Antibodies specifically binding to human DR3 and uses thereof

Abstract

The present invention relates to an antibody specifically binding to human DR3 capable of specifically binding to a specific epitope of DR3 and selectively acting on regulatory T cells, thereby minimizing the activity of CD4 + T cells and CD8 + T cells exhibiting effector activity while strongly activating only the regulatory T cells, and a use thereof. The antibody has an excellent effect of preventing or treating graft versus host disease, autoimmune disease, and type 2 diabetes mellitus by induction of such selective activity on regulatory T cells, and thus can be usefully used as a therapeutic agent for various diseases related thereto.

Description

[0001] This application claims priority to and the benefit of Korean Patent Application Nos. 10-2022-0074078 and 10-2023-0077023 filed in the Korean Intellectual Property Office on June 17, 2022 and June 15, 2023, respectively, and all contents disclosed in the specifications and drawings of these applications are incorporated into this application. Technical Field

[0002] The present invention relates to an antibody specifically binding to human DR3 and uses thereof. Background Art

[0003] Death receptor 3 (DR3, tumor necrosis factor receptor superfamily 25, TNFRSF25) is one of the TNFRSF members. DR3 consists of 417 amino acids, has a molecular weight of 47 kDa, and is composed of three homotrimers. Although DR3 is highly expressed in regulatory T cells (Treg) under normal conditions, the expression of DR3 also increases when conventional T cells are activated. In addition, DR3 is known to be expressed in immune cells (such as NK cells, ILCs, B cells, monocytes) and non-immune cells (such as osteocytes and endothelial colony-forming cells).

[0004] Tumor necrosis factor-like ligand 1A (TL1A) is a ligand for DR3, which is mainly expressed in activated T cells, dendritic cells, macrophages, monocytes and endothelial cells. TL1A activates NF-κB signaling by binding to DR3, thereby functioning as a co-stimulatory factor for T cells. According to previously reported papers, the use of DR3 agonists can induce selective activation of regulatory T cells. In this case, DR3 agonists are one of the methods for activating regulatory T cells, and only selectively activate regulatory T cells, which is different from IL-2, which activates effector T cells in addition to activating regulatory T cells. The activation of effector T cells may be a major problem that hinders treatment when treating various inflammatory diseases.

[0005] DR3 agonists can be TL1A, which is a ligand for DR3, or agonistic anti-DR3 antibodies. However, since TL1A has a decoy receptor (DcR3), which is a natural inhibitor and natural inhibitors of cytokines are usually regulated by negative feedback, the method of inducing DR3 activation using TL1A makes it difficult to effectively deliver drugs, and the characteristics of TL1A as a trimeric cytokine may increase the difficulty of the new drug development process. On the other hand, the use of agonistic anti-DR3 antibodies can avoid the negative feedback of DcR3 and induce effective DR3 activation.

[0006] At the same time, since the development of the first monoclonal antibody in 1975, research on the clinical applicability of antibody drugs has been ongoing, and research on the mechanism of action (i.e., the mechanism by which specific biological responses can be very powerfully and selectively controlled by utilizing the properties of antibodies with strong binding affinity and high binding specificity for antigens) is the fundamental core of developing the potential of antibody drugs.

[0007] In particular, antibody drugs show relatively fewer side effects and excellent therapeutic effects due to their high binding specificity and stability in the human body compared to existing therapeutic agents based on chemically synthesized substances. Therefore, the field of therapeutic agent development using antibodies has attracted attention as a core field for the development of next-generation new drugs.

[0008] However, antibody drugs that specifically bind to DR3 to activate regulatory T cells and have excellent therapeutic effects on graft-versus-host disease, autoimmune diseases, or type II diabetes have not yet been developed. Summary of the invention

[0009] [Technical issues]

[0010] The present invention aims to provide an anti-DR3 antibody or an antigen-binding fragment thereof, wherein the anti-DR3 antibody or the antigen-binding fragment thereof specifically binds to death receptor 3 (DR3).

[0011] The present invention also aims to provide a nucleic acid sequence encoding the antibody or antigen-binding fragment.

[0012] The present invention also aims to provide a vector comprising the nucleic acid sequence.

[0013] The present invention also aims to provide a transgenic organism transformed with the vector.

[0014] The present invention also aims to provide a pharmaceutical composition for preventing or treating graft-versus-host disease, autoimmune diseases or type II diabetes, wherein the pharmaceutical composition comprises an anti-DR3 antibody or an antigen-binding fragment thereof that specifically binds to death receptor 3 (DR3) as an active ingredient.

[0015] The present invention also aims to provide a kit for preventing or treating graft-versus-host disease, autoimmune diseases or type II diabetes, the kit comprising the carrier or the composition and instructions for use.

[0016] However, the technical problems to be solved by the present invention are not limited to the above-mentioned technical problems, and a person skilled in the art in the art to which the present invention belongs will clearly understand other unmentioned technical problems from the following description.

[0017] [Technical solution]

[0018] One aspect of the present invention provides an anti-DR3 antibody or an antigen-binding fragment thereof, which specifically binds to death receptor 3 (DR3).

[0019] Another aspect of the present invention provides a nucleic acid sequence encoding the antibody or antigen-binding fragment.

[0020] Another aspect of the present invention provides a vector comprising the nucleic acid sequence.

[0021] Another aspect of the present invention provides a transgenic organism transformed with the vector.

[0022] Another aspect of the present invention provides a pharmaceutical composition for preventing or treating graft-versus-host disease, autoimmune diseases or type II diabetes, the pharmaceutical composition comprising an anti-DR3 antibody or an antigen-binding fragment thereof that specifically binds to death receptor 3 (DR3) as an active ingredient.

[0023] Another aspect of the present invention provides a kit for preventing or treating graft-versus-host disease, autoimmune diseases or type II diabetes, the kit comprising the vector or the composition and instructions for use.

[0024] In an exemplary embodiment of the present invention, the antibody or antigen-binding fragment may be any one selected from the group consisting of, but not limited to:

[0025] i) an antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a heavy chain CDR 1 as shown in the amino acid sequence of SEQ ID NO: 1, a heavy chain CDR 2 as shown in the amino acid sequence of SEQ ID NO: 2, and a heavy chain CDR 3 as shown in the amino acid sequence of SEQ ID NO: 3; and the light chain variable region comprises a light chain CDR 1 as shown in the amino acid sequence of SEQ ID NO: 4, a light chain CDR 2 as shown in the amino acid sequence of SEQ ID NO: 5, and a light chain CDR 3 as shown in the amino acid sequence of SEQ ID NO: 6;

[0026] ii) an antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the heavy chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 7, the heavy chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 8, and the heavy chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 9; and the light chain variable region comprises the light chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 10, the light chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 11, and the light chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 12;

[0027] iii) an antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a heavy chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 13, a heavy chain CDR2 set forth in the amino acid sequence of SEQ ID NO: 14, and a heavy chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 15; and the light chain variable region comprises a light chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 16, a light chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 17, and a light chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 18;

[0028] iv) an antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a heavy chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 19, a heavy chain CDR2 set forth in the amino acid sequence of SEQ ID NO: 20, and a heavy chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 21; and the light chain variable region comprises a light chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 22, a light chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 23, and a light chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 24; and

[0029] v) an antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising a heavy chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 25, a heavy chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 26, and a heavy chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 27; the light chain variable region comprising a light chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 28, a light chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 29, and a light chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 30.

[0030] In an exemplary embodiment of the present invention, the antibody or antigen-binding fragment thereof may be any one selected from the group consisting of, but not limited thereto:

[0031] i) an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 63 and a light chain variable region represented by the amino acid sequence of SEQ ID NO: 64;

[0032] ii) an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 67 and a light chain variable region represented by the amino acid sequence of SEQ ID NO: 68;

[0033] iii) an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 71 and a light chain variable region represented by the amino acid sequence of SEQ ID NO: 72;

[0034] iv) an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 75 and a light chain variable region represented by the amino acid sequence of SEQ ID NO: 76; and

[0035] v) an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 79 and a light chain variable region represented by the amino acid sequence of SEQ ID NO: 80.

[0036] In an exemplary embodiment of the present invention, the antibody or antigen-binding fragment thereof may include a heavy chain represented by the amino acid sequence of SEQ ID NO: 83 and a light chain represented by the amino acid sequence of SEQ ID NO: 84, but is not limited thereto.

[0037] In an exemplary embodiment of the present invention, the antibody or antigen-binding fragment may include Fab, but is not limited thereto.

[0038] In an exemplary embodiment of the present invention, the antibody or antigen-binding fragment may be of IgG1 subclass or IgG4 subclass, but is not limited thereto.

