Fusion protein comprising dual functional domain and uses thereof

Abstract

The present invention relates to a dual function domain structure and uses thereof. The dual function domain according to the present invention comprises a first domain that binds to a disease site-specific expressing protein and a second domain that binds to a disease-specific expressing protein. The first domain serves to shield the function of a drug in non-diseased regions, thereby enabling disease site-specific drug delivery. In addition, the first domain is separated by protease specifically expressed at a disease site to induce and exhibit drug activity in a disease site-specific manner. Furthermore, a multi-specific fusion protein comprising the dual functional domain structure can be designed in various forms to include various functions. Therefore, the dual function domain structure and the multi-specific fusion protein comprising same according to the present invention can be used for various applications as alternatives to conventional antibodies.

Description

Fusion protein comprising dual functional domains and uses thereof

[0001] The present invention relates to a fusion protein comprising a dual functional domain and uses thereof. Specifically, the fusion protein comprises a first domain comprising a site that specifically binds to a first target; and a second domain comprising a site that specifically binds to an immune modulator or a second target; wherein the first target is a protein overexpressed in vascular cells or basal cells at a disease site, and the second target comprises a protein overexpressed or secreted on the surface of cancer cells or immune cells.

[0002] Antibody-based therapeutics are widely used in various fields, including cancer, inflammatory diseases, infectious diseases, and immune disorders. However, clinical trials have shown limitations, including reduced efficacy due to low drug delivery rates and adverse effects caused by drug activity in normal tissues rather than the diseased site. To overcome these issues, extensive research is currently underway to develop drugs that are specifically delivered and activated at the site of disease.

[0003] Among these, research is actively being conducted to develop a system to overcome side effects caused by drug activity in non-diseased areas through disease-specific delivery and activity of drugs by binding to proteins overexpressed in vascular cells around the diseased area.

[0004] However, despite these efforts, a visible disease-specific drug delivery system and activation technology have not yet been developed, and further research is needed.

[0005] Accordingly, the inventors of the present invention have conducted repeated studies to increase the drug delivery efficiency specifically to the diseased site without affecting normal tissues, and as a result, they have confirmed that a fusion protein of a dual domain structure including a first domain that binds to a protein specifically overexpressed in vascular cells or basal cells of the diseased site and a second domain that binds to a protein specifically expressed in disease target cells accumulates specifically at the diseased site and exhibits drug activity specifically at the diseased site, thereby completing the present invention.

[0006] To achieve the above object, one aspect of the present invention provides a fusion protein comprising a first domain including a site that specifically binds to a first target; and a second domain including a site that specifically binds to an immune modulator or a second target; and a fusion protein dimer comprising the same. In this case, the first target may be a protein overexpressed in vascular cells or basal cells at a disease site, and the second target may be a protein overexpressed or secreted on the surface of cancer cells or immune cells.

[0007] Another aspect of the present invention provides a polynucleotide encoding the fusion protein or a fusion protein dimer comprising the same, a vector comprising the polynucleotide, and a cell transformed with the vector.

[0008] Another aspect of the present invention provides a method for producing a fusion protein or a fusion protein dimer, comprising the steps of culturing the transformed cell and obtaining the fusion protein or a fusion protein dimer comprising the fusion protein from the culture medium.

[0009] Another aspect of the present invention provides the fusion protein, a fusion protein dimer comprising the fusion protein, or a drug conjugate thereof (fusion protein-drug conjugate, fusion protein dimer-drug conjugate), and a pharmaceutical composition for preventing or treating a disease comprising the fusion protein as an active ingredient.

[0010] Another aspect of the present invention provides a cleavable peptide comprising any one amino acid sequence selected from the group consisting of SEQ ID NOs: 159, 160, 173 to 176 and combinations thereof.

[0011] Another aspect of the present invention provides a use of the fusion protein, a fusion protein dimer comprising the fusion protein, or a drug conjugate thereof (fusion protein-drug conjugate, fusion protein dimer-drug conjugate) for the prevention or treatment of a disease.

[0012] Another aspect of the present invention provides a method for preventing or treating a disease, comprising administering to a subject the fusion protein, a fusion protein dimer comprising the fusion protein, or a drug conjugate thereof (fusion protein-drug conjugate, fusion protein dimer-drug conjugate).

[0013] ReleaDome (Fig. 1), a dual-functional domain construct according to the present invention, has a structure in which a first domain and a second domain are connected via a linker. At this time, the first domain binds to a protein specifically expressed or exposed in vascular cells or basal cells at the disease site, and the second domain can bind to a protein overexpressed in target cells at the disease site. In addition, the linker includes a substrate sequence that can be cleaved by a specific protease specifically expressed in the microenvironment of the disease site. Therefore, the construct is a novel construct that can be specifically activated at the disease site and thus can efficiently exert a therapeutic effect. In addition, a multi-specific fusion protein comprising the dual-functional domain construct can be designed to include various functions in various forms. Therefore, the dual-functional domain construct according to the present invention and the multi-specific fusion protein comprising the same can be utilized for various purposes as a substitute for conventional antibodies.

[0014] Figure 1 is a schematic diagram of the structure of a dual-function domain (ReleaDome).

[0015] Figure 2 is a schematic diagram of a multispecific fusion protein comprising ReleaDome. Figure 2a is a schematic diagram of a basic multispecific fusion protein composed of minimal components, and Figures 2b and 2c are schematic diagrams of a multispecific fusion protein with an extended structure.

[0016] Figure 3 is a schematic diagram showing an example of a multispecific fusion protein containing ReleaDome. Figure 3a is a schematic diagram of a DVD-Ig type multispecific fusion protein containing ReleaDome and its variants, and Figure 3b is a schematic diagram of a ReleaBody containing ReleaDome. In the form of Schematic diagram of a multispecific fusion protein and its variants, and Fig. 3c is a schematic diagram of several forms of multispecific fusion proteins including ReleaDome.

[0017] Figure 4 shows the analysis results for a DVD-Ig type multispecific fusion protein and its variants including ReleaDome [anti-ANXA1-anti-HER2], which is an example of the present invention. Figures 4a to 4e show the productivity, characteristics, and purity analysis results through SEC-UPLC (Size-exclusion-UPLC) of the DVD-Ig type multispecific fusion protein and its variant molecules. Figures 4f and 4g show the results of Coomassie Brilliant Blue staining after SDS-PAGE under reducing conditions for the DVD-Ig type multispecific fusion protein and its variants.

[0018] Figure 5 shows the analysis results of a DVD-Ig type multispecific fusion protein and its variants including ReleaDome [anti-ANXA1-anti-CD47], which is an example of the present invention. Figures 5a and 5b show the productivity, characteristics, and purity analysis results through SEC-UPLC of the DVD-Ig type multispecific fusion protein and its variant molecules. Figure 5c shows the results of SDS-PAGE under reducing conditions for the DVD-Ig type multispecific fusion protein and its variants, followed by Coomassie Brilliant Blue staining.

[0019] Figure 6 shows the results of analysis of a DVD-Ig type multispecific fusion protein and its variants including ReleaDome [anti-ANXA1-anti-EGFR], which is an example of the present invention. Figures 6a to 6c show the results of productivity, characteristics, and purity analysis through SEC-UPLC of a DVD-Ig type multispecific fusion protein and its variant molecules. Figures 6d and 6e show the results of SDS-PAGE under reducing conditions for a DVD-Ig type multispecific fusion protein and its variant molecules, followed by Coomassie Brilliant Blue staining.

[0020] Figure 7 shows the results of analysis of a DVD-Ig type multispecific fusion protein and its variants including ReleaDome [anti-ANXA1-anti-DLK1], which is an example of the present invention. Figures 7a and 7b show the results of productivity, characteristics, and purity analysis through SEC-UPLC of the DVD-Ig type multispecific fusion protein and its variant molecules. Figure 7c shows the results of SDS-PAGE under reducing conditions for the DVD-Ig type multispecific fusion protein and its variant molecules, followed by Coomassie Brilliant Blue staining.

[0021] Figure 8 shows the results of analysis of a multi-specific fusion protein in the form of ReleaBody and a variant thereof, which is an example of the present invention, including ReleaDome [anti-ANXA1-anti-HER2]. Figures 8a and 8b show the results of productivity, characteristics, and purity analysis using SEC-UPLC of the multi-specific fusion protein in the form of ReleaBody and the variant thereof. Figure 8c shows the results of SDS-PAGE under reducing conditions for the multi-specific fusion protein in the form of ReleaBody and the variant thereof, followed by Coomassie Brilliant Blue staining.

[0022] Figure 9 shows the results of analysis of a multi-specific fusion protein in the form of ReleaBody and a variant thereof, which is an example of the present invention, including ReleaDome [anti-ANXA1-anti-CTLA4]. Figures 9a and 9b show the results of productivity, characteristics, and purity analysis using SEC-UPLC of the multi-specific fusion protein in the form of ReleaBody and the variant thereof. Figure 9c shows the results of SDS-PAGE under reducing conditions for the multi-specific fusion protein in the form of ReleaBody and the variant thereof, followed by Coomassie Brilliant Blue staining.

[0023] Figure 10 shows the results of analysis of a multi-specific fusion protein in the form of ReleaBody and a variant thereof, which is an example of the present invention, including ReleaDome [anti-ANXA1-anti-CD47]. Figures 10a and 10b show the results of productivity, characteristics, and purity analysis using SEC-UPLC of the multi-specific fusion protein in the form of ReleaBody and the variant thereof. Figure 10c shows the results of SDS-PAGE under reducing conditions and Coomassie Brilliant Blue staining of the multi-specific fusion protein in the form of ReleaBody and the variant thereof.

[0024] Figure 11 shows the results of analysis of a multi-specific fusion protein in the form of ReleaBody and a variant thereof, which is an example of the present invention, including ReleaDome [anti-ANXA1-anti-EGFR]. Figures 11a and 11b show the results of productivity, characteristics, and purity analysis using SEC-UPLC of a multi-specific fusion protein in the form of ReleaBody and a variant molecule thereof. Figure 11c shows the results of SDS-PAGE under reducing conditions for a multi-specific fusion protein in the form of ReleaBody and a variant molecule thereof, followed by Coomassie Brilliant Blue staining.

[0025] Figure 12 shows the results of analysis of a multi-specific fusion protein in the form of ReleaBody and a variant thereof comprising ReleaDome [anti-ANXA1-anti-B7H3], which is an example of the present invention. Figures 12a and 12b show the results of productivity, characteristics, and purity analysis using SEC-UPLC of the multi-specific fusion protein in the form of ReleaBody and the variant thereof. Figure 12c shows the results of SDS-PAGE under reducing conditions for the multi-specific fusion protein in the form of ReleaBody and the variant thereof, followed by Coomassie Brilliant Blue staining.

[0026] Figure 13 shows the analysis results of a variant of a multispecific fusion protein [DVD-IgG CrossMab, KIH, Single arm-cleavable] including ReleaDome [anti-ANXA1-anti-EGFR], which is an example of the present invention. Figure 13a is a schematic diagram of a variant of a multispecific fusion protein [DVD-IgG CrossMab, KIH, Single arm-cleavable]. Figures 13b and 13c show the results of productivity, characteristics, and purity analysis through SEC-UPLC of a variant of a multispecific fusion protein [DVD-IgG CrossMab, KIH, Single arm-cleavable]. Figure 13d shows the results of SDS-PAGE under reducing conditions and Coomassie Brilliant Blue staining of a variant of a multispecific fusion protein [DVD-IgG CrossMab, KIH, Single arm-cleavable].

[0027] Figure 14 shows the analysis results of a variant [DVD-IgG asymmetric] of a multispecific fusion protein including ReleaDome [anti-ANXA1-anti-EGFR], which is an example of the present invention. Figure 14a is a schematic diagram of a variant [DVD-IgG asymmetric] of a multispecific fusion protein. Figures 14b and 14c show the results of productivity, characteristics, and purity analysis through SEC-UPLC of a variant [DVD-IgG asymmetric] of a multispecific fusion protein. Figure 14d shows the results of Coomassie Brilliant Blue staining after SDS-PAGE under reducing conditions of a variant [DVD-IgG asymmetric] of a multispecific fusion protein.

[0028] Figure 15 shows the results of analyzing the drug-antibody ratio (DAR) from a multi-specific fusion protein-drug conjugate containing ReleaDome. Figures 15a and 15b are examples of results analyzed using RP-UPLC (Reversed-phase chromatography), and Figures 15c to 15g are examples of results analyzed using rRP-UPLC (Reduced RP-UPLC).

[0029] Figure 16 shows the SDS-PAGE results under reducing conditions for five types of protein-decomposing enzymes.

[0030] Figures 17a to 17c show the results of analyzing the cleavable activity of a multi-specific fusion protein containing ReleaDome by MT-SP1 protease using SDS-PAGE under reducing conditions.

[0031] Figure 18 shows the results of analyzing the cleavage efficiency of a multi-specific fusion protein containing ReleaDome by uPA protease using SDS-PAGE under reducing conditions.

[0032] Figure 19 shows the results of analyzing the cleavage efficiency of a multi-specific fusion protein containing ReleaDome by human MMP-2 (hMMP-2) protease using SDS-PAGE under reducing conditions.

[0033] Figure 20 shows the results of analyzing the cleavage efficiency of a multispecific fusion protein containing ReleaDome by MMP-9 protease through SDS-PAGE under reducing conditions. Figure 20a shows an example of the results of analyzing the cleavage efficiency of a multispecific fusion protein by human MMP-9 (hMMP-9) protease through SDS-PAGE under reducing conditions. Figure 20b shows an example of the results of analyzing the concentration-dependent cleavage efficiency of hMMP-9 protease through SDS-PAGE under reducing conditions. Figure 20c shows the results of analyzing the cleavage efficiency of a multispecific fusion protein by mouse MMP-9 (mMMP-9) protease through SDS-PAGE under reducing conditions.

[0034] Figure 21 shows the results of analyzing the cleavage efficiency of the multi-specific fusion protein containing ReleaDome using the combined use of hMMP-2 and hMMP-9 proteases through SDS-PAGE under reducing conditions. Figure 21a shows the results of analyzing the cleavage efficiency of the multi-specific fusion protein, hMMP-2, and hMMP-9 (hMMP-2&-9) in a ratio of 1:0.02:0.02 and reacting through SDS-PAGE under reducing conditions. Figure 21b shows the results of analyzing the cleavage efficiency of the multi-specific fusion protein, hMMP-2, and hMMP-9 in a ratio of 1:0.02:0.06 and reacting through SDS-PAGE under reducing conditions. Figure 21c shows the results of analyzing the cleavage efficiency using SDS-PAGE under reducing conditions when multispecific fusion proteins, hMMP-2 and hMMP-9, were reacted at a ratio of 1:0.02:0.02 or 1:0.02:0.06.

[0035] Figure 22 shows the results of analyzing the antigen binding ability of multi-specific fusion proteins containing ReleaDome through ELISA. Figures 22a to 22d show the results of confirming the change in antigen [HER2] binding ability according to the concentration of each multi-specific fusion protein containing ReleaDome and the presence or absence of treatment with a protease [MT-SP1]. Figure 22e shows the results of confirming the binding ability of each multi-specific fusion protein to each antigen [ANXA1, HER2].

[0036] Figure 23 is an example of the results of mapping the antigen binding site of each UNIT1 having binding affinity to ANXA1 through ELISA.

[0037] Figure 24 shows the results of analyzing the binding ability of each tumor cell (SK-OV-3 cells, NCI-N87 cells, BT-474 cells) of a multispecific fusion protein [anti-ANXA1-anti-HER2] containing ReleaDome, which is an example of the present invention, through flow cytometry (Fc). Figure 24a shows the results of confirming the antigen binding ability according to the concentration of BA-0102-001 containing ReleaDome and the presence or absence of treatment with a protease [MT-SP1] in SK-OV-3 cells expressing HER2. Figure 24b shows the results of confirming the antigen binding ability of each multispecific fusion protein containing ReleaDome according to the concentration in NCI-N87 cells expressing HER2. Figure 24c shows the results of confirming the antigen binding ability of each multi-specific fusion protein containing ReleaDome at different concentrations in BT-474 cells expressing HER2.

[0038] Figure 25 shows the results of flow cytometry analysis to determine the binding ability of the multispecific fusion protein [anti-ANXA1-anti-CD47] containing ReleaDome to each cell (CT-26 cells, MDA-MB-231 cells). Figures 25a and 25b show the results of concentration-dependent antigen [CD47] binding ability of the multispecific fusion protein [anti-ANXA1-anti-CD47] containing ReleaDome in CT-26 cells expressing mouse CD47. Figures 25c to 25f show the results of concentration-dependent antigen [CD47] binding ability of the multispecific fusion protein [anti-ANXA1-anti-CD47] containing ReleaDome in MDA-MB-231 cells expressing human CD47.

[0039] Figure 26 shows the results of flow cytometry analysis to determine the binding affinity of each tumor cell line (MDA-MB-231 cells, MDA-MB-453 cells) of a multispecific fusion protein [anti-ANXA1-anti-EGFR] including ReleaDome, which is an example of the present invention. Figures 26a to 26d show the results of determining the antigen [EGFR] binding affinity of each multispecific fusion protein [anti-ANXA1-anti-EGFR] including ReleaDome at different concentrations in MDA-MB-231 cells expressing human EGFR. Figure 26e shows the results of determining the antigen binding affinity of each fusion protein in MDA-MB-453 cells where human EGFR is hardly expressed.

[0040] Figure 27 shows the results of analyzing the transport ability (in vitro transcytosis) of the first domain (UNIT1) of ReleaDome, which shows binding affinity to ANXA1, through ELISA. Figure 27a shows the results of analyzing the transport ability of a single antibody containing each of the first domains (UNIT1) under co-culture conditions of EA.hy926 cells and MCF-7 cells. Figure 27b shows the results of analyzing the transport ability of a single antibody containing each of the first domains (UNIT1) under co-culture conditions of HUVEC cells and MCF-7 cells. Figure 27c shows the results of analyzing the transport ability of a single antibody and a multi-specific fusion protein under co-culture conditions of HUVEC cells and MCF-7 cells, respectively.

[0041] Figure 28 shows the results of analyzing the in vitro cytotoxicity of each multispecific fusion protein-drug conjugate containing ReleaDome according to treatment with a protease. Figure 28a shows the results of examining the cytotoxicity after treating NCI-N87 cells expressing HER2 with TD-0102-001.017 or (+)TD-0102-001.017 whose first domain is cleaved by a protease (MT-SP1 and uPA) at various concentrations. Figure 28b shows the results of examining the cytotoxicity after treating BT-474 cells expressing HER2 with TD-0102-001.017 or (+)TD-0102-001.017 whose first domain is cleaved by a protease (MT-SP1) at various concentrations. Figure 28c shows the results of examining cytotoxicity after treating SK-BR-3 cells expressing HER2 with TD-0102-001.017 or (+)TD-0102-001.017 whose first domain is cleaved by a protease (MT-SP1) at various concentrations. Figure 28d shows the results of examining cytotoxicity after treating JIMT-1 cells expressing HER2 with TD-0102-001.017 or (+)TD-0102-001.017 whose first domain is cleaved by a protease (MT-SP1) at various concentrations. Figure 28e shows the results of examining cytotoxicity after treating SK-OV-3 cells expressing HER2 with TD-0102-001.017 or (+)TD-0102-001.017 with the first domain cleaved by a protease (MT-SP1) at various concentrations.

[0042] Figure 29 shows the results of examining cytotoxicity after treating EGFR-expressing MDA-MB-231 cells with each multi-specific fusion protein-drug conjugate containing ReleaDome at different concentrations.

[0043] Figures 30a to 30j are examples of results of analyzing the thermal stability (differential scanning fluorimetry) of a multi-specific fusion protein containing ReleaDome.

[0044] Figure 31 shows the results of analyzing the stability in human plasma of each multi-specific fusion protein including ReleaDome through SDS-PAGE under reducing conditions. Figure 31a shows the results of confirming each multi-specific fusion protein conjugated with a fluorescent dye. Figures 31b to 31c show the results of analyzing the stability of each multi-specific fusion protein conjugated with a fluorescent dye by mixing it with human plasma and reacting it at 37°C on a daily basis.

[0045] Figure 32 shows the results of analyzing the pharmacokinetics (PK) of each multi-specific fusion protein including ReleaDome, which is an example of the present invention, using mice. Figure 32a shows the results of analyzing the pharmacokinetics of BA-0109-012 (4 mg / kg), Figure 32b shows the results of analyzing the pharmacokinetics of BA-0109-016 (4 mg / kg), and Figure 32c is a graph showing the results of analyzing the pharmacokinetics of BA-0109-018 (4 mg / kg).

[0046] Figure 33 shows the results of analyzing the distribution of a multi-specific fusion protein [anti-ANXA1-anti-HER2] including ReleaDome, which is an embodiment of the present invention, in cancer tissue using a tumor cell line transplantation tumor mouse model through tomographic optical imaging (IVIS®). Figure 33a shows the results of confirming the in vivo distribution through IVIS® 4 hours after administering each multi-specific fusion protein to an NCI-N87 (human gastric cancer cell) xenograft (CDX, Cell-derived xenograft) mouse model expressing HER2. Figure 33b shows the results of confirming the in vivo distribution through IVIS® 4 hours after administering each multi-specific fusion protein to a JIMT-1 human breast cancer CDX mouse model expressing HER2.

[0047] Figure 34 shows the results of analyzing the distribution in biological tissues of a multi-specific fusion protein (anti-ANXA1-anti-EGFR; TD-0109-003-CW) containing ReleaDome, which is an example of the present invention, in a CDX mouse model of JIMT-1 cells (human breast cancer cells) expressing EGFR using IVIS®. Figure 34a shows the results of confirming the distribution in vivo in mice at each time point through images after administration of TD-0109-003-CW, and Figure 34b shows the results of measuring the fluorescence intensity of major organs extracted from mice 72 hours after administration of TD-0109-003-CW.

[0048] Figure 35 shows the results of analyzing the in vivo toxicity (safety assessment) of a multi-specific fusion protein (anti-ANXA1-anti-CD47; BA-0107-005) containing ReleaDome, which is an example of the present invention.

[0049] Figure 36 shows the results of confirming the anticancer activity of each multispecific fusion protein [anti-ANXA1-anti-HER2]-drug conjugate including ReleaDome, which is an embodiment of the present invention, in a HER2-expressing NCI-N87 cell CDX mouse model. Figure 36a shows the results of measuring tumor size by day after administration of the test substance, and Figure 36b shows the results of measuring the weight of tumors excised from the mouse on the 62nd day after administration of the test substance.

[0050] Figure 37 shows the results of evaluating the anticancer activity of each multispecific fusion protein [anti-ANXA1-anti-EGFR]-drug conjugate including ReleaDome, which is an embodiment of the present invention, in a CDX mouse model of EGFR-expressing MDA-MB-231 cells (human triple-negative breast cancer). Figure 37a shows the results of measuring tumor size by date after test substance administration, and Figure 37b shows the results of measuring mouse body weight by date after test substance administration.

[0051] Figure 38 shows the results of measuring tumor size by date after administering each multispecific fusion protein [anti-ANXA1-anti-HER2]-drug conjugate (0.0175 umol / kg) or single antibody-drug conjugate (0.0338 umol / kg) including ReleaDome, which is an example of the present invention, to a JIMT-1 cell CDX mouse model expressing HER2. Figure 38a shows the results of measuring body weight of mice by date after administration of each test substance.

[0052] <Definition of Terms>

[0053] The term "antibody" as used herein refers to an immunoglobulin (Ig) molecule that immunologically reacts with a specific antigen, a protein molecule that acts as a receptor that specifically recognizes the antigen, and is a concept that encompasses both whole antibodies and antibody fragments. Antibodies are also referred to as immunoglobulins. Antibodies may refer to any one selected from IgG, IgE, IgM, IgD, and IgA, and may be subclasses of IgG, such as IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2. Furthermore, antibodies may be agonistic or antagonistic antibodies.

[0054] Specifically, the antibody or fragment thereof may include, but is not limited to, a monoclonal antibody, a polyclonal antibody, a single domain antibody, a single chain antibody, a multispecific antibody, a human antibody, a humanized antibody, a chimeric antibody, an intrabody, an Fv, a scFv, an Fv linked by a disulfide bond (di-scFv), a Fab fragment, an F(ab')2 fragment, a domain antibody, a minibody, a nanobody, a VHH (heavy chain variable domain of heavy chain antibody), an IgD antibody, an IgE antibody, an IgM antibody, an IgG1 antibody, an IgG2 antibody, an IgG3 antibody, an IgG4 antibody, an antibody constant region derivative, and any epitope binding fragment.

[0055] As used herein, the term "antigen" refers to a structure capable of selectively binding to an antibody. The target antigen may be a polypeptide, carbohydrate, nucleic acid, lipid, hapten, or other naturally occurring or synthetic compound. Specifically, the antigen may be a polypeptide or a protein present on or within a cell.

[0056] As used herein, the term "epitope" refers to an antigenic determinant and is a region on an antigen to which an antibody or polypeptide binds. A protein epitope may include amino acid residues directly involved in binding, as well as amino acid residues that are effectively blocked by a specific antigen-binding antibody or peptide. It is the simplest form or smallest structural region of a complex antigen molecule capable of binding to an antibody or receptor. Epitopes may be linear or structural / conformational.

[0057] As used herein, the term "Fv", "Fv fragment" or "Fv region" is a polypeptide comprising the variable region of heavy chain (VH) and the variable region of light chain (VL) of a single antibody.

[0058] As used herein, the term "single-chain Fv (scFv)" refers to an antibody fragment comprising the heavy chain variable region and the light chain variable region of an antibody within a single polypeptide chain.

[0059] As used herein, the term "Fab" or "Fab region" refers to the region of an antibody that binds to an antigen. A typical IgG generally comprises two Fab regions. Each Fab region typically consists of one variable region and one constant region of each heavy and light chain. Specifically, the variable and constant regions of the heavy chain in the Fab region are the VH and CH1 (Constant region of heavy chain 1) regions, and the variable and constant regions of the light chain in the Fab region are the VL and CL (Constant region of light chain) regions. The VH, CH1, VL, and CL of the Fab region can be arranged in various ways to impart antigen-binding ability according to the present disclosure, including a CrossMab Fab configuration in which the VH and VL are arranged in a permuted arrangement.

[0060] As used herein, the term "single domain antibody (sdAb)" refers to an antibody fragment composed of a single monomeric variable domain, also known as a nanobody (VHH, heavy chain variable domain of heavy chain antibody). It generally refers to an antibody that has antigen-binding activity comprising only one heavy chain variable region. While sdAbs derived from heavy chains are primarily used, single variable region fragments derived from light chains have also been reported to specifically bind to antigens.

[0061] As used herein, the term "heavy chain" refers to a polypeptide chain of about 50 kDa to about 70 kDa, wherein the N-terminal portion comprises a variable region of about 120 to 130 amino acids or more, and the C-terminal portion comprises a constant region. The constant region can be one of five types: alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ). Here, α, δ, and γ comprise about 450 amino acids, and μ and ε comprise about 550 amino acids. The constant domains (CH domains) of the heavy chain are numbered from the N-terminus to the C-terminus (e.g., CH1, CH2, CH3, etc.).

[0062] As used herein, the term "light chain" refers to a polypeptide chain of about 25 kDa, wherein the N-terminal portion comprises a variable region of about 100 to about 110 amino acids or more, and the C-terminal portion comprises a constant region. There are two types of light chain constant regions (CLs): kappa (κ) and lambda (λ). Any CL and CH1 regions of these antibody classes or variants thereof can be used in the present disclosure.

[0063] In an exemplary embodiment of the present invention, CH1 and CL may each comprise an amino acid sequence as shown in Table 1 below. The amino acid sequences below are sequentially listed from the N-terminus to the C-terminus.

[0064]

[0065]

[0066] The structural formula of the fusion protein structure described as one specific example of the present invention is described in Table 2 below.

[0067] Structural formulaStructural formula(I)1st structureStructural formula(II)2nd structureStructural formula(IV)4th structureStructural formula(V)5th structureStructural formula(I')6th structureStructural formula(IV')7th structureStructural formula(I'')8th structureStructural formula(IV'')9th structureStructural formula(I''')11th structureStructural formula(I'''')12th structureStructural formula(IV''')13th structureStructural formula(IV'''')14th structureStructural formula(VI)15th structureStructural formula(VII)16th structureStructural formula(VIII)3rd structure

[0068] The terms described in Tables 1 and 2 above are described in more detail below.

[0069] Fusion protein

[0070] One aspect of the present invention provides a fusion protein comprising a first domain comprising a site that specifically binds to a first target; and a second domain comprising a site that specifically binds to an immune modulator or a second target. In this case, the first target may be a protein overexpressed in vascular cells or basal cells at a disease site, and the second target may be a protein overexpressed or secreted on the surface of cancer cells or immune cells.

[0071] The first domain and the second domain may be connected via a linker, and the linker may be truncated or non-truncated. The fusion protein comprising the first domain and the second domain may further comprise an anticancer agent, and the anticancer agent and the fusion protein may be connected via a linker.

[0072] In addition, the fusion protein may additionally include an Fc region or a variant thereof and / or an antigen binding site. In this case, the fusion protein and the Fc region or a variant thereof or the antigen binding site may be linked via a linker. In addition, when the fusion protein and the antigen binding site are linked via a linker, the antigen binding site may include only the heavy chain variable region or the light chain variable region of the antibody.

[0073] In this novel structure, the first domain can suppress the activity of the second domain at the non-diseased site by shielding the second domain at the non-diseased site. Furthermore, the first domain can be cleaved at the diseased site by a disease-specific cleavage enzyme, thereby exposing the second domain, allowing it to be specifically activated at the diseased site, thereby efficiently exerting its efficacy.

[0074] The specific structure of the above fusion protein is described in more detail below.

[0075] Lilydom

[0076] One aspect of the present invention provides a fusion protein comprising a first domain comprising a site that specifically binds to a first target; and a second domain comprising a site that specifically binds to an immune modulator or a second target.

[0077] As used herein, the term "ReleaDome" refers to a dual-functional domain structure comprising the first domain and the second domain. The first domain and the second domain may be connected via a linker. Therefore, in the present invention, "ReleaDome" refers to a structure comprising the first domain, the second domain, and a linker. In the present invention, the ReleaDome may be the minimum unit of the fusion protein according to the present invention.

