Antibodies that specifically bind to human DR3 and their applications

Abstract

The present invention provides an anti-DR3 antibody or its antigen-binding fragment that specifically binds to the cell death receptor 3 (DR3). [Solution] The present invention relates to an antibody that specifically binds to human DR3 and its applications, which specifically forms a binding to a specific epitope of DR3 and selectively acts on regulatory T cells to exhibit effector activity on CD4 + T cells and CD8 + This method can minimize T cell activity while strongly activating only regulatory T cells. By inducing such selective regulatory T cell activity, it exhibits remarkably superior preventive or therapeutic effects against graft-versus-host disease, autoimmune diseases, and type 2 diabetes, making it useful as a therapeutic agent for a variety of related diseases.

Description

[Technical Field]

[0001] This invention relates to an antibody that specifically binds to human DR3 and its applications.

[0002] This application is the Republic of Korea Patent Application No. 10-2022-0, filed on June 17, 2022. Patent No. 074078 and Republic of Korea Patent Application No. 10-20 filed on June 15, 2023 Claiming the benefit of priority under No. 23-0077023, and the specification and drawings of said application All information presented herein is incorporated herein by reference. [Background technology]

[0003] Death receptor 3 (DR3, tumor necro) sis factor receptor superfamily 25, TNFRS F25) is one of the TNFRSF members. DR3 consists of 417 amino acids. It is composed of 47 kDa, and its molecular weight is derived from the homotrimer. DR3 is expressed in large quantities from regulatory T cells (Tregs) in the steady state, but Conventional T cells also increase in expression when activated. In addition, DR3 is a type of nematode cell, which includes NK cells, ILCs, B cells, mononuclear cells, and non- -It is known to be expressed in plague cells such as osteocytes and endothelial cell colony-forming cells.

[0004] TL1A (tumor necrosis factor-ligand) is a ligand for DR3. ke ligand 1A) is activated by T cells, dendritic cells, macrophages, mononuclear cells, and other cells. It is primarily expressed in cortical cells. TL1A activates NF-κB signaling by binding to DR3. This results in T cells receiving costimulation factors. It functions as such. According to existing reports, DR3 agonist This can be used to induce selective activation of regulatory T cells. At this time, DR3 jaw One method for activating regulatory T cells is to use effervescent cells outside of regulatory T cells. Unlike IL-2, which also activates T cells, it selectively activates only regulatory T cells. Activation of effector T cells is the main factor that interferes with treatment when treating various inflammatory diseases. This could become a problem.

[0005] DR3 agonists are TL1A, which is a ligand for DR3, or agonistic anti-DR3. 3 antibodies may also be used. However, TL1A is a natural inhibitor. DcR3 (decoy receptor), which is a bitor, exists, and cytokines Since the natural inhibitors are usually regulated by negative feedback, using TL1A Methods for inducing DR3 activation are difficult for efficient drug delivery, and homotrimer cytocytokines Due to this characteristic, it can increase the difficulty of the new drug development process. On the other hand, agonistic antibodies - Using a DR3 antibody avoids negative feedback by DcR3 and enables efficient DR 3. Activation can be induced.

[0006] On the other hand, antibody drugs were first developed in 1975 with the introduction of monoclonal antibodies. Since the development of the tibody, research into its clinical applicability has been continuously conducted. It exhibits strong binding affinity and high binding specificity towards the antigen. By utilizing the properties of antibodies with (binding specificity), specific organisms The mechanism of action that can strongly and selectively control the learned reaction is the fundamental core of the development potential of antibody drugs. In particular, compared with existing therapeutic agents based on chemically synthesized substances, anti- body drugs have relatively few side effects due to their high binding specificity and stability in the human body, and their therapeutic efficacy is excellent. Therefore, the field of developing therapeutic agents using antibodies has attracted much attention as the next-generation core field of new drug research and development.

[0007] However, an antibody drug that specifically binds to DR3, activates regulatory T cells, and has excellent therapeutic effects on graft-versus-host disease, autoimmune diseases or type 2 diabetes has not been developed so far.

Summary of the Invention

Problems to be Solved by the Invention

[0008] One object of the present invention is to provide an anti-DR3 antibody or an antigen-binding fragment thereof that specifically binds to death receptor 3 (DR3).

[0009] Another object of the present invention is to provide a nucleic acid sequence encoding the antibody or antigen-binding fragment.

[0010] Another object of the present invention is to provide a vector containing the nucleic acid sequence.

[0011] Another object of the present invention is to provide a transformant transformed with the vector.

[0012] Another object of the present invention is to provide a transplant containing an anti-DR3 antibody or an antigen-binding fragment thereof that specifically binds to death receptor 3 (DR3) as an active ingredient. ​​​​​​To provide a pharmaceutical composition for preventing or treating graft-versus-host disease, autoimmune diseases or type 2 diabetes is what is needed.

[0013] Another object of the present invention is to provide a kit for preventing or treating graft-versus-host disease, autoimmune diseases or type 2 diabetes, including the vector or the composition, and a manual. is what is needed.

[0014] However, the technical problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those with ordinary knowledge in the technical field to which the present invention belongs from the following description. should be clearly understood by those with ordinary knowledge in the technical field to which the present invention belongs from the following description.

Means for Solving the Problems

[0015] The present invention provides an anti-DR3 antibody that specifically binds to death receptor 3 (DR3) or an antigen-binding fragment thereof. is what is needed.

[0016] The present invention provides a nucleic acid sequence encoding the antibody or antigen-binding fragment.

[0017] The present invention provides a vector containing the nucleic acid sequence.

[0018] The present invention provides a transformant transformed with the vector.

[0019] The present invention provides a composition for preventing or treating graft-versus-host disease, autoimmune diseases or type 2 diabetes, containing an anti-DR3 antibody that specifically binds to death receptor 3 (DR3) or an antigen-binding fragment thereof as an active ingredient. is what is needed. is what is needed.

[0020] The present invention provides a kit for preventing or treating graft-versus-host disease, autoimmune diseases or type 2 diabetes, including the vector or the composition, and a manual. To provide a kit for the prevention or treatment of infectious diseases or type 2 diabetes.

[0021] In one embodiment of the present invention, the antibody or the antigen-binding fragment is selected from the group consisting of the following. It may be any one of the following, but is not limited to:

[0022] i) The amino acid sequence of the heavy chain CDR1 described in SEQ ID NO: 1, and the amino acid sequence of SEQ ID NO: 2 Includes heavy chain CDR2 as described and heavy chain CDR3 as described in the amino acid sequence of SEQ ID NO: 3. Heavy chain variable region; and light chain CDR1, described by the amino acid sequence of SEQ ID NO: 4, SEQ ID NO: 5 Light chain CDR2 and light chain SEQ ID NO: 6 described by amino acid sequence Antibodies or antigen-binding fragments containing a light chain variable region including CDR3; ii) The amino acid sequence of heavy chain CDR1 described in the amino acid sequence of SEQ ID NO: 7, and the amino acid sequence of SEQ ID NO: 8 It contains heavy chain CDR2 as described in and heavy chain CDR3 as described in the amino acid sequence of SEQ ID NO: 9. The heavy chain variable region; and the light chain CDR1, described by the amino acid sequence of SEQ ID NO: 10, SEQ ID NO: The light chain CDR2, described by the amino acid sequence of 11 amino acids, and the amino acid sequence of SEQ ID NO: 12 An antibody or antigen-binding fragment containing a light chain variable region including a light chain CDR3; iii) The heavy chain CDR1 described in the amino acid sequence of SEQ ID NO: 13, and the amino acid sequence of SEQ ID NO: 14 Heavy chain CDR2 described by its acid sequence and heavy chain CD described by the amino acid sequence of SEQ ID NO: 15 Heavy chain variable region including R3; and light chain CDR1 as described by the amino acid sequence of SEQ ID NO: 16, The amino acid sequence of the light chain CDR2 described in SEQ ID NO: 17 and the amino acid sequence of SEQ ID NO: 18 An antibody or antigen-binding fragment containing a light chain variable region including the light chain CDR3 as described in [reference]; iv) The amino acids of heavy chain CDR1 described in the amino acid sequence of SEQ ID NO: 19, and the amino acids of SEQ ID NO: 20 Heavy chain CDR2 described by sequence and heavy chain CDR described by amino acid sequence of SEQ ID NO: 21 A heavy chain variable region containing 3; and the light chain CDR1 described in the amino acid sequence of SEQ ID NO: 22, The amino acid sequence of the light chain CDR2 described in the amino acid sequence of column number 23 and the amino acid sequence of SEQ ID NO: 24 Antibodies or antigen-binding fragments containing a light chain variable region including the described light chain CDR3; and v) The amino acid sequence of heavy chain CDR1 described in SEQ ID NO: 25, and the amino acid sequence of SEQ ID NO: 26 Heavy chain CDR2 described in columns and heavy chain CDR3 described in the amino acid sequence of SEQ ID NO: 27 Heavy chain variable region including; and light chain CDR1, sequence described in the amino acid sequence of SEQ ID NO: 28. The amino acid sequence of the light chain CDR2, described as amino acid sequence number 29, and the amino acid sequence of sequence number 30 are described. An antibody or antigen-binding fragment containing a light chain variable region including the listed light chain CDR3.

