Monoclonal antibody specifically binding to TNF-like ligand 1A (TL1A) or antigen-binding fragment thereof, and use thereof

Abstract

The present invention relates to the field of biotechnology and medicine, in particular to a monoclonal antibody or an antigen binding fragment thereof that specifically binds to TNF-like ligand 1A (TL1A). The invention further relates to nucleic acids encoding said antibodies, expression vectors, host cells and methods for the production thereof, methods for the production of the antibodies according to the invention, pharmaceutical compositions comprising the antibodies according to the invention or antigen-binding fragments thereof, and methods for the production of the antibodies according to the invention. As well as pharmaceutical compositions comprising the antibodies or antigen binding fragments thereof according to the invention and other therapeutically active compounds; to a method for the treatment of a TL1A-mediated disease or disorder, to the use of said antibody or antigen binding fragment thereof or pharmaceutical composition thereof for the treatment of a TL1A-mediated disease or disorder, and to the use of said antibodies or antigen binding fragments thereof and other therapeutically active compounds according to the invention for the treatment of TL1A-mediated diseases or disorders.

Description

Technical Field

[0001] The present invention relates to the fields of biotechnology and medicine, and in particular to monoclonal antibodies or antigen-binding fragments thereof that specifically bind to TNF-like ligand 1A (TL1A). The present invention further relates to nucleic acids encoding the antibodies, expression vectors, host cells, and methods for producing the same, methods for producing the antibodies according to the invention, pharmaceutical compositions comprising the antibodies according to the invention, pharmaceutical compositions comprising the antibodies according to the invention and other therapeutically active compounds, methods for treating TNF-like ligand 1A (TL1A)-mediated diseases or conditions, uses of the antibodies or pharmaceutical compositions thereof for treating TL1A-mediated diseases or conditions, and uses of the antibodies and other therapeutically active compounds for treating TL1A-mediated diseases or conditions. Background Art Monoclonal antibodies in the form of chimeric, humanized or fully human molecules have been shown to be useful as effective drugs for the treatment of a variety of conditions and diseases.

[0002] TL1A (TNF-like ligand 1A) protein, also known as TNFSF15 (TNF superfamily member 15) and VEGI (vascular endothelial growth inhibitor), is a proinflammatory cytokine in a biologically active soluble trimeric form with a molecular weight of approximately 61.5 kDa, a member of the tumor necrosis factor family.

[0003] TL1A is initially expressed as a 251-amino acid transmembrane protein with a molecular weight of approximately 28.1 kDa (https: / / www.uniprot.org / uniprotkb / O95150 / entry). The general structure of the TL1A monomer consists of three topological domains: an unstructured cytoplasmic domain (35 AARs), a helical transmembrane region (21 AARs), and an extracellular C-terminal region (195 AARs). Under the influence of various factors, TL1A can be cleaved from the membrane by the ADAM17 protease (also known as TACE (TNFα-converting enzyme)); further, the polypeptide chain is cleaved between alanine residue 71 and leucine residue 72 to form a monomer of 180 amino acid residues (Zhai, Y., Ni, J., Jiang, G.-W., Lu, J., Xing, L., Lincoln, C., Carter, KC, Janat, F., Kozak, D., Xu, S., Rojas, L., Aggarwal, B.B., Ruben, S., Li, L.-Y., Gentz, R., Yu, G.-L. VEGI, a novel cytokine of the tumor necrosis factor family, is anangiogenesis inhibitor that suppresses the growth of colon carcinomas in vivo. (1999) FASEB J.13,181-189,doi:10.1096 / fasebj.13.1.181).

[0004] There is a prominent hydrophobic site on the surface of the native TL1A monomer that leads to the formation of a biologically active trimer (Zhan C., Yan Q., Patskovsky Yu., Li Z., Toro R., Meyer A., ​​Cheng H., Brenowitz M., Nathenson S.G., Almo SC, Biochemical and Structural Characterization of the Human TL1A Epidomain. (2009) Biochemistry: 48(32): 7636-7645, doi: 10.1021 / bi900031w).

[0005] TL1A is expressed by monocytes, macrophages, dendritic cells, synovial fibroblasts, and endothelial cells in response to stimulation with microbial antigens, immune complexes, and other factors (Hsu, H., Viney, J. The tale of TL1A in inflammation. Mucosal Immunol 4, 368-370 (2011). doi.org / 10.1038 / mi.2011.20).

[0006] The major receptor for TL1A is DR3 (death receptor 3), also known as TNFRSF25 (TNF receptor superfamily member 25).

[0007] The DR3 receptor is mainly expressed on activated lymphocytes such as NK cells and T cells (Aiba Y, Nakamura M. The role of TL1A and DR3 in autoimmune and inflammatory diseases. Mediators Inflamm. 2013; 2013: 258164. doi: 10.1155 / 2013 / 258164. Epub 2013 Dec 21. PMID: 24453414; PMCID: PMC3880748).

[0008] The TL1A-DR3 interaction can induce both the development of inflammatory responses and apoptosis, processes associated with the regulation of innate and adaptive immunity. The DR3 receptor contains a so-called "death domain" (DD) in its cytoplasmic domain. After binding to TL1A, activated DR3 interacts with the adaptor protein TRADD in the cytoplasm via the DD; this then forms a complex with other proteins, such as TRAF2 and RIP1. This receptor-associated signaling complex (also known as Complex I) can activate MAPK signaling pathways, such as ERK, JNK, and p38, as well as NFkB and PI3K pathways. Activation of these signaling pathways promotes increased proliferation and inflammatory responses through increased expression of anti-apoptotic proteins, cytokines, and chemokines. Activation of NFkB leads to the induction of cellular inhibitor of apoptosis proteins 1 / 2 (cIAP1 / 2). Blocking Complex I-mediated activation of anti-apoptotic genes or improper complex formation and / or function leads to DR3-mediated cytotoxicity. Blocking intracellular protein synthesis by the antibiotic actinomycin does not lead to NFkB activation, which in turn leads to the shutdown of apoptosis inhibitor proteins, the dissociation of RIPK1 from complex I, and the interaction of RIPK1 with the FADD adaptor protein and caspase 8. The initiator caspase 8 then activates a cascade of effector caspases 3 and 7, leading to apoptosis. Another variant of cell death development involves the proteins FADD, RIP3, RIP1, and the effector protein MLKL to form an intracellular complex called the necrosome; in this case, the activity of caspase 8 is blocked. It ultimately triggers necroptosis, a form of programmed cell death that is accompanied by a strong immune response.It has been shown that TL1A-DR3 interaction in lymphocytes primarily activates proinflammatory pathways rather than apoptosis (Bittner S, Ehrenschwender M. Multifaceted death receptor 3 signaling-promoting survival and triggering death. FEBS Lett. 2017 Sep;591(17):2543-2555. doi:10.1002 / 1873-3468.12747. Epub 2017 Jul 20. PMID:28686297; L. Wen, L. Zhuang, X. Luo, and P. Wei, “TL1A-induced NF-κB activation and c-IAP2 production prevent DR3-mediated apoptosis in TF-1 cells,” The Journal of Biological Chemistry, Vol. 278, No. 40, pp. 39251-39258, 2003; Bamias G, Jia LG, Cominelli F. Thetumor necrosis factor-like cytokine 1A / death receptor 3 cytokine systeminintestinal inflammation. Curr Opin Gastroenterol. 2013 Nov;29(6):597-602.doi:10.1097 / MOG.0b013e328365d3a2.PMID:24100723).

[0009] TL1A is the only known ligand for DR3, but it also binds to the soluble decoy receptor DcR3, also known as TNFRSF6B (TNF receptor superfamily member 6B). This interaction does not lead to further biological effects but rather serves as a means of maintaining TL1A concentrations in the blood at desired levels. DcR3, in turn, is a secreted receptor that can bind and neutralize three TNF family ligands: FasL, LIGHT, and TL1A.Studies comparing lymphocytes in the lamina propria of patients with chronic enteritis with those from a control group of healthy volunteers have shown that DcR3 is elevated at sites of active enteritis and in the blood of patients; this may be a secondary compensatory mechanism in response to increased expression of proinflammatory TL1A (Xu WD, Li R, Huang AF. Role of TL1A in Inflammatory Autoimmune Diseases: A Comprehensive Review. Front Immunol. 2022 Jul 14;13:891328.doi:10.3389 / fimmu.2022.891328; Bamias G, Jia LG, Cominelli F. The tumor necrosis factor-like cytokine 1A / death receptor 3 cytokine system in intestinal inflammation. Curr Opin Gastroenterol. 2013 Nov;29(6):597-602.doi:10.1097 / MOG.0b013e328365d3a2.PMID:24100723; Zhan C, Patskovsky Y, Yan Q, Li Z, Ramagopal U, Cheng H, Brenowitz M, Hui X, Nathenson SG, Almo SC.Decoystrategies: the structure of TL1A:DcR3 complex.Structure.2011 Feb 9;19(2):162-71.doi:10.1016 / j.str.2010.12.004.PMID:21300286;Spyros I.Siakavellas,Giorgos Bamias,Tumor Necrosis Factor-like Cytokine TL1A and Its Receptors DR3 and DcR3: Important New Factors in Mucosal Homeostasis and Inflammation, Inflammatory Bowel Diseases, Volume 21, Issue 10, October 1, 2015, Pages 2441–2452, https: / / doi.org / 10.1097 / MIB.00000000000000492).

[0010] In intestinal homeostasis, the TL1A-DR3 signaling pathway performs a protective function as part of both innate and adaptive immunity. Disruption of the intestinal epithelial mucosal barrier function leads to contamination of adjacent tissues with microorganisms, whose structural components stimulate antigen-presenting cells (APCs) via Toll-like receptors or Fcγ receptors, thereby increasing TL1A expression and leading to CD4 + and CD8 +TL1A also co-stimulates NK cells and innate lymphoid cells (ILCs). The interaction between TL1A and Tregs constitutively expressing DR3 leads to their proliferation but simultaneously inhibits their suppressive function (Valatas V, Kolios G, Bamias G. TL1A (TNFSF15) and DR3 (TNFRSF25): A Co-stimulatory System of Cytokines With Diverse Functions in Gut Mucosal Immunity. Front Immunol. 2019 Mar 27;10:583. doi:10.3389 / fimmu.2019.00583. PMID:30972074; Spyros I. Siakavellas, Giorgos Bamias, Tumor Necrosis Factor-like Cytokine TL1A and Its Receptors DR3 and DcR3: Important New Factors in Mucosal Homeostasis and Inflammation, Inflammatory BowelDiseases, Volume 21, Issue 10, 1 October 2015, Pages 2441-2452, https: / / doi.org / 10.1097 / MIB.0000000000000492; Wallace KL, Zheng LB, Kanazawa Y, Shih DQ. Immunopathology of inflammatory bowel disease. World J Gastroenterol. 2014 Jan 7; 20(1):6-21.doi:10.3748 / wjg.v20.i1.6; Holmkvist P, RoepstorffK, Uronen-Hansson H, Sandén C, Gudjonsson S, Patschan O, Grip O, MarsalJ, Schmidtchen A, Hornum L, JS, K, Agace WW. A major population of mucosal memory CD4+T cells, coexpressing IL-18Rα and DR3, display innate lymphocyte functionality. MucosalImmunol. 2015 May;8(3):545-58.doi:10.1038 / mi.2014.87.Epub October 1, 2014. PMID: 25269704; PMCID: PMC4424383).

[0011] The TL1A-DR3 signaling pathway is a mediator of several autoimmune diseases, including chronic inflammatory bowel disease (IBD), such as Crohn's disease (CD) and ulcerative colitis (UC). IBD is believed to be caused by a combination of genetic and environmental factors that lead to an excessive and inappropriate immune response against commensal microflora in genetically susceptible individuals. The pathogenesis of chronic intestinal inflammation is due to overexpression of TL1A in APCs in the lamina propria region in response to stimulation by intestinal microflora in patients with Crohn's disease and ulcerative colitis, but not in intestinal tissue from healthy donors. In patients with Crohn's disease, TL1A expression levels correlate with the severity of inflammation, particularly in areas with significant inflammation (Giorgos Bamias, Charles Martin III, Marco Marini. Expression, Localization, and Functional Activity of TL1A, a novel Th1-Polarizing Cytokine in Inflammatory Bowel Disease. J Immunol, 2003 Nov 1, 171(9)4868-4874; doi.org / 10.4049 / jimmunol.171.9.4868).

[0012] Through interaction with DR3, TL1A activates effector T cells, primarily Th1, Th2, Th9, Th17, and all ILC groups. Consequently, immune cells produce cytokines and chemokines, which promote the migration of circulating leukocytes and monocytes from the systemic bloodstream to develop and maintain chronic inflammation in the intestine. TL1A, as a co-stimulatory molecule for IL-12 and IL-18, induces Th1 to produce IFNγ and TNFα; both lead to IFNα-mediated activation of tissue macrophages and additional TNFα expression by the latter. These cytokines trigger apoptosis in intestinal epithelial cells and differentiation of stem cells into myofibroblasts. Activated myofibroblasts are a source of enzymes called matrix metalloproteinases that can degrade extracellular matrix proteins, thereby causing tissue degradation. Th2 cells produce IL-13, which activates NK cells, increases intestinal permeability, and induces apoptosis in intestinal epithelial cells. Th17 cells secrete IL-17A and IL-17F, which are responsible for the migration of neutrophils to the inflammatory site and promote myofibroblast proliferation and collagen deposition, thereby leading to fibrosis (Valatas V, Kolios G, Bamias G. TL1A (TNFSF15) and DR3 (TNFRSF25): A Co-stimulatory System of Cytokines With Diverse Functions in Gut Mucosal Immunity. Front Immunol. 2019 Mar 27;10:583. doi:10.3389 / fimmu.2019.00583. PMID:30972074; Spyros I. Siakavellas, Giorgos Bamias, Tumor Necrosis Factor-like Cytokine TL1A and Its Receptors DR3 and DcR3: Important New Factors in Mucosal Homeostasis and Inflammation, Inflammatory Bowel Diseases, Volume 21, Issue 10, October 1, 2015, Pages 2441–2452, https: / / doi.org / 10.1097 / MIB.00000000000000492; Wallace KL, Zheng LB, Kanazawa Y, Shih DQ. Immunopathology of inflammatory bowel disease. World J Gastroenterol. 2014, January 7;20(1):6–21. doi:10.3748 / wjg.v20.i1.6:Kokkotis G,Bamias G.TL1A as a therapeutic target in inflammatory bowel disease.Expert Rev Clin Immunol.2022 / 6 / 18(6):551-555.doi:10.1080 / 1744666X.2022.2074401.Epub May 12, 2022.PMID:35507314;Baumgart DC,Sandborn WJ.Crohn's disease.Lancet.2012November3:380(9853):1590-605.doi:10.1016 / S0140-6736(12)60026-9.Epub 2012 / 8 / 20.Error:Lancet.2013 / 1 / 19 / 381(9862):204.PMID:22914295;Chen L,Ruan G,Cheng Y,Yi A,Chen D,Wei Y.The role of Th17 cells in inflammatory bowel disease and theresearchprogress.Front Immunol.2023January913:1055914.doi:10.3389 / fimmu.2022.1055914.PMID:36700221;PMCID:PMC9870314;Yamada A,Arakaki R,Saito M,Tsunematsu T,Kudo Y,Ishimaru N.Role of regulatory Tcell in the pathogenesis ofinflammatory bowel disease.World J Gastroenterol.2016Feb 21:22(7):2195-205.doi:10.3748 / wjg.v22.i7.2195.PMID:26900284;PMCID:PMC4734996).

[0013] Therefore, the initial high-level cytokine in the pathogenesis of IBD is TL1A rather than TNFα or any other proinflammatory cytokine. The fact that TL1A co-stimulates T cells and induces the secretion of a wide range of proinflammatory cytokines, including TNFα, but not vice versa may account for the relatively low efficacy of anti-TNFα drugs in IBD therapy (Holmkvist P, Roepstorff K, Uronen-Hansson H et al. A major population of mucosal memory CD4+T cells, coexpressing IL-18Rαand DR3, display innate lymphocyte functionality. Mucosal Immunol. 2015; 8(3):545-558. doi:10.1038 / mi.2014.87; Jin S, Chin J, Seeber S et al. TL1A / TNFSF15 directly induces proinflammatory cytokines, including TNFα, from CD3+CD161+T cells to exacerbate gut inflammation. Mucosal Immunol. 2013;6(5):886-899.doi:10.1038 / mi.2012.124).