[0039] In an exemplary embodiment of the present invention, the antigen-binding fragment may be any one selected from the group consisting of Fv, Fab, F(ab')2, Fab', dsFv, scFv, sc(Fv)2 and diabody, but is not limited thereto.

[0040] In an exemplary embodiment of the present invention, the antibody or antigen-binding fragment of iii), iv) and v) can specifically bind to DR3 competitively with TL1A, but is not limited thereto.

[0041] In an exemplary embodiment of the present invention, the characteristics of the antibody or antigen-binding fragment may be any one or more of the following, but are not limited to:

[0042] Selectively activate regulatory T cells or enhance their survival; and

[0043] Activation of NF-κB.

[0044] In an exemplary embodiment of the present invention, the autoimmune disease may be any one or more selected from the group consisting of: an autoimmune disease caused by graft-versus-host disease, hemophagocytic lymphohistiocytosis, systemic lupus erythematosus, Kikuchi's disease, adult onset Still's disease, Behcet's disease, IgG4-related disease, type I diabetes, systemic sclerosis, psoriasis, multiple sclerosis and Graves hyperthyroidism, but is not limited thereto.

[0045] Another aspect of the present invention provides a method for preventing or treating graft-versus-host disease, autoimmune diseases or type II diabetes, the method comprising administering to a subject in need thereof an anti-DR3 antibody or an antigen-binding fragment thereof, or a composition comprising the same as an active ingredient, wherein the anti-DR3 antibody or the antigen-binding fragment thereof binds to death receptor 3 (DR3).

[0046] Another aspect of the present invention provides use of an anti-DR3 antibody or an antigen-binding fragment thereof, or a composition comprising the same as an active ingredient, for preventing or treating graft-versus-host disease, autoimmune diseases or type II diabetes, wherein the anti-DR3 antibody or the antigen-binding fragment thereof binds to death receptor 3 (DR3).

[0047] Another aspect of the present invention provides use of an anti-DR3 antibody or an antigen-binding fragment thereof, or a composition comprising the same as an active ingredient, in the preparation of a preparation for preventing or treating graft-versus-host disease, an autoimmune disease or type II diabetes, wherein the anti-DR3 antibody or the antigen-binding fragment thereof binds to death receptor 3 (DR3).

[0048] [Beneficial Effects]

[0049] An antibody that specifically binds to human DR3 and its use, which can minimize the activity of effector CD4+T cells and CD8+T cells while increasing the division or survival rate of regulatory T cells by specifically binding to a specific epitope of human DR3 and selectively acting on regulatory T cells. Due to this activity, the antibody has a very excellent preventive or therapeutic effect on related diseases, so the antibody can be effectively used as a therapeutic agent for graft-versus-host disease, autoimmune diseases or type II diabetes. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] Figure 1a and 1b Shown are experimental results of a method for discovering anti-DR3 scFv clones that specifically bind to DR3 by phage display.

[0051] Figures 2a to 2c The binding specificity and affinity of anti-DR3 monoclonal antibodies to DR3 are shown.

[0052] Figure 3a and 3b The results of confirming that the binding site of the anti-DR3 4-4 antibody to DR3 overlaps with the binding site of TL1A are shown.

[0053] Figure 4 The NF-κB activation levels of the anti-DR3 antibodies of the present invention are shown.

[0054] Figure 5 The results of comparison of the affinity of the anti-DR3 4-4 antibody to DR3 according to IgG subclass are shown.

[0055] Figure 6 Shown are the results of comparative analysis of NF-κB activation levels of anti-DR3 4-4 antibodies according to IgG subclass.

[0056] Figure 7 The results show that the anti-DR3 4-4 IgG4 antibody improves the survival rate of regulatory T cells in the absence of TCR signal stimulation.

[0057] Figure 8 The modulatory effect of anti-DR3 4-4 IgG4 antibody on the division and survival of regulatory T cells according to the TCR signal strength is shown. DETAILED DESCRIPTION

[0058] In the present invention, anti-DR3 antibodies that bind to the natural form of DR3 were discovered through full human phage display and stable cell lines expressing DR3. The anti-DR3 antibodies of the present invention have been confirmed to act as agonists, activating T cells by specifically binding to DR3 expressed by human T cells by competing with TL1A. Further, by preparing antibodies against clone 4-4, which most strongly activates T cells among them, to obtain different isotypes (e.g., IgG1, IgG2, IgG4, etc.), the degree of activation of T cells by each isotype antibody was compared, and the anti-DR3 IgG4 antibody was confirmed to specifically bind to the natural form of DR3 and has the strongest activation effect on T cells. In addition, through these, it was confirmed that the anti-DR34-4IgG4 antibody, which was identified as the most excellent antibody, increased the survival rate of regulatory T cells in the absence of TCR signal stimulation, and effectively increased the division or survival rate of regulatory T cells in the presence of TCR signal stimulation of a certain intensity or higher, thereby completing the present invention.

[0059] The present invention provides an anti-DR3 antibody or an antigen-binding fragment thereof, wherein the anti-DR3 antibody or the antigen-binding fragment thereof specifically binds to death receptor 3 (DR3).

[0060] In the present invention, DR3 (TNFRSF25, TNF receptor superfamily member 25, death receptor 3) is a member of the tumor necrosis factor receptor superfamily, which is expressed at high levels in regulatory T cells, but can also be induced to express in existing T cells through TCR stimulation. In the present invention, DR3 may include DR3, hDR3, etc., but is not limited thereto. As confirmed in mice using anti-DR3 antibody agonists, DR3 signaling can promote the activation and proliferation of regulatory T cells under normal conditions, and only a small amount of DR3 can induce the proliferation of regulatory T cells rather than the proliferation of non-target immune cell populations, which is a key difference from the IL-2 complex (IL-2 also promotes the division of CD4 T cells and CD8 T cells). Therefore, since the division of conventional T cells (Tconv) can be a major obstacle to the treatment of various inflammatory diseases, anti-DR3 antibodies can be used as a new method for selectively activating regulatory T cells.

[0061] In an exemplary embodiment of the present invention, the antibody or antigen-binding fragment may be any one selected from the group consisting of, but not limited to:

[0062] i) an antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a heavy chain CDR 1 as shown in the amino acid sequence of SEQ ID NO: 1, a heavy chain CDR 2 as shown in the amino acid sequence of SEQ ID NO: 2, and a heavy chain CDR 3 as shown in the amino acid sequence of SEQ ID NO: 3; and the light chain variable region comprises a light chain CDR 1 as shown in the amino acid sequence of SEQ ID NO: 4, a light chain CDR 2 as shown in the amino acid sequence of SEQ ID NO: 5, and a light chain CDR 3 as shown in the amino acid sequence of SEQ ID NO: 6;

[0063] ii) an antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the heavy chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 7, the heavy chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 8, and the heavy chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 9; and the light chain variable region comprises the light chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 10, the light chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 11, and the light chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 12;

[0064] iii) an antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a heavy chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 13, a heavy chain CDR2 set forth in the amino acid sequence of SEQ ID NO: 14, and a heavy chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 15; and the light chain variable region comprises a light chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 16, a light chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 17, and a light chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 18;

[0065] iv) an antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a heavy chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 19, a heavy chain CDR2 set forth in the amino acid sequence of SEQ ID NO: 20, and a heavy chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 21; and the light chain variable region comprises a light chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 22, a light chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 23, and a light chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 24; and

[0066] v) an antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising a heavy chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 25, a heavy chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 26, and a heavy chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 27; the light chain variable region comprising a light chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 28, a light chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 29, and a light chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 30.

[0067] In an exemplary embodiment of the present invention, the antibody or antigen-binding fragment thereof may be any one selected from the group consisting of, but not limited to:

[0068] i) an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 63 and a light chain variable region represented by the amino acid sequence of SEQ ID NO: 64;

[0069] ii) an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 67 and a light chain variable region represented by the amino acid sequence of SEQ ID NO: 68;

[0070] iii) an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 71 and a light chain variable region represented by the amino acid sequence of SEQ ID NO: 72;

[0071] iv) an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 75 and a light chain variable region represented by the amino acid sequence of SEQ ID NO: 76; and

[0072] v) an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 79 and a light chain variable region represented by the amino acid sequence of SEQ ID NO: 80.

[0073] In the present invention, the terms "percent identity", "sequence identity", "percent similarity", "sequence similarity", "percent sequence identity" and the like used herein in connection with amino acid sequences and / or nucleic acid sequences may refer to a measure determined by comparing the degree of similarity of two sequences based on sequence alignment, which maximizes the similarity between the aligned amino acid residues or nucleotides, and is a function of the number of identical or similar residues or nucleotides in the sequence alignment, the total number of residues or nucleotides, and the presence and length of gaps, but is not limited thereto.