[0023] In one embodiment of the present invention, the antibody or the antigen-binding fragment is selected from the group consisting of the following. It may be any one of the following, but is not limited to: i) The heavy chain variable region described in the amino acid sequence of SEQ ID NO: 63 and the amino acids of SEQ ID NO: 64 Antibodies or antigen-binding fragments containing a light chain variable region described by sequence; ii) The heavy chain variable region described in the amino acid sequence of SEQ ID NO: 67 and the amino acid sequence of SEQ ID NO: 68 Antibodies or antigen-binding fragments containing a light chain variable region described by an acid sequence; iii) The heavy chain variable region described in the amino acid sequence of SEQ ID NO: 71 and the amino acid sequence of SEQ ID NO: 72 Antibodies or antigen-binding fragments containing a light chain variable region described by an acid sequence; iv) The heavy chain variable region described in the amino acid sequence of SEQ ID NO: 75 and the amino acid sequence of SEQ ID NO: 76 Antibodies or antigen-binding fragments containing light chain variable regions described by acid sequences; and v) The heavy chain variable region described in the amino acid sequence of SEQ ID NO: 79 and the amino acids of SEQ ID NO: 80 An antibody or antigen-binding fragment containing a light chain variable region described by sequence.

[0024] In one embodiment of the present invention, the antibody or the antigen-binding fragment is the amino acid of SEQ ID NO: 83. It may contain the heavy chain described in the sequence and the light chain described in the amino acid sequence of SEQ ID NO: 84. It is possible, but it is not limited to this.

[0025] In one embodiment of the present invention, the antibody or the antigen-binding fragment may include Fab. However, it is not limited to this.

[0026] In one embodiment of the present invention, the antibody or the antigen-binding fragment is IgG1 or IgG4 It may be a subclass, but is not limited to that.

[0027] In one embodiment of the present invention, the antigen-binding fragment is Fv, Fab, F(ab')2, Fa From b', dsFv, scFv, sc(Fv)2 and diabody You may choose any one of the following groups, but you are not limited to this.

[0028] In one embodiment of the present invention, the antibodies or antigen-binding fragments described in iii), iv) and v) are T While L1A can compete with DR3 for specific binding, it is not limited to this.

[0029] In one embodiment of the present invention, the antibody or the antigen-binding fragment is one of the following: The above are possible characteristics, but are not limited to them: Selectively activate regulatory T cells or increase their survival rate; and Activates NF-κB.

[0030] In one embodiment of the present invention, the autoimmune disease is an autoimmune disease caused by graft-versus-host disease. Hemophagocytic lymphohistiocytosis cytosis, systemic lupus erythematosus hematosus, Kikuchi disease, adult-onset Still's disease Behcet's disease, IgG4 -Related diseases, type 1 diabetes, systemic sclerosis , psoriasis, multiple sclerosis, and Graves' thyroid Selected from a group consisting of Graves hyperthyroidism You may include one or more of these, but you are not limited to this.

[0031] Furthermore, anti-D2 binds to death receptor 3 (DR3). R3 antibody or its antigen-binding fragment, or a composition containing the same as an active ingredient, is required. The prevention of graft-versus-host disease, autoimmune disease, or type 2 diabetes, including the step of administering it to the individual. We provide treatment methods.

[0032] Furthermore, anti-D2 binds to death receptor 3 (DR3). Graft-versus-host disease: R3 antibody or its antigen-binding fragment, or a composition containing it as an active ingredient. It provides applications for the prevention or treatment of autoimmune diseases or type 2 diabetes.

[0033] Furthermore, anti-D2 binds to death receptor 3 (DR3). Graft-versus-host disease: R3 antibody or its antigen-binding fragment, or a composition containing it as an active ingredient. It provides applications for manufacturing preventive or therapeutic agents for autoimmune diseases or type 2 diabetes. [Effects of the Invention]

[0034] Antibodies that specifically bind to human DR3 and their applications are specific to the particular epitope of human DR3. It binds precisely to regulatory T cells and selectively acts on effector CD4+ T cells and CD8+ T cells. This can minimize T cell activity while improving the division or survival rate of regulatory T cells. In terms of its remarkable effectiveness in preventing or treating related diseases due to such activity, the graft It can be usefully utilized as a therapeutic agent for host-versus-host diseases, autoimmune diseases, or type 2 diabetes. [Brief explanation of the drawing]

[0035] [Figure 1] Figures 1(a) and 1(b) show experimental results of the process of discovering anti-DR3 scFv clones that specifically bind to DR3 via phage display. [Figure 2] Figures 2(a) to 2(c) show the binding specificity and affinity of anti-DR3 monoclonal antibodies to DR3. [Figure 3] Figures 3(a) and 3(b) show the results confirming that the binding site of the anti-DR3 4-4 antibody against DR3 is superimposed on the binding site of TL1A. [Figure 4] Figure 4 shows the NF-κB activation level of the anti-DR3 antibody of the present invention. [Figure 5] Figure 5 shows the results of comparing the affinity of anti-DR3 4-4 antibodies to DR3 based on IgG subclass. [Figure 6] Figure 6 shows the results of a comparative analysis of NF-κB activation levels of anti-DR3 4-4 antibodies based on IgG subclasses. [Figure 7] Figure 7 shows the effect of anti-DR34-4IgG4 antibody on increasing the survival rate of regulatory T cells in the absence of TCR signaling. [Figure 8] Figure 8 shows the effect of anti-DR3 4-4 IgG4 antibody on increasing the division and survival rate of regulatory T cells, as measured by TCR signal intensity. [Modes for carrying out the invention]

[0036] In this invention, fully human phage display and DR3 expression are achieved. Binding to DR3 natural form using stable cell lines We discovered an anti-DR3 antibody that reacts to the DR3 expressed by human T cells. The anti-DR3 antibody of the present invention reacts to the DR3 expressed by human T cells. Activating T cells through competitive binding with TL1A to 3, thereby acting as an agonist. We confirmed that this is possible. Furthermore, clone 4, which most strongly activates T cells, was identified. -4 has different isotypes from each other, such as IgG1, IgG2, and IgG4. Antibodies were produced and the degree of T cell activation was compared according to the antibody isotype, and anti-DR3 IgG4 antibodies specifically bind to DR3 natural form and most strongly target T cells. We confirmed that it activates. Furthermore, through this, we confirmed that it is the most superior anti -DR3 4-4 IgG4 antibody increases regulatory T cell viability in the absence of TCR stimulation. Therefore, when there is TCR signal stimulation of a certain intensity or higher, the division or survival rate of regulatory T cells is effectively increased. The present invention was completed by confirming that it could be increased to [a certain value].

[0037] This invention specifically binds to the cell death receptor 3 (DR3). The present invention provides an anti-DR3 antibody or its antigen-binding fragment that can be combined with it.

[0038] In this invention, DR3 (TNFRSF25, TNF receptor superfamily member 25, The cytoplasmic death receptor 3) is one of the tumor death factor receptor superfamily, and is regulatory Although it is expressed at high levels from T cells, it can also be induced from existing T cells upon TCR stimulation. In this invention, DR3 may include, but is not limited to, DR3, hDR3, etc. No. DR3 signaling has been validated in mice using agonist anti-DR3 antibodies. Furthermore, it can promote the activation and proliferation of regulatory T cells under normal conditions, and is non-targeted even in small amounts. It is possible to induce the proliferation of regulatory T cells without the proliferation of immune cell populations, and this is CD4 and This is a major difference from the IL-2 complex, which also promotes the division of CD8 T cells. The division of conventional T cells is involved in the treatment of various inflammatory diseases. Because this can be a major obstacle to treatment, anti-DR3 antibodies selectively activate regulatory T cells. It can be used as a new method.

[0039] In one embodiment of the present invention, the antibody or the antigen-binding fragment is selected from the group consisting of the following. There may be one of these, but it is not limited to this: i) The amino acid sequence of the heavy chain CDR1 described in SEQ ID NO: 1, and the amino acid sequence of SEQ ID NO: 2 Includes heavy chain CDR2 as described and heavy chain CDR3 as described in the amino acid sequence of SEQ ID NO: 3. Heavy chain variable region; and light chain CDR1, described by the amino acid sequence of SEQ ID NO: 4, SEQ ID NO: 5 Light chain CDR2 and light chain SEQ ID NO: 6 described by amino acid sequence Antibodies or antigen-binding fragments containing a light chain variable region including CDR3; ii) The amino acid sequence of heavy chain CDR1 described in the amino acid sequence of SEQ ID NO: 7, and the amino acid sequence of SEQ ID NO: 8 It contains heavy chain CDR2 as described in and heavy chain CDR3 as described in the amino acid sequence of SEQ ID NO: 9. The heavy chain variable region; and the light chain CDR1, described by the amino acid sequence of SEQ ID NO: 10, SEQ ID NO: The light chain CDR2, described by the amino acid sequence of 11 amino acids, and the amino acid sequence of SEQ ID NO: 12 An antibody or antigen-binding fragment containing a light chain variable region including a light chain CDR3; iii) The heavy chain CDR1 described in the amino acid sequence of SEQ ID NO: 13, and the amino acid sequence of SEQ ID NO: 14 Heavy chain CDR2 described by its acid sequence and heavy chain CD described by the amino acid sequence of SEQ ID NO: 15 Heavy chain variable region including R3; and light chain CDR1 as described by the amino acid sequence of SEQ ID NO: 16, The amino acid sequence of the light chain CDR2 described in SEQ ID NO: 17 and the amino acid sequence of SEQ ID NO: 18 An antibody or antigen-binding fragment containing a light chain variable region including the light chain CDR3 as described in [reference]; iv) The amino acids of heavy chain CDR1 described in the amino acid sequence of SEQ ID NO: 19, and the amino acids of SEQ ID NO: 20 Heavy chain CDR2 described by sequence and heavy chain CDR described by amino acid sequence of SEQ ID NO: 21 A heavy chain variable region containing 3; and the light chain CDR1 described in the amino acid sequence of SEQ ID NO: 22, The amino acid sequence of the light chain CDR2 described in the amino acid sequence of column number 23 and the amino acid sequence of SEQ ID NO: 24 Antibodies or antigen-binding fragments containing a light chain variable region including the described light chain CDR3; and v) The amino acid sequence of heavy chain CDR1 described in SEQ ID NO: 25, and the amino acid sequence of SEQ ID NO: 26 Heavy chain CDR2 described in columns and heavy chain CDR3 described in the amino acid sequence of SEQ ID NO: 27 Heavy chain variable region including; and light chain CDR1, sequence described in the amino acid sequence of SEQ ID NO: 28. The amino acid sequence of the light chain CDR2, described as amino acid sequence number 29, and the amino acid sequence of sequence number 30 are described. An antibody or antigen-binding fragment containing a light chain variable region including the listed light chain CDR3.