[0014] Blockade of the TL1A-DR3 inflammatory signaling pathway may be useful in treating a wide range of inflammatory and / or autoimmune diseases, not limited to Crohn's disease and ulcerative colitis. Thus, this method can be used to treat bronchial asthma, rheumatoid arthritis, systemic lupus erythematosus, psoriasis, ankylosing spondylitis, intestinal fibrosis, liver fibrosis, fibrotic lung diseases, including interstitial lung disease associated with systemic sclerosis (Adam W. Clarke, Lynn Poulton, Doris Shim, David Mabon, Danyal Butt, Matthew Pollard, Vanya Pande, Jean Husten, Jacquelyn Lyons, Chen Tian & Anthony G. Doyle (2018) An anti-TL1A antibody for the treatment of asthma and inflammatory bowel disease, mAbs, 10:4, 664-677, DOI: 10.1080 / 19420862.2018.1440164; Rana Herro, Haruka Miki, Gurupreet S. Sethi, David Mills, Amit Kumar Mehta, Xinh-Xinh Nguyen, Carol Feghali-Bostwick, Marina Miller, David H. Broide, Rachel Soloff, Michael Croft; TL1A PromotesLung Tissue Fibrosis and Airway Remodeling.J Immunol 2020 Nov 1; 205(9):2414-2422. https: / / doi.org / 10.4049 / jimmunol.2000665; Jacob, N., Kumagai, K., Abraham, JP, et al. Direct signaling of TL1A-DR3 on fibroblasts induces intestinal fibrosis in vivo. Sci Rep 10, 18189(2020). https: / / doi.org / 10.1038 / s41598-020-75168-5; Hachulla, E., Agard, C., Allanore, Y., et al. French recommendations for the management of systemic clerosis.OrphanetJRare Dis 16(Suppl 2), 322(2021). https: / / doi.org / 10.1186 / s13023-021-01844-y; Song, YJ., Choi, IA, Meylan, F., et al. Circulating TNF-likeprotein 1A (TL1A) is elevated early in rheumatoid arthritis anddepends onTNF.ArthritisRes Ther22,106(2020).https: / / doi.org / 10.1186 / s13075-020-02198-9;LiL,Fu L,Zhou P,Lu Y,ZhangL,Wang W,Nie J,ZhangD,Liu Y,Wu B,Chen T.Effectsoftumor necrosis factor-like ligand 1A(TL1A)on imiquimod-induced psoriasiformskin inflammation in mice. Arch Dermatol Res. 2020 Sep;312(7):481-490. doi:10.1007 / s00403-019-02030-8. Epub 2020 Jan 18. PMID:31953572).

[0015] Patent documents WO2015073580 and WO2017196663 disclose antibodies against TL1A.

[0016] Related to the above, there is a need to generate antibodies that specifically bind to TL1A.

[0017] Summary of the invention (Detailed description of the invention) The authors of the present invention have developed antibodies that specifically bind to TL1A and have high affinity parameters for binding to TL1A. The antibodies according to the invention are effective inhibitors of activity mediated by the DR3 (TNFRSF25) receptor. The antibodies according to the invention have a high affinity for the FcRn receptor to increase the product circulation time in the patient's blood. In addition, the antibodies according to the invention are stable in human serum and are resistant to high temperatures. The antibodies according to the invention inhibit the release of interferon gamma in the presence of IL-12 and IL-18 and of various proinflammatory cytokines in the presence of IL-12, IL-18 and IL-15. The antibodies according to the invention do not induce direct and cross-linked apoptosis of CHO-tmTL1A cells expressing the membrane-bound form of TL1A and do not lead to the development of cytokine release syndrome.

[0018] Definitions and General Methods Unless defined otherwise herein, all technical and scientific terms used in connection with the present invention shall have the same meanings as commonly understood by one of ordinary skill in the art.

[0019] In addition, unless the context otherwise requires, singular terms shall include plural terms, and plural terms shall include singular terms. Generally, the present classifications and methods of cell culture, molecular biology, immunology, microbiology, genetics, analytical chemistry, synthetic organic chemistry, medical and pharmaceutical chemistry, and hybridization and chemistry of proteins and nucleic acids as described herein are well known to those skilled in the art and are widely used in the art. Enzyme reactions and purification methods are performed according to the manufacturer's instructions, as is common in the art, or as described herein.

[0020] The term "KD" in the present specification refers to an affinity constant (or equilibrium dissociation constant), which is calculated from the ratio of Kd to Ka (ie, Kd / Ka), and is expressed as a molar concentration (M).

[0021] "Binding affinity" generally refers to the intensity of the summation of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, "binding affinity" refers to the intrinsic (characteristic, true) binding affinity of a 1:1 interaction between a pair of members (e.g., an antibody and an antigen). The affinity of a molecule X to its binding partner Y can generally be represented by an equilibrium dissociation constant (KD). Preferred Kd values ​​are about 200nM, 150nM, 100nM, 60nM, 50nM, 40nM, 30nM, 20nM, 10nM, 8nM, 6nM, 4nM, 2nM, 1nM or lower. Affinity can be measured by common methods known in the art, including those described in this specification sheets. Low-affinity antibodies generally slowly bind to antigens and tend to dissociate easily, while high-affinity antibodies generally bind to antigens faster and tend to keep binding for a longer time. A variety of methods are known in the art for measuring binding affinity, any of which can be used for purposes of the present invention.

[0022] The term "Kd", "koff" or "kdis" refers to the dissociation rate constant for a specific interaction between a binding molecule and an antigen. The Koff dissociation rate constant can be measured using bio-layer interferometry: for example, using Octet TM systems; and by surface plasmon resonance: for example, using Biacore TM system.

[0023] The term "Ka," "Kon," or "on-rate" refers to the association rate constant.

[0024] The term "R 2 ” refers to the coefficient of determination.

[0025] The term "response" refers to the antibody-antigen binding signal.

[0026] The term "in vitro" refers to a biological entity, biological process or biological reaction outside the body under artificial conditions. For example, cells grown in vitro are understood to be cells grown in an in vitro environment (e.g., in a test tube, culture vial or microtiter plate).

[0027] The term "ED 50 ” (EC 50 ) (50% effective dose / concentration, half maximal effective concentration) refers to the concentration of an agent that produces 50% of the biological effect (which may include cytotoxicity).

[0028] The terms "anti-TL1A antibody", "antibody to TL1A", "antibody that specifically binds to TL1A" are interchangeable in the context of the present application and refer to an antibody that specifically binds to TL1A.

[0029] Antigen - the protein to which the antibody(ies) under consideration specifically bind.

[0030] Genetic construct (plasmid) - an artificially produced circular DNA molecule that contains the various elements necessary for the expression of a target gene and for replication within an organism.

[0031] Proliferation - the division of cells.

[0032] Reporter Cell Line - A cell line carrying a reporter gene used to assess intracellular signaling.

[0033] Transient production - production of protein from transient producers.

[0034] Transfection - a method for DNA delivery in which foreign DNA molecules are delivered to cells using chemical or physical methods: as part of a liposome complex, as a result of electroporation, etc.

[0035] Chromatography is a method for the separation and analysis of mixtures of substances, as well as the study of their physicochemical properties. It is based on the distribution of substances between two phases: the stationary phase (solid phase or liquid coated on an inert support) and the mobile phase (gas phase or liquid phase, eluent).

[0036] Cytokines - small proteins that regulate the immune response.

[0037] ADCC - Antibody-dependent Cellular Cytotoxicity.

[0038] ADCP - Antibody-dependent cellular phagocytosis.

[0039] C1q-C1q protein complex of the complement system (complement component 1q).

[0040] CD16 - cluster of differentiation 16, also FcγRIIIa, has two isoforms: V158 (or V176) - a high-affinity receptor, and F158 (or F176) - a low-affinity receptor.

[0041] CD32 - Cluster of differentiation 32, also FcγRIIA, has 2 isoforms: H131 - high affinity receptor, R131 - low affinity receptor.

[0042] CD4-cluster of differentiation 4.

[0043] CHO - Chinese Hamster Ovary cell line.

[0044] DcR3 - decoy receptor 3, DR3, and tumor necrosis factor receptor superfamily member 6B, TNFRSF6B).

[0045] DR3 - death receptor 3, DR3, and tumor necrosis factor receptor superfamily member 25, TNFRSF25.

[0046] Fc (Fragment crystallizable) - the constant fragment region of an antibody.

[0047] FcRn - A neonatal receptor for the Fc region of human IgG that protects antibodies from degradation in the lysosomes / endosomes of cells after uptake.

[0048] FcγR - Receptor for the Fc fragment of immunoglobulin G (receptor for the Fc fragment of IgG).

[0049] HRP - horseradish peroxidase.

[0050] LIGHT - a protein, tumor necrosis factor-alpha superfamily member 14, TNFSF14.

[0051] Luc - luciferase.

[0052] NFAT - Nuclear factor of activated T-cells.

[0053] PBMC - peripheral blood mononuclear cells.

[0054] TL1A - TNF-like ligand 1A, also known as tumor necrosis factor alpha superfamily member 15 (TNF superfamily member 15, TNFSF15), a vascular endothelial growth factor inhibitor, also known as VEGI-251. The term TL1A refers to any naturally occurring form of TL1A, whether monomeric or multimeric, including dimers, trimers, etc., which can be derived from any suitable organism. As used herein, the term TL1A refers to TL1A (TNFSF15) from any mammal, such as humans, rats, mice, ferrets, pigs, rabbits, and non-human primates.

[0055] tmTL1A - membrane-bound or transmembrane form of TL1A.

[0056] TNBS - 2,4,6-trinitrobenzenesulfonic acid.

[0057] Tris-HCl is a buffer solution containing tris(hydroxymethyl)aminomethane and hydrochloric acid (HCl).

[0058] HPLC-high performance liquid chromatography.

[0059] VE-vertical electrophoresis.

[0060] IL-12 - interleukin-12.

[0061] IL-13 - Interleukin-13.

[0062] IL-15 - Interleukin-15.

[0063] IL-18 - Interleukin-18.

[0064] IL-5 - interleukin-5.

[0065] IL-6 - interleukin-6.

[0066] ELISA - enzyme-linked immunosorbent assay.

[0067] TNFα - tumor necrosis factor alpha.

[0068] As used in this specification and the claims that follow, unless the context requires otherwise, the words "include" and "comprise" or variations such as "includes", "including", "comprises" or "comprising", will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.

[0069] Antibody The present invention relates to a monoclonal antibody or an antigen-binding fragment thereof that specifically binds to TL1A.

[0070] The term "monoclonal antibody" or "mAb" refers to an antibody that is synthesized and isolated as a discrete clonal population of cells.

[0071] The antibodies of the present invention are recombinant antibodies.

[0072] The term "recombinant antibody" refers to an antibody expressed in a cell or cell line that comprises one or more nucleotide sequences encoding the antibody, wherein the one or more nucleotide sequences are not associated with cells in nature.

[0073] In one aspect, the present invention relates to a monoclonal antibody or an antigen-binding fragment thereof that specifically binds to TL1A, comprising: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 having the amino acid sequence RASQX1X2SX3X4X5LX6, wherein X1=S or G; X2=I or V; X3=S or G; X4=T, S or 0; X5=Y or W; X6=N or A; (ii) having the amino acid sequence X7ASX8X9X 10 X 11 CDR2, where X7=G, A or K; X8=S, T or R; X9=L or R; X 10 =Q or A; X 11 = S, T, or A; and (iii) having the amino acid sequence QQX 12 X 13 SX 14 X 15 X 16 X 17 X 18 X 19 CDR3, where X 12 =A, Y or 0; X 13 =N, G or 0; X 14 =Y, F, or S; X 15 =R, S, P or G; X16 =T, I, L or P; X 17 =L, P, T or S; X 18 =T, P, I or 0; X 19 =D, T or 0; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) having the amino acid sequence X 20 YX 21 X 22 CDR1 of S, X 20 =D, S or G; X 21 =Y, A, or D; X 22 =W or M; (ii) having the amino acid sequence X 23 IX 24 X 25 X 26 GX 27 X 28 TX 29 YX 30 X 31 SX 32 KX 33 CDR2, where X 23 =E, T or A; X 24 =N, Q, A, G, I, L, M, S, T, V or R; X 25 =H, T or S; X 26 =S or G; X 27 =N, R or G; X 28 =0 or S; X 29 =D, T, Y, or S; X 30 =N or A; X 31 =P or D; X 32 =L, V or M; X 33 =S or G; and (iii) having the amino acid sequence X 34 X 35 X 36 X37 X 38 X 39 X 40 X 41 X 42 X 43 YX 44 X 45 X 46 X 47 X 48 CDR3, where X 34 =G, S or 0; X 35 =G, R or 0; X 36 =F, G or 0; X 37 =S, T, R or 0; X 38 =G, S or L; X 39 =S or W; X 40 =P, V, W or T; X 41 =N, P, G or D; X 42 =Y, E, S, or F; X 43 =Y, I, or D; X 44 =A, E, R or 0; X 45 =M, Y, D or 0; X 46 =D or 0; X 47 =V, Y, E or 0; X 48 =Y or 0.

[0074] In some embodiments of the invention, the monoclonal antibody or antigen-binding fragment thereof comprises: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 having the amino acid sequence of SEQ ID NO: 1; (ii) a CDR2 having the amino acid sequence of SEQ ID NO: 6; and (iii) a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 having the amino acid sequence of SEQ ID NO: 16; (ii) a CDR2 having the amino acid sequence EIX1HSGNTDYNPSLKS, wherein X1 = N, Q, A, G, I, L, M, S, T, or V; and (iii) CDR3 having the amino acid sequence of SEQ ID NO: 33.

[0075] In one embodiment of the invention, the antibody according to the invention is an isolated antibody.

[0076] The term "isolated," as used to describe various antibodies according to this specification, refers to an antibody that has been identified and separated and / or regenerated from a cell or cell culture in which the antibody is expressed. Impurities (contaminant components) from the natural environment are materials that generally interfere with the diagnostic or therapeutic use of the polypeptide and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. An isolated polypeptide is generally prepared by at least one purification step.

[0077] As used in this specification, the term "antibody" or "immunoglobulin" (Ig) includes complete antibodies. The term "antibody" refers to a glycoprotein comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain comprises a heavy chain variable region (abbreviated as VH in this specification) and a heavy chain constant region. Each light chain consists of a light chain variable region (abbreviated as VL in this specification) and a light chain constant region. The light chain constant domain can be CK (κ light chain constant domain) or CL (λ light chain constant domain). Preferably, the light chain is a κ (κ) light chain, and the light chain constant domain is preferably CK.

[0078] The antibodies according to the present invention may be of any class (e.g., IgA, IgD, IgE, IgG and IgM, preferably IgG) or subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2, preferably IgG1).

[0079] The VL and VH regions can be further subdivided into hypervariable regions called complementarity determining regions (CDRs), which are located between more conserved regions called framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs, arranged from N-terminus to C-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains form the binding domain that interacts with the antigen.

[0080] The constant regions of the antibodies may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (eg, effector cells) and the first component (CIq) of the classical complement system.

[0081] As used in this specification, the term "antigen-binding portion" or "antigen-binding fragment" of an antibody refers to one or more antibody fragments that retain the ability to specifically bind to an antigen. It is shown that the antigen-binding function of an antibody can be performed by a fragment of a full-length antibody. Examples of binding fragments included in the term "antigen-binding portion" of an antibody include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bond at the hinge region; (iii) an Fd fragment consisting of the VH and CH1 domains; (iv) an Fv fragment consisting of the VL and VH domains of a single arm of an antibody; (v) a dAb fragment (Ward et al., (1989) Nature 341: 544-546), which consists of the VH / VHH domains. In addition, the two regions VL and VH of the Fv-fragment are encoded by different genes, which can be connected using recombinant methods using synthetic linkers that enable them to be accepted as a single protein chain, wherein the VL and VH regions pair to form monovalent molecules (called single-chain Fv (scFv); see, for example, Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). It is assumed that such single-chain molecules are also included in the term "antigen-binding portion" of an antibody. Such antibody fragments are produced using conventional techniques known to those skilled in the art, and these fragments are screened in the same manner as intact antibodies.

[0082] As used in this application, "Kabat numbering scheme" or "numbering according to Kabat" refers to a system for numbering amino acid residues that are more variable (i.e., hypervariable) than other amino acid residues in the variable regions of the heavy and light chains of antibodies (Kabat et al. Ann. NY Acad. Sci., 190:382-93 (1971); Kabat et al. Sequences of Proteins of Immunological Interest, 5th ed., US Department of Health and Human Services, NIH Publication No. 91-3242 (1991)).

[0083] An antibody of the present invention that "specifically binds" a target antigen is one that binds the antigen with sufficient affinity such that the antibody can be used as a diagnostic and / or therapeutic agent targeting proteins or cells or tissues expressing the antigen.

[0084] The term "specifically binds" to a particular polypeptide or an epitope on a particular target polypeptide can be described by way of example by molecules that have a Kd for the target of at least about 200 nM, or at least about 150 nM, or at least about 100 nM, or at least about 60 nM, or at least about 50 nM, or at least about 40 nM, or at least about 30 nM, or at least about 20 nM, or at least about 10 nM, or at least about 8 nM, or at least about 6 nM, or at least about 4 nM, or at least about 2 nM, or at least about 1 nM, or at least about 100 pM or less.

[0085] In one embodiment, the term "specifically binds" refers to binding wherein a molecule binds to a particular polypeptide or epitope on a particular polypeptide without substantially binding to any other polypeptide or epitope on a polypeptide.

[0086] In some embodiments of the invention, the monoclonal antibody or antigen-binding fragment thereof comprises a light chain variable domain comprising a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5.

[0087] In some embodiments of the invention, the monoclonal antibody or antigen-binding fragment thereof comprises a light chain variable domain comprising a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0088] In some embodiments of the invention, the monoclonal antibody or antigen-binding fragment thereof comprises a light chain variable domain comprising a CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15.

[0089] In some embodiments of the invention, the monoclonal antibody or antigen-binding fragment thereof comprises a light chain variable domain comprising: (i) a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5; (ii) a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10; and (iii) a CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15.

[0090] In some embodiments of the invention, the monoclonal antibody or antigen-binding fragment thereof comprises: (i) a light chain variable domain, the light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; or (ii) a light chain variable domain, the light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 2, a CDR2 having the amino acid sequence of SEQ ID NO: 7, and a CDR3 having the amino acid sequence of SEQ ID NO: 12; or (iii) a light chain variable domain, the light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 3, a CDR2 having the amino acid sequence of SEQ ID NO: 8, and a CDR3 having the amino acid sequence of SEQ ID NO: 13; or (iv) a light chain variable domain, the light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO: 9, and a CDR3 having the amino acid sequence of SEQ ID NO: 14; or (v) a light chain variable domain, the light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 5, a CDR2 having the amino acid sequence of SEQ ID NO: 10, and A CDR3 having the amino acid sequence of SEQ ID NO: 15.