[0074] A portion of a polynucleotide or polypeptide sequence may comprise additions or deletions (i.e., gaps) compared to a reference sequence (which does not comprise additions or deletions) to achieve optimal alignment of the two sequences. The percentage is calculated by determining the number of positions indicated by the same nucleobase or amino acid residue in the two sequences to calculate the number of matching positions, dividing the number of matching positions in the comparison window by the total number of positions, and multiplying the result by 100 to calculate the sequence identity percentage.

[0075] The term "identical" or "percentage of identity" in relation to two or more nucleic acid or polypeptide sequences refers to two or more sequences or subsequences that are identical or have a specified percentage of identical amino acid residues or nucleotides (i.e., about 60% identity, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or even higher identity to a specific region when compared and aligned over a comparison window or indicated region for maximum correspondence), as determined using BLAST or BLAST 2.0 sequence comparison algorithms and using the default parameters described below, or as determined by manual alignment and visual inspection (see, e.g., NCBI website http: / / www.ncbi.nlm.nih.gov / BLAST / , etc.). Such sequences are hereinafter referred to as "substantially identical."

[0076] In the present invention, the amino acid sequence or base sequence may include a base sequence having at least 70% sequence identity, more preferably at least 80%, even more preferably at least 90%, and most preferably at least 95% sequence identity with the amino acid sequence or base sequence represented by each sequence number. For example, the amino acid sequence or base sequence may include a sequence having 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence homology, but the percentage is not limited thereto.

[0077] In the present invention, naturally occurring antibodies are glycoproteins comprising at least two heavy (H) chains and two light (L) chains connected by interchain disulfide bonds. Each heavy chain consists of a heavy chain variable region (VH) and a heavy chain constant region. The heavy chain constant region consists of three domains: CH1, CH2, and CH3. Each light chain consists of a light chain variable region (abbreviated as VL) and a light chain constant region. The light chain constant region consists of the domain CL.

[0078] The VH and VL regions can be further subdivided into regions of high variability called complementarity determining regions (CDRs), which are separated by more conserved regions called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs, which are arranged in the following order from amino terminus to carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens.

[0079] In the present invention, antibodies can be derived from a variety of species, such as mice, rats, rabbits and guinea pigs, but species are not limited thereto. In addition, antibodies include chimeric antibodies having an antibody constant region (preferred) from one species and an antigen binding site from another species. In addition, antibodies include antibodies having an antigen binding site of an antibody from a non-human species and antibodies having an original constant region and framework region. The antibodies described herein can be monoclonal antibodies, i.e. antibody molecules having a single molecular component. Systems and immunological procedures and techniques for preparing and isolating hybridomas secreting monoclonal antibodies are known in the art.

[0080] As used herein, "antigen" refers to a substance that contains a binding site (epitope) and induces and / or produces an immune response to the epitope. Antigens may include peptides, proteins, glycoproteins, polysaccharides and lipids, parts thereof and combinations thereof, but are not limited thereto. Non-limiting exemplary antigens include tumor antigens or pathogenic antigens. In addition, the term "antigen" may refer to a molecule that triggers an immune response. This immune response may involve the production of antibodies or the activation of specific immunologically active cells or both. It will be appreciated by those skilled in the art that any macromolecule (including virtually all proteins or peptides) may be used as an antigen.

[0081] As used herein, "binding site" or "epitope" may refer to an antigenic determinant in a molecule, i.e., a portion of a molecule that is recognized by the immune system (e.g., by an antibody). In the present invention, the epitope may include a portion of DR3 that binds to TL1A or a portion of DR3 that binds to an anti-DR3 antibody, but is not limited thereto.

[0082] In an exemplary embodiment of the present invention, the antibody or antigen-binding fragment thereof may include a heavy chain shown in the amino acid sequence of SEQ ID NO: 83 and a light chain shown in the amino acid sequence of SEQ ID NO: 84, but is not limited thereto.

[0083] As a result of preparing the 4-4IgG4 antibody confirmed to be most effective in the application to graft-versus-host disease, autoimmune diseases or type II diabetes in the present invention, in an exemplary embodiment of the present invention, the antibody or antigen-binding fragment may include Fab, but is not limited thereto.

[0084] Therapeutic antibodies are generally immunoglobulin G (IgG), and IgG is divided into four subclasses (IgG1, IgG2, IgG3, and IgG4). Since the IgG subclasses have different effector functions, it is very important to select the appropriate IgG subclass according to the indications of the therapeutic antibody. In this case, it was confirmed that not only the effector function of the antibody is different, but also the binding affinity and activation function of the antibody to the target antigen are different according to the IgG class. In addition, the strength of the binding affinity of the IgG subclass will also vary depending on the type of antigen antibody.

[0085] As a result of confirming that the 4-4 antibody, which was confirmed to be the most effective in the present invention, was confirmed to be the most functionally optimal isoform, in an exemplary embodiment of the present invention, the antibody or antigen-binding fragment may be of the IgG4 subclass, but is not limited thereto.

[0086] In an exemplary embodiment of the present invention, the antibody or antigen-binding fragment may be of IgG1 or IgG4 subclass, but is not limited thereto.

[0087] As used herein, "antigen-binding fragment" refers to an antibody fragment that retains specific antigen binding ability, and refers to one or more fragments of an antibody, including, for example: (i) Fab fragment, a monovalent fragment consisting of VL, VH, CL and CH domains; (ii) F(ab')2 fragment, a bivalent fragment comprising two Fab fragments connected by a disulfide bond at the hinge region; (iii) Fd fragment, consisting of VH and CH domains; (iv) Fv fragment, consisting of VL and VH domains of a single arm of an antibody; (v) a single domain antibody fragment consisting of a VH domain; (vi) an isolated complementary determining region (CDR); and (vii) a binding fragment comprising a combination of two or more isolated CDRs, the two or more isolated CDRs being optionally connected by a synthetic linker. There are also single-chain antibodies (or single-chain Fv (scFv)) formed using recombinant methods to connect the two domains (VL and VH) of the FV segment by a synthetic linker.

[0088] In an exemplary embodiment of the present invention, the antigen-binding fragment may be any one of the group consisting of Fv, Fab, F(ab')2, Fab', dsFv, scFv, sc(Fv)2 and diabody, but is not limited thereto.

[0089] In the present invention, since an anti-DR3 antibody having a binding site overlapping with TL1A, a ligand that activates human DR3, was discovered through epitope mapping, in order to develop an agonistic anti-DR3 antibody for activating the target antigen human DR3, the antibody or antigen-binding fragment of the present invention may have a DR3 binding epitope that is identical or overlapping with TL1A. In this case, the antibody or antigen-binding fragment of the present invention may competitively and specifically bind to DR3 to induce activation of regulatory T cells, etc., but is not limited thereto. In addition, when TL1A and the epitope that binds to DR3 do not overlap, the epitope may bind to DR3 together with TL1A to activate DR3 excellently and non-competitively. Therefore, the anti-DR3 antibody of the present invention may activate the function of DR3 by competitively and / or non-competitively binding to DR3 with TL1A, thereby exerting the effect of preventing or treating graft-versus-host disease, autoimmune diseases, or type II diabetes.

[0090] In an exemplary embodiment of the present invention, the antibodies or antigen-binding fragments of iii), iv) and v) may specifically bind to DR3 competitively with TL1A, but are not limited thereto. That is, the binding site (epitope) of the antibodies or antigen-binding fragments of iii), iv) and v) for DR3 may overlap with the binding site of TL1A for DR3, but are not limited thereto.

[0091] In an exemplary embodiment of the present invention, the characteristics of the antibody or antigen-binding fragment may be any one or more of the following, but are not limited to:

[0092] Selectively activate regulatory T cells or enhance regulatory T cell survival; and

[0093] Activates NF-κB.