[0078] First domain

[0079] In the present invention, the first domain may include a site that specifically binds to the first target. The first domain may be described herein interchangeably with "first component," "A," or "outer domain." The first domain may shield the second domain at a non-diseased site, and may be cleaved at a diseased site to expose the second domain at the diseased site, thereby activating the second domain specifically at the diseased site.

[0080] The first target may be a protein overexpressed or overexposed in vascular cells or basal cells surrounding the disease site. Furthermore, the first target may be a protein overexpressed or overexposed in immune cells or diseased cells within the microenvironment of the disease site. The fusion protein according to the present invention, through the first domain, can position ReleaDome at the disease site in a location-specific and expression-specific manner.

[0081] Specifically, the first target may be Annexin A1 (ANXA1), Albumin, App2, B7H3, Cav-1, CD36, EphA2, EphA3, EphB1, FAPα (Fibroblast activation protein alpha), ICAM1, Integrins, KDR (VEGF2), TEM8, TSP-1, ED-B, Endothelin-1, Endoglin (CD105), VEGFR1 (FLT1), PDGFR, PSMA, VCAM-1, Myct1, etc. More specifically, it may be, but is not limited to, ANXA1, TEM8, Albumin, KDR (VEGFR2), PSMA, ICAM1, FAPα, B7H3, or Myct1.

[0082] The first domain may be any one selected from the group consisting of an antibody or a fragment thereof, a ligand or a fragment thereof, and a receptor or a fragment thereof for the first target, but is not limited thereto. The form of the first domain may be, for example, an antibody or a fragment thereof (e.g., scFv, Fv, VHH, Fab), a peptide, an aptamer, etc., and may be a form that includes part or all of a counterpart / ligand existing in nature of the target that can be an antigen of the first domain, but is not limited thereto. Preferably, it may be an antibody or a fragment thereof, or a peptide. The antibody and the fragment thereof are the same as described above. In one specific embodiment of the present invention, the antibody fragment may be a scFv or a single domain antibody.

[0083] Specifically, the first domain may be an antibody or a fragment thereof, or a peptide that binds to any one selected from the group consisting of ANXA1, TEM8, Albumin, KDR (VEGFR2), PSMA, ICAM1, FAPα, B7H3, and Myct1. Table 3 below shows the variable region polypeptide sequences or peptide sequences of the antibodies or fragments thereof described as the first domains of various fusion proteins described herein. Fusion proteins according to exemplary embodiments may include these variable region polypeptide or peptide sequences. The following amino acid sequences are sequentially listed from the N-terminus to the C-terminus.

[0084]

[0085]

[0086]

[0087]

[0088]

[0089]

[0090]

[0091]

[0092]

[0093]

[0094]

[0095]

[0096]

[0097] - ANXA1

[0098] More specifically, the first domain may be a peptide, antibody or fragment thereof that binds to ANXA1.

[0099] The term "Annexin A1" as used herein is also known as Lipocortin 1, Calpactin II, Chromobindin-9, Phospholipase A2 inhibitory protein, or p35. In general, ANXA1 is known to be involved in signal transduction between cells and its function is regulated according to calcium (Ca) concentration. In addition, ANXA1 is considered to be a factor affecting the growth and metastasis of cancer cells, and overexpression of ANXA1 has been reported in various cancers.

[0100] In the present invention, ANXA1 may be included without limitation as long as it is derived from mammals, including humans, primates such as monkeys, and rodents such as rats and mice. In addition, the ANXA1 protein may include both a native or mutant ANXA1 protein, but is not limited thereto. The "wild type" includes all proteins found in nature or nucleic acids encoding the same, and may be described interchangeably with wild type. The native ANXA1 protein generally refers to a polypeptide including the amino acid sequence of the native ANXA1 protein. The amino acid sequence and polynucleotide sequence for the ANXA1 protein can be obtained from known databases such as GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably includes the amino acid sequence of SEQ ID NO: 1616. In addition, as long as it has the same activity as the protein or has the same gene location encoding the ANXA1 on the chromosome, it may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted. Specifically, ANXA1 may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of SEQ ID NO: 1616.

[0101] In the present invention, the ANXA1 protein variant refers to a fragment of ANXA1 in which a portion of the N-terminus is deleted. Specifically, the ANXA1 protein variant may be in a form that includes amino acids in which the 1st to 26th amino acids from the N-terminus of a protein having an amino acid sequence of SEQ ID NO: 1616 are consecutively deleted. Specifically, it may include an amino acid sequence of SEQ ID NO: 1617. In addition, as long as it has the same activity as the variant or the gene position encoding the ANXA1 on the chromosome is the same, it may be composed of a sequence in which one or more amino acids are added, deleted, or substituted. Specifically, it may include or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of SEQ ID NO: 1617.

[0102] In one specific embodiment of the present invention, the first domain may be a peptide fragment comprising the amino acid sequence of SEQ ID NO: 145.

[0103] In one specific embodiment of the present invention, the first domain may be an antibody or a fragment thereof. The antibody or fragment thereof comprises an HCDR1 comprising any one amino acid sequence selected from the group consisting of SEQ ID NOs: 277, 284, 291, 298, 304, 310, 320, 326, 332, 336, 338, 344, 355, 370, 382, ​​392, 396, 398, 405, 411, 417, 420, 427, 432, 436, 441, 444, 449, 456, 460, 465, 470, 475, 478 and 489, SEQ ID NOs: 279, 286, 293, 300, 305, 322, 328, 334, HCDR2 comprising any one amino acid sequence selected from the group consisting of 340, 346, 351, 357, 365, 372, 378, 386, 394, 400, 406, 413, 422, 429, 434, 442, 446, 451, 458, 462, 467, 472, 480, 486, 491, 494, 495, 496, 497 and 498, and SEQ ID NOs: 281, 288, 295, 302, 307, 312, 317, 324, 330, 342, 348, 353, 359, 361, 362, A heavy chain variable region comprising an HCDR3 comprising any one amino acid sequence selected from the group consisting of 374, 380, 388, 402, 408, 415, 424, 453, and 492;and LCDR1 comprising any one amino acid sequence selected from the group consisting of SEQ ID NOs: 500, 512, 525, 531, 538, 551, 557, 564, 570, 575, 580, 592, 595, 599, 603, 608, 616, 621, 625, 631, 636, 645, 651, 654, 659, 667, 670, 675, 678, 684, 690, 698, 704, 708, 711 and 714, SEQ ID NOs: 502, 508, 514, 523, 527, 533, 540, 547, LCDR2 comprising any one amino acid sequence selected from the group consisting of 553, 559, 566, 582, 587, 609, 617, 632, 639, 647, 663, 680, 685 and 716, and SEQ ID NOs: 504, 510, 516, 529, 535, 542, 544, 549, 555, 561, 568, 572, 578, 584, 586, 588, 590, 597, 602, 606, 611, 613, 619, 624, 627, 630, 634, 641, 643, 649, 653, A light chain variable region comprising an LCDR3 comprising any one amino acid sequence selected from the group consisting of 662, 665, 669, 673, 674, 682, 687, 689, 693, 696, 697, 718 and 719.;

[0104] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 277, HCDR2 comprising the amino acid sequence of SEQ ID NO: 279, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 281, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 500, LCDR2 comprising the amino acid sequence of SEQ ID NO: 502, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 504. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 1, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 2 (A1).

[0105] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 284, HCDR2 comprising the amino acid sequence of SEQ ID NO: 286, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 288, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 500, LCDR2 comprising the amino acid sequence of SEQ ID NO: 508, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 510. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 3, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 4 (A2).

[0106] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 516. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 5, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 6 (A3).

[0107] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 298, HCDR2 comprising the amino acid sequence of SEQ ID NO: 300, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 302, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 500, LCDR2 comprising the amino acid sequence of SEQ ID NO: 502, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 504. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 7, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 8 (A4).

[0108] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 304, HCDR2 comprising the amino acid sequence of SEQ ID NO: 305, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 307, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 500, LCDR2 comprising the amino acid sequence of SEQ ID NO: 523, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 510. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 9, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 10 (A5).

[0109] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 310, HCDR2 comprising the amino acid sequence of SEQ ID NO: 286, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 312, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 525, LCDR2 comprising the amino acid sequence of SEQ ID NO: 527, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 529. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 11, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 12 (A6).

[0110] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 317, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 531, LCDR2 comprising the amino acid sequence of SEQ ID NO: 533, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 535. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 13, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 14 (A7).

[0111] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 320, HCDR2 comprising the amino acid sequence of SEQ ID NO: 322, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 324, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 538, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 542. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 15, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 16 (A8).

[0112] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 326, HCDR2 comprising the amino acid sequence of SEQ ID NO: 328, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 330, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 500, LCDR2 comprising the amino acid sequence of SEQ ID NO: 502, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 544. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 17, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 18 (A9).

[0113] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 332, HCDR2 comprising the amino acid sequence of SEQ ID NO: 334, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 547, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 549. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 19, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 20 (A10).

[0114] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 336, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 551, LCDR2 comprising the amino acid sequence of SEQ ID NO: 553, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 555. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 21, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 22 (A11).

[0115] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 338, HCDR2 comprising the amino acid sequence of SEQ ID NO: 340, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 342, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 557, LCDR2 comprising the amino acid sequence of SEQ ID NO: 559, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 561. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 23, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 24 (A12).

[0116] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 344, HCDR2 comprising the amino acid sequence of SEQ ID NO: 346, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 348, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 564, LCDR2 comprising the amino acid sequence of SEQ ID NO: 566, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 568. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 25, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 26 (A13).

[0117] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 351, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 353, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 570, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 555. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 27, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 28 (A14).

[0118] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 355, HCDR2 comprising the amino acid sequence of SEQ ID NO: 357, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 572. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 29, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 30 (A15).

[0119] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 361, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 575, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 578. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 31, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 32 (A16).

[0120] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 362, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 580, LCDR2 comprising the amino acid sequence of SEQ ID NO: 582, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 584. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 33, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 34 (A17).

[0121] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 365, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 586. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 35, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 36 (A18).

[0122] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 587, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 588. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 37, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 38 (A19).

[0123] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 370, HCDR2 comprising the amino acid sequence of SEQ ID NO: 372, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 374, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 564, LCDR2 comprising the amino acid sequence of SEQ ID NO: 582, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 590. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 39, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 40 (A20).

[0124] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 592, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 572. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 41, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 42 (A21).

[0125] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 355, HCDR2 comprising the amino acid sequence of SEQ ID NO: 378, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 380, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 595, LCDR2 comprising the amino acid sequence of SEQ ID NO: 582, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 597. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 43, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 44 (A22).

[0126] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 382, ​​HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 572. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 45, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 46 (A23).

[0127] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 286, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 599, LCDR2 comprising the amino acid sequence of SEQ ID NO: 582, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 602. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 47, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 48 (A24).

[0128] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 386, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 388, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 603, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 606. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 49, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 50 (A25).

[0129] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 608, LCDR2 comprising the amino acid sequence of SEQ ID NO: 609, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 611. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 51, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 52 (A26).

[0130] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 613. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 53, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 54 (A27).

[0131] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 392, HCDR2 comprising the amino acid sequence of SEQ ID NO: 394, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 616, LCDR2 comprising the amino acid sequence of SEQ ID NO: 617, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 619. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 55, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 56 (A28).

[0132] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 621, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 602. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 57, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 58 (A29).

[0133] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 608, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 624. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 59, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 60 (A30).

[0134] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 396, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 625, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 627. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 61, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 62 (A31).

[0135] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 398, HCDR2 comprising the amino acid sequence of SEQ ID NO: 400, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 402, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 625, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 630. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 63, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 64 (A32).

[0136] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 405, HCDR2 comprising the amino acid sequence of SEQ ID NO: 406, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 408, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 631, LCDR2 comprising the amino acid sequence of SEQ ID NO: 632, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 634. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 65, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 66 (A33).

[0137] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 411, HCDR2 comprising the amino acid sequence of SEQ ID NO: 413, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 415, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 636, LCDR2 comprising the amino acid sequence of SEQ ID NO: 639, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 641. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 67, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 68 (A34).

[0138] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, HCDR2 comprising the amino acid sequence of SEQ ID NO: 351, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 603, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 643. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 69, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 70 (A35).

[0139] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 420, HCDR2 comprising the amino acid sequence of SEQ ID NO: 422, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 424, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 645, LCDR2 comprising the amino acid sequence of SEQ ID NO: 647, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 649. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 71, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 72 (A36).

[0140] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 320, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 651, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 653. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 73, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 74 (A37).

[0141] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 427, HCDR2 comprising the amino acid sequence of SEQ ID NO: 429, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 654, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 602. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 75, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 76 (A38).

[0142] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 432, HCDR2 comprising the amino acid sequence of SEQ ID NO: 434, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 624. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 77, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 78 (A39).

[0143] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 436, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 659, LCDR2 comprising the amino acid sequence of SEQ ID NO: 582, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 662. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 79, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 80 (A40).

[0144] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, HCDR2 comprising the amino acid sequence of SEQ ID NO: 351, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 531, LCDR2 comprising the amino acid sequence of SEQ ID NO: 663, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 665. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 81, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 82 (A41).

[0145] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 667, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 669. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 83, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 84 (A42).

[0146] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 320, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 670, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 673. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 85, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 86 (A43).

[0147] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 674. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 87, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 88 (A44).

[0148] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 441, HCDR2 comprising the amino acid sequence of SEQ ID NO: 442, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 675, LCDR2 comprising the amino acid sequence of SEQ ID NO: 553, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 643. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 89, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 90 (A45).

[0149] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 444, HCDR2 comprising the amino acid sequence of SEQ ID NO: 446, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 678, LCDR2 comprising the amino acid sequence of SEQ ID NO: 680, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 682. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 91, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 92 (A46).

[0150] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 449, HCDR2 comprising the amino acid sequence of SEQ ID NO: 451, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 453, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 684, LCDR2 comprising the amino acid sequence of SEQ ID NO: 685, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 687. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 93, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 94 (A47).

[0151] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 456, HCDR2 comprising the amino acid sequence of SEQ ID NO: 458, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 689. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 95, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 96 (A48).

[0152] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 460, HCDR2 comprising the amino acid sequence of SEQ ID NO: 462, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 690, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 693. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 97, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 98 (A49).

[0153] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 465, HCDR2 comprising the amino acid sequence of SEQ ID NO: 467, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 696. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 99, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 100 (A50).

[0154] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 470, HCDR2 comprising the amino acid sequence of SEQ ID NO: 472, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 609, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 697. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 101, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 102 (A51).

[0155] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 475, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 698, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 627. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 103, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 104 (A52).

[0156] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 478, HCDR2 comprising the amino acid sequence of SEQ ID NO: 480, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 696. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 105, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 106 (A53).

[0157] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 704, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 602. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 107, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 108 (A54).

[0158] In one specific embodiment, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, HCDR2 comprising the amino acid sequence of SEQ ID NO: 351, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 708, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 602. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 109, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 110 (A55 / A82). In addition, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 118, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 110 (A69 / A77 / A77-1).

[0159] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, HCDR2 comprising the amino acid sequence of SEQ ID NO: 351, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 711, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 602. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 111, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 112 (A56).

[0160] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 486, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 678, LCDR2 comprising the amino acid sequence of SEQ ID NO: 680, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 682. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 113, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 114 (A57).

[0161] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 489, HCDR2 comprising the amino acid sequence of SEQ ID NO: 491, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 492, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 714, LCDR2 comprising the amino acid sequence of SEQ ID NO: 716, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 718. In this case, the light chain variable region may include the amino acid sequence of SEQ ID NO: 115, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 116 (A58).

[0162] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, HCDR2 comprising the amino acid sequence of SEQ ID NO: 351, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 609, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 719. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 109, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 117 (A59).

[0163] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, HCDR2 comprising the amino acid sequence of SEQ ID NO: 351, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 609, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 719. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 118, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 117 (A60).

[0164] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, HCDR2 comprising the amino acid sequence of SEQ ID NO: 494, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 708, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 602. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 119, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 110 (A61). In addition, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 120, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 110 (A62).

[0165] In one specific embodiment, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, HCDR2 comprising the amino acid sequence of SEQ ID NO: 495, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 708, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 602. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 121, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 110 (A63).

[0166] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, HCDR2 comprising the amino acid sequence of SEQ ID NO: 496, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 708, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 602. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 122, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 110 (A64). In addition, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 123, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 110 (A65).

[0167] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, HCDR2 comprising the amino acid sequence of SEQ ID NO: 497, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 708, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 602. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 124, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 110 (A66).

[0168] In one specific example, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, HCDR2 comprising the amino acid sequence of SEQ ID NO: 498, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 708, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 602. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 125, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 110 (A67). In addition, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 126, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 110 (A68).

[0169] In the present invention, the antibody fragment may be a single domain antibody. In this case, the antibody fragment may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295. The heavy chain variable region may include the amino acid sequence of SEQ ID NO: 142 (A78).

[0170] - TEM8

[0171] More specifically, the first domain may be an antibody or fragment thereof that binds to TEM8.

[0172] The term "TEM8" as used herein is also known as Anthrax toxin receptor 1 (ANTXR1), a type 1 transmembrane protein. TEM8 is known to be overexpressed in endothelial cells or tumor cells surrounding various tumors, including colon cancer.

[0173] In the present invention, TEM8 may be included without limitation as long as it is derived from mammals, including primates such as humans and monkeys, and rodents such as rats and mice. In addition, the TEM8 protein may include both a native or mutant TEM8 protein, but is not limited thereto. The native TEM8 protein generally refers to a polypeptide including the amino acid sequence of the native TEM8 protein. The amino acid sequence and polynucleotide sequence for the TEM8 protein can be obtained from known databases such as the GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably includes the amino acid sequence of SEQ ID NO: 177.

[0174] In addition, it may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted, as long as it has the same activity as the protein or the gene location encoding the TEM8 on the chromosome is the same. Specifically, TEM8 may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of SEQ ID NO: 177.

[0175] In one specific embodiment, the antibody or fragment thereof may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 721, HCDR2 comprising the amino acid sequence of SEQ ID NO: 723, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 725, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 763, LCDR2 comprising the amino acid sequence of SEQ ID NO: 765, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 767. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 127, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 128 (A70 / A81).

[0176] - Albumin

[0177] More specifically, the first domain may be an antibody or a fragment thereof that binds to albumin.

[0178] The term "albumin" used in this specification is a simple protein that exists in nature and is one of the proteins that constitute the basic substance of cells, and has a molecular weight of 65-70 kDa.

[0179] In the present invention, albumin may be included without limitation as long as it is derived from mammals, including primates such as humans and monkeys, and rodents such as rats and mice. In addition, the albumin protein may include both native and mutant albumin proteins, but is not limited thereto. The native albumin protein generally refers to a polypeptide comprising the amino acid sequence of the native albumin protein. The amino acid sequence and polynucleotide sequence for the albumin protein can be obtained from known databases such as GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably may include the amino acid sequence of SEQ ID NO: 178.

[0180] Additionally, it may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted, as long as it has the same activity as the protein or has the same gene location on the chromosome that encodes the albumin. Specifically, the albumin may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of SEQ ID NO: 178.

[0181] In one specific embodiment of the present invention, the first domain may include a heavy chain variable region comprising HCDR1 comprising an amino acid sequence of SEQ ID NO: 727, HCDR2 comprising an amino acid sequence of SEQ ID NO: 729, and HCDR3 comprising an amino acid sequence of SEQ ID NO: 731, and a light chain variable region comprising LCDR1 comprising an amino acid sequence of SEQ ID NO: 770, LCDR2 comprising an amino acid sequence of SEQ ID NO: 772, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 774. In this case, the heavy chain variable region may include an amino acid sequence of SEQ ID NO: 129, and the light chain variable region may include an amino acid sequence of SEQ ID NO: 130 (A71).

[0182] - KDR (VEGFR2)

[0183] More specifically, the first domain may be an antibody or fragment thereof that binds to KDR (VEGFR2, Vascular endothelial growth factor receptor 2).

[0184] The term "KDR" as used herein is also known as vascular endothelial growth factor receptor 2 (VEGFR2; KDR / Flk-1). Vascular endothelial growth factor (VEGF) is a heparin-binding glycoprotein with a molecular weight of 34–42 kDa and is a representative growth factor that promotes endothelial cell migration. VEGF is known to have five isoforms (VEGF-A, -B, -C, -D, and F) and each is known to react with its own receptor. Known receptors include VEGFR1 (Flt-1; Fms-like-tyrosine kinase-1), VEGFR2 (flk-1 / KDR), VEGFR3, PDGF (Platelet-derived growth factor) receptor, and NP-1 (Neuropilin-1). VEGFR1 binds to VEGF-A, VEGF-B, or PDGF, VEGFR2 binds to VEGF-A, VEGF-C, VEGF-D, or VEGF-E, and VEGFR3 binds to VEGF-C or VEGF-D. Each of the above VEGF receptors is activated through autophosphorylation with the receptor dimer when VEGF binds. Recent reports have shown that VEGF-induced angiogenesis is mediated through the activation of Flt-1 and KDR expressed in endothelial cells. In particular, KDR has been shown to play an important role in cell growth, survival, and migration.

[0185] In the present invention, KDR may be included without limitation as long as it is derived from mammals, including primates such as humans and monkeys, and rodents such as rats and mice. In addition, the KDR protein may include both a native or mutant KDR protein, but is not limited thereto. The native KDR protein generally refers to a polypeptide comprising the amino acid sequence of the native KDR protein. The amino acid sequence and polynucleotide sequence for the KDR protein can be obtained from known databases such as GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably may include the amino acid sequence of SEQ ID NO: 179.

[0186] Additionally, the protein may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted, as long as it has the same activity as the protein or has the same gene location encoding the KDR on the chromosome. Specifically, the KDR protein may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of SEQ ID NO: 179.

[0187] In one specific embodiment of the present invention, the first domain may include a heavy chain variable region comprising HCDR1 comprising an amino acid sequence of SEQ ID NO: 733, HCDR2 comprising an amino acid sequence of SEQ ID NO: 735, and HCDR3 comprising an amino acid sequence of SEQ ID NO: 736, and a light chain variable region comprising LCDR1 comprising an amino acid sequence of SEQ ID NO: 775, LCDR2 comprising an amino acid sequence of SEQ ID NO: 566, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 778. In this case, the heavy chain variable region may include an amino acid sequence of SEQ ID NO: 131, and the light chain variable region may include an amino acid sequence of SEQ ID NO: 132 (A72).

[0188] - PSMA

[0189] More specifically, the first domain may be an antibody or a fragment thereof that binds to PSMA (Prostate specific membrane antigen).

[0190] As used herein, the term "PSMA" refers to a glycoprotein enzyme synthesized in the epithelial cells of the prostate. Because it is rarely produced in tissues other than the prostate, it serves as a useful tumor marker for screening and monitoring prostate cancer.

[0191] In the present invention, PSMA may be included without limitation as long as it is derived from mammals, including primates such as humans and monkeys, and rodents such as rats and mice. In addition, the PSMA protein may include both a native or mutant PSMA protein, but is not limited thereto. The native PSMA protein generally refers to a polypeptide comprising the amino acid sequence of the native PSMA protein. The amino acid sequence and polynucleotide sequence for the PSMA protein can be obtained from known databases such as GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably may include an amino acid sequence of 180.

[0192] Additionally, the protein may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted, as long as it has the same activity as the protein or has the same gene location on the chromosome that encodes the PSMA protein. Specifically, the PSMA protein may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of 180.

[0193] In one specific embodiment of the present invention, the first domain may include a heavy chain variable region comprising HCDR1 comprising an amino acid sequence of SEQ ID NO: 738, HCDR2 comprising an amino acid sequence of SEQ ID NO: 740, and HCDR3 comprising an amino acid sequence of SEQ ID NO: 742, and a light chain variable region comprising LCDR1 comprising an amino acid sequence of SEQ ID NO: 781, LCDR2 comprising an amino acid sequence of SEQ ID NO: 559, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 784. In this case, the heavy chain variable region may include an amino acid sequence of SEQ ID NO: 133, and the light chain variable region may include an amino acid sequence of SEQ ID NO: 134 (A73).

[0194] - ICAM1

[0195] More specifically, the first domain may be an antibody or a fragment thereof that binds to ICAM1 (Intercellular adhesion molecule 1).

[0196] As used herein, the term "ICAM1," also known as CD54, is a cell surface adhesion molecule belonging to the immunoglobulin superfamily. ICAM1 is a cell surface glycoprotein of approximately 90 kDa and, like the HLA-DR antigen, is distributed on most nucleated cells. It is known to be involved in cell-to-cell endocytosis by forming ligand bonds with the lymphocyte surface contact molecule LFA-1 (Lymphocyte function associated antigen-1).

[0197] In the present invention, ICAM1 may be included without limitation as long as it is derived from mammals, including primates such as humans and monkeys, and rodents such as rats and mice. In addition, the ICAM1 protein may include both native and mutant ICAM1 proteins, but is not limited thereto. The native ICAM1 protein generally refers to a polypeptide comprising the amino acid sequence of the native ICAM1 protein. The amino acid sequence and polynucleotide sequence for the ICAM1 protein can be obtained from known databases such as the GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably may include an amino acid sequence of 181.

[0198] In addition, as long as it has the same activity as the protein or has the same gene location encoding the ICAM1 protein on the chromosome, it may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted. Specifically, the ICAM1 protein may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of 181.

[0199] In one specific embodiment of the present invention, the first domain may include a heavy chain variable region comprising HCDR1 comprising an amino acid sequence of SEQ ID NO: 745, HCDR2 comprising an amino acid sequence of SEQ ID NO: 747, and HCDR3 comprising an amino acid sequence of SEQ ID NO: 749, and a light chain variable region comprising LCDR1 comprising an amino acid sequence of SEQ ID NO: 786, LCDR2 comprising an amino acid sequence of SEQ ID NO: 788, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 790. In this case, the heavy chain variable region may include an amino acid sequence of SEQ ID NO: 135, and the light chain variable region may include an amino acid sequence of SEQ ID NO: 136 (A74).

[0200] - FAPα

[0201] More specifically, the first domain may be an antibody or a fragment thereof that binds to FAPα (Fibroblast activation protein α).

[0202] As used herein, the term "FAPα" is a transmembrane protein of approximately 170 kDa, which exists in both an intracellular domain and a soluble form lacking the transmembrane domain. While generally absent from normal adult tissues, it is known to be overexpressed on reactive stromal fibroblasts in more than 90% of all primary and metastatic epithelial tumors, including those of the lung, colon, bladder, ovary, and breast. Furthermore, it is expressed not only in stromal fibroblasts but also in some types of malignant cells of epithelial origin, and FAP expression is known to correlate with the malignant phenotype.

[0203] In the present invention, FAPα may be included without limitation as long as it is derived from mammals, including primates such as humans and monkeys, and rodents such as rats and mice. In addition, the FAPα protein may include both a native or mutant FAPα protein, but is not limited thereto. The native FAPα protein generally refers to a polypeptide comprising the amino acid sequence of the native FAPα protein. The amino acid sequence and polynucleotide sequence for the FAPα protein can be obtained from known databases such as the GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably may include an amino acid sequence of 182.

[0204] Additionally, the protein may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted, as long as it has the same activity as the protein or has the same gene location on the chromosome that encodes the FAPα protein. Specifically, the FAPα protein may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of 182.

[0205] In one specific embodiment of the present invention, the first domain may include a heavy chain variable region comprising HCDR1 comprising an amino acid sequence of SEQ ID NO: 751, HCDR2 comprising an amino acid sequence of SEQ ID NO: 753, and HCDR3 comprising an amino acid sequence of SEQ ID NO: 755, and a light chain variable region comprising LCDR1 comprising an amino acid sequence of SEQ ID NO: 792, LCDR2 comprising an amino acid sequence of SEQ ID NO: 794, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 796. In this case, the heavy chain variable region may include an amino acid sequence of SEQ ID NO: 137, and the light chain variable region may include an amino acid sequence of SEQ ID NO: 138 (A75).

[0206] - B7H3

[0207] More specifically, the first domain may be an antibody or fragment thereof that binds to B7H3 (B7 Homologue 3).

[0208] As used herein, the term "B7H3," also known as CD276 (Cluster of Differentiation 276), refers to an immune checkpoint inhibitor molecule. While highly restricted in normal cells, it is significantly increased in primary and metastatic tumors, as well as in tumor vasculature. Furthermore, it is expressed in numerous cell types, including differentiated tumor cells, tumor-initiating cells, and cancer stem cells, and its expression is known to be strongly correlated with poor prognosis in some tumor types.

[0209] In the present invention, B7H3 may be included without limitation as long as it is derived from mammals, including primates such as humans and monkeys, and rodents such as rats and mice. In addition, the B7H3 protein may include both a native or mutant B7H3 protein, but is not limited thereto. The native B7H3 protein generally refers to a polypeptide comprising the amino acid sequence of the native B7H3 protein. The amino acid sequence and polynucleotide sequence for the B7H3 protein can be obtained from known databases such as the GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably may include an amino acid sequence of 183.

[0210] Additionally, as long as it has the same activity as the protein or has the same gene location encoding the B7H3 protein on the chromosome, it may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted. Specifically, the B7H3 protein may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of 183.

[0211] In one specific embodiment of the present invention, the first domain may include a heavy chain variable region comprising HCDR1 comprising an amino acid sequence of SEQ ID NO: 757, HCDR2 comprising an amino acid sequence of SEQ ID NO: 759, and HCDR3 comprising an amino acid sequence of SEQ ID NO: 761, and a light chain variable region comprising LCDR1 comprising an amino acid sequence of SEQ ID NO: 580, LCDR2 comprising an amino acid sequence of SEQ ID NO: 566, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 799. In this case, the heavy chain variable region may include an amino acid sequence of SEQ ID NO: 139, and the light chain variable region may include an amino acid sequence of SEQ ID NO: 140 (A76).

[0212] -Myct1

[0213] More specifically, the first domain may be an antibody or a fragment thereof that binds to Myct1 (myc target 1).