[0091] In some embodiments of the invention, the monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, or SEQ ID NO:19.

[0092] In some embodiments of the invention, the monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising a CDR2 having an amino acid sequence selected from the group consisting of: SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, or SEQ ID NO: 32.

[0093] In some embodiments of the invention, the monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising a CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, or SEQ ID NO:36.

[0094] In some embodiments of the invention, the monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising: (i) a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, or SEQ ID NO: 19; (ii) a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, or SEQ ID NO: 32; and (iii) a CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, or SEQ ID NO: 36.

[0095] In some embodiments of the invention, the monoclonal antibody or antigen-binding fragment thereof comprises: (i) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 20, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (ii) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 21, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (iii) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 22, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (iv) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 23, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (v) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 24, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (vi) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 25, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (vii) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 26, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (viii) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 27, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (ix) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 28, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (x) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 29, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (xi) a heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 17, a CDR2 having the amino acid sequence of SEQ ID NO: 30, and a CDR3 having the amino acid sequence of SEQ ID NO: 34; or (xii) a heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 18, a CDR2 having the amino acid sequence of SEQ ID NO: 31, and a CDR3 having the amino acid sequence of SEQ ID NO: 35; or (xiii) a heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 19, a CDR2 having the amino acid sequence of SEQ ID NO: 32, and A CDR3 having the amino acid sequence of SEQ ID NO: 36.

[0096] In some embodiments of the invention, the monoclonal antibody or antigen-binding fragment thereof comprises: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5; (ii) a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10; and (iii) a CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, or SEQ ID NO: 19; (ii) a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, or SEQ ID NO: 32; and (iii) a CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, or SEQ ID NO: 36.

[0097] In some embodiments of the invention, the monoclonal antibody or antigen-binding fragment thereof comprises: (i)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 20, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (ii)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 21, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (iii)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 22, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (iv)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 23, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (v)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 24, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (vi)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 25, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (vii)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 26, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (viii)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 27, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (ix)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 28, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (x)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 29, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (xi)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 2, a CDR2 having the amino acid sequence of SEQ ID NO: 7, and a CDR3 having the amino acid sequence of SEQ ID NO: 12; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 17, a CDR2 having the amino acid sequence of SEQ ID NO: 30, and a CDR3 having the amino acid sequence of SEQ ID NO: 34; or (xii)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 3, a CDR2 having the amino acid sequence of SEQ ID NO: 8, and a CDR3 having the amino acid sequence of SEQ ID NO: 13; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 17, a CDR2 having the amino acid sequence of SEQ ID NO: 30, and a CDR3 having the amino acid sequence of SEQ ID NO: 34; or (xiii)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO: 9, and a CDR3 having the amino acid sequence of SEQ ID NO: 14; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 18, a CDR2 having the amino acid sequence of SEQ ID NO: 31, and a CDR3 having the amino acid sequence of SEQ ID NO: 35; or (xiv)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 5, a CDR2 having the amino acid sequence of SEQ ID NO: 10, and a CDR3 having the amino acid sequence of SEQ ID NO: 15; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 19, a CDR2 having the amino acid sequence of SEQ ID NO: 32, and A CDR3 having the amino acid sequence of SEQ ID NO: 36.

[0098] In some embodiments of the invention, the monoclonal antibody or antigen-binding fragment thereof comprises a light chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, or SEQ ID NO:41.

[0099] In some embodiments of the invention, the monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, or SEQ ID NO:55.

[0100] In some embodiments of the invention, the monoclonal antibody or antigen-binding fragment thereof comprises: (a) a light chain variable domain comprising an amino acid sequence selected from the group consisting of: SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, or SEQ ID NO: 41; and (b) a heavy chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, or SEQ ID NO:55.

[0101] In some embodiments of the invention, the monoclonal antibody or antigen-binding fragment thereof comprises: (i)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 42; or (ii) (a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 43; or (iii) (a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 44; or (iv) (a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 45; or (v)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 46; or (vi)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 47; or (vii)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 48; or (viii)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 49; or (ix)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 50; or (x)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 51; or (xi)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 38, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 52; or (xii)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 39, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 53; or (xiii)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 40, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 54; or (xiv)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO:41, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 55.

[0102] In some embodiments of the invention, the monoclonal antibody that specifically binds to TL1A is a full-length IgG antibody.

[0103] In some embodiments of the invention, the monoclonal antibody that specifically binds to TL1A is a full-length IgG antibody, which is a full-length IgG antibody of the human IgG1, IgG2, IgG3, or IgG4 isotype.

[0104] In some embodiments of the present invention, the monoclonal antibody that specifically binds to TL1A is a full-length IgG antibody, which is a full-length IgG antibody of the human IgG1 isotype.

[0105] In some embodiments of the present invention, the monoclonal antibody comprises the mutations M252Y, S254T, T256E in the Fc fragment in the CH2 region according to the EU numbering scheme for amino acids of antibodies (Edelman GM et al., Proc. Natl. Acad. Sci. USA 63 (1969) pp. 78-85; Kabat, EA et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD, (1991).

[0106] In some embodiments of the invention, the monoclonal antibody comprises the mutations L234A and L235A in the CH2 region according to the EU numbering scheme for antibody amino acids.

[0107] In some embodiments of the invention, the monoclonal antibody comprises deletions 446G and 447K in the CH3 region according to the EU numbering scheme for antibody amino acids.

[0108] In some embodiments of the invention, the monoclonal antibody comprises a light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, or SEQ ID NO:60.

[0109] In some embodiments of the invention, the monoclonal antibody comprises a heavy chain comprising an amino acid sequence selected from the group consisting of: SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, or SEQ ID NO:74.

[0110] In some embodiments of the invention, the monoclonal antibody comprises: (i)(a) a light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, or SEQ ID NO: 60, and (b) a heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, or SEQ ID NO:74.

[0111] In some embodiments of the invention, the monoclonal antibody comprises: (i)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 61; or (ii) (a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 62; or (iii) (a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 63; or (iv) (a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 64; or (v)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 65; or (vi)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 66; or (vii)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 67; or (viii)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 68; or (ix)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 69; or (x)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 70; or (xi)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 57, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 71; or (xii)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 58, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 72; or (xiii)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 59, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 73; or (xiv)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 60, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 74.

[0112] In some embodiments of the invention, the monoclonal antibody that specifically binds to TL1A is an antibody selected from the group consisting of: 01-001, 05-001, 05-002, 05-006, 05-008, 05-010, 05-011, 05-013, 05-014, 05-015, 03-001, 03-002, 04-001, or 04-002.

[0113] In some embodiments of the invention, the monoclonal antibody that specifically binds to TL1A is antibody 01-001.

[0114] Antibody 01-001 contains: (a) a light chain comprising the amino acid sequence of SEQ ID NO: 56; and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 61.

[0115] Antibody 01-001 contains: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 37; (b) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 42.

[0116] Antibody 01-001 contains: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) a CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 11, and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 16, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 20, (iii) CDR3 having the amino acid sequence of SEQ ID NO: 33 (Kabat).

[0117] In some embodiments of the invention, the monoclonal antibody that specifically binds TL1A is antibody 05-001.

[0118] Antibody 05-001 contains: (a) a light chain comprising the amino acid sequence of SEQ ID NO: 56; and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 62.

[0119] Antibody 05-001 contains: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 37; (b) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 43.

[0120] Antibody 05-001 contains: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) a CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 11, and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 16, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 21, (iii) CDR3 having the amino acid sequence of SEQ ID NO: 33 (Kabat).

[0121] In some embodiments of the invention, the monoclonal antibody that specifically binds TL1A is antibody 05-002.

[0122] Antibody 05-002 contains: (a) a light chain comprising the amino acid sequence of SEQ ID NO: 56; and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 63.

[0123] Antibody 05-002 contains: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 37; (b) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 44.

[0124] Antibody 05-002 contains: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) a CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 11, and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 16, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 22, (iii) CDR3 having the amino acid sequence of SEQ ID NO: 33 (Kabat).

[0125] In some embodiments of the invention, the monoclonal antibody that specifically binds to TL1A is antibody 05-006.

[0126] Antibody 05-006 contains: (a) a light chain comprising the amino acid sequence of SEQ ID NO: 56; and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 64.

[0127] Antibody 05-006 contains: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 37; (b) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 45.

[0128] Antibody 05-006 contains: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) a CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 11, and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 16, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 23, (iii) CDR3 having the amino acid sequence of SEQ ID NO: 33 (Kabat).

[0129] In some embodiments of the invention, the monoclonal antibody that specifically binds TL1A is antibody 05-008.

[0130] Antibody 05-008 contains: (a) a light chain comprising the amino acid sequence of SEQ ID NO: 56; and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 65.

[0131] Antibody 05-008 contains: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 37; (b) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 46.

[0132] Antibody 05-008 contains: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) a CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 11, and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 16, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 24, (iii) CDR3 having the amino acid sequence of SEQ ID NO: 33 (Kabat).

[0133] In some embodiments of the invention, the monoclonal antibody that specifically binds to TL1A is antibody 05-010.

[0134] Antibody 05-010 contains: (a) a light chain comprising the amino acid sequence of SEQ ID NO: 56; and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 66.

[0135] Antibody 05-010 contains: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 37; (b) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 47.

[0136] Antibody 05-010 contains: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) a CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 11, and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 16, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 25, (iii) CDR3 having the amino acid sequence of SEQ ID NO: 33 (Kabat).

[0137] In some embodiments of the invention, the monoclonal antibody that specifically binds to TL1A is antibody 05-011.

[0138] Antibody 05-011 contains: (a) a light chain comprising the amino acid sequence of SEQ ID NO: 56; and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 67.

[0139] Antibody 05-011 contains: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 37; (b) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 48.

[0140] Antibody 05-011 contains: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) a CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 11, and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 16, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 26, (iii) CDR3 having the amino acid sequence of SEQ ID NO: 33 (Kabat).

[0141] In some embodiments of the invention, the monoclonal antibody that specifically binds to TL1A is antibody 05-013.

[0142] Antibody 05-013 contains: (a) a light chain comprising the amino acid sequence of SEQ ID NO: 56; and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 68.

[0143] Antibody 05-013 contains: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 37; (b) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 49.

[0144] Antibody 05-013 contains: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) a CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 11, and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 16, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 27, (iii) CDR3 having the amino acid sequence of SEQ ID NO: 33 (Kabat).

[0145] In some embodiments of the invention, the monoclonal antibody that specifically binds to TL1A is antibody 05-014.

[0146] Antibody 05-014 contains: (a) a light chain comprising the amino acid sequence of SEQ ID NO: 56; and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 69.

[0147] Antibody 05-014 contains: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 37; (b) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 50.

[0148] Antibody 05-014 contains: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) a CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 11, and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 16, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 28, (iii) CDR3 having the amino acid sequence of SEQ ID NO: 33 (Kabat).

[0149] In some embodiments of the invention, the monoclonal antibody that specifically binds to TL1A is antibody 05-015.

[0150] Antibody 05-015 contains: (a) a light chain comprising the amino acid sequence of SEQ ID NO: 56; and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 70.

[0151] Antibody 05-015 contains: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 37; (b) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 51.

[0152] Antibody 05-015 contains: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) a CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 11, and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 16, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 29, (iii) CDR3 having the amino acid sequence of SEQ ID NO: 33 (Kabat).

[0153] In some embodiments of the invention, the monoclonal antibody that specifically binds TL1A is antibody 03-001.

[0154] Antibody 03-001 contains: (a) a light chain comprising the amino acid sequence of SEQ ID NO: 57; and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 71.

[0155] Antibody 03-001 contains: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 38; (b) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 52.

[0156] Antibody 03-001 contains: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 7, (iii) a CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 12, and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 17, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 30, (iii) CDR3 having the amino acid sequence of SEQ ID NO: 34 (Kabat).

[0157] In some embodiments of the invention, the monoclonal antibody that specifically binds TL1A is antibody 03-002.

[0158] Antibody 03-002 contains: (a) a light chain comprising the amino acid sequence of SEQ ID NO: 58; and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 72.

[0159] Antibody 03-002 contains: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 39; (b) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 53.

[0160] Antibody 03-002 contains: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 3, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 8, (iii) a CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 13, and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 17, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 30, (iii) CDR3 having the amino acid sequence of SEQ ID NO: 34 (Kabat).

[0161] In some embodiments of the invention, the monoclonal antibody that specifically binds TL1A is antibody 04-001.

[0162] Antibody 04-001 contains: (a) a light chain comprising the amino acid sequence of SEQ ID NO: 59; and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 73.

[0163] Antibody 04-001 contains: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 40; (b) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 54.

[0164] Antibody 04-001 contains: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 4, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 9, (iii) a CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 14, and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 18, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 31, (iii) CDR3 having the amino acid sequence of SEQ ID NO: 35 (Kabat).

[0165] In some embodiments of the invention, the monoclonal antibody that specifically binds TL1A is antibody 04-002.

[0166] Antibody 04-002 contains: (a) a light chain comprising the amino acid sequence of SEQ ID NO: 60; and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 74.

[0167] Antibody 04-002 contains: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 41; (b) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 55.

[0168] Antibody 04-002 contains: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 5, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 10, (iii) a CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 15, and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 19, (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 32, (iii) CDR3 having the amino acid sequence of SEQ ID NO: 36 (Kabat).

[0169] The hypervariable regions (LCDR1, 2, 3 and HCDR1, 2, 3) of the variable domains of the light and heavy chains of all the above-mentioned antibodies are provided according to the Kabat nomenclature. It will be appreciated by those skilled in the art that the hypervariable regions (LCDR1, 2, 3 and HCDR1, 2, 3) of the variable domains of the light and heavy chains may also be represented according to other commonly known numbering schemes, for example, IMGT, Chothia or AbM. Therefore, the present invention also includes all of the above-mentioned antibodies characterized by means of the hypervariable regions (LCDR1, 2, 3 and HCDR1, 2, 3) of the variable domains of the light and heavy chains using the IMGT, Chothia or AbM numbering schemes.

[0170] The antibodies according to the invention that specifically bind to TL1A have high binding affinity parameters for TL1A. The antibodies according to the invention are effective inhibitors of activity mediated by the DR3 (TNFRSF25) receptor. The antibodies according to the invention have a high affinity for the FcRn receptor to increase the product circulation time in the patient's blood. In addition, the antibodies according to the invention are stable in human serum and are resistant to high temperatures. The antibodies according to the invention inhibit the release of interferon gamma in the presence of IL-12 and IL-18 and of various proinflammatory cytokines in the presence of IL-12, IL-18 and IL-15. The antibodies according to the invention do not induce direct and cross-linked apoptosis of CHO-tmTL1A cells expressing the membrane-bound form of TL1A and do not lead to the development of cytokine release syndrome.

[0171] Nucleic acid molecules In one aspect, the present invention relates to a nucleic acid encoding any one of the above antibodies or antigen-binding fragments thereof that specifically binds to TL1A.

[0172] In any of the embodiments, the nucleic acid molecule can be isolated.

[0173] The terms "nucleic acid," "nucleic acid sequence," "nucleic acid sequence," "polynucleotide," "oligonucleotide," "polynucleotide sequence," and "nucleotide sequence" are used interchangeably herein to refer to a precise sequence of nucleotides, whether modified or unmodified, defining a segment or region of a nucleic acid, with or without non-natural nucleotides, and being double-stranded DNA or RNA, single-stranded DNA or RNA, or the transcription products of said DNAs, or the reverse transcription products of said RNAs.

[0174] Unless otherwise indicated, the term nucleotide sequence includes its complement. Thus, a nucleic acid having a specific sequence should be understood to include a nucleic acid having its complementary strand having its complementary sequence.

[0175] An "isolated" nucleic acid molecule is one that has been identified and separated from at least one nucleic acid molecule-impurity. An isolated nucleic acid molecule is different from the form or collection in which it is found under naturally occurring conditions. Thus, an isolated nucleic acid molecule is different from the nucleic acid molecule that exists in a cell under naturally occurring conditions.

[0176] In one aspect, the present invention relates to a nucleic acid molecule comprising a nucleotide sequence encoding an amino acid sequence selected from SEQ ID NO: 75 to 112. The nucleic acid molecule may also comprise any combination of the nucleotide sequences.

[0177] As will be appreciated by those skilled in the art, due to the redundancy of the genetic code, a variety of different DNA sequences may encode the amino acid sequence (VH, VL, CDR, etc.) of the light or heavy chain of an antibody or fragment thereof according to the present invention. It is well within the skill of those skilled in the art to generate these alternative DNA sequences that encode one and the same amino acid sequence. Such variant DNA sequences are within the scope of the present invention.

[0178] In some embodiments of the invention, the nucleic acid is DNA.

[0179] The nucleic acid molecules of the present invention can be isolated from any source that produces monoclonal antibodies or antigen-binding fragments thereof that specifically bind TL1A. In certain embodiments of the present invention, the nucleic acid molecules of the present invention can be synthesized by chemical synthesis rather than isolated.

[0180] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain variable domain of antibodies 01-001, 05-001, 05-002, 05-006, 05-008, 05-010, 05-011, 05-013, 05-014, 05-015, and comprises a nucleotide sequence having SEQ ID NO:75.

[0181] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain variable domain of antibody 03-001, and comprises a nucleotide sequence having SEQ ID NO:76.

[0182] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain variable domain of antibody 03-002, and comprises a nucleotide sequence having SEQ ID NO:77.

[0183] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain variable domain of antibody 04-001, and comprises a nucleotide sequence having SEQ ID NO:78.

[0184] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain variable domain of antibody 04-002, and comprises a nucleotide sequence having SEQ ID NO:79.