[0094] In the present invention, the anti-DR3 antibody may exhibit an activation efficiency of about 10% to 80% for CD3+CD4+NF-κB, and may exhibit an activation efficiency of about 1% to 60% for CD3+CD8+NF-κB, depending on the concentration, but is not limited thereto. In particular, the anti-DR3 4-4IgG1 antibody of the present invention may increase the activation efficiency of CD3+CD4+NFκB and CD3+CD8+NF-κB to about 80% and 60%, respectively, by inducing a phosphorylation process, depending on the antibody treatment concentration, to improve the activation efficiency of NF-κB cells, but is not limited thereto. In the present invention, the anti-DR3 antibody may exhibit an activation efficiency of greater than 10 to 100%, greater than 10 to 99%, greater than 10 to 98%, greater than 10 to 97%, greater than 10 to 96%, greater than 10 to 95%, greater than 10 to 94%, greater than 10 to 93%, greater than 10 to 92%, greater than 10 to 91%, greater than 10 to 90%, greater than 10 to 89%, greater than 10 to 88%, greater than 10 to 87%, greater than 10 to 86%, greater than 10 to 85%, greater than 10 to 84%, greater than 10 to 83%, greater than 10 to 82% or greater than 10 to 81% for CD3+CD4+NF-κB, and the activation efficiency may preferably be greater than 10 to 80%, but is not limited thereto. In addition, the anti-DR3 antibody may exhibit an activation efficiency of greater than 0 to 100%, greater than 0 to 95%, greater than 0 to 90%, greater than 0 to 85%, greater than 0 to 80%, greater than 0 to 75%, greater than 0 to 70%, greater than 0 to 69%, greater than 0 to 68%, greater than 0 to 67%, greater than 0 to 66%, greater than 0 to 65%, greater than 0 to 64%, greater than 0 to 63%, greater than 0 to 62%, or greater than 0 to 61%, and the activation efficiency may preferably be greater than 0 to 60%, but is not limited thereto.

[0095] In the present invention, anti-DR3 antibodies can induce strong expansion of regulatory T cells in vivo. For example, the number and percentage of regulatory T cells can be increased in lymph nodes and spleen, and can selectively express regulatory T cells, but is not limited thereto.

[0096] In the present invention, the anti-DR3 antibody exhibits a significantly lower binding dissociation constant for DR3 than TL1A, which is a ligand of DR3, and thus can exhibit excellent binding strength. Compared with TL1A, the anti-DR3 antibody of the present invention can exhibit a binding dissociation constant of greater than about 2 times to about 10,000 times, greater than about 2 times to about 9,800 times, greater than about 2 times to about 9,600 times, greater than about 2 times to about 9,400 times, greater than about 2 times to about 9,200 times, greater than about 4 times to about 10,000 times, greater than about 4 times to about 9,800 times, greater than about 4 times to about 9,600 times, greater than about 4 times to about 9,400 times, greater than about 4 times to about 9,200 times, greater than about 6 times to about 10,000 times, greater than about 6 times to about 9,800 times, greater than about 6 times to about 9,600 times, greater than about 6 times to about 9,400 times, greater than about 6 times to about 9,200 times, greater than about 8 times to about 1 The bonding strength may be greater than about 12 times to about 10,000 times, greater than about 8 times to about 9,800 times, greater than about 8 times to about 9,600 times, greater than about 8 times to about 9,400 times, greater than about 8 times to about 9,200 times, greater than about 10 times to about 10,000 times, greater than about 10 times to about 9,800 times, greater than about 10 times to about 9,600 times, greater than about 10 times to about 9,400 times, greater than about 10 times to about 9,200 times, greater than about 12 times to about 10,000 times, greater than about 12 times to about 9,800 times, greater than about 12 times to about 9,600 times, greater than about 12 times to about 9,400 times, and may exhibit a bonding strength preferably greater than about 12 times to about 9,200 times, but the bonding strength is not limited thereto.

[0097] The present invention provides a nucleic acid sequence encoding the antibody or antigen-binding fragment.

[0098] As used herein, "genome" may refer to the genetic material (e.g., chromosomes) of an organism, and "nucleic acid sequence" or "gene" may refer to a nucleic acid (e.g., DNA or RNA) sequence that contains the coding sequence required for producing a polypeptide or precursor (e.g., proinsulin), but is not limited thereto.

[0099] The "nucleotide sequence" or "base sequence" in the present invention refers to any nucleotide sequence (e.g., RNA or DNA), and the manipulation thereof by a person of ordinary skill in the art may also be considered preferred for any reason (e.g., to treat a disease or improve a property, to express a protein of the present invention in a host cell, or to convey the expression of a ribozyme). Such nucleotide sequences may include, but are not limited to, coding sequences of structural genes (e.g., reporter genes, selectable marker genes, oncogenes, drug resistance genes, growth factors, etc.) and non-coding regulatory sequences that do not encode mRNA or protein products (e.g., promoter sequences, polyadenylation sequences, termination sequences, enhancer sequences, etc.).

[0100] The present invention provides a vector comprising the nucleic acid sequence.

[0101] The term "vector" as used herein refers to any genetic element capable of replicating and transferring genetic sequences between cells when linked to appropriate regulatory elements, such as plasmids, phages, transposons, cosmids, chromosomes, viruses and virions.

[0102] As used herein, "recombinant expression vector" refers to bacterial plasmids, bacteriophages, yeast plasmids, plant cell viruses, mammalian cell viruses or other vectors. Generally speaking, any plasmid or vector can be used as long as it is replicable and stable in the host.

[0103] Recombinant expression vectors and expression vectors containing appropriate transcription / translation control signals can be constructed by methods well known to those skilled in the art, including in vitro recombinant DNA technology, DNA synthesis technology, and in vivo recombination technology.

[0104] In addition, as a vector for recombination, both viral vectors and non-viral vectors can be used. For example, as a viral vector, a lentivirus, a retrovirus, an adenovirus, a herpes virus, or an avipox virus vector can be used, but the viral vector is not limited thereto.

[0105] In addition, the vector may also contain a tag gene for increasing the yield of the recombinant protein, a tag gene for maintaining the structural stability of the recombinant protein, a tag gene for easily isolating the recombinant protein, and a selective marker gene (e.g., an antibiotic resistance gene for selecting transgenic organisms), etc., but is not limited thereto.

[0106] The present invention provides a transgenic organism transformed with the vector.

[0107] As used herein, "transformation" refers to the process of changing the genetic characteristics of a living organism by DNA injection, and "transgenic organism" refers to an organism prepared by injecting external genes using molecular genetic methods, preferably a cell or organism transformed by the recombinant expression vector of the present invention, but is not limited thereto.

[0108] In some cases, the organism may include, but is not limited to, any living organism, such as a microorganism, a eukaryotic cell, an insect, an animal or a plant, and preferably Escherichia coli, Salmonella, Bacillus, yeast, animal cells, mice, rats, dogs, monkeys, pigs, horses, cattle, Agrobacterium tumefaciens, plants, etc., but are not limited thereto.

[0109] Transgenic organisms can be prepared by methods such as transformation, transfection, Agrobacterium-mediated transformation, particle gun bombardment, ultrasound, electroporation, and polyethylene glycol (PEG)-mediated transformation, but the method is not limited thereto. In addition, by the method of introducing transgenic organisms into cells, the transgenic organisms can be introduced into cells with a delivery agent (including G-fectin, Mirus TransIT-TKO lipophilic agent, lipofectin, lipofectamine, cellfectin, cationic phospholipid nanoparticles, cationic polymers, cationic micelles, cationic emulsions or liposomes) or the transgenic organisms can be conjugated with a biocompatible polymer (e.g., polyethylene glycol) to increase intracellular uptake, but the method is not limited thereto.

[0110] When the vector of the present invention is transformed into eukaryotic cells, yeast (Saccharomyces cerevisiae), insect cells, human cells (e.g., CHO cell lines (Chinese hamster ovary), W138, BHK, COS-7, 293, HepG2, 3T3, RIN and MDCK cell lines), plant cells, etc. can be used as host cells, and preferably Agrobacterium can be used.

[0111] When the host cell is a prokaryotic cell, the method for introducing the vector of the present invention into the host cell can be carried out by the CaCl2 method, the Hanahan method (Hanahan, D., J. Mol. Biol., 166: 557-580 (1983)), the electroporation method, etc. In addition, when the host cell is a eukaryotic cell, the vector can be injected into the host cell by microinjection, calcium phosphate precipitation, electroporation, liposome-mediated transfection, DEAE-dextran treatment, gene bombardment, etc.

[0112] The present invention provides a pharmaceutical composition for preventing or treating graft-versus-host disease, autoimmune diseases or type II diabetes, wherein the pharmaceutical composition comprises an anti-DR3 antibody or an antigen-binding fragment thereof that specifically binds to death receptor 3 (DR3) as an active ingredient.

[0113] In an exemplary embodiment of the present invention, the antibody or antigen-binding fragment thereof may include a heavy chain represented by the amino acid sequence of SEQ ID NO: 83 and a light chain represented by the amino acid sequence of SEQ ID NO: 84, but is not limited thereto.

[0114] In an exemplary embodiment of the present invention, the autoimmune disease may be any one or more selected from the group consisting of: an autoimmune disease caused by graft-versus-host disease, hemophagocytic lymphohistiocytosis, systemic lupus erythematosus, Kikuchi's disease, adult-onset Still's disease, Behcet's disease, IgG4-related disease, type I diabetes, systemic sclerosis, psoriasis, multiple sclerosis and Graves' hyperthyroidism, but is not limited thereto.