[0214] The term "Myct1" used herein refers to a target gene of c-Myc, also known as MTLC. c-Myc is the most frequently activated oncoprotein during carcinogenesis, and regulates the expression of genes involved in cell cycle, apoptosis, and cellular transformation. Myct1 is transcriptionally promoted by c-Myc, and its abnormal expression has been reported to be associated with cancer occurrence, progression, metastasis, and prognosis. In particular, it is known as a key protein that regulates angiogenesis and immunity within the tumor microenvironment (TME). In addition, in blood-related diseases, it is known to play an important role in the self-renewal and differentiation of hematopoietic stem cells (HSCs), as well as the proliferation and apoptosis of acute myeloid leukemia (AML) cells.

[0215] In the present invention, Myct1 may be included without limitation as long as it is derived from mammals, including primates such as humans and monkeys, and rodents such as rats and mice. In addition, the Myct1 protein may include both native and mutant Myct1 proteins, but is not limited thereto. The native B7H3 protein generally refers to a polypeptide comprising the amino acid sequence of the native Myct1 protein. The amino acid sequence and polynucleotide sequence for the Myct1 protein can be obtained from known databases such as the GenBank of the National Center for Biotechnology Information (NCBI) in the United States.

[0216] Additionally, it may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted, as long as it has the same activity as the protein or has the same gene location on the chromosome that encodes the Myct1 protein.

[0217] In the present invention, the antibody or fragment thereof, peptide, or protein binding site forming the first domain is named “UNIT1.”

[0218] Second domain

[0219] In the present invention, the second domain may include a site that specifically binds to an immune modulator or a second target. The second domain may be described herein interchangeably with "third component," "C," or "inner domain."

[0220] In the present invention, the binding ability of the second domain of ReleaDome to the target protein can be shielded through the first domain at a non-disease site, and transferred to the target site of the first domain in an inactive state.

[0221] As used herein, the term "immunomodulator" refers to a regulatory substance that plays a key role in regulating the immune system at the site of disease. Specifically, the immunomodulator may be a cytokine, chemokine, or a fragment thereof. Furthermore, the immunomodulator may be a molecule that binds to the cytokine, chemokine, or a fragment thereof.

[0222] In the present invention, the second target may be a protein overexpressed, specifically expressed, or activated in target cells at the site of the disease. The target cells may be tumor cells, disease-related stromal cells, or immune cells.

[0223] In the present invention, the fusion protein can exhibit pharmacological activity against a disease by binding to a disease-specific target protein through the second domain.

[0224] 구체적으로, 상기 제2 표적은 4-1-BB, 5T4, ACE, ADAM9, ADRB3, AKAP4, ALK, ANXA1, ANXA2, ANXA5, ASCT2, AXL, B7H3, B7H4, BCL2, BCMA, CA125, CA6, CA9, CCL11, CCR5, CEN106, CD2, CD4, CD5, CD15, CD16, CD19, CD20, CD22, CD23, CD24, CD25, CD27, CD30, CD33, CD37, CD38, CD40, CD41, CD44, CD44v6, CD46, CD48, CD51, CD52, CD54, CD56, CD62, CD70, CD71, CD72, CD73, CD74, CD79, CD80, CD90, CD97, CD99, CD105, CD117, CD123, CD133, CD135, CD138, CD142, CD163, CD171, CD, CD242, CD319, CD352, CDCP1, CEA, CLDN3, CLDN6, CLDN9, CLDN18.2, CXCR2, CXCR4, CXCR5, CFC1B, CEACAM5, CEACAM6, c-Met, c-Kit, CTLA4, DLL3, DLK-1, DR5, DPEP3, Dectin-2, ED-B, EGFR, Endothelin-1, Endoglin, ENPP3, EpCAM, EphA2, ETA, FAPα, FGFR1, FGFR2, FGFR3, FRα, GD2, GD3, GloboH, GPC3, GPR20, GPRC5D, HER2, HER3, ICAM1, ICAM2, ICOS-L, IGF-1R, KAAG-1, L1CAM, LCK, LGR5, LIV-1, LRRC15, LY6E, MAGE, Mesothelin, MUC1, MUC16, MMP-14, NCAM1, Nectin-4, Notch3, NaPi2b, PDGFR, PD-L1, PD-L2, PSCA, PSMA, PTK7, RAGE-1, RANKL, RNF43, RON, ROR1, ROR2, SAP-1, SLAMF7, SLC44A4, Survivin, SAIL, STn, SSEA-4, SEZ6, SLC44A4, SLTRK6, TEM1, TEM8, TM4SF1, TNFLYPD3, TF, TIE2, TIM-1, TRAC, TROP-2, VEGFR1(FLT1), VEGFR2(KDR), VCAM-1, VSIG4, VISTA MEDICINE XAGE-1 있으나, snowflakes.

[0225] The form of the second domain may be, for example, an antibody or a fragment thereof (e.g., Fv, scFv, VHH, Fab), a peptide, an aptamer, etc., or may include part or all of a counterpart / ligand existing in nature of the target that can be the antigen of the second domain, but is not limited thereto. Preferably, it may be an antibody or a fragment thereof. The antibody and the fragment thereof are the same as described above. In one specific embodiment of the present invention, the fragment of the antibody may be a scFv or a single domain antibody.

[0226] Specifically, the second domain may be an antibody or fragment thereof that binds to any one selected from the group consisting of HER2, CD47, EGFR, CTLA4, B7H3, ANXA1, DLR1, CD3, CLDN18.2, TEM8, FAPα, EpCAM, PD-1, PD-L1, and Amyloid-beta (Aβ) protofibrils. Table 4 below shows the variable region polypeptide sequences of antibodies or fragments thereof described as the second domains of various fusion proteins described herein. Fusion proteins according to exemplary embodiments may include these variable region polypeptides.

[0227]

[0228]

[0229]

[0230]

[0231]

[0232]

[0233] - HER2

[0234] More specifically, the second domain may be an antibody or fragment thereof that binds to HER2.

[0235] As used herein, the term "HER2", also known as ErbB-2 or CD340, refers to a protein belonging to the ErbB (HER / EGFR / ERBB) family. The ErbB family consists of four types of binding receptor tyrosine kinase subfamilies: ErbB-1 (HER1, EGFR), ErbB-2 (HER2 / neu), ErbB-3 (HER3), and ErbB-4 (HER4). All four types of receptors comprise an extracellular ligand-binding domain, a transmembrane domain, and an intracellular domain. The intracellular domain interacts with a number of signaling molecules in a ligand-dependent or -independent manner. In particular, the ligand for HER2 is not yet known. HER2 is known to function by forming a heterodimer with one of the other three types of receptors. The ErbB family of receptors is known to be involved in regulating cell adhesion, migration, and differentiation in addition to cell proliferation and survival. In particular, HER2 is known to be the most potent oncoprotein in breast cancer. When HER2 is at normal levels, it participates in the growth and development of normal mammary tissue. However, when HER2 is abnormally overexpressed or amplified, normal cell regulation is disrupted, leading to the formation of aggressive cancer cells in the mammary tissue. HER2 overexpression occurs in approximately 20-30% of invasive breast cancers, and overexpression is known to be associated with a poor prognosis of breast cancer, with a higher malignancy rate and more aggressive cancer.

[0236] In the present invention, HER2 may be included without limitation as long as it is derived from mammals, including humans, primates such as monkeys, and rodents such as rats and mice. In addition, the HER2 protein may include both a native or mutant HER2 protein, but is not limited thereto. The native HER2 protein generally refers to a polypeptide comprising the amino acid sequence of the native HER2 protein. The amino acid sequence and polynucleotide sequence for the HER2 protein can be obtained from known databases such as the GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably may include the amino acid sequence of SEQ ID NO: 148.

[0237] Additionally, it may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted, as long as it has the same activity as the protein or has the same gene location encoding the HER2 on the chromosome. Specifically, HER2 may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of SEQ ID NO: 148.

[0238] In one specific embodiment of the present invention, the second domain may include a heavy chain variable region and a light chain variable region of Trastuzumab, an anti-HER2 antibody. Specifically, the second domain may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 802, HCDR2 comprising the amino acid sequence of SEQ ID NO: 804, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 806, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 913, LCDR2 comprising the amino acid sequence of SEQ ID NO: 680, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 915. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 184, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 185 (C1). Alternatively, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 186, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 187 (C2).

[0239] - CD47

[0240] More specifically, the second domain may be an antibody or fragment thereof that binds to CD47.

[0241] As used herein, the term "CD47" refers to a member of the immunoglobulin superfamily, also known as integrin-associated protein (IAP). CD47 is a cell surface immunoglobulin whose ligand interacts with SIRPα, which is primarily expressed on macrophages and dendritic cells, thereby inhibiting their phagocytosis. CD47 is known to be overexpressed in various tumors, but overexpression in normal tissues has also been reported.

[0242] In the present invention, CD47 may be included without limitation as long as it is derived from mammals, including primates such as humans and monkeys, and rodents such as rats and mice. In addition, the CD47 protein may include both native and mutant CD47 proteins, but is not limited thereto. The native CD47 protein generally refers to a polypeptide comprising the amino acid sequence of the native CD47 protein. The amino acid sequence and polynucleotide sequence for the CD47 protein can be obtained from known databases such as the GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably may include the amino acid sequence of SEQ ID NO: 149.

[0243] Additionally, it may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted, as long as it has the same activity as the protein or has the same gene location encoding the CD47 on the chromosome. Specifically, CD47 may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of SEQ ID NO: 149.

[0244] In one specific embodiment of the present invention, the second domain may include a heavy chain variable region and a light chain variable region of the anti-CD47 antibody Candidate14. Specifically, it may include a heavy chain variable region including HCDR1 comprising the amino acid sequence of SEQ ID NO: 809, HCDR2 comprising the amino acid sequence of SEQ ID NO: 811, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 813, and a light chain variable region including LCDR1 comprising the amino acid sequence of SEQ ID NO: 918, LCDR2 comprising the amino acid sequence of SEQ ID NO: 920, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 922. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 188, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 189 (C3). Alternatively, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 190, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 191 (C4).

[0245] In one specific example, the second domain may include a heavy chain variable region and a light chain variable region of the anti-CD47 antibody Magrolimab. Specifically, it may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 815, HCDR2 comprising the amino acid sequence of SEQ ID NO: 817, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 819, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 926, LCDR2 comprising the amino acid sequence of SEQ ID NO: 920, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 929. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 192, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 193 (C5). Alternatively, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 194, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 195 (C6).

[0246] In one specific example, the second domain may include a heavy chain variable region and a light chain variable region of the anti-CD47 antibody CC-90002. Specifically, it may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 821, HCDR2 comprising the amino acid sequence of SEQ ID NO: 823, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 825, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 932, LCDR2 comprising the amino acid sequence of SEQ ID NO: 934, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 936. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 196, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 197 (C7). Alternatively, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 198, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 199 (C8).

[0247] - EGFR

[0248] More specifically, the second domain may be an antibody or a fragment thereof that binds to epidermal growth factor receptor (EGFR).

[0249] As used herein, the term "EGFR" refers to a protein belonging to the ErbB (HER / EGFR / ERBB) family, also known as HER1 or ErbB-1, as described above. EGFR regulates cell growth, division, survival, and apoptosis. Overexpression of EGFR has been reported in tumor tissues of various cancers, and tumor tissues with increased EGFR are known to exhibit increased invasion, metastasis, and anticancer drug resistance.

[0250] In the present invention, EGFR may be included without limitation as long as it is derived from mammals, including primates such as humans and monkeys, and rodents such as rats and mice. In addition, the EGFR protein may include both a native or mutant EGFR protein, but is not limited thereto. The native EGFR protein generally refers to a polypeptide containing the amino acid sequence of the native EGFR protein. The amino acid sequence and polynucleotide sequence for the EGFR protein can be obtained from known databases such as the GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably, it may include the amino acid sequence of SEQ ID NO: 150.

[0251] Additionally, it may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted, as long as it has the same activity as the protein or has the same gene location encoding the EGFR on the chromosome. Specifically, the EGFR may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of SEQ ID NO: 150.

[0252] In one specific embodiment of the present invention, the second domain may include a heavy chain variable region and a light chain variable region of Cetuximab, an anti-EGFR antibody. Specifically, it may include a heavy chain variable region including HCDR1 comprising the amino acid sequence of SEQ ID NO: 829, HCDR2 comprising the amino acid sequence of SEQ ID NO: 832, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 834, and a light chain variable region including LCDR1 comprising the amino acid sequence of SEQ ID NO: 939, LCDR2 comprising the amino acid sequence of SEQ ID NO: 941, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 943. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 200, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 201 (C9). Alternatively, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 202, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 203 (C10).

[0253] In one specific example, the second domain may include a heavy chain variable region and a light chain variable region of the anti-EGFR antibody Panitumumab. Specifically, it may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 838, HCDR2 comprising the amino acid sequence of SEQ ID NO: 840, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 842, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 903, LCDR2 comprising the amino acid sequence of SEQ ID NO: 566, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 906. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 204, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 205 (C11). In this case, the heavy chain variable region may be located in the light chain, and the light chain variable region may be located in the heavy chain (C30). Alternatively, the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO: 206, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO: 207 (C12).

[0254] In one specific example, the second domain may include a heavy chain variable region and a light chain variable region of the anti-EGFR antibody Necitumumab. Specifically, it may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 844, HCDR2 comprising the amino acid sequence of SEQ ID NO: 846, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 848, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 580, LCDR2 comprising the amino acid sequence of SEQ ID NO: 566, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 937. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 208, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 209 (C13). Alternatively, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 210, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 211 (C14).

[0255] - CTLA4

[0256] More specifically, the second domain may be an antibody or fragment thereof that binds to cytotoxic T lymphocyte-associated antigen 4 (CTLA-4).

[0257] As used herein, the term "CTLA4" refers to an immune checkpoint protein, also known as CD152. CTLA4 shares homology with CD28 and competitively binds to B7-1 (CD80) or B7-2 (CD86) on the surface of antigen-presenting cells (APCs). When CD28 binds to CD80 or CD86, an immune response is activated, whereas when CTLA4 binds to CD80 or CD86, an immune response is suppressed. CTLA4 is known to suppress immune responses by binding to CD80 or CD86. In particular, CTLA4 is known to exhibit greater affinity and binding affinity for CD80 or CD86 compared to CD28.

[0258] In the present invention, CTLA4 may be included without limitation as long as it is derived from mammals, including humans, primates such as monkeys, and rodents such as rats and mice. In addition, the CTLA4 protein may include both a native or mutant CTLA4 protein, but is not limited thereto. The native CTLA4 protein generally refers to a polypeptide comprising the amino acid sequence of the native CTLA4 protein. The amino acid sequence and polynucleotide sequence for the CTLA4 protein can be obtained from known databases such as the GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably may include the amino acid sequence of SEQ ID NO: 151.

[0259] Additionally, it may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted, as long as it has the same activity as the protein or has the same gene location encoding the CTLA4 on the chromosome. Specifically, CTLA4 may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of SEQ ID NO: 151.

[0260] In one specific embodiment of the present invention, the second domain may include a heavy chain variable region and a light chain variable region of Ipilimumab, an anti-CTLA4 antibody. Specifically, it may include a heavy chain variable region including HCDR1 comprising an amino acid sequence of SEQ ID NO: 850, HCDR2 comprising an amino acid sequence of SEQ ID NO: 852, and HCDR3 comprising an amino acid sequence of SEQ ID NO: 854, and a light chain variable region including LCDR1 comprising an amino acid sequence of SEQ ID NO: 948, LCDR2 comprising an amino acid sequence of SEQ ID NO: 949, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 951. In this case, the heavy chain variable region may include an amino acid sequence of SEQ ID NO: 212, and the light chain variable region may include an amino acid sequence of SEQ ID NO: 213 (C15).

[0261] In one specific example, the second domain may include a heavy chain variable region and a light chain variable region of Tremelimumab, an anti-CTLA4 antibody. Specifically, it may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 855, HCDR2 comprising the amino acid sequence of SEQ ID NO: 857, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 858, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 952, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 954. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 214, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 215 (C16).

[0262] - B7H3

[0263] More specifically, the second domain may be an antibody or fragment thereof that binds to B7H3. The B7H3 is the same as described above.

[0264] In one specific embodiment of the present invention, the second domain may include a heavy chain variable region and a light chain variable region of an anti-B7H3 antibody (m8524). Specifically, it may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 757, HCDR2 comprising the amino acid sequence of SEQ ID NO: 759, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 761, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 580, LCDR2 comprising the amino acid sequence of SEQ ID NO: 566, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 799. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 139, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 140 (C17). Alternatively, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 216, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 217 (C18).

[0265] - ANXA1

[0266] More specifically, the second domain may be an antibody or fragment thereof that binds to ANXA1. The ANXA1 is the same as described above.

[0267] In the present invention, the second domain may include the heavy chain variable region and light chain variable region of A55 of UNIT1. The heavy chain variable region may include the amino acid sequence of SEQ ID NO: 109, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 110 (C19). A55 of UNIT1 is the same as described above.

[0268] - DLK1

[0269] More specifically, the second domain may be an antibody or fragment thereof that binds to DLK1 (Delta-like 1 homolog).

[0270] As used herein, the term "DLK1" refers to a transmembrane protein that is a member of the Notch / delta / serrate family.

[0271] DLK1 comprises an extracellular domain, a transmembrane domain, and an intracellular domain. The extracellular domain contains six EGF-like repeats. Furthermore, DLK1 is known to function independently, with the extracellular domain shed from the cell membrane by tumor necrosis factor alpha converting enzyme (TACE). Overexpression of DLK1 has been reported in brain and liver cancers.

[0272] In the present invention, DLK1 may be included without limitation as long as it is derived from mammals, including primates such as humans and monkeys, and rodents such as rats and mice. In addition, the DLK1 protein may include both a native or mutant DLK1 protein, but is not limited thereto. The native DLK1 protein generally refers to a polypeptide including the amino acid sequence of the native DLK1 protein. The amino acid sequence and polynucleotide sequence for the DLK1 protein can be obtained from known databases such as GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably may include the amino acid sequence of SEQ ID NO: 152.

[0273] Additionally, as long as it has the same activity as the protein or has the same gene location encoding the DLK1 on the chromosome, it may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted. Specifically, DLK1 may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of SEQ ID NO: 152.

[0274] In one specific embodiment of the present invention, the second domain may include a heavy chain variable region and a light chain variable region of an anti-DLK1 antibody (18A5). Specifically, it may include a heavy chain variable region including HCDR1 comprising an amino acid sequence of SEQ ID NO: 861, HCDR2 comprising an amino acid sequence of SEQ ID NO: 863, and HCDR3 comprising an amino acid sequence of SEQ ID NO: 865, and a light chain variable region including LCDR1 comprising an amino acid sequence of SEQ ID NO: 957, LCDR2 comprising an amino acid sequence of SEQ ID NO: 958, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 960. In this case, the heavy chain variable region may include an amino acid sequence of SEQ ID NO: 218, and the light chain variable region may include an amino acid sequence of SEQ ID NO: 219 (C20).

[0275] - CD3

[0276] More specifically, the second domain may be an antibody or fragment thereof that binds to CD3.

[0277] As used herein, the term "CD3" refers to a homodimeric or heterodimeric protein expressed on T cells in relation to the T cell receptor complex, which is an essential element for T cell activation. Functional CD3 is formed by dimeric association of two or more of four different chains, such as ε, ζ, δ, and γ, and CD3 dimeric configurations include γ / ε, δ / ε, and ζ / ζ. Antibodies to CD3 are known to bind to CD3 present on T cells and induce T cell activation.

[0278] In the present invention, CD3 may be included without limitation as long as it is derived from mammals, including primates such as humans and monkeys, and rodents such as rats and mice. In addition, the CD3 protein may include both native and mutant CD3 proteins, but is not limited thereto. The native CD3 protein generally refers to a polypeptide comprising the amino acid sequence of the native CD3 protein. The amino acid sequence and polynucleotide sequence for the CD3 protein can be obtained from known databases such as the GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably may include the amino acid sequence of SEQ ID NO: 153.

[0279] Additionally, it may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted, as long as it has the same activity as the protein or has the same gene location encoding the CD3 on the chromosome. Specifically, CD3 may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of SEQ ID NO: 153.

[0280] In one specific embodiment of the present invention, the second domain may include a heavy chain variable region and a light chain variable region of an anti-CD3 antibody (SP34#8). Specifically, it may include a heavy chain variable region including HCDR1 comprising an amino acid sequence of SEQ ID NO: 866, HCDR2 comprising an amino acid sequence of SEQ ID NO: 868, and HCDR3 comprising an amino acid sequence of SEQ ID NO: 870, and a light chain variable region including LCDR1 comprising an amino acid sequence of SEQ ID NO: 962, LCDR2 comprising an amino acid sequence of SEQ ID NO: 964, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 966. In this case, the heavy chain variable region may include an amino acid sequence of SEQ ID NO: 220, and the light chain variable region may include an amino acid sequence of SEQ ID NO: 221 (C21). Alternatively, the heavy chain variable region may include an amino acid sequence of SEQ ID NO: 232, and the light chain variable region may include an amino acid sequence of SEQ ID NO: 233 (C29).

[0281] - CLDN18.2

[0282] More specifically, the second domain may be an antibody or fragment thereof that binds to CLDN18.2 (Claudin 18.2).

[0283] As used herein, the term "CLDN18.2" refers to one of the isoforms of Claudin 18, a transmembrane protein that is a key component of epithelial cells and epithelial tight junctions that regulate the flow of substances between cells. CLDN18.2 is expressed in a restricted manner in the stomach, and is known to maintain the barrier function of the gastric mucosa and prevent hydrogen ion leakage from gastric acid. Overexpression has been reported in several types of tumors, including pancreatic cancer, esophageal cancer, ovarian adenocarcinoma, and lung cancer.

[0284] In the present invention, CLDN18.2 may be included without limitation as long as it is derived from mammals, including primates such as humans and monkeys, and rodents such as rats and mice. In addition, the CLDN18.2 protein may include both a native or mutant CLDN18.2 protein, but is not limited thereto. The native CLDN18.2 protein generally refers to a polypeptide comprising the amino acid sequence of the native CLDN18.2 protein. The amino acid sequence and polynucleotide sequence for the CLDN18.2 protein can be obtained from known databases such as the GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably may include the amino acid sequence of SEQ ID NO: 154.

[0285] Additionally, it may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted, as long as it has the same activity as the protein or has the same gene location encoding the CLDN18.2 on the chromosome. Specifically, CLDN18.2 may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of SEQ ID NO: 154.

[0286] In one specific embodiment of the present invention, the second domain may include a heavy chain variable region and a light chain variable region of the anti-CLDN18.2 antibody Zolbetuximab. Specifically, it may include a heavy chain variable region including HCDR1 comprising the amino acid sequence of SEQ ID NO: 872, HCDR2 comprising the amino acid sequence of SEQ ID NO: 874, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 876, and a light chain variable region including LCDR1 comprising the amino acid sequence of SEQ ID NO: 968, LCDR2 comprising the amino acid sequence of SEQ ID NO: 559, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 971. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 222, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 223 (C22).

[0287] In one specific example, the second domain may comprise a heavy chain variable region and a light chain variable region of the anti-CLDN18.2 antibody PR002726. Specifically, it may comprise a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 878, HCDR2 comprising the amino acid sequence of SEQ ID NO: 880, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 882, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 580, LCDR2 comprising the amino acid sequence of SEQ ID NO: 566, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 973. In this case, the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO: 224, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO: 225 (C23).

[0288] - TEM8

[0289] More specifically, the second domain may be an antibody or fragment thereof that binds to TEM8. The TEM8 is the same as described above.

[0290] In the present invention, the second domain may include the heavy chain variable region and light chain variable region of A70 of UNIT1. At this time, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 127, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 128 (C24). The A70 of UNIT1 is the same as described above.

[0291] - FAPα

[0292] More specifically, the second domain may be an antibody or fragment thereof that binds to FAPα. The FAPα is the same as described above.

[0293] In the present invention, the second domain may include the heavy chain variable region and light chain variable region of A75 of UNIT1. At this time, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 137, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 138 (C25). A75 of UNIT1 is the same as described above.

[0294] - EpCAM

[0295] More specifically, the second domain may be an antibody or fragment thereof that binds to an epithelial cell adhesion molecule (EpCAM).

[0296] The term "EpCAM" used in this specification refers to a type of cell adhesion molecule that exists in epithelial tissue and exhibits specific adhesiveness to its homologous receptor.

[0297] The above cell adhesion molecule (CAM) is a general term for molecules such as fibronectin, collagen, and laminin that make up the extracellular matrix, and receptors present on the cell surface that accept the molecules. Among these, cadherin and NCAM (neural cell adhesion molecule) show specific adhesiveness toward receptors of the same type, but most cell adhesion molecules except these are known to show adhesive activity toward adhesive molecules of different types.

[0298] In the present invention, EpCAM may be included without limitation as long as it is derived from mammals, including primates such as humans and monkeys, and rodents such as rats and mice. In addition, the EpCAM protein may include both a native or mutant EpCAM protein, but is not limited thereto. The native EpCAM protein generally refers to a polypeptide comprising the amino acid sequence of the native EpCAM protein. The amino acid sequence and polynucleotide sequence for the EpCAM protein can be obtained from known databases such as the GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably may include the amino acid sequence of SEQ ID NO: 155.

[0299] Additionally, it may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted, as long as it has the same activity as the protein or has the same gene location encoding the EpCAM on the chromosome. Specifically, EpCAM may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of SEQ ID NO: 155.

[0300] In one specific embodiment of the present invention, the second domain may include a heavy chain variable region and a light chain variable region of ING-1, which is an anti-EpCAM antibody. Specifically, it may include a heavy chain variable region including HCDR1 comprising an amino acid sequence of SEQ ID NO: 884, HCDR2 comprising an amino acid sequence of SEQ ID NO: 886, and HCDR3 comprising an amino acid sequence of SEQ ID NO: 888, and a light chain variable region including LCDR1 comprising an amino acid sequence of SEQ ID NO: 975, LCDR2 comprising an amino acid sequence of SEQ ID NO: 977, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 979. In this case, the heavy chain variable region may include an amino acid sequence of SEQ ID NO: 226, and the light chain variable region may include an amino acid sequence of SEQ ID NO: 227 (C26).

[0301] - PD-1

[0302] More specifically, the second domain may be an antibody or fragment thereof that binds to PD-1 (Programmed cell death-1).

[0303] As used herein, the term "PD-1," also known as CD279, refers to an immune checkpoint protein expressed on the surface of activated T cells (immune cells). Under normal circumstances, PD-1 mediates and maintains autoimmune tolerance of body tissues and plays a positive role in preventing the development of autoimmune diseases by preventing excessive activation of the immune system during inflammatory responses and damaging self-tissues. However, in pathological situations, PD-1 is known to be involved in the development and progression of tumor immunity and various autoimmune diseases. PD-1 is also expressed on B cells, natural killer cells, monocytes, and dendritic cells, and induces apoptosis of T cells by binding to PD-L1 or PD-L2, thereby weakening the cellular immune response.

[0304] In the present invention, PD-1 may be included without limitation as long as it is derived from mammals, including humans, primates such as monkeys, and rodents such as rats and mice. In addition, the PD-1 protein may include both a native or mutant PD-1 protein, but is not limited thereto. The native PD-1 protein generally refers to a polypeptide comprising the amino acid sequence of the native PD-1 protein. The amino acid sequence and polynucleotide sequence for the PD-1 protein can be obtained from known databases such as the GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably may include the amino acid sequence of SEQ ID NO: 156.

[0305] Additionally, it may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted, as long as it has the same activity as the protein or has the same gene location encoding the PD-1 on the chromosome. Specifically, PD-1 may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of SEQ ID NO: 156.

[0306] In one specific embodiment of the present invention, the second domain may include a heavy chain variable region and a light chain variable region of Nivolumab, an anti-PD-1 antibody. Specifically, it may include a heavy chain variable region including HCDR1 comprising an amino acid sequence of SEQ ID NO: 890, HCDR2 comprising an amino acid sequence of SEQ ID NO: 892, and HCDR3 comprising an amino acid sequence of SEQ ID NO: 894, and a light chain variable region including LCDR1 comprising an amino acid sequence of SEQ ID NO: 580, LCDR2 comprising an amino acid sequence of SEQ ID NO: 566, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 981. In this case, the heavy chain variable region may include an amino acid sequence of SEQ ID NO: 228, and the light chain variable region may include an amino acid sequence of SEQ ID NO: 229 (C27).

[0307] - PD-L1

[0308] More specifically, the second domain may be an antibody or fragment thereof that binds to PD-L1 (Programmed cell death ligand 1).

[0309] As used herein, the term "PD-L1" refers to a protein expressed on the surface of tumor cells or hematopoietic cells. It is also known as CD274 or B7H1. PD-L1 is a ligand for PD-1. When PD-L1 and PD-1 bind, they induce apoptosis of T cells, thereby weakening their immune activity.

[0310] In the present invention, PD-L1 may be included without limitation as long as it is derived from mammals, including humans, primates such as monkeys, and rodents such as rats and mice. In addition, the PD-L1 protein may include both a native or mutant PD-L1 protein, but is not limited thereto. The native PD-L1 protein generally refers to a polypeptide including the amino acid sequence of the native PD-L1 protein. The amino acid sequence and polynucleotide sequence for the PD-L1 protein can be obtained from known databases such as the GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably may include the amino acid sequence of SEQ ID NO: 157.

[0311] In addition, it may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted, as long as it has the same activity as the protein or the gene location encoding the PD-L1 on the chromosome is the same. Specifically, PD-L1 may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of SEQ ID NO: 157.

[0312] In one specific example, the second domain may include a heavy chain variable region and a light chain variable region of the anti-PD-L1 antibody Atezolizumab. Specifically, it may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 895, HCDR2 comprising the amino acid sequence of SEQ ID NO: 898, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 900, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 982, LCDR2 comprising the amino acid sequence of SEQ ID NO: 680, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 984. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 230, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 231 (C28). Alternatively, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 237, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 236. At this time, the second domain heavy chain variable region may be located in the light chain and the light chain variable region may be located in the heavy chain, or a fragment thereof (C31).

[0313] - Amyloid beta protofibrils

[0314] More specifically, the second domain may be an antibody or fragment thereof that binds to amyloid beta protofibrils (Aβ protofibrils).