[0185] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 01-001, and comprises a nucleotide sequence having SEQ ID NO:80.

[0186] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 05-001, and comprises a nucleotide sequence having SEQ ID NO:81.

[0187] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 05-002, and comprises a nucleotide sequence having SEQ ID NO:82.

[0188] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 05-006, and comprises a nucleotide sequence having SEQ ID NO:83.

[0189] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 05-008, and comprises a nucleotide sequence having SEQ ID NO:84.

[0190] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 05-010, and comprises a nucleotide sequence having SEQ ID NO:85.

[0191] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 05-011, and comprises a nucleotide sequence having SEQ ID NO:86.

[0192] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 05-013, and comprises a nucleotide sequence having SEQ ID NO:87.

[0193] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 05-014, and comprises a nucleotide sequence having SEQ ID NO:88.

[0194] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 05-015, and comprises a nucleotide sequence having SEQ ID NO:89.

[0195] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 03-001, and comprises a nucleotide sequence having SEQ ID NO:90.

[0196] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 03-002, and comprises a nucleotide sequence having SEQ ID NO:91.

[0197] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 04-001, and comprises a nucleotide sequence having SEQ ID NO:92.

[0198] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 04-002, and comprises a nucleotide sequence having SEQ ID NO:93.

[0199] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain of antibodies 01-001, 05-001, 05-002, 05-006, 05-008, 05-010, 05-011, 05-013, 05-014, 05-015, and comprises a nucleotide sequence having SEQ ID NO:94.

[0200] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain of antibody 03-001, and comprises a nucleotide sequence having SEQ ID NO:95.

[0201] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain of antibody 03-002, and comprises a nucleotide sequence having SEQ ID NO:96.

[0202] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain of antibody 04-001, and comprises a nucleotide sequence having SEQ ID NO:97.

[0203] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain of antibody 04-002, and comprises a nucleotide sequence having SEQ ID NO:98.

[0204] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 01-001, and comprises a nucleotide sequence having SEQ ID NO:99.

[0205] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 05-001, and comprises a nucleotide sequence having SEQ ID NO:100.

[0206] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 05-002, and comprises a nucleotide sequence having SEQ ID NO:101.

[0207] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 05-006, and comprises a nucleotide sequence having SEQ ID NO:102.

[0208] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 05-008, and comprises a nucleotide sequence having SEQ ID NO:103.

[0209] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 05-010, and comprises a nucleotide sequence having SEQ ID NO:104.

[0210] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 05-011, and comprises a nucleotide sequence having SEQ ID NO:105.

[0211] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 05-013, and comprises a nucleotide sequence having SEQ ID NO:106.

[0212] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 05-014, and comprises a nucleotide sequence having SEQ ID NO:107.

[0213] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 05-015, and comprises a nucleotide sequence having SEQ ID NO:108.

[0214] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 03-001, and comprises a nucleotide sequence having SEQ ID NO:109.

[0215] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 03-002, and comprises a nucleotide sequence having SEQ ID NO:110.

[0216] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 04-001, and comprises a nucleotide sequence having SEQ ID NO:111.

[0217] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 04-002, and comprises a nucleotide sequence having SEQ ID NO:112.

[0218] The nucleic acid molecules can be used to express recombinant monoclonal antibodies or antigen-binding fragments thereof that specifically bind to TL1A.

[0219] carrier In one aspect, the present invention relates to an expression vector comprising any of the above-described nucleic acid molecules encoding the corresponding amino acid sequence of an antibody that specifically binds to TL1A or a portion thereof (e.g., a heavy chain and / or light chain binding domain sequence). The present invention relates to a vector suitable for expressing any of the nucleotide sequences described herein.

[0220] The term "vector" as used herein refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked.

[0221] As used in this specification, the term "expression" is defined as the transcription and / or translation of a specific nucleotide sequence.

[0222] In some embodiments of the invention, the vector is a plasmid, a circular double-stranded DNA fragment into which additional DNA segments can be inserted.

[0223] In some embodiments of the invention, the vector is a viral (expression) vector, wherein additional DNA segments can be inserted into the viral genome.

[0224] In some embodiments of the present invention, carrier can be introduced into host cell wherein autonomous replication (for example, bacterial vector and the episomal vector of the bacterial site with replication origin).In further embodiments of the present invention, carrier (for example non-episomal vector) can be integrated in the genome of host cell when introducing into host cell, and thereby replicates together with host genome.In addition, some carrier can instruct the expression of the gene that is operably connected with them.This carrier is referred to as " recombinant expression vector " (or abbreviated as " expression vector ") in this article.

[0225] In some embodiments of the present invention, expression vectors include plasmids, retroviruses, adenoviruses, adeno-associated viruses (AAV), plant viruses such as cauliflower mosaic virus, tobacco mosaic virus, cosmids, YACs, and the like. DNA molecules can be inserted into vectors so that the transcriptional and translational control sequences within the vectors perform their intended functions of regulating the transcription and translation of the DNA. Expression vectors and expression control sequences can be selected to be compatible with the host cell used for expression.

[0226] In one embodiment of the present invention, DNA molecules encoding part or all of the heavy and light chain sequences can be inserted into different vectors.

[0227] In one embodiment, any combination of the above-mentioned DNA molecules are introduced into the same expression vector.

[0228] In one embodiment of the invention, the DNA molecule can be introduced into the expression vector by standard methods (eg, ligation of complementary restriction sites on the gene segments of the antibody and vector, or blunt end ligation if no restriction sites are present).

[0229] In some embodiments of the present invention, a suitable vector is a vector comprising restriction sites so that any VH or VL sequence can be easily inserted and expressed, as described above. The recombinant expression vector can also encode a signal peptide that promotes secretion of the antibody chain from the host cell. The antibody chain sequence can be cloned into the vector so that the signal peptide is connected to the N-terminus of the immunoglobulin chain in frame. The signal peptide can be an immunoglobulin signal peptide or a heterologous signal peptide (i.e., a signal peptide from a non-immunoglobulin protein).

[0230] In some embodiments of the present invention, the vector may include an expression control sequence. As used in this specification, the term "expression control sequence" refers to a polynucleotide sequence necessary for achieving expression and processing of the coding sequence into which they are inserted. It will be understood by those skilled in the art that the design of the expression vector (including the selection of expression control sequences) may depend on factors such as the choice of the host cell type to be transformed, the desired antibody expression level, etc. Expression control sequences include appropriate transcription start and stop sequences, promoter sequences, enhancer sequences, and / or insulator sequences; efficient RNA processing signals, such as splice sites and polyadenylation signals; sequences that stabilize cytoplasmic mRNA; sequences that enhance translation efficiency (e.g., Kozak consensus sequences); sequences that enhance protein stability; and sequences that enhance protein secretion, when desired. The characteristics of such expression control sequences vary depending on the host organism; in prokaryotes, such expression control sequences generally include promoters, ribosome binding sites, and transcription termination sequences; in eukaryotes, such expression control sequences generally include promoters and transcription termination sequences. Preferred expression control sequences for expression host cells in mammals include viral elements that ensure high levels of protein expression in mammalian cells, such as promoters and / or enhancers derived from retroviral LTRs (long terminal repeats), cytomegalovirus (CMV) (e.g., CMV enhancer / promoter), simian virus 40 (SV40) (e.g., SV40 promoter / enhancer), adenovirus (e.g., major late promoter adenovirus (AdMLP)), polyomavirus, and strong mammalian promoters such as TTR promoter, natural immunoglobulin promoter, or actin promoter. Expression control sequences include at least all components whose presence is important for expression and processing.

[0231] In some embodiments of the invention, in addition to the antibody chain genes and expression control sequences, the recombinant expression vectors of the invention may carry additional sequences, such as sequences that regulate replication of the vector in host cells (e.g., origins of replication) and selectable marker genes. The selectable marker gene facilitates selection of host cells into which the vector has been introduced.

[0232] host cells In one aspect, the present invention relates to a method for producing a host cell to produce any of the above antibodies or antigen-binding fragments thereof that specifically bind to TL1A, comprising transforming the cell with the above vector.

[0233] In one aspect, the present invention relates to a host cell for producing any of the above-mentioned antibodies or antigen-binding fragments thereof that specifically bind to TL1A, comprising any of the above-mentioned nucleic acids.

[0234] The term "host cell" as used herein refers to a cell into which a recombinant expression vector has been introduced. The present invention relates to a host cell, which may comprise, for example, a vector according to the present invention as described above. The present invention further relates to a host cell comprising, for example, a nucleotide sequence encoding a heavy chain or an antigen-binding portion thereof, a nucleotide sequence encoding a light chain or an antigen-binding portion thereof, or both. It should be understood that "host cell" refers not only to a specific subject cell, but also to the progeny of such a cell. Because modifications may occur in subsequent generations due to mutations or environmental influences, such progeny may actually be different from the parent cell; however, such cells are still included within the scope of the term "host cell" as used herein.

[0235] Nucleic acid molecules encoding monoclonal antibodies or antigen-binding fragments thereof that specifically bind to TL1A, and vectors containing these nucleic acid molecules, according to the present invention, can be used to transfect mammalian cells, plant cells, bacterial cells, or yeast cells. Transfection can be performed by any known method for introducing polynucleotides into host cells. Methods for introducing heterologous polynucleotides into mammalian cells are well known in the art and include dextran-mediated transfection, cationic polymer-nucleic acid complex transfection, transfection by electroporation, calcium phosphate precipitation, 1,5-dimethyl-1,5-diazoundecamethylene polybromide (polybrene)-mediated transfection, protoplast fusion, encapsulation of the polynucleotide in liposomes, and direct microinjection of DNA into the nucleus. In addition, nucleic acid molecules can be introduced into mammalian cells via viral (expression) vectors.

[0236] Mammalian cell lines used as transformation hosts are well known in the art and include multiple immortalized cell lines. These include, for example, Chinese hamster ovary (CHO) cells, NSO cells, SP2 cells, HEK-293T cells, FreeStyle 293 cells (Invitrogen), NIH-3T3 cells, HeLa cells, baby hamster kidney (BHK) cells, African green monkey kidney cells (COS), human hepatocellular carcinoma cells (for example, Hep G2), A549, SK-HEP1, HUH7, Hep-RG cells and many other cell lines. Cell line is selected by determining which cell line has high expression levels and providing the necessary characteristics of produced protein. Other operable cell lines are insect cell lines, for example, Sf9 or Sf21 cells. When a recombinant expression vector encoding a monoclonal antibody or antigen-binding fragment thereof that specifically binds to TL1A is introduced into a mammalian host cell, the antibody or fragment thereof is produced by culturing the host cell for a period of time sufficient to allow expression of the antibody or fragment thereof in the host cell, or more preferably, secretion of the antibody or fragment thereof into the culture medium in which the host cell is grown. The monoclonal antibody or antigen-binding fragment thereof that specifically binds to TL1A can be isolated from the culture medium using standard protein purification techniques. Plant host cells include, for example, Nicotiana, Arabidopsis, duckweed, corn, wheat, potato, and the like. Bacterial host cells include, for example, genera such as Escherichia and Streptomyces. Yeast host cells include Schizosaccharomyces pombe, Saccharomyces cerevisiae, and Pichia pastoris.

[0237] In addition, various known techniques can be used to enhance the production of monoclonal antibodies or antigen-binding fragments thereof that specifically bind to TL1A from production cell lines. For example, the glutamine synthetase gene expression system (GS system) is a commonly used method for enhancing expression under certain conditions.

[0238] Monoclonal antibodies or antigen-binding fragments thereof that specifically bind to TL1A from various cell lines will likely have different glycosylation profiles compared to each other. However, the monoclonal antibodies or antigen-binding fragments thereof that specifically bind to TL1A encoded by the nucleic acid molecules described herein or comprising the amino acid sequences provided herein are part of the present invention regardless of the glycosylation of the binding molecules and generally regardless of the presence or absence of post-translational modifications.

[0239] The above-mentioned host cells do not involve host cells produced using human embryos.

[0240] The above-mentioned host cells do not relate to host cells produced by modifying the genetic integrity of human germline cells.

[0241] Methods for producing antibodies In one aspect, the present invention relates to a method for producing an antibody or antigen-binding fragment thereof that specifically binds to TL1A, comprising culturing the above-described host cell in a growth medium under conditions sufficient to produce the antibody or fragment thereof, followed by isolating and purifying the resulting antibody or fragment thereof.

[0242] Pharmaceutical composition Another aspect of the present invention is a pharmaceutical composition comprising as an active ingredient (or as the only active ingredient) a monoclonal antibody or an antigen-binding fragment thereof according to the present invention that specifically binds to TL1A.

[0243] In one aspect, the invention relates to a pharmaceutical composition comprising any of the above-described antibodies or antigen-binding fragments thereof in combination with one or more pharmaceutically acceptable excipients.

[0244] In one aspect, the present invention relates to a pharmaceutical composition for treating a TL1A-mediated disease or condition, comprising any of the above-described antibodies or antigen-binding fragments thereof in combination with one or more pharmaceutically acceptable excipients.

[0245] In one aspect, the present invention relates to a pharmaceutical composition for treating a TL1A-mediated disease or condition, comprising a therapeutically effective amount of any of the above-described antibodies or antigen-binding fragments thereof in combination with one or more pharmaceutically acceptable excipients.

[0246] "Pharmaceutical composition" means a composition comprising an antibody according to the present invention and at least one component selected from pharmaceutically acceptable and pharmacologically compatible fillers, solvents, diluents, carriers, adjuvants, distributing and sensing agents, and delivery agents.

[0247] The term "pharmaceutically acceptable" refers to one or more compatible liquid or solid components that are suitable for administration in mammals, preferably humans.

[0248] The term "excipient" is used herein to describe any ingredient other than the antibodies according to the invention. These are substances of inorganic or organic nature used in pharmaceutical production / manufacturing to impart the necessary physicochemical properties to the pharmaceutical product.

[0249] In some embodiments, the composition is intended to improve the condition or treat a disease or disorder that may be mediated by TL1A.

[0250] The term "disease or condition mediated by TL1A" refers to any disease or condition that is directly or indirectly associated with TL1A, including the onset, progression, recurrence, or chronification of the disease or condition.

[0251] "Treat," "treatment," and "therapy" refer to a method of alleviating or eliminating a biological disorder and / or at least one of its associated symptoms.

[0252] The term "disorder" means any condition that would benefit from treatment according to the present invention. The definition of the term includes chronic and acute disorders or diseases, including those pathological conditions that predispose a mammal to the disorder in question.

[0253] A "therapeutically effective amount" refers to an amount of a therapeutic agent administered during treatment that will alleviate to some extent one or more of the symptoms of the condition being treated. The therapeutically effective amount can vary depending on factors such as the specific condition being treated, the age, sex, and weight of the patient, and whether the monoclonal antibody or antigen-binding fragment thereof that specifically binds to TL1A is administered as a stand-alone therapy and / or in combination with one or more additional drugs or therapies.

[0254] In one aspect, the subject or patient being treated is a human subject. The subject can be male or female of any age.

[0255] The pharmaceutical composition of the present invention and its preparation method will undoubtedly be obvious to those skilled in the art. The pharmaceutical composition should preferably be manufactured in accordance with GMP (Good Manufacturing Practice) requirements.

[0256] In some embodiments of the pharmaceutical composition, the pharmaceutical composition can include a buffer composition, tonicity agents (osmolytes or osmolytes), stabilizers, and / or solubilizers.

[0257] The pharmaceutical composition according to the present invention is a stable composition.

[0258] A pharmaceutical composition is "stable" if the active agent maintains its physical stability and / or chemical stability and / or biological activity over a specified storage period at a storage temperature, e.g., 2-8°C. Preferably, the active agent maintains both physical and chemical stability, as well as biological activity. The storage period is adjusted based on the results of stability testing under accelerated or natural aging conditions.

[0259] In some embodiments, the pharmaceutical composition is an injectable dosage form.

[0260] In some embodiments, the injectable dosage form is an infusion solution.

[0261] In some embodiments, the injectable dosage form is a solution for subcutaneous administration.

[0262] Injectable preparations can be manufactured in unit dosage form, for example, but not limited to, in ampoules, vials, plastic containers, pre-filled syringes, and autoinjection devices.

[0263] In some embodiments, the pharmaceutical composition is a pharmaceutical composition provided in dry (i.e., powder or granular) form for reconstitution with a suitable solvent (e.g., sterile, pyrogen-free water) prior to administration. Such pharmaceutical preparations can be prepared, for example, by lyophilization, which is known in the art as freeze drying and involves freezing the product followed by removal of the solvent from the frozen material.

[0264] In some embodiments, the pharmaceutical composition is a lyophilizate for preparation of a solution for infusion.

[0265] In some embodiments, the pharmaceutical composition is a lyophilisate for preparation of a solution for subcutaneous administration.

[0266] In some embodiments, the pharmaceutical composition is a concentrate for preparing a solution for infusion.

[0267] In some embodiments, the pharmaceutical composition is a concentrate for preparing a solution for subcutaneous administration.

[0268] In one aspect, the present invention relates to a pharmaceutical composition comprising a monoclonal antibody or antigen-binding fragment thereof according to the present invention that specifically binds to TL1A, and at least one other therapeutically active compound.

[0269] In one aspect, the present invention relates to a pharmaceutical composition for treating a TL1A-mediated disease or condition comprising any of the above-described antibodies or antigen-binding fragments thereof and at least one other therapeutically active compound.

[0270] In one aspect, the present invention relates to a pharmaceutical composition comprising any of the above-mentioned antibodies or antigen-binding fragments thereof and further at least one other therapeutically active compound.