[0115] The pharmaceutical composition of the present invention may also include suitable carriers, adjuvants and diluents commonly used in the preparation of pharmaceutical compositions. The adjuvant may be, for example, one or more selected from the group consisting of diluents, binders, disintegrants, lubricants, adsorbents, wetting agents, film coating materials and controlled release additives.

[0116] The pharmaceutical composition of the present invention can be formulated into an external preparation, such as a powder, granule, sustained-release granule, enteric-coated granule, liquid, eyewash, elixir, emulsion, suspension, spirit, lozenge, aromatic water, lemonade, tablet, sustained-release tablet, enteric-coated tablet, sublingual tablet, hard capsule, soft capsule, sustained-release capsule, enteric-coated capsule, pill, tincture, soft extract, dry extract, fluid extract, injection, capsule, perfusion, plaster, lotion, paste, spray, inhalant, patch, sterile injection, aerosol. The external preparation can have dosage forms such as cream, gel, patch, spray, ointment, plaster, lotion, liniment, paste or cataplasm.

[0117] Examples of carriers, excipients or diluents that may be included in the composition according to the present invention include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylparaben, propylparaben, talc, magnesium stearate and mineral oil.

[0118] When preparing a pharmaceutical composition, a commonly used diluent or excipient (such as a filler, an extender, a binder, a wetting agent, a disintegrant, and a surfactant) is used to prepare the pharmaceutical composition.

[0119] As additives for tablets, powders, granules, capsules, pills and lozenges according to the present invention, auxiliary materials such as corn starch, potato starch, wheat starch, lactose, sucrose, glucose, fructose, D-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, calcium hydrogen phosphate, calcium sulfate, sodium chloride, sodium bicarbonate, purified lanolin, microcrystalline cellulose, dextrin, sodium alginate, methylcellulose, sodium carboxymethylcellulose, kaolin, urea, colloidal silica, hydroxypropyl starch, hydroxypropyl methylcellulose (HPMC) 1928, HPMC 2208, HPMC 2906, HPMC 2910, propylene glycol, casein, calcium lactate or Primojel; binders such as gelatin, gum arabic, ethanol, agar powder, cellulose acetate phthalate, carboxymethyl cellulose, carboxymethyl cellulose calcium, glucose, purified water, sodium caseinate, glycerin, stearic acid, sodium carboxymethyl cellulose, sodium methyl cellulose, methyl cellulose, microcrystalline cellulose, dextrin, hydroxycellulose, hydroxypropyl starch, hydroxymethyl cellulose, purified shellac, starch, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyvinyl alcohol or polyvinyl pyrrolidone; disintegrants such as hydroxypropyl methyl cellulose, corn starch, agar powder, methyl cellulose, bentonite, hydroxypropyl starch, carboxymethyl cellulose sodium sorbitol, sodium alginate, calcium carboxymethylcellulose, calcium citrate, sodium lauryl sulfate, silicic anhydride, 1-hydroxypropyl cellulose, dextran, ion exchange resin, polyvinyl acetate, casein and gelatin treated with formaldehyde, alginic acid, amylose, guar gum, sodium bicarbonate, polyvinyl pyrrolidone, calcium phosphate, gelling starch, gum arabic, pullulan, pectin, sodium polyphosphate, ethyl cellulose, sucrose, magnesium aluminum silicate, D-sorbitol solution or light anhydrous silicic acid; and lubricants such as calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopodium powder, kaolin, vaseline, sodium stearate, cocoa butter, sodium salicylate, magnesium salicylate, polyethylene glycol 4000, PEG 6000, liquid paraffin, hydrogenated soybean oil (Lubri wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, polyethylene glycol (Macrogol), synthetic aluminum silicate, silicic anhydride, higher fatty acids, higher alcohols, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, starch, sodium chloride, sodium acetate, sodium oleate, dl-leucine or light anhydrous silicic acid.

[0120] As additives for the liquid preparation according to the present invention, water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, sucrose monostearate, polyoxyethylene sorbitan fatty acid esters (Tween esters), polyoxyethylene monoalkyl ethers, lanolin ethers, lanolin esters, acetic acid, hydrochloric acid, ammonia water, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamin, polyvinyl pyrrolidone, ethyl cellulose, sodium carboxymethyl cellulose, etc. can be used.

[0121] In the syrup according to the present invention, solutions of sucrose, other sugars or sweeteners etc. may be used, and flavoring agents, colorants, preservatives, stabilizers, suspending agents, emulsifiers, thickeners etc. may be used if necessary.

[0122] In the emulsion according to the present invention, purified water may be used, and an emulsifier, a preservative, a stabilizer, a fragrance, etc. may be used if necessary.

[0123] In the suspension according to the present invention, suspending agents such as gum arabic, gum tragacanth, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropyl methylcellulose, HPMC 1828, HPMC 2906 and HPMC 2910 may be used, and surfactants, preservatives, colorants and aromatics may be used if necessary.

[0124] The injection according to the present invention may include: distilled water for injection, 0.9% sodium chloride injection, Ringer's injection, glucose injection, glucose + sodium chloride injection, PEG, lactated Ringer's injection, ethanol, propylene glycol, non-volatile oils such as sesame oil, cottonseed oil, peanut oil, corn oil, ethyl oleate, isopropyl myristate or benzene benzoate; cosolvents such as sodium benzoate, sodium salicylate, sodium acetate, urea, ethyl carbamate, monoethylacetamide, phenylbutazone, propylene glycol, Tween, nijungtinateamide, methenamine or dimethylacetamide; buffers such as weak acids or their salts (acetic acid or sodium acetate). ), weak base or its salt (ammonia or ammonium acetate), organic compound, protein, albumin, peptone or gum; isotonic agent, such as sodium chloride; stabilizer, such as sodium bisulfite (NaHSO3), carbon dioxide gas, sodium pyrosulfite (Na2S2O5), sodium sulfite (Na2SO3), nitrogen (N2) or ethylenediaminetetraacetic acid; sulfating agent, such as 0.1% sodium hydrosulfide, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetic acid or sodium acetone bisulfite; analgesic, such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose or calcium gluconate, etc.; suspending agent, such as sodium carboxymethylcellulose, sodium alginate, Tween 80 or aluminum monostearate.

[0125] In the suppositories according to the invention, bases such as cocoa butter, lanolin, Witepsol, polyethylene glycol, glycerinated gelatin, methylcellulose, carboxymethylcellulose, a mixture of stearic acid and oleic acid, Subanal, cottonseed oil, peanut oil, palm oil, cocoa butter + cholesterol, lecithin, ranetwax, glyceryl monostearate, Tween or Span, Imhausen, monolen (propylene glycol monostearate, glycerol, mixed fatty acid glycerides (Adeps Solidus), Buytyrum Tego-G, Cebes Pharma 16, hexalide base 95, Cotomar, Hydrokote SP, S-70-XXA, S-70-XX75 (S-70-XX95), Hydrokote 25, Hydrokote 711, idropostal, Massa estrarium (A, AS, B, C, D, E, I, T), Massa-MF, Masupol, Masupol-15, Neosupostal-ene, Paramound-B, Suposhiro (OSI, OSIX, A, B, C, D, H, L), suppository base type IV (AB, B, A, BC, BBG, E, BGF, C, D, 299), Supostal (N, Es), Wecobee (W, R, S, M, Fs) and Tegester triglyceride base (TG-95, MA, 57).

[0126] Solid preparations for oral administration include tablets, pills, powders, granules or capsules, etc., which are prepared by mixing at least one auxiliary material (such as starch, calcium carbonate, sucrose or lactose, gelatin, etc.) with the extract. In addition, in addition to simple auxiliary materials, lubricants such as magnesium stearate and talc are also used.

[0127] Liquid preparations for oral administration correspond to suspensions, internal liquids, emulsions or syrups, etc., and in addition to commonly used simple diluents (such as water and liquid paraffin), liquid preparations can also contain a variety of adjuvants, such as wetting agents, sweeteners, aromatics and preservatives, etc. Examples of preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspending agents, emulsions, freeze-dried preparations and suppositories. As non-aqueous solvents and suspending agents, propylene glycol, polyethylene glycol, vegetable oils (such as olive oil), injectable esters (such as ethyl oleate), etc. can be used.

[0128] The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" refers to an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the level of the effective dose can be determined based on factors including the patient's disease type, the severity of the disease, the activity of the drug, sensitivity to the drug, administration time, administration route, excretion rate, treatment cycle, concurrently used drugs, and other factors well known in the medical field.