[0315] As used herein, the term "Amyloid beta (Aβ) protofibrils" refers to an intermediate form of Aβ aggregates that are generated in the cascade of aggregation types from Aβ monomers to Aβ plaques. Although amyloid plaques composed of fibrillar deposits of Aβ are a hallmark of Alzheimer's disease, soluble Aβ, particularly oligomers and protofibrils, measured in the brain tissue and cerebrospinal fluid of postmortem Alzheimer's patients are more associated with disease progression and are reported to be detrimental to synapses and neurons.

[0316] In the present invention, Aβ protofibrils may be included without limitation as long as they are derived from mammals, including primates such as humans and monkeys, and rodents such as rats and mice.

[0317] The amino acid sequence and polynucleotide sequence for the above Aβ protofibril can be obtained from a known database such as the GenBank of the National Center for Biotechnology Information (NCBI) in the United States, and preferably includes the amino acid sequence of SEQ ID NO: 158.

[0318] Additionally, it may be composed of a sequence in which one or more amino acids of the protein are added, deleted, or substituted, as long as it has the same activity as the protein or has the same gene location encoding the amyloid beta on the chromosome. Specifically, the amyloid beta may comprise or consist of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity, with the amino acid sequence of SEQ ID NO: 158.

[0319] In one specific embodiment of the present invention, the second domain may include a heavy chain variable region and a light chain variable region of Lecanemab, an anti-Aβ protofibril antibody. Specifically, it may include a heavy chain variable region including HCDR1 comprising the amino acid sequence of SEQ ID NO: 908, HCDR2 comprising the amino acid sequence of SEQ ID NO: 910, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 912, and a light chain variable region including LCDR1 comprising the amino acid sequence of SEQ ID NO: 986, LCDR2 comprising the amino acid sequence of SEQ ID NO: 920, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 989. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 239, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 240 (C33).

[0320] In the present invention, the immune modulator, antibody or fragment thereof, or antigen binding site forming the second domain is named “UNIT2.”

[0321] Linker

[0322] In the present invention, the first domain and the second domain of the fusion protein may be connected by a linker. In this case, the linker may be a first linker. Preferably, it may be a type 1 cleavable linker.

[0323] The ReleaDome of the present invention can exist in an inactive state at a non-disease site by having the first domain and the second domain connected by the first-type cleavable linker, whereby the binding affinity of the second domain is shielded by the first domain. In addition, when the ReleaDome is located at a disease site to which the first domain specifically binds, the cleavable linker is cleaved by a protease specifically expressed at the disease site, thereby dissociating the first domain and allowing the second domain to bind to the target, thereby exhibiting a pharmacological effect.

[0324] First linker

[0325] The first linker may be a peptide linker. Furthermore, the first linker may be a cleavable linker or a non-cleavable linker. Hereinafter, the cleavable linker of the first linker is referred to as a "type 1 cleavable linker," and the non-cleavable linker is referred to as a "type 1 non-cleavable linker." Furthermore, the first linker is often used interchangeably with "second component" or "B" in the present specification.

[0326] As used herein, the term "peptide linker" refers to a peptide composed of one or more amino acids. Typically, the peptide may be composed of 1 to 100 consecutive amino acids, 1 to 50 consecutive amino acids, 3 to 30 consecutive amino acids, or 5 to 15 amino acids. Peptide linkers are known in the art or described herein.

[0327] - Type 1 cleavable linker

[0328] As used herein, the term "type 1 cleavable linker" refers to a polypeptide linker comprising a cleavable peptide. The cleavable peptide is a polypeptide comprising a protease cleavage site, and can be specifically cleaved by a protease. As used herein, the "cleavable peptide" may be described interchangeably with the "substrate peptide." The protease is an enzyme that hydrolyzes a peptide bond between two specific amino acid residues within a target substrate protein. Accordingly, the "cleavage site" refers to an amino acid sequence within a target substrate protein that is recognized and cleaved by a protease, and the cleavable peptide may comprise the amino acid sequence of such a protease cleavage site (substrate sequence). The cleavable peptide is generally cleaved by an enzyme, typically a peptidase, protease, nuclease, lipase, or the like. Additionally, cleavable peptides can be cleaved by changes in environmental signals, such as temperature, pH, salt concentration, etc.

[0329] In the present invention, the cleavable peptide may include an amino acid sequence of a cleavage site that is cleaved by a protein decomposition enzyme that is specifically overexpressed or overactivated in a diseased site compared to a non-diseased site.

[0330] In one specific example, the cleavable peptide may comprise an amino acid sequence of a tumor-associated protease cleavage site. The tumor-associated protease may be a protease overexpressed in a tumor cell or tumor environment.

[0331] In one specific example, the cleavable peptide may comprise an amino acid sequence of a cleavage site of a protease associated with a degenerative brain disease (e.g., Alzheimer's disease). Here, the degenerative brain disease-associated protease may be a protease that is overexpressed in an environment where a degenerative brain disease has developed.

[0332] In the present invention, the masking effect of the first domain (first component) that shields the second domain (third component) may vary depending on the length of the cleavable peptide and the amino acid composition and arrangement.

[0333] The above cleavable peptides may exhibit different cleavage efficiencies and may not be cleaved or may exhibit very weak cleavage efficiencies at non-diseased sites.

[0334] Specifically, the protein-degrading enzymes specifically expressed in the disease site include human Matrix metalloprotease (MMP)-1, MMP-2, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-9, MMP-10, MMP-11, MMP-12, MMP-13, MMP-14, MMP-15, MMP-16, MMP-17, MMP-18, MMP-19, MMP-20, MMP-21, MMP-22, MMP-23, MMP-24, MMP-25, MMP-26, MMP-27, MMP-28, human Cathepsin A, Cathepsin B, Cathepsin C, Cathepsin D, Cathepsin E, Cathepsin F, Cathepsin G, Cathepsin H, Cathepsin K, Cathepsin L1, Cathepsin V, Cathepsin O, Cathepsin S, Cathepsin W, Cathepsin Z, human Eenterokinase (EK), human Thrombin (Thr), human tPA (, tissue plasminogen activator), human Granzyme BGrB), human uPA (urokinase-type plasminogen activator), human ADAMTs-5 (A disintegrin-like and metalloproteinase with thrombospondin type1 motif 5, A5), Kallikreins, human K1, human K10, human Plasmin, Collagenase, Type IV collagenase, Sromelysin, Factor Mirl CP, Papain, HIV-1 protease,It may be any one selected from the group consisting of HSV protease, CMV protease, Chymosin, Renin, Pepsin, Matriptase, Legumain, Plasmepsin, Nepenthesin, Metalloexopeptidases, Metalloendopeptidases, Prostate specific antigen (PSA / hK3), Interleukin-1β converting enzyme, Thrombin, FAPα, Dipeptidyl peptidase, Type II transmembrane serine proteases (TTSP), Neutrophil serine protease, Proteinase 3, Neutrophil serine protease4, Mast cell chymase, Mast cell tryptases, and combinations thereof, but is not limited thereto.

[0335] The above cleavable peptide may be composed of 2 to 20 consecutive amino acids. Specifically, the above cleavable peptide may be composed of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 consecutive amino acids.

[0336] Additionally, in the present invention, the cleavable peptide may include a variant. In this case, the variant (or PRS variant) refers to a cleavable peptide comprising a substrate sequence for one or more proteolytic enzymes. In this case, the composition and arrangement of the substrate sequences of each included proteolytic enzyme may be combined differently. Accordingly, the proteolytic enzyme that recognizes the cleavable peptide variant may be one or more types, and the cleavage efficiency of the substrate peptide may vary depending on the substrate sequence and the proteolytic enzyme.

[0337] More specifically, the cleavable peptide may be cleaved by any one protease selected from the group consisting of MMP-2, MMP-9, uPA, MT-SP1, MMP-3, MMP-7, and combinations thereof. In one specific embodiment of the present invention, the cleavable peptide may comprise any one amino acid selected from the group consisting of SEQ ID NO: 159 to SEQ ID NO: 176.

[0338] In one embodiment, the cleavable peptide can be cleaved by MMP-2. Specifically, the cleavable peptide can comprise an amino acid sequence of SEQ ID NO: 159 to SEQ ID NO: 161, SEQ ID NO: 163 to SEQ ID NO: 170, SEQ ID NO: 173, or SEQ ID NO: 174. In one embodiment, the cleavable peptide can be cleaved by MMP-9. Specifically, the cleavable peptide can comprise an amino acid sequence of SEQ ID NO: 163 to SEQ ID NO: 166, SEQ ID NO: 169, or SEQ ID NO: 170. In one embodiment, the cleavable peptide can be cleaved by uPA. In one embodiment, the cleavable peptide can be cleaved by MT-SP1. In one embodiment, the cleavable peptide can be cleaved by MMP-3. Specifically, the cleavable peptide may comprise an amino acid sequence of SEQ ID NO: 163 to SEQ ID NO: 166. In one specific example, the cleavable peptide may be cleaved by MMP-7. Specifically, the cleavable peptide may comprise an amino acid sequence of SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 163 to SEQ ID NO: 166, or SEQ ID NO: 174.

[0339] In one embodiment, the cleavable peptide can be cleaved by MMP-2 and MMP-9. Specifically, the cleavable peptide can comprise an amino acid sequence of SEQ ID NO: 159 to SEQ ID NO: 161, SEQ ID NO: 163 to SEQ ID NO: 170, SEQ ID NO: 173, or SEQ ID NO: 174. In one embodiment, the cleavable peptide can be cleaved by MMP-2 and uPA. Specifically, the cleavable peptide can comprise an amino acid sequence of SEQ ID NO: 159 to SEQ ID NO: 161, SEQ ID NO: 163 to SEQ ID NO: 170, SEQ ID NO: 173, or SEQ ID NO: 174. In one embodiment, the cleavable peptide can be cleaved by MMP-2 and MT-SP1. Specifically, the cleavable peptide may include an amino acid sequence of SEQ ID NO: 159 to SEQ ID NO: 161, SEQ ID NO: 163 to SEQ ID NO: 170, SEQ ID NO: 173, or SEQ ID NO: 174. In one embodiment, the cleavable peptide may be cleaved by MMP-9 and uPA. Specifically, the cleavable peptide may include an amino acid sequence of SEQ ID NO: 163 to SEQ ID NO: 166, SEQ ID NO: 169, or SEQ ID NO: 170. In one embodiment, the cleavable peptide may be cleaved by MMP-9 and MT-SP1. Specifically, the cleavable peptide may include an amino acid sequence of SEQ ID NO: 163 to SEQ ID NO: 166, SEQ ID NO: 169, or SEQ ID NO: 170. In one embodiment, the cleavable peptide may be cleaved by uPA and MT-SP1. Specifically, the cleavable peptide may comprise an amino acid sequence of SEQ ID NO: 161 to SEQ ID NO: 168. In one specific example, the cleavable peptide may be cleaved by MMP-2, MMP-9, and uPA. Specifically, the cleavable peptide may comprise an amino acid sequence of SEQ ID NO: 159 to SEQ ID NO: 161, SEQ ID NO: 163 to SEQ ID NO: 170, SEQ ID NO: 173, or SEQ ID NO: 174.In one embodiment, the cleavable peptide can be cleaved by MMP-2, uPA, and MT-SP1. Specifically, the cleavable peptide can comprise an amino acid sequence of SEQ ID NO: 159 to SEQ ID NO: 170, SEQ ID NO: 173, or SEQ ID NO: 174. In one embodiment, the cleavable peptide can be cleaved by MMP-9, uPA, and MT-SP1. Specifically, the cleavable peptide can comprise an amino acid sequence of SEQ ID NO: 161 to SEQ ID NO: 170. In one embodiment, the cleavable peptide can be cleaved by MMP-2, MMP-9, uPA, and MT-SP1. Specifically, the cleavable peptide can comprise an amino acid sequence of SEQ ID NO: 159 to SEQ ID NO: 170, SEQ ID NO: 173, or SEQ ID NO: 174. In one specific example, the cleavable peptide can be cleaved by MMP-3 and MMP-7. Specifically, the cleavable peptide can comprise an amino acid sequence of SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 163 to SEQ ID NO: 166, or SEQ ID NO: 174.

[0340] In one embodiment, the cleavable peptide can be cleaved by MMP-2, MMP-9, MMP-3, and MMP-7. Specifically, the cleavable peptide can comprise an amino acid sequence of SEQ ID NO: 159 to SEQ ID NO: 161, SEQ ID NO: 163 to SEQ ID NO: 170, SEQ ID NO: 173, or SEQ ID NO: 174. In one embodiment, the cleavable peptide can be cleaved by MMP-2, uPA, MMP-3, and MMP-7. Specifically, the cleavable peptide can comprise an amino acid sequence of SEQ ID NO: 159 to SEQ ID NO: 161, SEQ ID NO: 163 to SEQ ID NO: 170, SEQ ID NO: 173, or SEQ ID NO: 174. In one embodiment, the cleavable peptide can be cleaved by MMP-2, MT-SP1, MMP-3, and MMP-7. Specifically, the cleavable peptide may include an amino acid sequence of SEQ ID NO: 159 to SEQ ID NO: 161, SEQ ID NO: 163 to SEQ ID NO: 170, SEQ ID NO: 173, or SEQ ID NO: 174. In one embodiment, the cleavable peptide may be cleaved by MMP-9, uPA, MMP-3, and MMP-7. Specifically, the cleavable peptide may include an amino acid sequence of SEQ ID NO: 160 to SEQ ID NO: 170, or SEQ ID NO: 174. In one embodiment, the cleavable peptide may be cleaved by MMP-9, MT-SP1, MMP-3, and MMP-7. Specifically, the cleavable peptide may include an amino acid sequence of SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 163 to SEQ ID NO: 166, SEQ ID NO: 169, SEQ ID NO: 170, or SEQ ID NO: 174. In one specific example, the cleavable peptide can be cleaved by uPA, MT-SP1, MMP-3, and MMP-7. Specifically, the cleavable peptide can comprise an amino acid sequence of SEQ ID NO: 160 to SEQ ID NO: 168 or SEQ ID NO: 174.

[0341] In one embodiment, the cleavable peptide can be cleaved by MMP-2, MMP-9, uPA, MMP-3, and MMP-7. Specifically, the cleavable peptide can comprise an amino acid sequence of SEQ ID NO: 159 to SEQ ID NO: 161, SEQ ID NO: 163 to SEQ ID NO: 170, SEQ ID NO: 173, or SEQ ID NO: 174. In one embodiment, the cleavable peptide can be cleaved by MMP-2, uPA, MT-SP1, MMP-3, and MMP-7. Specifically, the cleavable peptide can comprise an amino acid sequence of SEQ ID NO: 159 to SEQ ID NO: 170, SEQ ID NO: 173, or SEQ ID NO: 174. In one embodiment, the cleavable peptide can be cleaved by MMP-9, uPA, MT-SP1, MMP-3, and MMP-7. Specifically, the cleavable peptide may comprise an amino acid sequence of SEQ ID NO: 160 to SEQ ID NO: 170 or SEQ ID NO: 174.

[0342] In one specific example, the cleavable peptide can be cleaved by MMP-2, MMP-9, uPA, MT-SP1, MMP-3, and MMP-7. Specifically, the cleavable peptide can comprise an amino acid sequence of SEQ ID NO: 159 to SEQ ID NO: 170, SEQ ID NO: 173, or SEQ ID NO: 174.

[0343] Table 5 below shows exemplary sequences of cleavable peptides that may be included in the cleavable linkers provided herein. The amino acid sequences are listed sequentially from the N-terminus to the C-terminus. Fusion proteins according to exemplary embodiments of the present invention may include these cleavable peptides.

[0344] Hereinafter, in the present invention, the type 1 cleavable linker including the cleavable peptide is described interchangeably with 'PRS (Protein recognition site)' or 'type 1 cleavable linker'.

[0345]

[0346]

[0347] In the present invention, the first type cleavable linker may include a space domain at the N-terminus and / or C-terminus of the cleavable peptide. The space domain is located at the N-terminus and / or C-terminus of the cleavable peptide, and may serve to position the peptide without a protease cleavage site so that the protease can more easily access the cleavage site of the cleavable peptide. The length of the space domain may be optimized for accessibility for enzymatic cleavage. The space domain may each include about 1 to 100 amino acids. Specifically, it may consist of 1 to 100, 1 to 80, 1 to 60, 1 to 50, or 1 to 30 amino acids.

[0348] - Type 1 non-cleavable linker

[0349] As used herein, the term "type 1 non-cleavable linker" refers to a peptide linker that does not contain a cleavage site that is cleaved by a protease. Such non-cleavable linkers are known in the art or are described herein. The type 1 non-cleavable linker may be comprised of 1 to 50 consecutive amino acids, or 3 to 30 consecutive amino acids, or 5 to 20 amino acids. The non-cleavable linker may be (G4S)n, and may further comprise (G)n. The non-cleavable linker may be (G)n or (GS3)n. In this case, n may each independently be 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In one embodiment of the present invention, the non-cleavable linker may comprise an amino acid sequence of SEQ ID NO: 261 to SEQ ID NO: 266. Table 6 below illustrates exemplary sequences that may be included in the non-cleavable linkers provided herein. The amino acid sequences below are listed sequentially from the N-terminus to the C-terminus. Fusion proteins according to exemplary embodiments of the present invention may include, but are not limited to, these sequences.

[0350] Hereinafter, in the present invention, the first type non-cleavable linker is described interchangeably with the “fifth component” or “E”.

[0351] 5th component Part Number Variant Amino acid sequence G4S linker E1 (G4S) 1 GGGGS (SEQ ID NO: 261) E2 (G4S) 2 GGGGSGGGGS (SEQ ID NO: 262) E3 (G4S) 3 GGGGSGGGGSGGGGS (SEQ ID NO: 263) E4 PCPAPELLGGP (G4S) 1 PCPAPELLGGPGGGGS (SEQ ID NO: 264) linker E5-PCPAPELLGGP (SEQ ID NO: 265) (G)n linker E6 (G) 7 GGGGGGG (SEQ ID NO: 266)

[0352] Second linker

[0353] In this specification, the second linker refers to a linker that connects the fusion protein and the payload of the present invention. The second linker may be a cleavable linker or a non-cleavable linker. The non-cleavable linker refers to a linker in which the fusion protein-drug conjugate is internalized into a target cell, and then the drug is released in a form bound to the linker through catabolism in the cytoplasm or lysosome. On the other hand, the cleavable linker refers to a linker in which the fusion protein-drug conjugate is internalized into a target cell, and then cleaved by a special environment or enzyme of a lysosome or endosome, thereby releasing the drug. Hereinafter, the cleavable linker may be referred to as a "type 2 cleavable linker," and the non-cleavable linker may be referred to as a "type 2 non-cleavable linker."

[0354] The above type 2 cleavable linkers can be classified according to the principle by which cleavage occurs.

[0355] The above-mentioned second type cleavable linker may be chemically cleavable or enzymatically cleavable.

[0356] In one specific example, the chemically cleavable linker may be an acid-labile linker or a reducible linker.

[0357] As used herein, the term "acid-labile linker" refers to a linker that is stable in a neutral pH, such as blood, but undergoes cleavage by acid hydrolysis in an acidic environment (pH 5 to pH 6), such as a cancer cell microenvironment, lysosome, or endosome. The acid-labile linker includes a hydrazone linker or an ester linker.

[0358] As used herein, the term "reducible linker" refers to a linker that undergoes cleavage upon reduction by a reducing agent. In one specific example, the reducible linker may be a disulfide linker.

[0359] As used herein, the term "enzyme-cleavable linker" refers to a linker that is cleaved by a reducing agent within a cell. The enzyme-cleavable linker may be a peptide-based linker or a specific enzyme-based linker.

[0360] As used herein, the term "peptide-based linker" refers to a linker that is cleaved by an enzyme that is relatively abundant within a cell, and that includes a specific peptide bond cleavage site. The peptide-based linker may be a linker that includes one or more binding sites selected from the group consisting of valine-citrulline, valine-alanine, and phenylalanine-glycine. In one specific example, the peptide-based linker may be, but is not limited to, a valine-citrulline linker, a valine-alanine linker, an alanine-alanine-alanine, or a phenylalanine-glycine linker.

[0361] As used herein, the term "enzyme-specific linker" refers to a linker designed to be cleaved by a specific enzyme. The enzyme-specific linker may be cleaved by one or more enzymes selected from the group consisting of β-galactoside, β-glucuronide, and phosphodiester. In one embodiment, the enzyme-specific linker may be one or more enzymes selected from the group consisting of β-galactoside linkers, β-glucuronide linkers, phosphodiester linkers, and combinations thereof.

[0362] In one specific example, the second type non-cleavable linker may be, but is not limited to, a Maleimidocaproyl (MC) linker or a Succimidyl 4-(N-aleimidomethyl)cyclohexane-1-carboxylate (SMCC) linker.

[0363] In one specific example, the cleavable linker may be any one selected from the group consisting of a hydrazone linker, an ester linker, a disulfide linker, a valine-citrulline linker, a valine-alanine linker, an alanine-alanin-alanin linker, a phenylalanine-glycine linker, a β-galactoside linker, a β-glucuronide linker, a phosphodiester linker, and combinations thereof.

[0364] In one embodiment, the linker may include a structural modification. In one embodiment, the linker may have polyethylene glycol (PGE) substituted in a portion of the structure, or may include a self-immolative chemical structure (SIG, Self-immolative group).

[0365] Third linker

[0366] In the present invention, the third linker may be a peptide linker composed of 5 to 80 consecutive amino acids, 7 to 70 consecutive amino acids, 10 to 60 consecutive amino acids, or 12 to 50 amino acids. The third linker may include at least one cysteine. Specifically, it may include one, two, or three cysteines. Additionally, the third linker may be derived from the hinge of an immunoglobulin.

[0367] The third linker may comprise a hinge of an immunoglobulin or a variant thereof, wherein the hinge may be modified to delete a disulfide bond or introduce an additional disulfide bond.

[0368] In one specific embodiment of the present invention, the third linker may comprise any one amino acid sequence selected from the group consisting of SEQ ID NO: 270 to SEQ ID NO: 275. Table 7 below lists exemplary IgG subtypes and fragments thereof having core hinge sequences that may be included in the third linker provided in the present invention. The amino acid sequences are listed sequentially from the N-terminus to the C-terminus.

[0369] Hereinafter, in the present invention, the third linker is described interchangeably with “hinge” or “core hinge.”

[0370] Third linker Part Number IgG subtype Amino acid sequence Core hinge (hinge / hinge variant) DHH1 IgG1 EPKSCDKTHTCPPCP (SEQ ID NO: 270) DHH2 IgG2 ERKCCVECPPCP (SEQ ID NO: 271) DHH3 IgG3 ELKTPLDTTHTCPRCP (EPKSCDTPPPCPRCP) 3 (SEQ ID NO: 272) DHH4 IgG4 ESKYGPPCPSCP (SEQ ID NO: 273) DHH5 IgG1-RBDKTHTCPPCP (SEQ ID NO: 274) DHH6 IgG4-Variant ESKYGPPCPPCP (SEQ ID NO: 275)

[0371] Structure of fusion proteins

[0372] Specifically, the fusion protein of the present invention may include the following structural formulas (I) and (II).

[0373] N'-T1'-((L1)a-CR1)b-(L2')-I1'-((L3)c-CR3)dC' (I);

[0374] N'-T1''-((L1)a-CR2)b-(L2'')-I1''-((L3)c-CR4)dC' (II)

[0375] At this time, in the structural formulas (I) and (II),

[0376] The above N' is the N-terminus of the fusion protein,

[0377] The above C' is the C-terminus of the fusion protein,

[0378] The above T1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the first target,

[0379] The above T1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the first target,

[0380] The above T1' and T1'' combine to form a binding site (T1) that specifically binds to the first target,

[0381] The above I1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target,

[0382] The above I1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target,

[0383] The above I1' and I1'' combine to form a binding site (I1) that specifically binds to the second target,

[0384] The above CR1, CR2, CR3 and CR4 are each independently CH1 or CL,

[0385] At this time, CR1 and CR2 are not equal, CR3 and CR4 are not equal,

[0386] The above CH1 is a heavy chain constant region,

[0387] The above CL is a light chain constant region,

[0388] The above L1, L2', L2'' and L3 are each independently a peptide linker, wherein L2' and L2'' are truncated or uncleaved,

[0389] The above a, b, c and d are each independently 0 or 1.

[0390] The first target, second target, heavy chain variable region, light chain variable region, heavy chain constant region, and light chain constant region are the same as described above.

[0391] The above L1 and L3 are each independently a type 1 non-cleavable linker, and L2' and L2'' can be a type 1 non-cleavable linker or a type 1 cleavable linker. The first linker, the type 1 cleavable linker, and the type 1 non-cleavable linker are the same as described above.

[0392] Specifically, the first structure of the structural formula (I) comprises a heavy chain variable region (VH1) or a light chain variable region (VL) of an antibody or a fragment thereof that specifically binds to a first target and a heavy chain variable region (VH2) or a light chain variable region (VL2) that specifically binds to a second target; the second structure of the structural formula (II) comprises a light chain variable region (VL1) or a heavy chain variable region (VH1) that specifically binds to the first target and a light chain variable region (VL2) or a heavy chain variable region (VH2) that specifically binds to the second target, and the first structure and the second structure can be combined to form a binding site that specifically binds to the first target and the second target.

[0393] At this time, the binding of the first structure and the second structure may be formed through i) a disulfide bond by cysteine ​​(Cys) present in the heavy chain constant region (CH1) and the light chain constant region (CL) of CR3 and CR4, ii) a disulfide bond by cysteine ​​(Cys) present in the heavy chain variable region (VH2) and the light chain variable region (VL2) within I1' and I1'', or iii) a disulfide bond by cysteine ​​(Cys) present in the heavy chain constant region (CH1) and the light chain constant region (CL) of CR3 and CR4 and a disulfide bond by cysteine ​​(Cys) present in the heavy chain variable region (VH2) and the light chain variable region (VL2) within I1' and I1''.

[0394] Specifically, the bond between the first structure and the second structure can be formed by a disulfide bond between the 103rd cysteine ​​of CH1 and the 107th cysteine ​​of CL based on the Kabat numbering. In addition, the first structure and the second structure can further include a cysteine ​​(Cys) through an amino acid substitution. An example of such a mutant may include a mutation in the variable region of the second domain. Specifically, a disulfide bond can be formed by substituting cysteine ​​at the 44th position of VH2 and the 100th position of VL2 based on the Kabat numbering.

[0395] Specifically, the fusion protein may include the following structural formulas (IV) and (V).

[0396] N'-T2-(L7)-I2'-((L8)f-CR1)gC' (IV);

[0397] N'-I2''-((L8)f-CR2)hC' (V)

[0398] At this time, in the structural formulas (IV) and (V),

[0399] The above N' is the N-terminus of the fusion protein,

[0400] The above C' is the C-terminus of the fusion protein,

[0401] The above T2 is a peptide or antibody fragment that specifically binds to the first target,

[0402] The above I2' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target,

[0403] The above I2'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target,

[0404] The above I2' and I2'' combine to form a binding site (I2) that specifically binds to the second target,

[0405] The above CR1 and CR2 are each independently CH1 or CL,

[0406] At this time, CR1 and CR2 are not equal,

[0407] The above CH1 is a heavy chain constant region,

[0408] The above CL is a light chain constant region,

[0409] The above L7 and L8 are each independently a peptide linker, wherein L7 is a cleavable or non-cleavable linker,

[0410] The above f, g and h are each independently 0 or 1, and wherein g and h are the same.

[0411] The first target, second target, heavy chain variable region, light chain variable region, heavy chain constant region, and light chain constant region are the same as described above.

[0412] The above L7 may be a first linker, and L8 may be a first-type non-cleavable linker. The first linker, the first linker, and the first-type non-cleavable linker are the same as described above.

[0413] Specifically, the fourth structure of the structural formula (IV) comprises a heavy chain variable region (VH1) of a peptide or antibody fragment that specifically binds to a first target and a heavy chain variable region (VH2) or a light chain variable region (VL2) of an antibody that specifically binds to a second target; and the fifth structure of the structural formula (V) comprises a light chain variable region (VL2) or a heavy chain variable region (VH2) of an antibody that specifically binds to a second target, wherein the fourth structure and the fifth structure can be combined to form a binding site that specifically binds to the first target and the second target.

[0414] At this time, the binding of the fourth structure and the fifth structure can be formed through i) a disulfide bond by cysteine ​​(Cys) present in the heavy chain constant region (CH1) and the light chain constant region (CL), ii) a disulfide bond by cysteine ​​(Cys) present in the heavy chain variable region (VH2) and the light chain variable region (VL2) within I2' and I2'', or iii) a disulfide bond by cysteine ​​(Cys) present in the heavy chain constant region (CH1) and the light chain constant region (CL) and a disulfide bond by cysteine ​​(Cys) present in the heavy chain variable region (VH2) and the light chain variable region (VL2) within I2' and I2''.

[0415] Specifically, the bonding of the fourth and fifth structures can be formed by a disulfide bond between the 103rd cysteine ​​of CH1 and the 107th cysteine ​​of CL based on Kabat numbering. In addition, the fourth and fifth structures can further include cysteine ​​(Cys) through amino acid substitution. An example of such a mutant may include a mutation in the variable region of the second domain. Specifically, a disulfide bond can be formed by substituting cysteine ​​at the 44th position of VH2 and the 100th position of VL2 based on Kabat numbering.

[0416] Lilydom containing the seventh component

[0417] In the present invention, the fusion protein may additionally include a payload. In this case, the fusion protein and the payload may be linked via a linker. Specifically, the payload may be linked to the fusion protein via a linker at the C-terminus of the light chain constant region of the second domain of the fusion protein. The payload may be an anticancer agent, and the linker may be a second linker.

[0418] In this specification, the second linker and payload are referred to as the "seventh component" and are described interchangeably with "UNIT3." In addition, a fusion protein comprising the seventh component is described interchangeably with the term "fusion protein-drug conjugate."

[0419] Additionally, the fusion protein may further comprise a peptide motif to enhance binding with the seventh component. In this case, the fusion protein and the peptide motif may be linked via a first non-cleavable linker, and the peptide motif may bind to the seventh component via an enzymatic reaction.