[0040] In one embodiment of the present invention, the antibody or the antigen-binding fragment is selected from the group consisting of the following. It may be any one of the following, but is not limited to: i) The heavy chain variable region described in the amino acid sequence of SEQ ID NO: 63 and the amino acids of SEQ ID NO: 64 Antibodies or antigen-binding fragments containing a light chain variable region described by sequence; ii) The heavy chain variable region described in the amino acid sequence of SEQ ID NO: 67 and the amino acid sequence of SEQ ID NO: 68 Antibodies or antigen-binding fragments containing a light chain variable region described by an acid sequence; iii) The heavy chain variable region described in the amino acid sequence of SEQ ID NO: 71 and the amino acid sequence of SEQ ID NO: 72 Antibodies or antigen-binding fragments containing a light chain variable region described by an acid sequence; iv) The heavy chain variable region described in the amino acid sequence of SEQ ID NO: 75 and the amino acid sequence of SEQ ID NO: 76 Antibodies or antigen-binding fragments containing light chain variable regions described by acid sequences; and v) The heavy chain variable region described in the amino acid sequence of SEQ ID NO: 79 and the amino acids of SEQ ID NO: 80 An antibody or antigen-binding fragment containing a light chain variable region described by sequence.

[0041] In this invention, the term "percent" is used in relation to amino acid sequences and / or nucleic acid sequences. "identity", "sequence identity", "percentage similarity", "sequence similarity", "percentage sequence identity" These methods maximize the similarity between aligned amino acid residues or nucleotides, ensuring they are the same or similar. The number of residues or nucleotides, the total number of residues or nucleotides, and the gaps in sequence alignment. The degree of similarity between two arrays is determined by comparing the sorting of arrays, which is a function of existence and length. It can refer to a defined scale, but is not limited to it.

[0042] A portion of a polynucleotide or polypeptide sequence is used as a reference for optimal alignment of two sequences. A row (without addition or fruiting) includes addition or fruiting (i.e., a gap). This is possible. The percentages mentioned above refer to the positions where the same nucleic acid base or amino acid residue appears in both sequences. Determine the number of matching positions and calculate the number of matching positions. The sequence identity percentage is calculated by dividing the result by the total number of positions on the plane and multiplying the result by 100. It is calculated.

[0043] The terms "identical" or "identical" as a percentage of two or more nucleic acid or polypeptide sequences are used in relation to two or more nucleic acid or polypeptide sequences. BLAST or BLAST 2.0 using the default parameters described below. When measuring using an array comparison algorithm, or by passive alignment and visual inspection, the same or the specified percentage of the same amino acid residue or nucleotide (i.e., comparison screen) Or, when compared and aligned for the greatest degree of correspondence to the displayed area, the concrete Approximately 60% identity for the region, preferably 65%, 70%, 75%, 80%, 85%. 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% Refers to two or more sequences or subsequences having (or higher identity) (e.g., NCB) Website: http: / / www.ncbi.nlm.nih.gov / BLAS (See T / , etc.). Furthermore, such sequences are referred to as "substantially identical."

[0044] In this invention, the amino acid sequence or base sequence is the amino acid sequence represented by each sequence number. Or, each of the base sequence is 70% or more, preferably 80% or more, more preferably 90% Most preferably, the base sequence may include a sequence having 95% or more sequence homology. For example, 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%, Sequences with 99% or 100% sequence homology may be included, but are not limited to this. No.

[0045] In this invention, naturally occurring "antibodies" are linked by disulfide bonds between chains, and It is a glycoprotein containing two heavy chains (H) and two light chains (L). Each heavy chain is, It consists of a heavy chain variable region (VH) and a heavy chain invariant region. The heavy chain invariant region consists of three domains. It consists of CH1, CH2, and CH3. Each light chain has a variable light chain region (abbreviated as VL). It consists of a light chain invariant region and a light chain invariant region. The light chain invariant region is composed of domain CL.

[0046] The VH and VL domains are separated by a more conservative domain called the Framework Domain (FR). It is further subdivided into highly variable regions called separated complementarity determination regions (CDRs). It is possible. Each VH and VL has FR1, C from the amino terminus to the carboxyl terminus. Three CDRs arranged in the order DR1, FR2, CDR2, FR3, CDR3, FR4 It is composed of four FRs. The variable regions of the heavy and light chains are binding domains that interact with the antigen. It includes n.

[0047] In this invention, antibodies can be derived from a variety of species, such as mice, rats, rabbits, and guinea pigs. However, it is not limited to this. Also, a species that has an antigen-binding site from another species It contains a chimeric antibody having an antibody-invariant region from a species (preferably). The antibody is of origin Antibodies from non-human species have antigen-binding sites that possess an invariant region and a framework region. Contains antibodies. The antibodies described herein are monoclonal antibodies, that is, they have a single molecular composition. It may be an antibody molecule, but is not limited to it. Monoclonal antibodies Systems, immunological procedures, and techniques for producing and isolating secreting hybridomas are our business. It is common knowledge in the world.

[0048] In this invention, "antigen" refers to a substance containing a binding site (epitope), and prior It induces and / or produces an immune response against the epitope. Antigens are peptides, proteins. It may include lipids, glycoproteins, polysaccharides and lipids, parts thereof and combinations thereof. However, this is not limited to these. Non-restrictive exemplary antigens include tumor antigens or pathogenic antigens. It includes the origin. Furthermore, the term "antigen" can refer to a molecule that triggers an immune response. Yes. Such immune responses include all of the following: antibody production or activation of specific immunoactive cells. In effect, any macromolecule, including virtually all proteins or peptides, can be used as an antigen. Those skilled in the art should understand that it can be used.

[0049] In this invention, the "binding site" or "epitope" is an antigen-determining cluster in a molecule, i.e., an immunoglobulin. This can refer to the part of a molecule that is recognized by an infectious agent (for example, by an antibody), but It is not limited to this. In the present invention, the epitope is DR3 and TL1A or DR3 and - It can mean including, but is not limited to, the region to which the DR3 antibody binds. There isn't one.

[0050] In one embodiment of the present invention, the antibody or the antigen-binding fragment is the amino acid of SEQ ID NO: 83. It may contain the heavy chain described in the sequence and the light chain described in the amino acid sequence of SEQ ID NO: 84. It is possible, but it is not limited to this.

[0051] The 4-4 IgF4 antibody, which has been confirmed to be the most effective in this invention, is used for graft-versus-host disease. As a result of manufacturing for application to autoimmune diseases or type 2 diabetes, one embodiment of the present invention In the example, the antibody or the antigen-binding fragment may contain Fab, but is limited to It is not limited.

[0052] Therapeutic antibodies are typically immunoglobulin G (IgG), and IgG is... It can be divided into four subclasses (IgG1, IgG2, IgG3, IgG4). The subclasses of gG have different effector functions. Therefore, it is important to select the appropriate IgG subclass based on the indication for the therapeutic antibody. This is important. In this laboratory, the effector f is determined by the IgG class. In addition to the binding action, the antibody's binding affinity to the target antigen and its activation function differ. This was confirmed to be the case. Furthermore, this is determined by the IgG subclass according to the type of antigen-antibody. The strength of the bonding affinity may appear differently depending on the material.

[0053] IS, which is functionally superior to 4-4, which was confirmed to be the most effective in this invention. As a result of confirming the OTYPE, in one embodiment of the present invention, the antibody or the antigen-binding fragment This may be an IgG4 subclass, but is not limited to it. isn't it.

[0054] In one embodiment of the present invention, the antibody or the antigen-binding fragment is IgG1 or IgG4 It may be a subclass, but is not limited to that.