[0129] The pharmaceutical composition according to the present invention can be used as a single therapeutic agent or in combination with other therapeutic agents, can be used sequentially or simultaneously with the therapeutic agent of the related field, and can be used in a single dose or multiple doses. Taking all the aforementioned factors into consideration, it is important to use the composition in the minimum amount that can achieve the maximum effect without any side effects, and this amount can be easily determined by a person of ordinary skill in the art to which the present invention belongs.

[0130] The pharmaceutical composition of the present invention can be administered to a subject in need by various routes. All administration methods are contemplated, but the pharmaceutical composition can be administered by, for example, oral administration, subcutaneous injection, peritoneal administration, intravenous injection, intramuscular injection, paravertebral space (intradural) injection, sublingual administration, buccal administration, rectal insertion, vaginal injection, ocular administration, ear administration, nasal administration, inhalation, oral or nasal spray, skin administration, transdermal administration, etc.

[0131] Pharmaceutical composition of the present invention is determined by the type of medicine as active ingredient and various related factors (such as disease to be treated, route of administration, patient's age, sex and body weight and the severity of the disease). Specifically, the effective dose of composition according to the present invention can change according to patient's age, sex and body weight, and can usually be used every day or every other day or once to three times every day with every 1kg body weight 0.001 to 150mg composition, preferably every 1kg body weight uses 0.01 to 100mg composition. However, due to can increase or reduce preferred dose according to route of administration, the severity of the disease, sex, body weight, age etc., it is not intended to limit the scope of this preferred dose of the present invention by any method.

[0132] As used herein, "subject" refers to a subject in need of disease treatment, and more specifically refers to a mammal, such as a human or non-human primate, mouse, rat, dog, cat, horse, and cow.

[0133] As used herein, "administering" refers to providing a predetermined composition of the present invention to a subject in need thereof by any suitable method. As used herein, "prevention" refers to all actions that inhibit or delay the onset of a target disease, while "treatment" refers to all actions that improve or beneficially change the target disease and the metabolic abnormalities caused by it by administering a pharmaceutical composition according to the present invention, and "improvement" refers to all actions that alleviate the target disease and its related parameters (e.g., the severity of symptoms) by administering a composition according to the present invention.

[0134] The present invention provides a kit for preventing or treating graft-versus-host disease, autoimmune diseases or type II diabetes, and the kit comprises the carrier or the composition and instructions for use.

[0135] In addition to the above-mentioned components, the kit of the present invention may also include other components, devices and materials, etc., which are generally required for the expression method of the vector of the present invention and for storing, manipulating and enhancing the effect of the composition of the present invention. In addition, all components included in the kit can be used once or multiple times without any restrictions, there is no restriction on the order of use of each substance, and the application of each substance can be carried out simultaneously or in small increments.

[0136] In addition to the preparation and instructions for use, the test kit of the present invention may also include a container. The container may be used to package the configuration, and may also be used to store and fix it. The material used for the container may be in the form of, for example, a bottle, tub, sack, envelope, tube, and ampoule, which may be made in part or in whole of plastics, glass, paper, foil, and wax, etc. The container may be equipped with a wholly or partially detachable stopper, which may initially be a part of the container or may be attached to the container by machinery, adhesive, or other means, and may be equipped with a stopper that allows access to the contents by a syringe needle. The test kit may include an outer package, and the outer package may include instructions for use of the constituent elements, but is not limited thereto.

[0137] The following will provide preferred embodiments that are helpful for understanding the present invention. However, the following embodiments are provided only to make it easier to understand the present invention, and the content of the present invention is not limited to the following embodiments.

[0138] [Example]

[0139] Example 1. Discovery of anti-DR3 clones using phage display

[0140] Clones that specifically bind to DR3 were discovered by phage display. First, scFv clones that bind to DR3 were selectively amplified by three rounds of panning using a human scFv library and recombinant DR3. It was confirmed that at each stage, scFv clones that bind to recombinant DR3 were amplified by ELISA using phages obtained after panning. Thereafter, single clones confirmed by colony PCR were infected with phage, single clones that bind to DR3 were confirmed by ELISA, and the CDR region of each single clone was confirmed by sequencing.

[0141] The results showed that 5 clones that bind to DR3 ( Figure 1a and 1b ), and the five antibodies were named 2-10, 3-18, 4-4, 6-2 and 6-15. The five antibodies were sequenced, and the complementary determining region (CDR) of each clone was sequenced. Tables 1 and 2 show the amino acid and base sequences of CDR1 to CDR3 of the heavy chain region and variable region of each antibody. Table 3 shows the base sequence and amino acid sequence of the heavy chain region and variable region of each antibody. In each table, the bold text represents the CDR sequence according to IMGT, and the underlined portion represents the CDR sequence according to KABAT.

[0142] [Table 1]

[0143] SEQ ID NO: name sequence SEQ ID NO: 1 Heavy chain CDR1 of 2-10 mAb variable region <![CDATA[GYTF SYGIS ]]> SEQ ID NO: 2 Heavy chain CDR2 of 2-10 mAb variable region <![CDATA[ WISAYNGNTNYAQKLQG ]]> SEQ ID NO: 3 Heavy chain CDR3 of 2-10 mAb variable region <![CDATA[AS GPDYGMLDY ]]> SEQ ID NO: 4 Light chain CDR1 of 2-10 mAb variable region <![CDATA[ QASQDISN And]]> SEQ ID NO: 5 Light chain CDR2 of 2-10 mAb variable region <![CDATA[ DASNLET <!-- 13 -->]]> SEQ ID NO: 6 Light chain CDR3 of 2-10 mAb variable region <![CDATA[ QQYINLPLT ]]> SEQ ID NO: 7 Heavy chain CDR1 of 3-18 mAb variable region <![CDATA[GGSI SSSSYYWG ]]> SEQ ID NO: 8 Heavy chain CDR2 of 3-18 mAb variable region <![CDATA[ SIYYSGSTYYNPSLKS ]]> SEQ ID NO: 9 Heavy chain CDR3 of 3-18 mAb variable region <![CDATA[AB QVSGPLDY ]]> SEQ ID NO: 10 Light chain CDR1 of 3-18 mAb variable region <![CDATA[ RASQGIBNDLG ]]> SEQ ID NO: 11 Light chain CDR2 of 3-18 mAb variable region <![CDATA[ AASSLQS ]]> SEQ ID NO: 12 Light chain CDR3 of 3-18 mAb variable region <![CDATA[ LQDYNYPRT ]]> SEQ ID NO: 13 Heavy chain CDR1 of 4-4 mAb variable region <![CDATA[GFTFS DYYMS ]]> SEQ ID NO: 14 Heavy chain CDR2 of 4-4 mAb variable region <![CDATA[ YISSSSSYTNYADSVKG ]]> SEQ ID NO: 15 Heavy chain CDR3 of 4-4 mAb variable region <![CDATA[AS NVDTAMVGAFDI ]]> SEQ ID NO: 16 4-4 Light chain CDR1 of mAb variable region <![CDATA[ RASQHISNWLA ]]> SEQ ID NO: 17 4-4 Light chain CDR2 of mAb variable region <![CDATA[ TASNLQS ]]> SEQ ID NO: 18 4-4 Light chain CDR3 of mAb variable region <![CDATA[ QQADSFPLT ]]> SEQ ID NO: 19 Heavy chain CDR1 of 6-2 mAb variable region <![CDATA[GFTFS SYSMN ]]> SEQ ID NO: 20 Heavy chain CDR2 of 6-2 mAb variable region <![CDATA[ YISSSSSTIYYADSVKG ]]> SEQ ID NO: 21 Heavy chain CDR3 of 6-2 mAb variable region <![CDATA[AB WDGYCSGGSCYSGMDY ]]> SEQ ID NO: 22 Light chain CDR1 of 6-2 mAb variable region <![CDATA[ QASQDISNYLN ]]> SEQ ID NO: 23 Light chain CDR2 of 6-2 mAb variable region <![CDATA[ DASNLET ]]> SEQ ID NO: 24 Light chain CDR3 of 6-2 mAb variable region <![CDATA[ QQYINLPLT ]]> SEQ ID NO: 25 Heavy chain CDR1 of 6-15 mAb variable region <![CDATA[GFTFS SYSMN ]]> SEQ ID NO: 26 Heavy chain CDR2 of 6-15 mAb variable region <![CDATA[ YISSSSSTIYYADSVKG ]]> SEQ ID NO: 27 Heavy chain CDR3 of 6-15 mAb variable region <![CDATA[AB WDGYCSGGSCYSGMDY ]]> SEQ ID NO: 28 Light chain CDR1 of 6-15 mAb variable region <![CDATA[ QASQDISNYLN ]]> SEQ ID NO: 29 Light chain CDR2 of 6-15 mAb variable region <![CDATA[ DASNLET ]]> SEQ ID NO: 30 Light chain CDR3 of 6-15 mAb variable region <![CDATA[ QQYINLPLT ]]>

[0144] [Table 2]

[0145]

[0146] [Table 3]

[0147]

[0148]

[0149]

[0150] Example 2. Confirmation of the excellent DR3-specific binding strength of five antibodies

[0151] The specific binding strength of the five antibodies developed in Example 1 to DR3 was confirmed by ELISA, SPR and FACS experiments.