[0420] In one specific example, the second domain of the fusion protein may comprise a type 1 non-cleavable linker and a peptide motif at the C-terminus of the light chain constant region (CL), wherein the peptide motif can bind to the seventh component through an enzymatic reaction.

[0421] - Peptide motif

[0422] The term "peptide motif" as used herein refers to an amino acid motif that can be recognized by an enzyme. In this case, the enzyme may be, but is not limited to, sortase, formylglycine generating enzyme (FGE), transglutaminase (TG), or isoprenoid transferase.

[0423] The above "sortase" is an enzyme derived from Gram-positive bacteria that recognizes the amino acid sequence 'LPXTG' and catalyzes the replacement of the terminal glycine (Gly) with a conjugation partner comprising an N-terminal glycine residue. Therefore, a fusion protein comprising the above amino acid motif at the C-terminus can be conjugated with a glycine-functionalized payload to form various fusion protein-drug conjugates. The amino acid motif can be LPXTG, LPXAG, LPXSG, LAXTG, LPXTG, LPXTA or NPQTG (wherein X is any amino acid, and preferably X is E or Q).

[0424] The above "FGE" recognizes the amino acid sequence of the amino acid motif "LCXPXR" (wherein X is serine (Ser), threonine (Thr), alanine (Ala), or glycine (Gly)) and oxidizes and converts cysteine ​​(Cys) in the sequence into formyl-glycine having an aldehyde group. Therefore, a fusion protein including the above amino acid motif can form a Hydrazino-iso-Pictet-Spengler (HIPS) ligation to selectively form a fusion protein-drug conjugate at various positions.

[0425] The above "transglutaminase" generally catalyzes the formation of an amide bond between a glutamine (Gln) side chain and a primary amine in a finite small molecule. Among the above enzymes, the transglutaminase from Streptoverticillium mobaraense recognizes glutamine (Gln) within the "LLQG" amino acid sequence. Therefore, a fusion protein comprising the above amino acid motif can selectively form a fusion protein-drug conjugate at various positions.

[0426] The above "isoprenoid transferase" refers to an enzyme that can recognize a specific amino acid motif at or near the C-terminus of a protein and can selectively alkylate the thiol position of cysteine ​​(Cys) residue(s) of the specific amino acid motif by adding isoprenoid unit(s) to the protein having the specific amino acid motif. Examples of the above isoprenoid transferase may include farnesyl transferase (FTase) and geranylgeranyl transferase (GGTase). Each of the above enzymes involves the transfer of a farnesyl or geranyl-geranyl residue to the C-terminal cysteine(s) of the target protein, respectively. At this time, the protein to which the C-terminal cysteine ​​of the target protein is transferred as described above is called a functionalized protein. GGTase can be classified into GGTase I and GGTase II. FTase and GGTase I can recognize the CAAX motif, and GGTase II can recognize the XXCC, XCXC, or CXX motif (wherein C is cysteine, A is an aliphatic amino acid, and X is an amino acid that determines the substrate specificity of the isoprenoid transferase). In addition, isoprenoid transferase can recognize not only substrates but also isosubstrates. An isosubstrate refers to a substrate analog that has a modification in the substrate. A fusion protein containing the above amino acid motif at the C-terminus can be conjugated to a payload to form various fusion protein-drug conjugates.

[0427] In one specific example, the peptide motif may comprise a CAAX motif, and more specifically, may comprise the amino acid sequence CVIM (SEQ ID NO: 267). In this case, the first type non-cleavable linker may be GGGGGGG (SEQ ID NO: 266).

[0428] - Payload

[0429] In this specification, the term "payload" refers to a substance that is bound to a fusion protein and transported together with the fusion protein. The payload may be, but is not limited to, a drug, a toxin, a fluorescent substance, an affinity ligand, a diagnostic substance, or a detection probe.

[0430] In one specific example, the payload may be a fluorescent substance. Examples of the fluorescent substance include, but are not limited to, FITC, GFP, YFP, CFP, IRDye® 800CW, and the like.

[0431] In one specific example, the payload may be a drug.

[0432] More specifically, the drug may be an anticancer agent.

[0433] As used herein, the term "anticancer agent" collectively refers to all drugs effective in treating cancer. In the present invention, the anticancer agent may include any anticancer agent that can be used as the payload of the fusion protein-drug conjugate of the present invention.

[0434] More specifically, it may be Camptothecin, Cryptophycins, Duocarmycin (including synthetic analogues, KW-2189 and CB1-TM1), Calicheamycin, Adrlimycin, Doxorubicin, Maytansinoids, Taxoids, Taxotrer, Doxetaxel, Chloranbucil, Gemcitabine, Platinum analog auristatin, Geldanamycin, Vindesine, Dolastatin, Dolastatin analog's auristatin, PBD (Pyrrolobenzodiazepine) derivatives, α-amanitin, β-amanitin, etc. In addition, it may include, but is not limited to, compounds that bind to tubulin binding, DNA binding, polymerase, topoisomerases, kinases, and exhibit cell growth inhibition and toxicity, immune function modulating drugs (e.g., TLR (Toll-like receptor) 7, TLR8 antagonists, STING antagonists, etc.), and targeted protein degraders (e.g., PROTAC (Proteolysis-targeting chimera)).

[0435] In one specific example, the anticancer agent may be MMAF or MMAE.

[0436] Table 8 below illustrates examples of linkers and drugs that may be included in drug conjugates provided herein. Fusion protein-drug conjugates according to exemplary embodiments of the present invention may include these seventh components. Furthermore, fusion protein-drug conjugates of the present invention are illustrated in Tables 44 and 45 as examples of the present invention.

[0437] Component 7: Linker-payload Part Number Linker-payload; UNIT 3 Variant Description G1 IRDye-800CWLI-COR, IRDye® 800CW Maleimide IRDye-800CW G2 LT-336 mal-FITCmal-FITCG3 LT-017 mal-VC-MMAFmal-VC-MMAFG4 LT-303 mal-TROSIG TM -MMAFmal-bGal-Thiophene2-MMAFG5LT-304mal-TROSIG TM -MMAFmal-bGal-Thiophene2'-MMAFG6LT-317mal-TROSIG TM -MMAF2mal-bGal-Se2'-MMAF2G7LT-327mal-TROSIG TM -MMAE2mal-bG-Se2'-MMAE2G8LT-348mal-TROSIG TM -MMAE2mal-bG-Se1''-MMAE2

[0438] multispecific fusion proteins

[0439] In the present invention, the fusion protein may additionally include a component selected from the group consisting of an Fc region or a variant thereof (fourth component), an antigen binding site (sixth component), a linker-payload (seventh component), and a combination thereof.

[0440] Specifically, the fusion protein according to the present invention may further include an Fc region or a variant thereof (fourth component) and / or an antigen binding site (seventh component), or an Fc region or a variant thereof (fourth component) and a linker-payload (seventh component) in ReleaDome, which is the minimum unit of the fusion protein of the present invention. In the present invention, the ReleaDome to which each of the above components is added is called a "multi-specific fusion protein." The ReleaDome and Each component is It can be connected via a linker. In this case, the linker used is the third linker, which is the same as described above.

[0441] The above multi-specific fusion protein may bind to at least one target or antigen.

[0442] Multispecific fusion protein containing a fourth component

[0443] In one specific example, the fusion protein may further comprise an Fc region or a variant thereof (fourth component). In this case, the fusion protein may be ReleaDome. ReleaDome is the same as described above.

[0444] In the present invention, the multi-antibody assembly rate, drug half-life (Pk), and effector functions of the multi-specific fusion proteins of the present invention can be controlled through the Fc region or a variant thereof, and multi-specific fusion proteins of various modality structures can be produced.

[0445] Hereinafter, the Fc region or its variant is described interchangeably with “fourth component” or “D”.

[0446] - Fc region and its variants

[0447] As used herein, the term "Fc region" refers to the Fc region of an immunoglobulin. The Fc region refers to a protein that includes the heavy chain constant region 2 (CH2) and the heavy chain constant region 3 (CH3) of an immunoglobulin, but does not include the variable regions of the heavy and light chains of the immunoglobulin, the heavy chain constant region 1 (CH1), and the light chain constant region (CL).

[0448] The Fc region of the above immunoglobulin may be a wild-type Fc domain or an Fc domain fragment. For example, the fragment of the Fc region may have lysine (K) deleted from the C-terminus. Alternatively, the fragment of the Fc region may contain only CH3. The immunoglobulin may be IgG, IgA, IgE, IgD, or IgM. Additionally, it may be IgG1, IgG2, IgG3, or IgG4, which are subclasses of IgG, or IgA1 or IgA2, which are subclasses of IgA.

[0449] As used herein, the term "Fc region variant" refers to a variant that has a different glycosylation pattern than the wild-type Fc region, or may have increased glycosylation compared to the wild-type Fc region, decreased glycosylation compared to the wild-type Fc region, or a deglycosylated form. An aglycosylated Fc region is also included. The Fc region or variant may have a modified number of sialic acid, fucosylation, or glycosylation through culture conditions or genetic manipulation of the host.

[0450] Additionally, the sugar chains of the Fc region of the immunoglobulin can be modified by conventional methods, such as chemical methods, enzymatic methods, and genetic engineering methods using microorganisms. In addition, the Fc domain variant may be a mixed form of the Fc regions of the immunoglobulin IgG, IgA, IgE, IgD, or IgM. In addition, the Fc region variant may be a form in which some amino acids of the Fc region are substituted.

[0451] The “amino acid” introduced by the above substitution and / or addition may be any one selected from the group consisting of lysine (K), alanine (A), arginine (R), asparagine (N), aspartic acid (D), cysteine ​​(C), glutamine (Q), glutamic acid (E), glycine (G), histidine (H), isoleucine (I), leucine (L), methionine (M), phenylalanine (F), proline (P), serine (S), threonine (T), tryptophan (W), tyrosine (Y), and valine (V).

[0452] Additionally, the Fc region variant may have a new linker introduced or a new domain substituted.

[0453] Additionally, the Fc region variant may include a knob structure or a hole structure.

[0454] "Knob-into-hole" is a design strategy for creating antibodies that bind specifically to different domains, such as bispecific antibodies, multispecific antibodies, or heterodimers. Generally, this technique involves introducing a knob at the interface of a first polypeptide chain (e.g., the first CH3 domain of a first antibody heavy chain) and a corresponding hole at the interface of a second polypeptide chain (e.g., the second CH3 domain of a second antibody heavy chain), such that the knob is positioned within the hole to promote heterodimer formation and prevent homodimer formation.

[0455] The 'knobs' are constructed by replacing small amino acid side chains from the interface of the first polypeptide (e.g., the first CH3 domain of the first antibody heavy chain) with larger side chains (e.g., arginine, phenylalanine, tyrosine, or tryptophan). Complementary 'holes' of the same or similar size to the knobs are created by replacing large amino acid side chains with smaller side chains (e.g., alanine, serine, valine, or threonine) at the interface of the second polypeptide (e.g., the second CH3 domain of the second antibody heavy chain). The knobs and holes can be created, for example, by site-specific mutagenesis of the nucleic acid encoding the polypeptide, or by alterations made during peptide synthesis.

[0456] Examples of knob-into-hole technology in the Fc region include mutations described in WO2014084607A1 and WO2018059502A1, etc. The disclosures of WO2014084607A1 and WO2018059502A1 are incorporated herein by reference. WO2014084607A1 discloses, for example, a binding of a tryptophan (W) substituted at Lys409 in one CH3 domain to a valine (V) substituted at Asp399 in another CH3 domain and a threonine (T) substituted at Phe405 in the CH3 domain (a-1); And (a-2) a binding of a serine (S) substituted at Tyr349 of one CH3 domain and a tryptophan (W) substituted at Glu357 of another CH3 domain, and further comprising (b-1) a binding of a glutamic acid (E) substituted at Lys360 of one CH3 domain and an arginine (R) substituted at Gln347 of another CH3 domain; and (b-2) a binding of a glutamic acid (E) substituted at Gln347 of one CH3 domain and a glutamic acid substituted at Lys360 and an arginine (R) substituted at Gln347 of another CH3 domain. Herein, the positions of the amino acid residues are according to the EU index. WO2018059502A1 describes, for example, a mutation of the Fc domain comprising one or more mutations selected from each of the following a)-e): a) L351G, L351Y, L351V, L351P, L351D, L351E, L351K, or L351W; b) T366L, T366P, T366W, or T366V; c) D399C, D399N, D399I, D399G, D399R, D399T, or D399A; d) Y407L, Y407A, Y407P, Y407F, Y407T, or Y407H; and e) K409C, K409P, K409S, K409F, K409V, K409Q, or K409R.Here, the positions of the amino acid residues are according to the EU index.

[0457] Table 9 below shows the amino acid sequences of the Fc regions or variants thereof of various fusion proteins described herein. Fusion proteins according to exemplary embodiments may include an Fc region or variant thereof comprising or consisting of these. The amino acid sequences below are listed sequentially from the N-terminus to the C-terminus.

[0458]

[0459]

[0460] The above fusion protein and the Fc region or variant thereof may be linked via a linker. In this case, the fusion protein may be ReleaDome. ReleaDome is the same as described above.

[0461] Specifically, the Fc region or a variant thereof may be linked to the C-terminus of the first construct via a linker.

[0462] More specifically, it may include the following structural formulas (I') and (II).

[0463] N'-T1'-((L1)a-CR1)b-(L2')-I1'-((L3)c-CR3)d-(L5)-Fc region or a variant thereof-C' (I'); and

[0464] N'-T1''-((L1)a-CR2)b-(L2'')-I1''-((L3)c-CR4)dC' (II)

[0465] At this time, in the structural formulas (I') and (II),

[0466] The above N' is the N-terminus of the fusion protein,

[0467] The above C' is the C-terminus of the fusion protein,

[0468] The above T1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the first target,

[0469] The above T1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the first target,

[0470] The above T1' and T1'' combine to form a binding site (T1) that specifically binds to the first target,

[0471] The above I1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target,

[0472] The above I1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target,

[0473] The above I1' and I1'' combine to form a binding site (I1) that specifically binds to the second target,

[0474] The above CR1, CR2, CR3 and CR4 are each independently CH1 or CL,

[0475] At this time, CR1 and CR2 are not equal, CR3 and CR4 are not equal,

[0476] The above CH1 is a heavy chain constant region,

[0477] The above CL is a light chain constant region,

[0478] The above L1, L2', L2'', L3 and L5 are peptide linkers, wherein L2' and L2'' are cleavable or non-cleavable linkers,

[0479] The above a, b, c and d are each independently 0 or 1.

[0480] The first target, second target, heavy chain variable region, light chain variable region, heavy chain constant region, light chain constant region, Fc region and variants thereof are the same as described above.

[0481] The above L1 and L3 may be first-type non-cleavable linkers, and L2' and L2'' may be first linkers. The first-type non-cleavable linker, the first linker, the second linker, and the third linker are the same as described above.

[0482] The combination of the sixth structure and the second structure of the above structural formula (I') can be achieved through i) a disulfide bond by cysteine ​​(Cys) present in the heavy chain constant region (CH1) and the light chain constant region (CL) of CR3 and CR4, ii) a disulfide bond by cysteine ​​(Cys) present in the heavy chain variable region (VH2) and the light chain variable region (VL2) within I1' and I1'', or iii) a disulfide bond by cysteine ​​(Cys) present in the heavy chain constant region (CH1) and the light chain constant region (CL) of CR3 and CR4 and a disulfide bond by cysteine ​​(Cys) present in the heavy chain variable region (VH2) and the light chain variable region (VL2) within I1' and I1''.

[0483] Specifically, the Fc region or a variant thereof may be linked to the C-terminus of the fourth structure via a linker.

[0484] More specifically, it may include the following structural formulas (IV') and (V).

[0485] N'-T2-(L7)-I2'-((L8)f-CR1)g-(L9)-Fc region or a variant thereof-C' (IV');

[0486] N'-I2''-((L8)f-CR2)hC' (V)

[0487] At this time, in the structural formulas (IV') and (V),

[0488] The above N' is the N-terminus of the fusion protein,

[0489] The above C' is the C-terminus of the fusion protein,

[0490] The above T2 is a peptide or antibody fragment that specifically binds to the first target,

[0491] The above I2' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target,

[0492] The above I2'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target,

[0493] The above I2' and I2'' combine to form a binding site (I2) that specifically binds to the second target,

[0494] The above CR1 and CR2 are each independently CH1 or CL,

[0495] At this time, CR1 and CR2 are not equal,

[0496] The above CH1 is a heavy chain constant region,

[0497] The above CL is a light chain constant region,

[0498] The above L7, L8 and L9 are each independently a peptide linker, wherein L7 is a cleavable or non-cleavable linker,

[0499] The above f, g and h are each independently 0 or 1, and wherein g and h are the same.

[0500] The first target, second target, heavy chain variable region, light chain variable region, heavy chain constant region, light chain constant region, and Fc region and variants thereof are the same as described above.

[0501] The above L7 may be a first linker, L8 may be a first-type non-cleavable linker, and L9 may be a third linker. The first-type non-cleavable linker, the first linker, and the third linker are the same as described above.

[0502] The bonding of the seventh structure and the fifth structure of the above structural formula (IV') can be formed through i) a disulfide bond by cysteine ​​(Cys) present in the heavy chain constant region (CH1) and the light chain constant region (CL), ii) a disulfide bond by cysteine ​​(Cys) present in the heavy chain variable region (VH2) and the light chain variable region (VL2) within I2' and I2'', or iii) a disulfide bond by cysteine ​​(Cys) present in the heavy chain constant region (CH1) and the light chain constant region (CL) and a disulfide bond by cysteine ​​(Cys) present in the heavy chain variable region (VH2) and the light chain variable region (VL2) within I2' and I2''.

[0503] In the present invention, the multi-specific fusion protein comprising the sixth or seventh structure may additionally comprise a payload. The payload may be linked to a specific or unspecified position of the fourth component, the Fc region or a variant thereof, in one embodiment of the multi-specific fusion protein of the present invention, to form a functional entity of the fusion protein-drug conjugate. In this case, the Fc region or the variant thereof and the payload may be linked via a linker. The linker may be a second linker.

[0504] The above multi-specific fusion proteins and payloads can be conjugated using known technologies. For example, the lysine (K) and AcBut-calicheamicin linkage technology used in Mylotarg® and Besponsa®, the lysine (K) and SMCC-DM1 linkage technology used in Kadcyla®, and the thiol-maleimide technology used in Adcetris®, Polivy®, Padcev®, Enhertu®, Trodelvy®, and Blenrep®, but are not limited thereto. In addition, THIOMAB using modified cysteine ​​is also used. TM Technology, C-Lock using Dibromomethylquinoxaline functional group TM, SMARTag™ using Pictet-Spengler reaction, GlycoConnect™ using Azidosugar, ConjuAll™ using Prenyltransferase, SMAC-Technology™ using Sortase, and glutamine coupling technology using Transglutaminase, but are not limited thereto.

[0505] In addition, the multispecific fusion protein comprising the sixth or seventh structure may further comprise a peptide motif. In this case, the peptide motif may be linked to the C-terminus of the Fc region of the sixth or seventh structure or a variant thereof via a linker. When the multispecific fusion protein comprising the sixth or seventh structure comprises a peptide motif, the seventh component may be linked to the peptide motif through an enzymatic reaction. In one specific example, the peptide motif may comprise a CAAX motif, and more specifically, may comprise the amino acid sequence CVIM (SEQ ID NO: 267). In this case, the first type non-cleavable linker may be GGGGGGG (SEQ ID NO: 266). The payload, linker, peptide motif, and seventh component are the same as described above.

[0506] Multispecific fusion protein comprising the fourth and sixth components

[0507] In the present invention, the fusion protein may additionally include an Fc region or a variant thereof (fourth component) and an antigen binding site (sixth component).

[0508] Specifically, the antigen binding site may be linked to the C-terminus of the sixth structure via a linker.

[0509] More specifically, it may include the following structural formulas (I'') and (II).

[0510] N'-T1'-((L1)a-CR1)b-(L2')-I1'-((L3)c-CR3)d-(L5)-Fc region or a variant thereof-(L6)eOC' (I''); and

[0511] N'-T1''-((L1)a-CR2)b-(L2'')-I1''-((L3)c-CR4)dC' (II)

[0512] At this time, in the structural formulas (I'') and (II),

[0513] The above N' is the N-terminus of the fusion protein,

[0514] The above C' is the C-terminus of the fusion protein,

[0515] The above T1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the first target,

[0516] The above T1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the first target,

[0517] The above T1' and T1'' combine to form a binding site (T1) that specifically binds to the first target,

[0518] The above I1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target,

[0519] The above I1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target,

[0520] The above I1' and I1'' combine to form a binding site (I1) that specifically binds to the second target,

[0521] The above CR1, CR2, CR3 and CR4 are each independently CH1 or CL,

[0522] At this time, CR1 and CR2 are not equal, CR3 and CR4 are not equal,

[0523] The above CH1 is a heavy chain constant region,

[0524] The above CL is a light chain constant region,

[0525] The above O is an antigen binding site,

[0526] The above L1, L2', L2'', L3, L5 and L6 are each independently a peptide linker, wherein L2' and L2'' are cleavable or non-cleavable linkers,

[0527] The above a, b, c, d and e are each independently 0 or 1.

[0528] The first target, second target, heavy chain variable region, light chain variable region, heavy chain constant region, light chain constant region, and Fc region and variants thereof are the same as described above.

[0529] The above L1, L3, and L6 are first linkers, specifically, type 1 non-cleavable linkers, and L2' and L2'' may be first linkers. The above L5 may be a third linker. The type 1 non-cleavable linker, the first linker, the second linker, and the third linker are the same as described above.

[0530] At this time, the combination of the 8th structure of the structural formula (I'') and the 2nd structure can be formed through i) a disulfide bond by cysteine ​​(Cys) present in the heavy chain constant region (CH1) and the light chain constant region (CL) of CR3 and CR4, ii) a disulfide bond by cysteine ​​(Cys) present in the heavy chain variable region (VH2) and the light chain variable region (VL2) within I1' and I1'', or iii) a disulfide bond by cysteine ​​(Cys) present in the heavy chain constant region (CH1) and the light chain constant region (CL) of CR3 and CR4 and a disulfide bond by cysteine ​​(Cys) present in the heavy chain variable region (VH2) and the light chain variable region (VL2) within I1' and I1''.

[0531] Specifically, the fusion protein may have an antigen binding site linked to the C-terminus of the seventh structure via a linker.

[0532] More specifically, it may include the following structural formulas (IV'') and (V).

[0533] N'-T2-(L7)-I2'-((L8)f-CR1)g-(L9)-Fc region or a variant thereof-(L10)iPC' (IV'');

[0534] N'-I2''-((L8)f-CR2)hC' (V)

[0535] At this time, in the structural formulas (IV'') and (V),

[0536] The above N' is the N-terminus of the fusion protein,

[0537] The above C' is the C-terminus of the fusion protein,

[0538] The above T2 is a peptide or antibody fragment that specifically binds to the first target,

[0539] The above I2' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target,

[0540] The above I2'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target,

[0541] The above I2' and I2'' combine to form a binding site (I2) that specifically binds to the second target,

[0542] The above CR1 and CR2 are each independently CH1 or CL,

[0543] At this time, CR1 and CR2 are not equal,

[0544] The above CH1 is a heavy chain constant region,

[0545] The above CL is a light chain constant region,

[0546] The above P is an antigen binding site,

[0547] The above L7, L8, L9 and L10 are each independently a peptide linker, wherein L7 is a cleavable or non-cleavable linker,

[0548] The above f, g, h and i are each independently 0 or 1.

[0549] The first target, second target, heavy chain variable region, light chain variable region, heavy chain constant region, light chain constant region and Fc region and variants thereof are the same as described above.

[0550] The above L7 may be a first linker, L8 and L10 may be type 1 non-cleavable linkers, and L9 may be a third linker. The type 1 non-cleavable linker, the first linker, and the third linker are the same as described above.

[0551] At this time, the combination of the 9th structure and the 5th structure of the structural formula (IV'') may be formed through i) a disulfide bond by cysteine ​​(Cys) present in the heavy chain constant region (CH1) and the light chain constant region (CL), ii) a disulfide bond by cysteine ​​(Cys) present in the heavy chain variable region (VH2) and the light chain variable region (VL2) within I2' and I2'', or iii) a disulfide bond by cysteine ​​(Cys) present in the heavy chain constant region (CH1) and the light chain constant region (CL) and a disulfide bond by cysteine ​​(Cys) present in the heavy chain variable region (VH) and the light chain variable region (VL) within I2' and I2''.

[0552] - Antigen binding site

[0553] The antigen binding site may be an antigen binding site that specifically binds to a protein that is overexpressed, specifically expressed, or activated in target cells at the disease site. The protein that is overexpressed, specifically expressed, or activated in target cells at the disease site is the same as described above. The antigen may be the same as or different from the first target and / or the second target.

[0554] As an example of the above antigen binding site, it may be an antibody or a fragment thereof (e.g., Fv, scFv, VHH, Fab), a peptide, an aptamer, etc., and may be in a form that includes part or all of the counterpart / ligand existing in nature of the target that can be the antigen of the second domain, but is not limited thereto. More specifically, it may be an antibody or a fragment thereof. The antibody and fragment thereof are the same as described above. More preferably, it may be an scFv.

[0555] Additionally, examples of the antigen may include, but are not limited to, CLDN18.2 or PD-L1.

[0556] In one specific example, the antigen binding site may include a heavy chain variable region and a light chain variable region of Zolbetuximab, an antibody that binds to CLDN18.2. Specifically, the antigen binding site may include a heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 872, HCDR2 comprising the amino acid sequence of SEQ ID NO: 874, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 876, and a light chain variable region comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 968, LCDR2 comprising the amino acid sequence of SEQ ID NO: 559, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 971. In this case, the heavy chain variable region may include the amino acid sequence of SEQ ID NO: 222, and the light chain variable region may include the amino acid sequence of SEQ ID NO: 223. The antigen binding site may include the amino acid sequence of SEQ ID NO: 268.

[0557] In one specific example, the antigen binding site may include a heavy chain variable region and a light chain variable region of Atezolizumab, an antibody that binds to PD-L1. Specifically, the antigen binding site may include a heavy chain variable region comprising HCDR1 comprising an amino acid sequence of SEQ ID NO: 895, HCDR2 comprising an amino acid sequence of SEQ ID NO: 898, and HCDR3 comprising an amino acid sequence of SEQ ID NO: 900, and a light chain variable region comprising LCDR1 comprising an amino acid sequence of SEQ ID NO: 982, LCDR2 comprising an amino acid sequence of SEQ ID NO: 680, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 984. In this case, the heavy chain variable region may include an amino acid sequence of SEQ ID NO: 230, and the light chain variable region may include an amino acid sequence of SEQ ID NO: 231. The antigen binding site may include an amino acid sequence of SEQ ID NO: 238.

[0558] Table 10 below shows the polypeptide sequences of antibody fragments described as antigen-binding sites (sixth component) of various multispecific fusion proteins described herein. Fusion proteins according to exemplary embodiments may include these polypeptides. The polypeptide sequences are listed sequentially from the N-terminus to the C-terminus.

[0559] In this specification, the antigen binding site of the sixth component is referred to as “UNIT2’”, and hereinafter, the antigen binding site is described interchangeably with “sixth component” or “F”.

[0560]

[0561]

[0562] Multispecific fusion protein containing the sixth component

[0563] In the present invention, the fusion protein may additionally include an antigen binding site (sixth component). In this case, the fusion protein may be ReleaDome. The fusion protein and the antigen binding site may be connected via a linker, and the linker may be a third linker. In addition, the antigen binding site may be a variable region of an antibody, and specifically, may be a heavy chain variable region or a light chain variable region of an antibody. The third linker, antibody, variable region, heavy chain variable region, and light chain variable region are the same as described above.

[0564] Specifically, the antigen binding site can be linked to the variable region of the antibody via a third linker at the C-terminus of the first structure.

[0565] More specifically, it may include the following structural formulas (I''') and (II).

[0566] N'-T1'-((L1)a-CR1)b-(L2')-I1'-((L3)c-CR3)d-(L5)-(L18)s-Q'-C' (I'''); and

[0567] N'-T1''-((L1)a-CR2)b-(L2'')-I1''-((L3)c-CR4)dC' (II)

[0568] At this time, in the structural formulas (I''') and (II),

[0569] The above N' is the N-terminus of the fusion protein,

[0570] The above C' is the C-terminus of the fusion protein,

[0571] The above T1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the first target,

[0572] The above T1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the first target,

[0573] The above T1' and T1'' combine to form a binding site (T1) that specifically binds to the first target,

[0574] The above I1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target,

[0575] The above I1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target,

[0576] The above I1' and I1'' combine to form a binding site (I1) that specifically binds to the second target,

[0577] The above CR1, CR2, CR3 and CR4 are each independently CH1 or CL,

[0578] At this time, CR1 and CR2 are not equal, CR3 and CR4 are not equal,

[0579] The above CH1 is a heavy chain constant region,

[0580] The above CL is a light chain constant region,

[0581] The above Q' is a heavy chain variable region (VH) of an antibody that specifically binds to the third target,

[0582] The above L1, L2', L2'', L3, L5 and L18 are each independently a peptide linker, wherein L2 is a cleavable or non-cleavable linker,

[0583] The above a, b, c, d and s are each independently 0 or 1, and b and d are the same.

[0584] Alternatively, it may include the following structural formulas (I'''') and (II).

[0585] N'-T1'-((L1)a-CR1)b-(L2')-I1'-((L3)c-CR3)d-(L5)-(L18)s-Q''-C' (I''''); and

[0586] N'-T1''-((L1)a-CR2)b-(L2'')-I1''-((L3)c-CR4)dC' (II)

[0587] At this time, in the structural formulas (I'''') and (II),

[0588] The above N' is the N-terminus of the fusion protein,

[0589] The above C' is the C-terminus of the fusion protein,

[0590] The above T1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the first target,

[0591] The above T1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the first target,

[0592] The above T1' and T1'' combine to form a binding site (T1) that specifically binds to the first target,

[0593] The above I1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target,

[0594] The above I1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target,

[0595] The above I1' and I1'' combine to form a binding site (I1) that specifically binds to the second target,

[0596] The above CR1, CR2, CR3 and CR4 are each independently CH1 or CL,

[0597] At this time, CR1 and CR2 are not equal, CR3 and CR4 are not equal,

[0598] The above CH1 is a heavy chain constant region,

[0599] The above CL is a light chain constant region,

[0600] The above Q'' is a light chain variable region (VL) of an antibody that specifically binds to the third target,

[0601] The above L1, L2', L2'', L3, L5 and L18 are each independently a peptide linker, wherein L2' and L2'' are cleavable or non-cleavable linkers,

[0602] The above a, b, c, d and s are each independently 0 or 1, and b and d are the same.