[0055] In the present invention, "antigen-binding fragment" is a fragment of an antibody that possesses specific antigen-binding ability, for example. For example, (i) a monovalent fragment composed of a Fab fragment, VL, VH, CL and CH domain; ii) F(ab')2 fragment, two Fab fragments linked by disulfide bonds in the hinge region. (iii) A bivalent fragment containing the VH and CH domains; (iv) an antibody Fv fragment consisting of a single arm VL and VH domains; (v) VH domain (vi) isolated single-domain antibody fragments; (v) isolated complementation-determining regions (CDRs), and (v ii) A combination of two or more isolated CDRs selectively linked by a synthetic linker. This refers to one or more fragments of an antibody that contain a binding fragment containing a synthetic linker. - A recombination method is used to link the two domains VL and VH of the FV segment. There are single-chain antibodies (or single-chain Fv(scFv)) formed as a result.

[0056] In one embodiment of the present invention, the antigen-binding fragment is Fv, Fab, F(ab')2, Fa From b', dsFv, scFv, sc(Fv)2 and diabody You may choose any one of the following groups, but you are not limited to this.

[0057] In this invention, an agonistic anti-D for activating the target antigen human DR3 is used. Epitope mapping is used to develop R3 antibodies. It has a binding site that overlaps with TL1A, a ligand that activates human DR3 through [a specific mechanism]. Upon discovering anti-DR3 antibodies, it was found that the antibody or antigen-binding fragment of the present invention is TL1A and DR The epitopes that bind to 3 may be identical or superimposed, in which case they competitively bind specifically to DR3. This can induce regulatory T cell activity, but is not limited to this. Furthermore, if the epitopes that bind to TL1A and DR3 do not overlap, then TL1A and DR3 It can bind to DR3 and activate DR3 effectively even in a non-competitive manner. Therefore, the anti -DR3 antibodies bind to DR3 competitively and / or non-competitively with TL1A to enhance the function of DR3. Activating this process can prevent or treat graft-versus-host disease, autoimmune diseases, or type 2 diabetes. It can show the results.

[0058] In one embodiment of the present invention, the antibodies or antigen-binding fragments described in iii), iv) and v) are T While L1A can specifically bind to DR3 in a competitive manner, it is not limited to this. That is, iii), iv) and v) the binding site of the antibody or antigen binding fragment to DR3 ( The epitope may be superimposed on the binding site of TL1A to DR3, but is not limited to this. It is not something that should be done.

[0059] In one embodiment of the present invention, the antibody or the antigen-binding fragment is one of the following: The above characteristics may apply, but are not limited to: Selectively activate regulatory T cells or increase their survival rate; and Activates NF-κB.

[0060] In this invention, the anti-DR3 antibody is approximately 10 times the concentration of CD3+CD4+NF-κB. It can exhibit an activation efficiency of %~80%, and for CD3+CD8+NF-κB it is approximately 1~ While it can exhibit an activation efficiency of 60%, it is not limited to this. In particular, this development Akira's anti-DR3 4-4 IgG1 antibody is CD3+CD4+NF-κB and CD3+C Depending on the antibody treatment concentration, the phosphate levels of D8+NF-κB can be up to approximately 80% and 60%, respectively. In that it induces the chemical process, it may increase the activity efficiency of NF-κB cells through this process. However, it is not limited thereto. In this invention, the anti-DR3 antibody is CD3 + CD4 + NF- For κB, 10% for exceeding 10, 99% for exceeding 10, 98% for exceeding 10, 97% for exceeding 10. 96% over 10, 95% over 10, 94% over 10, 93% over 10, 92% over 10, 91% over 10, 90% over 10, 89% over 10, 88% over 10, 87% over 10, 86% over 10, 85% over 10, 84% over 10, 83% over 10, 82% over 10, It can exhibit an activation efficiency of 81% or more than 10, and preferably even if it is 80% or more than 10. However, this is not the limiting factor. Also, anti-DR3 antibodies are CD3 + CD8 + N For F-κB, 100% over 0, 95% over 0, 90% over 0, 85% over 0, over 0 Over 80%, Over 0 75%, Over 0 70%, Over 0 69%, Over 0 68%, Over 0 67%, 0 exceeded 66%, 0 exceeded 65%, 0 exceeded 64%, 0 exceeded 63%, 0 exceeded 62%, 0 exceeded 61 It can show an activation efficiency of % and preferably may be greater than 0 or more than 60%, but this It is not restricted.

[0061] In this invention, the anti-DR3 antibody can induce potent expansion of regulatory T cells in vivo. For example, the number and percentage of regulatory T cells can increase in the lymph nodes and spleen. This may, but is not limited to, regulatory T cells.

[0062] In this invention, the anti-DR3 antibody is more effective against DR3 than TL1A, which is a ligand for DR3. The present invention exhibits excellent bonding strength, resulting in a remarkably low bond dissociation constant. Anti-DR3 antibody showed approximately 2 times greater than 10,000 times greater than TL1A, and approximately 9,800 times greater than 2 times greater than 10,0 Approximately 9,600 times more than approximately 2 times, approximately 9,400 times more than approximately 2 times, approximately 9,200 times more than approximately 2 times, approximately 4 times Exceeded by approximately 10,000 times, exceeded approximately 4 times by approximately 9,800 times, exceeded approximately 4 times by approximately 9,600 times, exceeded by approximately 4 times About 9400 times, about 4 times more than about 9200 times, about 6 times more than about 10000 times, about 6 times more than about 98 00 times, about 6 times more than about 9600 times, about 6 times more than about 9400 times, about 6 times more than about 9200 times, About 8 times more than about 10000 times, about 8 times more than about 9800 times, about 8 times more than about 9600 times, about 8 times more Exceeded approximately 9400 times, approximately 8 times exceeded approximately 9200 times, exceeded approximately 10 times approximately 10000 times, approximately 10 times exceeded Excess of approximately 9800 times, Exceeding approximately 10 times approximately 9600 times, Exceeding approximately 10 times approximately 9400 times, Exceeding approximately 10 times Approximately 9200 times, approximately 12 times exceeded Approximately 10000 times, approximately 12 times exceeded Approximately 9800 times, approximately 12 times exceeded It can exhibit a bonding strength of approximately 9600 times, more than approximately 12 times, and more than approximately 9400 times, preferably about 1 It can exhibit a bonding strength of more than twice, approximately 9200 times or more, but is not limited to this. do not have.

[0063] The present invention provides a nucleic acid sequence for encoding the antibody or antigen-binding fragment.

[0064] In this invention, "genome" can mean the genetic material of an organism (e.g., chromosomes). "Nucleic acid sequence" or "gene" refers to polypeptides or precursors (e.g., proinsulin). Nucleic acid (e.g., DNA or RNA) sequences containing encrypted sequences necessary for production It can be tasted, but is not limited to this.

[0065] The "nucleotide sequence" or "base sequence" of the present invention is one whose manipulation is common in the art. One of the technologies can be used for a specified reason (e.g., a disease is treated or its characteristics are improved, host cells) It is also considered preferable to express the protein of the present invention and to confer the expression of ribozymes. This refers to a certain nucleotide sequence (e.g., RNA or DNA) that can exist. The cido sequence is not limited to this, but also includes the encoded sequence of a structural gene (for example). Reporter genes, screening marker genes, oncogenes, drug resistance Sex genes, growth factors, etc.), and unencrypted data that does not encode mRNA or protein products. Nodal sequences (e.g., promoter sequences, polyadenylated sequences, termination sequences, enhancer sequences) It can include columns, etc.

[0066] The present invention provides a vector containing the nucleic acid sequence.

[0067] The "vector" of the present invention is a plasmid, phage, transposon, cosmid, chromosome, This refers to the genetic elements of viruses, virions, etc., and these require proper control. When linked together with an element, replication becomes possible, and gene sequences can be moved between cells. It is possible, but it is not limited to this.

[0068] In this invention, "recombinant expression vector" refers to bacterial plasmids, phages, yeast plasmids, and plants. This refers to cell viruses, mammalian cell viruses, or other vectors. Generally, any plant Sumids and vectors can be used if they can be replicated and stabilized within the host.

[0069] The recombinant expression vector and the expression vector containing a suitable transcription / translation regulatory signal are described by those skilled in the art. It can be constructed by methods that are well known. The said method is in vitro recombinant DNA technology, DNA This includes synthesis technology and in vivo recombination technology.

[0070] Furthermore, the vectors used for recombination include both viral and non-viral vectors. These can also be used. For example, a viral vector is a wrench. Virus (lentivirus), retrovirus, adeno Viruses (adenovirus), herpes virus, or This can be done using avipox virus vectors, etc. However, it is not limited to this.

[0071] Furthermore, tags for increasing the production of recombinant proteins, recombinant proteins Genes for tagging to maintain structural stability, and for easily isolating recombinant proteins. Genetic markers for selection, such as tags and antibiotic resistance genes for selecting transformants. Children and other sub-children may be included, but this is not the only way to do so.

[0072] The present invention provides a transformant transformed with the aforementioned vector.

[0073] In this invention, "transformation" refers to the process by which injected DNA is transformed The term "transgenic" is a general term for any change in the genetic properties of an organism. A ganism is a living organism created by injecting external genes using molecular genetic methods. Preferably, cells or living organisms transformed by the recombinant expression vector of the present invention It can mean this, but is not limited to this.

[0074] In certain cases, the aforementioned living organisms include microorganisms, eukaryotic cells, insects, animals, plants, and other living things. Any living organism may be included without restriction, preferably including Intestinal bacteria, Salmonella, and Bacillus. Yeast, animal cells, mice, rats, dogs, monkeys, pigs, horses, cattle, Agrobacterium This includes, but is not limited to, plants such as Mutumefaciens.