[0152] Example 2-1. Confirmation of specific binding strength to DR3 by ELISA

[0153] ELISA was performed using monoclonal antibodies from the clones selected in Example 1. Specifically, 0.4 μg / ml DR3-His tag was added to PBS and mixed with PBS, and then the Ni plate was coated with a total of 50 μl of the mixed solution. Thereafter, in order to prevent nonspecific binding, the Ni plate was treated with 200 μl of PBS containing 5% FBS and incubated at room temperature for 30 minutes, and incubated for 30 minutes by titration of the concentration of the monoclonal antibody. The degree of binding between the coated DR3 and the monoclonal antibody was detected with anti-human IgG Fc HRP. The plate was washed five times with 250 μl 0.1% Tween 20 at each step. The isotype control was used as a negative control.

[0154] The results confirmed that the five antibodies of the present invention had excellent specific binding ability to DR3 compared with the control, among which 4-4, 6-2 and 6-15 maintained significantly higher binding specificity to DR3 even at low antibody concentrations ( Figure 2a ).

[0155] Example 2-2. Confirmation of specific binding strength to DR3 by SPR

[0156] SPR was performed using monoclonal antibodies from the clones selected in Example 1. After immobilizing 10 μg / ml DR3-His tag on a CM5 chip, binding specificity and binding strength were measured using Biacore T100 under conditions of a contact time of 30 seconds and a flow rate of 30 μl / min.

[0157] As a result, it was found that the five antibodies according to Example 1 had significantly lower DR3 binding dissociation constants compared to the DR3 agonist TL1A, and were confirmed to exhibit binding strengths of about 12 to 9,200 times or more ( Figure 2b ).

[0158] Example 2-3. Confirmation of specific binding strength to DR3 by FACS

[0159] FACS was performed using monoclonal antibodies from the clones selected in Example 1. Specifically, a concentration of 0.2×10 6 The DR3 stable cell line was mixed with 2 μg / ml of monoclonal antibody and incubated at 4°C for 30 minutes, and the binding degree between the DR3 stable cell line and the monoclonal antibody was confirmed by FACS analysis using anti-human IgG PE. The cells were washed once with 200 μl PBS at each step. The isotype control was used as a negative control.

[0160] The results confirmed that antibodies 4-4, 6-2, and 6-15 strongly bound to DR3 ( Figure 2c ).

[0161] Example 3. Epitope identification of five antibodies against DR3

[0162] Example 3-1. Epitope identification by ELISA

[0163] ELISA was performed to confirm whether the epitope of rDR3 bound by the anti-DR3 4-4 antibody among the five antibodies in Example 1 overlapped with the ligand TL1A of rDR3 or with other monoclonal antibodies. Specifically, 0.4 μg / ml DR3-His tag was added to PBS and mixed with PBS, and then the Ni plate was coated with a total of 50 μl of the mixed solution. Thereafter, in order to prevent nonspecific binding, the Ni plate was mixed with 200 μl of PBS containing 5% FBS and incubated at room temperature for 30 minutes, and incubated for 30 minutes by titrating the concentrations of TL1A and monoclonal antibodies. Thereafter, without washing, 200 ng / ml of biotinylated anti-DR3 4-4 antibody was incubated for 30 minutes, and anti-human IgG Fc HRP was used to detect the degree of binding between DR3 and biotinylated anti-DR3 4-4 antibody. Except for the above steps without washing, the plate was washed 5 times with 250 μl of 0.1% Tween20 in each step. Isotype controls were used as negative controls.

[0164] Example 3-2. Epitope identification by FACS

[0165] FACS was performed to confirm whether the epitope of DR3 expressed on the cell surface that bound to the five selected antibodies overlapped with the ligand TL1A of human DR3 or with other monoclonal antibodies. 6 The DR3 stable cell line was mixed with 2 μg / ml monoclonal antibody and incubated at 4°C for 30 minutes, and then without washing, it was mixed with 0.625 μg / ml TL1A His tag, and the resulting mixture was incubated at 4°C for 30 minutes. The degree of binding between the DR3 stable cell line and the TL1A-His tag was detected using anti-His tag AF647 and confirmed by FACS. Except for the above steps without washing, the cells were washed once with 200 μl PBS in each step. The isotype control was used as a negative control.

[0166] The results confirmed that clones 4-4, 6-2, and 6-15 bind to the epitope that binds to DR3 and TL1A, whereas clones 2-10 and 3-18 do not bind to the epitope ( Figure 3a and 3b ).

[0167] Example 4. Confirmation of the excellent NF-κB activation effect of the selected antibodies

[0168] In order to confirm the NF-κB activation effect of the antibodies selected in Example 1, the phosphorylated NF-κB (pNF-κB) level was compared and analyzed by FACS. Specifically, clone 4-4 IgG1, clone 2-10 IgG1, clone 3-18 IgG1, and clone 6-2 IgG1 monoclonal antibodies and rhTL1A were used to activate pNF-κB. 1×10 6 hPBMCs were mixed with rhTL1A-His tag or titrated monoclonal antibodies and then incubated for 10 minutes at 37° C. The activation level of NF-κB (pNF-κB) was analyzed using FACS by anti-human phospho-NF-κB p65 (Ser529) eFlour660.

[0169] The results confirmed that 4-4 (IgG1) had the highest level of NF-κB activation, followed by 6-2. This was found to be because the site where the antibody binds to DR3 overlaps with TL1A and strongly phosphorylates NF-κB ( Figure 4 ).

[0170] Example 5. Comparison of binding strength according to IgG subclass of clone 4-4

[0171] The functional best subclass of the anti-DR3 4-4 antibody identified as the best in Example 2 was analyzed. To this end, clone 4-4IgG1, clone 4-4IgG2 and clone 4-4IgG4 monoclonal antibodies were prepared as anti-DR3 4-4 antibodies according to IgG subclass, and ELISA was performed to compare their binding affinity to DR3. Specifically, 0.4 μg / ml DR3-His tag was added to PBS and mixed with PBS, and then Ni plates were coated with a total of 50 μl of the mixed solution. Thereafter, in order to prevent nonspecific binding, the Ni plate was mixed with 200 μl of PBS containing 5% FBS and incubated at room temperature for 30 minutes, and incubated for 30 minutes by monoclonal antibody titration. The degree of binding between the coated DR3 and the monoclonal antibody was detected with anti-human IgG Fc HRP. The plate was washed five times with 250 μl of 0.1% Tween 20 at each step.

[0172] The results confirmed that the anti-DR3 4-4 antibody had excellent affinity for DR3 in the order of IgG1, IgG4, and IgG2 subclasses ( Figure 5 ).

[0173] Example 6. Confirmation of excellent NF-κB activation effect based on IgG subclass of clone 4-4

[0174] In order to compare the NF-κB activation levels according to the IgG subclass of clone 4-4, FACS was performed after stimulation with clone 4-4IgG1, clone 4-4IgG2, and clone 4-4IgG4 monoclonal antibodies. 6 hPBMCs were mixed with rhTL1A-His tag or titrated monoclonal antibodies and then incubated for 10 minutes at 37° C. NF-κB activation levels were analyzed using FACS with anti-human phospho-NF-κB p65 (Ser529) eFlour 660.

[0175] The results confirmed that antibody 4-4 had a high level of NF-κB activation in the order of IgG4, IgG1, and IgG2 subclasses ( Figure 6 ).

[0176] Example 7. Effect of anti-DR3 4-4 IgG4 antibody on the survival rate of regulatory T cells in the absence of TCR signal stimulation

[0177] In order to confirm whether the anti-DR3 4-4 IgG4 antibody confirmed as the best in Example 6 can improve the survival rate of human regulatory T cells without TCR signal stimulation, CD4 T cell culture was performed using the anti-DR3 4-4 IgG4 antibody. Specifically, after CD4 T cells were isolated from hPBMC using a human CD4 T cell enrichment kit, the cells were cultured with 20 ng / ml IL-2, 15 ng / ml TL1A or 1 μg / ml anti-DR3 4-4 IgG4 antibody for 3 days and then analyzed by FACS.