[0271] In one aspect, the present invention relates to a pharmaceutical composition for treating a TL1A-mediated disease or condition, comprising any of the above-mentioned antibodies or antigen-binding fragments thereof and further at least one other therapeutically active compound.

[0272] In one aspect, the present invention relates to a pharmaceutical composition for treating a TL1A-mediated disease or condition, comprising any of the above antibodies or antigen-binding fragments thereof and at least one other therapeutically active compound, wherein the other therapeutically active compound is an antibody, a small molecule, a hormonal therapeutic agent, or any combination thereof.

[0273] In some embodiments of the pharmaceutical composition, the other therapeutically active compound is selected from the group consisting of azathioprine (AZA), 6-mercaptopurine (6-MP), methotrexate, glucocorticoids (GCs), long-acting β2-agonists (LABAs), long-acting anticholinergics (LAACs), hydroxychloroquine, prednisolone, methylprednisolone, belimumab, anifrolumab, acitretin, apremilast, tacrolimus, mycophenolate mofetil, or any combination thereof.

[0274] In some embodiments of the pharmaceutical composition, the TL1A-mediated disease or condition is selected from: Crohn's disease, ulcerative colitis, bronchial asthma, rheumatoid arthritis, systemic lupus erythematosus, psoriasis, ankylosing spondylitis, systemic sclerosis-associated interstitial lung disease (SSc-ILD), intestinal fibrosis, liver fibrosis, or fibrotic lung disease.

[0275] Therapeutic Uses of Monoclonal Antibodies or Antigen-Binding Fragments Thereof That Specifically Bind to TL1A In one aspect, an antibody or antigen-binding fragment thereof that specifically binds to TL1A is used to treat a disease or condition mediated by TL1A.

[0276] In one aspect, the subject or patient being treated is a human subject. The subject can be male or female of any age.

[0277] In one aspect, the present invention relates to a method for treating a TL1A-mediated disease or condition comprising administering any of the above-described antibodies or antigen-binding fragments thereof or the pharmaceutical composition in a therapeutically effective amount to a subject in need of such treatment.

[0278] In one aspect, the invention relates to a method for treating a TL1A-mediated disease or condition comprising administering to a subject in need of such treatment any of the above antibodies or antigen-binding fragments thereof and at least one other therapeutically active compound in therapeutically effective amounts.

[0279] In some embodiments of the method of treatment, the TL1A-mediated disease or condition is selected from: Crohn's disease, ulcerative colitis, bronchial asthma, rheumatoid arthritis, systemic lupus erythematosus, psoriasis, ankylosing spondylitis, systemic sclerosis-associated interstitial lung disease (SSc-ILD), intestinal fibrosis, liver fibrosis, or fibrotic lung disease.

[0280] In some embodiments of the treatment methods, the additional therapeutically active compound is an antibody, a small molecule, a hormonal therapeutic agent, or any combination thereof.

[0281] In one aspect, the present invention relates to the use of the above-mentioned antibody or antigen-binding fragment thereof or the above-mentioned pharmaceutical composition for treating a TL1A-mediated disease or disorder in a subject in need of such treatment.

[0282] In one aspect, the present invention relates to the use of the above-described antibodies or antigen-binding fragments thereof and at least one other therapeutically active compound for treating a TL1A-mediated disease or condition in a subject in need of such treatment.

[0283] In some embodiments of the use, the TL1A-mediated disease or condition is selected from: Crohn's disease, ulcerative colitis, bronchial asthma, rheumatoid arthritis, systemic lupus erythematosus, psoriasis, ankylosing spondylitis, systemic sclerosis-associated interstitial lung disease (SSc-ILD), intestinal fibrosis, liver fibrosis, or fibrotic lung disease.

[0284] In some embodiments of the use, the other therapeutically active compound is an antibody, a small molecule, a hormonal therapeutic agent, or any combination thereof.

[0285] As used herein, a use or method involving an antibody or antigen-binding fragment thereof that specifically binds to TL1A and one or more other therapeutic agents is intended to mean, refer to, and include the following: 1) the simultaneous administration of such combination of an antibody or antigen-binding fragment thereof that specifically binds TL1A and a therapeutic agent to a patient in need of treatment when such components are formulated together as a single dosage form that releases said components to said patient at substantially the same time, 2) when such components are formulated separately from one another into separate dosage forms to be taken by said patient at substantially the same time, such that said components are released to said patient at substantially the same time, such that said combination of an antibody or antigen-binding fragment thereof that specifically binds to TL1A and a therapeutic agent is administered simultaneously to a patient in need of treatment, 3) sequential administration of such combination of an antibody or antigen-binding fragment thereof that specifically binds TL1A and a therapeutic agent to a patient in need of treatment when such components are formulated separately from one another into separate dosage forms to be taken by said patient at consecutive times with significant time intervals between each administration, thereby releasing said components to said patient at substantially different times; and 4) when such components are formulated together into a single dosage form that releases said components in a controlled manner so that they are released to said patient simultaneously, sequentially or co-integrated at the same and / or different times, wherein each component can be administered by the same or different routes, such sequential administration of the combination of said antibody or antigen-binding fragment thereof that specifically binds to TL1A and a therapeutic agent to a patient in need of treatment.

[0286] The antibody or antigen-binding fragment thereof that specifically binds to TL1A can be administered without further therapeutic treatment, ie, as a stand-alone therapy.

[0287] In some embodiments of the methods of treatment or use, an antibody or antigen-binding fragment thereof that specifically binds to TL1A can be administered in combination with another therapeutically active compound selected from the group consisting of azathioprine (AZA), 6-mercaptopurine (6-MP), methotrexate, glucocorticoids (GCs), long-acting β2-agonists (LABAs), long-acting anticholinergics (LAACs), hydroxychloroquine, prednisolone, methylprednisolone, belimumab, aniluomab, acitretin, apremilast, tacrolimus, mycophenolate mofetil, or any combination thereof.

[0288] In some embodiments of the methods of treatment or uses, an antibody or antigen-binding fragment thereof that specifically binds TL1A can be administered in combination with another therapeutically active compound selected from azathioprine, 6-mercaptopurine, methotrexate, or any combination thereof.

[0289] In some embodiments of the treatment methods or uses, an antibody or antigen-binding fragment thereof that specifically binds TL1A can be administered in combination with other therapeutically active compounds selected from the group consisting of glucocorticoids, long-acting β2 agonists, long-acting anticholinergics, or any combination thereof.

[0290] In some embodiments of the method of treatment or use, an antibody or antigen-binding fragment thereof that specifically binds TL1A can be administered in combination with methotrexate.

[0291] In some embodiments of the methods of treatment or uses, an antibody or antigen-binding fragment thereof that specifically binds to TL1A can be administered in combination with another therapeutically active compound selected from hydroxychloroquine, prednisolone, methylprednisolone, azathioprine, belimumab, anirumab, or any combination thereof.

[0292] In some embodiments of the method of treatment or use, an antibody or antigen-binding fragment thereof that specifically binds to TL1A can be administered in combination with another therapeutically active compound selected from the group consisting of acitretin, apremilast, methotrexate, or any combination thereof. In addition, the subject can concomitantly receive topical therapy.

[0293] In some embodiments of the treatment methods or uses, an antibody or antigen-binding fragment thereof that specifically binds TL1A can be administered in combination with another therapeutically active compound selected from methotrexate, tacrolimus, mycophenolate mofetil, azathioprine, or any combination thereof.

[0294] The antibodies or antigen-binding fragments thereof and pharmaceutical compositions specifically binding to TL1A according to the present invention are suitable for parenteral administration in the form of sterile pharmaceutical products, intended for administration into a subject by injection or infusion, by breaching the integrity of the skin or mucous membranes and bypassing the gastrointestinal tract. In particular, parenteral administration is contemplated to include, inter alia, subcutaneous, intraperitoneal, intramuscular, intravenous, intraarterial, intrathecal, intraventricular, intraurethral, ​​intracranial, intrasynovial, and transcutaneous injection or infusion; and renal dialysis infusion techniques.

[0295] In some embodiments of the method of treatment or use, the antibody or antigen-binding fragment or pharmaceutical composition that specifically binds TL1A is administered intravenously.

[0296] In some embodiments, administration is performed intravenously by using infusion, chronic infusion, or sustained continuous infusion.

[0297] In some embodiments of the method of treatment or use, the antibody or antigen-binding fragment or pharmaceutical composition that specifically binds TL1A is administered subcutaneously.

[0298] In some embodiments, subcutaneous administration is performed by using subcutaneous injection.

[0299] In some embodiments of the treatment method or use, a suitable dosage of the monoclonal antibody or antigen-binding fragment thereof that specifically binds to TL1A according to the present invention is in the range of 0.1-200 mg / kg. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 This is a map of an expression vector for transient protein production in eukaryotic cells, which carries the genetic sequence of the human TL1A (TNFSF15) antigen.

[0300] Figure 2 The present invention is a map of an expression vector for transient protein production in eukaryotic cells, which carries the gene sequence of the heavy chain of the antibody against TL1A according to the present invention.

[0301] Figure 3 The present invention is a map of a vector for transient protein production in eukaryotic cells, which carries the gene sequence of the light chain of the antibody against TL1A according to the present invention.

[0302] right Figure 1-3 's comments.

[0303] Figure 4 The present invention is a map of a vector for stable protein production in eukaryotic cells, which carries the gene sequence of the heavy chain of the antibody against TL1A according to the present invention.

[0304] Figure 5 The present invention is a map of a vector for stable protein production in eukaryotic cells, which carries the gene sequence of the light chain of the antibody against TL1A according to the present invention.

[0305] right Figure 4-5 's comments.

[0306] Figure 6 Graphs showing evaluation of the interaction between antibody 01-001 and CHO-tmTL1A cells expressing a membrane-bound form of human TL1A.

[0307] Figure 7 is a graph showing the evaluation of antibody 01-001 for inhibition of interferon gamma (IFNγ) release in response to TL1A on primary cells activated by IL-12 and IL-18.

[0308] Figure 8 is a graph showing the evaluation of antibody 01-001 for inhibition of tumor necrosis factor alpha (TNFα) release in response to TL1A on primary cells activated by IL-12, IL-18, and IL-15.

[0309] Figure 9 is a graph showing the evaluation of antibody 01-001 for inhibition of IL-13 release in response to TL1A on primary cells activated by IL-12, IL-18, and IL-15.

[0310] Figure 10 is a graph showing the evaluation of antibody 01-001 for inhibition of IL-6 release in response to TL1A on primary cells activated by IL-12, IL-18, and IL-15.

[0311] Figure 11 is a graph showing the evaluation of antibody 01-001 for inhibition of IL-17 release in response to TL1A on primary cells activated by IL-12, IL-18, and IL-15.

[0312] Figure 12 is a graph showing the evaluation of antibody 01-001 for inhibition of IL-5 release in response to TL1A on primary cells activated by IL-12, IL-18, and IL-15.

[0313] Figure 13is a graph showing the evaluation of antibody 01-001 for inhibition of GM-CSF release in response to TL1A on primary cells activated by IL-12, IL-18, and IL-15.

[0314] Figure 14 is a graph showing the results comparing the efficacy of antibody 01-001 with a placebo control in a TNBS-induced ulcerative colitis rat model. DETAILED DESCRIPTION

[0315] Example The following examples are provided for a better understanding of the present invention. These examples are for illustration purposes only and should not be construed as limiting the scope of the present invention in any way.

[0316] Materials and general methods General information on the nucleotide sequences of human immunoglobulin light and heavy chains is given in: Kabat, EA et al., Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD (1991). The amino acids of the antibody chains are numbered according to the EU numbering (Edelman, GM et al., Proc. Natl. Acad. Sci. USA 63 (1969) 78-85; Kabat, EA et al., Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, (1991).

[0317] Recombinant DNA techniques use standard methods for manipulating DNA as described in Sambrook, J. et al., Molecular cloning: A laboratory manual; Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1989. Molecular biology reagents are used according to the manufacturers' protocols.

[0318] Gene synthesis prepares the desired gene segment from oligonucleotides prepared by chemical synthesis. Gene segments 300-1400 bp long flanked by single restriction sites are assembled by oligonucleotide annealing and ligation, including PCR amplification, and then cloned via the restriction sites. The DNA sequence of the cloned gene fragment is confirmed by DNA sequencing.

[0319] DNA sequence determination The DNA sequence was determined by Sanger sequencing.

[0320] DNA and protein sequence analysis and sequence data management Unipro's UGENE software suite version 1.29 and SnapGene version 6.1 were used for sequence creation, mapping, analysis, annotation, and illustration.

[0321] Expression Vectors To produce the antibodies described herein, we used variants of genetic constructs designed to express transgenes in eukaryotic cells (e.g., Chinese hamster ovary (CHO) cells) and maintain plasmid copy number in prokaryotic cells (e.g., E. coli). In addition to the transgene encoding the sequence of a given antibody, the vector contains all the elements necessary for protein expression in eukaryotic cells and all the elements necessary for maintaining plasmid copy number in prokaryotic cells.

[0322] Fusion genes comprising the subject antibody chains described below are generated by PCR and / or gene synthesis and assembled by connecting the corresponding nucleic acid segments using known recombinant methods and techniques, for example using unique restriction sites in the corresponding vectors. The resulting nucleic acid sequence is verified by DNA sequencing. For eukaryotic cell transfection, larger quantities of plasmids are prepared by preparing plasmids from transformed E. coli cultures.

[0323] Example 1. Preparation of recombinant TL1A antigen.

[0324] Soluble recombinant human (Homo sapiens), macaque (Macaca mulatta), Javanese macaque (Macaca fascicularis), rat (Rattus norvegicus), and mouse (Mus musculus) TL1A (TNFSF15) proteins were produced by transient transfection in the CHO-K1 suspension cell line. Antigens used for immunization, selection, kinetic, and cell-based assays included secreted forms of the human TL1A (TNFSF15) antigen fused to various peptide tags (https: / / www.uniprot.org / uniprotkb / O95150). For comparison, we also used secreted forms of macaque (https: / / www.uniprot.org / uniprotkb / F6S8F9), Java macaque (https: / / www.uniprot.org / uniprotkb / G7PRK8), rat (https: / / www.uniprot.org / uniprotkb / Q8K3Y7), and mouse (https: / / www.uniprot.org / uniprotkb / Q5UBV8) TNFSF15.

[0325] In order to produce above-mentioned antigen, we use plasmid variant, it is designed for transient expression transgene in eukaryotic cells (for example, Chinese hamster ovary (CHO) cell) and for maintaining copy number in prokaryotic cells (for example, intestinal bacteria).The nucleotide sequence of the gene encoding the above-mentioned antigen is prepared by de novo synthesis.Sequence is used for expressing in Chinese hamster ovary (CHO) cell through codon optimization.Next, use SalI / NotI restriction site or use the molecular cloning technique without ligase that the resulting sequence is cloned in plasmid vector.Check the assembly quality by Sanger sequencing.The resulting genetic construct comprises all essential elements for protein expression in eukaryotic cells, and also has all essential elements that maintain plasmid copy number in prokaryotic cells. Figure 1 An exemplary map of a vector encoding human TL1A as a transgene is shown.

[0326] Antigens were produced in an established cell line (CHO-K1) derived from Chinese hamster ovary cells according to published protocols [Biotechnol Bioeng. 2005 Sep 20;91(6):670-677, Liao M et al., 2004; Biotechnol Lett. 2006 Jun;28(11):843-848; Biotechnol Bioeng. 2003 Nov 5;84(3):332-342]. Cell cultures were reseeded into separate flasks 1 day before transient transfection. After reaching the optimal concentration, the cell cultures were reseeded into the bioreactor for further manipulation. Suspension cultures were performed in flasks using serum-free medium on an orbital shaker. For transient expression, cells were transfected in individual tubes using linear polyethyleneimine (PEI) mixed with plasmid DNA in growth medium B (RPMI). Next, we evaluated transfection efficiency using flow cytometry and techniques known in the art. 48 hours after transient transfection, cell culture supplements were added to the cells and the cells were cultured for 5-6 days. After the incubation period, the culture medium was clarified using a filtration system.

[0327] The resulting antigen is purified in two steps. The first step involves the use of metal chelate affinity chromatography. The second step involves size exclusion chromatography.

[0328] Example 2. Sequences of variable domains of antibodies against TL1A were obtained using human antibody selection in the initial Fab library.

[0329] During the three rounds of selection by phage display, human TL1A (TNFSF15) antigen was immobilized on the surface of specially treated test tubes. For the third round of selection, rat TNFSF15 antigen was immobilized in the same manner. Human TL1A (TNFSF15) antigen was used for the fourth round of selection. The fifth and sixth rounds of selection (if necessary) were performed using both human TL1A antigen and rat / mouse TNFSF15 (for different libraries).

[0330] All of the above antigens were immobilized on the surface of specially treated test tubes. The plastic was then washed with phosphate-buffered saline supplemented with 0.1% Tween-20 and blocked with skim milk or bovine serum albumin in the same phosphate-buffered saline. A primary library of human antibodies was used as the selection library. To this end, it was diluted in phosphate-buffered saline supplemented with skim milk and an additional neutralizing protein with a peptide tag for the TL1A antigen (to screen out nonspecifically bound phage particles). Immunoassay tubes with the immobilized antigens were then incubated with the library.

[0331] Unbound phage particles were removed by multiple washes with a solution of phosphate-buffered saline containing 0.1% Tween 20. Phage particles bound to the antigen were eluted with glycine-HCl buffer (pH 2.2), after which the solution was neutralized with 1M Tris-HCl solution (pH 7.6). Escherichia coli cells were infected with phage, grown in culture medium, and used for the next round of selection. Polyclonal phage enzyme-linked immunosorbent assay (ELISA) was used to enrich the library for the target antigen and to assess the presence of non-specifically bound phage particles.