[0075] The aforementioned transformants are transformed, transfected. fection, Agrobacterium-mediated transformation Methods: particle gun bombardment, ultrasonic treatment Sonication, electroporation, PE Manufactured by methods such as G (Polyethylene glycol)-mediated transformation. It is possible, but not limited to, G-fectin, when introduced into cells. Mirus TransIT-TKO lipid affinity reagent, lipofectin, lipofectam Cellfectin, cationic phospholipid nanoparticles, cations A transfer reagent containing a cation polymer, cationic micelle, cationic emulsion, or liposome. They are either introduced into cells or bonded together with biocompatible polymers such as polyethylene glycol. This may increase intracellular absorption, but is not limited to this.

[0076] When transforming eukaryotic cells with the vector of the present invention, yeast (Sac) is used as the host cell. Charomyce cerevisiae), insect cells, human cells (e.g., CHO cells) Chinese hamster ovary, W138, BHK, COS-7 Cell lines such as 293, HepG2, 3T3, RIN, and MDCK, as well as plant cells, are used. Preferably, Agrobacterium can be used. .

[0077] The method for delivering the vector into a host cell according to the present invention is, when the host cell is a prokaryotic cell, Ca Cl2 method, Hanahan method (Hanahan, D, J Mol Biol, 166:55 This can be carried out by methods such as 7-580 (1983) and electroporation. Also, if the host cell is true For nuclear cells, microinjection, calcium phosphate precipitation, electroporation, and liposomes are used. -Via vector-borne infection, DEAE-dextran treatment, and gene bombardment, etc. The riverator can be injected into the host cell.

[0078] The present invention is specific to the cell death receptor 3 (DR3). A transplantation-associated host disease comprising an anti-DR3 antibody that binds to or an antigen-binding fragment thereof as an active ingredient. The present invention provides a pharmaceutical composition for the prevention or treatment of autoimmune diseases or type 2 diabetes.

[0079] In one embodiment of the present invention, the antibody or the antigen-binding fragment is the amino acid of SEQ ID NO: 83. It may contain the heavy chain described in the sequence and the light chain described in the amino acid sequence of SEQ ID NO: 84. It is possible, but it is not limited to this.

[0080] In one embodiment of the present invention, the autoimmune disease is an autoimmune disease caused by graft-versus-host disease. Hemophagocytic lymphohistiocytosis cytosis, systemic lupus erythematosus hematosus, Kikuchi disease, adult-onset Still's disease Behcet's disease, IgG4 -Related diseases, type 1 diabetes, systemic sclerosis , psoriasis, multiple sclerosis, and Graves' thyroid Selected from a group consisting of Graves hyperthyroidism You may include one or more of these, but you are not limited to this.

[0081] The "pharmaceutical composition" according to the present invention is a suitable carrier and excipient typically used in the manufacture of a pharmaceutical composition. The excipients may further include modifiers and diluents. The excipients are, for example, diluents, binders, and disintegrants. Composed of destructive agents, lubricants, adsorbents, humectants, film-coating materials, and controlled-release additives. It may be one or more selected from the group.

[0082] The pharmaceutical compositions according to the present invention can be prepared by conventional methods as powders, granules, and sustained-release granules. Enteric-coated granules, liquids, eye drops, Elsylic preparations, emulsions, suspensions, alcoholic preparations, lozenges Aromatic perfumes, limonades, tablets, sustained-release tablets, enteric-coated tablets, sublingual tablets, hard capsules, Soft capsules, sustained-release capsules, enteric-coated capsules, pills, tinctures, softened extracts , dry extract preparations, liquid extract preparations, injectable preparations, capsules, perfusion solutions, ointments, lotions, In the form of topical preparations such as pastes, sprays, inhalants, patches, sterile injections, or aerosols. The topical preparation may be used in the form of a cream, gel, patch, spray, etc. Dosage forms such as ointments, plasters, lotions, liniments, pastes, or cataplasms preparations. It may have.

[0083] Examples of carriers, excipients, and diluents that may be included in the pharmaceutical composition according to the present invention include lactose, Dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, Erythritol, maltitol, starch, acacia gum, alginate, gelatin, Calcium phosphate, calcium silicate, cellulose, methylcellulose, amorphous Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propyl hydroxy Examples include roxybenzoate, talc, magnesium stearate, and mineral oil.

[0084] When formulating, the fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants that are typically used are... It is prepared using diluents or excipients such as fermenting agents.

[0085] As an additive to tablets, powders, granules, capsules, pills, and lozenges according to the present invention, Sorghum starch, potato starch, wheat starch, lactose, sucrose, glucose, fructose, D-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, monohydrogen calcium phosphate Um, calcium sulfate, sodium chloride, sodium bicarbonate, refined lanolin, microcrystalline cereal Lullose, dextrin, sodium alginate, methylcellulose, carboxymethyl Sodium cellulose, kaolin, urea, colloidal silica gel, hydroxypropyl sulfate Tarch, Hydroxypropyl Methylcellulose (HPMC) 1928, HPMC 2208 HPMC2906, HPMC2910, propylene glycol, casein, calcium lactate Excipients such as um, primozel; gelatin, acacia gum, ethanol, agar powder, acetic acid Cellulose talate, carboxymethylcellulose, carboxymethylcellulose calcium Ingredients: glucose, purified water, sodium caseinate, glycerin, stearic acid, carboxymethylcellulose Sodium methylcellulose, sodium methylcellulose, methylcellulose, microcrystalline cellulose Lullose, dextrin, hydroxycellulose, hydroxypropyl starch, hydroxycellulose Xymethylcellulose, purified shellac, starch paste, hydroxypropylcellulose, Hydroxypropyl methylcellulose, polyvinyl alcohol, polyvinylpyrrolidone Any binder can be used, such as hydroxypropyl methylcellulose or corn starch. Agar powder, methylcellulose, bentonite, hydroxypropyl starch, carboxy Sodium methylcellulose, sodium alginate, carboxymethylcellulose Cium, calcium citrate, sodium lauryl sulfate, anhydrous silicic acid, 1-hydroxypropyl sulfate Polycellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde Processed casein and gelatin, alginic acid, amylose, guar gum Baking soda, polyvinylpyrrolidone, calcium phosphate, gelled starch, gum arabic, amylose Pectin, pectin, sodium polyphosphate, ethylcellulose, sucrose, magnesium silicate Disintegrants such as aluminum ammonium, D-sorbitol solution, and hard anhydrous silicic acid; stearate Lucium, magnesium stearate, stearic acid, hydrogenated vegetable oil (Hydrogen (Aged vegetable oil), talc, lycopodium, kaolin, petrolatum, steamic acid Sodium phosphate, cocoa butter, sodium salicylate, magnesium salicylate, poly Teylene glycol 4000, PEG6000, liquid paraffin, hydrogenated soybean oil (Lub ri wax), aluminum stearate, zinc stearate, sodium lauryl sulfate Magnesium oxide, macrogol, synthetic aluminum silicate, Anhydrous silicic acid, higher fatty acids, higher alcohols, silicone oil, paraffin oil, polyethylene Glycol fatty acid ether, starch, sodium chloride, sodium acetate, sodium oleate Lubricants such as thorium, dl-leucine, and hard anhydrous silicic acid may be used.

[0086] The additives for the liquid formulation according to the present invention include water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, and monosulfate. Sucrose thearates, polyoxyethylene sorbitol fatty acid esters (Twin E Sterls, polyoxyethylene monoalkyl ethers, lanolin ethers, lanolin Esters, acetic acid, hydrochloric acid, aqueous ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide Thorium, prolamine, polyvinylpyrrolidone, ethylcellulose, carboxymethyl Sodium cellulose and similar materials may be used.

[0087] The syrup preparation according to the present invention may use a solution of sucrose, other sugars, or sweeteners, and as needed Depending on the application, fragrances, colorants, preservatives, stabilizers, suspending agents, emulsifiers, and viscosity modifiers may be used. obtain.

[0088] The emulsion of the present invention may use purified water, and may optionally contain emulsifiers, preservatives, stabilizers, and fragrances. Agents or other substances may be used.

[0089] The suspension according to the present invention contains acacia, tragacantha, methylcellulose, and carboxymethylcellulose. Alginic cellulose, sodium carboxymethylcellulose, microcrystalline cellulose, alginic acid Sodium, hydroxypropyl methylcellulose, HPMC1828, HPMC290 6. Suspending agents such as HPMC2910 may be used, and surfactants, preservatives, etc. may be used as needed. Stabilizers, colorants, and fragrances may be used.