[0178] The results confirmed that in the absence of TCR signaling, anti-DR3 4-4 IgG4 antibodies could increase the survival of regulatory T cells to a level similar to that caused by the DR3 ligand TL1A ( Figure 7 ).

[0179] Example 8. Effect of anti-DR3 4-4 IgG4 antibody on the division and survival rate of regulatory T cells according to TCR signal strength

[0180] In order to confirm the degree of promotion of human regulatory T cell division by the anti-DR3 4-4IgG4 antibody confirmed as the best in Example 6 according to the intensity of TCR signal stimulation, CD4T cell culture was performed using anti-hCD3 antibody and anti-DR3 4-4IgG4 antibody. Specifically, one day before cell culture, 96-well cell culture plates were coated with 1.5μg / ml, 0.5μg / ml, 0.25μg / ml, 0.125μg / ml and 0μg / ml of anti-hCD3ε antibody, respectively. Then, on the day of cell culture, CD4 T cells were isolated from hPBMC using a human CD4T cell enrichment kit. Thereafter, 0.1x10 6 CD4 T cells were cultured for 6 days by adding 20 ng / ml IL-2, 0.5 ng / ml TGF-β, or anti-DR3 4-4 IgG4 antibody according to each group, and then analyzed by FACS.

[0181] As a result, it was confirmed that when a certain intensity or higher TCR signal stimulation was given, the anti-DR3 4-4 IgG4 antibody effectively increased the division or survival rate of regulatory T cells ( Figure 8 ).

[0182] Example 9. Sequencing of anti-DR3 4-4 IgG4 monoclonal antibody

[0183] The anti-DR3 4-4 IgG4 monoclonal antibody identified as the most functional in Examples 3 to 8 was sequenced for application in the treatment of graft-versus-host disease, autoimmune diseases and type II diabetes. Specifically, based on the sequence analysis data, the Fab region of 4-4 mAb was cloned into an IgG4 expression vector, and the DNA sequence and amino acid sequence of the cloned vector are shown in Table 4. Table 4 shows the DNA sequence and amino acid sequence of the complete human 4-4.

[0184] [Table 4]

[0185]

[0186] The above description of the present invention is provided for illustrative purposes, and those skilled in the art will understand that the present invention can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. Therefore, it should be understood that the above embodiments in all aspects are only illustrative and not restrictive.

[0187] [Industrial Applicability]

[0188] The antibody that specifically binds to human DR3 can specifically bind to the epitope to which the TLA1 ligand of human DR3 binds, and can selectively act on regulatory T cells to minimize the activity of effector CD4+T cells and CD8+T cells while increasing the division or survival rate of regulatory T cells, so the antibody can be effectively used as a therapeutic agent for graft-versus-host disease, autoimmune diseases or type II diabetes, and is therefore considered to be industrially useful.

Claims

1. An anti-death receptor 3 (DR3) antibody or an antigen-binding fragment thereof, wherein the anti-DR3 antibody or the antigen-binding fragment thereof specifically binds to DR3.

2. The anti-DR3 antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof is any one selected from the group consisting of: i) an antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a heavy chain CDR 1 as shown in the amino acid sequence of SEQ ID NO: 1, a heavy chain CDR 2 as shown in the amino acid sequence of SEQ ID NO: 2, and a heavy chain CDR 3 as shown in the amino acid sequence of SEQ ID NO: 3; and the light chain variable region comprises a light chain CDR 1 as shown in the amino acid sequence of SEQ ID NO: 4, a light chain CDR 2 as shown in the amino acid sequence of SEQ ID NO: 5, and a light chain CDR 3 as shown in the amino acid sequence of SEQ ID NO: 6; ii) an antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the heavy chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 7, the heavy chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 8, and the heavy chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 9; and the light chain variable region comprises the light chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 10, the light chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 11, and the light chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 12; iii) an antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a heavy chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 13, a heavy chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 14, and a heavy chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 15; and the light chain variable region comprises a light chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 16, a light chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 17, and a light chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 18; iv) an antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a heavy chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 19, a heavy chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 20, and a heavy chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 21; and the light chain variable region comprises a light chain CDR 1 set forth in the amino acid sequence of SEQ ID NO: 22, a light chain CDR 2 set forth in the amino acid sequence of SEQ ID NO: 23, and a light chain CDR 3 set forth in the amino acid sequence of SEQ ID NO: 24; and v) an antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the heavy chain CDR 1 shown in the amino acid sequence of SEQ ID NO: 25, the heavy chain CDR 2 shown in the amino acid sequence of SEQ ID NO: 26, and the heavy chain CDR 3 shown in the amino acid sequence of SEQ ID NO: 27; and the light chain variable region comprises the light chain CDR 1 shown in the amino acid sequence of SEQ ID NO: 28, the light chain CDR 2 shown in the amino acid sequence of SEQ ID NO: 29, and the light chain CDR 3 shown in the amino acid sequence of SEQ ID NO:

30.

3. The anti-DR3 antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof is any one selected from the group consisting of: i) an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 63 and a light chain variable region represented by the amino acid sequence of SEQ ID NO: 64; ii) an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 67 and a light chain variable region represented by the amino acid sequence of SEQ ID NO: 68; iii) an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 71 and a light chain variable region represented by the amino acid sequence of SEQ ID NO: 72; iv) an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 75 and a light chain variable region represented by the amino acid sequence of SEQ ID NO: 76; and v) an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 79 and a light chain variable region represented by the amino acid sequence of SEQ ID NO:

80.

4. The anti-DR3 antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain represented by the amino acid sequence of SEQ ID NO: 83 and a light chain represented by the amino acid sequence of SEQ ID NO:

84.

5. The anti-DR3 antibody or antigen-binding fragment thereof according to claim 4, wherein the antibody or antigen-binding fragment comprises Fab. The anti-DR3 antibody or antigen-binding fragment thereof according to claim 1 , wherein the antibody or antigen-binding fragment is of IgG1 subclass or IgG4 subclass.

7. The anti-DR3 antibody or antigen-binding fragment thereof according to claim 1, wherein the antigen-binding fragment is any one selected from the group consisting of Fv, Fab, F(ab')2, Fab', dsFv, scFv, sc(Fv)2 and diabody.

8. The anti-DR3 antibody or antigen-binding fragment thereof according to claim 2, wherein the antibody or antigen-binding fragment of iii), iv) and v) specifically binds to DR3 in competition with TL1A.

9. The anti-DR3 antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof is any one or more of the following: Selectively activate regulatory T cells or enhance their survival; and Activates NF-κB.

10. A nucleic acid sequence encoding the antibody or antigen-binding fragment according to claim 1.

11. A vector comprising the nucleic acid sequence according to claim 10.

12. A transgenic organism transformed with the vector according to claim 11.

13. A pharmaceutical composition for preventing or treating graft-versus-host disease, autoimmune diseases or type II diabetes, comprising the anti-DR3 antibody or antigen-binding fragment thereof according to claim 1 that specifically binds to death receptor 3 (DR3) as an active ingredient. 14 . The pharmaceutical composition according to claim 13 , wherein the antibody or antigen-binding fragment thereof comprises a heavy chain represented by the amino acid sequence of SEQ ID NO: 83 and a light chain represented by the amino acid sequence of SEQ ID NO:

84.

15. The pharmaceutical composition according to claim 13, wherein the autoimmune disease is any one or more selected from the group consisting of: an autoimmune disease caused by graft-versus-host disease, hemophagocytic lymphohistiocytosis, systemic lupus erythematosus, Kikuchi's disease, adult-onset Still's disease, Behcet's disease, IgG4-related disease, type I diabetes, systemic sclerosis, psoriasis, multiple sclerosis and Graves' hyperthyroidism.

16. A kit for preventing or treating graft-versus-host disease, autoimmune diseases or type II diabetes, the kit comprising instructions for use and the vector according to claim 11 or the composition according to claim 13.

17. A method for preventing or treating graft-versus-host disease, autoimmune diseases or type II diabetes, the method comprising administering to a subject in need thereof a composition comprising an anti-DR3 antibody or an antigen-binding fragment thereof, wherein the anti-DR3 antibody or the antigen-binding fragment thereof binds to death receptor 3 (DR3) as an active ingredient.

18. Use of a composition comprising an anti-DR3 antibody or an antigen-binding fragment thereof as an active ingredient that binds to death receptor 3 (DR3) for preventing or treating graft-versus-host disease, autoimmune diseases or type II diabetes.

19. Use of a composition comprising an anti-DR3 antibody or an antigen-binding fragment thereof as an active ingredient that binds to death receptor 3 (DR3) in the preparation of a preparation for preventing or treating graft-versus-host disease, autoimmune diseases or type II diabetes.

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