[0332] For ELISA, high-adsorption plates were immobilized with target human, cynomolgus monkey, Javan macaque, mouse, and rat TL1A (TNFSF15) antigens and with non-target neutral proteins with peptide tags used in the target TL1A antigen. The analysis was performed according to standard ELISA protocols.

[0333] After the fifth and sixth rounds of selection for the target TL1A (TNFSF15) antigen, polyclonal phage ELISA showed sufficient enrichment of the library. For monoclonal sequencing, those libraries that showed a more than 5-fold excess signal for the target TL1A (TNFSF15) antigen compared to a non-homologous control protein were selected. Based on the results of the polyclonal phage ELISA, we selected 9 libraries for which phagemid DNA was isolated from E. coli cultures according to standard manufacturer's procedures (Qiagen). These were then used to transform electrocompetent E. coli cells, followed by cell sorting into single clones, and the unambiguous sequence of the genetic constructs was determined by Sanger sequencing (according to standard procedures). As a result of sequencing, 115 clones were analyzed, containing 21 unique combinations of Fab fragments of the heavy and light chains of the antibodies. Of these, one sequence was selected for further study based on its non-obvious structural features, degree of identity to human sequences, and other criteria.

[0334] Thus the sequence of the variable domain of antibody 01-001 that specifically binds to TL1A (TNFSF15) was generated.

[0335] Example 3. Sequences of variable domains of antibodies against TL1A were obtained using selection from an immune library.

[0336] Recombinant soluble human TL1A antigen with various peptide tags was used to immunize several guanacos (Lamaguanicoe). All animals received alternating antigens with different peptide tags to minimize non-target immune responses to them.

[0337] In some cases, human TL1A antigen in Freund's adjuvant was administered to guanacos once a week over a period of 6 weeks.Before each new administration of antigen, 25 ml of blood was collected for further separation of the peripheral blood mononuclear cell (PBMC) fraction.

[0338] In some cases, guanacos were administered human TL1A antigen in Freund's adjuvant over a period of 16 weeks. Before each new administration of antigen, 25 ml of blood was collected for further separation of the PBMC fraction.

[0339] The obtained sera were analyzed using ELISA according to standard procedures to determine the titer of antibodies against the human TL1A antigen used.Blood samples with antibody titers corresponding to the accepted internal standard were used as starting material for PBMC isolation.

[0340] PBMC fractions were isolated from the blood of immunized animals. Next, total RNA was isolated. The isolated RNA served as a substrate for cDNA synthesis using reverse transcriptase and random hexamer and oligodeoxythymidine oligonucleotides as primers.

[0341] The isolated cDNA was used as the basis for the construction of two Fab-phage libraries containing combinations of sequences of the variable domains of the heavy chains of the immunized guanaco and of human light chains (both kappa and lambda).

[0342] Human TL1A antigen was immobilized on the surface of a specially treated test tube and used for four rounds of selection by phage display. A fifth round was performed with mouse TNFSF15 antigen. Selection was performed using the phage display method described above.

[0343] Sequencing revealed four unique sequences of the Fab fragments of the heavy and light chains of the antibody, all of which were transferred to Koff screening.

[0344] Thus, we obtained the sequences of the variable domains of the following antibodies of the present invention: 03-001, 03-002, 04-001, 04-002.

[0345] Example 4. Obtaining the sequence of the variable domain of an antibody against TL1A that does not contain glycosylation sites in the CDR region.

[0346] To eliminate a potential glycosylation site in CDR2 of the heavy chain variable domain of antibody 01-001, we synthesized a genetic construct encoding the heavy chain variable domain of 01-001 with substitutions in CDR2 (sequences shown in Table 1). We thus generated the sequences of the variable domains of the following antibodies of the present invention: 05-001, 05-002, 05-006, 05-008, 05-010, 05-011, 05-013, 05-014, 05-015. The sequence of the light chain variable domain of antibody 01-001 was used as the light chain variable domain of all the above-mentioned antibodies of the present invention.

[0347] Table 1. Sequences of the CD2 heavy chain variable domains of the antibodies according to the invention (numbering according to Kabat EA et al., 1991). Antibody Heavy chain CDR2 sequence Replacement relative to 01-001 01-001 EINHSGNTDYNPSLKS - 05-001 EIQHSGNTDYNPSLKS N52Q 05-002 EIAHSGNTDYNPSLKS N52A 05-006 EIGHSGNTDYNPSLKS N52G 05-008 EIIHSGNTDYNPSLKS N52I 05-010 EILHSGNTDYNPSLKS N52L 05-011 EIMHSGNTDYNPSLKS N52M 05-013 EISHSGNTDYNPSLKS N52S 05-014 EITHSGNTDYNPSLKS N52T 05-015 EIVHSGNTDYNPSLKS N52V

[0348] Example 5. Construction of a genetic construct encoding a sequence of a full-length antibody against TL1A for production of said sequence in eukaryotic cells.

[0349] The nucleotide sequence of the variable domains of the de novo synthesis antibody. The sequence is codon optimized for expression in Chinese hamster ovary celI. According to standard procedures, the selected sequence of the variable domains of the antibody is cloned into an expression vector containing the sequence of the constant domains of human immunoglobulin IgG1. For this reason, we have produced PCR products of the genes containing the heavy chain and light chain variable domains of the antibody. In the PCR process, the desired overlap (for insertion and the sequence region identical to that of the vector) of 20-30 nucleotides is added to the 5' and 3' ends of the molecule for further molecular cloning by no ligase method.

[0350] The heavy chain variable domain was cloned into a vector containing the sequence encoding the heavy chain constant domain of human immunoglobulin IgG1.

[0351] In some cases, the resulting vector contains all the elements necessary for transient production of a protein in eukaryotic cells, eg, Chinese Hamster Ovary (CHO) cells. Figure 2 This is a schematic diagram of the vector.

[0352] In some cases, the resulting vector contains all the elements necessary for the constitutive production of a protein in eukaryotic cells (eg, Chinese Hamster Ovary (CHO) cells). Figure 4 This is a schematic diagram of the vector.

[0353] The light chain variable domain was cloned into a vector containing the sequence of the light chain constant domain of an IgG1 kappa immunoglobulin.

[0354] In some cases, the resulting vector contains all the elements necessary for transient production of a protein in eukaryotic cells, eg, Chinese Hamster Ovary (CHO) cells. Figure 3 This is a schematic diagram of the vector.

[0355] In some cases, the resulting vector contains all the elements necessary for the constitutive production of a protein in eukaryotic cells (eg, Chinese Hamster Ovary (CHO) cells). Figure 5 This is a schematic diagram of the vector.

[0356] The assembly quality was verified by Sanger sequencing. The resulting genetic construct contained all the elements necessary for protein expression in eukaryotic cells.

[0357] Example 6. Production and purification of antibodies against TL1A.

[0358] In some cases, full-length antibodies were produced in Chinese hamster ovary cell culture (CHO-K1 cell line) using the genetic constructs described above for transient protein production in eukaryotic cells.Example 1 describes the preparation and cultivation of cell cultures.

[0359] In some cases, full-length antibodies are produced in Chinese hamster ovary cell culture (CHO-S cell line) using the genetic constructs described above for constant protein production in eukaryotic cells.

[0360] The full-length antibody was purified in two steps. In the first stage, affinity chromatography was used using a chromatography sorbent with a Protein A ligand. Next, we performed viral inactivation and filtered the solution using a 0.22 μm filter. The second step of chromatography purification was performed using a polybasic anion exchange sorbent.

[0361] The purity of the resulting molecules was assessed using polyacrylamide gel electrophoresis (vertical gel electrophoresis, VEP) using techniques known in the art (Table).

[0362] The purity of the obtained molecules was further assessed by high performance liquid chromatography (HPLC) (Table), as is known in the art.

[0363] Table 2. Monomer content by VEP and HPLC, % Antibody Monomer content (VEP), % Monomer content (HPLC), % 01-001 95.8 87.1 03-001 84.0 98.7 03-002 82.3 99.4 04-001 86.5 97.6 04-002 91.2 96.9 05-001 88.2 83.1 05-002 87.0 87.1 05-006 88.0 83.3 05-008 85.9 80.5 05-010 88.1 76.4 05-011 84.4 76.7 05-013 88.4 83.4 05-014 86.5 80.1 05-015 85.6 79.0 Example 7. Determination of the dissociation constants of the antibodies according to the invention with respect to human and animal TL1A (TNFSF15) by biolayer interferometry.

[0364] The degree of interaction between the resulting antibodies and the TL1A antigen was evaluated by biolayer interferometry on a ForteBio Octet RED 384 (ForteBio) using a dedicated AR2G sensor. The protein was immobilized on the surface of the activated sensor, followed by quenching of unreacted groups, baseline recording, recording of analyte association, and recording of analyte dissociation. The baseline and all other analytical steps were recorded in kinetic buffer (based on Na2HPO4 and KH2PO4, supplemented with NaCl, KCl, 0.1% Tween 20, and 0.1% bovine serum albumin). The resulting data were processed using Octet Data Analysis (Version 9.0) software and a 1:1 interaction model. The results are shown in the table.

[0365] Table 3. Equilibrium dissociation constants (KD) of antibodies and human TL1A based on kinetic testing using ForteBio Thus, all tested antibodies according to the present invention showed a high degree of interaction with human TL1A.

[0366] Table 4. Equilibrium dissociation constants (KD) of antibodies and cynomolgus and Java macaque TL1A (TNFSF15) (with identical sequences) according to kinetic testing using ForteBio. Antibody KD, mol / l kon,1 / (Ms) kdis, 1 / s 01-001 <![CDATA[4.93*10 -12 ]]> <![CDATA[2.16*10 5 ]]> <![CDATA[1.06*10 -6 ]]> 03-001 <![CDATA[1.64*10 -9 ]]> <![CDATA[2.46*10 5 ]]> <![CDATA[4.03*10 -4 ]]> 03-002 <![CDATA[8.63*10 -10 ]]> <![CDATA[2.23*10 5 ]]> <![CDATA[1,93*10 -4 ]]>

[0367] Thus, all of the above-mentioned antibodies according to the invention showed a high degree of interaction with cynomolgus and Javan macaque TL1A (TNFSF15) (sequence identity).

[0368] Table 5. Equilibrium dissociation constants (KD) of antibodies and rat TL1A (TNFSF15) according to kinetic testing using ForteBio Antibody KD, mol / l kon,1 / (Ms) kdis, 1 / s 01-001 <![CDATA[1.44*10 -11 ]]> <![CDATA[2.34*10 5 ]]> <![CDATA[3.38*10 -6 ]]> 03-002 <![CDATA[4.08*10 -9 ]]> <![CDATA[4.63*10 6 ]]> <![CDATA[1.89*10 -2 ]]> Thus, all of the above-mentioned antibodies according to the present invention showed a high degree of interaction with rat TL1A (TNFSF15).

[0369] Example 8. Evaluation of the affinity of the antibodies according to the present invention for human and animal TL1A (TNFSF15) by surface plasmon resonance (SPR).

[0370] The interaction between 01-001 and human and animal TL1A (TNFSF15) antigens was evaluated by surface plasmon resonance (SPR) on a Biacore 8K instrument. The antigen was immobilized on the surface of a carboxymethyl dextran-coated sensor chip (CM5) by amine binding. Human, macaque, Java macaque, green monkey, ferret, pig, rabbit, and rat TL1A (TNFSF15) antigens were diluted in kinetic phosphate buffer (PBS) supplemented with KCl, NaCl, and Tween 20 and passed over the chip surface using a C2H3NaO2-based immobilization buffer (pH 4.0±0.1). Next, the chip surface was activated by introducing an activator mixture, after which the antibody solution was introduced. The immobilization level was determined by changes in surface plasmon resonance. To correct for possible signal distortion, one of the detection channels was used as a reference channel, i.e., the antibody solution was not introduced through the channel at the appropriate stage. The active carboxyl groups were inactivated using a kinetic solution.

[0371] The calculated values ​​of equilibrium dissociation constants (KD) are shown in Table 6.

[0372] Table 6. Equilibrium dissociation constants (KD) of 01-001 with human and animal TL1A (TNFSF15) (including an estimate of amino acid sequence identity between the secreted portions of animal TNFSF15 and human TNFSF15).

[0373] Therefore, antibody 01-001 interacted with human TL1A antigen with high affinity, with a KD value of 7.96×10 -11 M. In addition, 01-001 showed high affinity for TL1A (TNFSF15) in macaques, Javan macaques, green monkeys, ferrets, pigs, rabbits, and rats. Based on these data, rats, Javan macaques, macaques, minipigs, ferrets, green monkeys, and rabbits are relevant species for preclinical studies.

[0374] Example 9. Specificity of binding of antibodies according to the invention to human TL1A (TNFSF15) homologues.

[0375] To evaluate the non-targeted binding of antibody 01-001 to the closest human TL1A homologs, we evaluated the affinity of 01-001 for TRAIL (TNFSF10), FASL (TNFSF6), LIGHT (TNFSF14), EDA-A1, and EDA-A2 proteins, whose extracellular portions are 39%, 35%, 34%, 36%, and 36% identical to the extracellular portion of human TL1A, respectively. Binding affinity was assessed by biolayer interferometry on a ForteBio Octet RED 384 instrument (ForteBio) using a dedicated AR2G sensor according to the above-described procedure. The study included analysis of antigens with various peptide tags. Human TL1A, produced as described above, was used as a positive control.

[0376] The calculated values ​​of equilibrium dissociation constants (KD) are shown in Table 7.

[0377] Table 7. Equilibrium dissociation constants (KD) of 01-001 and human TL1A (TNFSF15) homologs according to kinetic testing using ForteBio Thus, the method showed that antibody 01-001 did not interact with any of the tested proteins of the tumor necrosis factor (TNF) family, but did bind to human TL1A on the same plate used to test homologous proteins. Thus, antibody 01-001 is highly selective for binding to human TL1A and not to its closest homologous proteins of the TNF family.

[0378] Example 10. Determination of the thermal stability of the antibodies according to the present invention.

[0379] The thermal stability of the antibodies was evaluated by heat stress at 50°C for 48 hours in three buffer solutions with different compositions: buffer solution 1 with a pH of 5.0; buffer solution 2 with a pH of 6.0; and buffer solution 3 with a pH of 7.0. Homogeneity was controlled by SEC HPLC, as is known in the art. The results are shown in Tables 7-9.

[0380] Table 7. Studies of intact and isothermal samples of antibody 01-001 in buffer solution 1 at pH 5.0 by HPLC.

[0381] Table 8. Studies of intact and isothermal samples of antibody 01-001 in 20 mM buffer solution 2, pH 6.0, by HPLC. Constant temperature time, h Ratio of monomers, % High molecular weight impurities, % Low molecular weight impurities, % 0 87.2 12.4 0.4 24 86.0 12.0 2.0 36 85.8 12.1 2.1 48 85.5 12.0 2.5

[0382] Table 9. Studies of intact and isothermal samples of antibody 01-001 in buffer solution 3 at pH 7.0 by HPLC. Constant temperature time, h Ratio of monomers, % High molecular weight impurities, % Low molecular weight impurities, % 0 86.3 13.3 0.4 24 84.5 13.6 1.9 36 83.7 14.0 2.3 48 82.0 15.4 2.6

[0383] Thus, during the 48-hour heating period, the change in the main peak area (of monomer 01-001) in the three tested buffer solutions ranged from 2.0 to 4.3%, thereby demonstrating the high thermal stability of antibody 01-001.

[0384] Example 11. Determination of the aggregation temperature of the antibody according to the present invention.

[0385] The aggregation temperature of antibody 01-001 was then determined by dynamic light scattering (DLS) as known in the art.

[0386] The aggregation temperature of the antibody was determined by DLS, after the antibody was incubated at 50°C for 48 hours, and the sample was analyzed by HPLC after incubation. The particle size of the sample was analyzed by DLS. A complete sample of the test antibody in the above buffer solution, which was not incubated at 50°C for 48 hours but incubated at 2-8°C, served as a control. The results of this study are shown in Table 10.

[0387] Table 10. Studies of intact and isothermal samples of antibody 01-001 by DLS using HPLC.

[0388] Note: PD = polydispersity; n / d = not determined.

[0389] The aggregation temperatures of antibody 01-001 determined in the analysis were: 65.92°C for buffer solution 1 (pH 5.0); 66.03°C for buffer solution 2 (pH 6.0); and 63.99°C for buffer solution 3 (pH 7.0).

[0390] Example 12. Determination of the melting temperature of the antibodies according to the present invention.

[0391] The melting temperature of antibody 01-001 was determined by differential scanning fluorimetry (DSF). According to the method, the stability of the sample is proportional to the difference between the temperature of the blank solution (the solution in which the antibody is present) and the sample. The results of the study are shown in Table 11.

[0392] Table 1. Melting temperature of sample 01-001 Note: n / d - not tested.

[0393] The melting temperatures of antibody 01-001 determined in the analysis were: 47.4-60.9°C for buffer solution 1 (pH 5.0); 64.8°C for buffer solution 2 (pH 6.0); and 63.0°C for buffer solution 3 (pH 7.0).

[0394] Example 13. Determination of changes in the kinetic properties of antibodies according to the invention during long-term heating.

[0395] To further evaluate the stability of antibody 01-001, we measured changes in its kinetic properties following the aforementioned 48-hour incubation at 50°C in three different buffer solutions. As described above, the affinity of sample 01-001 for human TL1A was determined using biolayer interferometry on a ForteBio Octet RED 384 instrument and a dedicated AR2G sensor (ForteBio). The results are shown in Table 12.