[0090] The injectable preparations according to the present invention include distilled water for injection, 0.9% sodium chloride injection solution, and Ringer's injection solution. Solution, dextrose injection solution, dextrose + sodium chloride injection solution, P-Easy (P EG), Lactating Ringer's Injection, Ethanol, Propylene Glycol, Non-volatile Oil - Sesame Oils, cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, iso myristate Solvents such as propyl benzoate and benzene benzoate; sodium benzoate, sodium salicylate Sodium acetate, urea, urethane, monoethylacetamide, porcine, propyl Glycol, twin compounds, dijontinamide, hexamine, dimethylacetamide Solubilizers such as; weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ant Monia and ammonium acetate), organic compounds, proteins, albumin, peptone, gum Buffering agents such as sodium chloride; isotonic agents such as sodium sulfite (NaHSO3) ), carbon dioxide gas, sodium metabisulfite (Na2S2O5), sodium sulfite ( Stabilizers such as Na2SO3, nitrogen gas (N2), and ethylenediaminetetraacetic acid; sodi Umbi sulfide 0.1%, sodium formaldehyde sulfoxylate, thiourea , such as disodium ethylenediaminetetraacetate, sodium acetone bisulfite Sulfating agents: benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, gluconyl alcohol Pain relievers such as calcium phosphate; CMC sodium, sodium alginate, Twin8 It may contain a suspending agent such as aluminum monostearate.

[0091] The suppositories according to the present invention contain cocoa butter, lanolin, vitepsol, and polyethylene glycol. , glycerol gelatin, methylcellulose, carboxymethylcellulose, stearic acid and Oleic acid mixture, subanal, cottonseed oil, peanut oil, coconut oil, cocoa Butter + cholesterol, lecithin, lanette wax, glycerol monostearate Twin or span, Imhausen, monolene (monostearic acid) Propylene glycol, glycerin, Adeps solidus ), Buytyrum Tego-G, Cebe Spama 16 s Pharma 16), Hexalide Base 95, Cotomar, Hyd Rocote SP, S-70-XXA, S-70-XX75 (S-70-XX95), Hydro Hydrokote 25, Hydrocoat 711, Idropostol postal), Massa estrarium (A, AS, B) , C, D, E, I, T), Masa-MF, Maspol, Maspol-15, Neospostal -En, Paramound-B, Sposiro (OSI, OSIX, A, B, C, D, H, L), Suppository base type IV (AB, B, A, BC, BBG, E, BGF, C, D, 299), Postal (N, Es), Wecobi (W, R, S, M, Fs), Tezestr triglyceride Bases such as TG-95, MA, and 57 may be used.

[0092] Solid dosage forms for oral administration include tablets, pills, powders, granules, and capsules. Such solid preparations contain the extract with at least one excipient, for example, starch. Calcium carbonate, sucrose It is prepared by mixing rose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stylate talc are also used.

[0093] Liquid formulations for oral administration include suspensions, liquid preparations, emulsions, and syrups. However, in addition to water and liquid paraffin, which are commonly used simple diluents, various excipients are used, for example For example, humectants, sweeteners, fragrances, and preservatives may be included. Preparations for parenteral administration include: This includes sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. As a solvent or suspension solvent, propylene glycol is used. Polyethylene glycol, vegetable oils such as olive oil, ethyl oleate, and other injectable substances Esters and similar substances can be used.

[0094] The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. "An effective dose" means treating the disease with a reasonable benefit / risk ratio applicable to medical treatment. This means a sufficient amount, and the effective dose level depends on the patient's disease type, severity, drug activity, and the drug itself. This includes sensitivity to the drug, administration time, route of administration and elimination ratio, duration of treatment, and drugs used concurrently. It can be determined by factors and other factors well known in the medical field.

[0095] The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent, or in combination with other therapeutic agents. It may be administered sequentially or simultaneously with conventional therapeutic agents, either as a single or multiple dose. It is permissible to consider all the aforementioned factors and obtain the maximum effect with the minimum amount without side effects. It is important to administer a certain amount, which is something that is common knowledge for an expert in the art to which this invention belongs. This can be easily determined by [the method described].

[0096] The pharmaceutical compositions of the present invention can be administered to individuals via various routes. All methods of administration are predetermined. It is conceivable, for example, oral administration, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, perispiratory administration Intradural injection, sublingual administration, buccal administration, rectal insertion, vaginal insertion, ocular administration, ear administration, It can be administered by nasal administration, inhalation, spraying through the mouth or nose, skin administration, transdermal administration, etc. .

[0097] The pharmaceutical composition of the present invention is determined by the disease to be treated, the route of administration, the patient's age, sex, weight, and the disease. It is determined by the type of drug, which is the active ingredient, along with various related factors such as severity. Specifically, the effective amount of the composition according to the present invention varies depending on the patient's age, sex, and weight. This can be done, and generally, 0.001 to 150 mg per kg of body weight, preferably 0.01 It can be administered at a dose of up to 100 mg daily or every other day, or in 1 to 3 divided doses per day. However, the dosage may be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age, etc. The dosage, by any means, does not limit the scope of this invention.

[0098] In this invention, "individual" means a person who requires treatment for a disease, and more specifically, a human or Non-human primates, mice, rats, dogs, cats, horses and other animals It refers to mammals such as crickets.

[0099] In this invention, "administration" means providing a predetermined composition of the present invention to an individual by any appropriate method. This means that in this invention, "prevention" means suppressing and delaying the onset of the target disease. It means all actions, and "treatment" means the administration of the pharmaceutical composition according to the present invention to achieve the intended purpose. This refers to all actions that improve or favorably alter the symptoms of a disease and its associated metabolic abnormalities. "Improvement" refers to the parameters related to the target disease as a result of administering the composition according to the present invention. —For example, it means any action that reduces the severity of symptoms.

[0100] The present invention relates to graft-versus-host disease, autoimmunity, and the vector or composition, and manual, including the vector or composition, and manual. To provide a kit for the prevention or treatment of infectious diseases or type 2 diabetes.

[0101] In addition to the above configuration, the "kit" of the present invention also includes a method for expressing the vector of the present invention, Other components, devices, substances, etc., that are normally necessary for the storage, management, and enhancement of the composition of the invention. It can include. Also, all components included in the kit can be used more than once, without limit on the number of uses. They can be used, there are no restrictions on the order in which each substance is used, and the application of each substance proceeds simultaneously. You may do so, or you may proceed to the end.

[0102] The kit of the present invention may include a container in addition to the formulation and manual. The container may serve to package the aforementioned components, and may also serve to store and secure them. i. The material of the container may be, for example, a bottle, a tube, a sachet, a seal It can take the form of an envelope, tube, ampoule, etc. These can be partially or entirely made of plastic, glass, paper, foil, or wax. It may be formed from materials such as S. The container is initially part of the container, or mechanically, adhesively Alternatively, a stopper that can be attached to the container by other means, and that is completely or partially separable, may be fitted. It is possible to do so, and a stopper can be attached that allows access to the contents by the injection needle. The set may include external packages, and the external packages relate to the use of the components. Instructions may be included, but are not limited to, those included. <Modes for carrying out the invention>

[0103] The following are preferred embodiments to aid in understanding the present invention. However, the following embodiments are The following examples are provided solely to facilitate understanding of the present invention. The content of the explanation is not limited. [Examples]

[0104] <Example 1. Discovery of anti-DR3 clones using a phage display> We used Phage display to discover a clone that specifically binds to DR3. First, the human scFv library and recombinant DR3 (recombinant scFv cl) is connected to DR3 through three panning operations using t DR3. One was selectively amplified. At each stage, the phage obtained after panning was used to confirm that the scFv clones that bind to recombinant DR3 were amplified through ELISA Subsequently, phage was infected into the single clones confirmed through colony PCR, and single clones that bind to DR3 were confirmed through ELISA. The CDR regions of each single clone were confirmed through sequencing.

[0105] As a result, five clones that bind to DR3 were discovered (Figure 1(a) and Figure 1(b) ), and the five antibodies were named 2-10, 3-18, 4-4, 6-2, and 6-15, respectively. The five antibodies were sequenced to sequence the complementarity-determining regions (CDRs) of each clone. Tables 1 and 2 show the amino acids and base sequences of CDR1 to CDR3 in the heavy chain region and variable region of each antibody. Table 3 shows the base sequences and amino acid sequences of the heavy chain region and variable region of each antibody. In each table, bold type indicates the CDR sequences (sequences) according to IMGT, and the underlined part indicates the CDR sequences (sequences) according to Kabat.

[0106]

Table 1

[0107]

Table 2

[0108]

Table 3-1

[0109] ​[Table 3-2]

[0110] [Table 3-3]

[0111] <Example 2. Confirmation of the excellent DR3-specific binding affinity of five antibodies> The specific binding affinity of the five antibodies developed in Example 1 to DR3 was measured by ELISA, SPR, and This was confirmed through FACS experiments.

[0112] Example 2-1. Confirmation of specific binding affinity to DR3 using ELISA. ELISA was performed using the monoclonal antibody of the clone selected in Example 1. Specifically, after adding 0.4 μg / ml of DR3-His tag to PBS and mixing, A total of 50 μl of the mixture was coated onto a Ni plate. Subsequently, nonspecific binding (n To prevent on-specific binding, 200 μl of 5% FB The PBS containing S was treated and incubated at room temperature for 30 minutes to determine the concentration of the monoclonal antibody. The DR3 was titrated and incubated for 30 minutes. The degree of binding of the monoclonal antibody was detected using anti-human IgG Fc HRP. Each plate was washed five times with 250 μl of 0.1% Tween 20. An isotype control was used as the negative control group.

[0113] As a result, the five antibodies of the present invention exhibited superior specific binding affinity to DR3 compared to the control group, Among them, 4-4, 6-2, and 6-15 exhibit binding specificity to DR3 even at low antibody concentrations. It was confirmed that the levels remained remarkably high (Figure 2(a)).

