isoluminol, horseradish peroxidase, and alkaline phosphatase.

15. Use of the TL1A antibody or antigen-binding fragment thereof according to any one of claims 1-6, the chimeric antigen receptor according to claim 7, the nucleic acid molecule according to claim 8, the vector according to claim 9, the cell according to claim 10, or the pharmaceutical composition according to any one of claims 11-12 in the preparation of a medicament for the prevention or treatment of TL1A-related diseases.

16. The use according to claim 15, characterized in that, The diseases associated with TL1A include fibrotic diseases, as well as inflammatory diseases or autoimmune diseases; Preferably, the inflammatory disease or autoimmune disease includes asthma, arthritis, multiple sclerosis, inflammatory bowel disease, chronic obstructive pulmonary disease, glomerulonephritis, psoriasis, allergic dermatitis, bronchitis, allergic rhinitis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, and systemic sclerosis-associated interstitial lung disease; Preferably, the inflammatory bowel disease includes ulcerative colitis and Crohn's disease; Preferably, the fibrotic diseases include systemic fibrotic diseases and organ-specific fibrotic diseases; Preferably, the systemic fibrotic diseases include systemic sclerosis, multifocal fibrosis, renal systemic fibrosis, and scleroderma graft-versus-host disease; Preferably, the organ-specific fibrotic diseases include pulmonary fibrosis, cardiac fibrosis, renal fibrosis, hepatic and portal vein fibrosis, radiation-induced fibrosis, bladder fibrosis, intestinal fibrosis, peritoneal sclerosis, diffuse fasciitis, and scar formation.

17. A method for detecting the presence or content of TL1A in a sample, wherein the sample is contacted with the TL1A antibody or antigen-binding fragment of any one of claims 1-6, and the formation of the complex is detected, provided that the TL1A antibody or antigen-binding fragment of any one of claims 1-6 is allowed to form a complex with TL1A. The method is not for diagnostic or therapeutic purposes.

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