[0396] Table 2. Equilibrium dissociation constants (KD) of intact and isothermal samples of antibody 01-001 with human TL1A in different buffer solutions Based on the experimental results, antibody 01-001 is highly resistant to long-term heating while maintaining high affinity for human TL1A.

[0397] Example 14. Determination of post-translational modifications of antibodies according to the invention.

[0398] To determine the post-translational modifications of antibody 01-001, we examined the presence and profile of glycosylation, the presence of predicted disulfide bonds, the content and position of trisulfides, thioesters and free cysteines, and their changes after incubation of 01-001 in the three buffer solutions described above at 50°C for 48 hours.

[0399] The analytical experiments determined the post-translational modifications of sample 01-001 by ultra-high performance liquid chromatography with mass spectrometry detection. A post-translational modification library for analysis and processing of all provided samples was constructed using standard samples (N-terminal form of glutamic acid, succinimidyl form of asparagine / glutamine, etc.) and antibody 01-001 in the three aforementioned buffer solutions. Data from the analytical experiments were processed using Protein Metrics software.

[0400] The study confirmed the native molecular weight of antibody 01-001. It showed that in the 01-001 antibody, two standard N-glycosylation sites for IgG1 immunoglobulins (at residue Asn297 in the CH2 heavy chain constant domain) were occupied. No glycosylation of the Fab domain was detected. The results indicate that the theoretical sites for N-glycosylation were not occupied in the CDR2 of the heavy chain variable domain of antibody 01-001.

[0401] Sample 01-001 did not exhibit any oxidized forms of methionine and tryptophan, and no N-terminal glutamate was detected. Analytical experiments confirmed the location of disulfide bonds in 01-001. No improperly cross-linked disulfide bonds, trisulfide compounds, or thioesters were found.

[0402] After incubation of 01-001 at 50°C for 48 hours, no significant changes in post-translational modifications were detected.

[0403] Example 15. Determination of the stability of the antibodies according to the present invention in human serum.

[0404] The stability of 01-001 in normal human serum at 37°C for 14 and 28 days was determined. To this end, we evaluated the ability of antibody 01-001 to bind to human TL1A after incubation under the described conditions using an ELISA according to standard procedures. Briefly, recombinant TL1A was introduced into pretreated wells of a high-absorbance plate, after which human serum samples containing a known amount of antibody 01-001 were added (before incubation at 37°C). After a series of washes, the reaction was developed using a polyclonal antibody against the Fc fragment of human IgG and a conjugate of horseradish peroxidase, and the optical density of the solution was measured using a spectrophotometer at a wavelength of 450 nm. We then determined the amount of 01-001 in the serum samples by comparing the data with a calibration curve that reflects the dependence of the solution optical density on the antibody concentration.

[0405] Studies have shown that human serum retains 70% of the initially introduced molecule 01-001 after 14 days of incubation at 37°C, and 63% after 28 days. The average stability of IgG1 monoclonal antibodies in human serum is generally considered to be 60% functional over a period of one month (28 days) at +37°C. Therefore, the stability of antibody 01-001 is above average.

[0406] Example 16. Interaction between the antibodies according to the invention and the membrane-bound form of human TL1A on the cell surface.

[0407] The 01-001-human TL1A interaction on the surface of eukaryotic cells was evaluated by flow cytometry using a commercially available phycoerythrin (PE)-labeled antibody specific for the human IgG Fc fragment using CHO-tmTL1A cells stably expressing a membrane-bound form of human TL1A. The CHO-K1-S cell line, which does not express a membrane-bound form of human TL1A, was used as a negative control.

[0408] The cell suspension was incubated with serial dilutions of 01-001 on ice in the dark, and then washed from unbound 01-001. Thereafter, the pelleted cells were incubated with a secondary staining antibody against the human IgG Fc fragment on ice in the dark. Next, the cells were washed from the unbound secondary antibody, resuspended in staining buffer, and the fluorescence intensity was measured on a flow cytometer. The dependence of the median fluorescence intensity on the product concentration was evaluated. The results of the study are shown in Figure 6 middle.

[0409] Thus, 01-001 interacts with the membrane-bound form of human TL1A on the surface of eukaryotic cells. The magnitude of the effect is directly dependent on the concentration of 01-001, EC 50 The value was in the nanogram range (163.8 ng / ml).

[0410] Based on the results, CHO-tmTL1A cells stably expressing a membrane-bound form of human TL1A were selected for further in vitro testing to investigate the potential mechanism of action of 01-001 and to evaluate Fc-mediated antibody function.

[0411] Example 17. Inhibition of the interaction between soluble human TL1A and the human DR3 receptor on the surface of cell lines by antibodies according to the invention.

[0412] The effect of the antibody 01-001 on the interaction between soluble TL1A and DR3 on the surface of the TF-1NF-κBLuc cl.4 cell line was determined by flow cytometry using commercially available human biotinylated TL1A. To this end, cell suspensions were incubated with serial dilutions of 01-001 and a solution of biotinylated TL1A. Unbound 01-001 and TL1A were washed away, and the cell pellets were then incubated on ice in the dark with a solution of a phycoerythrin-streptavidin conjugate. Unbound conjugate was washed away, resuspended in staining buffer, and fluorescence intensity was measured on a flow cytometer. The dependence of median fluorescence intensity on product concentration was evaluated. EC 50 It is 1113ng / ml.

[0413] Thus, 01-001 inhibits the interaction between soluble human TL1A (TNFSF15) and human DR3 receptor (TNFRSF25) on the surface of the TF-1NF-κB Luc cl.4 cell line. The magnitude of the effect is directly dependent on the concentration of 01-001, EC 50 The values ​​were in the microgram range (1113 ng / ml).The properties of 01-001 demonstrate that it is effective in preventing the activation of the pro-inflammatory signaling cascade mediated by the TL1A-DR3 interaction.

[0414] Example 18. Comparison of the inhibition of the interaction of soluble human TL1A with the DR3 receptor and the DcR3 decoy receptor by the antibodies according to the present invention.

[0415] The ability of 01-001 to inhibit the interaction of TL1A with DcR3 and DR3 receptors was evaluated using ELISA according to standard procedures. Briefly, soluble proteins of human DR3 (TNFRSF25) and human DcR3 (TNFRSF6B) were immobilized on the surface of plate wells, after which serial dilutions of the 01-001 product were incubated with biotinylated human TL1A. Next, a solution of 01-001 and biotinylated TL1A was added to the immobilized receptors. We used streptavidin-HRP conjugate for detection and measured the optical density (OD) signal on a microplate reader. The percentage of inhibition of TL1A-DR3 and TL1A-DcR3 interactions by 01-001 was determined. The EC values ​​for TL1A-cellular DR3 receptor interactions were quantified. 50 The EC value of TL1A-soluble DcR3 decoy receptor interaction was 1.5 ng / ml. 50 The value is 67.7ng / ml.

[0416] The experimental results showed that antibody 01-001 selectively inhibited the interaction between human TL1A and the target cell DR3 receptor; while its inhibition on the interaction between TL1A and the DcR3 decoy receptor (a natural antagonist of TL1A) was significantly smaller (1 / 45).

[0417] Example 19. Inhibition of caspase-dependent apoptosis mediated by TL1A-DR3 interaction by antibodies according to the invention.

[0418] Under the influence of actinomycin, an inhibitor of the apoptotic protein cIAP2, involved in the NFkB signaling pathway, TL1A binds to the naturally occurring DR3 receptor, leading to caspase activation and induction of apoptosis. The ability of antibodies according to the present invention to inhibit TL1A-mediated apoptosis in the presence of actinomycin was evaluated in a cellular assay using the TF-1 cell line, which expresses the DR3 receptor. To this end, a cell suspension was incubated with serial dilutions of the antibodies according to the present invention and a solution of human TL1A in the presence of actinomycin. A solution of a reagent for detecting caspase-3 / 7 by luminescence intensity was then added, and after further incubation, the luminescence signal was measured on a plate reader. The results of this study are shown in Table 13.

[0419] Table 3. ECs of caspase signaling inhibition in the TF-1 cell line by antibodies according to the invention 50 value. Antibody <![CDATA[EC 50 ,ng / ml]]> Antibody <![CDATA[EC 50 ,ng / ml]]> 01-001 218 05-006 566 03-001 9585 05-008 450 03-002 1914 05-010 334 04-001 943 05-011 508 04-002 421 05-013 >15000 05-001 508 05-014 >15000 05-002 377 05-015 >15000

[0420] Thus, the above antibodies according to the invention inhibit the initiation of apoptosis mediated by the TL1A-DR3 interaction in the presence of actinomycin. The intensity of the effect is directly dependent on the concentration of 01-001, EC 50 Values ​​were in the nanogram range (218-287 ng / ml).

[0421] Example 20. Determination of changes in the inhibitory properties of the antibodies according to the present invention under long-term heating.

[0422] To further evaluate the stability of antibody 01-001, we determined its functional properties for inhibiting TL1A-mediated apoptosis after incubation at 50°C for 48 hours in three different buffer solutions. The inhibitory activity was determined using the caspase-3 / 7 luminescence assay described above. The results of this study are shown in Table 14.

[0423] Table 4. ECs for inhibition of caspase signaling in the TF-1 cell line by intact and isothermal samples of antibody 01-001 in different buffer solutions 50 and EC 90 value Based on the experimental results, antibody 01-001 has remarkable resistance to long-term heating while maintaining its biological properties with respect to inhibition of caspase signaling mediated by TL1A-DR3 interaction.

[0424] Example 21. Inhibition of TL1A-DR3 interaction-mediated activation of the NFkB signaling pathway by antibodies according to the invention.

[0425] To further demonstrate the effects of the antibodies according to the present invention on TL1A-mediated DR3 activation, the inhibitory efficacy and NFkB signaling pathway were evaluated. To this end, cell suspensions of the TF-1 NF-kB Luc cl.4 reporter line, which expresses the DR3 receptor and carries the luciferase gene under the control of an NFkB-responsive element, were incubated with serial dilutions of the antibodies according to the present invention and solutions of human TL1A at the same concentration. Luciferase was detected by measuring the luminescence signal on a microplate reader. The results of this study are shown in Table 15.

[0426] Table 5. ECs of the NFkB signaling pathway of the TF-1 NF-kB Luc cl.4 cell line inhibited by the antibodies according to the present invention 50 value. Antibody <![CDATA[EC 50 ,μg / ml]]> 01-001 0.72 03-001 9.67 03-002 6.36

[0427] Thus, the antibodies of the present invention effectively inhibit the TL1A-DR3 interaction-mediated activation of the NFkB signaling pathway. The effect is directly dependent on the concentration of the product; EC 50 Values ​​are in the nanogram and microgram concentration range.

[0428] Example 22. Inhibition of interferon gamma release in response to TL1A in the presence of IL-12 and IL-18 by antibodies according to the invention.

[0429] IL-12 and IL-18 have been shown to induce TCR (T cell receptor)-independent production of interferon gamma (IFNγ) in CD4+ T lymphocytes, and increased concentrations of soluble TL1A or IL-15 (e.g., in the setting of inflammatory bowel disease) can enhance these effects.

[0430] The ability of 01-001 to inhibit the release of IFNγ in response to TL1A was evaluated using an enzyme-linked immunosorbent assay (ELISA) on IL-12 / IL-18-activated PBMCs isolated from whole blood of healthy volunteers. Briefly, PBMCs were incubated for 20-24 hours with a solution of human cytokines TL1A, IL-12, IL-18 and serial dilutions of 01-001. PBMCs activated by IL-12 and IL-18 (in the absence of TL1A) were used as controls for TL1A-independent release of IFNγ; inactive PBMCs (cell control) were used as negative controls for IFNγ release. After the incubation period, the amount of IFNγ in the supernatant was determined using ELISA, and the optical density signal was measured on a microplate reader. The results are shown in Figure 7 middle.

[0431] Thus, 01-001 inhibits the release of IFNγ in response to TL1A on primary blood cells activated by IL-12 and IL-18. The effect is directly dependent on the concentration of the product 01-001; EC 50 The value was in the nanogram concentration range (496.5 ng / ml).

[0432] Example 23. Inhibition of cytokine release in response to TL1A in the presence of IL-12, IL-18 and IL-15 by antibodies according to the invention.

[0433] It has been shown that the presence of soluble TL1A together with IL-12, IL-18 and IL-15 leads to a markedly increased production of many proinflammatory cytokines such as IFNγ, IL-6, GM-CSF, IL-5, IL-13, TNF-α and IL-22 in CD45RO+CD4+ T-lymphocytes.

[0434] The ability of 01-001 to inhibit the release of cytokines TNF-α, IL-13, IL-6, IL-17, IL-5, and GM-CSF in response to TL1A was evaluated using a multiplex assay on T-lymphocytes isolated from IL-12 / IL-18 / IL-15-activated PBMCs of healthy volunteers. T-lymphocytes activated by IL-12, IL-18, and IL-15 in the absence of TL1A served as controls for TL1A-independent cytokine release, while inactivated T-lymphocytes served as a negative control for cytokine release.

[0435] T-lymphocytes were incubated with a solution of human cytokines TL1A, IL-12, IL-18, IL-15 and serial dilutions of 01-001 for 96 hours. After the incubation time, the amount of cytokines TNF-α, IL-13, IL-6, IL-17, IL-5 and GM-CSF in the supernatant was determined using a multiplex assay and the fluorescence signal was measured. The results are shown in Figure 8-13 middle.

[0436] Thus, antibody 01-001 inhibits the release of proinflammatory cytokines TNF-α, IL-13, IL-6, IL-17, IL-5, GM-CSF from blood cells in response to TL1A in combination with IL-12, IL-18 and IL-15. The effect is directly dependent on the concentration of product 01-001; EC 50 The values ​​were in the nanogram concentration range (12.07-126.6 ng / ml).Based on the results, use of 01-001 will result in reduced severity of symptoms of a TL1A-mediated disease or disorder.

[0437] Example 24. Affinity of the antibodies according to the present invention for Fcγ receptors The affinity of 01-001 for Fcγ receptors (FcγRIIa-131H, FcγRIIa-131R, FcγRIIIa-158V, FcγRIIIa-158F, FcγRIIb, FcγRIIIb) was determined by analyzing changes in surface plasmon resonance (SPR). The affinity of 01-001 for FcγRIa was determined by biolayer interferometry (BLI). The calculated equilibrium dissociation constant (KD) is shown in Table 16.

[0438] Table 16. Equilibrium dissociation constants (KD) of 01-001 for Fcγ receptors *KD value not determined due to low affinity binding Therefore, 01-001 has low affinity for all Fcγ receptors.

[0439] Example 25. Affinity of antibodies according to the invention for FcRn.

[0440] Evaluation of the affinity of a product for the FcRn receptor is important because the time a product circulates in the bloodstream and its half-life in vivo is affected by the extent of binding to the receptor.

[0441] Comparative analysis of the affinity of 01-001 for the FcRn receptor was performed on a ForteBio Octet RED 384 instrument using a dedicated streptavidin SA biosensor (ForteBio) according to the above procedure. For control analysis, we used the replicated antibody PF-06480605 against TL1A according to patent WP2021260577 (Pfizer Inc.) and the replicated antibody adalimumab against TNF-α (tumor necrosis factor α). The calculated values ​​of the equilibrium dissociation constant (KD) are shown in Table 17.

[0442] Table 17. Affinity of antibodies for the FcRn receptor according to the results of kinetic assay using ForteBio. Antibody KD, mol / l kon,1 / (Ms) kdis, 1 / s 01-001 <![CDATA[1.20*10 -9 ]]> <![CDATA[1.27*10 6 ]]> <![CDATA[1.53*10 -3 ]]> PF-06480605 <![CDATA[1.04*10 -8 ]]> <![CDATA[2.14*10 6 ]]> <![CDATA[2.23*10 -2 ]]> Adalimumab <![CDATA[1.17*10 -8 ]]> <![CDATA[1.78*10 6 ]]> <![CDATA[2.09*10 -2 ]]>

[0443] Conclusion: The test antibodies according to the present invention showed high affinity to FcRn over the control antibody.

[0444] Example 26. Affinity of antibodies according to the invention for C1q.

[0445] The affinity of 01-001 for C1q was determined by analyzing changes in surface plasmon resonance (SPR). Due to the low affinity of 01-001 for C1q, the KD value was not determined.

[0446] Example 27. Evaluation of antibody-dependent cellular cytotoxicity (ADCC) induced by the antibodies according to the present invention.

[0447] The ability of 01-001 to induce antibody-dependent cellular cytotoxicity against cells expressing a membrane-bound form of TL1A was evaluated in a reporter cell assay. CHO-tmTL1A cells were used as target cells, and Jurkat-based reporter cell lines carrying a luciferase-encoding gene under the control of an NFAT-responsive element and expressing either a high-affinity or low-affinity CD16 receptor were used as effector cells. Suspensions of the target and effector cell lines were incubated with serial dilutions of the 01-001 product, and the luminescent signal was measured on a microplate reader.

[0448] The results of the experiments showed that 01-001 did not induce antibody-dependent cellular cytotoxicity mediated by both high-affinity and low-affinity CD16 receptors against CHO-tmTL1A cells expressing the membrane-bound form of TL1A.

[0449] Example 28. Evaluation of antibody-dependent cellular phagocytosis (ADCP) induced by the antibodies according to the present invention.

[0450] The ability of 01-001 to induce antibody-dependent cellular phagocytosis of cells expressing a membrane-bound form of TL1A was evaluated in a reporter cell assay. CHO-tmTL1A cells were used as target cells, and a Jurkat-based reporter cell line carrying a luciferase-encoding gene under the control of an NFAT-responsive element and expressing either the high-affinity receptor CD32 or the low-affinity receptor CD32 was used as the effector cell line. Suspensions of the target and effector cell lines were incubated with serial dilutions of 01-001, and the luminescent signal was measured on a microplate reader.