[0114] Example 2-2. Confirmation of specific binding affinity to DR3 using SPR. SPR was carried out using the monoclonal antibodies of the clones selected in Example 1. C After immobilizing 10 μg / ml of DR3-His tag on the M5 chip and allowing a contact time of 30 sec and a flow rate of 30 μl / min, the binding specificity and its binding affinity were measured using a Biacore T100 .

[0115] As a result, the five antibodies according to Example 1 had a significantly lower dissociation constant for binding to DR3 than that of the DR3 agonist TL1A, and it was confirmed that they showed a binding affinity of about 12 to over 9,20 times (Fig. 2(b)).

[0116] Example 2-3. Confirmation of specific binding affinity to DR3 using FACS. FACS was carried out using the monoclonal antibodies of the clones selected in Example 1. Specifically, 0.2 X 10 6 of the DR3 stable cell line was mixed with 2 μg / ml of the monoclonal antibody and incubated at 4°C for 30 minutes. The degree of binding between the DR3 stab le cell line and the monoclonal antibody was confirmed through FACS analysis using anti-human IgG PE . The cells were washed once with 200 μl of PBS at each step . An isotype control was used for the negative control group

[0117] As a result, it was confirmed that antibodies 4-4, 6-2, and 6-15 strongly bind to DR3 (Fig 2(c)).

[0118] <Example 3. Epitope mapping of five antibodies against DR3> Example 3-1. Epitope mapping using ELISA ​ (ing) Of the five antibodies in Example 1, the rDR3 epitope to which the anti-DR3 4-4 antibody binds is r To confirm whether DR3 is superimposed on TL1A, its ligand, or other monoclonal molecules. An ELISA was performed. Specifically, a 0.4 μg / ml DR3-His tag was used in PBS. After mixing in the container, a total of 50 μl of the mixture was coated onto a Ni plate. Furthermore, to prevent non-specific binding, 20 Mix with 0 μl of PBS containing 5% FBS and incubate at room temperature for 30 minutes. The concentrations of TL1A and monoclonal antibody were titrated and incubated for 30 minutes. I then applied 200 ng / ml of biotinylated antibiotic without washing. -Incubate DR3 4-4 antibody for 30 minutes, and combine DR3 and biotinylate The degree of binding of anti-DR3 4-4 antibody was detected by anti-human IgG Fc HRP. Except for the stage without washing, the plate was treated with 250 μl of 0.1% Twe at each stage. Washing was performed 5 times with EN 20. The negative control group was an isotype control. I used it.

[0119] Example 3-2. Epitope mapping using FACS ng) The epitope to which five selected antibodies bind to DR3 expressed on the cell surface is human DR3. To confirm whether it superimposes with TL1A, which is its ligand, or other monoclonal antibodies. FACS was performed. 0.2X10 6 2μ of DR3 stable cell line Mix with g / ml monoclonal antibody and incubate at 4°C for 30 minutes, then wash. Mix 0.625 μg / ml of TL1A His tag without adding any other ingredients and incubate at 4°C for 30 minutes. Incubation was performed. DR3 stable cell line and TL1A-His The degree of tag binding is detected using anti-His tag AF647 and then passed through FACS. I confirmed this. Except for the stage without washing, the cells were in 200 μl PB at each stage. A single wash was performed with S. The negative control group used an isotype control. .

[0120] As a result, clones 4-4, 6-2, and 6-15 form epitones that bind DR3 and TL1A. While they bind to the epitope, clones 2-10 and 3-18 do not bind to the epitope. This was confirmed (Figures 3(a) and 3(b)).

[0121] <Example 4. Confirmation of excellent NF-κB activity effect by selected antibodies> To confirm the NF-κB activity effect of the antibodies selected in Example 1, phosphorylation was performed using FACS. The NF-κB (phosphorylated NF-κB, pNF-κB) level We compared and analyzed clone4-4 IgG1 and clone2-10 IgG. 1. Clone3-18 IgG1, Clone6-2 IgG1 monoclonal antibody and Activated by rhTL1A. 1X10 6 The hPBMC also has an rhTL1A-His tag. After mixing the concentration-adjusted monoclonal antibody, incubate at 37°C for 10 minutes. Incubated. NF-κB activation level (pNF-κB) was measured against anti-human phosphate. Using FACS with pho-NF-κB p65(Ser529)eFlour 660 I analyzed it.

[0122] As a result, it was confirmed that the highest level of NF-κB activation was achieved by 4-4(IgG1). Next was 6-2. This is because the site where the antibody binds to DR3 overlaps with TL1A. It was investigated that this is because NF-κB is strongly phosphorylated while being folded (Figure 4).

[0123] <Example 5. Comparison of binding strength by IgG subclass of Clone4-4> Functionally, the anti-DR3 4-4 antibody, which was confirmed to be the most superior in Example 2, is the most effective. We also analyzed the superior subclass. For this purpose, we used an anti-DR3 4-4 antibody against IgG. Depending on the subclass, clone4-4 IgG1, clone4-4 IgG2, Prepare a clone4-4 IgG4 monoclonal antibody and determine its binding affinity to DR3. An ELISA was performed for comparison. Specifically, 0.4 μg / ml DR3-His After mixing the tag in PBS, coat the Ni plate with a total of 50 μl of the mixture. It was then performed to prevent non-specific binding. To do this, mix with PBS consisting of 200 μl of 5% FBS and incubate at room temperature for 30 minutes. Incubate, titrate with monoclonal antibody and incubate for 30 minutes. The degree of binding between the coated DR3 and the monoclonal antibody was determined to be anti-human Ig Detected by G Fc HRP. For each stage, the plate was 250 μl 0.1% Twe I washed it five times with EN 20.

[0124] As a result, the anti-DR3 4-4 antibody was found to have subclasses IgG1, IgG4, and Ig It was confirmed that G2 showed superior affinity to DR3 (Figure 5).

[0125] Example 6. Excellent NF-κB activity by Clone4-4 IgG subclass Checking the effect > We will compare NF-κB activation levels based on IgG subclasses of Clone4-4. Therefore, clone4-4 IgG1, clone4-4 IgG2, clone4-4 Perform FACS after stimulation with IgG4 monoclonal antibody. Specifically , 1X10 6 The hPBMC has an rhTL1A-His tag or concentration. After mixing the NF monoclonal antibodies, the mixture was incubated at 37°C for 10 minutes. -κB activation level is anti-human phospho-NF-κB p65(Ser529) The analysis was performed using FACS with eFlour 660.

[0126] As a result, antibody 4-4 was classified as NF in the order of subclass IgG4, IgG1, and IgG2. A high level of -κB activation was confirmed (Figure 6).

[0127] <Example 7. Regulatory T-cells of anti-DR3 4-4 IgG4 antibody in the absence of TCR signal stimulation> Effect of increasing cell survival rate > Against the anti-DR3 4-4 IgG4 antibody that was confirmed to be the best in Example 6 The antibody in question can increase the survival rate of human regulatory T cells when there is no TCR signaling stimulation. To confirm this, we used anti-DR3 4-4 IgG4 antibody to culture CD4 T cells. Nutrition was performed. Specifically, Human CD4 T cell Enrichment K After isolating CD4 T cells from hPBMCs using it, 20 ng / ml of IL-2 was added. Add 5 ng / ml of TL1A or 1 μg / ml of anti-DR3 4-4 IgG4 antibody. The samples were cultured for 3 days and then analyzed via FACS.

[0128] As a result, in an environment without TCR signal stimulation, the anti-DR3 4-4 IgG4 antibody can increase the survival rate of regulatory T cells at a level similar to that of the DR3 ligand TL1A was confirmed (Figure 7).

[0129] <Example 8. Effect of anti-DR3 4-4 IgG4 antibody on the proliferation and survival rate of regulatory T cells by TCR signal intensity> For the anti-DR3 4-4 IgG4 antibody, which was confirmed to be the most excellent in Example 6, to confirm the degree of promotion of the division of human regulatory T cells by the corresponding antibody according to the intensity of TCR signal stimulation, anti-hCD3 antibody and anti-DR3 4-4 IgG4 antibody were used for CD4 T cell culture Specifically, one day before cell culture, anti-hCD3ε antibodies at 1.5 μg / ml, 0.5 μg / ml, 0.25 μg / ml, 0.125 μg / ml, and 0 μg / ml were respectively coated on a 96-well l cell culture plate, and then on the day of cell culture, CD4 T cells were separated from hPBM C using the Human CD4 T Cell Enrichment Kit. Then, 0.1×10 CD4 6 T cells per well were cultured, and 20 ng / ml of IL-2, 0.5 ng / ml of TGF-β or anti-DR3 4-4 IgG4 antibody were added according to the group and cultured for ⑥ days, followed by analysis through FACS [[ID=3(]]

[0130] As a result, when TCR signal stimulation above a specific intensity was given, the anti-DR3 4-4 Ig G4 antibody was confirmed to effectively increase the division or survival rate of regulatory T cells (Figure 8 ).

[0131] ​​​​​ <Example 9. Sequencing of anti-DR3 4-4 IgG4 monoclonal antibody> Examples 3-8 are intended for use in the treatment of graft-versus-host disease, autoimmune diseases, and type 2 diabetes. The anti-DR3 4-4 IgG4 monoclonal has been confirmed to have the best performance. The antibody was sequenced. Specifically, based on sequence analysis data, a 4-4 mAb Fab region was selected. The gG4 expression vector was cloned, and the DNA and amino acids of the cloned vector were extracted. The acid sequences are shown in Table 4. Table 4 shows the fully human 4-4 DN. This shows the A sequence and amino acid sequence.