[0603] The first target, second target, heavy chain variable region, light chain variable region, heavy chain constant region, and light chain constant region are the same as described above.

[0604] The heavy chain variable region and light chain variable region Q' and Q'' of the antibody that specifically binds to the third target can combine to form an antigen binding site that binds to the same antigen (Q, the third target). The antigen may be a protein that is overexpressed or specifically expressed or activated in target cells at the disease site. The protein that is overexpressed or specifically expressed or activated in target cells at the disease site is the same as described above. In addition, the third target may be the same as or different from the first target and / or the second target.

[0605] The above L1, L3 and L18 may each independently be a type 1 non-cleavable linker, and L2' and L2'' may be a type 1 non-cleavable linker or a type 1 cleavable linker. In addition, the above L5 may be a third linker.

[0606] The protein, the first linker, the type 1 cleavable linker, the type 1 non-cleavable linker, and the third linker that are overexpressed or specifically expressed or activated in the target cells at the disease site are the same as described above.

[0607] The bonding of the eleventh structure of the structural formula (I''') or the twelfth structure of the structural formula (I'''') and the second structure may be formed i) through a disulfide bond by cysteine ​​(Cys) present in the heavy chain constant region (CH1) and the light chain constant region (CL) of CR3 and CR4, ii) through a disulfide bond by cysteine ​​(Cys) present in the heavy chain variable region (VH2) and the light chain variable region (VL2) within I' and I'', or iii) through a disulfide bond by cysteine ​​(Cys) present in the heavy chain constant region (CH1) and the light chain constant region (CL) of CR3 and CR4 and a disulfide bond by cysteine ​​(Cys) present in the heavy chain variable region (VH2) and the light chain variable region (VL2) within I1' and I1''.

[0608] The above antigen binding site can be linked to the variable region of the antibody via a third linker at the C-terminus of the fourth structure.

[0609] Specifically, it may include the following structural formulas (IV''') and (V).

[0610] N'-T2-(L7)-I2'-((L8)f-CR1)g-(L9)-(L19)t-S'-C' (IV''');

[0611] N'-I2''-((L8)f-CR2)hC' (V)

[0612] At this time, in the structural formulas (IV''') and (V),

[0613] The above N' is the N-terminus of the fusion protein,

[0614] The above C' is the C-terminus of the fusion protein,

[0615] The above T2 is a peptide or antibody fragment that specifically binds to the first target,

[0616] The above I2' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target,

[0617] The above I2'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target,

[0618] The above I2' and I2'' combine to form a binding site (I2) that specifically binds to the second target,

[0619] The above CR1 and CR2 are each independently CH1 or CL,

[0620] At this time, CR1 and CR2 are not equal,

[0621] The above CH1 is a heavy chain constant region,

[0622] The above CL is a light chain constant region,

[0623] The above S' is a heavy chain variable region (VH) of an antibody that specifically binds to the fourth target,

[0624] The above L7, L8, L9 and L19 are each independently a peptide linker, wherein L7 is a cleavable or non-cleavable linker,

[0625] The above f, g, h and t are each independently 0 or 1, and wherein g and h are the same.

[0626] Alternatively, it may include the following structural formulas (IV'''') and (V).

[0627] N'-T2-(L7)-I2'-((L8)f-CR1)g-(L10)-(L19)t-S''-C' (IV'''');

[0628] N'-I2''-((L8)f-CR2)hC' (V)

[0629] At this time, in the structural formulas (IV'''') and (V),

[0630] The above N' is the N-terminus of the fusion protein,

[0631] The above C' is the C-terminus of the fusion protein,

[0632] The above T2 is a peptide or antibody fragment that specifically binds to the first target,

[0633] The above I2' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target,

[0634] The above I2'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target,

[0635] The above I2' and I2'' combine to form a binding site (I2) that specifically binds to the second target,

[0636] The above CR1 and CR2 are each independently CH1 or CL,

[0637] At this time, CR1 and CR2 are not equal,

[0638] The above CH1 is a heavy chain constant region,

[0639] The above CL is a light chain constant region,

[0640] The above S'' is a light chain variable region (VL) of an antibody that specifically binds to the fourth target,

[0641] The above L7, L8, L10 and L19 are each independently a peptide linker, wherein L7 is a cleavable or non-cleavable linker,

[0642] The above f, g, h and t are each independently 0 or 1, and wherein g and h are the same.

[0643] The first target, second target, heavy chain variable region, light chain variable region, heavy chain constant region, and light chain constant region are the same as described above.

[0644] The heavy chain variable region and light chain variable region, S' and S'' of the above antibody, can combine to form an antigen binding site that binds to the same antigen (S, the fourth target). The antigen may be a protein that is overexpressed or specifically expressed or activated in target cells at the site of the disease. The antigen is the same as the protein that is overexpressed or specifically expressed or activated in target cells at the site of the disease. In addition, the fourth target may be the same as or different from the first target and / or the second target.

[0645] The above L7 may be a first linker, L8 may be a first-type non-cleavable linker, and L10 may be a third linker.

[0646] The protein, the first linker, the type 1 cleavable linker, the type 1 non-cleavable linker, and the third linker that are overexpressed or specifically expressed or activated in the target cells at the disease site are the same as described above.

[0647] The bonding of the 13th structure of the above structural formula (IV''') or the 14th structure and the 5th structure of the structural formula (IV'''') can be formed through i) a disulfide bond by cysteine ​​(Cys) present in the heavy chain constant region (CH1) and the light chain constant region (CL), ii) a disulfide bond by cysteine ​​(Cys) present in the heavy chain variable region (VH) and the light chain variable region (VL) within I2' and I2'', or iii) a disulfide bond by cysteine ​​(Cys) present in the heavy chain constant region (CH1) and the light chain constant region (CL) and a disulfide bond by cysteine ​​(Cys) present in the heavy chain variable region (VH2) and the light chain variable region (VL2) within I2' and I2''.

[0648] Fusion protein dimer

[0649] Another aspect of the present invention provides a fusion protein dimer comprising two of the above fusion proteins.

[0650] At this time, the fusion protein may be a multi-specific fusion protein. The multi-specific fusion protein is the same as described above.

[0651] multispecific fusion protein homodimer

[0652] In the present invention, the fusion protein dimer may be a homodimer.

[0653] In one specific example, the fusion protein dimer may be a combination of two identical multispecific fusion proteins comprising a sixth structure and a second structure. In one specific example, the fusion protein dimer may be a combination of two identical multispecific fusion proteins comprising a seventh structure and a fifth structure. In one specific example, the fusion protein dimer may be a combination of two identical multispecific fusion proteins comprising an eighth structure and a second structure. In one specific example, the fusion protein dimer may be a combination of two identical multispecific fusion proteins comprising a ninth structure and a fifth structure. The second structure, the fifth structure to the ninth structure are the same as described above.

[0654] multispecific fusion protein heterodimer

[0655] The above fusion protein dimer may be a heterodimer.

[0656] In one specific embodiment, the fusion protein dimer may be a combination of two different multispecific fusion proteins comprising a sixth structure and a second structure. In one specific embodiment, the fusion protein dimer may be a combination of two different multispecific fusion proteins comprising a seventh structure and a fifth structure. In one specific embodiment, the fusion protein dimer may be a combination of two different multispecific fusion proteins comprising an eighth structure and a second structure. In one specific embodiment, the fusion protein dimer may be a combination of two different multispecific fusion proteins comprising a ninth structure and a fifth structure.

[0657] In addition, the fusion protein dimer may be a combination of a multi-specific fusion protein comprising the sixth structure and the second structure, or a multi-specific fusion protein comprising the eighth structure and the second structure; and a multi-specific fusion protein comprising the seventh structure and the fifth structure, or a multi-specific fusion protein comprising the ninth structure and the fifth structure. In this case, the first targets of the structures may be the same or different, and the second targets may be the same or different. The second structure, the fifth structure, and the ninth structure are the same as described above.

[0658] In one specific example, the fusion protein dimer may be a combination of a multispecific fusion protein comprising the sixth structure and the second structure; and a multispecific fusion protein comprising the seventh structure and the fifth structure. In one specific example, the fusion protein dimer may be a combination of a multispecific fusion protein comprising the sixth structure and the second structure; and a multispecific fusion protein comprising the ninth structure and the fifth structure. The fusion protein dimer may be a combination of a multispecific fusion protein comprising the eighth structure and the second structure; and a multispecific fusion protein comprising the seventh structure and the fifth structure. In one specific example, the fusion protein dimer may be a combination of a multispecific fusion protein comprising the eighth structure and the second structure; and a multispecific fusion protein comprising the ninth structure and the fifth structure.

[0659] Specifically, the fusion protein dimer may be a combination of a multispecific fusion protein comprising the 11th structure and the 2nd structure, or a multispecific fusion protein comprising the 13th structure and the 5th structure; and a multispecific fusion protein comprising the 12th structure and the 2nd structure, or a multispecific fusion protein comprising the 14th structure and the 5th structure. At this time, the first targets of the structures may be the same or different, and the second targets may be the same or different, respectively. The heavy chain variable region and the light chain variable region of the antibody, which are the antigen binding sites, may combine to form a binding site that binds to the same antigen. The second structure, the 5th structure, and the 11th to 14th structures are the same as described above.

[0660] In one specific example, the fusion protein dimer may be a combination of a multispecific fusion protein comprising the eleventh structure and the second structure; and a multispecific fusion protein comprising the twelfth structure and the second structure. In one specific example, the fusion protein dimer may be a combination of a multispecific fusion protein comprising the eleventh structure and the second structure; and a multispecific fusion protein comprising the fourteenth polystructure and the fifth structure. In one specific example, the fusion protein dimer may be a combination of a multispecific fusion protein comprising the thirteenth structure and the fifth structure; and a multispecific fusion protein comprising the twelfth structure and the second structure. In one specific example, the fusion protein dimer may be a combination of a multispecific fusion protein comprising the thirteenth structure and the fifth structure; and a multispecific fusion protein comprising the fourteenth structure and the fifth structure.

[0661] Specifically, the fusion protein dimer may be a combination of the multi-specific fusion protein and a fusion protein comprising the following structural formulas (VI) and (VII).

[0662] N'-I3'-(L11)k-CR1-(L12)-Fc region or a variant thereof-((L13)lV)mC' (VI); and

[0663] N'-I3''-(L11)k-CR2-C' (VII)

[0664] At this time, in the structural formulas (VI) and (VII),

[0665] The above N' is the N-terminus of the fusion protein,

[0666] The above C' is the C-terminus of the fusion protein,

[0667] The above I3' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target,

[0668] The above I3'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target,

[0669] The above I3' and I3'' form a binding site (I3) that specifically binds to the second target,

[0670] The above CR1 and CR2 are each independently CH1 or CL,

[0671] At this time, CR1 and CR2 are not the same,

[0672] The above CH1 is a heavy chain constant region,

[0673] The above CL is a light chain constant region,

[0674] The above V is an antigen binding site,

[0675] The above L11, L12 and L13 are each independently a peptide linker,

[0676] The above k, l and m are each independently 0 or 1.

[0677] The above L11 and L13 may be a first-type non-cleavable linker, and L12 may be a third linker. In addition, the second target and antigen binding site in the structural formulas (VI) and (VII) may be the same as or different from the second target and antigen binding site of the multispecific fusion protein, respectively. The second target, the antigen binding site, the antibody fragment, the first-type non-cleavable linker, and the third linker are the same as described above.

[0678] Hereinafter, in the present specification, the structure of the structural formula (VI) is described as the 15th structure, and the structure of the structural formula (VII) is described as the 16th structure. At this time, the fusion protein including the 15th structure may additionally include a payload. The payload may be connected via a linker, and the linker may be a second linker. In addition, the fusion protein including the 15th structure and the 16th structure may additionally include a peptide motif. At this time, the peptide motif may be connected via a linker to the C-terminus of the Fc region of the 15th structure or a variant thereof; or to the C-terminus of the light chain constant region (CL) of the 16th structure. When the fusion protein including the 15th structure or the 16th structure includes a peptide motif, the seventh component may be bound to the peptide motif through an enzymatic reaction. In one specific example, the peptide motif may comprise a CAAX motif, and more specifically, may comprise the amino acid sequence CVIM (SEQ ID NO: 267). In this case, the first type non-cleavable linker may be GGGGGGG (SEQ ID NO: 266). The payload, the second linker, the linker, the peptide motif, and the seventh component are the same as described above.

[0679] More specifically, the fusion protein dimer may be a combination of any one multispecific fusion protein selected from the group consisting of a multispecific fusion protein comprising the sixth structure and the second structure, a multispecific fusion protein comprising the eighth structure and the second structure, a multispecific fusion protein comprising the seventh structure and the fifth structure, and a multispecific fusion protein comprising the ninth structure and the fifth structure; and a fusion protein comprising the fifteenth structure and the sixteenth structure.

[0680] In one specific example, the fusion protein dimer may be a combination of a multispecific fusion protein comprising the sixth structure and the second structure; and a fusion protein comprising the fifteenth structure and the sixteenth structure. In one specific example, the fusion protein dimer may be a combination of a multispecific fusion protein comprising the eighth structure and the second structure; and a fusion protein comprising the fifteenth structure and the sixteenth structure. In one specific example, the fusion protein dimer may be a combination of a multispecific fusion protein comprising the seventh structure and the fifth structure; and a fusion protein comprising the fifteenth structure and the sixteenth structure. In one specific example, the fusion protein dimer may be a combination of a multispecific fusion protein comprising the ninth structure and the fifth structure; and a fusion protein comprising the fifteenth structure and the sixteenth structure.

[0681] Specifically, the fusion protein dimer may be a combination of the multi-specific fusion protein and a fusion protein comprising the following structural formula (VIII).

[0682] N'-I3'''-(L14)-Fc region or its variant-((L15)nW)oC' (VIII)

[0683] At this time, in the above structural formula (VIII),

[0684] The above N' is the N-terminus of the fusion protein,

[0685] The above C' is the C-terminus of the fusion protein,

[0686] The above I3''' is an antibody fragment that specifically binds to the second target,

[0687] The above W is an antigen binding site,

[0688] The above L14 and L15 are each independently a peptide linker,

[0689] The above n and o are each independently 0 or 1.

[0690] The above L14 may be a third linker, and L15 may be a type 1 non-cleavable linker.

[0691] Additionally, in the structural formula (VIII), the second target and antigen binding sites may be identical to or different from the second target and antigen binding sites of the multispecific fusion protein, respectively. The second target, antigen binding site, antibody fragment, type 1 cleavable linker, and third linker are identical to those described above.

[0692] Hereinafter, in the present specification, the structure of the structural formula (VIII) is described as a third structure. At this time, the fusion protein including the third structure may additionally include a payload. The payload may be connected via a linker, and the linker may be a third linker. In addition, the fusion protein including the third structure may additionally include a peptide motif. At this time, the peptide motif may be connected to the C-terminus of the Fc region of the third structure or a variant thereof via a linker. When the fusion protein including the third structure includes a peptide motif, the seventh component may be bound to the peptide motif through an enzymatic reaction. In one specific example, the peptide motif may include a CAAX motif, and more specifically, may include the amino acid sequence CVIM (SEQ ID NO: 267). At this time, the first type non-cleavable linker may be GGGGGGG (SEQ ID NO: 266). The payload, the third linker, the linker, the peptide motif, and the seventh component are the same as described above.

[0693] More specifically, the fusion protein dimer may be a combination of any one multispecific fusion protein selected from the group consisting of a multispecific fusion protein comprising the sixth structure and the second structure, a multispecific fusion protein comprising the eighth structure and the second structure, a multispecific fusion protein comprising the seventh structure and the fifth structure, and a multispecific fusion protein comprising the ninth structure and the fifth structure; and a fusion protein comprising the third structure.

[0694] In one specific example, the fusion protein dimer may be a combination of a multispecific fusion protein comprising the sixth structure and the second structure; and a fusion protein comprising the third structure. In one specific example, the fusion protein dimer may be a combination of a multispecific fusion protein comprising the eighth structure and the second structure; and a fusion protein comprising the third structure. In one specific example, the fusion protein dimer may be a combination of a multispecific fusion protein comprising the seventh structure and the fifth structure; and a fusion protein comprising the third structure. In one specific example, the fusion protein dimer may be a combination of a multispecific fusion protein comprising the ninth structure and the fifth structure; and a fusion protein comprising the third structure.

[0695] Specific examples of fusion protein dimers

[0696] - Multispecific fusion protein dimer in the form of DVD-Ig containing ReleaDome

[0697] As shown in Fig. 3a, the multispecific fusion protein dimer in the form of a DVD-Ig including ReleaDome may be a homodimer in which two multispecific fusion proteins are combined, the sixth structure of structural formula (I') and the second structure of structural formula (II).

[0698] Specifically, the sixth structure may be a multi-specific fusion protein comprising a first domain (first component) comprising a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody, and a second domain (third component) comprising a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2) and a light chain constant region (CL) of an antibody, which are connected in series via a first linker (second component), and a ReleaDome linked to an Fc region or a variant thereof (fourth component) via a third linker. In addition, the CL may further comprise a type 1 non-cleavable protein and a peptide motif.

[0699] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-ANXA1-anti-HER2]

[0700] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody that binds to ANXA1, and a second domain (third component) including a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2), and a light chain constant region (CL) of an antibody that binds to HER2 are serially connected through a first linker (second component), and an Fc region or a variant thereof (fourth component) is connected through a third linker.

[0701] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, wherein a sixth structure comprises a heavy chain variable region (VH1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to HER2, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprises a light chain variable region (VL1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to HER2, a type 1 non-cleavable linker, and a light chain constant region (CL).

[0702] In one specific example, it may include a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 991 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 992. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 993 and a light chain comprising the amino acid sequence of SEQ ID NO: 994. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 995 and a light chain comprising the amino acid sequence of SEQ ID NO: 996. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 997 and a light chain comprising the amino acid sequence of SEQ ID NO: 998. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 999 and a light chain comprising the amino acid sequence of SEQ ID NO: 1000. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1001 and a light chain comprising the amino acid sequence of SEQ ID NO: 1002. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1003 and a light chain comprising the amino acid sequence of SEQ ID NO: 1004. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1005 and a light chain comprising the amino acid sequence of SEQ ID NO: 1006. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1007 and a light chain comprising the amino acid sequence of SEQ ID NO: 1008. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1009 and a light chain comprising the amino acid sequence of SEQ ID NO: 1010. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1011 and a light chain comprising the amino acid sequence of SEQ ID NO: 1012. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1013 and a light chain comprising the amino acid sequence of SEQ ID NO: 1014.In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1015 and a light chain comprising the amino acid sequence of SEQ ID NO: 1016. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1017 and a light chain comprising the amino acid sequence of SEQ ID NO: 1018. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1019 and a light chain comprising the amino acid sequence of SEQ ID NO: 1020. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1021 and a light chain comprising the amino acid sequence of SEQ ID NO: 1022. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1023 and a light chain comprising the amino acid sequence of SEQ ID NO: 1024. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1025 and a light chain comprising the amino acid sequence of SEQ ID NO: 1026. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1027 and a light chain comprising the amino acid sequence of SEQ ID NO: 1028. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1029 and a light chain comprising the amino acid sequence of SEQ ID NO: 1030. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1031 and a light chain comprising the amino acid sequence of SEQ ID NO: 1032. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1033 and a light chain comprising the amino acid sequence of SEQ ID NO: 1034. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1035 and a light chain comprising the amino acid sequence of SEQ ID NO: 1036. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1037 and a light chain comprising the amino acid sequence of SEQ ID NO: 1038.In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1039 and a light chain comprising the amino acid sequence of SEQ ID NO: 1040. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1041 and a light chain comprising the amino acid sequence of SEQ ID NO: 1042. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1043 and a light chain comprising the amino acid sequence of SEQ ID NO: 1044. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1045 and a light chain comprising the amino acid sequence of SEQ ID NO: 1046. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1047 and a light chain comprising the amino acid sequence of SEQ ID NO: 1048. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1049 and a light chain comprising the amino acid sequence of SEQ ID NO: 1050. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1051 and a light chain comprising the amino acid sequence of SEQ ID NO: 1052. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1053 and a light chain comprising the amino acid sequence of SEQ ID NO: 1054. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1055 and a light chain comprising the amino acid sequence of SEQ ID NO: 1056. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1057 and a light chain comprising the amino acid sequence of SEQ ID NO: 1058. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1059 and a light chain comprising the amino acid sequence of SEQ ID NO: 1060. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1061 and a light chain comprising the amino acid sequence of SEQ ID NO: 1062.In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1063 and a light chain comprising the amino acid sequence of SEQ ID NO: 1064. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1065 and a light chain comprising the amino acid sequence of SEQ ID NO: 1066. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1067 and a light chain comprising the amino acid sequence of SEQ ID NO: 1068. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1069 and a light chain comprising the amino acid sequence of SEQ ID NO: 1070. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1071 and a light chain comprising the amino acid sequence of SEQ ID NO: 1072. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1073 and a light chain comprising the amino acid sequence of SEQ ID NO: 1074. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1075 and a light chain comprising the amino acid sequence of SEQ ID NO: 1076. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1077 and a light chain comprising the amino acid sequence of SEQ ID NO: 1078. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1079 and a light chain comprising the amino acid sequence of SEQ ID NO: 1080. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1081 and a light chain comprising the amino acid sequence of SEQ ID NO: 1082. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1083 and a light chain comprising the amino acid sequence of SEQ ID NO: 1084. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1085 and a light chain comprising the amino acid sequence of SEQ ID NO: 1086.In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1087 and a light chain comprising the amino acid sequence of SEQ ID NO: 1088. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1089 and a light chain comprising the amino acid sequence of SEQ ID NO: 1090. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1091 and a light chain comprising the amino acid sequence of SEQ ID NO: 1092. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1093 and a light chain comprising the amino acid sequence of SEQ ID NO: 1094. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1095 and a light chain comprising the amino acid sequence of SEQ ID NO: 1096. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1097 and a light chain comprising the amino acid sequence of SEQ ID NO: 1098. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1099 and a light chain comprising the amino acid sequence of SEQ ID NO: 1100. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1101 and a light chain comprising the amino acid sequence of SEQ ID NO: 1102. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1103 and a light chain comprising the amino acid sequence of SEQ ID NO: 1104. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1105 and a light chain comprising the amino acid sequence of SEQ ID NO: 1106. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1107 and a light chain comprising the amino acid sequence of SEQ ID NO: 1108.

[0703] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-ANXA1-anti-CD47]

[0704] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody that binds to ANXA1 and a second domain (third component) including a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2) and a light chain constant region (CL) of an antibody that binds to CD47 are serially connected through a first linker (second component), and an Fc region or a variant thereof (fourth component) is connected through a third linker.

[0705] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, wherein a sixth structure comprises a heavy chain variable region (VH1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to CD47, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprises a light chain variable region (VL1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to CD47, a type 1 non-cleavable linker, and a light chain constant region (CL).

[0706] In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1109 and a light chain comprising the amino acid sequence of SEQ ID NO: 1110. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1111 and a light chain comprising the amino acid sequence of SEQ ID NO: 1112. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1113 and a light chain comprising the amino acid sequence of SEQ ID NO: 1114. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1115 and a light chain comprising the amino acid sequence of SEQ ID NO: 1116. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1117 and a light chain comprising the amino acid sequence of SEQ ID NO: 1118. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1119 and a light chain comprising the amino acid sequence of SEQ ID NO: 1120. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1121 and a light chain comprising the amino acid sequence of SEQ ID NO: 1122. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1123 and a light chain comprising the amino acid sequence of SEQ ID NO: 1124. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1125 and a light chain comprising the amino acid sequence of SEQ ID NO: 1126. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1127 and a light chain comprising the amino acid sequence of SEQ ID NO: 1128. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1129 and a light chain comprising the amino acid sequence of SEQ ID NO: 1130. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1131 and a light chain comprising the amino acid sequence of SEQ ID NO: 1132.In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1133 and a light chain comprising the amino acid sequence of SEQ ID NO: 1134. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1135 and a light chain comprising the amino acid sequence of SEQ ID NO: 1136.

[0707] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-ANXA1-anti-EGFR]

[0708] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody that binds to ANXA1 and a second domain (third component) including a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2) and a light chain constant region (CL) of an antibody that binds to EGFR are serially connected through a first linker (second component), and an Fc region or a variant thereof (fourth component) is connected through a third linker.

[0709] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, wherein a sixth structure comprises a heavy chain variable region (VH1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to EGFR, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprises a light chain variable region (VL1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to EGFR, a type 1 non-cleavable linker, and a light chain constant region (CL). In addition, the multi-specific fusion protein may sequentially include a type 1 non-cleavable linker and a peptide motif at the C-terminus of the light chain constant region (CL).

[0710] In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1137 and a light chain comprising the amino acid sequence of SEQ ID NO: 1138. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1139 and a light chain comprising the amino acid sequence of SEQ ID NO: 1140. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1141 and a light chain comprising the amino acid sequence of SEQ ID NO: 1142. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1143 and a light chain comprising the amino acid sequence of SEQ ID NO: 1144. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1145 and a light chain comprising the amino acid sequence of SEQ ID NO: 1146. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1147 and a light chain comprising the amino acid sequence of SEQ ID NO: 1148. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1149 and a light chain comprising the amino acid sequence of SEQ ID NO: 1150. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1151 and a light chain comprising the amino acid sequence of SEQ ID NO: 1152. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1153 and a light chain comprising the amino acid sequence of SEQ ID NO: 1154. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1155 and a light chain comprising the amino acid sequence of SEQ ID NO: 1156. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1157 and a light chain comprising the amino acid sequence of SEQ ID NO: 1158. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1159 and a light chain comprising the amino acid sequence of SEQ ID NO: 1160.In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1161 and a light chain comprising the amino acid sequence of SEQ ID NO: 1162. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1163 and a light chain comprising the amino acid sequence of SEQ ID NO: 1164. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1165 and a light chain comprising the amino acid sequence of SEQ ID NO: 1166. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1167 and a light chain comprising the amino acid sequence of SEQ ID NO: 1168. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1169 and a light chain comprising the amino acid sequence of SEQ ID NO: 1170. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1171 and a light chain comprising the amino acid sequence of SEQ ID NO: 1172. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1173 and a light chain comprising the amino acid sequence of SEQ ID NO: 1174. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1175 and a light chain comprising the amino acid sequence of SEQ ID NO: 1176. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1177 and a light chain comprising the amino acid sequence of SEQ ID NO: 1178. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1179 and a light chain comprising the amino acid sequence of SEQ ID NO: 1180. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1181 and a light chain comprising the amino acid sequence of SEQ ID NO: 1182. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1183 and a light chain comprising the amino acid sequence of SEQ ID NO: 1184.In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1185 and a light chain comprising the amino acid sequence of SEQ ID NO: 1186. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1187 and a light chain comprising the amino acid sequence of SEQ ID NO: 1188. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1159 and a light chain comprising the amino acid sequence of SEQ ID NO: 1189. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1167 and a light chain comprising the amino acid sequence of SEQ ID NO: 1190. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1191 and a light chain comprising the amino acid sequence of SEQ ID NO: 1192. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1193 and a light chain comprising the amino acid sequence of SEQ ID NO: 1194. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1195 and a light chain comprising the amino acid sequence of SEQ ID NO: 1196. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1197 and a light chain comprising the amino acid sequence of SEQ ID NO: 1198. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1199 and a light chain comprising the amino acid sequence of SEQ ID NO: 1200. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1201 and a light chain comprising the amino acid sequence of SEQ ID NO: 1202. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1203 and a light chain comprising the amino acid sequence of SEQ ID NO: 1204. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1205 and a light chain comprising the amino acid sequence of SEQ ID NO: 1206.In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1207 and a light chain comprising the amino acid sequence of SEQ ID NO: 1208.

[0711] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-ANXA1-anti-DLK1]

[0712] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody that binds to ANXA1 and a second domain (third component) including a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2) and a light chain constant region (CL) of an antibody that binds to DLK1 are serially connected through a first linker (second component), and an Fc region or a variant thereof (fourth component) is connected through a third linker.

[0713] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, wherein a sixth structure comprises a heavy chain variable region (VH1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to DLK1, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprises a light chain variable region (VL1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to DLK1, a type 1 non-cleavable linker, and a light chain constant region (CL). In addition, the multi-specific fusion protein may sequentially include a type 1 non-cleavable linker and a peptide motif at the C-terminus of the light chain constant region (CL).

[0714] In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1209 and a light chain comprising the amino acid sequence of SEQ ID NO: 1210. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1211 and a light chain comprising the amino acid sequence of SEQ ID NO: 1212. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1213 and a light chain comprising the amino acid sequence of SEQ ID NO: 1214. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1215 and a light chain comprising the amino acid sequence of SEQ ID NO: 1216. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1217 and a light chain comprising the amino acid sequence of SEQ ID NO: 1218. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1219 and a light chain comprising the amino acid sequence of SEQ ID NO: 1220. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1221 and a light chain comprising the amino acid sequence of SEQ ID NO: 1222. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1223 and a light chain comprising the amino acid sequence of SEQ ID NO: 1224. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1225 and a light chain comprising the amino acid sequence of SEQ ID NO: 1226. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1209 and a light chain comprising the amino acid sequence of SEQ ID NO: 1227. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1211 and a light chain comprising the amino acid sequence of SEQ ID NO: 1228. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1213 and a light chain comprising the amino acid sequence of SEQ ID NO: 1229.In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1215 and a light chain comprising the amino acid sequence of SEQ ID NO: 1230. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1217 and a light chain comprising the amino acid sequence of SEQ ID NO: 1231. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1219 and a light chain comprising the amino acid sequence of SEQ ID NO: 1232. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1221 and a light chain comprising the amino acid sequence of SEQ ID NO: 1233. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1223 and a light chain comprising the amino acid sequence of SEQ ID NO: 1234. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1225 and a light chain comprising the amino acid sequence of SEQ ID NO: 1235. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1236 and a light chain comprising the amino acid sequence of SEQ ID NO: 1237. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1238 and a light chain comprising the amino acid sequence of SEQ ID NO: 1239.