[0451] The experimental results showed that 01-001 did not induce antibody-dependent cellular phagocytosis against CHO-tmTL1A cells expressing the membrane-bound form of TL1A.

[0452] Example 29. Evaluation of complement-dependent cytotoxicity (CDC) induced by antibodies according to the present invention.

[0453] The ability of 01-001 to induce complement-dependent cytotoxicity was evaluated using a cellular assay that determines CDC based on reduced cell viability and the ability to metabolize the resazurin vital dye, which in turn results in a reduced fluorescent signal.

[0454] CHO-tmTL1A expressing a membrane-bound form of TL1A was used as target cells.

[0455] The results of the experiments showed that 01-001 did not induce complement-dependent cytotoxicity against CHO-tmTL1A cells expressing the membrane-bound form of TL1A.

[0456] Example 30. Evaluation of direct and cross-linked apoptosis induction by antibodies according to the invention.

[0457] The ability of 01-001 to induce direct and cross-linked apoptosis against CHO-tmTL1A cells expressing a membrane-bound form of TL1A was determined by flow cytometry using Annexin V to detect apoptotic cells.

[0458] To assess direct apoptosis, serial dilutions of 0.1-0.01 were added to a CHO-tmTL1A cell suspension and incubated for 24 hours. To assess cross-linked apoptosis, an Fc-specific cross-linking antibody was added to the system, followed by incubation with a mixture of PE (phycoerythrin)-labeled Annexin V and 7-aminoactinomycin D at room temperature in the dark. A 1% hydrogen peroxide solution was used as a positive control. The percentage of the Annexin V-positive population was determined on a flow cytometer.

[0459] According to the results, 01-001 did not induce direct and cross-linked apoptosis in CHO-tmTL1A cells expressing the membrane-bound form of TL1A.

[0460] Example 31. Evaluation of changes in cytokine release levels in response to PBMC exposure to antibodies according to the invention.

[0461] Evaluation of cytokine release in response to exposure to 01-001 leads to conclusions about the potential risk of developing cytokine release syndrome and therefore has prognostic value for demonstrating the safety of the present invention.

[0462] The ability of 01-001 to induce the release of 23 cytokines was determined using PBMCs from 10 healthy volunteers: eotaxin, GM-CSF, IFNα2, IFNγ, IL-10, IL-12p40, IL-12p70, IL-13, IL-15, IL-17A, IL1RA, IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-7, MIP-1α, MIP-1β, TNFα, TNFβ, VEGF. HUVEC monolayers were incubated with PBMCs and 01-001 for 24 hours; after the incubation period, the amount of cytokines in the supernatant was determined using a multiplex assay.

[0463] According to the results, the use of 01-001 did not lead to the development of cytokine release syndrome.

[0464] Example 32. Efficacy of antibodies according to the invention in the treatment of inflammatory diseases of the mammalian intestine.

[0465] The efficacy of antibody 01-001 in vivo was determined in a TNBS-induced ulcerative colitis model. Rats received four doses of trinitrobenzenesulfonic acid (TNBS) in 40% ethanol via intrarectal administration on postnatal days 23, 35, 48, and 59. Control animals received an equal volume of ethanol without TNBS. All animals developed focal colitis characterized by colonic ulcers with inflammatory infiltrates and varying degrees of fibrosis. Antibody treatment was initiated a few days after the onset of the colitis challenge (postnatal days 26-57).

[0466] The response to treatment was determined by changes in animal weight, stool consistency, and bleeding, and we also performed histology of large intestine sections to determine the degree of inflammation. A semi-quantitative scoring scale for general intestinal inflammation was used to evaluate the efficacy of antibody 01-001: 1) Severity of inflammation: absent = 0, mild = 1, moderate = 2, severe = 3.

[0467] 2) Depth of inflammation: absent = 0, mucosa only = 1, mucosa and submucosa = 2, transmural = 3.

[0468] 3) Crypth damage: one-third damaged = 1, two-thirds damaged = 2, no crypts but surface epithelium present = 3, no crypts and surface epithelium = 4.

[0469] 4) Proportion of visual fields affected by inflammation: none = 0, 1-25% = 1, 26-50% = 2, 51-75% = 3, 76-100% = 4.

[0470] We evaluated six independent fields, added their scores, and assigned the total to a single animal. The evaluation results are shown in Table 18.

[0471] Table 18. Cumulative scores of intestinal inflammation for individual animals.

[0472] Thus, administration of antibody 01-001 significantly reduced various symptoms of the disease (p<0.002 according to the Mann-Whitney U test, Figure 14 ).

Claims

1. A monoclonal antibody or antigen-binding fragment thereof that specifically binds to TNF-like ligand 1A (TL1A), comprising: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 having the amino acid sequence RASQX1X2SX3X4X5LX6, wherein X1=S or G; X2=I or V; X3=S or G; X4=T, S or 0; X5=Y or W; X6=N or A; (ii) having the amino acid sequence X7ASX8X9X 10 X 11 CDR2, where X7=G, A or K; X8=S, T or R; X9=L or R; X 10 =Q or A; X 11 =S, T or A; and (iii) having the amino acid sequence QQX 12 X 13 SX 14 X 15 X 16 X 17 X 18 X 19 CDR3, X 12 =A, Y or 0; X 13 =N, G or 0; X 14 =Y, F, or S; X 15 =R, S, P or G; X 16 =T, I, L or P; X 17 =L, P, T or S; X 18 =T, P, I or 0; X 19 =D, T or 0; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) having the amino acid sequence X 20 YX 21 X 22 CDR1 of S, X 20 =D, S or G; X 21 =Y, A, or D; X 22 =W or M; (ii) having the amino acid sequence X 23 IX 24 X 25 X 26 GX 27 X 28 TX 29 YX 30 X 31 SX 32 KX 33 CDR2, where X 23 =E, T or A; X 24 = N, Q, A, G, I, L, M, S, T, V or R; X 25 =H, T or S; X 26 =S or G; X 27 =N, R or G; X 28 =0 or S; X 29 =D, T, Y, or S; X 30 =N or A; X 31 =P or D; X 32 =L, V or M; X 33 =S or G; and (iii) having the amino acid sequence X 34 X 35 X 36 X 37 X 38 X 39 X 40 X 41 X 42 X 43 YX 44 X 45 X 46 X 47 X 48 CDR3, X 34 =G, S or 0; X 35 =G, R or 0; X 36 =F, G or 0; X 37 =S, T, R or 0; X 38 =G, S or L; X 39 =S or W; X 40 =P, V, W or T; X 41 =N, P, G or D; X 42 =Y, E, S, or F; X 43 =Y, I, or D; X 44 =A, E, R or 0; X 45 =M, Y, D or 0; X 46 =D or 0; X 47 =V, Y, E or 0; X 48 =Y or 0.

2. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, comprising: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 having the amino acid sequence of SEQ ID NO: 1; (ii) a CDR2 having the amino acid sequence of SEQ ID NO: 6; and (iii) a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 having the amino acid sequence of SEQ ID NO: 16; (ii) a CDR2 having the amino acid sequence EIX1HSGNTDYNPSLKS, wherein X1 = N, Q, A, G, I, L, M, S, T, or V; and (iii) CDR3 having the amino acid sequence of SEQ ID NO:

33.

3. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the light chain variable domain comprises a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO:

5. 4 . The monoclonal antibody or antigen-binding fragment thereof according to claim 1 , wherein the light chain variable domain comprises a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO:

10.

5. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the light chain variable domain comprises a CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO:

15.

6. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the light chain variable domain comprises: (i) a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5; (ii) a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10; and (iii) a CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO:

15.

7. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, comprising: (i) a light chain variable domain, the light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; or (ii) a light chain variable domain, the light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 2, a CDR2 having the amino acid sequence of SEQ ID NO: 7, and a CDR3 having the amino acid sequence of SEQ ID NO: 12; or (iii) a light chain variable domain, the light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 3, a CDR2 having the amino acid sequence of SEQ ID NO: 8, and a CDR3 having the amino acid sequence of SEQ ID NO: 13; or (iv) a light chain variable domain, the light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO: 9, and a CDR3 having the amino acid sequence of SEQ ID NO: 14; or (v) a light chain variable domain, the light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 5, a CDR2 having the amino acid sequence of SEQ ID NO: 10, and A CDR3 having the amino acid sequence of SEQ ID NO:

15.

8. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the heavy chain variable domain comprises a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, or SEQ ID NO:

19.

9. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the heavy chain variable domain comprises a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, or SEQ ID NO:

32.

10. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the heavy chain variable domain comprises a CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, or SEQ ID NO:

36.

11. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the heavy chain variable domain comprises: (i) a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, or SEQ ID NO: 19; (ii) a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, or SEQ ID NO: 32; and (iii) a CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, or SEQ ID NO:

36.

12. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, comprising: (i) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 20, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (ii) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 21, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (iii) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 22, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (iv) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 23, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (v) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 24, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (vi) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 25, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (vii) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 26, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (viii) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 27, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (ix) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 28, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (x) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 29, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (xi) a heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 17, a CDR2 having the amino acid sequence of SEQ ID NO: 30, and a CDR3 having the amino acid sequence of SEQ ID NO: 34; or (xii) a heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 18, a CDR2 having the amino acid sequence of SEQ ID NO: 31, and a CDR3 having the amino acid sequence of SEQ ID NO: 35; or (xiii) a heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 19, a CDR2 having the amino acid sequence of SEQ ID NO: 32, and A CDR3 having the amino acid sequence of SEQ ID NO:

36.

13. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, comprising: (a) a light chain variable domain, the light chain variable domain comprising: (i) a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5; (ii) a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10; and (iii) a CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, or SEQ ID NO: 19; (ii) a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, or SEQ ID NO: 32; and (iii) a CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, or SEQ ID NO:

36.

14. The monoclonal antibody or antigen-binding fragment thereof according to claim 13, comprising: (i)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 20, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (ii)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 21, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (iii)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 22, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (iv)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 23, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (v)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 24, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (vi)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 25, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (vii)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 26, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (viii)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 27, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (ix)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 28, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (x)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 16, a CDR2 having the amino acid sequence of SEQ ID NO: 29, and a CDR3 having the amino acid sequence of SEQ ID NO: 33; or (xi)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 2, a CDR2 having the amino acid sequence of SEQ ID NO: 7, and a CDR3 having the amino acid sequence of SEQ ID NO: 12; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 17, a CDR2 having the amino acid sequence of SEQ ID NO: 30, and a CDR3 having the amino acid sequence of SEQ ID NO: 34; or (xii)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 3, a CDR2 having the amino acid sequence of SEQ ID NO: 8, and a CDR3 having the amino acid sequence of SEQ ID NO: 13; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 17, a CDR2 having the amino acid sequence of SEQ ID NO: 30, and a CDR3 having the amino acid sequence of SEQ ID NO: 34; or (xiii)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO: 9, and a CDR3 having the amino acid sequence of SEQ ID NO: 14; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 18, a CDR2 having the amino acid sequence of SEQ ID NO: 31, and a CDR3 having the amino acid sequence of SEQ ID NO: 35; or (xiv)(a) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 5, a CDR2 having the amino acid sequence of SEQ ID NO: 10, and a CDR3 having the amino acid sequence of SEQ ID NO: 15; and (b) a heavy chain variable domain, the heavy chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 19, a CDR2 having the amino acid sequence of SEQ ID NO: 32, and A CDR3 having the amino acid sequence of SEQ ID NO:

36.

15. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the light chain variable domain comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, or SEQ ID NO:

41.

16. The monoclonal antibody or antigen-binding fragment thereof of claim 1, wherein the heavy chain variable domain comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, or SEQ ID NO:

55.

17. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein: (a) the light chain variable domain comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, or SEQ ID NO: 41; and (b) the heavy chain variable domain comprises an amino acid sequence selected from the group consisting of SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, or SEQ ID NO:

55.

18. The monoclonal antibody or antigen-binding fragment thereof according to claim 17, wherein: (i)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 42; or (ii) (a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 43; or (iii) (a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 44; or (iv) (a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 45; or (v)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 46; or (vi)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 47; or (vii)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 48; or (viii)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 49; or (ix)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 50; or (x)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 51; or (xi)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 38, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 52; or (xii)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 39, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 53; or (xiii)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO:40, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 54; or (xiv)(a) the light chain variable domain comprises the amino acid sequence of SEQ ID NO:41, and (b) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO:

55.

19. The monoclonal antibody of any one of claims 1-18, wherein the antibody that specifically binds to TL1A is a full-length IgG antibody.

20. The monoclonal antibody of claim 19, wherein the full-length IgG antibody is a full-length IgG antibody of human IgG1, IgG2, IgG3, or IgG4 isotype.

21. The monoclonal antibody of claim 20, wherein the antibody comprises deletions of 446G and 447K in the CH3 region according to the EU numbering scheme for amino acids of antibodies.

22. The monoclonal antibody of claim 20, wherein the antibody comprises L234A and L235A mutations according to the EU numbering scheme for amino acids of antibodies.

23. The monoclonal antibody of claim 20, wherein the antibody comprises M252Y, S254T, T256E mutations according to the EU numbering scheme for amino acids of antibodies.

24. The monoclonal antibody of claim 1, comprising a light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, or SEQ ID NO:

60.

25. The monoclonal antibody of claim 1, comprising a heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, or SEQ ID NO:

74.

26. The monoclonal antibody of claim 1, comprising: (i)(a) a light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, or SEQ ID NO: 60, and (b) a heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, or SEQ ID NO:

74.

27. The monoclonal antibody according to claim 26, comprising: (i)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 61; or (ii) (a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 62; or (iii) (a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 63; or (iv) (a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 64; or (v)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 65; or (vi)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 66; or (vii)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 67; or (viii)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 68; or (ix)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 69; or (x)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 70; or (xi)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 57, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 71; or (xii)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 58, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 72; or (xiii)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 59, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 73; or (xiv)(a) a light chain comprising the amino acid sequence of SEQ ID NO: 60, and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO:

74.

28. A nucleic acid encoding the antibody or antigen-binding fragment thereof according to any one of claims 1 to 27.

29. The nucleic acid of claim 28, wherein the nucleic acid is DNA.

30. An expression vector comprising the nucleic acid according to any one of claims 28-29.

31. A method for producing a host cell for producing the antibody or antigen-binding fragment thereof according to any one of claims 1 to 27, comprising transforming the cell with the vector according to claim 31.

32. A host cell for producing the antibody or antigen-binding fragment thereof according to any one of claims 1-27, comprising a nucleic acid according to any one of claims 28-29.

33. A method for producing the antibody or antigen-binding fragment thereof according to any one of claims 1 to 27, comprising culturing the host cell according to claim 32 in a growth medium under conditions sufficient to produce the antibody, followed by isolating and purifying the resulting antibody.

34. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof according to any one of claims 1 to 27 in combination with one or more pharmaceutically acceptable excipients.

35. The pharmaceutical composition of claim 34, comprising the antibody or antigen-binding fragment thereof according to any one of claims 1 to 27 and at least one other therapeutically active compound.

36. The pharmaceutical composition of claim 35, wherein the other therapeutically active compound is an antibody, a small molecule, a hormonal therapeutic agent, or any combination thereof.

37. The pharmaceutical composition according to any one of claims 34 or 35, for use in treating an L1A-mediated disease or condition.

38. The pharmaceutical composition of claim 37, wherein the TL1A-mediated disease or condition is selected from the group consisting of: Crohn's disease, ulcerative colitis, bronchial asthma, rheumatoid arthritis, systemic lupus erythematosus, psoriasis, ankylosing spondylitis, systemic sclerosis-associated interstitial lung disease (SSc-ILD), intestinal fibrosis, liver fibrosis, or fibrotic lung disease.

39. A method for treating a TL1A-mediated disease or condition comprising administering to a subject in need of such treatment the antibody or antigen-binding fragment thereof according to any one of claims 1-27 or the pharmaceutical composition according to any one of claims 34-36.

40. The method of claim 39 comprising the antibody or antigen-binding fragment thereof according to any one of claims 1-27 or the pharmaceutical composition according to any one of claims 34-35 and at least one other therapeutically active compound.

41. The method for treating a disease or condition according to claim 40, wherein the other therapeutically active compound is an antibody, a small molecule, a hormonal therapeutic agent, or any combination thereof.

42. The method for treating a disease or condition according to any one of claims 39 to 41, wherein the TL1A-mediated disease or condition is selected from the group consisting of: Crohn's disease, ulcerative colitis, bronchial asthma, rheumatoid arthritis, systemic lupus erythematosus, psoriasis, ankylosing spondylitis, systemic sclerosis-associated interstitial lung disease (SSc-ILD), intestinal fibrosis, liver fibrosis, or fibrotic lung disease.

43. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1-27 or the pharmaceutical composition according to any one of claims 34-35 for treating a TL1A-mediated disease or disorder.

44. Use according to claim 43 of an antibody or antigen-binding fragment thereof according to any one of claims 1 to 27 or a pharmaceutical composition according to any one of claims 34 to 35 and at least one other therapeutically active compound for treating a TL1A-mediated disease or disorder.

45. The use according to claim 44, wherein the other therapeutically active compound is an antibody, a small molecule, a hormonal therapeutic agent or any combination thereof.

46. ​​The use according to any one of claims 43 to 45, wherein the TL1A-mediated disease or condition is selected from: Crohn's disease, ulcerative colitis, bronchial asthma, rheumatoid arthritis, systemic lupus erythematosus, psoriasis, ankylosing spondylitis, systemic sclerosis-associated interstitial lung disease (SSc-ILD), intestinal fibrosis, liver fibrosis or fibrotic lung disease.

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