[0132] [Table 4]

[0133] The above description of the present invention is illustrative and does not apply to the usual practices in the art to which the present invention belongs. Anyone with knowledge of the present invention can modify other aspects without changing the technical concept or essential features of the present invention. It can be easily modified into a specific form. Therefore, the above-described embodiment is illustrative in all respects. It must be understood that this is a general concept, not a limited one. [Industrial applicability]

[0134] Antibodies that specifically bind to human DR3 are those that bind to the TL1A ligand of human DR3 via the epithelium. It specifically binds to the cytoplasm and selectively acts on regulatory T cells, thereby affecting effector CD4+ T cells. This minimizes the activity of CD8+ T cells while improving the division or survival rate of regulatory T cells. Therefore, it can be usefully utilized as a therapeutic agent for graft-versus-host disease, autoimmune diseases, or type 2 diabetes. In conclusion, it is recognized as having industrial applicability.

Claims

1. Specific binding to the cell death receptor 3 (DR3). An anti-DR3 antibody or its antigen-binding fragment characterized by the above.

2. The antibody or antigen-binding fragment is one selected from the group consisting of the following: The anti-DR3 antibody or antigen-binding fragment thereof according to claim 1, characterized by: i) The heavy chain CDR1 described in the amino acid sequence of SEQ ID NO: 1, and the amino acid sequence of SEQ ID NO: 2 Includes heavy chain CDR2 as described and heavy chain CDR3 as described in the amino acid sequence of SEQ ID NO:

3. Heavy chain variable region; and light chain CDR1 as described in the amino acid sequence of SEQ ID NO: 4, SEQ ID NO: 5 Light chain CDR2 and light chain SEQ ID NO: 6 described by amino acid sequence Antibodies or antigen-binding fragments containing a light chain variable region including CDR3; ii) The amino acid sequence of heavy chain CDR1 described in the amino acid sequence of SEQ ID NO: 7, and the amino acid sequence of SEQ ID NO: 8 It contains heavy chain CDR2 as described in and heavy chain CDR3 as described in the amino acid sequence of SEQ ID NO:

9. The heavy chain variable region; and the light chain CDR1 described in the amino acid sequence of SEQ ID NO: 10, SEQ ID NO: The light chain CDR2, described by the amino acid sequence of 11, and the amino acid sequence of SEQ ID NO: 12 An antibody or antigen-binding fragment containing a light chain variable region including a light chain CDR3; iii) The heavy chain CDR1 described in the amino acid sequence of SEQ ID NO: 13, and the amino acid sequence of SEQ ID NO: 14 Heavy chain CDR2 described by its acid sequence and heavy chain CD described by its amino acid sequence of Sequence ID No. 15 Heavy chain variable region including R3; and light chain CDR1 as described by the amino acid sequence of SEQ ID NO: 16, The amino acid sequence of the light chain CDR2 described in SEQ ID NO: 17 and the amino acid sequence of SEQ ID NO: 18 An antibody or antigen-binding fragment containing a light chain variable region including the light chain CDR3 as described above; iv) The amino acid sequence of heavy chain CDR1 described in SEQ ID NO: 19, and the amino acid sequence of SEQ ID NO: 20 Heavy chain CDR2 described in sequence and heavy chain CDR described in amino acid sequence of SEQ ID NO: 21 A heavy chain variable region containing 3; and the light chain CDR1 described in the amino acid sequence of SEQ ID NO: 22, The amino acid sequence of the light chain CDR2 described in the amino acid sequence of sequence number 23 and the amino acid sequence of sequence number 24 Antibodies or antigen-binding fragments containing a light chain variable region including the described light chain CDR3; and v) The amino acid sequence of heavy chain CDR1 described in SEQ ID NO: 25, and the amino acid sequence of SEQ ID NO: 26 Heavy chain CDR2 is described in columns and heavy chain CDR3 is described in the amino acid sequence of SEQ ID NO:

27. Heavy chain variable region including; and light chain CDR1, sequence described in the amino acid sequence of SEQ ID NO:

28. The amino acid sequence of the light chain CDR2, described as amino acid sequence number 29, and the amino acid sequence of sequence number 30 are described. An antibody or antigen-binding fragment containing a light chain variable region including the listed light chain CDR3.

3. The antibody or antigen-binding fragment is one selected from the group consisting of the following: The anti-DR3 antibody or antigen-binding fragment thereof according to claim 1, characterized by: i) The heavy chain variable region described in the amino acid sequence of SEQ ID NO: 63 and the amino acids of SEQ ID NO: 64 An antibody or antigen-binding fragment containing a light chain variable region described by sequence; ii) The heavy chain variable region described in the amino acid sequence of SEQ ID NO: 67 and the amino acid sequence of SEQ ID NO: 68 Antibodies or antigen-binding fragments containing a light chain variable region described by an acid sequence; iii) The heavy chain variable region described in the amino acid sequence of SEQ ID NO: 71 and the amino acid sequence of SEQ ID NO: 72 Antibodies or antigen-binding fragments containing a light chain variable region described by an acid sequence; iv) The heavy chain variable region described in the amino acid sequence of SEQ ID NO: 75 and the amino acid sequence of SEQ ID NO: 76 Antibodies or antigen-binding fragments containing light chain variable regions described by acid sequences; and v) The heavy chain variable region described in the amino acid sequence of SEQ ID NO: 79 and the amino acids of SEQ ID NO: 80 An antibody or antigen-binding fragment containing a light chain variable region described by sequence.

4. The antibody or antigen-binding fragment is a heavy chain and a compound described in the amino acid sequence of SEQ ID NO:

83. The claim is characterized by containing a light chain described by the amino acid sequence of column number 84. The anti-DR3 antibody described in 1, or its antigen-binding fragment.

5. The antibody or antigen-binding fragment is characterized by containing Fab, as described in claim 4. - DR3 antibody or its antigen-binding fragment.

6. The antibody or antigen-binding fragment is an IgG1 or IgG4 subclass The anti-DR3 antibody or antigen-binding fragment thereof according to claim 1, characterized in that it is the same as the anti-DR3 antibody or antigen-binding fragment thereof.

7. The antigen-binding fragments are Fv, Fab, F(ab')2, Fab', dsFv, scFv, Any of the following selected from the group consisting of sc(Fv)2 and diamond body. The anti-DR3 antibody or antigen-binding fragment thereof according to claim 1, characterized in that it is one of the two. 。

8. The antibodies or antigen-binding fragments described in iii), iv), and v) are competitive with TL1A and are particularly specific to DR3. The anti-DR3 antibody according to claim 2, characterized in that it binds heteroconjugately, or the anti-DR3 antibody according to claim 2. Original combined fragment.

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

10. A nucleic acid sequence characterized by encoding the antibody or antigen-binding fragment described in claim 1.

11. A vector characterized by comprising the nucleic acid sequence described in claim 10.

12. A transformant characterized by being transformed into the vector described in claim 11.

13. The cell death receptor 3 (DR3) described in claim 1 is specific to the cell death receptor 3. It is characterized by containing an anti-DR3 antibody or its antigen-binding fragment as an active ingredient that binds to the target antibody. A pharmaceutical composition for the prevention or treatment of graft-versus-host disease, autoimmune diseases, or type 2 diabetes.

14. The antibody or antigen-binding fragment is a heavy chain and a compound described in the amino acid sequence of SEQ ID NO:

83. The claim is characterized by containing a light chain described by the amino acid sequence of column number 84. The pharmaceutical composition described in 13.

15. The aforementioned autoimmune diseases include graft-versus-host disease, hemophagocytic lymphohistiocytosis ( Hemophagic lymphohistiocytosis, systemic erythritis Systemic lupus erythematosus, Kikuchi Adult Still's disease, Behcet's disease, IgG4-associated disease, type 1 diabetes , systemic sclerosis, psoriasis, multiple sclerosis (mu) (Little sclerosis) and Graves' hyperthyroidism (Graves One or more selected from the group consisting of (hyperthyroidism) The pharmaceutical composition according to claim 13, characterized by the following:

16. The vector according to claim 11 or the composition according to claim 13, and the manual. A kit for the prevention or treatment of graft-versus-host disease, autoimmune disease, or type 2 diabetes, characterized by the following: 。

17. Anti-DR3 inhibitors that bind to death receptor 3 (DR3) A composition containing the body or its antigen-binding fragments as an active ingredient is administered to an individual in need. A method characterized by including a step for the prevention of graft-versus-host disease, autoimmune diseases, or type 2 diabetes. This is a treatment method.

18. Anti-DR3 inhibitors that bind to death receptor 3 (DR3) A composition containing the body or its antigen-binding fragments as an active ingredient for graft-versus-host disease, autoimmune disease, or It is used for the prevention or treatment of type 2 diabetes.

19. Anti-DR3 inhibitors that bind to death receptor 3 (DR3) A composition containing the body or its antigen-binding fragments as an active ingredient for graft-versus-host disease, autoimmune disease, or This is for use in manufacturing preventive or therapeutic agents for type 2 diabetes.

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