[0715] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-ANXA1-anti-CLDN18.2]

[0716] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody that binds to ANXA1 and a second domain (third component) including a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2) and a light chain constant region (CL) of an antibody that binds to CLDN18.2 are serially connected through a first linker (second component), and an Fc region or a variant thereof (fourth component) is connected through a third linker.

[0717] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, wherein a sixth structure comprises a heavy chain variable region (VH1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to CLDN18.2, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprises a light chain variable region (VL1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to CLDN18.2, a type 1 non-cleavable linker, and a light chain constant region (CL). The Fc region (human IgG1) may be in a form in which the amino acid of CH2 is substituted with L234A / L235A (LALA).

[0718] In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1240 and a light chain comprising the amino acid sequence of SEQ ID NO: 1241. In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1242 and a light chain comprising the amino acid sequence of SEQ ID NO: 1243.

[0719] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-ANXA1-anti-TEM8]

[0720] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region and a light chain variable region of an antibody binding to ANXA1 and a second domain (third component) including a heavy chain variable region, a heavy chain constant region, a light chain variable region, and a light chain constant region of an antibody binding to TEM8 are serially connected through a first linker (second component) and an Fc region or a variant thereof (fourth component) is connected through a third linker. In this case,

[0721] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, wherein a sixth structure comprises a heavy chain variable region (VH1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to TEM8, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprises a light chain variable region (VL1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to TEM8, a type 1 non-cleavable linker, and a light chain constant region (CL). The Fc region (human IgG1) may be in a form in which the amino acid of CH2 is substituted with L234A / L235A (LALA).

[0722] In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1244 and a light chain comprising the amino acid sequence of SEQ ID NO: 1245.

[0723] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-ANXA1-anti-FAPα]

[0724] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody that binds to ANXA1 and a second domain (third component) including a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2) and a light chain constant region (CL) of an antibody that binds to FAPα are serially connected through a first linker (second component), and an Fc region or a variant thereof (fourth component) is connected through a third linker.

[0725] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, wherein a sixth structure comprises a heavy chain variable region (VH1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to FAPα, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprises a light chain variable region (VL1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to FAPα, a type 1 non-cleavable linker, and a light chain constant region (CL). The Fc region (human IgG1) may be in a form in which the amino acid of CH2 is substituted with L234A / L235A (LALA).

[0726] In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1246 and a light chain comprising the amino acid sequence of SEQ ID NO: 1247.

[0727] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-ANXA1-anti-EpCAM]

[0728] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody that binds to ANXA1 and a second domain (third component) including a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2) and a light chain constant region (CL) of an antibody that binds to EpCAM are serially connected through a first linker (second component), and an Fc region or a variant thereof (fourth component) is connected through a third linker.

[0729] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, wherein a sixth structure comprises a heavy chain variable region (VH1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to EpCAM, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprises a light chain variable region (VL1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to EpCAM, a type 1 non-cleavable linker, and a light chain constant region (CL). The Fc region (human IgG1) may be in a form in which the amino acid of CH2 is substituted with L234A / L235A (LALA).

[0730] In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1248 and a light chain comprising the amino acid sequence of SEQ ID NO: 1249.

[0731] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-ANXA1-anti-PD-1]

[0732] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody that binds to ANXA1 and a second domain (third component) including a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2) and a light chain constant region (CL) of an antibody that binds to PD-1 are serially connected through a first linker (second component), and an Fc region or a variant thereof (fourth component) is connected through a third linker.

[0733] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, wherein a sixth structure comprises a heavy chain variable region (VH1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to PD-1, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprises a light chain variable region (VL1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to PD-1, a type 1 non-cleavable linker, and a light chain constant region (CL). The Fc region (human IgG1) may be in a form in which the amino acid of CH2 is substituted with L234A / L235A (LALA).

[0734] In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1250 and a light chain comprising the amino acid sequence of SEQ ID NO: 1251.

[0735] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-ANXA1-anti-PD-L1]

[0736] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody that binds to ANXA1 and a second domain (third component) including a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2) and a light chain constant region (CL) of an antibody that binds to PD-L1 are serially connected through a first linker (second component), and an Fc region or a variant thereof (fourth component) is connected through a third linker.

[0737] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, wherein a sixth structure comprises a heavy chain variable region (VH1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to PD-L1, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprises a light chain variable region (VL1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to PD-L1, a type 1 non-cleavable linker, and a light chain constant region (CL). The Fc region (human IgG1) may be in a form in which the amino acid of CH2 is substituted with L234A / L235A (LALA).

[0738] In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1252 and a light chain comprising the amino acid sequence of SEQ ID NO: 1253.

[0739] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-ANXA1-anti-Aβ protofibrils]

[0740] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody that binds to ANXA1 and a second domain (third component) including a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2) and a light chain constant region (CL) of an antibody that binds to Aβ protofibrils are serially connected through a first linker (second component), and an Fc region or a variant thereof (fourth component) is connected through a third linker.

[0741] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, wherein a sixth structure comprises a heavy chain variable region (VH1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to Aβ protofibrils, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprises a light chain variable region (VL1) of an antibody that binds to ANXA1, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to Aβ protofibrils, a type 1 non-cleavable linker, and a light chain constant region (CL). The Fc region (human IgG1) may be in a form in which the amino acid of CH2 is substituted with L234A / L235A (LALA).

[0742] In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1254 and a light chain comprising the amino acid sequence of SEQ ID NO: 1255.

[0743] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-TEM8-anti-EGFR]

[0744] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody that binds to TEM8 and a second domain (third component) including a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2) and a light chain constant region (CL) of an antibody that binds to EGFR are serially connected through a first linker (second component), and an Fc region or a variant thereof (fourth component) is connected through a third linker.

[0745] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, each of which comprises a sixth structure comprising a heavy chain variable region (VH1) of an antibody that binds to TEM8, a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to EGFR, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprising a light chain variable region (VL1) of an antibody that binds to TEM8, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to EGFR, a type 1 non-cleavable linker, and a light chain constant region (CL). The Fc region (human IgG1) may be in a form in which the amino acid of CH2 is substituted with L234A / L235A (LALA).

[0746] In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1256 and a light chain comprising the amino acid sequence of SEQ ID NO: 1257.

[0747] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-TEM8-anti-ANXA1]

[0748] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody that binds to TEM8 and a second domain (third component) including a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2) and a light chain constant region (CL) of an antibody that binds to ANXA1 are serially connected through a first linker (second component), and an Fc region or a variant thereof (fourth component) is connected through a third linker.

[0749] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, wherein a sixth structure comprises a heavy chain variable region (VH1) of an antibody that binds to TEM8, a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to ANXA1, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprises a light chain variable region (VL1) of an antibody that binds to TEM8, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to ANXA1, a type 1 non-cleavable linker, and a light chain constant region (CL). The Fc region (human IgG1) may be in a form in which the amino acid of CH2 is substituted with L234A / L235A (LALA).

[0750] In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1258 and a light chain comprising the amino acid sequence of SEQ ID NO: 1259.

[0751] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-Albumin-anti-EGFR]

[0752] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody that binds to Albumin and a second domain (third component) including a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2) and a light chain constant region (CL) of an antibody that binds to EGFR are serially connected through a first linker (second component), and an Fc region or a variant thereof (fourth component) is connected through a third linker.

[0753] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, each of which comprises a sixth structure comprising a heavy chain variable region (VH1) of an antibody that binds to Albumin, a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to EGFR, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprising a light chain variable region (VL1) of an antibody that binds to Albumin, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to EGFR, a type 1 non-cleavable linker, and a light chain constant region (CL). The Fc region (human IgG1) may be in a form in which the amino acid of CH2 is substituted with L234A / L235A (LALA).

[0754] In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1260 and a light chain comprising the amino acid sequence of SEQ ID NO: 1261.

[0755] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-KDR (VEGFR2)-anti-EGFR]

[0756] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody that binds to KDR (VEGFR2) and a second domain (third component) including a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2) and a light chain constant region (CL) of an antibody that binds to EGFR are serially connected through a first linker (second component), and an Fc region or a variant thereof (fourth component) is connected through a third linker.

[0757] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, each of which comprises a sixth structure comprising a heavy chain variable region (VH1) of an antibody that binds to KDR (VEGFR2), a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to EGFR, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprising a light chain variable region (VL1) of an antibody that binds to KDR, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to EGFR, a type 1 non-cleavable linker, and a light chain constant region (CL). The Fc region (human IgG1) may be in a form in which the amino acid of CH2 is substituted with L234A / L235A (LALA).

[0758] In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1262 and a light chain comprising the amino acid sequence of SEQ ID NO: 1263.

[0759] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-PSMA-anti-EGFR]

[0760] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody that binds to PSMA and a second domain (third component) including a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2) and a light chain constant region (CL) of an antibody that binds to EGFR are serially connected through a first linker (second component), and an Fc region or a variant thereof (fourth component) is connected through a third linker.

[0761] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, each of which comprises a sixth structure comprising a heavy chain variable region (VH1) of an antibody that binds to PSMA, a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to EGFR, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprising a light chain variable region (VL1) of an antibody that binds to PSMA, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to EGFR, a type 1 non-cleavable linker, and a light chain constant region (CL). The Fc region (human IgG1) may be in a form in which the amino acid of CH2 is substituted with L234A / L235A (LALA).

[0762] In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1264 and a light chain comprising the amino acid sequence of SEQ ID NO: 1265.

[0763] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-ICAM1-anti-EGFR]

[0764] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody that binds to ICAM1 and a second domain (third component) including a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2) and a light chain constant region (CL) of an antibody that binds to EGFR are serially connected through a first linker (second component), and an Fc region or a variant thereof (fourth component) is connected through a third linker.

[0765] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, wherein a sixth structure comprises a heavy chain variable region (VH1) of an antibody that binds to ICAM1, a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to EGFR, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprises a light chain variable region (VL1) of an antibody that binds to ICAM1, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to EGFR, a type 1 non-cleavable linker, and a light chain constant region (CL). The Fc region (human IgG1) may be in a form in which the amino acid of CH2 is substituted with L234A / L235A (LALA).

[0766] In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1266 and a light chain comprising the amino acid sequence of SEQ ID NO: 1267.

[0767] -Multi-specific fusion protein in the form of DVD-Ig containing ReleaDome [anti-FAPα-anti-EGFR]

[0768] The above multi-specific fusion protein may be a ReleaDome in which a first domain (first component) including a heavy chain variable region (VH1) and a light chain variable region (VL1) of an antibody that binds to FAPα and a second domain (third component) including a heavy chain variable region (VH2), a heavy chain constant region (CH1), a light chain variable region (VL2) and a light chain constant region (CL) of an antibody that binds to EGFR are serially connected through a first linker (second component), and an Fc region or a variant thereof (fourth component) is connected through a third linker.

[0769] Specifically, the multi-specific fusion protein dimer may be a combination of two multi-specific fusion proteins, wherein a sixth structure comprises a heavy chain variable region (VH1) of an antibody that binds to FAPα, a type 1 cleavable linker, a heavy chain variable region (VH2) of an antibody that binds to EGFR, a type 1 non-cleavable linker, a heavy chain constant region (CH1), a third linker, and an Fc region or a variant thereof, and a second structure comprises a light chain variable region (VL1) of an antibody that binds to FAPα, a type 1 cleavable linker, a light chain variable region (VL2) of an antibody that binds to EGFR, a type 1 non-cleavable linker, and a light chain constant region (CL). The Fc region (human IgG1) may be in a form in which the amino acid of CH2 is substituted with L234A / L235A (LALA).

[0770] In one specific example, it may include a heavy chain comprising the amino acid sequence of SEQ ID NO: 1268 and a light chain comprising the amino acid sequence of SEQ ID N...

Claims

1. A fusion protein comprising a first domain comprising a site that specifically binds to a first target; and a second domain comprising a site that specifically binds to an immune modulator or a second target; A fusion protein wherein the first target is a protein overexpressed in vascular cells or basal cells at the site of the disease, and the second target is a protein overexpressed or secreted on the surface of cancer cells or immune cells.

2. In paragraph 1, A fusion protein characterized in that the first domain shields the second domain at a non-diseased site and / or is cleaved at a diseased site to induce the activity of the second domain.

3. In paragraph 1, A fusion protein, wherein the first domain comprises a site that binds to any one protein selected from the group consisting of Annexin A1, Albumin, App2, B7H3, Cav-1, CD36, EphA2, EphA3, EphB1, FAPα (Fibroblast activation protein alpha), ICAM1, Integrins, KDR (VEGF2), TEM8, TSP-1, ED-B, Endothelin-1, Endoglin (CD105), VEGFR1 (FLT1), PDGFR, PSMA, and VCAM-1.

4. In paragraph 1, A fusion protein, wherein the first domain is any one selected from the group consisting of an antibody or a fragment thereof, a ligand or a fragment thereof, a receptor or a fragment thereof, a peptide, and an aptamer.

5. In paragraph 4, A fusion protein, wherein the first domain is an antibody or a fragment thereof.

6. In paragraph 5, The above first domain is HCDR1 comprising any one amino acid sequence selected from the group consisting of SEQ ID NOs: 277, 284, 291, 298, 304, 310, 320, 326, 332, 336, 338, 344, 355, 370, 382, 392, 396, 398, 405, 411, 417, 420, 427, 432, 436, 441, 444, 449, 456, 460, 465, 470, 475, 478, and 489; An HCDR2 comprising any one amino acid sequence selected from the group consisting of SEQ ID NOs: 279, 286, 293, 300, 305, 322, 328, 334, 340, 346, 351, 357, 365, 372, 378, 386, 394, 400, 406, 413, 422, 429, 434, 442, 446, 451, 458, 462, 467, 472, 480, 486, 491, 494, 495, 496, 497, and 498, and A heavy chain variable region comprising an HCDR3 comprising any one amino acid sequence selected from the group consisting of SEQ ID NOs: 281, 288, 295, 302, 307, 312, 317, 324, 330, 342, 348, 353, 359, 361, 362, 374, 380, 388, 402, 408, 415, 424, 453, and 492; and LCDR1 comprising any one amino acid sequence selected from the group consisting of sequence numbers 500, 512, 525, 531, 538, 551, 557, 564, 570, 575, 580, 592, 595, 599, 603, 608, 616, 621, 625, 631, 636, 645, 651, 654, 659, 667, 670, 675, 678, 684, 690, 698, 704, 708, 711 and 714; LCDR2 comprising any one amino acid sequence selected from the group consisting of sequence numbers 502, 508, 514, 523, 527, 533, 540, 547, 553, 559, 566, 582, 587, 609, 617, 632, 639, 647, 663, 680, 685 and 716, and A light chain variable region comprising LCDR3 comprising any one amino acid sequence selected from the group consisting of SEQ ID NOs: 504, 510, 516, 529, 535, 542, 544, 549, 555, 561, 568, 572, 578, 584, 586, 588, 590, 597, 602, 606, 611, 613, 619, 624, 627, 630, 634, 641, 643, 649, 653, 662, 665, 669, 673, 674, 682, 687, 689, 693, 696, 697, 718 and 719 A fusion protein comprising:

7. In paragraph 4, The above first domain is A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 277, HCDR2 comprising the amino acid sequence of SEQ ID NO: 279, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 281, and a light chain variable region (A1) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 500, LCDR2 comprising the amino acid sequence of SEQ ID NO: 502, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 504; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 284, HCDR2 comprising the amino acid sequence of SEQ ID NO: 286, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 288, and a light chain variable region (A2) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 500, LCDR2 comprising the amino acid sequence of SEQ ID NO: 508, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 510; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A3) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 516; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 298, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 300, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 302, and a light chain variable region (A4) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 500, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 502, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 504; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 304, HCDR2 comprising the amino acid sequence of SEQ ID NO: 305, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 307, and a light chain variable region (A5) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 500, LCDR2 comprising the amino acid sequence of SEQ ID NO: 523, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 510; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 310, HCDR2 comprising the amino acid sequence of SEQ ID NO: 286, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 312, and a light chain variable region (A6) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 525, LCDR2 comprising the amino acid sequence of SEQ ID NO: 527, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 529; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 317, and a light chain variable region (A7) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 531, LCDR2 comprising the amino acid sequence of SEQ ID NO: 533, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 535; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 320, HCDR2 comprising the amino acid sequence of SEQ ID NO: 322, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 324, and a light chain variable region (A8) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 538, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 542; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 326, HCDR2 comprising the amino acid sequence of SEQ ID NO: 328, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 330, and a light chain variable region (A9) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 500, LCDR2 comprising the amino acid sequence of SEQ ID NO: 502, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 544; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 332, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 334, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A10) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 547, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 549; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 336, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A11) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 551, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 553, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 555; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 338, HCDR2 comprising the amino acid sequence of SEQ ID NO: 340, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 342, and a light chain variable region (A12) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 557, LCDR2 comprising the amino acid sequence of SEQ ID NO: 559, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 561; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 344, HCDR2 comprising the amino acid sequence of SEQ ID NO: 346, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 348, and a light chain variable region (A13) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 564, LCDR2 comprising the amino acid sequence of SEQ ID NO: 566, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 568; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 351, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 353, and a light chain variable region (A14) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 570, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 555; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 355, HCDR2 comprising the amino acid sequence of SEQ ID NO: 357, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region (A15) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 572; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 361, and a light chain variable region (A16) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 575, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 578; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 362, and a light chain variable region (A17) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 580, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 582, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 584; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 365, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A18) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 586; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A19) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 587, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 588; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 370, HCDR2 comprising the amino acid sequence of SEQ ID NO: 372, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 374, and a light chain variable region (A20) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 564, LCDR2 comprising the amino acid sequence of SEQ ID NO: 582, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 590; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A21) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 592, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 572; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 355, HCDR2 comprising the amino acid sequence of SEQ ID NO: 378, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 380, and a light chain variable region (A22) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 595, LCDR2 comprising the amino acid sequence of SEQ ID NO: 582, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 597; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 382, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A23) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 572; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 286, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region (A24) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 599, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 582, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 602; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 386, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 388, and a light chain variable region (A25) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 603, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 606; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region (A26) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 608, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 609, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 611; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A27) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 613; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 392, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 394, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A28) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 616, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 617, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 619; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A29) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 621, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 602; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A30) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 608, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 624; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 396, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A31) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 625, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 627; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 398, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 400, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 402, and a light chain variable region (A32) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 625, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 630; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 405, HCDR2 comprising the amino acid sequence of SEQ ID NO: 406, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 408, and a light chain variable region (A33) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 631, LCDR2 comprising the amino acid sequence of SEQ ID NO: 632, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 634; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 411, HCDR2 comprising the amino acid sequence of SEQ ID NO: 413, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 415, and a light chain variable region (A34) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 636, LCDR2 comprising the amino acid sequence of SEQ ID NO: 639, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 641; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, HCDR2 comprising the amino acid sequence of SEQ ID NO: 351, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A35) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 603, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 643; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 420, HCDR2 comprising the amino acid sequence of SEQ ID NO: 422, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 424, and a light chain variable region (A36) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 645, LCDR2 comprising the amino acid sequence of SEQ ID NO: 647, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 649; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 320, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A37) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 651, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 653; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 427, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 429, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A38) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 654, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 602; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 432, HCDR2 comprising the amino acid sequence of SEQ ID NO: 434, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region (A39) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 624; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 436, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A40) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 659, LCDR2 comprising the amino acid sequence of SEQ ID NO: 582, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 662; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, HCDR2 comprising the amino acid sequence of SEQ ID NO: 351, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A41) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 531, LCDR2 comprising the amino acid sequence of SEQ ID NO: 663, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 665; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A42) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 667, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 669; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 320, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A43) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 670, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 673; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A44) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 674; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 441, HCDR2 comprising the amino acid sequence of SEQ ID NO: 442, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A45) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 675, LCDR2 comprising the amino acid sequence of SEQ ID NO: 553, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 643; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 444, HCDR2 comprising the amino acid sequence of SEQ ID NO: 446, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region (A46) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 678, LCDR2 comprising the amino acid sequence of SEQ ID NO: 680, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 682; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 449, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 451, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 453, and a light chain variable region (A47) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 684, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 685, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 687; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 456, HCDR2 comprising the amino acid sequence of SEQ ID NO: 458, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region (A48) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 689; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 460, HCDR2 comprising the amino acid sequence of SEQ ID NO: 462, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A49) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 690, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 693; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 465, HCDR2 comprising the amino acid sequence of SEQ ID NO: 467, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A50) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 696; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 470, HCDR2 comprising the amino acid sequence of SEQ ID NO: 472, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region (A51) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 609, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 697; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 475, HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A52) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 698, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 627; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 478, HCDR2 comprising the amino acid sequence of SEQ ID NO: 480, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region (A53) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 514, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 696; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 293, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A54) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 704, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 602; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 351, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A55 / A69 / A77 / A77-1 / A79 / A82) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 708, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 602; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 351, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 359, and a light chain variable region (A56) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 711, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 602; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 291, HCDR2 comprising the amino acid sequence of SEQ ID NO: 486, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A57) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 678, LCDR2 comprising the amino acid sequence of SEQ ID NO: 680, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 682; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 489, HCDR2 comprising the amino acid sequence of SEQ ID NO: 491, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 492, and a light chain variable region (A58) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 714, LCDR2 comprising the amino acid sequence of SEQ ID NO: 716, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 718; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 351, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A59) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 609, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 719; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, HCDR2 comprising the amino acid sequence of SEQ ID NO: 351, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A60) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 512, LCDR2 comprising the amino acid sequence of SEQ ID NO: 609, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 719; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, HCDR2 comprising the amino acid sequence of SEQ ID NO: 494, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A61 / A62) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 708, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 602; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 495, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A63) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 708, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 602; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, HCDR2 comprising the amino acid sequence of SEQ ID NO: 496, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A64 / A65) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 708, LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 602; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 497, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A66) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 708, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 602; or A fusion protein comprising a heavy chain variable region comprising an HCDR1 comprising an amino acid sequence of SEQ ID NO: 417, an HCDR2 comprising an amino acid sequence of SEQ ID NO: 498, and an HCDR3 comprising an amino acid sequence of SEQ ID NO: 295, and a light chain variable region (A67 / A68) comprising an LCDR1 comprising an amino acid sequence of SEQ ID NO: 708, an LCDR2 comprising an amino acid sequence of SEQ ID NO: 540, and an LCDR3 comprising an amino acid sequence of SEQ ID NO:

602.

8. In paragraph 7, The above first domain is A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1 and a light chain variable region (A1) comprising the amino acid sequence of SEQ ID NO: 2, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 3 and a light chain variable region (A2) comprising the amino acid sequence of SEQ ID NO: 4, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 5 and a light chain variable region (A3) comprising the amino acid sequence of SEQ ID NO: 6, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7 and a light chain variable region (A4) comprising the amino acid sequence of SEQ ID NO: 8; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 9 and a light chain variable region (A5) comprising the amino acid sequence of SEQ ID NO: 10; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 11 and a light chain variable region (A6) comprising the amino acid sequence of SEQ ID NO: 12; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 13 and a light chain variable region (A7) comprising the amino acid sequence of SEQ ID NO: 14, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 15 and a light chain variable region (A8) comprising the amino acid sequence of SEQ ID NO: 16, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 17 and a light chain variable region (A9) comprising the amino acid sequence of SEQ ID NO: 18; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and a light chain variable region (A10) comprising the amino acid sequence of SEQ ID NO: 20; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 21 and a light chain variable region (A11) comprising the amino acid sequence of SEQ ID NO: 22, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 23 and a light chain variable region (A12) comprising the amino acid sequence of SEQ ID NO: 24, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 25 and a light chain variable region (A13) comprising the amino acid sequence of SEQ ID NO: 26, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 27 and a light chain variable region (A14) comprising the amino acid sequence of SEQ ID NO: 28; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 29 and a light chain variable region (A15) comprising the amino acid sequence of SEQ ID NO: 30; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 31 and a light chain variable region (A16) comprising the amino acid sequence of SEQ ID NO: 32, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 33 and a light chain variable region (A17) comprising the amino acid sequence of SEQ ID NO: 34, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 35 and a light chain variable region (A18) comprising the amino acid sequence of SEQ ID NO: 36, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 37 and a light chain variable region (A19) comprising the amino acid sequence of SEQ ID NO: 38; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 39 and a light chain variable region (A20) comprising the amino acid sequence of SEQ ID NO: 40; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 41 and a light chain variable region (A21) comprising the amino acid sequence of SEQ ID NO: 42, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 43 and a light chain variable region (A22) comprising the amino acid sequence of SEQ ID NO: 44, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 45 and a light chain variable region (A23) comprising the amino acid sequence of SEQ ID NO: 46, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 47 and a light chain variable region (A24) comprising the amino acid sequence of SEQ ID NO: 48; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 49 and a light chain variable region (A25) comprising the amino acid sequence of SEQ ID NO: 50; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 51 and a light chain variable region (A26) comprising the amino acid sequence of SEQ ID NO: 52, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 53 and a light chain variable region (A27) comprising the amino acid sequence of SEQ ID NO: 54, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 55 and a light chain variable region (A28) comprising the amino acid sequence of SEQ ID NO: 56; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 57 and a light chain variable region (A29) comprising the amino acid sequence of SEQ ID NO: 58; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 59 and a light chain variable region (A30) comprising the amino acid sequence of SEQ ID NO: 60, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 61 and a light chain variable region (A31) comprising the amino acid sequence of SEQ ID NO: 62, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 63 and a light chain variable region (A32) comprising the amino acid sequence of SEQ ID NO: 64, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 65 and a light chain variable region (A33) comprising the amino acid sequence of SEQ ID NO: 66, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 67 and a light chain variable region (A34) comprising the amino acid sequence of SEQ ID NO: 68; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 69 and a light chain variable region (A35) comprising the amino acid sequence of SEQ ID NO: 70; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 71 and a light chain variable region (A36) comprising the amino acid sequence of SEQ ID NO: 72, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 73 and a light chain variable region (A37) comprising the amino acid sequence of SEQ ID NO: 74, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 75 and a light chain variable region (A38) comprising the amino acid sequence of SEQ ID NO: 76, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 77 and a light chain variable region (A39) comprising the amino acid sequence of SEQ ID NO: 78; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 79 and a light chain variable region (A40) comprising the amino acid sequence of SEQ ID NO: 80; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 81 and a light chain variable region (A41) comprising the amino acid sequence of SEQ ID NO: 82, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 83 and a light chain variable region (A42) comprising the amino acid sequence of SEQ ID NO: 84, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 85 and a light chain variable region (A43) comprising the amino acid sequence of SEQ ID NO: 86, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 87 and a light chain variable region (A44) comprising the amino acid sequence of SEQ ID NO: 88; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 89 and a light chain variable region (A45) comprising the amino acid sequence of SEQ ID NO: 90; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 91 and a light chain variable region (A46) comprising the amino acid sequence of SEQ ID NO: 92, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 93 and a light chain variable region (A47) comprising the amino acid sequence of SEQ ID NO: 94, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 95 and a light chain variable region (A48) comprising the amino acid sequence of SEQ ID NO: 96, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 97 and a light chain variable region (A49) comprising the amino acid sequence of SEQ ID NO: 98; A heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 99 and a light chain variable region (A50) comprising an amino acid sequence of SEQ ID NO: 100; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 101 and a light chain variable region (A51) comprising the amino acid sequence of SEQ ID NO: 102, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 103 and a light chain variable region (A52) comprising the amino acid sequence of SEQ ID NO: 104, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 105 and a light chain variable region (A53) comprising the amino acid sequence of SEQ ID NO: 106, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 107 and a light chain variable region (A54) comprising the amino acid sequence of SEQ ID NO: 108, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 109 and a light chain variable region (A55 / A82) comprising the amino acid sequence of SEQ ID NO: 110; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 111 and a light chain variable region (A56) comprising the amino acid sequence of SEQ ID NO: 112, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 113 and a light chain variable region (A57) comprising the amino acid sequence of SEQ ID NO: 114, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 115 and a light chain variable region (A58) comprising the amino acid sequence of SEQ ID NO: 116, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 109 and a light chain variable region (A59) comprising the amino acid sequence of SEQ ID NO: 117; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 118 and a light chain variable region (A60) comprising the amino acid sequence of SEQ ID NO: 117; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 119 and a light chain variable region (A61) comprising the amino acid sequence of SEQ ID NO: 110, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 120 and a light chain variable region (A62) comprising the amino acid sequence of SEQ ID NO: 110; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 121 and a light chain variable region (A63) comprising the amino acid sequence of SEQ ID NO: 110, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 122 and a light chain variable region (A64) comprising the amino acid sequence of SEQ ID NO: 110; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 123 and a light chain variable region (A65) comprising the amino acid sequence of SEQ ID NO: 110; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 124 and a light chain variable region (A66) comprising the amino acid sequence of SEQ ID NO: 110; A heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 125 and a light chain variable region (A67) comprising an amino acid sequence of SEQ ID NO: 110, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 126 and a light chain variable region (A68) comprising the amino acid sequence of SEQ ID NO: 110; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 118 and a light chain variable region (A69 / A77 / A77-1) comprising the amino acid sequence of SEQ ID NO: 110, or A fusion protein comprising a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 143 and a light chain variable region (A79) comprising an amino acid sequence of SEQ ID NO:

144.

9. In paragraph 5, A fusion protein comprising a heavy chain variable region (A78) comprising an HCDR1 comprising an amino acid sequence of SEQ ID NO: 291, an HCDR2 comprising an amino acid sequence of SEQ ID NO: 293, and an HCDR3 comprising an amino acid sequence of SEQ ID NO:

295.

10. In paragraph 9, A fusion protein wherein the heavy chain variable region comprises the amino acid sequence (A78) of sequence number 142.

11. In paragraph 5, The above first domain is A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 721, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 723, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 725, and a light chain variable region (A70 / A81) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 763, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 765, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 767; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 727, HCDR2 comprising the amino acid sequence of SEQ ID NO: 729, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 731, and a light chain variable region (A71) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 770, LCDR2 comprising the amino acid sequence of SEQ ID NO: 772, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 774; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 733, HCDR2 comprising the amino acid sequence of SEQ ID NO: 735, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 736, and a light chain variable region (A72) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 775, LCDR2 comprising the amino acid sequence of SEQ ID NO: 566, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 778; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 738, HCDR2 comprising the amino acid sequence of SEQ ID NO: 740, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 742, and a light chain variable region (A73) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 781, LCDR2 comprising the amino acid sequence of SEQ ID NO: 559, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 784; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 745, HCDR2 comprising the amino acid sequence of SEQ ID NO: 747, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 749, and a light chain variable region (A74) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 786, LCDR2 comprising the amino acid sequence of SEQ ID NO: 788, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 790; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 751, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 753, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 755, and a light chain variable region (A75) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 792, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 794, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 796; or A fusion protein comprising a heavy chain variable region comprising an HCDR1 comprising an amino acid sequence of SEQ ID NO: 757, an HCDR2 comprising an amino acid sequence of SEQ ID NO: 759, and an HCDR3 comprising an amino acid sequence of SEQ ID NO: 761, and a light chain variable region comprising an LCDR1 comprising an amino acid sequence of SEQ ID NO: 580, an LCDR2 comprising an amino acid sequence of SEQ ID NO: 566, and an LCDR3 comprising an amino acid sequence of SEQ ID NO: 799 (A76).

12. In paragraph 11, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 127 and a light chain variable region (A70 / A81) comprising the amino acid sequence of SEQ ID NO: 128; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 129 and a light chain variable region (A71) comprising the amino acid sequence of SEQ ID NO: 130, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 131 and a light chain variable region (A72) comprising the amino acid sequence of SEQ ID NO: 132, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 133 and a light chain variable region (A73) comprising the amino acid sequence of SEQ ID NO: 134, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 135 and a light chain variable region (A74) comprising the amino acid sequence of SEQ ID NO: 136, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 137 and a light chain variable region (A75) comprising the amino acid sequence of SEQ ID NO: 138, or A fusion protein comprising a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 139 and a light chain variable region (A76) comprising an amino acid sequence of SEQ ID NO:

140.

13. In paragraph 4, A fusion protein, the first domain of which is a peptide.

14. In paragraph 13, A fusion protein, wherein the first domain comprises the amino acid sequence of sequence number 145 (A80 / A80-1).

15. In paragraph 1, A fusion protein wherein the above immune modulator is a cytokine or chemokine or a fragment thereof.

16. In paragraph 1, 제2 도메인은 4-1-BB, 5T4, ACE, ADAM9, ADRB3, AKAP4, ALK, ANXA1, ANXA2, ANXA5, ASCT2, AXL, B7H3, B7H4, BCL2, BCMA, CA125, CA6, CA9, CCL11, CCR5, CEN106, CD2, CD4, CD5, CD15, CD16, CD19, CD20, CD22, CD23, CD24, CD25, CD27, CD30, CD33, CD37, CD38, CD40, CD41, CD44, CD44v6, CD46, CD48, CD51, CD52, CD54, CD56, CD62, CD70, CD71, CD72, CD73, CD74, CD79, CD80, CD90, CD97, CD99, CD105, CD117, CD123, CD133, CD135, CD138, CD142, CD163, CD171, CD205, CD242, CD319, CD352, CDCP1, CEA, CLDN3, CLDN6, CLDN9, CLDN18.2, CXCR2, CXCR4, CXCR5, CFC1B, CEACAM5, CEACAM6, c-Met, c-Kit, CTLA4, DLL3, DLK-1, DR5, DPEP3, Dectin-2, ED-B, EGFR, Endothelin-1, Endoglin, ENPP3, EpCAM, EphA2, ETA, FAPα, FGFR1, FGFR2, FGFR3, FRα, GD2, GD3, GloboH, GPC3, GPR20, GPRC5D, HER2, HER3, ICAM1, ICAM2, ICOS-L, IGF-1R, KAAG-1, L1CAM, LCK, LGR5, LIV-1, LRRC15, LY6E, MAGE, Mesothelin, MUC1, MUC16, MMP-14, NCAM1, A fusion protein comprising a binding site selected from the group consisting of Nectin-4, Notch3, NaPi2b, PDGFR, PD-L1, PD-L2, PSCA, PSMA, PTK7, RAGE-1, RANKL, RNF43, RON, ROR1, ROR2, SAP-1, SLAMF7, SLC44A4, Survivin, SAIL, STn, SSEA-4, SEZ6, SLC44A4, SLTRK6, TEM1, TEM8, TM4SF1, TNFLYPD3, TF, TIE2, TIM-1, TRAC, TROP-2, VEGFR1(FLT1), VEGFR2(KDR), VCAM-1, VSIG4, VISTA, and XAGE-1.

17. In paragraph 1, A fusion protein, wherein the second domain is any one selected from the group consisting of an antibody or a fragment thereof, a ligand or a fragment thereof, and a receptor or a fragment thereof.

18. In paragraph 17, A fusion protein, wherein the second domain is an antibody or a fragment thereof.

19. In paragraph 18, The second domain is A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 802, HCDR2 comprising the amino acid sequence of SEQ ID NO: 804, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 806, and a light chain variable region (C1 / C2) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 913, LCDR2 comprising the amino acid sequence of SEQ ID NO: 680, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 915; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 809, HCDR2 comprising the amino acid sequence of SEQ ID NO: 811, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 813, and a light chain variable region (C3 / C4) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 918, LCDR2 comprising the amino acid sequence of SEQ ID NO: 920, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 922; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 815, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 817, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 819, and a light chain variable region (C5 / C6) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 926, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 920, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 929; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 821, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 823, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 825, and a light chain variable region (C7 / C8) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 932, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 934, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 936; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 829, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 832, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 834, and a light chain variable region (C9 / C10) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 939, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 941, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 943; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 838, HCDR2 comprising the amino acid sequence of SEQ ID NO: 840, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 842, and a light chain variable region (C11 / C12 / C30) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 903, LCDR2 comprising the amino acid sequence of SEQ ID NO: 566, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 906; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 844, HCDR2 comprising the amino acid sequence of SEQ ID NO: 846, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 848, and a light chain variable region (C13 / C14) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 580, LCDR2 comprising the amino acid sequence of SEQ ID NO: 566, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 937; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 850, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 852, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 854, and a light chain variable region (C15) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 948, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 949, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 951; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 855, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 857, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 858, and a light chain variable region (C16) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 952, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 954; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 757, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 759, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 761, and a light chain variable region (C17 / C18) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 580, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 566, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 799; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 417, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 351, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 295, and a light chain variable region (C19) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 708, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 540, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 602; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 861, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 863, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 865, and a light chain variable region (C20) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 957, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 958, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 960; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 866, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 868, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 870, and a light chain variable region (C21 / C29) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 962, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 964, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 966; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 872, HCDR2 comprising the amino acid sequence of SEQ ID NO: 874, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 876, and a light chain variable region (C22) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 968, LCDR2 comprising the amino acid sequence of SEQ ID NO: 559, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 971; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 878, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 880, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 882, and a light chain variable region (C23) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 580, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 566, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 973; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 721, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 723, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 725, and a light chain variable region (C24) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 763, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 765, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 767; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 751, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 753, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 755, and a light chain variable region (C25) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 792, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 794, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 796; A heavy chain variable region comprising HCDR1 comprising the amino acid sequence of SEQ ID NO: 884, HCDR2 comprising the amino acid sequence of SEQ ID NO: 886, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 888, and a light chain variable region (C26) comprising LCDR1 comprising the amino acid sequence of SEQ ID NO: 975, LCDR2 comprising the amino acid sequence of SEQ ID NO: 977, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 979; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 890, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 892, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 894, and a light chain variable region (C27) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 580, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 566, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 981; A heavy chain variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 895, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 898, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 900, and a light chain variable region (C28 / C31) comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 982, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 680, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 984; or A fusion protein comprising a heavy chain variable region comprising an HCDR1 comprising an amino acid sequence of SEQ ID NO: 908, an HCDR2 comprising an amino acid sequence of SEQ ID NO: 910, and an HCDR3 comprising an amino acid sequence of SEQ ID NO: 912, and a light chain variable region (C33) comprising an LCDR1 comprising an amino acid sequence of SEQ ID NO: 986, an LCDR2 comprising an amino acid sequence of SEQ ID NO: 920, and an LCDR3 comprising an amino acid sequence of SEQ ID NO:

989.

20. In paragraph 19, The above second domain is A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 184 and a light chain variable region (C1) comprising the amino acid sequence of SEQ ID NO: 185, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 186 and a light chain variable region (C2) comprising the amino acid sequence of SEQ ID NO: 187; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 188 and a light chain variable region (C3) comprising the amino acid sequence of SEQ ID NO: 189; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 190 and a light chain variable region (C4) comprising the amino acid sequence of SEQ ID NO: 191; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 192 and a light chain variable region (C5) comprising the amino acid sequence of SEQ ID NO: 193; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 194 and a light chain variable region (C6) comprising the amino acid sequence of SEQ ID NO: 195, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 196 and a light chain variable region (C7) comprising the amino acid sequence of SEQ ID NO: 197, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 198 and a light chain variable region (C8) comprising the amino acid sequence of SEQ ID NO: 199, A heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 200 and a light chain variable region (C9) comprising an amino acid sequence of SEQ ID NO: 201; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 202 and a light chain variable region (C10) comprising the amino acid sequence of SEQ ID NO: 203; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 204 and a light chain variable region (C11 / C30) comprising the amino acid sequence of SEQ ID NO: 205, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 206 and a light chain variable region (C12) comprising the amino acid sequence of SEQ ID NO: 207, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 208 and a light chain variable region (C13) comprising the amino acid sequence of SEQ ID NO: 209, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 210 and a light chain variable region (C14) comprising the amino acid sequence of SEQ ID NO: 211; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 212 and a light chain variable region (C15) comprising the amino acid sequence of SEQ ID NO: 213; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 214 and a light chain variable region (C16) comprising the amino acid sequence of SEQ ID NO: 215, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 139 and a light chain variable region (C17) comprising the amino acid sequence of SEQ ID NO: 140, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 216 and a light chain variable region (C18) comprising the amino acid sequence of SEQ ID NO: 217, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 109 and a light chain variable region (C19) comprising the amino acid sequence of SEQ ID NO: 110; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 218 and a light chain variable region (C20) comprising the amino acid sequence of SEQ ID NO: 219; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 220 and a light chain variable region (C21) comprising the amino acid sequence of SEQ ID NO: 221, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 222 and a light chain variable region (C22) comprising the amino acid sequence of SEQ ID NO: 223, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 224 and a light chain variable region (C23) comprising the amino acid sequence of SEQ ID NO: 225, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 127 and a light chain variable region (C24) comprising the amino acid sequence of SEQ ID NO: 128, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 137 and a light chain variable region (C25) comprising the amino acid sequence of SEQ ID NO: 138, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 226 and a light chain variable region (C26) comprising the amino acid sequence of SEQ ID NO: 227, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 228 and a light chain variable region (C27) comprising the amino acid sequence of SEQ ID NO: 229, A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 230 and a light chain variable region (C28 / C32) comprising the amino acid sequence of SEQ ID NO: 231; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 232 and a light chain variable region (C29) comprising the amino acid sequence of SEQ ID NO: 233; A heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 237 and a light chain variable region (C31) comprising the amino acid sequence of SEQ ID NO: 236, or A fusion protein comprising a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 239 and a light chain variable region (C33) comprising an amino acid sequence of SEQ ID NO:

240.

21. In paragraph 1, A fusion protein wherein the first domain and the second domain are connected by a linker.

22. In paragraph 21, A fusion protein, wherein the linker is a cleavable linker or a non-cleavable linker.

23. In paragraph 21, A fusion protein, wherein the truncated linker comprises any one selected from the group consisting of SEQ ID NO: 159 to SEQ ID NO: 176 and combinations thereof.

24. In paragraph 1, The above fusion protein comprises the following structural formulas (I) and (II): N'-T1'-((L1)a-CR1)b-(L2')-I1'-((L3)c-CR3)dC' (I); N'-T1''-((L1)a-CR2)b-(L2'')-I1''-((L3)c-CR4)dC' (II) At this time, in the structural formulas (I) and (II), The above N' is the N-terminus of the fusion protein, The above C' is the C-terminus of the fusion protein, The above T1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the first target, The above T1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the first target, The above T1' and T1'' combine to form a binding site (T1) that specifically binds to the first target, The above I1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target, The above I1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target, The above I1' and I1'' combine to form a binding site (I1) that specifically binds to the second target, The above CR1, CR2, CR3 and CR4 are each independently CH1 or CL, At this time, CR1 and CR2 are not equal, CR3 and CR4 are not equal, The above CH1 is a heavy chain constant region, The above CL is a light chain constant region, The above L1, L2', L2'' and L3 are each independently a peptide linker, wherein L2' and L2'' are truncated or uncleaved, The above a, b, c and d are each independently 0 or 1.

25. In paragraph 1, The above fusion protein comprises the following structural formulas (IV) and (V): N'-T2-(L7)-I2'-((L8)f-CR1)gC' (IV); N'-I2''-((L8)f-CR2)hC' (V) At this time, in the structural formulas (IV) and (V), The above N' is the N-terminus of the fusion protein, The above C' is the C-terminus of the fusion protein, The above T2 is a peptide or antibody fragment that specifically binds to the first target, The above I2' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target, The above I2'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target, The above I2' and I2'' combine to form a binding site (I2) that specifically binds to the second target, The above CR1 and CR2 are each independently CH1 or CL, At this time, CR1 and CR2 are not equal, The above CH1 is a heavy chain constant region, The above CL is a light chain constant region, The above L7 and L8 are each independently a peptide linker, wherein L7 is a cleavable or non-cleavable linker, The above f, g, h and z are each independently 0 or 1, and wherein g and h are the same.

26. In paragraph 24 or 25, A fusion protein, wherein the fusion protein further comprises an anticancer agent.

27. In paragraph 26, A fusion protein wherein the above anticancer agent is linked to a fusion protein via a linker.

28. In paragraph 27, A fusion protein, wherein the linker is a non-cleavable linker or a cleavable linker.

29. In paragraph 1, A fusion protein, wherein the fusion protein further comprises an Fc region or a variant thereof.

30. In paragraph 29, The above fusion protein comprises the following structural formulas (I') and (II): N'-T1'-((L1)a-CR1)b-(L2')-I1'-((L3)c-CR3)d-(L5)-Fc region or a variant thereof-C' (I'); and N'-T1''-((L1)a-CR2)b-(L2'')-I1''-((L3)c-CR4)dC' (II) At this time, in the structural formulas (I') and (II), The above N' is the N-terminus of the fusion protein, The above C' is the C-terminus of the fusion protein, The above T1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the first target, The above T1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the first target, The above T1' and T1'' combine to form a binding site (T1) that specifically binds to the first target, The above I1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target, The above I1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target, The above I1' and I1'' combine to form a binding site (I1) that specifically binds to the second target, The above CR1, CR2, CR3 and CR4 are each independently CH1 or CL, At this time, CR1 and CR2 are not equal, CR3 and CR4 are not equal, The above CH1 is a heavy chain constant region, The above CL is a light chain constant region, The above L1, L2', L2'', L3 and L5 are peptide linkers, and L2' and L2'' are cleavable or non-cleavable linkers. The above a, b, c and d are each independently 0 or 1.

31. In paragraph 29, The above fusion protein comprises the following structural formulas (IV') and (V): N'-T2-(L7)-I2'-((L8)f-CR1)g-(L9)-Fc region or a variant thereof-C' (IV'); N'-I2''-((L8)f-CR2)hC' (V) At this time, in the structural formulas (IV') and (V), The above N' is the N-terminus of the fusion protein, The above C' is the C-terminus of the fusion protein, The above T2 is a peptide or antibody fragment that specifically binds to the first target, The above I2' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target, The above I2'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target, The above I2' and I2'' combine to form a binding site (I2) that specifically binds to the second target, The above CR1 and CR2 are each independently CH1 or CL, At this time, CR1 and CR2 are not equal, The above CH1 is a heavy chain constant region, The above CL is a light chain constant region, The above L7, L8 and L9 are each independently a peptide linker, wherein L7 is a cleavable or non-cleavable linker, The above f, g and h are each independently 0 or 1, and wherein g and h are the same.

32. In paragraph 30 or 31, A fusion protein, wherein the fusion protein further comprises an antigen binding site.

33. In paragraph 32, A fusion protein wherein the antigen binding site is linked to the fusion protein via a linker.

34. In paragraph 32, The above fusion protein comprises the following structural formulas (I'') and (II): N'-T1'-((L1)a-CR1)b-(L2')-I1'-((L3)c-CR3)d-(L5)-Fc region or a variant thereof-(L6)eOC' (I''); and N'-T1''-((L1)a-CR2)b-(L2'')-I1''-((L3)c-CR4)dC' (II) At this time, in the structural formulas (I'') and (II), The above N' is the N-terminus of the fusion protein, The above C' is the C-terminus of the fusion protein, The above T1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the first target, The above T1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the first target, The above T1' and T1'' combine to form a binding site (T1) that specifically binds to the first target, The above I1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target, The above I1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target, The above I1' and I1'' combine to form a binding site (I1) that specifically binds to the second target, The above CR1, CR2, CR3 and CR4 are each independently CH1 or CL, At this time, CR1 and CR2 are not equal, CR3 and CR4 are not equal, The above CH1 is a heavy chain constant region, The above CL is a light chain constant region, The above O is an antigen binding site, The above L1, L2', L2'', L3, L5 and L6 are each independently a peptide linker, wherein L2' and L2'' are cleavable or non-cleavable linkers, The above a, b, c, d and e are each independently 0 or 1.

35. In paragraph 32, The above fusion protein is a fusion protein comprising the following structural formulas (IV'') and (V). N'-T2-(L7)-I2'-((L8)f-CR1)g-(L9)-Fc region or a variant thereof-(L10)iPC' (IV''); N'-I2''-((L8)f-CR2)hC' (V) At this time, in the structural formulas (IV'') and (V), The above N' is the N-terminus of the fusion protein, The above C' is the C-terminus of the fusion protein, The above T2 is a peptide or antibody fragment that specifically binds to the first target, The above I2' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target, The above I2'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target, The above I2' and I2'' combine to form a binding site (I2) that specifically binds to the second target, The above CR1 and CR2 are each independently CH1 or CL, At this time, CR1 and CR2 are not equal, The above CH1 is a heavy chain constant region, The above CL is a light chain constant region, The above P is an antigen binding site, The above L7, L8, L9 and L10 are each independently a peptide linker, wherein L7 is a cleavable or non-cleavable linker, The above f, g, h and i are each independently 0 or 1.

36. A fusion protein dimer comprising two fusion proteins selected from the group consisting of items 30, 31, 34 and 35.

37. In paragraph 36, A fusion protein dimer, wherein the above fusion protein dimer is a homodimer or a heterodimer.

38. A fusion protein dimer comprising a fusion protein selected from the group consisting of items 30, 31, 34, and 35; and a fusion protein comprising the following structural formulae (VI) and (VII): N'-I3'-(L11)k-CR1-(L12)-Fc region or a variant thereof-((L13)lV)mC' (VI); and N'-I3''-(L11)k-CR2-C' (VII) At this time, in the structural formulas (VI) and (VII), The above N' is the N-terminus of the fusion protein, The above C' is the C-terminus of the fusion protein, The above I3' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target, The above I3'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target, The above I3' and I3'' form a binding site (I3) that specifically binds to the second target, The above CR1 and CR2 are each independently CH1 or CL, At this time, CR1 and CR2 are not equal, The above CH1 is a heavy chain constant region, The above CL is a light chain constant region, The above V is an antigen binding site, The above L11, L12 and L13 are each independently a peptide linker, The above k, l and m are each independently 0 or 1.

39. A fusion protein dimer comprising a fusion protein selected from the group consisting of items 30, 31, 34, and 35; and a fusion protein comprising the following structural formula (VIII): N'-I3'''-(L14)-Fc region or its variant-((L15)nW)oC' (VIII) At this time, in the above structural formula (VIII), The above N' is the N-terminus of the fusion protein, The above C' is the C-terminus of the fusion protein, The above I3''' is an antibody fragment that specifically binds to the second target, The above W is an antigen binding site, The above L14 and L15 are each independently a peptide linker, The above n and o are each independently 0 or 1.

40. In paragraph 1, The above fusion protein is a fusion protein in which the Fc region of the fusion protein of claim 30 or 31 or a variant thereof is substituted with an antigen binding site.

41. In paragraph 40, A fusion protein wherein the antigen binding site is a heavy chain variable region or a light chain variable region of an antibody.

42. In paragraph 41, The above fusion protein comprises the following structural formulas (I''') and (II): N'-T1'-((L1)a-CR1)b-(L2')-I1'-((L3)c-CR3)d-(L5)-(L18)s-Q'-C' (I'''); and N'-T1''-((L1)a-CR2)b-(L2'')-I1''-((L3)c-CR4)dC' (II) At this time, in the structural formulas (I''') and (II), The above N' is the N-terminus of the fusion protein, The above C' is the C-terminus of the fusion protein, The above T1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the first target, The above T1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the first target, The above T1' and T1'' combine to form a binding site (T1) that specifically binds to the first target, The above I1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target, The above I1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target, The above I1' and I1'' combine to form a binding site (I1) that specifically binds to the second target, The above CR1, CR2, CR3 and CR4 are each independently CH1 or CL, At this time, CR1 and CR2 are not equal, CR3 and CR4 are not equal, The above CH1 is a heavy chain constant region, The above CL is a light chain constant region, The above Q' is a heavy chain variable region (VH) of an antibody that specifically binds to the third target, The above L1, L2', L2'', L3, L5 and L18 are each independently a peptide linker, wherein L2' and L2'' are cleavable or non-cleavable linkers, The above a, b, c, d and s are each independently 0 or 1, and b and d are the same.

43. In paragraph 41, The above fusion protein comprises the following structural formulas (I'''') and (II): N'-T1'-((L1)a-CR1)b-(L2')-I1'-((L3)c-CR3)d-(L5)-(L18)s-Q''-C' (I''''); and N'-T1''-((L1)a-CR2)b-(L2'')-I1''-((L3)c-CR4)dC' (II) At this time, in the structural formulas (I'''') and (II), The above N' is the N-terminus of the fusion protein, The above C' is the C-terminus of the fusion protein, The above T1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the first target, The above T1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the first target, The above T1' and T1'' combine to form a binding site (T1) that specifically binds to the first target, The above I1' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target, The above I1'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target, The above I1' and I1'' combine to form a binding site (I1) that specifically binds to the second target, The above CR1, CR2, CR3 and CR4 are each independently CH1 or CL, At this time, CR1 and CR2 are not equal, CR3 and CR4 are not equal, The above CH1 is a heavy chain constant region, The above CL is a light chain constant region, The above Q'' is the light chain variable region (VL) of an antibody that specifically binds to my target, The above L1, L2', L2'', L3, L5 and L18 are each independently a peptide linker, wherein L2' and L2'' are cleavable or non-cleavable linkers, The above a, b, c, d and s are each independently 0 or 1, and b and d are the same.

44. In paragraph 42 or 43, A fusion protein wherein Q' of the above 42nd clause and Q'' of the above 43rd clause combine to form an antigen binding site (Q) of an antibody that binds to a third target.

45. In paragraph 44, A fusion protein wherein the third target is a protein that is overexpressed or specifically expressed or activated in target cells at the site of the disease.

46. In paragraph 41, The above fusion protein comprises the following structural formulas (IV''') and (V): N'-T2-(L7)-I2'-((L8)f-CR1)g-(L9)-(L19)t-S'-C' (IV'''); N'-I2''-((L8)f-CR2)hC' (V) At this time, in the structural formulas (IV''') and (V), The above N' is the N-terminus of the fusion protein, The above C' is the C-terminus of the fusion protein, The above T2 is a peptide or antibody fragment that specifically binds to the first target, The above I2' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target, The above I2'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target, The above I2' and I2'' combine to form a binding site (I2) that specifically binds to the second target, The above CR1 and CR2 are each independently CH1 or CL, At this time, CR1 and CR2 are not equal, The above CH1 is a heavy chain constant region, The above CL is a light chain constant region, The above S' is a heavy chain variable region (VH) of an antibody that specifically binds to the fourth target, The above L7, L8, L9 and L19 are each independently a peptide linker, wherein L7 is a cleavable or non-cleavable linker, The above f, g, h and t are each independently 0 or 1, and wherein g and h are the same.

47. In paragraph 41, The above fusion protein comprises the following structural formulas (IV'''') and (V): N'-T2-(L7)-I2'-((L8)f-CR1)g-(L10)-S''-C' (IV''''); N'-I2''-((L8)f-CR2)hC' (V) At this time, in the structural formulas (IV'''') and (V), The above N' is the N-terminus of the fusion protein, The above C' is the C-terminus of the fusion protein, The above T2 is a peptide or antibody fragment that specifically binds to the first target, The above I2' is a heavy chain variable region (VH) or light chain variable region (VL) that specifically binds to the second target, The above I2'' is a light chain variable region (VL) or heavy chain variable region (VH) that specifically binds to the second target, The above I2' and I2'' combine to form a binding site (I2) that specifically binds to the second target, The above CR1 and CR2 are each independently CH1 or CL, At this time, CR1 and CR2 are not equal, The above CH1 is a heavy chain constant region, The above CL is a light chain constant region, The above S'' is a light chain variable region (VL) of an antibody that specifically binds to the fourth target, The above L7, L8, L10 and L19 are each independently a peptide linker, wherein L7 is a cleavable or non-cleavable linker, The above f, g, h and t are each independently 0 or 1, and wherein g and h are the same.

48. In paragraph 46 or 47, A fusion protein wherein S' of the above 46th clause and S'' of the above 47th clause combine to form an antigen binding site (S) that binds to a fourth target.

49. In paragraph 48, A fusion protein wherein the fourth target is a protein that is overexpressed or specifically expressed or activated in target cells at the site of the disease.

50. A fusion protein dimer comprising the fusion protein of claim 42 or claim 46; and the fusion protein of claim 43 or claim 47.

51. A polynucleotide encoding a fusion protein according to any one of claims 1 to 35 and claims 40 to 49.

52. A polynucleotide encoding a fusion protein dimer according to any one of claims 36 to 39 and claim 50.

53. A vector comprising the polynucleotide of claim 51 or 52.

54. A transformed cell into which the vector of Article 53 has been introduced. 55.i) A step of culturing the transformed cell of item 54; and ii) A method for producing a fusion protein or a fusion protein dimer, comprising a step of obtaining a fusion protein or a fusion protein dimer from the cell culture medium.

56. A fusion protein-drug conjugate comprising the following structural formula (IX): X-[(L16)-Y]p (IX) At this time, in the above structural formula (IX), The above X is a fusion protein of any one of claims 24, 25, 30, 31, 34, 35, and 42 to 49, The above Y is a drug, The above L16 is a linker, The above p is 0 or 1.

57. In paragraph 56, The above drug is an anticancer drug, a fusion protein-drug conjugate.

58. A fusion protein dimer-drug conjugate comprising the following structural formula (IX'): X'-[(L17)-Z]q (IX') At this time, in the above structural formula (IX'), The above X' is a fusion protein dimer of any one of claims 36 to 39 and claim 50, The above Z is a drug, The above L17 is a linker, The above q is 0 or 1.

59. In paragraph 58, The above drug is an anticancer drug, a fusion protein dimer-drug conjugate.

60. A pharmaceutical composition for the prevention or treatment of a disease, comprising as an active ingredient the fusion protein of claim 1; the fusion protein dimer of any one of claims 36 to 39 and claim 50; the fusion protein-drug conjugate of claim 56; or the fusion protein dimer-drug conjugate of claim 58.

61. In paragraph 60, A pharmaceutical composition wherein the disease is cancer or a degenerative brain disease.

62. In paragraph 61, A pharmaceutical composition, wherein the cancer is any one selected from the group consisting of stomach cancer, liver cancer, lung cancer, colon cancer, breast cancer, prostate cancer, gallbladder cancer, bladder cancer, kidney cancer, esophageal cancer, skin cancer, rectal cancer, osteosarcoma, multiple myeloma, glioma, ovarian cancer, pancreatic cancer, cervical cancer, endometrial cancer, thyroid cancer, laryngeal cancer, testicular cancer, mesothelioma, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, brain tumor, neuroblastoma, retinoblastoma, head and neck cancer, salivary gland cancer, and lymphoma.

63. In paragraph 61, The above degenerative brain disease is any one selected from the group consisting of Alzheimer's disease, Parkinson's disease, Huntington's disease, Mild cognitive impairment, cerebral amyloid angiopathy, Down syndrome, amyloid stroke, systemic amyloid disease, Dutch amyloidosis, Niemann-Pick disease, senile dementia, amyotrophic lateral sclerosis, spinocerebellar atrophy, Tourette's syndrome, Friedrich's ataxia, Machado-joseph's disease, Lewy body dementia, Dystonia, Progressive supranuclear palsy, and frontotemporal dementia. A pharmaceutical composition that is one.

64. A cleavable peptide comprising any one amino acid sequence selected from the group consisting of SEQ ID NOs: 159, 160, 173 to 176 and combinations thereof.

65. In paragraph 64, A cleavable peptide characterized in that the cleavable peptide is cleaved by a protein degrading enzyme that is specifically overexpressed or overactivated in a diseased site compared to a non-diseased site.

66. In paragraph 65, A cleavable peptide, wherein the cleavable peptide is cleaved by any one protease selected from the group consisting of MMP-2, MMP-9, uPA, MT-SP1, MMP-3, MMP-7, and combinations thereof.

67. Use of the fusion protein of claim 1; the fusion protein dimer of any one of claims 36 to 39 and 50; the fusion protein-drug conjugate of claim 56; or the fusion protein dimer-drug conjugate of claim 58 for the prevention or treatment of a disease.

68. A method for preventing or treating a disease, comprising administering to a subject the fusion protein of claim 1; the fusion protein dimer of any one of claims 36 to 39 and 50; the fusion protein-drug conjugate of claim 56; or the fusion protein dimer-drug conjugate of claim 58.

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